Crystallographic structural studies of a human Fc fragment. II. A complete model based on a Fourier map at 3.5 A resolution. The crystal structure analysis of a human Fc fragment was pursued to 3.5 A resolution and a complete model was built and refined into the isomorphous Fourier map. The CH2 and CH3 domains show the immunoglobulin fold, with CH3 being closely similar to CH1, but CH2 intermediate in structure between V and CH3. The carbohydrate is rigidly attached to CH2, covering the C face. CH3 dimerizes as CH1-CL, but CH2 has no contact to the second chain. Residues involved in the lateral CH3-CH3 and the longitudinal CH3-CH2 contact are conserved in Ig classes and sub-classes. In IgM and IgE the two C-terminal domains also show this characteristic distribution of contact residues.


[On the occurence of isopeptide bonds in the chorion of the rainbow trout (Salmo gairdneri Rich.) (author's transl)]. Chorions of unfertilized and fertilized eggs of the rainbow trout were isolated from tissue and yolk components and investigated for isopeptide bonds. The complete hydrolysis of the alpha-amide bonds was obtained by a system of four proteases. A chromatographic ion exchange system was used to separate Nepsilon-(beta-aspartyl)lysine as well as Nepsilon-(gamma-glutamyl)lysine from single amino acids. Chorions of unfertilized eggs contain neither Asp Lys nor Glu Lys bonds. Only chorions of fertilized eggs contain Glu Lys isopeptide. It is probably the high content of this isopeptide that is responsible for the greater mechanical stability of the fertilized teleostean egg chorion.


Proteinase K from the mold Tritirachium album Limber. Specificity and mode of action. 1) The specificity of proteinase K towards amino acid and oligopeptide nitroanilide substrates is investigated. 2) The active center of the enzyme contains an extended binding region consisting of several subsites. An integral part of the S1-subsite are hydrophobic areas which were investigated by systematic elongation of the carbon skeleton in carboxylic acid 4-nitrophenyl esters. On the basis of these studies, a possible model of the S1-binding site is proposed. 3) Kinetic parameters for the hydrolysis of substituted phenyl acetates catalyzed by proteinase K have been measured at pH 7 and 25 degrees C. Deacylation of an acyl-enzyme intermediate is probably the rate-limiting step. Acylation (kcat/km used as a measure) is modestly sensitive to the sigma values of the substituents (p = 1.33, r = 0.9108), indicating electrophilic assistance by the enzyme in the catalytic mechanism. 4) Hydrophobic forces apparently are not involved in the binding of the leaving group.


Preparation of a crystalline A1-(2-nitro-4-trimethylammoniophenyl) derivative of bovine insulin. The reaction of bovine insulin with the hydrophilic reagent 1-fluoro-2-nitro-4-trimethylammoniobenzene iodide at pH 8.2 has been reexamined. From the mixture of products a crystalline derivative substituted at the GlyAl terminus has been isolated. It has been characterized by amino acid analysis, end group analysis, cellulose acetate electrophoresis and sulphitolysis. The derivative has a biological activity (mouse convulsion assay) of 60% relative to insulin.


On the mechanism and stereochemistry of the malate-lactate fermentation of Leuconostoc mesenteroides. During the transformation of (2S, 3R) [3-3H]malate to (S) lactate no tritium exchange takes place. The stereochemical course of the decarboxylation studied with (2S, 3R) [3-2H]-malate in 3HOH/H2O and (2S, 3R) [3-3H]malate in 2H2O occurs with retention and is therefore the same as that determined by other authors for malic enzyme from vertebrates and from Escherichia coli. The malate-lactate fermentation is a useful procedure to prepare chiral methyl groups on a preparative scale starting from (2S, 3R) [3-H]malate.


Effect of oxygen shortage on the metabolism of oestrone in the hemoglobin-free perfused rat liver. The isolated rat liver was perfused with a haemoglobin-free, albumin containing salt solution at 28 degrees and 37 degrees C, respectively, with [4-14C]oestrone as substrate. During perfusion, the functional state of the liver was checked by continuously measuring the oxygen pressure and hydrogen ion concentration in the perfusion medium, flow rate, oxygen tension at various areas of the liver surface, and oxygen consumption; in addition, the following biochemical parameters were determined: ATP, ADP, lactate and pyruvate in liver tissue, and lactate and pyruvate in the perfusion medium. After 30 min of perfusion, free steroids, steroid glucuronides, steroid sulphates and the remaining water-soluble metabolites, present in liver tissue, perfusion medium and bile, were separated from each other and characterised by thin-layer and paper chromatography. It was found that, during perfusion at 37 degrees C, less hydroxylated metabolites were formed than at 28 degrees C. In contrast, metabolites whose formation is not directly oxygen-dependent, such as glucuronides and sulphates, arose in higher amounts at 37 degrees C than at 28 degrees C. It may be concluded that the relative shortage of oxygen at 37 degrees C leads to a selective impairment of metabolic pathways requiring a sufficient supply of molecular oxygen. Since oxidative processes play an important role in the biogenesis and metabolism of steroid hormones, it is evident that results, obtained in perfusion experiments with haemoglobin-free media at 37 degrees C, must be treated with reserve.


[A micro method for the determination of the configuration of histidine in peptides: evidence for partial racemization during peptide synthesis (author's transl)]. A gas chromatographic method for the determination of small portions of D-histidine together with an excess of L-histidine (or vice versa) is described. Histidine is degraded to aspartic acid by reaction with ozone. The enantiomers are esterified with (+)-3-methyl-2-butanol. After N-trifluoroacetylation the volatile diastereoisomers are separated in glass capillaries coated with a chiral stationary phase and determined quantitatively. For precise evaluations the partial racemization during peptide hydrolysis must be accounted for. The method was applied to some histidine peptides and about 5% racemization was measured.


Inhibitors of acrosin and granulocyte proteinases from human genital tract secretions. Human seminal plasma contains two acid-stable proteinase inhibitors, HUSI-II (Mr approximately 6500) and HUSI-I, (Mr approximately 11 000) with different inhibition specificities. The inhibitory activity of HUSI-II is strongly limited to trypsin and acrosin; both enzyme-inhibitor complexes are very stable (e.g. bovine trypsin-HUSI-II complex: Ki = 1 x 10(-10)M; human acrosin-HUSI-II complex: Ki = 2.7 x 10(-10)M). The inhibitor from human seminal plasma HUSI-II may therefore be seen as the natural antagonist of the sperm protease acrosin. In addition to pancreatic trypsin and alpha-chymotrypsin, HUSI-I forms strong complexes with neutral proteases of the lysosome-like granules from human granulocytes, for example, the elastase (Ki = 2.5 x 10(-9)M) and cathepsin G, the chymotrypsin like protease (Ki = 7 x 10(-8)M).


Preparation of a crystalline tris-(N-methylpyridinium)-derivative of bovine insulin. The reaction between 2-chloro-N-methylpyridinium iodide and bovine insulin at pH 8.5 afforded a trisubstituted derivative as main product. Following purification on DEAE-Sephadex A-25, this derivative crystallized from a citrate buffer in the form of large rhombohedra very similar to native insulin. This observation was confirmed by zero-layer X-ray precession photography. Amino acid analysis indicated substitution at the B1-terminus and on two of the four tyrosine residues. In the mouse convulsion assay the derivative showed no biological activity.


[(A1-beta-Alanine) insulin].

[(A1-beta-Alanine) insulin]. Starting from porcine insulin, A1-glycine was substituted by beta-alanine. The blood sugar lowering effect of the new analogue in the rabbit is about 45% of that of insulin. The half-maximal binding to partially purified rat liver receptors is about 46%, to transformed human lymphocytes about 54%, compared to insulin.


Analysis of velocity sedimentation techniques in cell separation. Influence of apparative and sample properties on separative power, resolution and sensitivity. In the present investigation the velocity sedimentation technique was analysed with respect to separation resolution, power and sensitivity. It was found that apparative modifications do not influence the resolution, which is a function of the contribution of apparative errors to the dispersion. A surprisingly small parameter of 0.15 was determined and it seems unlikely that this value can be improved. On the other hand an apparative modification is presented which improves the separation power and makes sample loading independent of the gradient filling. If cells (from rat bone marrow) were separated, a several times higher dispersion for a given cell volume was observed than was due to the apparative error. It was concluded that density variations were the major source of this dispersion. Since cell volume and density apparently show independent variations within a biological cell population the cell density cannot be disregarded if velocity sedimentation profiles are discussed in physical terms as is often done.


Intestinal absorption of polyunsaturated phosphatidylcholine in the rat. The mechanism of intestinal absorption of polyunsaturated phosphatidylcholine in an oil medium was studied with 1,2-di-[9,10,12,13-3H4]linoleoyl-sn-glycero-3-phospho-[N-14CH3]-choline, 1-[1-14C]linoleoyl-2-[9,10,12,13-3H4]-linoleoyl- and 1-[9,10,12,13-3H4]linoleoyl-2-[1-14C]linoleoyl-sn-glycero-3-phosphocholine, especially with regard to the stability of the ester bonds in position 1 and 2 of the phospholipid molecule. The absorption rate, as measured by the disappearance from the gastro-intestinal tract, was comparatively rapid in the first 6 - 8 h, but then became considerably slower. After 24 h more than 90% of the applied radioactivity was absorbed from the intestinal tract. Respiratory 14CO2 from the degradation of the unsaturated acyl moiety in position 2 is produced much more rapidly than that from the acyl group attached to the 1-position of the glycerophosphocholine backbone. Analyses of the liver phosphatidylcholine by specific enzymatic hydrolysis with phospholipase A2, 6 h after the application, showed that in the isolated PC 9 times more labelled fatty acids from the original 1-position were present than from the corresponding 2-position. In rats with lymph fistula it was shown that more than 90% of the acyl moieties of the administered 1,2-di-[9,10,12,-13-3H4]linoleoyl-[N-14CH3] glycerophosphocholine was transported by the chylomicrons. About one half of the 14C choline radioactivity of the glycerophosphocholine backbone was found in the chylomicrons and the other half in the liver. The 3H radioactivity distribution in the chylomicrons amounted to 25% in the phosphatidylcholine fraction and 75% in the neutral lipids. Positional specific analyses of the phosphatidylcholine present in chylomicrons confirmed the fact that the 1-position remained practically intact while the 2-position underwent considerable exchange with unlabelled fatty acids. Analysis of the liver of the animals with lymph fistula indicated that it was practically free of the 3H radioactivity derived from the acyl moieties but contained a high percentage of the 14C radioactivity of the choline group. The methyl groups of choline were oxidized only to a very small extent. These results demonstrate that during the absorption process, about one half of the absorbed polyunsaturated phosphatidylcholine is hydrolyzed to 1-acyl-lysophosphatidylcholine and reacylated again to phosphatidylcholine upon entering the mucosa cell, while the other half is completely hydrolyzed to free fatty acids and glycerophosphocholine or its hydrolysis products. The fatty acids released are utilized for the reassembly of triacylglycerides and phosphatidylcholine found in the chylomicrons.


