Rapid diagnosis of thyroid disease using carbonic-anhydrase immunoassay. Erythrocyte carbonic-anhydrase-I (C.A.I.) concentration was measured by radial immunodiffusion in 30 thyrotoxic patients and 15 myxoedemic patients, the results being compared with those found in a control group of 90 euthyroid individuals. In thyrotoxicosis the erythrocyte-C.A.I. concentration was found to be below 10-29 mg/g haemoglobin, whereas the normal range varied from 11-1 to 22-5 mg C.A.I./g Hb. Low values were found in 4 patients with triiodothyronine(T3)-toxicosis. In myxoedema, the mean erythrocyte-C.A.I. concentration, though elevated at 20-37 mg/g Hb, fell within the normal range. In pregnancy, the erythrocyte-C.A.I. concentration rose with time, the mean value at tern being 20-6 +/-S.D. 3-3 mg/g Hb. In a group of women taking oral contraceptives, a low erythrocyte-C.A.I. concentration was noted--an effect which was most obvious among those taking low-oestrogen contraceptives. It is suggested that erythrocyte-C.A.I. estimation provdes a rapid screening test for thyrotoxicosis.


Antiviral activity in milk of possible clinical importance. In human and in cow's milk an antiviral activity has been detected which does not seem to be related to antibodies or other known virus inhibitors. The antiviral activity lay in a relatively heat-stable macromolecule belonging to the non-fatty part of milk.


Coronary risk factors and socioeconomic status. The Oslo study. Coronary heart-disease (C.H.D.) had been reviewed as a "manager's disease". However, deaths from C.H.D. are now said to be more common in groups from the lower social classes than in those of higher socioeconomic status. We have examined wheather these differences in C.H.D. mortality can be explained by differences in the conventional risk factors for C.H.D.


Transnasal transsphenoidal approach to the sella. The transnasal transsphenoidal approach to the sella turcica has been refined and perfected through advances in medicine and technology. Originally performed for excision of pituitary adenomas, its present primary use is pituitary ablation is hormonally dependent diseases. Our study of 55 patients who have undergone this procedure during the years 1970-1975 demonstrates that this microscopic technique gives excellent exposure and visualization of intrasellar contents. Our method gives excellent exposure and visualization of intrasellar contents. Our method with several instrument modifications is discussed. It is technically an easy procedure with a minimum of morbidigy and mortality.


Phosphotungstic acid-iron-haematoxylin staining method for osteoid, boundary bone and bone components in paraffin sections. A new staining technique which stains osteoid and bone tissue differentially and also demonstrates boundary bone, pathological osteoid and the changes in ageing, pathological and dead bone matrix in decalcified paraffin or low-viscosity-nitrocellulose bone sections was developed. This phosphotungstic acid-iron-haematoxylin (PTAIH) method is based on pretreating the sections with phosphotungstic acid followed by an iron alum mordant and staining in haematoxylin with subsequent timed differentiation, at certain stages of which the features listed above appear. Van Gieson's picrofuchsin is then used as a counterstain. After standard differentiation osteoid appears red in sharp contrast with the black bone, young and woven bone, old and lamellar bone, and allows one to demonstrate changes in stainability of diseased osteoid and bone matrix, and dead bone. With the differentiation done individually and interrupted at certain stages it is possible to distinguish between various bone components depending on the amount and quality of their in vivo mineralisation. Comparison with controls showed that in this respect the method is more sensitive than the curremt staining techniques of undecalcified bone sections since it demonstrates not only unmineralised and fully mineralised tissues but also shows the poorly calcified, demineralised and ill-calcified bone components. The advantages of the method compared with those using undecalcified sections are its simplicity, suitability for fixed and decalcified material in any unspecialised histological laboratory and the fact that osteoid and other bone components can be studied in sections of unlimited size and in undisturbed relationship to their surrounding soft tissues.


Methods of fixing, sectioning and staining amphibian eggs for cytological study. A combination of methods for fixation (sublimate, cobalt nitrate, formaldehyde, acetic acid in water), inclusion (celloidin dissolved in methyl salicylate, paraffin-paraplast) and staining was used to make serial sections easy, to avoid clefts and to give a good picture of segmentation mitoses, as well as a good contrast of yolk and cytoplasmic components. Four methods of staining were used concerning the Urodele eggs: Safranin-methyl blue-orange G, safranin-picro-blue black naphthol (Curtis), safranin-violet crystal-orange G (Flemming) and Feulgen-methyl blue-orange G. The achromatic apparatus of the normal segmentation mitoses is clearly delineated and the relationships between astral fibers and yolk are different at metaphase and anaphase. By these methods, particularly suitable for demonstration of nuclei, cytoplasm and achromatic apparatus, the cleaving egg may be used as a test for the inhibition of achromatic apparatus and chromosome damage by antimitotic substances. The contrast between vitelline cytoplasm and cytoplasmic non-vitelline abnormal fibrillar systems, produced by transformation of astral and diastematic fibres, is made particularly evident by these methods of staining.


A lecture on lecturing.

A lecture on lecturing. There are major differences between a lecture and a paper for publication. Often the printed word is spoken at meetings, a kind of compulsive public reading which has robbed the lecturer of the chance of oratory and the audience of a little enjoyment. The simple fact is that although doctors read aloud badly (actors do this far better) most can learn to speak spontaneolsly and with animation; but this requires time and effort, both of which are donated in a miserly way. The successful lecturer is generous and considerate of his audience--a rare being at medical meetings.


Evaluation of an integrated multi-media learning system. Two teaching methods were compared: (1) the traditional way with textbooks, practical guidance, personal demonstration, and (2) an integrated multimedia system with text programme, slide-sound-show accompanying the experiment, and lecturer. Two samples of thirty-one students each were classified in twin groups by means of psychological tests. Both groups worked with the identical pretest/post-test and different training procedures. The integrated multi-media system produced highly significant more gain in the theoretical sphere, greater personal confidence, and lower experimental error. Furthermore positive attitude of the students towards the experiment was demonstrated. The improved performance was statistically proved by t-test and covariance analysis.


A method of teaching epidemiology in the clinical setting. It is important to provide an epidemiological presence when teaching clinical medicine to undergraduate medical students in order to supply the underlying facts about disease. A series of pictorial or tabular presentations which together make up a 'master set' of Data Boards has been designed to give the medical student a visual picture of epidemiology. Each Data Board is constructed in such a way that it can be readily interpreted by a medical student at the bedside or in similar situations to those where he is expected to interpret clinical signs and symptoms of disease. The present use of Data Boards in clinical teaching is described and suggestions for future applications are made.


[Pathological-anatomical findings in 70-102 years old Caucasians (author's transl)]. The macroscopic findings of 2385 autopsy protocolls of 70-102 year-old males and females were analysed for frequency and functional significance of diseases. 1. Severe coronary arteriosclerosis was found up to 60% of the cases, with increasing frequency in higher age groups. However decrease of acute myocardial necrosis was stated including the highest age group. 2. The mean values of heart weights decrease in the highest age groups. 3. While the frequency of arteriosclerosis in the cerebral arteries increases the frequency of ischemic lesions of the brain decreases. 4. Benign hyperplasia of the prostate is found in 85% of the cases. 5. 80% of the patients had severe substantial chronic pulmonary emphysema and acute tracheobronchitis. These pulmonary lesions are supposed to be the functionally most important disease in patients of old age.


Electron-microscopic identification of infectious particles of lymphocytic choriomeningitis. LCM virus, strain WE--grown on L cells--and labeled with 3H-uridine was centrifuged to equilibrium in a sucrose density gradient and examined in fractions for infectivity, incorporated radioactivity, and electron-microscopic features. The peak of infectivity is congruent with the one of radioactivity (density = 1.17 g/ml). LCM virus specificity of the radioactive peak was proved by precipitation of the radioactivity with anti-LCM virus antiserum. The peak fractions showed an abundance of 106 +/- 14 nm (1s) particles. They could be agglutinated with specific anti-LCM virus antiserum but not with antiserum directed against the histocompatibility (H-2) antigens of L cells.


Serum protein electrophoresis (SPE) and immunoglobulin levels in breast carcinoma. Serum protein electrophoresis (SPE) and immunoglobulin levels were studied in 50 patients with breast carcinoma before therapy (17 with stage I disease; 18 stage II; 15 stage III or IV), and in 55 patients after primary lesions were surgically removed and gave no evidence of recurrence. The most consistent and statistically significant abnormalities observed on SPE were those of albumin, globulin, and alpha-2 globulin. Patients who were free of disease during follow-up examinations had higher albumin and lower globulin and alpha-2 globulin levels than those with advanced breast cancer (P = 0.003, P = 0.03, and P = 0.001 respectively). There was no difference in concentrations of immunoglobulin G, A, or M among the 5 groups. Routine laboratory tests of hemoglobulin, lymphocyte count, calcium, alkaline phosphatase, creatinine phosphokinase, lactic dehydrogenase, and transaminase also did not differ among the groups, but carcinoembryonic antigen was significantly elevated in patients with stage III and IV disease.


The pathogenesis of amyloid deposition: a new hypothesis. Amyloid proteins are probably derived from a variety of precursor glycoproteins. It is postulated that there may be at least two key events in the pathogenesisi of amyloidosis. The first is an increase in the load of glycoprotein being brought to a site of degradation. In the case of myeloma this might be in the form of excess immunoglobulin light chains. In the case of secondary amyloidosis the form taken could be enzyme-alpha-globulin complexes. The second is an inability of the degrading site to handle the arriving substrate at a sufficiently rapid rate, the rate limiting step being at some point along the degradation pathway. We postulate that an acquired enzyme deficiency prevents removal of the carbohydrate moiety of the presented glycoprotein. This results in the accumulation of a normal intermediate (amyloid protein) during the breakdown of the glycoprotein substrate. Evidence for the operation of these mechanisms is discussed and their detailed nature and implications considered.


Tissue specificity of nonhistone proteins from human chromatin. Nonhistone proteins were isolated from human placental and tonsillar chromatins. Antiserum was prepared against a complex from some nonhistone proteins and DNA (NP-DNA) from placental chromatin. With the help of polyacrylamide gel electrophoresis and immunological methods the tissue specificity of human chromatin nonhistone proteins was established. The described organ immunogenic specificity of the complex of DNA and nonhistone protein (NP-DNA) from human chromatin is in accordance with data published on similar complexes from different animal organs. Besides, it is shown that shearing of chromatin leads to large chifts in NP-DNA concentrations required for maximum complement fixation in the presence of the prepared antiserum. This may probably be due to a damage of certain chromatin super structures which involve some of the nonhistone proteins and DNA sequences from both the more condensed and less condensed parts of chromatin.


