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[Effect of passive immunity on subsequent vaccination against swine fever].

Studied was the effect of a serum paratyph and Aujeszky gammaglobulin injected 10, 15, and 20 days prior to the crystal-violet and the lapinized vaccine. If the animals were infected with a pathogenic swine fever virus 10 to 15 days after their treatment with serum at the rate of 0.5 cu. cm per kg, part of them died, and the remaining contracted the disease showing a protracted course. The control vaccination on the 20th day after the animals were injected with serum led to the death of all of them showing the signs of acute swine fever. Most of the pigs injected with 5 cu. cm gamma-globulin each and 10--15 days later being infected with a swine fever virus manifested the disease, however, none of them died of the disease. If, however, after treatment with gamma-globulin they were challenged on the 20th day all of them died of acute swine fever. In the immunization of pigs with a crystal-violet vaccine on the 14th day of their treatment with Aujeszky gamma-globulin, part of them developed an atypical form of the disease, while most of them did not show symptoms whatever of swine fever. Of the immunized with a lapinized vaccine some died, and the remaining part did not contract the disease at all. All this pointed to the fact that the residual antibodies against swine fever contained in the specific Aujeszky gamma-globulin could still be in a position to inhibit the vaccinal virus, and more specifically that of the lapinized K vaccine. The vaccination against swine fever should be performed 14 days after the injection of the serum paratyph and about 20 days after the injection of the Aujeszky gammaglobulin.

[Spread of viral hepatitis in organized children's collectives and the methods for its early diagnosis].

The authors present the results of observations over 407 children aged from 2 months to 16 years from the foci of viral hepatitis in children's collective bodies. During the quarantine a determination was made in children of the glutamic-pyroracemic, glutamic-oxalic transaminases (GPT and GOT, respectively) and of the hepatitis B antigen (HBAg). A necessity of using the enzymatic tests for the purpose of early diagnosis of viral hepatitis was shown, since 84% of the cases developing in the next focus coursed as an unicteric form without any markked clinical signs; HBAg was revealed in 6.1% of the children examined. A complex examination of the personnel and of the persons who came in contact with the patients with viral hepatitis showed the ways of spread of hepatitis B in a collective body; it was found that the viral hepatitis B infection took place both by parenteral and enteral routes. The expediency of active observation over the children, recipients of blood and plasma, with determination in them of the activity of the enzymes and HBAg for early diagnosis of parenteral infection was substantiated. It was also shown that the incidence of the unicteric forms of viral hepatitis in a focus of infection depended not on the periods of gamma-globulin administration but on the age of children who contracted the infection. Thus, the prevalence of the unicteric forms of the disease over the icteric ones in children under 3 years of age was more pronounced than in older children.

A controlled study of allergen production in cultures of Dermatophadoides pteronyssinus.

Allergenic substances have been isolated from cultures of Dermatophagoides pteronyssinus on wheat germ flakes and powdered yeast; as controls, non-sterile nutrient medium containing no mites, as well as sterile medium, were maintained and extracted under identical conditions. Chemical purification and analyses indicated the occurrence of skin-active allergens in all three preparations. This was confirmed by the assay of skin-reactivity in vivo and complement-inactivating capacity in vitro. Quantitatively, the medium containing bacteria and fungi contained more allergen than the sterile control, but less than the mite-infected medium. The evidence indicates that degradation reactions of nutrient components proceed faster in the non-sterile media than in the sterile control. The results of immunochemical analyses demonstrate that degradation reactions, give rise to the introduction of lysine-sugar structures inot high-molecular weight components from the medium. It is concluded that mites and micro-organisms have a role in allergen formation by promoting degradation reactions among nutrient constituents.

Chondroitin sulfate and electron lucent bodies in the pericellular rim about unshrunken hypertrophied chondrocytes of chick long bone.

Direct observation of unstained, 1 mm thick blocks of fresh epiphyseal cartilage from tibia of 15- and 18-day-old chick embryos revealed shrunken chondrocytes on its cut surfaces but unshrunken chondrocytes deep within the tissue blocks. The unshrunken hypertrophied chondrocytes are rimmed with refractile substance identified as chondroitin sulfate removable with hyaluronidase. This substance is stained metachromatically red with toluidine blue, and is stained with ruthenium red and with ruthenium red-OsO4. The latter, observed with the electron microscope, is present as an electron dense rim, specifically about the unshrunken, hypertrophied chondrocytes between the plasma membrane and lacunar wall. By rendering the chondroitin sulfate electron dense with RR-OsO4, electron lucent bodies (ELB) were revealed specifically about the hypertrophied chondrocytes. The ELB contain an electron dense core with radiating fibrils. The content and source of ELB, also found in the intercellular matrix, are not known. The 0.1% toluidine blue solution containing 0.2 M MgC12 or 0.4% NaCl or KCl stained juxtanuclear clusters of granules metachromatically red. The location of intracellular granules was believed to represent a cluster of Golgi-derived vesicles. The pericellular metachromatic, RR-OsO4-positive rim is believed to be an accumulation of externalized juxtanuclear metachromatic granules. The possibility that the ELB may also be externalized content of Golgi vesicles was entertained.

Alpha-Fetoprotein levels in normal males from seven ethnic groups with different hepatocellular carcinoma risks.

Alpha-Fetoprotein (AFP) levels of 1,335 males (15 years and older) of seven ethnic groups (Chinese, Indians, and Malays from Singapore, Caucasians from Lyon, and Blacks from Nairobi, forest, and the savanna region of the Ivory Coast) were determined by radioimmunoassay. A few elevated levels (up to 30 nanounits/ml) were detected in some normal individuals, especially in the older age-groups. In addition, there was a systematic age-dependency of AFP levels particularly evident in the groups from Singapore-Lyon, in which there was a 50% AFP increase between the ages of 20 and 40. Comparison between Africans on the one hand and people from Singapore-Lyon on the other hand revealed highly significant differences (p less than 0.001), especially in the younger groups, whereas Chinese, Malays, and Indians from Singapore had very similar AFP pattern; this suggests an important role for environmental factors in the regulation of AFP levels. The age dependency of the presumed effect of environmental factors is in keeping with experimental data showing that young animals respond more vigorously to AFP-stimulating factors. Although the incidence of hepatocellular carcinoma (HCC) differs in the three Singapore groups (the highest in Chinese and the lowest in Indians), no relationship was observed in this study between mean AFP level and HCC incidence in Singapore.

Alpha-Fetoprotein and albumin synthesis during the cell cycle.

The synthesis of alpha-fetoprotein and albumin in two clones of AH66 hepatoma was studied. (1) Amounts of AFP and albumin synthesized by the C-4 clone were 2.9 and 0.28 X 10(-7) mug per cell per hour, respectively. AFP and albumin amounts synthesized by the A-1 clone were 2.5 and 1.8 X 10(-7) mug per cell per hour, respectively. (2) The cell cycles of the C-4 and A-1 clones were as follows: C-4 clone: mean generation time 20.5 hours: G1, 10 hours; S, 7 hours; G2, 4 hours; and M, 30 minutes. A-1 clone: mean generation time 50.7 hours: G1, 36 hours; S, 10 hours; G2, 4 hours; and M, 30-60 minutes. (3) AFP was found to be synthesized from late G1 phase to the end of the S phase for 9 hours in C-4 and 25 hours in A-1. The albumin production was from late S phase to the beginning of the G2 phase for 4 hours in C-4 and 9 hours in A-1, which were approximately half or one-third of the time spent on AFP production. (4) The double staining with fluorescent-conjugated antibodies to AFP and albumin demonstrated that AFP and albumin are probably synthesized by different cells.

Ionic regulation of the haemolymph in the larvae of the dragonfly Aeshna cyanea (Müller) (Odonata, Anisoptera).

The ionic regulation of the haemolymph of larvae of Aeshna cyanea (Müller) was studied by means of two types of experiments. In the first the change in internal ionic composition was followed as a function of the time spent in a given experimental medium. These experiments led to the conclusion that: 1. the haemolymph composition does not change when larvae are starved in tap water for 10 days; 2. the haemolymph ionic concentrations (Na and Cl) have initial marked increase when the animals are kept in hypertonic media of diluted sea water; after 80 hours however, both concentrations stay constant. In a second series of experiments the internal ionic concentration was compared to a series of different concentrations of external media. From this, the relation between the internal and external ionic concentration was elaborated : in hypotonic media the internal Na and Cl concentrations stay constant, in hypertonic media there is a parallelism between the increase of the external concentration and haemolymph concentration, the internal Na concentration being always slightly hypertonic, the Cl concentration markedly hypotonic. Finally, when larvae are placed in glass distilled water, the internal Na and Cl concentrations begin to decrease after 60 hours.

Interaction between lindane and micorbes in soils.

Three lindane (gamma-1,2,3,4,5,6-hexachlorocyclohexane) treated soils were studied under laboratory conditions to determine the interaction between lindane and the soil microorganisms. Microbial populations and respiration were monitored to study insecticide effects. Formation of lindane degradation products and chloride content were examined to determine effects of the microorganisms. Some populations in lindane treated soils showed temporary declines but all ultimately recovered to at least the level of the controls in 16 weeks. Respiration was stimulated over a 9-week period especially in the sandy and clay loams, suggesting the possibility of microbial degradation of the insecticide. Lindane degradation products separated and identified by TLC included gamma-2,3,4,5,6-pentachloro-1-cyclohexene (gamma-PCCH), gamma-3,4,5,6,-tetrachloro-1-cyclohexene (gamma-TCCH), gamma-3,4,5,6-tetrachloro-1-cyclohexene (gamma-TCCH), and pentachlorobenzene (PCB). Chloride production increased in soils treated with higher levels of lindane.

2-phenylethanol and some of its amphiphilic derivatives as inhibitors of platelet aggregation. Structure-activity relationship.

The relationship between chemical structure and inhibitory activity of some simple water soluble phenylalkyl derivatives has been investigated. The inhibitory effect of the following analogous and homologous derivatives increases in the sequence: phenylacetic acid less than 2-phenylethanol less than 2-phenylethylamine = 4-phenylbutyric acid less than 3-phenylpropanol less than 4-phenylbutylamine, i.e., the inhibitory effect is potentiated. 1. by elongation of the alkyl chain, which is accompanied by an increase of the lipophilic character, and 2. when the polar portion is represented by the positively charged amino-group. The effect of these amphiphilic compounds on platelet aggregation appears to be mediated through polar and apolar interactions with the outer layers of the plasma membrane. The fact that the amino derivatives are the most active inhibitors suggests that negatively charged groups may be involved in the polar binding of these inhibitors. Experiments with neuraminidase-treated platelets revealed that the enzymatically removable sialic acid residues are not those negatively charged binding sites. Phentolamine and propranolol which block adrenergic alpha- and beta-receptors, respectively, do not influence the inhibitory effect of these agents.

Effect of thiouracil upon canine arterial elastic tissue.

Unlike some other mammalian species, the dog is relatively resistant to the development of elevated levels of serum cholesterol after prolonged cholesterol feeding. This may be overcome by suppressing thyroid activity with thiouracil. Information regarding possible activity of thiouracil itself upon the arterial tissues is almost nonexistent. The present investigation was undertaken to test whether this drug has any such action, especially upon the arterial elastic tissues. Destructive changes were observed in arterial elastic tissues in dogs given thiouracil for three and six months. The changes consisted of accentuation of the elastic fibrillar components, formation and subsequent coalescence of clefts, and fragmentation and ultimate "dissolution" of the elastic elements. The results suggest that thiouracil may exert a damaging effect upon the arterial elastic fibers; thus, it is possible that one of the mechanisms by which thiouracil and cholesterol administration induces experimental atherosclerosis in the dog is by elastic tissue destruction, possibly promoting the subsequent lipid accumulation in the arterial wall.

Evidence for significant hematopoiesis in the human thymus.

Earlier studies on fetal thymus suggested that certain of the large pyroninophilic cells found there might have a hemopoietic role, and it was decided to determine the nature of these cells using histochemical and immunohistochemical methods. Thymic tissue from aborted fetuses, stillbirths, and neonatal deaths was examined histochemically using methods for the detection of chloroacetate esterase, peroxidase, and pseudoperoxidase, and by staining techniques for mast cells and eosinophils. Tissue was also examined using the indirect immunoperoxidase method for the presence of hemoglobin A (HbA) and F (HbF), for lysozyme (muramidase) and immunoglobins alpha, mu, gamma, kappa, lambda. Positive staining to some degree was seen in cells in the connective tissue stroma using all methods, and the cells stained corresponded to one or another of the types of pyroninophilic cells present. The finding of large cells with positive chloroacetate esterase and antilysozyme indicates the presence of granulopoiesis. Similarly, the presence of large nucleated cells with pseudoperoxidase and anti-hemoglobin (A and F) staining indicates the presence of erythropoiesis. Plasma cells were present in small numbers.

Studies on the inhibition of macrophage migration induced by soluble antigen-antibody complexes.

Mixtures of serum of Freund's complete adjuvant (FCA) immunized guinea-pigs and tuberculin PPD consistently inhibited the in vitro migration of peritoneal exudate cells (PEC) of normal guinea-pigs. It is shown that this inhibitory effect is due to a soluble complex between an IgG2 antibody and PPD. By separation of PPD on Sephadex G-200 two peaks were obtained, corresponding respectively to molecular weights of at least 800,000 and 25,000, separated by a plateau. Material derived from both peaks and from the plateau was able to form inhibitory complexes with anti-PPD IgG2. When a mixture of small molecular weight PPD and anti-PPD IgG2 was fractionated on Sephadex G-200, the inhibitory activity was recovered in the void region only. The detection of both IgG2 and PPD in the latter was taken as evidence for the presence of a high molecular weight antigen-antibody complex. When the mechanism of complex-induced inhibition of migration was examined it was found that: (1) complexes act directly on macrophages present in the peritoneal exudate; (2) removal of the Fc fragment of IgG2 by pepsin abolishes its ability to form migration inhibitory complexes; (3) passive sensitization of macrophages with anti-PPD IgG2, followed by exposure to PPD does not result in inhibition of migration; (4) in order to obtain migration inhibition, the complexes must be present during the entire migration period. A 2-hr pulse with complexes does not induce permanent inhibition; (5) the migration inhibitory activity of antigen--antibody complexes can be abolished by certain concentrations of puromycin and aminophylline.

Sex-dependent differences in the biosynthesis of pyrimidine nucleotides in rat liver after repeated administration of alpha-hexachlorocyclohexane.