The aging psychiatric hospital: an approach to humanistic redesign. Since funds for the construction of new treatment facilities are scarce, outmoded wards must be redesigned to meet the current needs of users. The authors feel the redesign must integrate modern therapeutic concepts, humanistic patient requirements, and pragmatic budgetary limitations. They review current theory about the effects of the physical environment on behavior, and they discuss such questions as to what extent the need for security should govern ward design and how pleasant the environment should be. They also describe a specific redesign for a 32-bed psychiatric ward of a large federal hospital constructed in the early 1950s.


South Carolina's village system.

South Carolina's village system. The authors describe the architectural, design, and mental health treatment concepts underlying the village system, a group of regional inpatient mental health facilities planned in South Carolina. One of the villages is currently under construction and is expected to open early in 1977. In design it resembles a residential community with all the elements of a normal small town or neighborhood.


Designing environments for mentally retarded clients. An environment that stimulates the auditory, visual, and tactile senses increases appropriate behavior among mentally retarded clients and thereby reinforces their learning activities. The author describes several low-cost deisgn techniques for giving a more stimulating and personalized atmosphere to large, existing institutions; discusses design requirements for group homes for retarded and handicapped clients in the community; and summarizes design ideas to be incorporated in new facilities built for retarded clients. She describes the implementation of her design ideas on a unit for blind retarded clients at the Mansfield Training School in Connecticut.


Safety and environmental factors in accreditation of facilities. The standards developed in 1972 by the Accreditation Council for Psychiatric Facilities of the Joint Commission on Accreditation of Hospitals cover 28 components of mental health programs, including the environment and patient safety. The author discusses the standards related to those two areas as they are set forth in the council's Accreditation Manual for Psychiatric Facilities, and he describes the difficulties psychiatric hospitals have faced in complying with them.


Perceptual factors in the design of environments for the mentally ill. Administrators and others responsible for the design of environments for the mentally ill must be aware that what might be considered irrelevant minutiae of design can have traumatic effects on patients. Because of patients' heightened sensitivity to sensory stimuli and their lessened ability to filter out or adjust to the effects of such stimuli, they are much more likely to be affected by their surroundings than are healthier people, and their social relationships are correspondingly affected. The author describes several kinds of visual, auditory, olfactory, and other stimuli that lead to distorted perceptions of oneself and others and of time and space. He recommends not a bland and stimulus-free environment, but one whose elements are unobtrusive and unambiguous.


Designing a mental health center to replace a county hospital. An architectural team worked closely with a county building committee and the administrative staff of a Wisconsin county hospital to design a new active-treatment facility. The goals included changing the hospital's negative image in the community, fostering a close relationship between all programs and all groups of users, and encouraging inpatients to make the best possible use of various treatment areas. Planning was interrupted by a community-wide controversy over the location, and the facility was eventually built on a compromise site. The new facility, a community mental health center serving two counties, has an open, split-level design accommodating six inpatient programs and a day hospital. An informal evaluation of the environment was made a year after the center was occupied.


A house is a center is a house: participatory design for a community counseling center. Using an environmental-design concept called pattern language, an architectural design group worked with five mental health center staff members to locate, design, and move into a setting for a satellite counseling center. The group first identified environmental problems, especially the lack of privacy, that existed in the counselors' previous setting, a remodeled storefront. However, in their new quarters, a two-story house, staff decided against having private offices; they used the upstairs rooms for counseling and the downstairs for shared spaces and work stations--inexpensive, individual cabinets with desk space that were placed to allow privacy for the user, yet permitted the communication essential for work as a team. After 18 months of use, staff's and clients' reactions to the setting continued to be favorable.


The rights of staff in the treatment of the mentally ill. If we accept the premise that adequate health care is the right of every individual, it follows that the provider of health care must, as a basic right, have the opportunity and the conditions necessary to provide such care. The author defines and describes a set of staff rights that bear directly on the treatment process. Rights of staff include having sufficient resources to provide adequate care, participating in the allocation of resources and the setting of priorities, being accountable for clinical matters to the highest governing authority, having clinical practice reviewed by peers, and practicing without excessive and unnecessary regulation.


Discharges against medical advice from voluntary psychiatric units. About one in every six voluntary mental patients discharges himself against medical advice. But despite the high rate of AMA discharges, the authors found the literature on the subject to be sparse and of little help in explaining why patients leave the hospital against medical advice. They examine the approaches that have been used thus far to study the phenomenon. They also report the findings of their own study, which showed that patients discharged against medical advice had a poorer response to treatment than those discharged in the traditional manner. The study identified factors that contributed to the poor response and were related to the discharge.


A time-limited treatment program for children and their families. In December 1972 the family psychiatric unit opened at Royal Jubilee Hospital in Victoria for the treatment of children under age 15 with emotional disturbances or behavior problems. Each child is admitted to the unit on a predetermined date for a pre-established period of five weeks, during which time the child's family is also involved in treatment. After the child is released from the unit, therapy continues for an additional five weeks in the family's home and then contact is terminated. To date more than 350 children have been admitted to the unit; a second admission has been required in only 21 cases.


Organizational systems for change offer an alternative to unions. Hospital management must begin to face the implications of unionization more directly. Certain current practices of most hospitals tend to increase vulnerability to unionization; they must be evaluated and modified. Strategies and programs for modernizing personel practices, for developing better employee information systems, and for training and developing managers need to be developed in order to effectively inhibit the intervention of labor unions.


Immature teratoma of the ovary with a neural component ("solid" teratoma). A clinicopathologic study of 20 cases. Twenty cases of immature teratoma of the ovary with a neural component are analyzed. A plea is made for use of the nomenclature adopted from the new World Health Organization classification of ovarian tumors, the past confusion over terminology and histogenesis of this rare tumor is discussed. All the primary tumors in the present series contained at least some immature tissues (predominantly of neural origin) and were thus graded from 1 to 3 according to the criteria of Thurlbeck and Scully. No grade 0 tumors ("benign solid teratomas") were identified. We believe that thorough sectioning almost always insures the identification of immature elements. The prognosis was closely related to the histologic grade, but correlated poorly withthe clinical stage, the latter being influenced by the common finding (25 per cent of the cases in this series) of peritoneal implants composed exclusively of mature glial tissue, which is associated with a benign clinical evolution. This phenomenon of maturation or differentiation appears to be the rule rather than the exception in this tumor, since implants are usually of better or equal differentiation when compared with their primary tumors and older patients tend to have lower grade tumors than younger patients. Since the majority of patients with this tumor are young, primary surgical therapy should be conservative, unilateral salpingooophorectomy often being sufficient. Spontaneous or operative rupture of the tumor capsule carries an increased risk of subsequent dissemination. We have noted impressive clinical responses in patients with disseminated tumors of a high histologic grade after treatment with triple chemotherapy (vincristine, actinomycin D, and cyclophosphamide) but do not recommend adjuvant therapy in patients with only grade 0 implants.


Bronchopulmonary dysplasia: the pulmonary pathologic sequel of necrotizing bronchiolitis and pulmonary fibrosis. A light and electron microscopic study was carried out in 21 infants in whom the pathologic diagnosis of bronchopulmonary dysplasia had been made. All the infants except two had the respiratory distress syndrome at birth, and all 21 had been treated with respirator and oxygen therapy for various periods of time. The pathologic alterations observed in all the infants studied were primarily damage of the bronchial and bronchiolar ciliary apparatus and mucous membranes, severe necrotizing bronchiolitis, and marked bronchiolar and alveolar fibrosis. These changes were more pronounced in infants who survived the longest period of time. Such inflammatory and fibrotic changes are known to predispose to destruction of lung tissue, emphysema, and pulmonary hypertension. Six of these 21 infants developed symptoms and signs of cardiac atrial or ventricular stress, including cor pulmonale, prior to their demise. These infants were among those that survived the longest periods of time, had the longest exposure to supplemental oxygen, and showed histopathologically severe pulmonary fibrosis and emphysema.


The human yolk sac and yolk sac carcinoma. An ultrastructural study. The ultrastructure of the yolk sac of a 39 day old human embryo was studied. The subcellular organization was suggestive of a highly specialized absorptive function proceeding in an exocelomic-viteline direction. These findings, compatible with intense metabolic activity, are at variance with the concept of rapid involution of the yolk sac following completion of its hemopoietic and angiogenetic functions. The speculation is advanced that a potential avenue exists in the yolk sac whereby maternally derived products encounter fetal endoderm. Ultrastructural features in the normal yolk sac were compared to those existing in a tumor showing the "endodermal sinus" pattern, and reviewed in the light of the pertinent literature. These findings support the concept that attributes an endomesoblastic derivation to such neoplasms.


Pseudomembranous colitis associated with antibiotics. Colitis associated with antibiotics, particularly with lincomycin and clindamycin, is a well established entity. The colitis may be clinically and radiologically very difficult to distinguish from inflammatory bowel disease, including Crohn's disease and ulcerative colitis. A wide spectrum of pathological features is described with various antibiotics. However, the pathological picture in the pseudomembranous form is quite distinctive. The most important histological findings include a "mushroom-like" or "explosive" appearance of the pseudomembrane with a sudden transition to normal mucosa adjacent to the lesion. Rectal biopsy is both an accurate and a rapid method of establishing the diagnosis.