Biochemical and electron microscopical evidence for the presence of oncorna viruses in spleen tissue from two patients with haematological malignancies. In extracts of spleen tissue from two patients with haemotological malignancies an RNA dependent DNA polymerase was found in particles with a density of 1.16, that is at the density of oncorna viruses. After treatment with noniomic detergents the enzyme activity was found in particles with a density of 1.23-1.24, similar to the density of oncorna viral cores. A simultaneous detection test with this core fraction material for 70 S RNA and RNA dependent DNA polymerase was positive for both patients. Electron microscopical inspection of the material with a density of 1.16 revealed immature C-type virus like particles, various stages of maturing particles and a number of particles resembling mature C-type oncorna viruses. In two normal spleens from patients with carcinoma of the colon and oesophagus respectively and in three spleens from patients with no history of malignancy no RNA dependent DNA polymerase was found. Material from one normal spleen was examined in the electron microscope and no virus-like particles were seen.


Effect of a digitalis drug on ventricular premature beats. To determine the efficacy of digitalis drugs in suppressing ventricular ectopic activity, 142 patients with frequent (greater than 1 per minute) ventricular premature beats underwent acetyl strophanthidin tolerance testing. In 65 patients (46 per cent), frequency and grade were reduced during testing. In 37 (26 per cent), the ectopic activity remained unaltered; frequency increased during testing in the remaining 40 patients (28 per cent). In the group with a suppressive effect, ventricular premature beats decreased by 82 per cent, with complete abolition of arrhythmias in 46 per cent. The three groups were not distinguishable clinically by either the type or the extent of heart disease. The antiarrhythmic action of acetyl strophanthidin did not appear to depend upon its positive inotropic action. In some patients it appears to be due to an indirect reduction of Purkinje-fiber automaticity resulting from enhanced vagus-nerve activity that thereby lessens adrenergic tone on the heart.


Generation of an esterolytic and kinin-forming kallikrein-alpha2-macroglobulin complex in human serum by treatment with acetone. Kinin-forming and esterolytic activity in human citrate plasma has been activated by treatment of the plasma with acetone. By far most of the esterolytic activity if not all of it was recovered in an alpha2-macroglobulin (alpha2M) kallikrein complex (SI) which was characterized and purified by chromatography. Little if any esterolytic activity was present which could be ascribed to free plasma kallikrein. The alpha2M-kallikrein complex had kinin-releasing activity though much less than free plasma kallikrein, relative to esterolytic potency. This explains that a considerable fraction of the kinin-forming potential of acetone-activated plasma resides in free plasma kallikrein although it represents only a very small portion of the total kallikrein store. Like free plasma kallikrein the alpha2M complex releases kinin from LMW-kininogen less efficiently than from HMW, in systems of purified components. In whole plasma, the efficiencies change: whereas plasma kallikrein is rapidly inactivated by endogenous inhibitors, the alpha2M complex is protected from further inactivation and capable of releasing kinin continuously if slowly, attacking also LMW-kininogen after HMW-kininogen has been consumed by free kallikrein. While the alpha2M-complex in this respect differs functionally from free plasma kallikrein and explains earlier observations suggesting the presence of two kininogenases, it seems doubtful now that two truly different kininogenases exist in human plasma. The results suggest that acetone predominantly inactivates full inhibitors of kallikrein such as C1INH whereas alpha2M is somewhat more resistant and (pre-)-kallikrein even more. Depending on the time and temperature of acetone treatment one obtains more or less total kallikrein and varying proportions of free to bound enzymes. It is likely that acetone does not turly trigger an activation of prekallikrein but supports spontaneous activation by slowing down the control of the feedback reinforcement of this activation, by damaging inhibitors.


Amniotic fluid tests for fetal maturity in normal and abnormal pregnancies. The lecithin-sphingomyelin (L/S) ratio is acknowledged to be superior to most procedures for predictinf fetal lung maturity in normal pregnancy. In complicated gestations, however, errors have been reported. This study involves 686 normal gestations and 389 pregnancies complicated by fetomaternal diseases. The L/S ratio, creatinine level, and percent of fat-staining cells were measured in samples of amniotic fluid from these patients. The results showed good correlation of all three tests with fetal maturity as measured by weight, Dubowitz criteria, and incidence of respiratory distress syndrome in the normal patients. In the complicated pregnancies, however, the creatinine was unacceptable in up to 30% of the cases. The L/S ratio likewise decreased in accuracy for all parameters of fetal maturity measured. The Nile blue staining of the fetal cells appeared to be the most consistent technic in these cases. A fetal maturity battery comprised of these three assays and other methods of assessing fetal health is advocated in pregnancies complicated by certain disease states.


Amniography for the early detection of neural tube defects. Among the methods available for the midtrimester diagnosis of neural tube defects, it has been suggested that amniography be used to determine the presence of meningomyelocele. Twelve studies are reported in which the procedure was utilized. In 3 cases, a neural tube defect was present but was not detected by this method. The experience reported here challenges the value of this procedure and suggests that it should be evaluated further by utilizing it in cases where a positive diagnosis has been made by other methods and pregnancy termination is to be performed.


Nile blue and fetal maturity. Further investigations. The consistency of results using Nile blue staining of fetal cells in amniotic fluid to estimate fetal maturity has been demonstrated. Recently, evidence to support this method has been published from this laboratory. Several biochemical and technical factors were cited as important to the success of the method. This report deals with further testing of several Nile blue dyes (hydrochloride and sulfate) necessitated by changes in production and federal regulation of this compound. The results show Nile blue hydrochloride (HCl) to be the most superior dye regardless of color index. Buffered solutions (6.6) of Nile blue A (sulfate) improve its performance, but not to levels demonstrated by the HCl preparation. Storage at room temperature adversely affects each dye; however, the sulfate variety appears to be the most unreliable under these circumstances. For best results, it is recommended that Nile blue HCl be used and the solution (buffered or normal) be kept refrigerated.


Phytohemagglutin-induced lymphocyte transformation in oral contraceptive users. Phytohemagglutin (PHA)-induced lymphocyte transformation (PILT) was determined in 217 women taking oral contraceptives and 203 control women by means of the uptake of 3H-thymidine into DNA of lymphocytes cultured in heterologous serum. Depressed PILT responses were observed in oral contraceptive users as compared with age-matched controls, and the magnitude of depression correlated with the duration of oral contraception and was inversely related to the clinical progestagenic potency of the component steroids. An additional group of 21 women, tested within 1 year (mean 3 months) of cessation of oral contraception, showed persistent depression of PILT responses. Suppression of lymphocyte transformation in autologous as compared with homologous, normal serum suggests that serum inhibitory factors amy be important. We found no evidence for a direct suppressive in vitro effect of synthetic estrogens and gestagens. The prevalence of autoantibodies in oral contraceptive users was similar to that in control subjects.


[New experiences with bleomycin].

[New experiences with bleomycin]. 26 patients all with squamous call carcinoma of the head and neck were treated primarily with Bleomycin (maximal dose 300 mg by i.v. injections of 15 mg every second day). In 10 of the patients a significant regression of the tumor was achieved. In one particular case no histological evidence of tumor could be found. The expected side effects of Bleomycin were observed. Two patients died of heart-lung failure. Histological examinations showed signs of cytostatic pneumopathy.


Tumoral drugs as possible blastogenic agents the problem of anti-blastic medication. The author studies the possible transplacentary carcinogenic drugs used in human therapeutics; the study is of an experimental nature, the pregnant mother rat being injected at the 20-21st days of gestation with double the kg/day dose used in human therapeutics. A total of 1,264 rats (Wistar) were utilized, careful note being made of the possible appearance of tumors throughout the lives of the animals. The products and tumoral percentages, both benign as well as malignant, were as follows: Oncotiotepa (0%); Daunoblastine (3.3%); Vinblastine (9%); Bleomycine (12.8%); 5-Fluoro-Uracil (12.87%); Lyovac (cosmogen) (16.6%); Genoxal (17.14%); and Natulan (37.42%). The benign forms predominated in all the tumors produced, but with some of the drugs the malignant varieties produced were made as 39.3% of the tumors. The location and type of tumors were variable; there being cutaneous, glandular, mammary, hepatic, renal, and tumors of the nervous system; there were also tumors of epithelial, connective and nervous variety.


Silver deposition in mouse glomeruli.

Silver deposition in mouse glomeruli. Administration of 6 mM silver nitrate in the drinking water of mice resulted in deposition within the glomerular basement membrane of silver granules which were detected on electron microscopy after 12 days. Larger aggregates were detected in the basement membrane and mesangium when silver ingestion was extended to 14 weeks. The silver deposits did not alter significantly over a period of 21 weeks after silver nitrate ingestion was stopped.


Morphological and immunological evidence of coagulopathy in renal complications of pregnancy. Renal biopsies in 14 patients with P.E.T. or eclampsia showed constant I.F. reactions for IgM and fibrin, with frequent reactions for C1q and C3. The glomeruli showed reversible mesangial proliferation and swelling, with characterictic E.M. deposits, and segmental lesions were present in seven patients. Similar I.F. reactions occurred in three other patients with clinical diagnoses of P.E.T. whose biopsies demonstrated coexistent glomerular disease. Serum complement studies showed a significant rise in C3 in the third trimester of normal pregnancies and a further significant elevation in C1q and C3 in the third trimester of a series of unselected P.E.T. patients. In contrast, four patients from the biopsy series with eclampsia or severe P.E.T. showed profound depression of serum C3 and C4, at the time of maximum clinical severity, which was shown to return to normal in two patients. The I.F. findings confirm those of Petrucco et al (6), and, with the other data, suggest that immune-complex deposition and activation of the classical complement pathway could interrect with intravascular coagulation to produce the glomerular lesions of P.E.T. and eclampsia.