Utilization of [2-14C]orotic acid for the synthesis of the pyrimidine components of the free nucleotide pool and RNA from isolated cytoplasmic ribosomes was investigated in the liver of female and male rats. In females, labeled orotic acid is incorporated relatively more into uridine than into cytidine nucleotides; the opposite is true for males. The administration of alpha-hexachlorocyclohexane (alpha-HCH) decreases the specific radioactivity of rRNA cytidylic acid more in males than in females. Simultaneously, the level of liver microsomal cytochrome P-450 is increased after the administration of alpha-HCH more in males than in females. After four days of treatment with alpha-HCH, the concentrations of the uridine and cytidine components of the acid-soluble pool are slightly depressed. Following repeated administration of alpha-HCH, the utilization of labeled uridine for the synthesis of cytidine nucleotides of the acid-soluble pool and RNA is depressed, whereas that of labeled cytidine is enhanced. Repeated administration of alpha-HCH decreases the utilization of [2-14C]orotic acid for the synthesis of cytidine nucleotides of DNA; the specific radioactivity of thymidylic acid is increased. The administration of beta-diethylaminoethyl diallylacetate (CFT-1201) to animals that have repeatedly received alpha-HCH decreases the specific radioactivity of DNA thymine.

Antigenic structure of the cyanogen bromide peptide F-CB3 from fibrinogen alpha-chain.

The antigenic properties of the cyanogen bromide peptide F-CB3 from bovine fibrinogen alpha-chain were studied in radioimmune assays with rabbit antibodies to fibrinogen or to peptide F-CB3. Both fibrinogen and peptide F-CB3 were indistinguishable in inhibition and dissociation tests. Modification of the single disulfide bridge in peptide F-CB3 either by reduction or by cleavage with cyanide was not accompanied by loss of serologic activity. Inhibition studies with three individual fragments obtained after cyanide cleavage (molecular weight range 7000 to 23000) indicated the presence of at least three distinct antigenic determinants in peptide F-CB3. After trypsin digestion of peptide F-CB3 still 75% of its maximal inhibiting capacity was retained. Lack of change in antigenic activity of peptide F-CB3 after release from the fibrinogen molecule by cyanogen bromide and upon further fragmentation is presumably due to the presence of several sequential antigenic determinants but the presence of conformational determinants could not be entirely excluded. Since no cross-reaction was observed between bovine and human peptides F-B3 one may expect considerable variation in their amino acid sequence.

["Hairy cell" leukemia].

By means of individual observations the clinical, cytological and cytochemical marks of the so-called "hairy cell"leukemia are described. Peculiarities in the course of the disease and in prognosis justify this rare variant to be separated from reticular systems diseases. A diagnosis can also be made on the basis of cytomorphological characteristics.

Molecular construction of Clostridium botulinum type A toxins.

Two Clostridium botulinum type A toxic fractions, named large (L) and medium (M) toxins, were eluted from Sephadex G-200. Sucrose density gradient centrifugation resolved L toxin (2.5 X 10(8) to 3.0 X 10(8) mean lethal doses per mg of N) into two fractions, 19S and 16S. The same procedure performed at pH 8resolved it into three fractions; the heavier two were both nontoxic and hemagglutinin positive, and the lightest on (7S) was toxic. M toxin (12S) (4.5 X 10(8) to 5.0 X 10(8) mean lethal doses per mg of N) was homogeneous in electrophoresis and centrifugation at pH 6. The latter procedure performed at pH 8 dmonstrated that it dissociated into uniform 7S components. The nontoxic component of M toxin was free from hemagglutinin. M toxin alone was demonstrated in culture by sucrose density gradient centrifugation at pH 6. Dialysis of the culture supernatant resulted in partial formation of 16S toxin. Centrifugation of the crystalline toxin in 1 MNaCl demonstrated 16S toxin only. The toxic components of L, M, and crystalline toxins were antigenically identical. The nontoxic components of the crystalline and L toxins, consisting of two distinct antigens, were antigenically identical; that of M toxin was identical with one of these two antigens.

Infection in immunodepressed patients. The approach to diagnosis and treatment.

Infection is an important cause of death in patients receiving cytostatic drugs or with any other impairment of host resistance. Such infections are frequently due to opportunist micro-organisms usually belonging to the endogenous flora of the patient. It is often difficult to obtain an exact diagnosis of the cause and localization of the infection. The problems associated with the prevention of infection are manifold. Exogenous infections can be prevented by proper isolation and a sterile diet. Endogenous infections can only be prevented by eradication of the patient's endogeous flora, so-called decontamination. Special attention should be given to treatment of foci of chronic infection and of the carrier state of certain microorganisms. However, the prophylactic use of antibiotics should be avoided. The curative use of antibiotics should be based on the most probable micro-organism. We consider the inventory of the patient's microflora, repeated weekly, of great help in the choice of antibiotics in cases of septicaemia of unknown aetiology. The initial therapy usually consists of a broad-spectrum combination of antibiotics, which should be bactericidal. When the causative bacteria have been isolated and the sensitivity is known, antibiotic therapy should be adjusted to the narrowest spectrum possible.

An ultrastructural study of acid phosphatase localization in Phaseolus vulgaris xylem by the use of an azo-dye method.

The localization of acid phosphatase during xylem development has been examined in the bean, Phaseolus vulgaris. The azo dye, the final reaction product, is initially prominent in the dictyosomes, vesicles apparently participating in secondary wall formation, and in the middle lamella of the young vessel element. Final reaction particles are also present in mitochondria, chloroplasts, and certain vacuoles and are sparsely scattered in the cytoplasm. At a later stage of vessel differentiation, the azo dye is concentrated in the disintegrating cytoplasm and along the fibrils of the partially hydrolysed primary wall and middle lamella. In the mature vessel element, the azo dye is still present along the disintegrated primary wall at the side of the vessel and covers the secondary wall. In the parenchyma cell adjacent to the vessel element, acid phosphatase localization is found in the dictyosomes, endoplasmic reticulum, mitochondria, small vacuoles, and the middle lamella. The controls from all stages of vessel element development were free of azo dye particles. The concentration of acid phosphatase along the secondary walls of the mature vessels and in the middle lamella between other cells indicates that this enzyme has other functions besides autolysis of the cytoplasm and primary cell wall. Acid phosphatase may participate in the formation of the secondary wall and may also have a role in the secretion and transport of sugars.

Degradation of blood group antigens in human colon ecosystems. II. A gene interaction in man that affects the fecal population density of certain enteric bacteria.

The autosomal dominant ABH secretor gene together with the ABO blood type gene control the presence and specificity of A, B, and H blood group antigens in human gut mucin glycoproteins. Certain obligate anaerobes in feces produce extracellular antigen-specific glycoside structures. We estimated the populations of these bacteria in feces of 22 healthy subjects by determining the greatest dilution of feces that yielded A, B, or H blood group-degrading enzyme activity after 24 h incubation in anaerobic cultures. Comparatively small populations of fecal bacteria produce blood group-degrading enzymes; their estimated populations were 10(8) per g or less in 21 subjects. Fecal populations of B-degrading bacteria were stable over time, and their population density averaged 50,000-fold greater in blood group B secretros than in other subjects. We present evidence that the greater fecal populations of B-degrading bacteria in B secretors is due in part to a competitive nutritional advantage gained by their ability to enzymatically cleave the B antigenic determinant alpha-D-galactose from gut mucins of B secretors. Fecal populations of bacteria producing A and H antigen-degrading enzyme activities were comparable in all subjects to the fecal population of B-degrading bacteria in B secretors. The large populations of fecal anaerobes may be an additional source of A antigen substrate for A-degrading bacteria; thus, antigens cross-reacting with A antigen were detected on cell walls of anaerobic bacteria from 3 of 10 cultures inoculated with 10(-10) g feces. Bacteria producing B-degrading activity likely represent a separate population from those producing A- or H-degrading activity since their fecal populations differed numerically in 14 subjects. These findings suggest that adaptation of blood group-degrading enzymes to mucin structures in human colon ecosystems is chiefly by mutation-selection of comparatively small populations of constitutive enzyme-producing strains rather than by substrate induced enzyme synthesis in many strains.

Physicochemical characterization of direct fluorescent antibody reagents.

When the data from performance and physicochemical studies of conjugates are combined for analysis, the performance data and specific titers show a direct relationship to the physicochemical data (Table 2). These reagents were prepared from the same lot of antiserum. The specific titers are very misleading without the accompanying data (Table 2). The protein concentrations range from 4 to 10 mg/ml, the F/P ratios from 10 to 30, and CASE shows gamma-globulin to constitute 30 to 100% of the protein. CASE also shows the gamma-globulin F/P ratio to be only 10 to 20. Using these data, we calculated the concentrations of the gamma-globulins and normalized their titers to 10 mg/ml. The value of good fractionation procedures for recovering gamma-globulin and the desirability of obtaining optimal F/P ratios are reflected in the adjusted titers. Physicochemical characterization of conjugates identifies superior and deficient reagents and frequently reveals the cause of inadequate performance. In this way it serves as a quide for improving reagent quality.

Suppression and reversal of allergic encephalomyelitis in guinea pigs with a non-encephalitogenic analogue of the tryptophan region of the myelin basic protein.

The administration of synthetic peptide S42 leads to suppression and reversal of experimental allergic encephalomyelitis (EAE) induced in guinea pigs by myelin basic protein. Peptide S42 contains a linear sequence of 21 amino acid residues, H-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Gly-OH, made up of four repeating unit sequences of H-Phe-Ser-Trp-Gln-Lys-OH in addition to a C-terminal glycine. Injected at relatively high doses, peptide S42 is non-encephalitogenic. It induces delayed-type hypersensitivity which is not followed by EAE, and elicits delayed-type hypersensitivity responses in peptide S42, encephalitogenic trytophan peptide, or BP-challenged animals for either of the three antigens. The repeating unit sequence of peptide S42 is analogous to the encephalitogenic tryptophan region of the BP molecules . The sequence homology is responsible for cellular recognition of this antigen by the skin test assay and suggests in vivo interaction between peptide S42 and EAE-inducing cells leading to suppression and reversal of disease.

Antigenic determinants of adenovirus capsids. II. Homogeneity of hexons, and accessibility of their determinants, in the virion.

We have tested the two principal theories which explain the previous finding that small amounts of type-specific antibody to the adenovirus hexon can neutralize infectivity, whereas even large amounts of cross-reactive antibody do not. a) It has been suggested that the type-specific determinants are especially prominent in the virion. We have therefore measured the capacity of whole virus to bind appropriate antibodies, using a sensitive radioimmunoprecipitation (RIP) system. In fact, virions bound type-specific and cross-reactive antibodies impartially. Moreover, they bound both much less effectively than did free hexon or disrupted virus, suggesting that many of each kind of determinant are inaccessible in virions. b) It has been suggested that the type-specific determinants are confined to those hexons located next to the pentons, and that they are the targets for neutralizing antibody. We have therefore studied the antigenicity of peripentonal and nonamer hexons isolated from virions, and found that each possessed both kinds of determinants. Furthermore, these were present in the same proportion as in hexons purified from the soluble antigens in infected cells ("free hexons"). We concluded that the mechanism of neutralization by antibody is complicated, and that the type-specific determinants exposed on the virion must play a crucial role.

Evaluation, surveillance and treatment of panoral leukoplakia.

Leukoplakia is well-recognised as a premalignant condition of the oral mucosa. Despite many attempts to produce definitive laboratory tests for the prediction of those cases of premalignant leukoplakia which may undergo malignant transformation, there is as yet no fool-proof method of clinical and laboratory assessment. The problem is nowhere more manifest than in the management of panoral leukoplakia, especially in frail, elderly and apprehensive patients. In such cases the selection of sites for biopsy and treatment is often more empirical than logical. The introduction of cryosurgical techniques for the treatment of leukoplakia has enabled the surgeon to eradicate panoral white patches in a more conservative fashion. The application of a diagnostic nuclear staining test using toluidine blue dye, coupled with local exfoliative cytology, has provided a system for evaluation and surveillance of panoral leukoplakia which is clinically simple and at least as reliable as any predictive test previously described.

Alpha1-antitrypsin in amniotic fluid and cord blood of preterm infants with the respiratory distress syndrome.

Total protein, alpha1-antitrypsin, and alpha2-macroglobulin were measured on amniotic fluid in 125 pregnancies between 11 and 42 weeks' gestation, and on the cord blood of 66 newborn infants. Amniotic fluid surface active material was assessed by the foam stability test. Amniotic fluid alpha1-antitrypsin is linearly and directly related to amniotic fluid total protein (r = 0.703, p less than 0.001). The cord alpha1-antitrypsin is also linearly related to cord total protein. Intrapartum complications are associated with a significant lowering of the cord alpha1-antitrypsin. Infants with a negative foam stability test had RDS regardless of the amniotic fluid alpha1-antitrypsin. The cord blood alpha1-antitrypsin value did not appear to be related to amniotic fluid surface active material. There were 23 infants with cord alpha1-antitrypsin of less than 0.2 gm/dl and with an intermediate or positive foam stability test; 19 of them had respiratory difficulties of varying severity. It is conceivable that infants, in spite of apparent adequate prenatal lung surfactant, develop respiratory disturbances on the basis of pulmonary fluid and protein transudation and/or reduction or inhibition of pulmonary surfactant incident to intrapartum complications.

Effect of long- and short-term intravaginal progestagen treatments on synchronization of oestrus and fertility in heifers.

A total of 273 Herford cross heifers were treated with intravaginal progestagen pessaries to determine some of the factors affecting oestrous response and fertility following long-term (20-day) and short-term (10-day) treatments. Oestrous response and degree of synchronization were high after treatment for 20 days, but the fertility rate was lower than that of control heifers. There was no difference in the fertility of heifers inseminated artificially and those mated naturally. When the treatment period was reduced to 10 days and 900 mg progesterone and 5 mg oestradiol valerate given intramuscularly at the start, a high oestrous response and a low degree of synchronization resulted, but the conception rate was similar to that of the control animals. Reducing the dose of progesterone to 250 mg resulted in a high oestrous response and a high degree of synchronization. The stage of the cycle at the start of the 10-day treatment did not affect the oestrous response. Retention of the progesterone pessary was low (79-9%) in heifers treated for 2- days, but was 100% in those treated for 10 days.

Factors determining the susceptibility of mice to experimental phycomycosis.