Embryonal rhabdomyosarcoma (sarcoma botryoides) of the common bile duct: a case report. A case of sarcoma botryoides of the common bile duct is described. A review of the literature reveals 24 reported cases of this entity. The clinical features of the disease and the poor prognosis associated with it are illustrated by a detailed presentation of the case. Although rare, this condition must be included in the differential diagnosis of jaundice in infancy or early childhood. Pathologic findings of diagnostic value are re-emphasized.


Veno-occlusive disease of the liver associated with oral contraceptives: case report and review of literature. This report concerns a young woman who, after taking a contraceptive preparation orally for three years, developed the Budd-Chiari syndrome as the result of a widespread chronic obliterative process involving the intrahepatic efferent venous system. Her prolonged course, which failed to respond to an end-to-side portacaval shunt procedure, mimicked chronic hepatitis evolving to cirrhosis. Additional noteworthy features were the presence of two small benign hepatic adenomas, observed both at biopsy and at necropsy, a lesion recently recognized as a complication of anovulatory drugs, and widespread hepatic calcifications found at autopsy.


Trisomy 13 mosaic presenting as cleft lip and palate. A mosaic trisomy 13 presenting as a case of cleft lip and palate in the newborn is described. However, when the child was admitted to hospital at the age of 6 weeks because of failure to gain weight and a malformation of the great vessels was demonstrated, cytogenetic studies were carried out. The diagnosis of mosaic trisomy 13 (90% normal, 10% trisomic) was established from a leukocyte culture. Since, occasionally, mosaic trisomy 13 may mimic cleft lip and palate in the newborn, cytogenetic studies are indicated in the presence of any additional anomaly.


A study of several genetic biochemical markers in Sherpas with description of some variant phenotypes. Gene frequencies for 23 genetic biochemical markers have been determined in one sample of Sherpas and two smaller samples, one of Tibetans living in Nepal and one of 'mixed' Nepalese. Sherpas presented a high GPT1 (0.71) and an appreciable incidence of HbE (about 2%). Variant phenotypes for PGM1, PHI, PepB and C were observed.


Serum cholinesterase variants in African leprosy patients resident in Rhodesia. Blood samples from 580 African leprosy patients living in Rhodesia have been phenotyped for the plasma cholinesterase variants. The Africans have been grouped according to country of origin and tribal affiliation. We have found no individual with an Ea1 gene and are unable to resolve the contradictory evidence for an association between this gene and leprosy. The frequency of the Ef1 gene is higher than that usually found in Caucasian populations, being 0.046 in lepromatous leprosy patients and similar to the 0.056 found in healthy African controls. In tuberculoid leprosy patients the frequency is, however, significantly lower at 0.019. On the other hand, the frequency of the C5+ variant is essentially the same for the tuberculoid leprosy patients and the healthy controls (4%) while for the lepromatous leprosy patients it is about 7% approaching the 10-15% found in many Caucasian populations.


The cholinesterase variants found in some African tribes living in Rhodesia. Blood samples from 1,614 Africans living in Rhodesia have been phenotyped for the cholinesterase variants at the E1 and E2 loci. 24% of the African population were non-Rhodesian by birth. 1,227 Rhodesians aligned themselves to 20 tribes, 191 Malawians to 8 tribes. 162 Mozambique Africans to 9 tribes and 34 Zambians to 8 tribes. A high frequency of 0.036 for the Ef1 gene, which varies from tribe to tribe, has been found in Rhodesian and Malawian Africans. Similar high frequencies for this gene are recorded for Zambian (0.045) and Mozambique Africans (0.034). The frequencies of the Es1 gene in these groups are 0.013 (Rhodesian), 0.009 (Malawian), and 0.016 (Mozambique African). The small Zambian sample showed evidence for neither the Es1 nor the C5+ electrophoretic variant. The absence of the Ea1 gene in the 1,613 Africans provides additional evidence of the rarity of this gene in negroid populations. The frequency of the C5+ variant in Rhodesian, Malawian and Mozambique Africans, although varying from tribe to tribe within the range of 0-8%, averages 3% in each group. These represent low frequencies for this variant when compared to other populations. No rare or 'private' electrophoretic variant has been found.


Esterase D and superoxide dismutase polymorphisms in Iraq. From a study of 320 individuals superoxide dismutase (SOD) variants are shown to be present in Iraq, and it is suggested that this genetic system may be useful for the study of historical population movement in the Middle East. The esterase D gene frequencies show no significant heterogeneity among the different regions of the country.


Preparation of delipidized serum protein for use in cell culture systems. A rapid procedure for the preparation of delipidized serum protein is described. The delipidized protein can be used for the maintenance and growth of tissue culture cells in a lipid-free environment. The extraction procedure greatly reduces all serum lipid classes and the delipidized protein supports the growth of a variety of cells in culture.


The measurement of spatial precursor distributions in cell culture. The distribution of incorporated radioactive precursors, for both DNA and protein synthesis, has been measured with a resolution of about 1 mm in cell cultures, using a scanning technique. Either gamma radiation and X-rays or beta radiation (electrons) were detected by scintillation detectors. Spectrophotometer measurements with a resolution of 1 mm gave good estimates of cell density changes. Glioma cell colonies were used to compare this technique with autoradiography. Variables such as the density of labelled cells and percentage of labelled cells could be estimated rather accurately. For example, an increased cell density was correlated to a local decrease in DNA synthesis.


Effect of auxin-cytokinin interaction on organogenesis in haploid callus of Pelargonium hortorum. Bud differentiation from haploid anther callus of geranium was achieved on Murashige and Skoog medium supplemented with 0.5 mg per 1 NAA and 2.5 mg per 1 kinetin. At concentrations higher than 5 mg per 1 kinetin, malformation and tissue senescence were evident.


A chemotactic receptor for val(ala)-gly-ser-glu on human. Preferential eosinophil chemotactic activity is an in vitro and in vivo property of eosinophil chemotactic factor of anaphylaxis (ECF-A), a mixture of two peptides, Val-Gly-Ser-Glu and Ala-Gly-Ser-Glu, isolated from extracts and anaphylactic diffusates of human lung tissue. Purified native and synthetic ECF-A share with the synthetic N-formyl methionyl peptides such features as in vitro activity in nanomolar amounts, high dose inhibition of effect and a requirement for hydrophobic amino acid residues. The capacity of the substituents of ECF-A, Val-Gly-Ser, Ala-Gly-Ser, and Gly-Ser-Glu to modulate eosinophil chemotaxis has permitted a preliminary functional characterization of an eosinophil surface receptor. The activity, specificity, and structural characteristics of the active tetrapeptides suggest that distinct interactions of the peptide with a stereospecific receptor on the eosinophil surface is required for chemotactic movement.


Receptor mobility and its cooperative restriction by ligands. The functional significance of membrane fluidity and receptor mobility in lymphoid cells has been studied in the recent literature. Although far from clarified, the role of membrane fluidity in achieving control over cell activity is probably important; it allows cooperative interactions over long distances. Here, the emphasis is put on the phenomenon of restriction of receptor mobility by ligands such as Concanavalin A, a phenomenon discovered in recent years using morphological techniques. We discuss in some detail our own approach for studying this phenomenon. This consists of quantitatively measuring the active sites on cell-bound lectin molecules by subsequent fixation of horse-radish peroxidase. This study has shown a cooperative binding of Concanavalin A to cells which corresponds to a modification of the membrane, leading to the recruitment of new receptors. The existence of a post-binding event, that we have called micro-redistribution, has been shown at 4 degrees C, through the use of peroxidase binding to cell-bound lectin. A cooperative restriction of receptor microredistribution is observed when the cooperative recruitment of receptors induced by increasing concentrations of Concanavalin A occurs. Both phenomena were shown to be modulated by drugs such as colchicine and cytochalasin B. The characteristics of this modulation suggest that density and distribution of receptors are dependent upon the state of a multimeric submembrane structure which is still functional at 4 degrees C.


Binding properties of bovine colostral anti-dinitrophenyl (dnp) immunoglobulins g2 (igg2). The number of combining sites per mole of bovine colostral anti-DNP IgG2 was found to be 2 and the total affinity constant was 0.81 X 10(4) M-1. Unlike bovine colostral IgG1, the nonspecific binding of H3-epsilon-DNP-1-lysine by IgG2 as a function of increased concentrations, did not show a negative, cooperative slope. Spectral measurements made with anti-DNP IgG2 in the reference cell vs. anti-DNP IgG2 plus hapten in the experimental cell revealed a hypochromic, red shift from 363 to 365 nm. If the reference cell contained hapten, a red shift from 280 to 283 nm and an enhancement in the extinction coefficient of IgG2 was observed. These spectral changes were not observed if nonspecific IgG2 was substituted for anti-DNP IgG2 in the experimental cell. The enhancement in the extinction coefficient was interpreted to be due to a possible exposure of previously buried tryosine and tryptophan residues.


Semi-micro quantitative gel precipitin reaction. Quantitative gel precipitin reactions were made on a semi-micro-scale. This required overall reduction of the dimensions of the necessary apparatus. Resulting from this was the ability to use agarose gel at low concentration not possible with the conventional macro quantitative gel precipitin technique. Further advantages were the reduction of time for the completion of the experiments and also that of the volume of the reactants. Diffusion coefficients of antigens determined by the semi-micro technique agreed with those obtained with the more accurate optical methods.


Thymosin induced changes in the cell cycle of lymphocytes from aging neonatally thymectomized rats. The potential role of the endocrine thymus on the cell cycle of lymphocytes from aging neonatally thymectomized rats (LAT cells) was investigated using thymosin fraction 5. Thymosin rendered LAT cells similar to normal lymphocytes with regard to reconstituting the median S+G2 phase of the LAT cell. This is the first study to demonstrate an effect of thymosin on the cell cycle of lymphocyte populations.