Lipofuscin in neuronal aging and diseases. The increasing intraneuronal accumulation of lipofuscin has been linked to the aging process by a striking linear correlation between the degree of accumulation and chronological age. It has been established that age pigments are soluble in polar and nonpolar solvents; the pigment is autofluorescent and stains with PAS, Sudan black B, Nile blue osmic acid and ferric ferricyanide techniques. Whether all pigments exhibiting these properties are identical, or at least closely related, regardless of the surrounding tissue, animal species and age of the individual, is debatable. Pigment formation has been demonstrated in young and aged animals as well as in individual subjected to experimental stress and to dietetic and environmental interference. Electron microscopic studies in animals have shown a considerable variability in the fine structure of individual lipofuscin granules but the presence of "lucent vacuoles" surrounded by a unit membrane is one of the characteristic features of neuronal lipofuscin in the aged. Recently, electron microscopy, utilizing the freeze-etching technique, has provided convincing evidence which disproves the earlier view that lucent vacuoles are the remnant of lipid material dissolved and removed during the preparation of the tissues for microscopic examination. These vacuoles have also been demonstrated in freshly frozen material not previously fixed or immersed. Vacuolated pigment granules occur earlier in the area postrema than in other regions of the rat brain (Hasan and Heyder 1974). Regional differences in the time sequence of pigment deposition are present.


Blood groups changes in preleukemic states. Modifications of blood groups in the course of malignant hemopathies are related to the disease itself and appear to be essentially clonal. They refer not only to the glycolipidic ABH and associated antigens, but also to other blood group systems or other genetic markers. These multiple abnormalities are observed in the preleukemic states as well as in the actual leukaemias. Similar but limited to one genetic unit abnormalities have been noticed among apparently normal subjects, especially in the aged. The various modifications observed are indeed related to the genetic function, and the clonal character is in favor of this assumption. The evidence of abnormalities affecting two genetic systems which are linked ABO and AK is a further argument supporting this hypothesis. Furthermore, the distribution of immunoglobulin marker abnormalities corresponds to that of one of the haplotypes of the Gm genotype. Finally, we were able to demonstrate that the glycosyltransferase itself is abnormal too, in the case of ABH antigen abnormalities. In all the cases investigated to date, when apparently new characters seemed to be observed, they are in fact unconverted substrates, which are to be compared to the carcinoembryonic antigens.


Problems relating to immunoselection of leukemias. Problems relating to immunoselection of neoplastic, in particular leukemic, cell lines are reviewed. Since there is ample evidence that specific immune reactions of the host against malignant neoplastic cells do occur, it becomes important to consider the effectiveness and the relevance of immunity in suppression or elimination of neoplastic growth. Emphasis is placed on experimental results obtained in syngeneic tumor-host combinations, because they more closely resemble the situation of spontaneous tumorigenesis or leukemogenesis than xenogeneic or allogeneic model systems. Studies of types of neoplasia observed in cases of human immunodeficiency syndromes offer an important insight into problems involved in immunoselection of leukemic cell lines: the marked predominance of leukemias and lymphoreticular neoplasias in immunodeficient patients invites speculation on both the mechanisms of leukemogenesis and the relative importance of the immune system in eliminating malignant neoplastic cells.


Binding of triton X-100 to diphtheria toxin, crossreacting material 45, and their fragments. Binding of the nonionic detergent [3H]Triton X-100 by diphtheria toxin, by the nontoxic serologically related protein crossreacting material (CRM) 45, and by their respective A and B fragments has been studied. If first denatured in 0.1% sodium dodecyl sulfate, all of the proteins with the exception of fragment A bind increasing amounts of Triton X-100, reaching a maximum of more than 40 mol bound per mol of protein when the detergent concentration exceeds its critical micelle concentration. No measurable amount of Triton X-100 is bound by native toxin or its A fragment of any concentration of the detergent. Undenatured CRM45 or its B45 fragment, on the other hand, readily became inserted into Triton X-100 micelles when the detergent reaches its critical micelle concentration. The results show that the toxin molecule contains a hydrophobic domain located on the portion of the B fragment that is linked to A. This region is masked in native toxin. Based on these findings, a model is proposed to describe how fragment B facilitates the transport of the enzymically active hydrophilic fragment A across the plasma membrane to reach the cytoplasm.


Analysis of HeLa cell hypoxanthine phosphoribosyltransferase mutants and revertants by two-dimensional polyacrylamide gel electrophoresis: evidence for silent gene activation. The spot corresponding to hypoxanthine phosphoribosyltransferase (HPRT; IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) has been identified in two-dimensional polyacrylamide gels of HeLa cell extracts. This spot is absent in gels of 24 HPRT dificient mutants. A missense mutant displays a new HPRT spot at the same molecular weight but different isoelectric focusing position. Five independently isolated revertants of the missense mutant display spots corresponding to both the wild-type and mutant proteins indicating that they synthesize HPRT from two separate genes. If the missense protein is synthesized from a mutated form of the initially active HPRT gene, then wild-type HPRT protein in the revertants must be snythesized from a newly activated but prevously silent wild-type gene. The newly activated gene in the revertants of the missense mutation appears unstable producing a high frequency of spontaneous HPRT mutants.


Bvr-1, a restriction locus of a type C RNA virus in the feline cellular genome: identification, location, and phenotypic characterization in cat X mouse somatic cell hybrids. Somatic cell hybrids were constructed between BALB/c-RAG mouse cells and feline lymphoma cells by the hypoxanthine-aminopterin-thymidine selection scheme. RAG cells spontaneously produce an endogenous B-tropic type C virus. Cat-mouse hybrids preferentially segregate feline chromosomes and retain murine chromosomes-demonstrable by karyotypic and isozyme analyses. Despite the presence of the complete mouse genome, including the viral genome, virus production was diminished to 1-5% of the levels observed in RAG parents based upon particle-associated RNA-dependent DNA polymerase (reverse transcriptase) activity in the culture fluid. Thirty-seven hybrids made on four different occasions had suppressed virus levels, and no hybrids expressed parental virus levels. Reverse selection experiments on 6-thioguanine demonstrated that a restriction gene, tentatively named Bvr-1, was linked to the feline structural genes for hypoxanthine phosphoribosyltransferase (IMP:pyrophosphate phosphoribosyltransferase; EC 2.4.4.8) and glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ 1-oxidoreductase; EC 1.1.1.49) in cats, probably on the X-chromosome. The genetic mode of action of Bvr-1 is trans dominant in restriction of murine leukemia virus. The restriction locus results in a block late in virus maturation but prior to release, since expression of antigens for viral structural proteins and matrue budding particles is apparent on surfaces of restriced hybrid cells but not in high-speed pellets from culture fluid of restricted cells.


Antiviral effect on MS-2 coliphage obtained with a synthetic antigen. The coat protein of bacteriophage MS-2 was cleaved with cyanogen bromide to yield three fragments, possessing the sequence 1-88 (P1), 89-108 (P2), and 109-129 (P3), respectively. The mixture of peptides P2 and P3, which could not be separated, was found capable of inhibiting the neutralization of the phage by antiserum to the whole MS-2. The peptides corresponding to P2 and P3 were therefore synthesized. The synthetic P3 had no capacity to interfere with neutralization of MS-2, not did its macromolecular conjugate with multichain poly(DL-alanine) elicit neutralizing antibodies. On the other hand, the synthetic P2 was very efficient in inhibiting the inactivation of the phage by the antiserum against phage. Furthermore, a synthetic antigen prepared by attachment of P2 to multichain poly(alanine) incuded antiserum in rabbits that was capable of neutralizing MS-2 activity almost as efficiently as the antiserum prepared against the intact coat protein. This inactivation is specific, because it can, in turn, be totally inhibited by P2 peptide.


Immunological crossreaction between alpha-fetoprotein and albumin. Alpha-fetoprotein (AFP), a fetal protein associated with certain tumors, and serum albumin did not corssreact immunologically in native form. Unfolding of their polypeptide chains by reduction of the disulfide bonds followed by carboxamidomethylation produced derivatives with immunochemical properties different from those of the native proteins. Precipitating reactions and radioimmunoassays preformed with antibodies to such unfolded derivatives showed strong crossreactions between albumin and AFP. These assays were found to lack species specificity: AFPs and albumins from different species were equally active. Several other proteins unrelated to AFP or albumin did not react or reacted much less in these assays. These results support the conclusions derived from recent sequence data that AFP and albumin are structurally related and have a common ancestral gene.


Human alpha-fetoprotein as a modulator of human lymphocyte transformation: correlation of biological potency with electrophoretic variants. Human alpha-fetoprotein (HAFP) isolated by immunoadsorbent column was shown to suppress the mitogenic response of human lymphocytes to phytomitogens, antihuman thymocyte antiserum, and the mixed lymphocyte culture. HAFP isolated from the sera and ascitic fluid of five hepatoma patients, and from fetal liver, varied in biological potency over three orders of magnitude. Extended agarose gel electrophoresis and crossed immunoelectrophoresis demonstrated three molecular species of HAFP. Quantitation of the three species revealed a correlation between the relative amount of the most negatively charged species and biological potency. Treatment of HAFP with neuraminidase to remove completely sialic acid residues did not alter the biological potency, but converted the three species to two species having slower electrophoretic mobilities. We conclude that differences in sialic acid content are only partly responsible for the microheterogeneity demonstrated by HAFP, and that variability in another charged moiety is also present. Variation in the relative proportions of the different molecular species of HAFP may be important in the regulation of its immunosuppressive properties.


Simultaneous detection of reverse transcriptase and high molecular weight RNA in tissue of patients with Hodgkin's disease and patients with leukemia. Complexes of high-molecular-weight RNA and reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) have been detected in 14(77.8%) of 18 spleen from patients with Hodgkin's disease and in all samples tested of peripheral leukocytes and spleens from leukemic patients. The enzyme and its template are localized in a particle having a density between 1.16 and 1.19 g/ml. These observations describe characteristic features of RNA tumor viruses.


Growth parameters of the hairless mouse epidermis after bleomycin treatment: possible interaction with epidermal chalones. Hairless mice were given two injections of 2 mg bleomycin i.p. on two successive days. By a stathmokinetic method, micro-flow fluorometry and autoradiography, many kinetic parameters were measured. Bleomycin affected cell proliferation in epidermis by depressing the number of cells in and the passage of cells through the different cell cycle phases, i.e. S, G2 and M in a biphasic manner. The time between the two minima corresponded to the mean generation time of the basal cells. Skin extracts produced from animals at different time intervals after the bleomycin injection show that bleomycin interferred only slightly with the G1-chalone, but had a more pronounced effect on the content of G2 of the skin, increasing and reducing the chalone effect in a biphasic manner. The study gives some support to the theory that the particularly good effect of bleomycin in squamous cell epithelium may be due to an interference of bleomycin with keratinization and chalone production. The protein synthesis showed only a minimal depression where the kinetic parameters were heavily depressed.


Structure and action of bleomycin.