Various factors influencing the susceptibility of C3H mice to lethal infection by the phycomycete Absidia ramosa were examined. Mice 19-21 days old were exposed to graded doses of A. ramosa spores by various routes. After intravenous inoculation with doses in excess of 10(3) viable units, a variable proportion of mice developed lethal phycomycosis of the central nervous system within 2-8 days. The proportion of mice affected was related to the inoculum size, doses of 5 X 10(7) spores producing lethal infection in 90-100% of the mice. The disease was characterised by signs of involvement of the central nervous system appearing 8-12 h before death. At necropsy, fungal hyphae, frequently surrounded by infiltrations of mononuclear cells, could be demonstrated in the brain. Lesions were also often present in the kidneys; in other organs they were rare, but the presence of viable fungal spores could be detected by cultural procedures. Subcutaneous inoculation of A. ramosa spores did not produce lethal infection but intraperitoneal inoculation occasionally did so. Direct intracerebral inoculation invariably produced lethal infection even with very small doses of spores. The clinical and histopathological features of the disease were almost identical with those resulting from intravenous inoculation. Pre-inoculation treatment with azathioprine, cyclophosphamide and antithymocyte serum did not increase susceptibility. Susceptibility was increased by injections of zymosan, aggregared gamma-globulin, cortisone acetate and Fe(II) salts and by reticuloendothelial blockade. These treatments increased the proportion of mice developing lethal phycomycosis of the central nervous system, and in the case of cortisone acetate, also promoted disseminated infection. It was concluded that the natural resistance of mice to A. ramosa infection was probably dependent upon the activity of phagocytic cells, possibly acting in conjunction with complement and other non-specific serum factors.

[Ventricular fibrillation in acute phase of myocardial infarct. 1. Relationship between the development of ventricular fibrillation in myocardial infarct and previous rhythm disorders].

The author analyses the results of experimental studies in dogs and cats, that included a continuous recording of ECG, electrogram and monophase cardiac potentials during 1 hour following coronary artery ligation. The ligation caused bradycardia, but no correlation was found between the degree of bradycardia and the development of extrasystole and ventricular fibrillation. Extrasystole developed in 100% of the experiments in which the coronary artery ligation resulted in ventricular fibrillation, and in 66% and 87% of those without this complication, conducted in dogs and cats respectively. The rate of extrasystole proved important for the prognosis of fibrillation. The number of extrasystoli noted during the mean time of the development of fibrillation was 5 times higher in the experiments with ventricular fibrillation than in those without fibrillation. In the experiments with fibrillation the extrasystoli tended to occur earlier within the cardiac cycle. Of the total number of extrasystoli, grouped extrasystoli comprised 89% in the experiments with ventricular fibrillation, and 21%--in those without fibrillation. Ventricular tachystystole was noted in 50% of the experiments with fibrillation and in 17% of those without this complication. In the experiments complicated by fibrillation the period of ventricular tachysystole was characterized by a gradual shortening of the cardiac cycles.

Effect of low-dose heparin prophylaxis on arterial oxygen tension after high laparotomy.

The effect of low doses of heparin (5000 units of sodium heparin every 12 hours for 5 days) on arterial oxygenation was studied in 24 patients in the postoperative period after upper abdominal surgery. Another 24 patients served as a control group. The arterial oxygen tension was the same in both groups preoperatively and was equally significantly reduced during the 1st postoperative day. During the 2nd day, oxygen tension rose in the heparin-treated group to values which no longer differed significantly from the peroperative level. In the control group the significant reduction persisted until the 4th postoperative day. The arterial carbon-dioxide tension did not differ between the groups, neither did it vary significantly between days. There were no clinical signs of large pulmonary embolism during the postoperative period, chest X-ray was normal in all patients examined and a photoscan was normal in 23 of 24 subjects studied. Low-dose heparin treatment may apparently shorten the period of postoperative hypoxaemia, probably by counteracting both large pulmonary emboli and microthromboembolism.

IgA deficiency and severe post-vagotomy diarrhoea.

Between January, 1969, and December, 1973, 2058 truncal vagotomies were performed in the Merseyside Regional Health Authority area. 14 of these patients subsequently developed severe post-vagotomy diarrhoea and were extensively investigated. 6 were found to have IgA deficiency. It is suggested that antecedent IgA deficiency may account for the varied reported incidence of severe post-vagotomy diarrhoea and that preoperative screening could reduce the incidence of this complication.

Sodium excretion and sympathetic activity in relation to severity of hypertensive disease.

The relationship between the severity of hypertensive disease and sodium excretion and sympathetic activity has been studied in subjects of the same age and sex derived from screening a total population. 19 untreated subjects with casual blood-pressure (B.P.) above 175/115 mm Hg on two separate occasions made up the hypertensive group. A normotensive group (n =19) was obtained by selecting a 5% random sample from all subjects with casual B.P. below 160/95. Sympathetic activity was determined from noradrenaline excretion and the severity of hypertension assessed by recording resting diastolic B.P., signs of left ventricular hypertrophy on orthogonal E.C.G., and the glomerular filtration-rate. In the hypertensive group the resting B.P. correlated well both with signs of left ventricular hypertrophy and with the glomerular filtration-rate--i.e., the degree of severity of the hypertension. Up to the level of 90 mm Hg resting diastolic B.P., sodium excretion rose in complete agreement with the theory of pressure diuresis. Above 90 mm Hg, however, sodium and noradrenaline excretion fell with increase of B.P. These two findings indicated that with increasing severity of hypertension the sodium balance overrides the sympathetic activity in the long-term regulation of B.P. This may have both prognostic and therapeutic implications.

[The role of chlorazepate dipotassium (Tranxilium) in the therapy of psychosomatic syndromes (author's transl)].

Divided into four groups according to different kind and cause of disorder, 240 patients showing psychosomatic disorders have been treated with chlorazepate dipotassium only or in combination with clomipramine and dihydroergotamine tartrate ambulant or in hospital, depending on the degree of severity of the disorder. With 101 clinically treated cases of cyclothymic depression good results were obtained with combined treatment with chlorazepate dipotassium while reducing the dose of the antidepressant. The same result was obtained with 63 patients suffering from severe neurasthenic exhaustion and 13 patients with general neurodystonic symptoms treated with chlorazepate dipotassium only. The combination of the usual dihydroergotamine tartrate medication with a chlorazepate dipotassium treatment over several months showed longlasting good therapeutic results, confirmed by follow-up examinations, in 31 out of 40 cases with migraine respectively vasomotor headache. In the other nine patients with migraine the complaints persisted only in rare instances.

[Cytochemical demonstration and characterization of acid mucopolysaccharides in normal granulocytic series].

Metachromatic granulations are seen in myeloblasts, promyelocytes, myelocytes, metamyelocytes and young mature cells, after staining by all metachromatic dyes, except methylene blue. Furthermore by screening at various pHs granular metachromasia is detectable starting at pH 3.2 in 6% of case, at pH 4 in 78% of cases, at pH 4.5 in 16% of cases. The use of toluidine blue 10(-4) M in solutions at varying pH, after methylation and after alkali demethylation demonstrate the acid sulfated mucopolysaccharide react within the pH range (3.2), 4, 4.5, 5; persistence of non-alcohol resistant granular metachromasia above pH 5.5 suggests the presence of granules containing acid mucopolysaccharides staining as the result of possessing --COOH radicals. This data was verified by other histochemical techniques (Alcian blue and PAS staining) which have shown positive granules. All the data on the metachromatic, alcian blue and PAS positive granulations, have been carefully confirmed by chemical and enzymatic reactions. The results of this study led us to the conclusion that azurophilic granulations are made of a sulfated polysaccharide of chondroitinsulfate group. In the more immature cells alcian blue positive granules (mucopolysaccharide of acid hyaluronic type) and PAS positive granules (polysaccharides with a low sulfation level) can be demonstrated. From this data, it is suggested that azurophilic granulations are the final event of an evolutive process of polysaccharide material, fundamentally based on a progressive sulfation.

A molecular concept of the properdin pathway.

The sequential events of the properdin system were analyzed. Properdin-depleted serum allows the formation of a Factor B- and D-dependent C3 convertase. This enzyme, called the properdin-receptor-forming enzyme, was shown to utilize a novel serum component, the initiating factor. The protein is a beta-globulin in precursor form and is distinct from immunoglobulins. The function of the enzyme is to deposit C3b on the surface of activator particles. Apparently doublets of C3b are required for the formation of the properdin-activating principle. It consists of a complex containing surface-bound C3b and activated Factor B. properdin precursor is activated by binding to this complex without detectable change in molecular weight. The transition of properdin precursor to activated properdin is probably caused by a conformational change. The complex, consisting of bound C3b, properdin, and activated Factor B, represents the enzyme that acts on C5, thereby initiating self-assembly of the membrane attack system. Native C3 is not needed for the function of the enzyme. It is disassembled by soluble C3 or C3b and its formation is under the control of the properdin-receptor-destroying enzyme, which may be identical with the C3b inactivator.

Radionuclide joint imaging.

Modern radionuclide techniques of joint imaging involve the use of either 99mTc-pertechnetate or 99mTc-phosphate compounds in conjunction with the Anger camera. In general, images obtained with both types of radiocompound are nonspecific--although increased uptake of 99mTc-pertechnetate usually denotes the presence of synovitis. The most popular uses of the technique are in documenting the extent and severity of inflammatory joint disease, assessing the effect of therapy, and establishing the diagnoses of Legg-Perthes disease and septic arthritis. The method is also useful in judging the extent of involvement in osteoarthritis of the knee prior to surgical intervention. Radionuclide joint imaging is more sensitive than clinical or radiographic techniques in detecting early joint involvement but usually it must be supplemented by other techniques to establish a specific diagnosis.

The effect of bleomycin on rapidly proliferating epidermis. A comparative investigation using micro-flow fluorometry, H3Tdr incorporation and a stathmokinetic method (colcemid).

At different time intervals after injection of Bleomycin (BLM) th effect on several kinetic parameters of the hairless mouse epidermis stimulated to proliferate by previous adhesive tape stripping was measured. Micro-flow fluorometry was used to determine the relative number of cells in the various phases of the cell cycle (G1, S and G2). Tritiated thymidine was used to determine labelling indices and grain counts. Colcemid was used to observe the mitotic rate. An initial decrease followed by a subsequent significant increase compared to the non-BLM-treated controls was observed in all parameters studied except the mitotic rate, which remained lower than in the control animals during all 48 hours. The transit time of the cells through the S-phase was initially slightly prolonged, but thereafter it seemed to be shorter than that of the controls. BLM seems to provoke a partial blocking of cells in the G1 phase. When the block is released, a greater number of cells pass through the S phase in partial synchrony at a higher than normal speed. BLM induced a low mitotic rate which remained below the level of that of the normal animals after stripping, even though there obviously was a considerably higher influx of cells from the S phase to the G2 phase. This resulted in a subsequent accumulation of cells in the G2-phase. Thus, BLM has also a blocking effect on the G2-M boundary of the cell cycle. This inhibitory effect of BLM on the mitotic rate was shown to be independent of the effect of BLM on the DNA synthesis. BLM therefore seems to have complex influence on epidermal cell kinetics in vivo. Cells in G1-phase are partially and transiently blocked, but this block is soon released. These cells thereafter pass through the S-phase and pile up in the G2-phase, because BLM also blocks the passage of cells from the G2-phase to mitosis. The overall reduction in cell proliferation seen after BLM in vivo seems mainly to be due to the effect on the G2-M boundray of the cell cycle.

[Age profile of the antihemagglutinins and antineuraminidase antibodies in type A influenza (author's transl)].

Antineuraminidase and antihemagglutinating antibody studies were carried out in parallel in sera from subjects born in Bulgaria in 1968-1972, 1956-1960, 1946-1950, 1925-1935 and 1917-1920. It was found that the amount of both antineuraminidase and antihemagglutinating antibody in sera from normal subjects could vary depending upon the year of birth and the strain used for the test. The antibody spectrum was most narrow in children under 4 and wider in subjects born before 1925. Serograms of the age distribution of antineuraminidase antibody were different in 4 out of 5 strains used which confirmed the hypothesis of Francis and Davenport of the more solid immunological response to the first exposure to virus. Parallel examinations of the antibody spectrum in human sera confirmed differences in the antigenic specificity of the "active center" of neuraminidases of A/Hong Kong/1/68 (H3N2) and A/Singapore/1/57 (H2N2) viruses and for the first time provided evidence attesting to the existence of similar strain differences in neuraminidases belonging to the N1 group. Examinations for antineuraminidase antibody in addition to antihemagglutinins increase the effectiveness of evaluation of the immunological structure of the population.

Clinical and haemostatic parameters related to thromboembolism and low-dose heparin prophylaxis in major surgery.

In a study of 112 patients undergoing elective major surgery clinical and haemostatic data was followed in connection with a double-blind investigation on the effect of subcutaneous low-dose heparin prophylaxis. None of the patients developed severe thromboembolism but according to lung photoscanning and leg scanning 41 of the patients had deep vein thrombosis and/or pulmonary embolism. Clinically thromboembolism appeared within 4 days after operation. In 22 patients with epidural anaesthesia the incidence of thromboembolism was lower than in the patients with general anaesthesia. The extension of the operation was positively correlated to a higher incidence of thromboembolism. The surgical trauma was reflected in most of the routine haemostatic laboratory parameters, hiding possible minor changes caused by subclinical thromboembolic complications. The low doses of heparin could only be detected with more sensitive methods. A comparison of sodium and calcium heparin administered subcutaneously revealed no significant differences.

Ultrastructural changes in scleromyxedema.

Skin biopsy specimens from a 60-year-old patient with paraproteinemia and generalized changes of the skin typical of scleromyxedema were studied with the electron microscope. The dermis was dominated by collagen fibrils and accumulations of peculiar connective tissue cells, while elastic tissue was sparse and in some areas completely absent. Large, sharply demarcated areas of a filamentous material were occasionally observed. The collagen fibrils were often surrounded by thin filaments with a periodic segmentation and by many glycosaminoglycan (mucopolysaccharide) filaments. The elastic fibres contained large amounts of elastic fibrils and small amounts of a homogenous matrix. The cytoplasm of the above-mentioned cells was dominated by lysosomes in different stages of development, often occupying almost all the cytoplasmic area. The collagen fibrils were found in close relation to these cells, frequently inside invaginations of the cells. Furthermore, collagen fibrils were observed free in the cytoplasmic area and inside the lysosomes, indicating lysosomal degradation of collagen.

Stereometric observations on the hypothalamic neurosecretory system of the spotted owlet, Athene brama Temminck, in total preparations.

The configuration of the HNS of the spotted owlet, Athene brama Temminck, was studied by employing the bulk-staining technique. This enabled a three-dimensional view of the HNS. The neurons of the SON are distributed into three divisions, namely, the preoptic, median and lateral. The PVN is U-shaped; the distal portion of the arms of the U extends laterally to form the lateral division of the PVN, which remains in continuity with the lateral division of the SON. The periventricular division of the PVN joins ventrally the median division of the SON. Unlike in the passerine birds, in the spotted owlet the PVN is more prominently developed than the SON. Since the axonal pathways are not stained, the proximal portion of the hypothalamo-hypophysial tract could not be demonstrated in bulk-stained preparations. Hence it was not possible to trace with certainty the origin of the axonal terminations in the ME and in the NL. Anterior of the ME, the tract branches into two; one branch enters the zona externa to the ME and the other is continued into the NL. The ME is divisible into an AF-positive anterior region and an AF-negative posterior region. The NL is saccular and heavily loaded with the NSM. Studies on the HNS of the spotted owlet by the bulk-staining technique reveal that the general configuration of the system is comparable to that of the HNS of birds studied previously.