The class of antibody involved in the antibody-dependent cell-mediated lysis of chicken erythrocytes. Affinity chromatographic experiments demonstrated that IgG was the class of antibody involved in the in vitro lysis of chicken erythrocytes by antibody and normal mouse effector cells. Further experiments designed to investigate the subclass of IgG involved, indicated that IgG2a was active and provided further evidence for the inactivity of IgM, either in the native 19S or chemically degraded 7S form. From the results obtained, the possibility of ADCC-inducing activity in the other IgG subclasses could not be excluded.


Stimulating activity of Brucella fractions in a human lymphocyte transformation test. Correlation with humoral and cellular immunity. 'PI' a phenol-extracted fraction from Brucella melitensis has been used for years in our laboratory for the vaccination of laboratory and agricultural workers considered 'at risk'. In the present work it has been used in vitro, together with another phenol-extracted and peptidoglycan containing fraction (4A) in a lymphocyte transformation test. Correlation between stimulation index, humoral antibodies to Brucella and the cutaneous response to PI have been studied in subjects vaccinated with the PI fraction, in Brucellosis patients and normal controls.


Lymphocyte emigration from high endothelial venules in rat lymph nodes. Sequential events during lymphocyte emigration from high endothelial venuses (HEV) were studied by scanning and transmission electron microscopy combined with regional perfusion techniques. The results indicate that blood lymphocytes selectively adhere to HEV surfaces through microvilli which attach to shallow pits on the luminal surfaces of high endothelial cells. These intercellular contact points resist hydrodynamic and osmotic shearing forces, but can be disrupted by treatments which remove endothelial glycocalyx, hydrolyse lymphocyte surface glycoproteins, or chelate divalent cations. After this initial attachment phase, lymphocytes enter apical clefts between endothelial cells where they assume a motile configuration characterized by loss of microvilli and formation of irregular surface folds. Intramural lymphocytes adhere to adjacent endothelial cells through macular and villous contacts. Fibrillar electron-dense material traverses the 15-20 nm gap at these points of adhesion. Microtubules and microfilaments are also seen around areas of cytoplasmic constriction in these motile lymphocytes. The migrating lymphocytes show cytoplasmic polarity which is oriented in the direction of movement as they pass through extracellular spaces in the venular wall and cross successive laminations in the perivascular sheath to enter the node. Since these lymphocytes enter channels between endothelial cells which are stained by intralymphatic injections with horseradish peroxidase, it is suggested that their entry into the node depends upon migration along a chemotactic gradient.


Effects of activated complement components on enzyme secretion by macrophages. Purified cleavage products of the guinea-pig complement component C3, namely C3b and C3a, interact with guinea-pig and mouse macrophages in culture to induce a dose- and time dependent release of lysosmal enzymes into the medium. In the case of C3b the selectivity of the release of hydrolases, which occurs without cell killing, is shown by morphological observations and the failure of lactate dehydrogenase to appear in the medium. However, lysosomal enzyme release in the presence of C3a is accompanied by loss of cellular lactate dehydrogenase. Preincubation of C3b with anti-C3 Fab inhibits its attachment to macrophages, after which there is hardly detectable enzyme release into the medium. We have found that stimulated macrophages release enzyme(s) which can cleave C3, generating more C3b either directly or via the alternative pathway; the C3b so formed would induce further enzyme release. This amplification system may provide an explanation for the ability of macrophages to generate mediators of inflammation and cause tissue damage and degradation at sites of chronic inflammation while retaining their ability for long periods of time.


Formation in the presence of C3 nephritic factor (C3NeF) of an alternative pathway C3 convertase containing uncleaved B. C3 nephritic factor (C3NeF) interacts with native C3 and B in the absence of D to generate a C3 convertase containing an uncleaved form of B. Dose response studies with C3NeF and B, respectively, revealed incremental C3 inactivation without loss of B. These findings are in agreement with the previous isolation from such reaction mixtures of a 10S complex containing haemolytically inactive C3 and active B and manifesting C3 convertase activity. Functional contamination of C3 with C3b was negated by demonstrating that pretreatment of C3 with C3bINA had no effect on its subsequent interaction with B and C3NeF to generate C3 convertase activity, while pretreatment of C3b eliminated its effective interaction with B and C3NeF. Relatively higher concentrations of C3bINA present during interaction of C3, B and C3NeF suppressed C3 inactivation, indicating its dependence on amplification by utilization of the initial C3b generated. Trace quantities of D were not found by functional analyses of C3, B and C3NeF and pretreatment of these proteins with a concentration of DFP sufficient to suppress D activity had no effect on their effective interaction. The introduction of D to mixtures of C3NeF, B, and C3 resulted in B clevage and more efficient expression of C3 convertase function as defined by a reduced requirement for C3NeF.


Eosinophils and mediators of anaphylaxis. Histamine and imidazole acetic acid as chemotactic agents for human eosinophil leucocytes. Histamine and one of its major catabolites, imidazole acetic acid (ImAA), were selectively chemotactic for human eosinophils, whereas L-histidine and other histamine catabolites including 1,4-methylhistamine, 1-methyl-4-imidazole acetic acid and N-acetylhistamine were inactive in eosinophilotaxis over a large dose range. The dose response for histamine was dependent on the chemotaxis incubation time and the source of eosinophils, although the latter was not clearly associated with particular disease states. When histamine and ImAA were combined the chemotactic response was similar to that obtained when one agent was assayed alone, no additive or synergistic effects being observed. There was cross-deactivation between histamine and ImAA. These experiments suggest that histamine and ImAA activate the same chemotactic recognition mechanism for eosinophils. Thus ImAA joins histamine and the tetrapeptides (ECF-A) as anaphylaxis-associated selective chemoattractants for human eosinophils


Lymphocyte nucleolar activation as a marker of autoimmune disorders. II. Observations in patients with systemic lupus erythematosus and other connective tissue diseases. An increased number of lymphocytes with active nucleoli was found in the peripheral blood of most patients suffering from connective tissue diseases associated with autoimmunity.


Effects of Corynebacterium parvum treatment and Toxoplasma gondii infection on macrophage-mediated cytostasis of tumour target cells. Injection of mice with Corynebacterium parvum or living or killed Toxoplasma gondii was studied to determine the efficacy of these treatments in activating peritoneal macrophages to inhibit the uptake of [3H]TdR (cytostasis) by tumour target cells in vitro. In the presence of activated macrophages from mice treated i.p. with a wide dose range of either C. parvum or living Toxoplasma, cytostasis was usually greater than 99%. This population of activated macrophages was transient in C. parvum-treated mice, but persists, probably for life, in Toxoplasma-infected mice. Whereas the i.p. route of administration of C. parvum was more efficient in activating macrophages than the i.v. route, the s.c. route appeared to be relatively ineffective. Treatment with killed Toxoplasma by any route was also relatively ineffective in activating macrophages. In contrast Toxoplasma infection resulted in highly activated peritoneal macrophages, regardless of the route of administration. Depending upon the route of initial treatment, the route of readministration of C. parvum had either no appreciable effect or resulted in a marked alteration in the cytostatic capacity of peritoneal macrophages.


EA rosette-forming lymphoid cells in chickens: specificity of the Fc receptor and its relationship to other surface antigens. The receptor for erythrocyte-antibody (EA) complexes on the surface of chicken lymphoid cells was investigated using a rosette assay. The chicken EA receptor binds chicken immunoglobulin of the IgG class but not the IgM class. Binding to the EA receptor is dependent upon the Fc region of the immunoglobulin. No receptor for complement analogous to the mammalian C3b receptor was demonstrated on chicken lymphoid cells using the rosette assay. Inhibition studies utilizing immunoglobulins from several species demonstrated that chicken spleen cells do not bind mammalian immunoglobulin but may bind immunoglobulin of other avian species (turkey and duck) and a reptilian species (turtle). The chicken EA receptor is distinct from cell membrane bound immunoglobulin light chains, bursa-specific antigens and thymus-specific antigens. The receptor for EA complexes on chicken lymphoid cells is compared with the Fc receptor on mammalian lymphoid cells in the light of these observations.


In vitro synthesis of beta1C/beta1A globulin (the C3 component of complement) by tissues and leucocytes of mice. The synthesis of beta1C/beta1A globulin (the C3 component of complement) was studied in in vitro cultures of mouse tissues and leucocytes. Often culture in medium containing 14C-labelled amino acids, the newly synthesized radioactively-labelled protein was identified by radioimmunoelectrophoresis. Synthesis was found in spleen, liver, trachea, lung and Peyer's patch tissue, but was only occasionally detectable in cultures of intestine. Cultures of the glass-adherent cells from bone marrow, blood, lung washings and subcutaneous tissue all showed strong synthesis, indicating that the ability to synthesize beta1C/beta1A is a property shared by mononuclear phagocytes from many sites. In contrast, the nonglass adherent cells showed no synthesis of beta1C/beta1A, although these cells were shown to be viable by their ability to synthesize immunoglobulins.


Kinetics and localization of IgE tetanus antibody response in mice immunized by the intratracheal, intraperitoneal and subcutaneous routes. The heterologous adoptive cutaneous anaphylaxis system was used to determine the kinetics of appearance of IgE-producing cells in various lymphoid tissues of mice following intratracheal (i.t.), intraperitoneal (i.p.), or subcutaneous (s.c.) immunization with tetanus toxoid and Bordetella pertussis organisms. Immunization, i.t. and i.p., produced similar patterns of response with the bronchial lymph nodes quantitatively exceeding the responses in other lymphoid tissues. In both cases the splenic lymphocyte response was second only to the bronchial and both appeared to parallel the serum PCA antibody. It is suggested that both responses represent draining lymph node responses since the bronchial lymph node drains both sites of immunization. After s.c. immunization a primary response of low order was found in the draining popliteal lymph node but not elsewhere. Although a dissociation was seen between responses obtained in various lymphoid tissues following s.c. and i.p. or i.t. immunization, no real evidence for a local mucosal response, such as has been reported for IgA, was obtained. These results lend experimental support to the observations that intratracheal and intraperitoneal immunization routes are most effective in production of IgE antibodies.