Structure and action of bleomycin. The structures of bleomycins and of other bleomycin-phleomycin group of antibiotics were described. The activity of bleomycins and their derivatives in causing strand scission of SV40 viral DNA suggests that the beta-aminoalanine amide moiety and the carbamoyl group are involved in this reaction. More than one guanido group in the terminal amine of bleomycin-phleomycin group antibiotics caused irreversible renal toxicity in dogs. Pulmonary toxicity varied depending on the terminal amines. A bleomycin-inactivating enzyme which distributes widely in animal cells was shown to be a new aminopeptidase B which hydrolyzes beta-aminoalanine amide group. At least one of the reasons for activity against squamous cell carcinoma was shown to be due to the lower content of this enzyme. The inhibitor of this enzyme was synergistic to bleomycin in inhibiting growth of cells, thus suggesting the intracellular action of this enzyme. Selected for further study from the bleomycins containing various terminal amines, bleomycin 5033 which showed the same activity against squamous cell carcinoma in mouse skin as the bleomycin used at present and lower toxicity than the latter, and bleomycin A5196 which showed stronger activity and stronger toxicity but lower lung toxicity than the latter.


Importance of experimental data for the improvement of the therapeutical effect of bleomycin. The intimate mechanism of action of bleomycin has been extensively studied at molecular and cellular level. From the analysis of dose-response curves, different hypotheses have been formulated, and the importance and the extent of DNA repairability has been emphasized in correlation with the different phases of the cell cycle. Sensitivity to bleomycin depends on the fixation of DNA damage which occurs when damaged sites become involved in replication before they can be repaired.


Bleomycin effects on malignant tumors of the male genitalia. Bleomycin was applied alone or in combination with actinomycin D, vinblastine, radiation and/or surgery in cancer of the penis (6) and testis (6). In early stages of penile cancer (T2 N0 M0) a total response was noted. The failure in advanced stages is discussed. In terato-, embryonal- and choriocarcinomas of the testis (T2-4 N0-2 M0-1) remarkable responses of short and long duration were observed in all patients. The reasons for different biological action are discussed.


Effect of bleomycin on DNA, RNA, protein, chromatin and on cell transformation by oncogenic RNA viruses. Bleomycin (BLM) exclusively affects thymidine-containing compounds such as DNA and polydeoxyribonucleotides by releasing free thymine and leaving aldehyde functions. Molecular morphology and base sequence of the DNA strongly influence BLM activity. High BLM concentrations, besides modifying DNA into oligothyminic or athyminic nucleic acids, cause strand scissions. Enzymatic DNA and RNA synthesis is strongly influenced by BLM. The inhibition in DNA-dependent DNA polymerase and DNA-dependent RNA polymerase assays is of the non-competitive type. Protein biosynthesis in in vitro systems is not affected by BLM even at high concentrations. BLM turns out to be a strong inhibitor of DNase I and of DNase II; the inhibition is of the competitive type. The enzymatic activities of nucleases using RNA as substrate (RNase A, RNase B, Rnase T1, venom phosphodiesterase I and spleen phosphodiesterase II) are not influenced by this antibiotic. The antibiotic reduces cell proliferation (L5178y mouse lymphoma cells) in vitro in low concentrations by cytostasis and at higher concentrations by cytotoxicity. In BLM-treated L5178y cells, DNA synthesis is strongly reduced, while RNA and protein synthesis are not affected. In vivo, using growing quail oviducts, cell proliferation and cytodifferentiation are markedly inhibited after BLM treatment. This is attributed to the observed inhibition of DNA synthesis. RNA and protein synthesis as well as gene expression are not influenced by BLM under the conditions used. The selective inhibition of DNA synthesis in vivo may be caused by the following mechanisms: (1) competition of BLM with RNA; (2) blocking of the accessibility of DNA in chromatin to BLM, and (3) dependence from the repair processes. BLM inhibits growth of sarcomas, induced by oncogenic RNA viruses in vivo; well-developed tumours show regression after BLM treatment. Transformation of chick embryo fibroblasts by oncogenic RNA viruses in vitro and growth of these viruses is blocked by BLM; the most sensitive period for BLM inhibition is the time during the first period (integration of viral genome into cellular genome?) after infection.


Actions of bleomycin on DNA ligase and polymerases. Bleomycin inhibited the ligase, which was partially purified from rat ascites hepatoma, AH-130, even at a concentration as low as 0.01-1 mug/ml. The DNA degraded by bleomycin was not repaired by ligase. Therefore, it was suggested that bleomycin at higher concentration produced strand scission of DNA, which could not be repaired by the ligase, and at lower concentration inhibited the ligase reaction presumably by binding to DNA strand or to ligase. Also, the specificity of inhibition by bleomycin on the DNA polymerase of oncogenic RNA virus was tested, comparing with the four kinds of DNA polymerases extracted from the spleen of mice infected with Friend virus. Three kinds of DNA polymerases from spleen were not inhibited by bleomycin, but the fourth enzyme, which was induced in the spleen by virus infection, was inhibited by the antibiotic, when poly-d(AT) and poly-dG with dC were used as template.


Effect of bleomycin on cell survival and some implications for tumor therapy. Tumor is normally composed of cells growing and non-growing and, therefore, of cells having different responses to exogenous agents. Hence, it is not possible to treat a tumor with an unitary principle. As far as the tumor cell sterilization by bleomycin is concerned, the growing fraction of tumor should be managed by fractional schedule, while plateau phase cells by continuous infusion. Based on the cell sterilization kinetics, further improvement of chemotherapy by bleomycin must be sought. The investigation will duly involve the studies on the nature of resistance induction and PLD repair.


Response of mammalian cells to bleomycin-induced potentially lethal and sublethal damage. Both dividing and nondividing cells were able to recover from bleomycin-induced potentially lethal damage, but not from sublethal damage. The recovery from bleomycin-induced potentially lethal damage by nondividing cells was twice as great as that exhibited by dividing cells; and, potentially lethal damage recovery was sufficiently high for cells in both growth states to conceal the true response to sublethal damage. Through the use of integral dose fractions of bleomycin, the recovery from potentially lethal damage can be reduced significantly. These data suggest that closely spaced treatments with fractionated doses of bleomycin may be a more effective means of eradicating tumors containing large nondividing cell populations in vivo.


Sensorimotor organization in reach and prehension: a development model. A model is presented to depict the organization of the reflex, visual, voluntary motor, and perceptual-cognitive systems in the development of reach and prehension during the first year of life. Five components of hand skills are identified based upon a review of infant scales and observational studies, and the sequential occurrence of behaviors leading to the development of these components is graphically presented. These sequences are juxtaposed to enable the reader to view the temporal relationship of the systems at various stages in the development of reach and prehension in the infant. Items in the models are operationally defined. An explanation is included to clarify the nature and the sequence of events involved in the culmination of mature hand skills. The model should be useful in the identification, analysis, and remediation of deficits in reach and prehension often noted in children with developmental disabilities.


Mental development of phenylketonuric children on or off diet after the age of six. Eleven children with phenylketonuria (PKU) taken off diet after the age of six showed significant decreases in rate of mental development compared with 26 control children children of comparable IQ and with 17 PKU children of comparable IQ who remained on the diet. Changes in rate of mental development were significantly and inversely correlated with plasma phenylalanine levels in treated PKU.


[Competitive inhibition of the fixation of various cytotropic antibodies to mast cells of the peritoneal serous membrane in the guinea pig]. The degranulating capacity of guinea pig peritoneal serous membrane mast-cells due to IgG1 and reaginic homocytotropic antibodies and heterocytotropic rabbit IgG antibody was studied, as was the interference which nonspecific immunoglobulins may produce in the fixation of antibodies. Although the capacity to degranulate mast-cells proved identical for all the antibodies studied, reagin appears to possess a greater fixation capacity for a possible cellular membrane site. The greater resistence of reagin to elimination by washing could account for this.


[Study of serologic cross reactions between the antigens of Paracoccidioides brasiliensis and Histoplasma capsulatum]. A comparative study of serologic cross-reactions between histoplasmosis and paracoccidioidomycosis was carried out using complement fixation (CF) and agar gel immunodiffusion (ID) tests. Sera from 77 patients with paracoccidioidomycosis, 38 with histoplasmosis and 50 healthy individuals were studied. The Pan American Health Organization microtiter CF test and the Hupper ID technique were employed. The antigens were prepared by the method of Ajello as modified by Negroni. Paracoccidioidin CF test was positive in all patients with paracoccidioidomycosis; 29 of these (37%) presented serologic cross-reaction with histoplasmin. Histoplasmin CF test was positive in all but one of the patients with histoplasmosis and only 4 of them (10%) showed cross-reaction with paracoccidioidin. Positive reactions with the ID test were registered in 74/77 cases of paracoccidiodomycosis, with only one case of cross-reaction (1,3%). The result for the ID test was positive in all but one of the cases of histoplasmosis, with only one cross-reaction with paracoccidiodin (2,7%). The ID test seemed to be much more specific although it could be less sensitive. It was also easier to perform and less expensive than the CF test. Nevertheless the authors urge that the CF test also be carried out whenever posible, since the information obtained is likely to be of more value in follow-up and control of patients.


Confirmation of the existence of human serum leukaemia-associated antigens (LAA). We have obtained antisera from rabbits which react with serum from several leukaemia patients after absorption with normal human serum. The specificities of these rabbit antisera have been shown to be closely related to those of the original anti-LAA antisera of Viza et al (1970) and Harris et al (1971). Thus, the existence of leukaemia associated (but probably not leukaemia specific) serum antigen in man has been confirmed. One of the animals was immunized with normal amniotic fluid obtained at 15-16 weeks of gestation. Thus, LAA seems to be a normal constituent of amniotic fluid. This suggests that LAA is another onco-fetal component.


The immunosuppressive role of alpha-fetoprotein during pregnancy. Cell-mediated and humoral immune responses were assessed in mice at mid-term (day 10) in pregnancy. A significant but selective suppression of the primary in vivo antibody (plaque-forming cell) response to SRBC was observed, with the most pronounced effect being on the gammaA response. Similar results were obtained for secondary in vitro antibody synthesis by antigen-primed spleen cells from pregnant mice, demonstrating the intrinsic nature of the inhibition. Pregnant mouse serum (PMS) was shown to suppress primary in vitro antibody synthesis, and the inhibitory effect was abrogated by the selective removal of alpha-fetoprotein (AFP) using affinity chromatography. Normal mouse serum became similarly suppressive in vitro when purified AFP of fetal origin was added to it in concentrations approximating that found in PMS. Spleen cells from pregnant mice showed a suppressed mitogenic response to phytohemagglutinin, a lowered response to concanavalin. A, and a normal response to lipopolysaccharide. In contrast, the allogeneic response of these animals as measured in the one-way mixed lymphocyte culture was enhanced. PMS suppressed both allogeneic and mitogen-induced lymphocyte transformation by spleen cells from nonpregnant mice, and the effect was eliminated by the selective removal of AFP. These findings indicate an important functional role for AFP in normal embryological development.