A quantitative study of the pregnancy zone protein in sera of woman taking oral contraceptives.

By means of a single radial immunodiffusion the concentration of "the pregnancy zone protein" (PZ) was measured in sera from women taking oral contraceptive drugs. Woman taking only 0.3 mg. of norethisterone were found not to induce measurable amounts of PZ, whereas women taking combined contraceptive drugs showed a significant rise in concentration of PZ. After six months' treatment, 1 mg. of norethisterone and 0.1 mg. of meastranol daily were found to give an average concentration of PZ amounting to 59 mg. per 100 ml., that is, approximately half of the concentration of PZ in sera from women during the last trimester of pregnancy. Different combined contraceptive pills gave rise to different concentrations of PZ.

Differentiation of the neural plate and neural tube in the young chick embryo. A study by scanning and transmission electron microscopy.

The differentiation of the presumptive neural plate, the neural plate and the neural tube have been investigated in the chick embryo by SEM, TEM and histochemical techniques. The relationship of these tissues to neighbouring structures, including extracellular materials, has also been studied. When SEM micrographs of primitive streak stage embryos were examined in stereo, it was found that cells which had been invaginating at the time of fixation were similar in shape to fibroblasts migrating in vitro. It was concluded that SEM stereo pairs could provide evidence about the mode and direction of cell migration. Many more mid-bodies have been found associated with the developing neural tissue than with the lateral ectoderm. It was found possible to recognise mid-bodies not only by TEM but also by SEM. It is therefore proposed that SEM montages may be used for assessing which regions of a tissue have recently undergone extensive mitosis. The beads on the specialised threads seen in the early stages of development are now considered to be formed from mid-bodies. Similar, but unbeaded threads have been described which span the gap between the neural folds just prior to the dorsal closure of the neural tube and it seems probably that these threads help to close the neural tube. It is suggested that the beaded threads arise by incomplete separation of two daughter cells at mitosis, whereas the unbeaded threads form by outgrowth of cell processes.

Endodontic education.

Undergraduate endodontology is taught in Australian Universitiies in Restorative or Conservative departments. Courses generally consist of twelve lectures, three seminars and technique courses. Average clinical experience is six to eight endodontic treatments in anterior and posterior teeth. Comparison with American and Scandinavian teaching, staff and facilities indicates that this subject is receiving considerably less emphasis in Australia. As endodontic therapy is eseential in any dental health programme based on preventive dentistry there is a considerable need to upgrade undergraduate endodontic teaching in Australia.

Studies of "potentially lethal damage" in EMT6 mouse tumour cells treated with bleomycin either in vitro or in vivo.

Studies have been carried out into the effect usually referred to as "repair to potentially lethal damage" following the treatment of cells with bleomycin. In vitro, increased survival was seen with delayed subculture of cells in both exponential phase and plateau phase. It was unimportant whether the medium present during the delay period had been previously used to support cell growth. Exposure of cells growing as a solid tumour in vivo to bleomycin (4 mg/kg), gave a surviving fraction of 2 X 10(-3) if assay was carried out at 30 min but a surviving fraction of virtually 100% if assay was delayed until 6 h. Various possible artefacts have been eliminated as reasons for the observations but doubts are raised regarding the nature of the mechanism involved.

[Electronmicroscopic and immunohistochemical studies on human lymphocytes].

Lymphocytes from the blood of healthy individuals and of patients suffering from CLL were investigated by electron microscopy and peroxidase-immunohistochemistry. B-lymphocytes were labelled by heterologous, peroxidase-conjugated antisera directed against the Id-determinants of their membranes. T-lymphocytes were labelled by an indirect method: specific incubation with a specific anti-T-cell-globulin from the rabbit; labelling-incubation with a peroxidase-conjugated anti-rabbit-IgG-globulin from the sheep. In addition, T-lymphocytes were identified by their ability to form rosettes with sheep erythrocytes spontaneously. The quantitative results were: about 80% T-lymphocytes and about 24% B-lymphocytes in normal persons, the opposite results in CLL. T- and B-lymphocytes were photographed electron microscopically; the number of organelles in the single cells was evaluated: lysosomes in the average are more numerous in T-lymphocytes, ergastoplasm in B-lymphocytes, mitochondria are equally distributed in both groups of cells. There is so much overlapping, however, that the single cell only with the aid of immunochemistry or rosette formation can be identified as a B- or T-cell. In both, the T- and the B-cell-series, different forms of lymphocytes can be distinguished according to the degree of cell differentiation. Some further problems, as specificity of the antisera and labelling of the cells by means of their Fc-receptor are discussed.

111Indium-bleomycin breast and axilla imaging.

111Indium-Bleomycin (111In-Blm), a new radiopharmaceutical, was administered intravenously to 37 patients with benign and malignant breast lesions. Early and delayed images of both the breasts and axillae were made, and results were correlated with physical examination, histopathology of the excised lesion, mammography, and thermography. In 18 patients with malignant disease, clinical examination of the breast and axilla correlated with histopathology in 78 and 54% of the cases, respectively. Images of the breast were accurate (true positives) in 83% of the cases. Images of the axilla were accurate in 62% of the cases. Mammography was correct and suggested malignancy in 88%, and thermography in 73% of the cases. In 19 patients with benign breast lesions, clinical examination of the breast and axilla correlated with histopathology in 68 and 95% of the cases, respectively. Scans of the breast and axilla were correct (true negative) 79 and 95% of the time, respectively. Mammography was correct, and suggested benignancy, in 53% and thermography in 25% of the cases. Imaging of the breasts using 111In-Blm appears to be as accurate as physical examination and mammography for palpable benign and malignant breast tumors. It is less accurate than mammography for microscopic malignancies. Axillary imaging does not appear to be worthwhile because many axillary metastases are too small for detection with current nuclear medicine instrumentation.

Potentiation of IgE-mediated cutaneous reactivity and blood leucocyte histamine release by deuterium oxide.

A previous study (Gillespie & Lichtenstein, 1972) demonstrated that there was potentiation of histamine release from human peripheral blood leucocytes following exposure to antigin or anti-IgE in deuterium oxide (D2O). The current study confirms the results with human leucocytes and indicates that the degree of histamine release due to anti-IgE or its potentiation by D2O appeared independent of the serum IgE concentration of the cell donor. Further studies demonstrated that the peripheral blood leucocytes from monkeys with a sufficient degree of IgE-mediated reactivity to Ascaris antigen-released histamine following exposure to that antigen. This leucocyte histamine release occurs in animals with immediate-type cutaneous and respiratory reactivity following challenge with this antigen. Peripheral blood leucocytes from certain monkeys release histamine following exposure to anti-human IgE. Both of these Rhesus leucocyte responses are potentiated by D2O. This potentiation of histamine release in vitro in two species by D2O was compared with the potentiation of cutaneous reactivity in IgE-mediated cutaneous reactions in two species. The addition of D2O to Ascaris antigen or anti-IgE increased the end-point cutaneous titres to these stimuli in Rhesus monkeys and the addition of D2O to Ascaris or ragweed antigen increased the end point cutaneous titre to these reactants in allergic dogs. D2O did not potentiate cutaneous reactivity to histamine in either dog or Rhesus monkey.

Maturation of the somatosensory evoked potentials in normal infants and children, with special reference to the early N1 component.

The early components of the somatosensory evoked potential (SEP) were analysed in 39 normal newborns in REM sleep, 35 normal awake children between 1 month and 9 years and 16 normal awake adults. Electrical pulses were delivered at random intervals to fingers of the contralateral hand. The SEP were averaged from several electrodes in the parietal scalp focus and several runs were compared to estimate precise latencies and durations (Fig. 5, 6, 7). The system bandpass extended to 3 kc and many precautions were taken to exclude interference. The limb temperature was carefully maintained at normal value in order to avoid undue slowing of peripheral conduction velocity. The SEP presented markedly increased latencies for near-threshold stimuli (Fig. 3) while becoming even more focalised (Fig. 1). Background data were obtained in order to standardize the parameters for the maturation study (Fig. 2, 3, 4). In the children, the early negative component was found to undergo progressive changes. It was only at the age of about 8 years that the adult pattern was approached (Fig. 8, 9, 10, 11). The duration of the early negative N1 component decreased quite progressively. The latencies to onset and to peak were also shown to follow a highly consistent pattern when the body length of the subjects was taken into account (Fig. 9, 10). By dividing the data by the body length, functions were obtained which could be said to apply to a "standard" individual whose body length did not change from birth to adulthood and remained at one meter. Such plots made it possible to appreciate the true extent of the SEP maturational changes from birth. The quantitative data thus provided should serve as a useful reference for subsequent studies on developmental changes of the brain and for clinical applications to diseased children.

Biological signal detection by the autocorrelogram and a recurrence frequency method.

The signal detection property of the autocorrelation function (ACF) is conventionally derived on the assumption of two conditions that may not be present in biological time sequences: (1) the measured variable is assumed to be the sum of a noise and a signal function; (2) the signal function is assumed to continue throughout the sequence. In neuroelectric sequences of all-or-none responses the first assumption would not be appropriate if most of the elements seen by the recording electrode were accessible to the signal input. The sensitivity of the ACF was investigated in simulated sequences in which the random noise was entirely replaced by a periodic square wave signal function for a small number of repetitions and the results compared with a recurrence frequency function (RFF) developed during this work; under these conditions the ACF proved to be significantly inferior. Sensitivity of the ACF was the same for a pure signal function as for an additive function of signal and noise but sensitivity of the RFF was severely degraded for the latter. A variety of neuroelectric sequences was studied and many were found in which strong rhythmic processes revealed by RFF were not detected by the ACF. It is suggested that both methods be used when the signal function may be present during only a small part of the data-collection period and when it is not known to what degree the measured variable may be the sum of a noise and signal function. When an additive function is known to exist, the ACF will be the method of choice; for pure signal functions REF will be more sensitive.

Study of contingent negative variation and the post-imperative wave in the presence of interference.

The aim of this investigation was to study ways of inducing prolonged contingent negative variations (CNV) in human subjects. Fourteen subjects underwent the following experimental procedure: 1st day: 2 CNVs in control conditions were recorded (16 successive trials, each with standard S1--S2 paradigm). 2nd day: 5 successive CNVs were recorded:(1) control; (2) labyrinthine stimulation before 1st, 5th, 9th and 13th trials with simple mental calculation performed between S1 and S2, i.e. during CNV; (3) mental calculation only; (4) labyrinthine stimulation only; (5) control. None of the three interference situations caused any significant changes in the mean and maximal CNV amplitudes measured between S1 and S2. On the other hand, the post-imperative part of the CNV was significantly (P less than 0.025) prolonged but only in the situation where 2 types of interference were applied at the same time. This increase disappeared with removal of the double interference. This change, which matches that observed spontaneously and reversibly in certain mental patients in the acute phase of their illness, seems to reflect a transient alteration of the psychophysiological state of the subject. The possibility of inducing these changes in a normal subject could serve as a means of determining the individual threshold of stress.

[Studies on juvenile chronic hepatitis].

Follow-up study of 40 children suffering from chronic hepatitis. The diagnosis was made by liver needle biopsy with the Menghini method, when clinical signs or laboratory data of liver disease had lasted for more than 6 months. 24 patients showed the histological pattern of the aggressiv type of chronic hepatitis according to the definition of the European Association for the Study of the Liver (1968). In this group only 5 children had autoantibodies in the serum (so-called lupoid hepatitis). The HBAg positive courses played the most important part in the chronic persistent group as well as in the aggressive one. According to literature only the patients with the aggressive type have been treated with prednison, because chronic persistent hepatitis has a good prognosis without any treatment. In nearly all cases high transaminases and gammaglobulin levels decreased during the treatment with prednison, whereas the histological signs of inflammation seldom changed. Cirrhosis of the liver has developed in 2 HBAg positive patients of the aggressive group, who had not consequently received their daily dose of prednison.

Tumor-bound immunoglobulins. Evidence for the in vivo coating of tumor cells by potentially cytotoxic anti-tumour antibodies.

The experiments described herein were designed to determine whether part of the Ig coat of tumor cells consists of specific anti-tumor antibodies. It was demonstrated that the inoculation of polyoma virus-induced sarcoma cells (SEYF-a) into syngeneic A.BY mice stimulates the production of cytotoxic antibodies against the tumor-cell population. The level of these antibodies, which was undetectable during the first week after transplantation, increased markedly during the second week, and remained high thereafter. Following the increase in cytotoxic antibodies in the serum, a cell-bound potentially cytotoxic antibody was detected on the tumor cells by testing their sensitivity to rabbit complement. The increase in cell-bound, potentially cytotoxic antibody followed the kinetics of the increase in serum antibody during the second week after transplantation and was inversely correlated to the amount of free antigens on the cell surface. These antigens, responsible for the sensitivity of the cells to a syngeneic hyperimmune cytotoxic antiserum, became non-available for the cytotoxic antibodies during propagation of the tumor cells. Cells from a tumor propagated for 3 weeks could not compete for anti-tumor antibodies with cells propagated for 1 week. Yet it was possible to increase the antigenic capacity of cells from an old tumor by a treatment that would cause the release of tumor-associated Ig. Cytotoxic anti-SEYF-a antibodies could be dissociated from tumor cells propagated in vivo by methods causing dissociation of antigen-antibody complexes, and detected in tumor eluates.

Sudden interruption of leaflet opening by ventricular contractions: a mechanism of mitral regurgitation.

The motion of both mitral cusps and the presence of valvular regurgitation during ventricular contractions were investigated in seven experiments on dogs in which radiopaque markers had been sutured to the cusps and the valve annulus 1-32 wk before the studies. Cineangiograms of the left ventricle were obtained during ventricular ectopic beats, interposed throughout the cardiac cycle (20-99% of cycle length) and during induced variations in the P-R interval (0-200 ms). Mitral regurgitation was observed only during a) weak, early ectopic beats (peak pressure below 34 mmHg) which were incapable of closing the cusps and b) when ventricular contractions suddenly interrupted normal leaflet motion toward the ventricle, during three well-defined periods of diastole (diastolic valve opening, diastolic rebound, and atrial opening). Valve closure following sudden reversal of cusp opening was slow and the leaflets often did not arrive simultaneously at their closed positions. These findings suggest that sudden interruption of leaflet opening by ventricular contractions is an important mechanism of transient mitral regurgitation in the normal heart.