The effects of nutritional rehabilitation on antibody production in protein-deficient mice. The transfer of chronically protein-deficient mice to an optimal diet a few days before immunization with sheep erythrocytes, tetanus toxoid or Brucella abortus vaccine either failed to increase antibody production above the level produced by deficient mice, or suppressed the responses to below those produced by deficient mice or normally-fed controls. Transfer to high protein diet on the day of immunization or feeding deficient mice the normal diet for just 2 days at the time of injection produced higher titres than did transfer a few days before immunization. Secondary responses to TT were affected by transfer to the normal diet at priming, rather than at rechallenge. Some mechanisms which may explain these findings and their implications for immunization schedules in malnourished humans are discussed.


Purification of cobra venom factor from phospholipase A contaminant. It has been demonstrated that cobra venom factor prepared by the usual combination of ion exchange chromatography and sephadex gel filtration is contaminated by substantial amounts of a 'heavy' phospholipase A. The two activities may be separated by isoelectric focusing. Cobra venom factor focuses at pH between 5-75 and 6-75 whereas the phospholipase is all found at pH below 7-75. In certain test systems, particularly in vitro, and particularly where albumin concentrations are low, the contaminating phospholipase may produce effects that have been attributed to complement activation.


Pinocytic rates of macrophages from mice immunized against Toxoplasma gondii and macrophages stimulated to inhibit toxoplasma in vitro. The rate of pinocytosis by macrophages when measured by uptake of horseradish peroxidase was significantly reduced during toxoplasma infection of the cells in vitro when the macrophages were from toxoplasma-immune mice and when control cells were stimulated in vitro to inhibit toxoplasma multiplication. There was, however, no direct correlation between reduced pinocytosis in this model and inhibition or enhancement of toxoplasma multiplication. We conclude that a reduced pinocytic rate is a feature of the unstimulated toxoplasma-immune macrophage, but this change in rate alone does not correlate with the cell's ability to inhibit toxoplasma. In addition, we observed that enhanced pinocytosis as seen in the elicited macrophage was not a requirement for inhibition of toxoplasma multiplication.


Lymphocyte and macrophage responses after vaccinia virus infections. Using a semimicromethod with washed whole blood, in vitro lymphocyte responses of rabbits to intradermal infection with vaccinia virus was studied. Peritoneal exudate macrophages were infected with vaccinia in vitro to determine the time of appearance of activated macrophages. After primary infection, an increase in spontaneous incorporation of thymidine by blood cultures was found as early as 2 days postinfection. This effect was at a maximum at 7 to 10 days, with counts up to 100-fold higher than before infection. Incubation of these cultures with concanavalin A showed a marked decrease in stimulation index as compared with normals. Although only a transient stimulation with vaccinia was found during the acute infection, stimulation indexes of 2 to 3 were obtained during convalescence. Macrophages from rabbits early after infection supported vaccinia replication, whereas those at day 6 or later resisted infection. Macrophage resistance persisted for 2 to 3 weeks. The response of lymphocytes from rabbits reinfected with vaccinia after 15 weeks differed, with a small increase in spontaneous thymidine uptake, a smaller depression in concanavalin A stimulation, and a greater specific response to vaccinia. Macrophage activation occurred earlier and persisted for a longer time after secondary infection.


Immunological control mechanism against cholera toxin: interference with toxin binding to intestinal receptors. The immunological control mechanism against cholera toxin (CT) in the small intestine of rats was studied in vivo. CT binding to intestinal receptors was determined by injected radiolabeled CT into the loops of rat small intestine and subsequently separating purified microvillus membranes from mucosal scrapings of those loops. substantial radioactivity (10(5) cpm/mg of microvillus protein) was present in microvillus fractions of small intestine exposed to 125I-labeled CT compared to radioactivity (10(2) cpm/mg) in fractions from intestine exposed to radiolabeled bovine serum albumin (BSA) used as a control. CT binding to intestinal receptors was significantly inhibited (P less than 0.02) in rats immunized with crude toxin by a combined intraperitoneal and oral method compared to CT binding in animals immunized with BSA or controls, suggesting a specific relationship between intestinal antitoxin and inhibition of binding. Furthermore, ligated ileal loops from CT-immunized animals showed a significant decrease in fluid accumulation when exposed to CT compared to loops from control or BSA-immunized animals, suggesting that antitoxins also interfered with the biological action of CT under conditions of immunization. These studies provide direct evidence that intestinal antitoxins protect against CT-induced diarrhea by interfering with the attachment of the toxin to the intestinal microvillus surface.


Fate of rubella genome ribonucleic acid after immune and nonimmune virolysis in the presence of ribonuclease. To determine whether rubella virion ribonucleic acid (RNA) becomes accessible to nuclease attack after immune lysis of the viral envelope, virions containing radioactively labeled RNA were examined in three ways with the following results. (i) Incubation of purified virus with heat-inactivated rubella convalescent human serum and guinea pig complement resulted in an increase in acid-soluble RNA. Antibody was required; the reaction was temperature dependent and was blocked by ethylenediaminetetraacetic acid. When exogenous nuclease was added prior to lysis, radioactivity in virions was reduced to 15% of that in unlysed control pellets (ii) Sucrose gradient sedimentation profiles of RNA released from lysed and unlysed virions under controlled conditions showed that the nuclease content of serum-virus mixtures was sufficient to eliminate all RNA of genome size, although degradation was not complete. (iii) Virions were also lysed by unheated human immune sera in the absence of guinea pig complement and by some, but not all, unheated antibody-negative sera.


Passive hemolysis test for antibody to Treponema hyodysenteriae. Swine were infected orally with pure cultures of Treponema hyodysenteriae, and the humoral antibody response was measured by a passive hemolysis test (passive hemagglutination test with the use of complement). Antibody to T. hyodysenteriae was detected as early as 1 week and later on at 4 months after exposure. However, peak serum titers were obtained after challenge at 4 weeks postinfection. The usefulness of the test as a potential diagnostic tool for antibody to T. hyodysenteriae is discussed.


Model delineating the effects of a salivary pellicle on the adsorption of Streptococcus miteor onto hydroxyapatite. A model describing the adsorption isotherms for Streptococcus miteor strain 26 to untreated and saliva-treated hydroxyapatite was developed. The strengths of the adsorption bonds in the two systems were similar, however, the presence of selectively adsorbed salivary components increased the number of available binding sites fourfold.


Amplified migration inhibition effect.

Amplified migration inhibition effect. Upon exposure to specific antigen in tissue culture, sensitive lymphocytes released macrophage migration inhibition factor and other lymphokines into the supernatant culture medium. Migration of peritoneal macrophages from nonsensitive animals was inhibited in the presence of such supernatants. However, with previous techniques it was found that an inhibitory effect was present at only low low titers (less than 10(2)). It is therfore of great interest that by increasing cellular density, the total number of cells being kept constant, inhibitory activity can be amplified by a factor as great as 10(10). This amplification was observed only when lymphocytes and macrophages were loosely packed, as by spontaneous sedimentation in a conical test tube. The effect was abolished by dispersing the cell suspension in a flat-bottomed flask or, alternatively, by shaking the test tube so that intimate prolonged intercellular contact was prevented.


Attempts to serologically classify feline caliciviruses on a national and an international basis. A serological classification of five feline calicivirus (FCV) strains of Swiss origin and 13 isolates of Austrian origin was attempted. The antisera used had been prepared in rabbits against the five Swiss strains and in goats against six American strains. Homologous and heterologous neutralization tests were made in tube cultures using sera at dilutions that contained 20 antibody units, in unabsorbed sera at low dilution, and in sera after subjection to three consecutive absorptions with lyophilized feline liver powder. None of these procedures resulted in the delineation of serotypes. A study using 20 antibody units yeilded FCV strains with broad-spectrum antigenicity, which seem promising for vaccination trials and for covering global needs. In addition, readily neutralized strains suitable for epidemiological work on cat sera, and possibly also for measuring humoral response to vaccines, could be recognized. The extent of serum titer variance, after 10-fold variance of virus input, was established as being only two- to threefold. Five Swiss FCV strains have been shown to remain antigenically stable over 10 years of laboratory passage. FCV strains and the procedure using 20 antibody units offered a suitable approach for international comparative work. FCV strains with little cross-neutralization should be subjected to higher antibody concentrations of existing antisera before attempting to create serotypes. Liver powder absorptions, which reduced homologous and heterologous neutralization titers to similar extents, subsequently proved to be unsuitable for use in FCV classification.


Evidence for two immune inhibitors from Bacillus thuringiensis interfering with the humoral defense system of saturniid pupae. Mutants of Bacillus thuringiensis lacking either beta-exotoxin or gamma-endotoxin were compared for their virulence using pupae of a giant silk moth. Known doses of viable log-phase bacteria were injected, and the response was followed as the number of viable bacteria in the hemolymph. The results obtained imply that, in the system used, neither the beta-exotoxin nor the gamma-endotoxin and the sporeforming ability are of importance for virulence. Results with sterile culture filtrate from B. thuringiensis have given evidence for the production of two inhibitors, A and B, which interfere with the humoral defense system in pupae of Hyalophora cecropia. Inhibitor A, which blocked the lysis of Escherichia coli,was precipitated by trichloroacetic acid and sensitive to heating. Inhibitor B, which blocked the killing of Bacillus cereus, was soluble in trichloroacetic acid and resistant to 90 degrees C for 5 min. Both inhibitors are believed to contribute to the insecticidal nature of B. thuringiensis.