The first example of an isoallotype of human IgG located on the Fd fragment. An isoallotype of IgG is described that is an isotype on heavy chains of IgG3 and IgG4 proteins and an allotype for IgG1 proteins, in which it is associated with Glm(f). The isoallotypic determinant is located on Fd. Binding of light chains (independent of their type) is required for its antigenic expression, since isolated heavy chains are not reactive in inhibition tests. It is suggested that the coding for the amino acid sequence related to this isoallotype was present on the ancestral gene before the duplication of subclasses. The presence in many non-human primate species proves that it is strongly conserved in evolution.


[Diagnostic methods in drug allergies].

[Diagnostic methods in drug allergies]. Immunological exploration of a patient following an allergic complication is based on tests in vitro, which are free of danger. At the present time it remains very difficult, on the one hand on the basis of the antigenic radicle responsible, secondly because of the characteristics of the immune response which it induces. The aim of exploration is the objective demonstration, using several techniques, of an immune response directed specifically against the medication. The pathological significance of this response must be analysed in each case.


The concentration of doxycycline in human prostate gland and its role in the treatment of prostatitis. The concentrations of doxycycline were measured in 36 enucleated, enlarged prostate glands. After administration of 200 mg of doxycycline intravenously significant tissue concentrations (mean value 1.79 mug/g) were rapidly obtained. The mean concentration of the drug 14 hours after a repeated oral dose of 100 mg was 1.56 mug/g. The ratios of tissue and serum concentrations were 0.610 and 0.568, respectively. As there are only a few antibiotics which penetrate into the prostate, the importance of these data for the treatment of bacterial or mycoplasma prostatitis is obvious.


Experimental production of rheumatoid factor-like antibodies and antibodies against the cathepsin D site of IgG following the injection of autologous Fab2. In a very high proportion of rabbits, repeated intra- or extra-articular injections of autolous Fab2 produced by homologous cathepsin D induce the formation of Rf-like antibodies reacting with both homologous and human IgG. Moreover, intra-articular injections of this kind cause a significant rise in the titre of thm of all the animals so far tested. Rf-like antibodies against human IgG appear earlier and have higher serum titres than those reacting with homologous IgG. The reason for this latter observation seems to be the blocking of the anti-rabbit IgG antibodies by the animal's own IgG. The anti-rabbit IgG antibodies can be absorbed only on aggregated rabbit IgG. The anti-human IgG antibodies cross-react to some extent with rabbit IgG. The results of inhibition studies suggest that the formation of anti-Fab2 homoreactants is directly stimulated by the injected Fab2, whereas the Rf-like antibodies owe their appearance to immune complexes formed in vivo by the injected Fab2 and the naturally occuring anti-Fab2 homoreactants. In respect of immunoglobulin class, the two kinds of Rf-like antibody are possibly of both IgM and IgG type.


[The importance of clinical observations for medical research]. Medical progress owes a great deal to the fundamental medical sciences and to the application of chemistry, physics and mathematics to medical problems. However, clinical observations and investigations are still of decisive importance in any field of medicine. By a feed-back mechanism they may even stimulate and fertilize fundamental medical sciences. Thus, our knowledge of the blood coagulation mechanism has been considerably enlarged by clinical analysis of hereditary bleeding disorders. - Chemotherapy of neoplastic diseases started from clinical observations during World War I (production of leucopenia by sulfur mustard gas). - Surgical procedures and their consequences have contributed greatly to our knowledge of thyroid function, of the segmental anatomy of the lung, and of the conduction system of the heart. - Observations of side effects of drugs have often enlarged or completely changed their primary clinical indications: from antibacterial sulfonamides, anti-diabetic, antihypertensive and powerful diuretic drugs have been developed, and from histaminics the modern neuroleptics and antidepressants. - Fundamental immunology has been enormously activated by clinical transplantation of kidney and bone marrow. Selective immunological defects in men, real experiments of nature, contributed much to our knowledge of the various types of allergic response. The quality of clinical investigations, particularly of controlled clinical trials, has been considerably improved during the last two decades. Although it is an applied science the reliability of its results is to-day comparable with that of "pure" natural sciences. However, medicine is more than a natural science: examples of outstanding scientists who at the same time were great and human physicians are presented.


Salivary gland hyperglycemic factor: an extrapancreatic source of glucagon-like material. Extracts of homogenates of rat, mouse, rabbit, and human submaxillary salivary glands contain a significant quantity of a material with glucagon-like immunoreactivity. Fractionation of this material on columns of Sephadex G-100 reveals a single peak immediately following a gamma globulin marker but in advance of a rat growth hormone marker, crystalline amylase, and isotopically labeled porcine insulin and glucagon. This material, which is urea stable, shows identical immunoassay dilution curves when measured with the highly specific K-30 glucagon antiserum. Study of paired glands in vitro shows that low concentrations of glucose stimulate and high concentrations of glucose suppress release of this material. Arginine promotes brisk release in vitro. Somatostatin does not influence arginine-stimulated secretion and insignificantly suppresses basal release in vitro. These findings lend support to previous speculations that the salivary glands may possess endocrine as well as exocrine functions. Salivary gland glucagon may also be the source of circulating glucagon recently reported in pancreatectomized and eviscerated rats.


Oncornavirus: isolation from a squirrel monkey (Saimiri sciureus) lung culture. An oncornavirus isolated from a squirrel monkey (Saimiri sciureus) lung culture has a density of 1.16 to 1.17 grams per milliliter, contains 70S RNA, and has an RNA-directed DNA polymerase that prefers Mg2+ over Mn2+ in an assay in which polyribocytidylate - oligodeoxyguanylate (12-18) is used as a synthetic template. Morphologically, the virus resembles Mason-Pfizer monkey virus but is antigenically distinct from this virus. The virus grows in cells of human, chimpanzee, rhesus monkey, canine, and mink origin, but not cells of squirrel monkey origin. On the basis of its properties, the newly isolated virus can be classified as a retravirus.


Reverse banding on chromosomes produced by a guanosine-cytosine specific DNA binding antibiotic: olivomycin. Characteristic reverse fluorescent banding patterns (R bands) on human, bovine, and mouse metaphase chromosomes are produced by treating chromosome preparations directly with olivomycin. With the DNA in solution, the repeating polymer poly[d(G-C)] - poly[d((G-C)] (where G is guanine and C is cytosine) enhanced the fluorescence of olivomycin, while the antibotic fluorescence was not affected by the alternating polynucleotide poly]d(A-T)] - poly[d(A-T)] (where A is adenine and T is thymine). Calf thymus DNA, with an intermediate G-C content of about 40 percent, showed a smaller fluorescence enhancement in the presence of olivomycin as was observed for the synthetic polynucleotide poly[d(G-C)] - poly [d(G-C)]. The closely related antibiotic chromomycin A3 showed the same results as were obtained with olivomycin either in the solution interaction with specific DNA's or with the netaphase chromosome preparations. The production of R bands by these G-C-specific DNA binding antibiotics lends credence to the suggestion that the arrangement of the nucleotide sequences along the chromosome is a primary determinant for the appearance of fluorescent bands.


Markers for ovarian cancer: regan isoenzyme and other glycoproteins. Since embryonic genes are not generally active in normal adult subjects and because certain of these genes are activated in cancer leading to ectopic synthesis, it is the difference between the ectopic level and the normal adult concentrations of embryonic gene products which we seek in developing "markers" for ovarian cancer. The carcinoplacental alkaline phosphatases corresponding to the term gestational phenotypes correlate positively with ovarian cancer as does hCG. Other fetal and placental glycoproteins whose presence is noted in ovarian cancer include CEA, alpha-FP, and Björklund's antigen. Antigens of mucinous cystadenocarcinoma have not yet been examined for their possible fetal or placental origins. The degree of concordance of expression of Regan isoenzyme and hCG is variable. Profiles of glycoproteins would appear to offer an opportunity to inquire more deeply into the nature of ovarian cancer and from this inquiry, one can expect to develop a system of markers which can be of clinical use.


Establishment of a continuously growing cell line from primary carcinoma of the liver. A continuously growing cell line has been established from autopsy material taken from a primary liver carcinoma, a cancer which has a high incidence in Southern Africa. The cell line was initiated from multifocal areas of outgrowth in the primary culture, and adaptation to in vitro conditions was completed after 18 months. The cells resemble hepatocytes in culture, have a doubling time of 35-40 hours and a plating efficiency of 40-50%. No virus particles have been found in the cells by ultrastructural examination. The isozyme pattern and the karyology of the cells are human. The chromosome pattern is heteroploid (mean number 56) and there are marker chromosomes.


An optical switch for improved photography through the operating microscope. An optical switch is described which allows utilization of both a 35 mm still camera and a video camera on the operating microscope. This allows more versatile photodocumentation without sacrificing the other port on the beamsplitter which would normally be utilized by the assistant.


A simple fluorescence staining technique for the differentiation of human tissue transplanted into nude mice. Human and mouse nuclei can be distinguished by differences in the constitutive heterochromatin when stained with quinacrine dihydrochloride. With the staining method described, mouse heterochromatin during interphase appears as brilliant fluorescent chromocenters. By replacing the commonly used aqueous buffer mounting medium with a xylene-diluted synthetic resin, the haziness of the nuclear fluorescence is eliminated thus allowing identification of the heterochromatin pattern in histological preparations. A requirement for the definite identification of cells of human or murine origin in the nude mouse is the knowledge that the heterochromatin arrangements changes according to the stage of differentiation of the cell of the position of a particular nucleus within the cell cycle.