Reagin-mediated asthma in rhesus monkeys and relation to bronchial cell histamine release and airway reactivity to carbocholine.

Rhesus monkeys with persistent immediate-type cutaneous and respiratory responses (RR) to ascaris antigen (AA) were compared with rhesus monkeys with skin reactivity and no respiratory responses, and animals with no skin reactivity and no respiratory responses to inhaled antigen (NR). The RR group could not be distinguished from the nonresponding (NR) group by the cutaneous skin test titers, serum, or respiratory secretion IgE concentration. Leukocyte histamine (H) release due to anti-IgE was similar with peripheral blood leukocytes and bronchial lumen mast cells (MC) from RR and NR animals. The RR group of animals could be distinguished from the NR group by their degree of sensitivity to inhaled carbocholine and H release from respiratory MC exposed to AA. The RR group demonstrates consistent, persistent respiratory responses suitable for immunologic, pharmacologic, and physiologic studies. Finally, it was found that the IgE concentration in respiratory secretions of rhesus monkeys was comparatively higher than in serum, evidence for IgE as a secretory Ig in the respiratory tract of this species.

In vitro studies of the genetically determined unresponsiveness to thymus-independent antigens in CBA/N mice.

The X-chromosome-linked B lymphocyte defect of CBA/N mice has been studied in vitro by comparing the ability of (CBA/N X DBA/2)F1 (X-/X- X X+/Y) male (X-/Y) and female (X-/X+) spleen cells to respond to the thymus-independent antigen DNP (or TNP)-AECM-Ficoll. (CBA/N X DBA/2)F1 male spleen cells failed to generate significant in vitro anti-TNP antibody responses to DNP- or TNP-AECM-Ficoll, in contrast to spleen cells from F1 female (X-/X+) mice which responded normally to these T-independent antigens. Spleen cells from male F1 mice responded almost as well as F1 female cells to the thymus-dependent antigen, TNP-sheep red blood cells (TNP-SRBC) in vitro. Adding F1 male cells to F1 female cells failed to reduce the response of the latter to DNP-AECM-Ficoll, suggesting that the inability of F1 male cells to respond was not due to active suppression. The response of F1 male spleen cells to TNP-SRBC was not impaired by adding high concentrations of TNP-AECM-Ficoll indicating that the mechanism of unresponsiveness was not tolerance induction in all TNP-specific precursors. Lymphocytes from F1 male mice were capable of forming anti-TNP antibody after stimulation with lipopolysaccharide (LPS) in high concentrations; DNP-AECM-Ficoll had no effect on this polyclonal response. B lymphocytes from mice bearing only the X-chromosome of the CBA/N strain thus display a profound defect in B cell activation. This functional defect may represent either an inability of the defective B cells to be activated by thymus-independent antigens or the absence of a sub-class of B cells which respond to thymus-independent antigens.

Properties of antigen-specific suppressive T cell factor in the regulation of antibody response of the mouse. II. In vitro activity and evidence for the I region gene product.

An antigen-specific suppressive T cell factor, which was extracted from carrier-primed T cells, was further characterized in an in vitro secondary antibody response. The factor was capable of suppressing secondary IgG antibody response of primed spleen cells when it was added to the culture together with relevant antigen. The suppressive T cell factor was not released from primed T cells by a short-term culture with antigen, but was kept bound to the membrane of the residual cultured cells, only the physical disruption of which can release the T cell factor. The target of the suppressive T cell factor was determined as being the helper T cell, since the factor did not exert any effect in the absence of the helper T cell with identical specificity to that of the factor. The suppressive activity was completely absorbed with alloantisera specific for products of the I region of H-2 complex, although various anti-immunoglobulin antisera failed to do so. Close analysis of the specificity of alloantisera capable of absorbing the suppressor molecule indicated that the suppressive T cell factor may, in fact, be an I region gene product probably coded for by genes in I-A and/or I-B (including I-E) subregions.

Detection of alloantigens during preimplantation development and early trophoblast differentiation in the mouse by immunoperoxidase labeling.

An immunoperoxidase-labeling technique allowing visualization of antibody binding to the cell surface at the electron microscopical level has been employed an an analysis of H-2 and non-H-2 alloantigen expression on the early mouse embryo. The presence of non-H-2 antigenic determinants has been confirmed on eight-cell, morula, and blastocyst stages of development. Contrary to previous reports, however, low levels of H-2 antigen have also been detected on the blastocyst. This is the earliest stage at which H-2 has been shown to be expressed on the fertilized mouse egg and may reflect the greater resolution of the immunoperoxidase technique. Using two different models to study the critical peri-implantation stages, those of experimentally induced blastocyst activation and blastocyst outgrowth in vitro, it has been demonstrated that antigen loss occurs on the trophectoderm at the time of implantation, and that this is not necessarily dependent upon maternal influence. It is suggested that the loss may be an important factor in the prevention of maternal immune rejection during the establishment of the fetal allograft. The two major components of the early postimplantation conceptus display a striking differential in antigenic status. The embryonic sac shows a high degree of peroxidase labeling, while the ectoplacental cone trophoblast is unlabeled. These findings add support to the concept of antigenic neutrality of the early trophoblast and its role in the maintenance of a normal fetomaternal immunological equilibrium.

Structural characteristics of the alloantigens determined by the major histocompatibility complex of the guinea pig.

The GPLA B and Ia (I region-associated) antigens are the products of genes found in the guinea pig major histocompatibility complex. Because of their importance in immune response phenomena, a structural study of these antigens was undertaken. [3H]leucine and [3H]fucose were internally incorporated into guinea pig lymph node cells. The GPLA B and Ia antigens were solubilized by the nonionic detergent Nonidet P-40, purified by affinity chromatography using an adsorbent column of lentil lectin, isolated by immunoprecipitation, and examined by discontinuous polyacrylamide-sodium dodecyl sulfate gel electrophoresis. The GPLA antigens B.1, B.2, B.3, and B.4, were shown to be glycoproteins of mol wt 40,000 daltons and to be noncovalently associated with a 12,000 dalton protein. The molecules bearing B.2 and B.3 in a B.2/B.3 heterozygote are shown to be separable, suggesting the antigenic determinant is a primary gene product. In addition, a new GPLA determinant, S, which resembles the B antigen in that it is found on a molecule of approximately 40,000 daltons, was studied. In a B.2/B.3 S+ animal the molecule bearing antigen S was shown to be independent of those bearing B.2 and B.3, providing evidence that the genes determining B and S are at separate loci. The Ia-bearing molecules identified by anti-Ia.2,4 are glycoproteins of mol wt 58,000 daltons which are composed of two subunits of 33,000 and 25,000 daltons, respectively, linked by disulfide bonds. The Ia-bearing molecules are independent of GPLA-bearing molecules, indicating different loci determining these antigens. By all criteria, the guinea pig GPLA B antigens appear homologous to the murine H-2D and H-2K antigens, while the guinea pig Ia antigens appear homologous to the Ia antigens of the mouse.

Clinical observations of juvenile nonprogressive muscular atrophy localized in hand and forearm.

Twenty-seven patients with juvenile nonprogressive muscular atrophy localized in the hand and forearm were analyzed. The clinical characteristics were juvenile male occurrence, insidious onset, specific distribution of localized muscular atrophy and a stationary course. On electromyography, denervation voltage (or giant NMU) is found in the atrophied muscles and sometimes in contralateral nonatrophied ones. Sensory disturbance was not remarkable. Although the etiological factor was not known, strenuous exercise of arms in sports was noted frequently in the history.

Evaluation of results of thymectomy in myasthenia gravis.

The results of thymectomy carried out in 150 cases of myasthenia gravis are discussed. In a group of 123 cases followed for 1 to 5 years after the operation, full remission was observed in 24.4% of cases, significant improvement in 36.6%, slight improvement in 24.4% and no improvement in 8.1%, while deterioration occurred in 1.6% of patients. No correlation was found between the result of the operation and the age and sex of patients, but better results were achieved in those treated surgically rather soon after the onset of symptoms. This correlation was particularly evident in the group with full remissions. The results obtained in the cases without thymic tumors were better than in the cases with tumors. No correlation was noted between the results of the operation and the histological characteristics of the thymus in the group with thymic hyperplasia and in the group with thymic atrophy. The surgically treated group (150 cases), compared with the conservatively treated group (75 cases), showed the superiority of the surgical method (lower rate of death and deterioration, higher rate of improvement and remission). In discussing the indications for surgical treatment the authors emphasize that advances in anaesthesiology in recent years have reduced the risk of operation. It is suggested that the indications for surgical treatment should be expanded and operations should be performed as early as possible after the onset of clinical manifestations without regard to the age and sex of the patient. Operation should not be considered in cases belonging to group 1, 2a (sometimes 2b) only, with duration of the disease over 8-10 years and with little or no progression of the process, if the presence of a thymic tumor has been excluded.

3H-thymidine autoradiography of the CSF cells in cases of non-neoplastic disease.

Human CSF cells in cases of non-neoplastic disease of the central nervous system (CNS) were examined in vitro by 3H-thymidine autoradiography. Immediately after withdrawl by lumbar or ventricular puncture, the CSF was incubated in a sedimentation chamber at 37 degrees C for 1 hr with an admixture of 3H-thymidine at a concentration of 1-2 muCi/ml CSF. In a few cases the CSF withdrawn was incubated in a glass tube in the same condition as in a sedimentation chamber, and the CSF cells were collected by centrifugation. The CSF cells collected were fixed in methanol and the microautoradiographic procedure was performed. Labeled CSF cells were found in 21 cases out of 22. The average labeling index of the total nucleated cells was 0.22% with the highest labeling of 0.74%. Almost all the labeled cells were thought to be medium to large sized lymphocytes and monocytoids. Peripheral blood was examined by a similar method and the results were compared with those of the CSF. It may be noteworthy that thre exist DNA synthesizing cells in the CSF even in a non-neoplastic state of the CNS, although the number is not large.

[The EEG after frontal and frontobasal skull and brain injury (author's transl)].

Eighty-eight patients with confirmed frontal and frontobasal lesions were examined. The EEG changes were investigated according to type and frequency both after recent trauma and in the later stages (Fig. 1). The general changes showed mainly slight (Fig. 2B) and only rarely severe, dysrhythmias (Fig. 3A) which, however, were frequently associated with paroxysmal outbursts. Focal findings were largely temporal and frontal in the second place (Fig. 2A). All pathological EEG changes showed a tendency to improvement, so that in the late phase they occurred rarely and less severe (Fig. 4). They showed no essential difference from the EEG of uncomplicated craniocerebral traumata. The relations between the EEG and the clinical form of craniocerebral trauma were examined. In contusions of the head the normal EEG was predominant. The pathological EEG was prominent with cerebral concussion and even more so with cerebral contusion. On the other hand, there was no relationship between the EEG and the type of fracture. A significant predominance of pathological EEG was found only in frontobasal fractures with rhinorrhoea. The EEG changes, classified as focal and dysrhythmic, show no statistically significant relationship either to the form of craniocerebral trauma or to the type of fracture. The effect of force coming form in front has apparently been used up chiefly in producing the frontal or frontobasal fractures, and leads to relatively slight cerebral damage. The clinical significance of the EEG does not actually lie in the diagnosis of these injuries, but, as always, in the establishment of the severity and extent of the cerebral lesion and the detection of complications.

A randomized pilot study comparing two regimens in the treatment of squamous cell carcinoma.

Thirty-five patients with epithelial carcinomas not considered to be surgically resectable were randomized to receive two different chemotherapy regimens. Regimen 1 was CCNU followed by bleomycin, and Regimen 2 was a combination of CCNU, bleomycin, methotrexate, and vinblastine. Five of 14 patients treated with CCNU-bleomycin had partial responses. Three of 15 patients treated with the four-drug combination had a partial response. The toxicity of the four-drug regimen was significantly greater than that of the two-drug regimen, while the response rate was greater among those patients treated with two drugs. No significant clinical infections occurred despite the fact that leukopenia and thrombocytopenia were more severe and frequent with the four-drug regimen. Most importantly, the two-drug regimen is well tolerated as an outpatient procedure. Two of the two-drug and one of the four-drug recipients were converted from an inoperable to an operable state. In view of the fact that there has not heretofore been an effective chemotherapeutic regimen for Stage III and IV squamous carcinoma, this is a significant observation. It is of note that patients with squamous cell carcinoma of the head and neck were those who responded best to this treatment, although the treatment is worthy of consideration for advanced squamous carcinoma in other areas.

Treatment of attacks in hyperkalaemic familial periodic paralysis by inhalation of salbutamol.

In fifteen patients with hyperkalaemic familial periodic paralysis, inhalation of salbutamol alleviated hyperkaleamia and paralysis precipitated by exercise or oral administration of potassium chloride. In-vitro studies with rat soleus muscles indicated that the hypokalaemic effect of salbutamol is related to stimulation of the active coupled transport of sodium and potassium in muscle cells. Follow-up studies proved that the inhalation of salbutamol is a simple and adequate method for the treatment of the paralytic episodes in these patients.

Cerebral thrombosis, cerebral haemorrhage, and ABO blood-groups.

The ABO blood-group distributions of 1460 patients who had died from a stroke were compared with those of a control group of 20 705 controls selected at random from the healthy population at risk (i.e., over thirty-five years of age and matched for age and sex ratio). The cause of death was certified as cerebral thrombosis in 329 cases and as cerebral haemorrhage in 482 cases, these diagnoses being established in neurological hospitals; the remaining 649 cases had an unspecified type of stroke, the diagnosis being made by general practitioners. In the group with unspecified type of stroke the blood-group distribution was practically the same as the distribution in the controls. In the thrombosis cases there was an excess of blood-groups A and AB and a deficiency of O and B; in cerebral haemorrhage this situation was reversed. However, these were only trends; the differences were not significant at the 5% level. A statistically significant difference did emerge when the A+AB excess in thrombosis was contrasted with the O+B excess in haemorrhage, suggesting that this difference might be accounted for the major A subgroup (A1) and, consequently, A1B.

Menstrual blood-loss with intrauterine devices.