Inhibition of human neutrophil chemotaxis in vitro by phenothiazines and related compounds. Chlorpromazine (CPZ) and three other phenothiazines and the structurally related antidepressant drugs imipramine and amitriptyline were found to depress human neutrophil chemotactic responsiveness. A 7 X 10(-6)M solution of CPZ inhibited chemotaxis, whereas concentrations of the other tested drugs 10 to 1,000 times greater than this were needed to inhibit chemotaxis. This effect of CPZ could not, however, be demonstrated when testing neutrophils from patients treated with the drug. The inhibition of chemotaxis was reversible when CPZ-incubated neutrophils were washed before testing for chemotactic responsiveness. CPZ affects neutrophil funcjtion as well as other aspects of immune response.


Extrachromosomal elements in a variety of strains representing the Bacteroides fragilis group of organisms. Previous nucleic acid association studies have identified at least nine deoxyribonucleic acid (DNA) homology classes of the Bacteroides fragilis group of organisms. Using these classes as a taxonomic framework, we have screened representative strains of the B. fragilis group for the presence of extrachromosomal (plasmid) DNA. [3H]thymidine-labeled cell lysates were subjected to sodium dodecyl sulfate-salt precipitation, and supernatant fractions from such preparations were analyzed using cesium chloride-ethidium bromide equilibrium centrifugation. One strain from each group was examined in this fashion. Five of the strains were judged to contain no detectable plasmid DNA; however, four strains were observed to yield satellite bands corresponding to covalently closed circular plasmid DNA. Plasmid DNA from such gradients was subjected to velocity sedimentation through both neutral and alkaline sucrose gradients to determine molecular size. A 23 X 10(6)-molecular-weight plasmid was found in a B. fragilis strain representing one DNA homology group of this species, whereas a 3 X 10(6)-molecular-weight plasmid was found in a B. fragilis strain representing a second homology group. Similarly, a 31 X 10(6)-molecular-weight plasmid was found in a Bacteroides thetaiotaomicron strain representing one DNA homology group of this species, whereas a 3 X 10(6)-molecular-weight plasmid was found in a B. thetaiotaomicron strain representing a second homology group. In all instances, the small-molecular weight plasmids were present to the extent of about 15 copies per chromosomal equivalent, whereas the large plasmids were present to the extent of approximately 1 copy per chromosomal equivalent. The biological function of these plasmids is unknown.


Tumor inhibition in mice by lipopolysaccharide-induced peritoneal cells and an induced soluble factor. Lipopolysaccharide (LPS) was shown to prevent tumor growth in BALB/c mice when administered either prior to or after the inoculation of lethal doses of tumor cells. An attempt to elucidate the mechanism of this phenomenon utilizing in vivo protocols was made by the adoptive transfer of tumor protection with peritoneal cells as well as with cell-free peritoneal fluids obtained from non-tumor-bearing, LPS-stimulated syngeneic mice. The in vivo-activated peritoneal cells from LPS-treated mice were capable of adoptively transferring tumor protection at peritoneal cell to tumor cell ratios ranging from 1,000:1 to 100:1. Experiments were also performed that indicate that: (i) LPS exerts no direct toxic or inhibitory effect on the tumor cells, and (ii) that residual LPS present in cell and fluid preparations was not responsible for such protection.


Lymphocyte function in experimental african trypanosomiasis: mitogenic effects of trypanosome extracts in vitro. Extracts of Trypanosoma brucei and Trypanosoma congolense were incubated in vitro with nonimmune lymphocytes of mice, rats, guinea pigs, and rabbits in order to test for mitogenic effects or for other characteristics of polyclonal B lymphocyte activators. Trypanosome extracts (TE) were not mitogenic for spleen cells of mice, rats, and guinea pigs in vitro, nor did the parasite extracts alter the mitogenic responses of lymphocytes from these animals to known B- and T-cell mitogens. TE also failed to induce polyclonal antibody synthesis in mouse spleen cell cultures in an in vitro antibody response system, in contrast to the effects of bacterial lipopolysaccharide, a known polyclonal B cell activator. Rabbit spleen cell and peripheral blood lymphocyte cultures, however, were stimulated by TE to undergo blastogenesis in vitro. Incubation of rabbit lymphocytes with phytohemagglutinin (PHA) and TE or anti-rabbit immunoglobulin serum and TE revealed an additive effect only in terms of the TE-plus-PHA culture responses; these findings suggest that a non-PHA responsive lymphocyte population, possibly B lymphocytes, is stimulated by TE in rabbits. The relationship of trypanosome-induced lymphocyte mitogenic stimulation to other immunological dysfunctions occurring in chronic African trypanosomiasis is discussed.


Host immune potentiation of drug responses to a murine mammary adenocarcinoma. II. Effect of melphalan therapy on the host immune system. The role of chemotherapy in influencing tumor-specific immunity to a mouse mammary adenocarcinoma was investigated. By studying different stages of tumor growth we were able to identify several factors important to drug-induced tumor regression: (1) antibody response, (2) delayed hypersensitivity, (3) sensitivity of tumor cells to immune attack and (4) tumor burden. The presence of tumor-specific delayed hypersensitivity and circulating antibody as well as specifically armed monocytes in the tumor mass characterize the T1699 adenocarcinoma. Successful chemotherapy had previously been shown to depend on prior establishment of the above immune responses. Treatment with alkylating agents was marked in all animals by abrogation of a humoral response to the tumor when drug was given early (day 7), and was associated with poor chemotherapeutic results. Later treatment (day 10) was associated with depression of antibody titers only in the minority of animals not responding to drug and prolongation of the delayed hypersensitivity response in all treated animals. Tumors recurring following initial drug-induced regression were marked by lack of delayed hypersensitivity in the host, lack of drug response and suppression of humoral immunity following treatment. Successive passage of cells from these resistant tumors led to decreasing sensitivity to chemotherapy despite established immunity on the part of the host. The selection of tumor cells resistant to immune destruction rather than drug resistance per se appeared to pay a role. Melphalan was thus able to affect both favorably and adversely the immune factors important to drug-induced regression.


Agglutination reactions of spontaneous canine tumour cells, induced by concanavalin A, demonstrated by an isotopic assay. Quantitative assessment of the agglutination of 51Cr labelled canine cell suspensions to canine kidney cell monolayers has been performed over a range of concanavalin A concentrations. Agglutination was observed with all cell cultures tested, comprising four spontaneous canine melanomas, two canine mammary carcinomas, a benign mammary tumour and a contact-inhibited kidney cell line. The melanomas tested showed strong specific inhibition of concanavalin A agglutination by 10(-2)M alpha-methyl-D-glucopyranoside. Inhibition of agglutination of mammary tumour and kidney cells was weaker and less specific. Agglutination was inhibited at 4degrees C. Reduced agglutination to glutaraldehyde-fixed mono-layers was observed in the case of mammary tumours but was absent when contact-inhibited kidney cells were tested. The specificity of the reaction for transformed cells and the parameters involved are discussed.


An experimental model for meningeal leukemia in rats (L5222). Effect of treatment with BCNU and cyclophosphamide. An experimental model of meningeal leukemia in rats is developed by intracerebral (IC) inoculation of leukemic cells from the transplantable acute leukemia L5222. The L5222 proliferates exponentially in the central nervous system (CNS) and the disease becomes systemic 2 days following IC inoculation. Chemotherapeutic studies with BCNU and cyclophosphamide yielded cures in a high percentage of cases when treatment began at an early stage of meningeal leukemia. When treatment was started at the advanced stage, only BCNU showed a large number of cures. However, cyclophosphamide resulted in a marked increase of life-span. The activity of cyclophosphamide against meningeal leukemia, which is in contrast to the results obtained by Skipper et. al. (1961) in the L1210 mouse leukemia, suggests that cyclophosphamide crosses in part the blood--brain barrier in a rat bearing meningeal leukemia. After subcutaneous inoculation, BCNU and cyclophosphamide showed the same rate of cures.


Histologic specificity of the effect of age at birth of first child on breast cancer risk. Slides of 531 breast cancers from Glamorgan, Wales, were classified according to the presence or absence of each of three histologic characteristics: stromal infiltration by linear strands of tumor cells, areas of intraductal carcinoma, and areas of lobular carcinoma in situ. The proportion of tumors with each of these characteristics was positively associated with age at first child-birth. Increasing age at first birth had a strong positive effect on the incidence risk of tumors with any of these features, but only a small effect on risk of tumors with none. For parous women, the rate of breast cancer with or without linear strands, with or without areas of intraductal cancer, and without areas of lobular carcinoma in situ was lower than for nulliparous women. Parity appeared not to reduce the risk of tumors with areas of lobular carcinoma in situ.


Breast cancer and use of rauwolfia and other antihypertensive agents in hypertensive patients: a nationwide case-control study in Finland. Two nationwide registers, the Finnish Cancer Registry and a register of persons entitled to free drugs for hypertension, were linked in a case-control study of the association of breast cancer and use of rauwolfia. Cases were all hypertensive patients in whom breast cancer was diagnosed in 1973. To test the association specifically with rauwolfia, controls were hypertensive women matched with the cases for age and geographic area and approximately matched for duration of treatment for hypertension. There were 109 case-control pairs. Use of any physician-prescribed drugs during the year prior to diagnosis of breast cancer was ascertained from original prescriptions. In the first set of analyses the patients were classified according to the drug used during most days of the year ("main antihypertensive agent"). In the second set a person qualified as a user of the respective drug regardless of the amount taken. The relative risks in the use of rauwolfia, methyldopa, another synthetic antihypertensive or a diuretic as main antihypertensive agent all ranged between 0.90 and 1.11. The results based on use of a drug in any amount were similar. Next, pairs in which duration of treatment for hypertension was different for cases and controls were excluded. The relative risk associated with use of rauwolfia as main antihypertensive agent then increased from 1.00 to 1.30 and the risk associated with use of any amount of rauwolfia from 1.16 to 2.14. Simultaneously, the relative risk in the use of digitalis was raised from 1.33 to 2.67 and of nitroglycerin from 1.00 to 1.71. Cases also used more types of antihypertensive agents simultaneously than controls. There was no association between rauwolfia-use and breast cancer in analyses limited to pairs in which neither case nor control used digitalis. Thus, there was not a consistent drug-specific association between rauwolfia-use and breast cancer in hypertensive patients. An underlying association of hypertension, heart disease or its treatment (digitalis) and breast cancer may have confounded some of the results of this and earlier studies. In conclusion, it is unlikely that use of rauwolfia increases the risk of breast cancer.