Binding of [3H] methyltrienolone (R 1881) in rat prostate and human benign prostatic hypertrophy (BPH). Methyltrienolone (R 1881 - 17beta-hydroxy-17alpha-methyl-estra-4, 9, 11-trien-3-one) binding to rat ventral prostate cytosol has a specificity typical of an androgen receptor. In human benign prostatic hypertrophy (BPH) tissue, the specificity of [3H] R 1881 binding is different from that measured in rat prostate: progesterone and R 5020 (17, 21-dimethyl-19-nor-4, 9-pregnadiene-3, 20-dione) being more potent while 19-nortestosterone is less potent competitor. Moreover, the synthetic progestin [3H] R 5020 binds to BPH tissue with a similar specificity. These data suggest the presence of progestin binding components or of an atypical androgen receptor in human BPH cytosol.


Indian childhood cirrhosis: genealogic data, alpha-foetoprotein, hepatitis antigen and circulating immune complexes. Indian childhood cirrhosis is a significant cause of morbidity and mortality in young children in India. One hundred patients with ICC, 66 boys and 34 girls, were studied. Pedigree analysis yielded a segregation ratio of 0-2196, suggestive of an autosomal recessive inheritance. Serum alpha1-antitrypsin level was normal. Serum alpha-foetoprotein (AFP) concentration was increased in all the patients, parents and in some siblings. Serum immunoglobulins G, A, M, and D were elevated. Haemolytic complement and C3 were low. Electrophoretically altered complement components were detected in 36% of patients. There was an inverse relationship between C3 concentration and immunoconglutinin titre. Circulatingimmune complexes were detected in the sera of six out of ten patients who had significant proteinuria. Hepatitis B surface antigen (HBsAg) was present in the serum, ascitic fluid, saliva, urine and faeces of ICC patients more frequently than in controls. HBsAb was detected less often. Lymphocyte response to HBsAg was impaired. The first-degree relatives had a higher incidence of HBsAg and HBsAb than healthy controls. It is suggested that ICC occurs in infants with an inherited hepatocyte vulnerability and that one of the precipitating causes of liver cell necrosis is infection with hepatitis virus(es). The consequent immunologic epiphenomena contribute to progressive hepatic damage ending in death.


T cell-specific human alloantisera-detecting antigens segregating outside the major histocompatibility complex. Two sera demonstrated non-HLA lymphocytotoxicity on the basis of reactivity with the cells of siblings genotypically identical to the serum donors for the major histocompatibility complex. These two sera, Bl and Caf, once contaminating HLA antibodies were removed by absorption with pooled platelets, demonstrated allogeneic lymphocytotoxicity that was restricted to T lymphocytes. Reactivity of the absorbed sera segregated independently of the major histocompatibility complex in 3 of 12 families tested. Unlike both cold lymphotoxins and HLA antibodies, the absorbed sera showed little temperature sensitivity against allogeneic cells, although reactivity of the Bl serum to autologous cells and to cells of the donor's HLA identical sibling did show a decrease with increasing temperature and restriction of activity to the 19S-containing fraction. Granulocytes were unreactive with the absorbed sera. Such sera may provide probes of minor transplantation antigens or markers, or both, of lymphoid subpopulations.


Evaluation of cellular immunologic responsiveness in the clinical management of patients with prostatic cancer. I. Thymic-dependent lymphocytic blastogenesis. Thymic-dependent lymphocytic blastogenesis of peripheral blood lymphocytes of 59 patients with varying stages of prostatic cancer to the non-specific plant mitogen, phytohaemagglutinin (PHA) and the correlation of their responsiveness with the clinical stage of malignancy and level of alpha2-globulin have been evaluated. Patients within each of the four stages of malignancy possessed statistically significant extrinsic (noted in 40 (68%) of 59 patients) and intrinsic (noted in 21 (47%) of 45 patients) aberrations of their lymphocytic responsiveness to PHA compared with the responsiveness of a control population of non-cancer patients. The observed aberrations were, however, not significantly different between each stage nor did they correlate with the stage of disease. Similarly, levels of alpha2-globulin, while significantly elevated within each stage, as compared with the levels in the control population, no significant differences or correlation with the stage of disease was observed. Of interest, perhaps pending further study, were observations of the increased frequency of the number of patients with a significant elevation of alpha2 with a progression of malignancy from localized to invasive and metastatic disease. A similar trend in the incidence of the association of aberrations of lymphocytic reactivity with elevated levels of alpha2 were also noted with a progression of disease. The present confirmatory observations of a recent study in this laboratory of diminished cellular responsiveness in patients with prostatic cancer may be of considerable relevance in directing the therapeutic management of the patient - lest the therapy selected be further debilitating providing reduced surveillance - metastization of tumour cells, and alteration of tumour-host homeostasis.


[Electroresection at the apex of the prostate gland: aid for orientation by faradic stimulation of the external sphincter muscle (author's transl)]. The localization of the external sphincter muscle can be very well demonstrated by its contractions, which can be produced by transurethral faradic electrostimulation. We present a simple and safe method of stimulating the sphincter muscle if wished during TUR or for diagnostic reasons. This exact orientation-aid, which is simultaneously a function test, is more precise than the known "sphincter signs".


[Reliability of cytologic examination in the diagnosis of laryngeal carcinoma (author's transl)]. Cytologic examination has been performed as systematic survey, with drawing by plug in direct laryngoscopy, on 96 patients with intrinsic and extrinsic laryngeal lesions and, as control, on 22 patients with oro- and rhinopharyngeal lesions. The reliability of cytologic diagnosis in relation to the type of the lesion (histologically controlled), to the site of the lesion and to the staining method is evaluated.


The H specificities of butanol extracts of human erythrocytes. A previous report of other researchers that aqueous phase fractions of erythrocyte stromata of secretors contained more ABH blood group activity than similar fractions of non-secretors was confirmed. The recovery of H activity in the butanol phase extracts of both secretors and non-secretors, which had not been seen previously, is reported here for the first time. Three H specificities were present when the aqueous and butanol phases of group O secretor stromata were examined in inhibition of agglutination studies with anti-H of group Oh and group A1 sera and three lectins. The anti-H (group Oh) sera were inhibited by both aqueous and butanol fractions. Anti-H serum (group A1) and the lectin Ulex europaeus were inhibited by the aqueous fraction only. The lectins Laburnum alpinum and Lotus tetragonolobus were not inhibited by either the aqueous or butanol phase extracts.


Sterilized hepatitis B (surface) antigen for production of specific antisera. The combined treatment with beta-propiolactone and ultraviolet rays described for the sterilization of HBsAg-positive sera does not interfere with the purification procedures for the production of HBs antigen. The immunogenicity of the HBs antigen is retained.


[Antigenic characteristics of oncovirus persisting in continuous Syrian hamster cell culture]. The antigenic properties of an oncornavirus persisting in a continuous Syrian hamster cell culture (X-100) were studied. By the mutual immunodiffusion test it was shown that virions and virion-producing cells contained a protein antigenically identical to the main structural protein of oncornaviruses type C of Syrian hamsters. The materials tested contained no "GS-1" antigens (species-specific antigenic determinants of the main structural protein of mammalian oncornaviruses type C) of murine, feline, simian viruses or group-specific antigens of Bitner and Mason-Pfiser viruses.


[Preparative isolation of alpha 2-macroglobulin, transferrin, albumin and study of their nonspecific gamma-inhibitory activity]. Profiles of distribution of non-specific gamma-inhibitors of influenza A2/Victoria/35/72 in donkey and horse sera were established by gel chromatography in Sephadex G-200. High and low molecular inhibitors were found in 19S and 4S serum fractions. Highly purified preparations of a2-macroglobulin, transferrine and albumin were isolated by a combination of methods of salt precipitation, gel chromatography on Sephadex G-100, G-200 and ion exchange on DEAE-Sephadex A-50. Heating sera resulted in a considerable increase of the antiviral activity of a2-macroglobulin and transferrine and a reduction of albumin activity.


[Various biological properties of diploid cell strains derived from various tissues of human embryo]. Some properties of 4 diplod cell strains derived from the lung and skin-muscle tissue of human embryos were studied. Only one strain (L-63) was shown to have a stable karyotype throughout its life span and induced no tumor formation in the cheek pouch of hamsters. Species-specific and group antigens were revealed. In the cell strains of the lung and skin-muscle derivation differences in the distribution of lactate dehydrogenase isoenzymes were observed and found characteristic of the original tissue cells.


[Characteristics of encephalomyocarditis virus isolated from sick monkeys]. A cytopathic filterable agent designated EMC-70 was isolated in 1970 from Macaca rhesus monkeys with encephalomyocarditis. Its biologic and physico-chemical properties were studied. A number of primary and continuous cell cultures were found to be susceptible to the virus, and it was pathogenic for monkeys and small laboratory animals but did not multiply in chick embryos. The virus was stable to chlorofrom, thermolabile, stable to acids, and agglutinated group O erythrocytes of man, sheep, guinea pig. Electron microscopic examinations of infected cells revealed crystalline packings of subunits about 50 nm in diameter typical of picornaviruses. Serological studies demonstrated close antigenic relationship of the isolate with the virus belonging to encephalomyocarditis group.


[Various properties of virulent and moderate Erwinia carotovora phages]. A bacterial culture of E. carotovora 8638 lysed by virulent phage 62 after treatment with UV rays produces a moderate phage. The virulent and moderate viruses, antigenically unrealted, have a similar prolonged latent period (100 min) and the period of lysis (60 min). They also differ in the sizes of the capsids and the length of processes which are 750+/-30 and 2000+/-50A in the virulent phage and 600+/-30 and 1500+/-50A, in the moderate phage, respectively. The sedimentation constants are 602 and 340 S, respectively. In the virulent virus, the volume of the cavity of the protein capsid contianing 8 major and 5 minor peptides is twice as large as that in the moderate one (with 8 peptides); the molecular weight of their DNA shows approximately similar ratios: 52.53+/-17X10(6) and 29.79+/-0.88X10(6) daltons. Evidently, the moderate phage because of the reduced size of the genome contains less message for coding for structural proteins and nucleoprotein as a whole.