The effect of three intrauterine contraceptive devices (I.U.D.)-Lippes D, Dalkon Shield, and Copper 7-on menstrual blood-loss has been studied serially by objective methods in 279 women. All the women had a minimum of two cycles following delivery, abortion, cessation of lactation, or previous pill or I.U.D. use. All pads and tampons used for two further menstrual cycles before and for 12 cycles after I.U.D. insertion were collected, and the blood-loss was measured by extracting the haemoglobin by conversion to alkaline haematin. Mean menstrual blood-loss increased with all three devices. The amount of loss, the percentage of women losing more than 80 ml, the decline in haemoglobin concentration, and the incidence of anaemia during the 12 cycles following insertion were all greater among users of the Lippes Loop than of the Copper 7 with generally intermediate values for Dalkon Shield users. The mean increases in blood-loss were: for parous women fitted with the Lippes Loop, 48 ml, with the Dalkon Shield, 34 ml, and with the Copper 7, 18 ml; for nulliparae fitted with the small size of Dalkon Shield, 27 ml, and, with the Copper 7, 19 ml.

Is multiple sclerosis an age-dependent host response to measles?

Several lines of evidence support the possibility that multiple sclerosis (M.S.) may be an age-dependent host response to measles. In animals, measles evokes different responses depending upon age at inoculation. In man, measles is already known to produce at least two age-dependent responses: risk of subacute sclerosing panencephalitis is increased among those who have had measles before 2 years of age and risk of measles encephalitis increases with age, at least during adolescence. Studies of immigrant populations indicate that an event at or before adolescence but not infancy affects risk of M.S. In the tropics where M.S. is rare, measles tends to be acquired very early in life, usually before the age of 3, whereas in temperate areas, measles tends to be acquired later, after the age of 5. A retrospective study has shown that M.S. patients tend to have had measles later than controls. Mechanisms which might underlie an age-dependent host response to measles include maturation of an immune system (k.g., number of available B cells) or change in the metabolic state of a target cell (e.g., oligocytes which change from laying down myelin to maintaining it). If the hypothesis that M.S. is a host response to later measles infection is valid, then mass measles vaccination programmes should produce a decline in the rate of M.S., but the effect may not be discernible before 1980.

HLA matching and corneal grafting.

A series of 200 cases of full-thickness corneal allografts have been followed to determine whether HLA and ABO incompatibility influence prognosis of the grafts. 85% of patients with avascular corneas had clear, functioning grafts one year after transplantation. Only 33% of patients with severely vascularised corneas had successful grafts one year after transplantation. A significant association was found between severe vascularisation of the patient's cornea and irreversible graft rejection. In this group of patients, the proportion of grafts functioning was found to be ranked according to the number of HLA antigens shared by graft donor and recipient. Patients receiving grafts matching for 2 HLA antigens showed a failure-rate due to irreversible rejection of 26% at one year, in comparison with 57% and 62% of grafts matching for 1 or 0 HLA antigens respectively. ABO incompatibility or ABO phenotype of the recipient did not influence graft prognosis. The results indicate that patients with severely vascularised corneas should receive HLA-matched corneal grafts. The institution of HLA-typed cornea "banks" for treatment of such patients is advocated.

Psychiatric screening in general practice. A controlled trial.

This study reports the efficacy of the General Health Questionnaire (G.H.Q.) in the secondary prevention of minor psychiatric illness in a primary-care setting. 1093 consecutive attenders at a general practitioner's surgery were screened for minor psychiatric disorder using the G.H.Q. 32% were found to have a conspicuous psychiatric disorder and a further 11% had a hidden psychiatric disorder. The group with hidden disorders were randomly assigned to a treated index group and an untreated control group. The effects of case detection and treatment were beneficial and immediate, with the duration of episode of the disorder being much shorter for patients whose disorder was recognised by the general practitioner. For patients with more severe disorders there are significant differences still demonstrable between the groups one year later; but patients with mild disorders do equally well, some recovering spontaneously but others becoming manifest to the general practitioner over the next year and so receiving treatment. The "detected" group of patients increased their consultations for emotional complaints over the next year, but their total consultation-rate was not increased.

Vibrio cholerae flagellar antigens: a serodiagnostic test, functional implications of H-reactivity and taxonomic importance of cross-reactions within the Vibrio genus.

Serodiagnostic tests for all serotypes of Vibrio cholerae using H-antisera were investigated. Activity motile cell lines of 155 stock and international reference cultures of human, animal, fish, and halophilic Vibrios, Aeromonas, Comomonas, Pseudomonas, Salmonella, and Escherichia were investigated. Without exception, all cholera vibrios (including the NAG serotypes) reacted with H sera. Positive reactions were obtained specifically (a) within 2 hrs at 52 degrees C in the tube test using thick formalized suspensions and H antisera at optimal proportion titre and (b) within 30 sec by slide agglutination of fresh cultures. The other vibrios investigated reacted similarly with their homologous H antisera. 2. The rapid diagnostic techniques of fluorescent antibody labeling or immobilization were unsuccessful, V. cholerae flagella being refractive to H sera in these tests. V. cholerae was, however, sensitive in a type-specific manner to O antisera. These and related observations suggest that O antigen has a functional role in Vibrio motility. 3. Interspecies H cross-reactions between V. cholerae and fish and animal vibrios which correlated with bacteriologic similarity, were demonstrated. O antigens of these vibrios were strain specific. Cross-absorption analysis indicated that the H antigens of vibrios were characteristic and homogenous within the species, and therefore a potentially important taxonomic criterion of Vibrio species.

Banding technique used for the detection of chromosomal aberrations induced by radiation and alkylating agents TEPA and epichlorohydrin.

Blood samples from two healthy donors were exposed, (1) to 200 R of X-rays in G0 and G1S phases of the cell cycle, and (2) to epichlorohydrin 10(-6) M and TEPA 10(-4) M in G0 and/or in G1S and G2 phases. Part of the cells was processed for chromosome studies conventionally and the other part by the trypsinization banding technique. Detailed chromosomal analysis showed that, after irradiation, 38.2% of aberrations in G0 and 18.7% in G1S phases escaped cytogenetic detection when the conventional technique was used. After exposures to TEPA and ECHH, 10.9% of aberrations were undectable in G0 and 3.3% in G1S and G2 phases. The distribution of chromosome breaks was non-random both after irradiation and after exposure to alkylating agents. However, it differed according to the mutagen used. Some chromosomal segments were broken significantly more frequently than the others (e.g. 9q12), some were resistant to breakage (e.g. the whole Y chromsome). The segments represented by G-negative bands were more fragile than the G-positive and G-variable segments.

[Immunologic parameters in acute lymphatic leukemias in the course of different types of chemotherapy (author's transl)].

In 2 groups of patients under treatment with acute lymphatic leukemia and undifferentiated leukemia, apart from the lymphocyte count, a number of immunologic parameters were examined during the various stages of treatment. These parameters were related to the type of chemotherapy as well as the stages of treatment. This paper deals with the values of lymphocyte counts, of gammaglobulin, IgG, IgM and IgA and the results of the DNCB skin-tests, as a parameter for cellular immunity. The determination of gammaglobulin using the acetate sheet electrophoreses, showed no significant change. However a significant reduction of the IgG was shown during the periods of prophylactic reinduction with Vincristin in the Pinkel VII scheme. IgM quantities were significantly raised in the initial stage before treatment and in all periodes of treatment both in the Pinkel VII group and in an earlier therapeutic group with methotrexate two weekly and Vincristin pulses every 6 weeks. IgA was raised in both groups before treatment but significantly reduced during maintenance and reinduction phases. About 50% of the P VII cases had negative DNCB skintests during maintenance therapy, while all long term survivors in the earlier methotrexate treatment group had positive skintests. The 2 following papers deal with the results of PHA stimulation and the B and T cell determinations.

Induced abortion: 1975 factbook.

This report presents an overview of current international data on induced abortion, primarily from the demographic and public health points of view. Statistical tabulations make up the major part of the report, with the text providing background information. Opening sections review definitions, sources of data, and concepts of statistical analysis. Major topics for which data are presented include the legal status of abortion (Table 1); incidence of abortion (Tables 2-20); incidence of repeat abortions (Tables 21-23); period of gestation and abortion procedures (Tables 24-27); incidence of abortion with concurrent sterilization (Table 28); complications (Tables 29-31); and mortality (Table 32). The relationships between abortion and contraception (Tables 33-35) and the effects of changes in abortion policies on trends in the numbers of legal abortions, illegal abortions, total induced abortions, and births are evaluated in the final sections (Table 36). Two technical questions are discussed in the appendix (Tables 37-38).

HL-A8 and LD-8a in patients with myasthenia gravis.

The frequency of HL-A8 in myasthenia gravis is markedly increased in women (60-80%) but not in men. The MLC determinant, LD-8a, is frequently associated with HL-A8. Of the 37 female MS patients, 15 were LD-8a positive (41%), whereas of the males only one of seven was LD-8a positive. The frequency of HL-A8 was 68% in women and 29% in men with the disease. We therefore conclude that the gene which is responsible for the increased susceptibility to myasthenia gravis in women and which is present in the MHS region, is more closely linked to the SD-2 than to the LD-1 locus.

[The effect of drugs on "sino-atrial conduction time" and on sinus-node automaticity in man].

The effect of atropine, propafenone, and disopyramide on sinus node automaticity and "sino-atrial conduction" was tested in normal patients and patients with the sick sinus-syndrome. "Sino-atrial conduction time" was estimated indirectly by the extrastimulus technique. Atropine (n = 11) caused a significant increase in heart rate in all patients. The sinus node recovery time was shortened in 10 patients. "Sino-atrial conduction time" decreased on an average 35% (P less than 0.01). Three patients with a sick sinus-syndrome demonstrated a change of the pattern of the postextrasystolic pauses indicating great improvement in sino-atrial conduction. Propafenone (n = 10) led to a significant prolongation of the sinus node recovery time by 17% and of the "sino-atrial conduction time" by 27%. Disopyramide (n = 8) had no significant influence on heart rate and "sino-atrial conduction time". Sinus node recovery time was not changed in 6 patients. However, in two patients with a sick sinus-syndrome a dangerous prolongation of the sinus node recovery time after application of disopyramide occurred. The results indicate that atropine enhances sinus node function and sino-atrial conduction. On the other hand, propafenone and disopyramide exert either a depressant influence on sinus node automaticity or on sino-atrial conduction.

Cellular localization of kallikreins in rat submandibular and sublingual salivary glands: immunofluorescence tracing related to histological characteristics.

Immunohistochemical localization of rat salivary gland kallikrein was related to glandular structures in tissue processed and stained by various methods. In the submandibular gland, most of the kallikrein was located to cytoplasmic granules of the granular tubules. Cells of the striated ducts showed a faint cytoplasmic staining with a bright luminal rim that occasionally was seen also in the excretory ducts. Minor amounts of kallikrein was found in the interstitial tissue. In the sublingual gland, kallikrein was found in the cytoplasm of the striated duct cells and as a luminal rim in the main ducts. Acini were negative in both glands. Fixation in Helly's fluid preserved cytoplasmic granules and was thus superior for intracellular localization of kallikrein, whereas ethanol fixation, due to absence of non-specific background staining, afforded the most sensitive method for detection of small amounts of antigen. In the submandibular gland, best identification of granular tubules and striated ducts was achieved with DMAB-nitrite staining for tryptophan and counterstaining with Mayer's haemalum on sections of tissue fixed in Helly's fluid. In the sublingual gland, the duct system was best demonstrated by haematoxylin-eosin staining.

Thyrotoxicosis with painless thyroiditis.

Four women and one man with painless subacute thyroiditis presented with hypermetabolic signs and symptoms. Thyroxine (T4) and triiodothyronine (T3) resin uptakes (T3R) were increased but the 24 hour radioactive iodine (RAI) uptakes were less than 1 per cent. Surreptitious use of thyroid hormone was excluded. The thyroid was enlarged in one patient and nontender in all. Exophthalmos was absent. The protein-bound iodine level was 1.1 to 9.5 mug/dl greater than the T4 level. The sedimentation rate was normal or minimally increased, and antithyroglobulin and antimicrosomal antibodies were undetectable. In one hospitalized patient 84 per cent of the administered dose of 131I was recovered in the urine within 48 hours (normal 64 per cent) excluding extrathyroidal uptake. In all subjects the T4 and T3R levels fell to normal or slightly below normal within one to four months. An increase in the 2 and 24 hour RAI uptake to minimally increased or high normal values and return of the T4 and T3R levels to normal occurred in four of five patients within six months. In one of these, the administration of thyroid-stimulating hormone (TSH) resulted in an appropriate increase in 24 hour RAI uptake from 14.9 to 37.1 per cent. One woman remained clinically hypothyroid for six months with a low T4 concentration (3.2 mug/dl), an elevated TSH level (48 muU/ml) and evidence of a persistent organification defect -- two hour RAI uptake decreasing from 33 to 23 per cent after the administration of perchlorate and the 24 hour RAI uptake increasing from 32 to 76 per cent following the administration of TSH. At 21 months after the initial onset of her illness, she is euthyroid but increased RAI uptake persists. The clinical course in four of the five patients is similar to that in an additional eight patients treated during the same time period who presented with typical subacute thyroiditis. Thus, these patients have a form of painless subacute thyroiditis which presents as thyrotoxicosis but is differentiated from it by a low RAI uptake and in whom recovery of thyroidal iodine trapping is the first indicator of recovery. The hyperthyroidism is self-limiting and should be treated conservatively.

Immunochemical localization of in vivo bound immunoglobulins in pemphigus vulgaris epidermis. Employment of a peroxidase-antiperoxidase multistep technique for light and electron microscopy.

A multi-step immunocytochemical method utilizing horseradish peroxidase (HRP) as an immunological marker was employed to demonstrate, at the ultrastructural level, IgG antiepithelial autoantibodies bound in vivo to pemphigus epidermis, and circulating IgG antibodies, using monkey oesophagus as substrate. To demonstrate IgG the following antisera were employed in sequential steps: Goat antihuman IgG; rabbit antigoat IgG; goat-anti-HRP-serum. After incubating with the final antigen, HRP, the latter was visualized with a cytochemical method. Results paralleled those obtained by immunofluorescence, the antiepithelial antibody being demonstrated at the sites of the intercellular spaces of the epidermis. Electron microscopic cytochemistry showed IgG in the surface coat of epidermal cells and in the intercellular space, both in the desmosomal and the interdesmosomal areas. These results confirm the results of a previous study but the localization of the antibody is more exact than that obtained with HRP-conjugates. The present method is more versatile and specific than HRP methods that utilize HRP-conjugated antibodies.

The comparative specificity of 3 oestradiol-binding proteins. Rat alpha-foetoprotein, rat liver 17beta-hydroxy steroid dehydrogenase and anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) antiserum.