Affinity of estradiol mustard for estrogen receptors and its enzymatic degradation in uterine and breast cancer cytosols. Estradiol mustard (EM) is the 3,17beta-diester of estradiol-17beta (E2) with the nitrogen mustard derivative chlorphenacyl. The ability of EM to bind to cytoplasmic estrogen receptors was tested by inhibition of the binding of 3H-E2 to rat uterine cytosol at 18 degrees C and 30 degrees C. At both temperatures an inhibition curve was observed in the presence of a large excess of drug, suggesting that the latter has a very weak binding affinity (100,000 times lower than E2). Incubation of uterine cytosol with increasing amounts of 3H-E2 in the presence and absence of an excess of EM indicated that the drug interacted with the receptors at the same sites as E2 (competitive inhibition). Preincubation of uterine cytosol at 18 degrees C with EM induced a progressive reduction of 3H-E2 binding capacity. This reduction also occurred, although to a lesser extent, when long-term incubation of the cytosol with EM was performed in the presence of labelled E2 from the start. The process was faster at 18 degrees C than at 4 degrees C and did not occur with EM preincubated in homogenization buffer. Exchange assays by 3H-E2 of uterine receptors preincubated with labelled E2 and excess EM indicated that the drug-induced inhibition of binding capacity was reversible and produced no apparent alteration of the receptors. Furthermore, the rate of exchange was similar to that observed with receptors previously filled with unlabelled E2. In 9 "receptor-positive" cytosols from human breast cancers, time-course study of the binding of 3H-E2 in the presence of excess of EM yielded similar results as those obtained with rat uterine cytosol. These results show that EM has a very low binding affinity for the extrogen receptors and that it is metabolized into one or several compounds of higher binding affinity. They suggest that EM is probably not significantly concentrated by the estrogen target tissues such as mammary cancers. Therefore, the drug is unlikely to be very valuable in the treatment of breast cancer through a specific mechanism involving concentration by the estrogen receptors.


Oncornavirus-like particles in malignant melanoma and control biopsies. Tests for the presence of oncornavirus-like particles in human biopsies were made by the Spiegelman simultaneous assay for 70S RNA and RNA-dependent DNA polymerase and by detection of 600-900S particles, incorporating 3H-uridine, produced by cultured biopsy cells. Thirty-one malignant melanoma biopsies from 29 patients were studied. Using the simultaneous assay, evidence of virus-like particles was found in 15/26 (58%) of melanoma biopsies, 0/3 naevi pools, 1/4 samples of skin adjacent to melanoma, 0/3 samples of normal adult skin and 0/3 prepuces. The velocity sedimentation technique was shown to be a useful screening test for oncornaviruses in studies of two virus-producing mouse cell lines (TKL-5 and WEHI-22), and was positive with 7/9 melanoma biopsies. Overall, these results are compatible with the earlier findings of similar virus-like particles in malignant melanoma cell lines, but the exact nature of the particles remains to be defined.


Effect of Friend leukemia virus on megakaryocytes and platelets in mice. The thrombocytopenia that follows infection of C3H/Bi mice with Friend leukemia virus (FV) has been investigated. At various times after infection, megakaryocytes in bone marrow and spleen were collected on Nuclepore filters, stained with the Feulgen procedure, and counted. At the same times, platelets in circulating blood were enumerated by the direct method of Brecher et al., 1963 We found that megakaryocyte numbers decreased within 3 days of infection; platelet numbers began to fall at 7 to 9 days. At 11 days megakaryocyte numbers in femoral marrow and spleen were reduced to less than one-third of control values, platelets to about one-quarter of control values. The data are compatible with the hypothesis that the thrombocytopenia which occurs after FV infection in mice results from the reduction in megakaryocytes that is brought about by the infection. Whether all classes of megakaryocytes are equally affected by FV was next examined. At various times following infection, megakaryocytes in marrow suspensions were concentrated by unit gravity sedimentation and microspectrophotometric measurements of nuclear DNA content were made. The kinetic data showed that, following FV infection, megakaryocytes of high nuclear DNA content were markedly reduced in number, but those of low nuclear DNA content were relatively unaffected. Thus the virus infection may interfere with the normal sequential doubling in DNA content of megakaryocytes and/or it may lead to preferential elimination by the host of those megakaryocytes in the higher DNA classes.


Tumor antigen extracts stimulate cytotoxicity against unrelated target cells. A new cytotoxicity assay for the study of tumor antigens is described. Splenocytes from normal and mammary-tumor-bearing BALB/c mice were depleted of glass-adherent cells, exposed to various concentrations of tumor extracts and mixed with 51Cr-labelled chick red blood cells. Cytolysis evidenced by 51Cr release was determined 20 h after incubation at 37 degrees C. Lymphocytes from tumor bearers exerted no cytotoxic action per se, but became cytolytic after confrontation with immunogenic tumor extracts. Although this cytotoxicity is expressed against targets other than the cells containing the sensitizing antigen, i.e., a non-specific activity, its induction is strictly specific as evidence by the following: (1) extracts of D1-DMBA-3 tumors exerted an inductive effect only in lymphocytes from mice bearing this tumor and not in lymphocytes from normal mice; and (2) extracts of normal tissues or of a non-immunogenic mammary tumor, D1-DMBA-2, did not induce cytotoxicity in lymphocytes from normal or tumor-bearing mice. This approach may provide a useful method for in vitro determination of cell-mediated immunity in which specific reactions can be induced by tumor extracts as opposed to intact tumor cells.


Kinetics of the response of spleen cells from tumor-bearing animals in an in vivo tumor neutralization assay. The behavior of spleen cells from tumor-bearing mice, vis-à-vis isologous tumor cells, was investigated by means of an in vivo adoptive neutralization test. C3H/eB mice were challenged with tumor cells from a chemically induced fibrosarcoma. Spleens from these animals were removed at weekly intervals following tumor inoculation, mixed with tumor cells, and tested for their influence on tumor growth in syngeneic recipient mice. Two phases in the reactivity of spleen cells from tumor-bearing mice were clearly distinguishable. In a first stage of tumor growth, these mice yielded specific tumor-inhibitory cells conferring protection. Subsequently, the protective activity declined leading to a second phase characterized by tumor enhancement. Both protective and enhancing activities were shown to be mainly dependent on the presence of T cells.


Leukaemias in BN/a and BN/b mice after prolonged treatment with antilymphocyte globulin. The influence of prolonged treatment with antilymphocyte globulin on tumour incidence in BN/a and BN/b mice was studied. One ALG sample appeared to be highly leukaemogenic in two independent experiments. All treated mice developed, after a short latency period, non-thymic lymphomas with involvement of abdominal lymph nodes and spleen resulting in ascites. Three established ascites leukaemia cell lines were examined for the presence of MuLV. In all lines viral C-type particles, soluble viral gs-1 antigen and cell surface antigen related to Gross virus were demonstrated. The mechanism of induction of these tumours remains obscure. Thy hypothetical role of immunosuppression, stimulation of lymphoid cells and activation of leukaemogenic virus is discussed.


In vitro sensitization to embryonic antigens. Lymphoid cells from virgin, untreated BALB/c mice cultured 5 days on 12- or 13-day syngeneic mouse embryo fibroblasts (MEF) or on cells from a syngeneic 3-methylcholanthrene-induced mouse sarcoma (1315) were cytotoxic for 1315 tumor target cells when tested with a microcytotoxicity assay. They did not kill embryo or skin fibroblast target cells. Lymphoid cells cultured with 11-, 14- or 17-day MEF were not cytotoxic for cells from the 1315 tumor, from embryo or from adult skin. Lymphoid cells from multiparous BALB/c mice cultured on 11-, 12- or 13-day MEF, on line 1315 sarcoma cells or on skin fibroblasts were cytotoxic for tumor target cells. MEF were killed by multipara lymphoid cells that had been cultured on 11-, 12- or 13-day MEF or on the 1315 tumor line. Multipara lymphoid cells cultured on 1315 cells or 12-day MEF were cytotoxic for adult skin fibroblasts, while multipara lymphoid cells cultured on 14- or 15-day MEF were not cytotoxic for any of the target cells. The data thus indicate that lymphoid cells can mediate both primary and secondary immune reaction in vitro to antigens shared by neoplastic and normal embryonic cells, and that the primary reactions appear to be more specific to putatively embryonic tumor antigen(s) than the secondary ones.


Chemical-viral co-carcinogenesis: requirement for leukemia virus expression in accelerated transformation. The essential role of Rauscher leukemia virus (RLV) multiplication in viral-chemical co-carcinogenesis was investigated by the use of ethidium bromide (EtBr) as an inhibitor of viral complementary DNA (cDNA) integration in the host genome. EtBr inhibited co-carcinogenic transformation when present at the time of RLV inoculation but was ineffective when added to preinfected cells. Inhibitors of protein synthesis, puromycin and cyclohexamide also inhibited co-carcinogenic transformation of chronically infected cells. Purified rat interferon used at a concentration which inhibited 85% of RLV production did not modify the course of co-carcinogenic transformation. The implications of these observations in terms of the possible role of the virus-specific protein (s) in the co-carcinogenic process are discussed.


Comprehensive alcohol treatment planning, implementation, and evaluation. A descriptive overview of a developing comprehensive alcohol treatment program is presented. The influence of staffing practices and client recruitment on implementation of treatment components is reviewed. Program evaluation and current issues regarding such evaluation are discussed.


The imputation of motives for marijuana behavior. The motives imputed to marijuana use and nonuse by both users and nonusers were investigated by means of open-ended questions. Verbal responses were categorized to symbolize typal vocabularies of motive, and these categories were tabulated against behavioral, social, experiential, and attitudinal variables. The results indicated that frequency of marijuana use and favorableness in drug attitude are the key factors that influence the degree to which different vocabular explanations are offered. Current users most often say that personal experience and personal reward are the motives behind use of the drug, and that legal sanctions and psychomedical effects largely account for its nonuse. Nonusers with unfavorable attitudes tend to verbalize emotional relief and psychological dependence as the motivations for use, and emotional independence for nonuse. Some of the vocabularies of both of these groups are emphasized by former users and nonusers with favorable attitudes. Interpretations of the data in terms of sociological theories of motives are presented.