[Comparative antigenic characteristics of 2 Marek's disease virus variants and of herpesvirus of turkeys isolated in the USSR]. The antigens induced by pathogenic (55) and nonpathogenic (83) variants of Marek's disease virus, the Kekava strain (MDV-Kekava) and herpesvirus of turkeys, the M-24-72 strain (HVT-M24-72) were tested in the agar gel precipitation test. The antigens were prepared from feather follicles (FF) of the infected chickens, from the infected cultures of chick embryo fibroblasts (CEF) and from the culture fluid (CF) of these culttures. In the FF and CEF cultures infected with MDV-Kekava-83, the synthesis of the antigens was 6-10 times as low as in the FF and CEF cultures infected with MDV-Kekava-55. Both variants of MDV-Kekava retained their capacity for antigen synthesis in the CF up to 84 passages. Three antigens or three groups of antigens were found in the preparations examined: (1) strong common antigens in the CF and FF preparations; (2) weak common antigens in cell preparations and CF; (3) strong antigen(s) in cell preparations not identical to the CF and FF antigens. In CEF culture preparations infected with MDV-M-24-72, using sera from chickens with Marek's disease an antigen was detected partially identical to the weak antigen of cell preparations and cross-reacting with strong antigens of the FF, CF and cell antigens of MDV. No antigen was found in concentrates of CF from the CEF cultures infected with MDV-M-24-72 with the sera used. It is suggested that the CF and FF MDV antigens are virus antigens, while antigens of the cell preparations of this strain are autoantigens or embryonal antigens. Data are presented on relationship between the MDV-Kekava antigens with the MDV-HPRS-16 antigen.


[Cellular immunity responses in studies of the specificity of leukocyte hepatitis virus]. Hypersensitivity of a delayed type to leukocyte hepatitis virus (LHV) was established in patients with virus hepatitis. The results of the skin tests with the LHV antigen were positive in 15 out of 20 patients with active chronic hepatitis, in 17 out of 20 convalescents and in 3 out of 20 patients in the acute stage of virus hepatitis, and negative in 4 normal subjects and in 12 out of 14 patients with chronic hepatitis in the stage of stabilization. The results of the skin tests with the antigen of donor leukocytes were negative in 130 patients with hepatitis and normal subjects. In the leukocyte migration inhibition test, 1/4 of the patients with chronic hepatitis were found to be sensitized to the LHV antigen, in the absence of sensitization by this test to the HB antigen and the antigen in the plasma of hepatitis patient.


[Changes in cell mitosis under the influence of antigenically different strains of respiratory syncytial virus]. A peculiar effect on mitosis of RH cells of the reference Long strain and strain No. 14 of respiratory syncytial virus isolated during a rise of the incidence of acute respiratory diseases in Kharkov was observed. The Long strain affected mostly the qualitative and strain No. 14 the quantitative aspects of cell mitosis. The character of pathological mitoses under the influence of the Long strain indicated more considerable impairment of the mitotic apparatus of the cells than that caused by the strains No. 14. It is suggested that the peculiarities of the effect of the two respiratory syncytial virus strains on cell mitosis are associated with the structural and antigenic differences in the strains circulating in various geographic zones.


The effect of bleomycin on mitochondrial DNA. Covalently closed circular mitochondrial DNA (mt-DNA) from mouse fibroblasts (L-cells) forms a separate band from nicked circular or linear DNA molecules in ethidium bromide-cesium chloride byoyant density gradient analysis. The conversion of covalently closed circular mt-DNA to nicked circular or linear DNA may serve as a sensitive method of detecting DNA strand breaks induced by bleomycin (BLM). At a very high concentration BLM induces the conversion of closed circular mt-DNA to nicked circular or linear DNA, however even after 24 h of exposure to BLM at a high concentration a considerable amount of closed circular mt-DNA molecules remains intact.


[Intraarterial bleomycin therapy on squamous cell carcinomas of the oral cavity. Clinico-pathological investigations (author's transl)]. The mode of action of bleomycin after intraarterial perfusion in 53 patients with advanced squamous cell carcinoma of the oral cavity is reported on the basis of clinical results and pathologico-anatomic findings. In 29 out of 32 cases not previously treated a clinically complete remission could be achieved by primary chemotherapy. In 8 cases these findings were histomorphologically confirmed and 7 patients have shown a persistant remission for 9 to 61 months. The chances of success are much less favorable in the treatment of recurrences. No clinical advantages were observed when combining bleomycin with other chemotherapeutics.


[Nucleus tuberis lateralis in the human brain. A pigmentarchitectonic study]. In the present article the shape of the nucleus tuberis lateralis and a hitherto unknown cell group in the human hypothalamus is described. Neurolipofuscin is selectively stained with aldehyde-fuchsin. This method enables the exact determination of the three dimensional shape of a griseum by using slices more than 500 mum thick. Differences in the distribution and amount of pigment within different types of nerve cells provide a basis for cellular demarcation and subdivision (pigmentarchitectonics). The intensely stained nucleus tuberis lateralis can be seen as consisting of a main lateral and a medial group. There are elevan cell groups which constantly occur. The individual range of their shapes causes a great variation in the shape of the lateral tuberal nucleus. It also has portions in the medial field of the hypothalamus: a pars suprafornicalis and 'mediale Streuzellen' ('medial scatter-cells'). Rostrally it extends almost to the chiasma with scattered irregularly occurring groups of cells. The amount of intracellular lipofuscin depends on the respective cell group and the age of the brain. By the age of 12 years the nucleus is already more intensely pigmented than the surrounding grisea. There is an indication of a tendency towards an aggregation of the cell groups with age. Connections between pigmentation and the phylogeny are discussed. Ventrally the nucleus tuberis lateralis  is bounded by a hitherto undescribed heteromorphous cell group. It is clearly delimited and can be divided into a longish pars lateralis and a spherical pars medialis. Both parts consist of small as well as medium-sized cells which accumulate diffusely distributed granula of lipofuscin in adult brains. The name 'nucleus paratuberalis' is suggested for this griseum.


Pharmacokinetic study with synthetic salmon calcitonin (Sandoz). 18 patients randomly divided into 3 groups of six each received 35 mug (140 M.R.C. Units) of synthetic salmon calcitonin intravenously, intramuscularly or subcutaneously. Plasma and urin concentrations were determined using the radioimmunoassay method. There was a rapid distribution phase of ca. 12 minutes after intravenous injection then an elimination half-life of 1.1 hours. The volume of distribution was 11 litres. The invasion half-life after intramuscular and subcutaneous administration was 12 and 11 minutes respectively and the elimination half-lives 1 and 1.5 hours, respectively. The bioavailability of the intramuscular and subcutaneous forms was found to be 66 and 71% respectively when areas under their plasma concentration/time curves were compared with the intravenous area.


Does salmon calcitonin influence the motility of the human gastrointestinal tract? An electromanometric and endoscopic study. The effect of salmon calcitonin--as a relatively high dose of 25 mug by single iv. injection--upon the basal and metoclopramide-stimulated motility phenomena of the stomach and proximal duodenum as well as upon the unstimulated colon and rectosigmoid was evaluated in 12 subjects without gastrointestinal disease by quantitative electromanometric measurements performed for one hour with the aid of fluid-filled micro-balloon-catheter-systems. Additionally, in 9 subjects the reaction of the cardia, stomach, pylorus and proximal duodenum upon the same dose of the hormone was observed endoscopically for 30 minutes. Under the conditions of our study there was no evidence to be obtained for any measurable or visible effect of salmon calcitonin upon the tonus and motility of the smooth muscle organs of the human gastrointestinal tract.


Inhibition of pancreatic secretion of enzymes by calcitonin. Calcitonin in pharmacological dosage inhibits the secretion of enzymes in the human pancreas without influencing the production of fluid and bicarbonate. The degree of inhibition is the same in a juice stimulated by secretin alone and by secretin plus pancreozymin-cholecystokinin or caerulein. Thus the endogenous and the exogenously applied pancreozymin is inhibited in its action. The secretion of enzymes, which is induced cholinergically by carbamylcholine or insulin hypoglycemia, is not altered by calcitonin. The inhibitory action of CT is not counteracted by infusion of calcium thus showing that local depletion of calcium is not the mechanism of inhibition. Possible mechanisms are the direct interference with pancreozymin at the acinar cell and the inhibition of hormone-release from the pancreozyminproducing cells.


Inhibition of gastric acid and gastrin secretion by somatostatin in man. Somatostatin inhibited gastric acid secretion significantly in three ZE patients, while serum gastrin was lowered only in two. A direct inhibition of pentagastrin-stimulated acid secretion by somatostatin was demonstrated in three healthy volunteers and four patients with duodenal ulcer disease. Somatostatin reduced betazole-stimulated gastric acid secretion to a lesser degree. The immunohistological demonstration of somatostatin-producing cells apposite to antral G-cells suggest a physiological role of somatostatin in the regulation of gastrin secretion in man.


Inhibition by somatostatin of pancreatic juice and enzyme secretion and gallbladder contraction induced by secretin, cholecystokinin-pancreozymin and carbachol administration. The effect of somatostatin on secretin-, CCK-pancreozymin-and-carbachol-stimulated pancreatic secretion was studied in eight healthy volunteers. In all instances there was a significant reduction in duodenal secretory volume after secretin without any change in bicarbonate concentration. CCK-parcreozymin and carbachol stimulated volume and enzyme secretion were significantly inhibited and gallbladder contraction was abolished by somatostatin. In three volunteers a reversible hypercoagulation during infusion of 200 mug/h cyclic somatostatin has been observed. Somatostatin inhibited the CCK-pancreozymin stimulated pancreatic enzyme secretion of anaesthesized rats.


Effects of synthetic salmon calcitonin on gastric secretion and ulcer formation in conscious cats and rats. In conscious and unrestrained cats the gastric secretion and ulcer formation induced by i.v. infusion of pentagastrin can be inhibited by synthetic salmon calcitonin given simultaneously. The volume and various constituents of gastric juice are proportionally diminished. Pepsin alone is definitely more inhibited, which may be of significance in respect to the mode of action of calcitonin. These effects are dose dependent in the range of 0.01 mug - 1.0 mug/kg/h salmon calcitonin, corresponding to 0.05 - 5.0 MRC units. Based on the finding that such minute doses have effects, it may be speculated that calcitonin has a regulatory function in gastric secretion of cats. In Shay-rats a dose dependent inhibitory effect of salmon calcitonin on ulcer formation and gastric secretion is demonstrated. Besides the volume, the acid concentration of gastric juice is reduced, which may explain the high efficacy of salmon calcitonin to prevent ulcer formation in this species. Ulcerations induced by pylorus ligation, stress and phenylbutazone can be inhibited to a similar degree by calcitonin, suggesting interference with a basal mechanism common to all three types of ulcerogenesis.


Calcitonin-biochemistry and physiology.

Calcitonin-biochemistry and physiology. Calcitonin is still a mysterious protein hormone. Secreted by the C-cell in ultimobranchial body or thyroid glands, it seems to act as a hypocalcemic antagonist against parathyroid hormone by inhibiting osteolysis. This concept which was developed especially in small mammals like the rat, does not entirely fit the results gained from other species: in fish, CT levels change in connection with the reproductive cycle in the female, in amphibians, buffering capacity of the organism is bound to the ultimobranchial body, and in birds, sexual development in the male is accompanied by increasing calcitonin levels.