1. The specificity of 3 oestradiol-binding proteins was studied. Two of these proteins are naturally occurring (rat alpha-foetoprotein and rat liver microsomal 17beta-hydroxy steroid dehydrogenase) and the third is an artificially induced model, anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) gamma-globulins. 2. A specific binding procedure for each protein model permitted a determination of its affinity for oestradiol and for 30 other steroids. 3. The results obtained have brought to light the different areas of the steroid molecule that are important for its recognition by each of the three proteins. The two naturally occurring proteins (alpha-foetoprotein and 17beta-hydroxy steroid dehydrogenase) recognize the edge of the steroid defined by C-4, C-6, C-8 and C-15. On the other hand, the gamma-globulins recognize the opposite edge, i.e. that defined by C-2, C-10, C-11 and C-17. 4. Diethylstilboestrol, whose structure is analogous to that of a steroid, is only recognized by the two naturally occurring proteins.

Histamine release in human subjects by modified gelatin (Haemaccel) and dextran: an explanation for anaphylactoid reactions observed under clinical conditions?

Histamine release by modified gelatin (Haemaccel) and dextran (Macrodex) has been demonstrated in volunteers by direct and indirect methods. In a pilot study of Haemaccel, histamine release was observed in six of seven volunteers. The highest plasma histamine concentration was 4.8 ng/ml, the lowest 1.7 ng/ml: two of the subjects showed slight allergic reactions. Using Haemaccel batch 2551, 10 out of 12 subjects reacted to the rapid infusion of Haemaccel with increased plasma histamine concentrations, whereas none reacted to Ringer's solution. None of the 10 subjects had an allergic reaction, but an increase in gastric secretion was observed in eight. Changes in the venous basophil granulocyte count were found in both those who reacted and those who did not react to Haemaccel. After the rapid infusion of dextran the highest plasma histamine concentration was 5.0 ng/ml, the lowest 1.3 ng/ml. The withdrawal of blood had no influence on plasma histamine concentration. The incidences of histamine release produced by Haemaccel varied with different batches. Thus, it seems unlikely that immunological mechanisms are principally responsible. Nine instances of allergic and anaphylactoid reactions to plasma substitutes have been reported, seven after Haemaccel infusion, and two after dextran administration. One of the patients who received dextran died. Histamine release was always associated with Haemaccel infusion and corresponded in extent to the clinical symptoms observed, but there was no significant histamine release associated with the reactions to dextran.

Studies on the isolation and partial characterization of apolipoprotein D and lipoprotein D of human plasma.

This report describes further studies on the characterization of apolipoprotein D (ApoD), a recently recognized human plasma apolipoprotein, and presents results on the isolation and distribution of its lipoprotein form, lipoprotein D (LP-D). ApoD, isolated by a procedure combining hydroxylapatite and Sephadex G-100 column chromatography, migrated on 7% polyacrylamide gel as a single band with a mobility intermediate between those of A-II and C-II polypeptides. On double diffusion and immunoelectrophoresis, ApoD reacted only with antiserum to ApoD. It was characterized by the presence of all common amino acids including half-cystine. The amino terminal acid was blocked. Carbohydrate analysis demonstrated that ApoD is a glycoprotein with glucose, mannose, galactose, glucosamine, and sialic acid accounting for 18% of the dry weight of ApoD. The estimated molecular weight of ApoD IS 22 100. ApoD occurs in the serum as a lipoprotein which was isolated from high density lipoproteins3 by two different chromatographic procedures. In the first procedure, high density lipoproteins3 were treated with neuraminidase and chromatographed on concanavlin A. The retained fraction containing LP-D was purified by hydroxylapatite column chromatography. Alternatively, LP-D was isolated by a procedure combining chromatography of high density lipoproteins3 or whole serum on an immunosorber containing antibodies to ApoD, and hydroxylapatite column chromatography. LP-D displayed a single, symmetrical boundary in the analytical ultracentrifuge and a single band on 7% polyacrylamide gel electrophoresis. When injected into rabbits it produced antisera that reacted only with ApoD. On immunoelectrophoresis LP-D had a mobility different from that of lipoprotein A (LP-A). A direct immunological comparison of LP-D and LP-A showed a reaction of nonidentity. LP-D consists of 65-75% protein and 25-35% lipid. The lipid moiety contains cholesterol, cholesterol ester, triglyceride, and phospholipid. The phospholipid. composition is characterized by a relative high content of lysolecithin and sphingomyelin and a relatively low content of lecithin. We have concluded from these studies that ApoD is a unique apolipoprotein that exists in the form of a distinct lipoprotein family with a macromolecular distribution extending from very low density lipoproteins into very high density lipoproteins, but with a maximum concentration in high density lipoproteins3 and a minimum concentration in high density lipoproteins.

Gonadal and extragonadal yolk sac carcinomas: a clinicopathologic study of 14 cases.

Fourteen cases of yolk sac carcinoma, 10 occurring in gonadal, and four in extragonadal sites, seen at the Indiana University Hospitals from 1949 to 1974, were analyzed with respect to pathologic features, laboratory findings, and clinical course. Their histologic appearance was similar regardless of the site of origin. Two basic histologic types were observed--the more common endodermal sinus pattern and the rare polyvesicular vitelline form. The prognosis is unfavorable, but three of our cases exhibited objective responses to chemotherapy. In our small series, the better prognosis of testicular yolk sac carcinomas in children found by some authors was not evident. Four of the 6 patients with yolk sac carcinoma in which serum alpha-fetoprotein determinations were performed showed positive results. Three of these cases had residual or metastatic disease clinically. The demonstration of alpha-fetoprotein in the serum of patients with yolk sac carcinoma lends further support to the yolk sac origin of these tumors and could also prove to be of prognostic value by indicating the presence of residual or recurrent disease.

In vivo and in vitro measurements of the relationship of human squamous carcinomas to herpes simplex virus tumor-associated antigens.

An additional 244 unfiltered sera have now been studied in a series of controlled, coded tests to determine the relationship of squamous carcinomas of the head and neck and cervix to the presence of complement-fixing antibodies to herpesvirus-tumor-associated antigens (HSV-TAA) in both tumor-bearing and cured patients. Ninety % of sera from patients with squamous carcinomas had antibodies to HSV-TAA, in contrast to 11% of sera from patients with nonsquamous cancers and 4% of sera from noraml individuals. The temporal relationship of Stage 1 laryngeal carcinomas suggests that HSV-TAA appearance precedes the immune defects. An in vitro correlate of the previously demonstrated specific delayed hypersensitivity reactions in controlled skin tests of squamous carcinoma patients with HSV-TAA is reported. In leukocyte migration inhibition tests, the migration indices after incubation with HSV-TAA of peripheral blood leukocytes from patients with squamous carcinoma (x = 0.847) were in definite contrast to migration indices seen for normal leukocytes (x = 1.037) and patients with nonsquamous solid cancers (x = 1.03). Thus, these polypeptides elicit both humoral antibody response and cell-mediated reactivity.

Molecular interactions of the combined effects of bleomycin and x-rays on mammalian cell survival.

The interactions between bleomycin and X-ray damage and repair have been examined in rat and human tumor cells. Bleomycin itself indices extensive DNA single-strand breaks but does not appear to inhibit the repair of X-ray-induced DNA single-strand breaks. Quantitative analysis of these interactions is complicated by the retention of active bleomycin within cells that remains capable of further DNA degradation even under the conditions of alkaline sucrose gradient cell lysis. DNA double-strand breaks and/or disruptions of DNA-lipid complexes also occur following bleomycin exposure. X-ray-induced excision repair replication is only minimally influenced by even high concentrations of bleomycin. A small amount of excision repair is demonstrable in nonirradiated cells treated with high concentrations of bleomycin consistent with repair of bleomycin-induced nucleotide damage in cellular DNA by a "cut and patch" repair mechanism. Repair of bleomycin-induced DNA single-strand breaks also occurs. The data indicate that bleomycin and X-ray damage are quite similar both in their induction and repair, but that lesions occur and are repaired independently. The enzymatic mechanisms appear similar in the two cell types despite substantial differences in their sensitivity to bleomycin.

Evoked potential decrements in auditory cortex. I. Discrete-trial and continual stimulation.

In experiment 1 cats were exposed to sets of clicks (trials) with 1 min inter-trial-intervals to determine if the effects of repetitive stimulation on potentials evoked in the auditory cortex would be cumulative despite discrete-trial stimulation. Evoked potentials were averaged to give one average evoked potential (AEP) for each trial for each electrode; there were four cortical electrodes per subject. To test for dishabituation pawshocks were given between trials 60 and 61. Subjects were paralyzed to insure stimulus constancy. The latency and peak-to-peak amplitude of each component of each AEP was measured; significant amplitude decremented; and decrements were more frequent in components with latencies greater than 15 msec. A few amplitude increments and latency changes were also observed...

Blink reflex in hemiplegia.

An electrophysiological study of the blink reflex was undertaken in 20 normal subjects and in 28 patients complaining of central facial palsy caused by unilateral hemispheral damage. In normal subjects, the latency, amplitude and organization of R1 and R2 responses are well known. Habituation of R2 responses occurred between 1 and 2 c/sec stimulation rate. R1 responses habituated at a higher stimulation rate (5 c/sec). In patients with unilateral hemispheral lesion, our results showed that changes in the blink reflex responses were bilateral. On the hemiplegic side the responses showed a decreased amplitude, while they were facilitated on the "normal" side. However, there was no change in latency of the two components of the reflex, on both sides. On the other hand, habituation of the late component occurred on the hemiplegic side for low stimulation rates: (0.5--1 c/sec), while on the "normal" side there was less habituation (3--4 c/sec), as compared with normal subjects. These results agree with those of experimental studies on cortical modulatory influences on brain-stem nuclei. They suggest a tactile origin of the two components of the blink reflex.

Immunofluorescent polar tips of Rhizobium japonicum: possible site of attachment or lectin binding.

Rhizobium japonicum USDA 31 demonstrated marked polarity by binding homologous fluorescent antibody (FA) heavily on one end of the cell. FA prepared against R. japonicum strains 110 and 138, and against R. trifolii TA1 cross-reacted with strain 31 only in the polar tip region. No polar immunofluorescing tips could be seen with FA against two other strains of R. japonicum or with those against several unrelated microorganisms. Common antigens localized only in a polar region were seen in many rhizobia stained with R. japonicum 31 FA: 22 of 23 strains of R. japonicum, 10 of 17 strains of R. trifolii, 3 of 7 strains of R. melitolii, 3 of 6 strains of R. phaseoli, and 3 of 9 strains of R. leguminosarum had some cells with detectable polar tips. The proportion of R. japonicum 31 cells with polar tips was high throughout the growth cycle. Polar tip staining was not affected by drastic cell treatments. A function was proposed for the polar tip region as a site for attachment. R. japonicum 31 cells attached to each other in a tip-to-tip fashion and endwise to fungal hyphae with the polar tip in contact with the hyphal wall. Binding of fluorescein isothiocyanate-labeled soybean lectin to certain strains of R. japonicum gave additional evidence of polarity. Polar binding of both antibody and lectin may provide insights into relationships between rhizobia and roots of host legumes.

Quantitative analysis of pregnancy-associated plasma proteins in human placenta.

By immunochemical methods and simultaneous measurements of several normal plasma proteins, human placenta was shown to contain elevated quantities of four pregnancy-associated plasma proteins (PAPP's). In the order of increasing amounts, PAPP-A, PAPP-C, PAPP-B, and human chorionic somatomammotropin (PAPP-D) all were present in placenta extracts in quantities greater than could be expected on the basis of their content in maternal blood. In sharp contrast, the placental content of pregnancy zone protein could be entirely accounted for by the maternal plasma present in the placenta. All of the PAPP's appeared to be readily extractible from placental tissue with buffered saline, the large bulk of them being solubilized in the first extraction procedure. However, absorption studies indicated that appreciable quantities of the PAPP's were still present in the insoluble placental residue after 12 sequential extractions with saline. The chorioamniotic membranes were not significantly enriched in any of the PAPP's. Immunochemical analysis of unwashed placental tissue extracts for the PAPP's IgA, and IgM (maternal blood derived), as well as albumin and transferrin (maternal and fetal blood derived), permitted calculations to be made of the amount of blood and PAPP's in placenta. On the basis of these data, it was roughly estimated that a 400-g placenta (wet weight) would occupy 312 ml in volume, and would contain 144 ml of blood. Of this blood, 36 ml would be derived from the mother.

Quantitative fluorescence spectrophotometry of acridine orange-stained unfixed cells. Potential for automated detection of human uterine cancer.

After staining with acridine orange (AO), the nuclei of unfixed cells from the human female genital tract exhibited the same fluorescence behavior previously observed for human and murine leukocytes and mouse ascites tumor cells. With staining conditions chosen to assure saturation of the green-fluorescing AO-nucleic acid complex in normal cells, corrected fluorescence emission spectra were recorded from the entire nucleus of 341 cells taken from 32 normal and 28 abnormal patients. Intensity of the recorded spectra was expressed in phosphor particle units, a fixed arbitrary unit of fluorescence intensity, to display intensity differences among the spectra from the various cell types. In all abnormal samples, one or more cells were found with 530-nm nuclear fluorescence intensity considerably greater than the maximum intensity recorded from normal cells. Determination of the adequacy of 530-nm nuclear fluorescence intensity as a criterion for cancer detection requires additional investigation. Additional criteria, if needed, may be supplied by the metachromasy of AO-stained unfixed cells.

Resistance to tumor growth mediated by Listeria monocytogenes. Destruction of experimental malignant melanoma by LM-activated peritoneal and lymphoid cells.

A murine experimental model of nonspecific tumor destruction mediated by cells activated by Listeria monocytogenes (LM) is described. B16 melanoma growth is prevented or suppressed in the syngeneic host when tumor cells are inoculated in contact with viable LM. In vitro, cultured B16 cells are destroyed by LM immune peritoneal or splenic cells in the presence of the bacterial antigen(s). Activation of LM immune cells in vitro is immunologically specific. Replacement of LM by sheep red blood cells or bovine serum albumin in the in vitro cultures aborts the cytotoxic effect. Further, no tumor cell killing is obtained when thioglycollate-induced or normal peritoneal cells are substituted for LM immune cells in the in vitro cultures. Normal spleen cells in the presence of LM are weakly cytotoxic for B16 cells. Normal peritoneal cells plus LM or LM alone are not. Elimination of thymus derived "T" cells by anti-theta C3H or rabbit anti-mouse brain serum (RAMB) abrogated the cytotoxic effect. Therefore, LM-induced tumor destruction probably occurs through nonspecific mechanism(s) consequent to activation of host "T" cells by specific immune reactivity to LM antigen(s).

SPECIFIC positive and negative selection of rat lymphocytes reactive to strong histocompatibility antigens: activation with alloantigens in vitro and in vivo.

This study compares the functional properties of rat thoracic duct lymphocytes (TDL) after stimulation with strong alloantigens of the major histocompatibility complex (MHC) either in vitro in preparative mixed lymphocyte interactions (MLI) or in vivo in systemic graft-vs-host (GVH) reactions. Comparisons were made of PHA responses and reactivity to the specific priming haplotypes or to third party haplotypes in analytical MLI and in GVH reactions either before or after the activated populations were "parked" in syngenetic T cell-deprived (B) rats. These comparisons can be summarized as follows: 1) TDL populations primed in bulk MLI cultures (MLI-TDL) slowed some evidence of specific positive selection when tested immediately; MLI responses to specific alloantigens were both relatively large and accelerated in tempo, whereas responses to third party alloantigens were diminished but also accelerated in tempo. Specific GVH responses were more marked than in third party recipients but they were also decreased relative to normal, and displayed an abberant dose/response slope. MLI-TDL populations tested after they had been stored in syngeneic B rats showed clear evidence of stable-specific positive selection; specific MLI and GVH responses were enriched relative to third party responses and also in comparison to normal, unselected TDL populations. This finding indicates that GVH and MLI reactivity are probably both functional capacities of the same lymphocyte subpopulation since positive selection by one function (MLI) also enriched for a second (GVH). 2) Parental strain TDL activated in vivo in the systemic GVH reaction in irradiated F1 animals and recovered from the thoracic duct 3 to 4 days later (late GVH-TDL) consisted mainly of blast cells, however, in contrast to MLI-TDL these populations showed no evidence of positive selection when tested before or after parking in B rats. MLI responses to specific alloantigens were minimal, and greatly reduced in magnitude compared to normal. GVH responses to specific haplotypes could be detected, but these were not enriched compared to normal, despite the content in the late GVH-TDL populations of a significant proportion of blast cells presumably activated by host alloantigens. 3) Early collections (less than 40 hr) of parental strain GVH-TDL collected from F1 recipients contained no blast cells and showed impressive degrees of negative selection; they were markedly depleted of both GVH and MLI activity to specific alloantigens but displayed normal reactivity to third party alloantigens. Moreover, specific negative selection was persistent in these populations parked for several weeks in B rats, and indication that a specific subpopulation of reactive cells had been physically eliminated. 4) PHA responses of both MLI- and GVH-activated TDL populations tested either before or after parking in B rats were approximately normal on a per T cell basis...

Effects of sodium periodate modification of lymphocytes on the sensitization and lytic phases of T cell-mediated lympholysis.

Sensitization of mouse splenic lymphocytes in vitro with sodium borohydride, suggesting that the biologic effects of sodium periodate are-treated autologous spleen cells stimulated a one-way mixed lymphocyte reaction and led to the generation of thymus-derived cytotoxic effector cells. These effectors were capable of lysing in 4 hr periodate-treated syngeneic and, to a lesser extent, periodate-treated allogeneic target cells. These results suggest that sensitization by periodate-treated autologous cells could result either from a specific reaction to modified self components or from a nonspecific mitogenic stimulation. Effector cells generated by allogeneic sensitization were detected on periodate-modified targets, irrespective of the H-2 antigens expressed by the targets. The effects of periodate modification on both stimulator and target cells were reversible by sodium periodate are dependent on the formation of a free aldehyde group on cell surface glycoproteins. Pretreatment of stimulator cells with neuroaminidase prevented the effect of periodate treatment, suggesting that the sensitization involves oxidized sialic acid residues. During the 4-hour 51Cr-release assay periodate-treated targets could be used to detect cytotoxic effector cells of any specificity. Fresh spleen cells and lymphocytes cultured for 5 days without antigen or in the presence of lipopolysaccharide did not lyse periodate-treated targets. An increasing level of cytotoxicity was detected on periodate-treated targets when the effector cells were generated, respectively, by stimulation with concanavalin A, by sensitization with periodate-modified autologous cells. Although the lysis of periodate-treated targets is itself nonspecific, effector cell specificity could be determined by selective blocking of the lytic phase with cells syngeneic to the stimulators. These results indicate that a nonspecific interaction can occur between lymphocytes and periodate-treated target cells, but that this interaction leads to lysis only when the lymphocytes were activated to become cytotoxic effectors.

Requirement of precommitted cells as targets for the augmentation of lymphocyte proliferation by leukocyte dialysates.

After our initial report tha leukocyte dialysates containing transfer factor augment the thymidine incorporation of antigen-stimulated lymphocytes, we have adapted the system to microleukocyte cultures. This modification permits both (a) the simultaneous assay of a single dialysate on the cells of multiple individuals, and (b) the assay of multiple dialysates on the cells of a single individual. The data thus secured, demonstrate that dialysates from both skin-test-positive and -negative donors produced similar degrees of augmentation whether the data are expressed as an arithmetic difference or as a ratio. When expressed as an arithmetic difference, the amount of augmentation is increased in proportion to the level of thymidine incorporation of the assay cells when they were stimulated by antigen alone. When expressed as a ratio, however, the degree of augmentation is independent of the response of the assay cells. An analysis of the ability of dialysates to engage previously uncommitted lymphocytes and thus to augment thymidine incorporation, revealed that precommitted cells were required. In these experiments, antigen-reactive cells were deleted from populations of peripheral blood lymphocytes by incubation with purified protein derivative of tuberculin, diphtheria toxoid, or streptokinase-streptodornase in the presence of [3H]thymidine of high specific activity. This deletion depressed or abolished the effect of dialysate on the residual population when it was recultured with the same antigen, but the effect on the response of the remaining lymphocytes to other antigens was unaltered. In this study, leukocyte dialysate appeared to augment nonspecifically the thymidine incorporation of an antigen-specific precommitted clone of lymphocytes. The relationship of these adjuvant effects on peripheral blood lymphocytes in vitro to the specific and nonspecific activities of transfer factor in vivo remains to be elucidated.

The significance of perivascular infiltrations in multiple sclerosis.

143 autopsy cases of multiple sclerosis (19 acute and 124 chronic cases) were analysed histologically for the extent of active demyelination and the degree of infiltration within and outside the demyelinating lesions and in the leptomeninges. The results were compared with the duration of the illness. Infiltrations were found in 60% of all cases but more often (74%) in those with active demyelination. Inflammatory lesions outside demyelinating foci were observed in 27% of the total, and in 80% of them active demyelination was present. Inflammatory lesions in the meninges were present in 41% of the total and in 80% of these were accompanied by active demyelination. The duration of illness correlated with decreasing severity of active demyelination and of perivascular infiltration. Patients treated with cortico-steroids and/or immunosuppressive substances showed no or only moderate inflammatory lesions. The duration of illness in both these groups was significantly longer than the average of untreated patients. The significance of these pathological findings for the CSF cytology in multiple sclerosis is discussed.

Fast axoplasmic transport in the fibres of chromatolysed neurones.

1. The rate of fast axoplasmic transport in cat sensory nerves was determined in sciatic nerves above transections made low in the popliteal fossa some 6-165 days beforehand. The pattern and rate of movement of the crest of labelled components in the nerve fibres after injecting the L7 dorsal root with [3H]leucine was used to characterize fast axoplasmic transport. 2. The mean rate and S.D. found on the transected side was 424 + 33 mm/day compared with 432 +/- 34 mm/day for the control nerves. These rates were not significantly different and were similar to the rate of axoplasmic transport previously reported to be 410 +/- 50 mm/day. The results gave little support for the hypothesis that a speeding up of the rate of fast axoplasmic transport is the signal for the initiation of chromatolysis. 3. The amount of transport shown by the level of activity in the crests on the chromatolytic and control sides relative to the "pool" of radioactive materials remaining in the cell bodies of the ganglion were also similar. The significance of these findings was discussed with respect to changes in the cell bodies known to take place during chromatolysis and the stability of the axoplasmic transport mechanism in nerve fibres.

Tumor-associated antigens in human myeloma.

Antiserum was generated in rabbits to the RPMI 8226 tissue culture line of human myeloma cells, and its reactions with fixed smears of bone marrow aspirates from patients with multiple myeloma, macroglobulinemia, benign monoclonal gammopathy (BMG), leukemia, and nonneoplastic plasmacyosis was assessed by indirect immunofluorescence. After absorption with preparations of bone marrow from normal individuals, the antiserum reacted to a significantly higher titer with a specific subpopulation of plasma cells in smears from 81% of patients having multiple myeloma and 50% of patients having BMG than with cells in smears of bone marrow aspirates from normal individuals or patients having leukemia or nonneoplastic plasmacytosis, or than with cells in smears of peripheral blood from patients having Hodgkin's and non-Hodgkin's lymphoma. Absorption of the antiserum with RPMI 8226 cells or with a bone marrow preparation from a patient with multiple myeloma but not the Jijoye line of Burkitt's lymphoma reduced reactivity for cells in myeloma bone marrow. The antiserum reacted at a lower titer with the Jijoye and EB-3 lines of Burkitt's lymphoma, the RPMI 4098 cell line of normal human lymphocytes, and culture lines of human melanoma and osteogenic sarcoma than with the RPMI 8226 cells or bone marrow from certain patients having multiple myeloma. Approximately 50% of the cells reactive with antiserum to RPMI 8226 cells in the bone marrow of patients with multiple myeloma were not producing immunoglobulin, as assessed by double immunofluorescence assay. The data suggested that a subpopulation of plasma cells in the bone marrow of patients with multiple myeloma possesses a tumor-associated antigen.

Purification and serological characterization of the major envelope glycoprotein from AKR murine leukemia virus and its reactivity with autogenous immune sera from mice.

The major envelope glycoprotein (gp71) from AKR murine leukemia virus (MuLV) was purified and its serological reactivity with heterologous and autogenous immune mouse sera was examined. Homologous and interspecies competition radioimmunoassays using antisera to Rauscher-MulV gp69/71 or Friend-MuLV gp71 or antisera to feline leukemia virus to precipitate 125I-labeled gp71 from various MuLV showed that distinct differences exist between Rauscher- or Friend-MuLV and AKR-MuLV glycoproteins. Characteristically the AKR-MuLV gp71, in contrast to FLV or RLV gp71, does not compete fully in homologous or interspecies radioimmunoassays with iodinated Friend of Rauscher glycoproteins. Purified 125I-labeled AKR-MuLV gp71, in contrast to the Rauscher- or Friend-MuLV glycoproteins, reacts with normal (autogenous immune) mouse sera in direct radioimmune precipitation assays. Competition experiments further demonstrate that this is a predominant immunological reactivity of normal mouse sera which had previously been detected by radioimmune precipitation assay against intact virions.

Relationships between intracisternal type A and extracellular oncornavirus-like particles produced in murine MOPC-460 myeloma cells.

Oncornavirus-like particles of the "A" (both intracisternal and intracytoplasmic) and "B" or "C" (extracellular) types are produced by murine MOPC-460 myeloma cells. This communication describes a comparative study on tracisternal A and extracellular particles. Both types of particles contain an RNA-dependent DNA polymerase activity, traces of 35S and 70 S RNA in addition to larger amounts of degraded RNA, and proteins of approximately 76,000 and 45, 000 daltons. The 76,000-dalton proteins from intracisternal A and extracellular particles have the same cyanogen bromide peptides. Hybridization kinetic analysis indicates that the RNAs in the two particles are identical or very closely related and share partial homology with Moloney leukemia virus RNA. In contrast, the particles appear to have little or no relationship to murine mammary tumor virus as judged by several different criteria. Electron microscope studies indicate that the extracellular particles arise from the budding of core components through the plasma membrane. These results suggest that the intracisternal A and extracellular oncornavirus-like particles produced by MOPC-460 cells are closely related.

Prediction of azathioprine intolerance in transplant patients.

One cause of transplant rejection is curtailment of immunosuppressive therapy due to leucopenia. To determine those patients most apt to develop leucopenia due to azathiprine the granulocyte response to intravenous injections (i.v.) of hydrocortisone was evaluated in 10 patients who had rejected their grafts at least six month previously. 5 patients who had rejected their grafts with concomitant severe leucopenia had an inadequate response to hydrocortisone, while in the other 5, who had tolerated the drug, the response was similar to that of normal controls. Based on these observations, all the transplant candidates underwent the hydrocortisone stimulation test the results of which were correlated with their subsequent clinical course. All medical decisions were based on events other than the steroid test. 8 leucopenic patients underwent splenectomy. 6 improved their granulocyte response to hydrocortisone and tolerated azathioprine after transplantation. 2 patients who underwent splenectomy and an unoperated leucopenic man were unresponsive to the hydrocortisone test, did not tolerate azatioprine after transplantation and rejected their grafts. 4 candidates with normal responses to i.v. hydrocortisone received transplants uneventfully. In all 13 patients transplanted since the beginning of this study, the hydrocortisone test correctly predicted their tolerance to azathioprine.

Alterations of alpha2-globulin and the clinical response in patients with prostatic cancer following cryotherapy.

Evaluation of alterations in the level of alpha2-globulin in the serum of 18 patients with prostatic cancer prior to and following cryotherapy of their primary prostatic tumor and the clinical response of these patients disclosed: (1) a progressive increase in the level of alpha2-globulin and the incidence of patients with significantly elevated levels of alpha2-globulin, i.e., greater than or equal to 1.30 g/100 ml, with a progression of the stage of their malignancy; (2) a decrease in the levels of alpha2-globulin in the serum of 14 of 18 (78%) patients following cryotherapy, and (3) a favorable clinical response in 11 of 14 (79%) patients with prostatic cancer showing a decrease in alpha2-globulin following cryotherapy. While limited to the study of a relatively small patient population, the present results suggest a prognostic potential for alpha2-globulin, particularly as applied to stage identification in prostatic cancer. Pending confirmation by evaluation of a larger patient population, it may even provide objective criteria for monitoring the clinical response of an individual following cryotherapy of the prostate.

The safety of dexamethasone and hydroxyethyl starch in the multiply leukapheresed donor.

A total of 23 leukaphereses were performed on five normal, healthy donors for the purpose of providing granulocyte transfusions to septic leukemia patients with granulocytopenia. Dexamethasone 7.25 to 7.50 mg was given orally 10 to 12 hours prior to each donation, and an average of 304 ml of hydroxyethyl starch (HES) was given intravenously during each procedure. During the period of observation for each donor, there was no significant change of total leukocyte and platelet counts, total bilirubin, alkaline phosphatse, LDH, SGOT, creatinine, BUN, and uric acid determinations. Changes in the concentrations of serum protein, albumin, cholesterol, and glucose were thought to be due to hemodilution. Partial thromboplastin and prothrombin times remained within normal limits following collection procedures. Hemoglobin levels decreased transiently following the first three leukaphereses in all donors, but fell progressively to 11.8 gm/dl in one donor undergoing seven procedures in a 35-day period. Dexamethasone and HES in these doses can be given safely to multiply leukapheresed donors.