Long-term effects of calcitonin on gastric secretion in normals, peptic ulcer and high risk patients. Using a 12-hour infusion of salmon synthetic calcitonin (S-CT), distinct and sustained inhibition of gastric acid and pepsin secretion has been demonstrated in 4 normal subjects, 3 patients with peptic ulcer disease and 3 high risk patients. In 3 patients with Zollinger-Ellison syndrome, treated in the same way, elevated serum gastrin was reduced by about 50% and acid secretion by more than 90%. In healthy volunteers oral administration of human synthetic CT (H-CT) led to reduction in basal and pentagastrin-stimulated acid and pepsin secretion by about 50%, lasting for more than 2 hours after the instillation of CT. In 4 subjects receiving CT intravenously, slight nausea and headache were registered, while there were no side effects after the oral route. Serum calcium did not change after i.v. or oral administration of CT. Wheras therapeutical applications of CT, given by i.v. route, seem to be restricted to selected cases, i.e. acute gastric ulcerations with imminent or existent bleeding, the eventual benefit or orally administered CT in peptic ulcer disease should be evaluated in controlled long-term trials.


[Current state of radiotherapy of mouth neoplasms]. Beside surgery and chemotherapy, which has recently become important, radiotherapy is an efficacious means in the treatment of oral cavity cancer, both when applied alone and in combination. Clinical radiotherapy is based on the differences in sensitivity between normal tissue and tumor. The radiosensitivity of oral cavity cancers--above all squamous cell carcinoma--may even be considered to be mediocre: therefore a dose should be reached in order to destroy the malignant growth. In percutaneous radiotherapy orthovoltage is without any prospects; high-energy methods, above all telecobalt, have certain advantages, particularly the lower absorption in the bones. In "brachytherapy" we prefer the artificial nuclides 137Cs and 60Co instead of 226Ra for moulds and 198Au grains for interstitial application. At first the radiotherapist has to decide whether to advise radical or palliative treatment. Patients with secondary nodes in whom "radical surgery is inadvisable should have radical radiation therapy..." (Gibb and Todd) in a dose of nearly 5000 rads. Radiotherapy after an inradical neck dissection is burdened with a therapy before surgery can be considered in select cases. The treatment of oral cavity cancer should be practiced in close cooperation between the surgeon and the radiotherapist.


[Epidemiological and laboratory criteria in the assessment of meningococcus carrier state]. A study was made of the duration of meningococcus carrier state, immunological indices and group-specific properties of meningococci isolated from them. The periods of meningococcus discharge were studied in 738 persons. Three categories of the carrier state were revealed: a single discharge (67% of the carriers), of average duration (up to 4 weeks), and prolonged. Greater indices of group-specific antibodies (in the passive hemagglutination test) were revealed in the carriers with prolonged presence of meningococci in the nasopharynx. Dynamics of immunological indices and periods of the carrier state formation in the foci of infection permitted to characterize the prolonged carrier state as a latent form of meningococcus infection. A study of the group-specific properties in 1845 strains and comparison of the group-specific pattern of the circulating strains with the epidemic situation indicated that meningococci of group A were not only epidemic, but also more virulent.


[Vaccine from the cell fragments of Bordetella pertussis. I. Protective, sensitizing properties and morphological characteristics of the vaccine]. The authors present the results of studying the protective and sensitizing properties of a new preparation made of a ultrasonic disintegrate of pertussis microbes treated by ethyl ether. As shown by electron microscopy, the preparation consisted of the cell wall elements (the membrane), remnants of the cytoplasm and protectosome, i.e. it represented a vaccine consisting of cell fragments. In crude and sorbed condition it possessed marked protective properties (a test on mice). The content of protective units in the adsorbed preparation increased 1.5-3 times. The vaccine produced no sensitizing action, and its histamine-sensitizing activity was 3-5 times lower by protein and 5-10 times--by IOU than that of the whole-cell vaccine prepared form the same microbial suspension.


[The effect of homologous gamma-globulin on nonspecific factors of antiviral immunity in prophylactic and therapeutic application]. Administration of homologous gamma-globulin for prophylactic purpose promoted an increase in the interferon-synthesizing activity of leukocytes. The therapeutic use of this preparation in tick-borne encephalitis produced no stimulating effect on this index.


[Identification with the aid of various serological reactions of L forms of streptococci isolated from the live organism]. Growth inhibition, agglutination, precipitation, and passive hemagglutination tests were used for the identification of the L-forms of streptococci isolated from the organism of experimental rabbits both after the infection with the L-forms of streptococci and with the streptococci of group A. The tests were positive not only with the antiserum of homologous, but also of heterologous strains of the L-form of streptococcus, group A. The L-form cultures isolated from the experimental animals failed to differ from the laboratory strain of the L-forms of streptococcus, group A, by serological properties.


A cytological method for the simultaneous staining of nucleoproteids and some cathionic proteins. A cytochemical method is suggested for the simultaneous and differential staining of cellular nucleoproteids [ribonucleoproteids (RNP) and desoxyribonucleoproteids (DNP)], as well as for the simultaneous contrast staining of some basic (arginine- and lysin-containing) proteins. The staining technique is based on DNA-denaturation procedures and the application of mixtures of basic dye--methylene blue and acid dyes--eosin or fast green at low concentrations. The combination of methylene blue with eosin is used for the staining of ribonucleoproteids (RNP) whereas methylene blue-fast green for the simultaneous detection of ribonucleoproteids and desoxyribonucleoproteids (RNP and DNP), as well as for the differential staining of nuclear DNP (after cold hydrolysis with 5 N HCl). The acid dyes eosin and fast green stain in pink resp. in green some cathionic proteins in the lysosomal (specific) granules of the neutrophilic and eosinophilic leucocytes in the cytoplasm of erythrocytes, and after cold hydrolysis in the cytoplasm of lymphocytes. A fluorescent variant of the method with sulfaflavin is also suggested for the fluorochromation of cytoplasmic cathionic granules in the luecocytes. Acid mucopolysaccharide components in the granules of basophilic leucocytes, tissue mastocytes and thrombocyres are stained intensively pink-violet (gamma-metachromatic). The possibilities for the application of the method in the quantitative analysis of blood and exfoliated cells, as well as for purpose of haematology, immunology and exfoliative cytology are discussed.


Changes in the mitochondria in the initial part of the axon during regeneration. The initial part of the axon including the axon hellock, the initial unmyelimated segment and the beginning of myelinated axon was studied electron microscopically during regeneration, 1--30 days following a crush lesion of the rat hypoglossal nerve. Large mitochondria reaching 1.1 mum in diameter, with abundant cristae and dense granules in the matrix were observed between days 3--21. They formed clusters in the initial myelinated segment of the axon. End-to-end contacts and ribosomes around them were very often visible. The large mitochondria exhibited strong succinate dehydrogenase and NAD - H2 diaphorase activities. The relationship between the appearance of large and active mitochondria in the initial part of the axon and the elevated axonal transport during regeneration of the peripheral nerve is also discussed.


Orcein staining of hepatitis B antigen (HBAg) in conventional paraffin sections of liver biopsies. In an unselected group of 213 liver biopsies, paraffin sections were treated with an orcein staining method with the view to demonstrate HBAg. In 8 patients with positive HBAg serology, only 3 biopsies yielded positive orcein material in hepatocytes. A good correlation appears to exist between orcein positive material and the "ground glass" cytoplasm of liver cells.


The cellular pathology of microgyria. A Golgi analysis. A case of extensive classical four layered cerebral microgyria was examined in whole brain serial section using routine cell and fiber stains, and in Golgi impregnations. Golgi preparations demonstrate that microgyric cortex contains the spectrum of neurons characteristically found in normal cortex, and that they are segregated by class into horizontal laminar comparable to layers I--VI in the normal. In microgyric cortex neurons of the mid-cortical layers, variably layers II--V, are replaced by a tangential band of fibrous astrocytes. These findings confirm impressions from general cell and fiber stains that classical four layered microgyria is the result of a destructive process striking predominantly in mid-cortical regions. It must occur after migration is complete but before the development of secondary and tertiary gyri, that is, between the fifth and seventh fetal months. The cellular pathology as seen in Golgi impregnations establishes that neurons surviving above the scar are normally differentiated, aligned, and oriented. By exception, many surviving neurons at the border of the scar have attenuated dendritic arbors which are oriented tangentially. Very few axons or dendrites actually traverse the scar.


Demonstration of the heterogeneity of the mast cell population on the basis of the mucopolysaccharide content. From the aspect of its mucopolysaccharide content the mast cell population is not homogeneous. The pulmonary and heart muscle mast cells of the rat are alcian blue positive, the mast cells of the thyroid gland, lymph nodes, subcutaneous connective tissue, mesentery and peripheral nerve are safranin positive, whereas among the mast cells of the peritoneal cavity and the thymus there are both alcian blue and saffranin positive forms. The least acid mucopolysaccharides are in the mast cells of the peritoneal fluid, the mesentery and the lungs, whereas the most acid ones are in the mast cells of the lymph nodes, the subcutaneous connective tissue and the thyroid gland. There is a considerable difference between the two last mentioned organs. The mast cells of the subcutaneous connective tissue are end-product cells without amine or precursor turnover, whereas the mast cells of the thyroid gland incorporate and deliver amines, which may participate in the regulation of the host gland.


Lysosomal enzyme studies after endotoxin administration in normothermia and hypothermia. The LD50 dose of endotoxin results in a considerable increase in the plasma level of acidic phosphatase and beta-glucuronidase. The endotoxin decreases the quantity of gamma-globulin fraction of sera as an effect of neutral lysosomal proteases. In hypothermic rabbits the activity of lysosomal enzymes is increased only slightly after administration of endotoxin and the change in the gamma-globulin level is also more less than in the normothermic animals. The importance of our results in the pathogenesis of inflammation induced by endotoxin is discussed.


Prenatal genetic diagnosis.

Prenatal genetic diagnosis. More than 100,000 children are born in the U.S. each year with major congenital defects or genetic disorders, with or without mental retardation. Advances in prenatal diagnosis have now made it possible to detect an increasing number of these disorders in utero. Carrier detection is advised prior to pregnancy, rather than after the birth of a defective child, or during pregnancy itself. Patients have a right to know about their genetic risks and should have the freedom to exercise their options.