Methaqualone misuse: foreign experience and United States drug control policy. In 1972 methaqualone emerged as a major drug of nonmedical use in the United States--a subject of widespread publicity and public concern. In late 1973, government officials responded by taking the unprecedented measure of imposing the strictest controls available under United States law on a drug which had previously been subject only to a simple prescription requirement. Methaqualone had a similar history in other countries, particularly in Germany, Japan, and Great Britain. However, this history was ignored by United States officials until nonmedical methaqualone use had become a substantial problem in the United States.


Ethnosemantic analysis of cognition in the alcoholic patient. Ethnosemantic analysis, a research method developed by structural anthropologists and linguists for studying "alien" cognitive domains, would be a useful tool for therapists who treat the alcoholic patient. In this paper a method is developed that would enable a therapist to focus upon aspects of the alcoholic patient's cognitions that are operative in the patient's drinking behavior. Three sorts of objectively derived information are obtained: (1) a list of cognitive factors that threaten the patient's self-concept, (2) a taxonomic sorting of these factors, and (3) a measure of the differential stressfulness of these factors to the patient. All three of these information outputs are semantically produced by the patients themselves.


21-day outpatient methadone detoxification: an evaluation. A population consisting of all consecutive admissions between May 29, 1974 and July 30, 1974 was studied. Of the 169 applicants, 63 began the detoxification regimen. Persons participating in this program were in contact with the program only during their 21-day detoxification episode. The treatment setting was highly permissive. The primary objective was to help and assist individuals in their efforts to either reduce their use of opiates or attain a drug-free state. Thirty-one of the 63 individuals completed the program. Forty-five persons had lower opiate readings when their last three urine samples were analyzed. Interpretation of the data is guarded.


Dimensions of change in the improving alcoholic. An inventory encompassing six social-psychological dimensions that have been found to be related to problem drinking (Attitude toward Drinking, Environmental Support for Heavy Drinking, Impulsivity and Nonconformity, Alienation and Maladjustment, Looseness of Social Controls, and Unfavorable Expectations) was used to determine what areas of behavior were seen to change as an alcoholic improved. Three groups of social workers, psychologists, and psychiatrists rated an alcoholic just hospitalized for treatment, an alcoholic just released from a hospital treatment program as improved, and an alcoholic who had been hospitalized for treatment, had improved, and had remained sober for 4 months. These professionals concurred that change occurred only on Attitude toward Drinking.


Recidivism in youthful heroin offenders and characteristics of parole behavior and environment. An investigation was made of the association between the postrelease circumstances and behavior of 79 youthful heroin offenders and parole outcome. The data suggest that steady employment and the absence of drug use on parole are significantly related to parole success and to each other.


From heroin to methadone--social role changes and reinforcement differentials in relation to outcome on methadone. Part I. The study of reinforcement differentials. This study examines the relevance of reinforcements available to the addict in five critical stages of his career to his success on methadone maintenance. In-depth focused interviews were held with 30 addicts who had completed a program of methadone induction and who were characterized by social worker's evaluations as Successful, Marginally Adjusted, or Failures. The Successful group showed a higher initial negative reinforcement followed by higher positive reinforcement on methadone. The Moderately Successful group showed more negative reinforcement before treatment but continued to show negative reinforcements afterwards. The Failures were low on negative reinforcements earlier but negative reinforcements were high after treatment.


Employing ex-addicts: determinants of support and opposition. In 1973 public hearings of the Temporary (N.Y.) State Commission to Evaluate the Drug Laws concluded with a report which documented pervasive controversy and discrimination in employing rehabilitated or ex-addicts. To analyze local attitudes to this issue, the present study targeted the audience of a recent Symposium "Employment of the Ex-Addict", held in Westchester County, New York, October, 1973. Ss (N = 83) completed a questionnaire to assess the nature and extent of proponent views, degree of commitment, and motivational components of support and opposition. The data strongly supported belief in the urgency of the need to employ. Educational and community measures of moderate activism were preferred to antidiscrimination legislation. Significant correlations were found between urgency and age, size of plant, and occupational group. Appeals to human rights and social rights were judged to be important positive reinforcers to employment, and the economic argument a greater deterrent than fear or prejudice. Study limitations, findings, and need for further behavioral organizational research were discussed.


Lichen planus in India: an appraisal of 441 cases. An analytical study of 441 patients with lichen planus is presented. Lichen planus occurrence was 76% of conditions seen at this institution. The sex ratio was 3:2. Most patients having the disease were in the 3rd decade of life. Lichen planus vulgaris (common type) was seen in 329 (74%) patients, lichen planus hypertrophicus in 56 (13%), lichen planus actinicus in 33 (7.5%), lichen planus follicularis in 8 (2%), lichen planus atrophicus in 8 (2%) and lichen planus pemphigoides in 7 (1.5%) patients. Mucous membrane involvement was seen in 127 (29%) cases and nail involvement in only 7 (1.5%).


Chronic zinc deficiency syndrome in a beer drinker with a Billroth II resection. A 51-year-old woman, addicted to beer for several years, developed chronic zinc deficiency which caused severe mental impairment, a poor general condition with edema and diarrhea, widespread eczema craquelé and loss of hair. Laboratory tests showed a significantly lowered serum zinc concentration, microcytic anemia, an inversed serum albumin/gamma-globulin ratio and a decreased serum thyroxine. Two years earlier, her external pancreatic function had been found severely impaired. Following oral therapy with zinc sulfate 0.2 g X 3R, daily, the patient's mental and physical condition totally changed. In two weeks serum zinc was within the normal range, while hemoglobin concentration, serum proteins, serum thyroxine became normal in 4-8 weeks.


Treatment of zoster and postzoster neuralgia by the intralesional injection of triamcinolone: a computer analysis of 199 cases. On the basis of this study of 111 patients with herpes zoster and 88 with postherpetic neuralgia, it is suggested that the intradermal injection of triamcinolone in saline is a valuable treatment. Mild complications were pain, hemorrhage, abscesses, atrophy, moon face and possibly thrombophlebitis. Zoster patients required treatment for about half as long as those in previously reported control series. In patients treated for active herpes zoster, postzoster neuralgia occurred with about one-third of the frequency noted in other series. In postzoster neuralgia, the patient was benefited sufficiently in 62.5% of the cases to find that life was worth living again.


Age-specific incidence of cancer of the endometrium, ovary and breast in the United Kingdom and the United States. The published data for overall cancer incidence for the registries of Birmingham (UK) and Connecticut (US) show remarkable similarity for men but diverge for women. The incidence of cancer of the endometrium, ovary and breast in Connecticut is higher than in Birmingham, and in each case the menopausal dislocation in the age-specific incidence plot of the Birmingham data is obscured in that for Connecticut. For endometrial cancer, the difference correlates with differences in the two countries in the use of oestrogen replacement therapy, recently implicated in the aetiology of endometrial cancer. The similarity in the pattern for ovarian and breast cancer, and the changing pattern of breast cancer incidence in Birmingham suggest a similar aetiological effect.


Problems and new directions in the evaluation of primary care. The nature of many contacts between patients and first-contact doctors (e.g. general practitioners) is such that the traditional morbidity and mortality indices of outcome are inadequate for evaluating the care provided. Preliminary results of a study developing a more appropriate index of the outcome of primary care, expressed in terms of the patient's functional abilities, are discussed. Beyond its use in evaluating the social and psychological components of care, the method may be applicable in population health surveys and in estimating the need for care.


Validity of questionnaire criteria in mass screening for the diagnosis of peptic ulcer. Epidemiological studies of peptic ulceration in the stomach and duodenum based on a standard questionnaire were carried out among the employees of a sulphur mine. The results obtained in the preliminary cross-sectional study were checked against a sample of 180 people who had undergone radiological examination of the alimentary tract. The validity of the questionnaire was then assessed by comparison of these two sets of results. The study shows that a more revealing screening test may be based on a wide range of questions rather than on a smaller set of specific questions.


A systems approach to cardiovascular disease research. Research in cardiovascular disease is fragmented and difficulties arise in communication between workers in different disciplines. There is no shortage of facts but rather a deficiency in concepts which can accommodate the facts. Systems analysis is suggested as a way to develop a conceptual framework for the assimilation of existing and new information, and to give a basis for decisions about priorities in research. A model of mechanisms in cardiovascular disease is presented to illustrate the approach.


Ringworm infection among agricultural workers. This paper describes a study of the occurrence of T. mentagrophytes var. gran (ringworm) infection in small mammals living in nature and the possibility of the transmission of the aetiological agent from the natural focus of infection to man. The epizootological importance of small mammals is considered from the point of view of their ecological valency, an essential factor in the process of inter-species spread of infection. The concentration of small mammals during winter months in the proximity of human dwellings is due to their search for food and shelter. The barns--the main work place of agricultural workers in winter--were found to contain the highest concentration of small mammals, of the most diverse species and biotopes. They are considered to be the main link in the process of transmission of T. mentagrophytes infection from natural foci to man.


On the catalytic activity of chemically modified enzymes involving two or more substrates and products. Catalytic activity of both native and chemically modified enzymes are numerically expressed in the literature with either kmax (= V/[ET]), or ke (=(V/[ET])/Km), if they refer to a one-substrate one-product reaction. Higher-order catalytic constants of the k'E type have not so far been calculated for enzyme reactions involving two or more substrates and products. a) It is shown here how such higher-order k'E-values can be calculated from easily obtainable experimental information; whatever the number of substrates and products, and even if binding of substrates and release of products follow a branched pathway. b) k'E-Values define the specific rate with which free substrates are converted by the enzyme into free products. Their dimensions (M-ns-1) are defined by the number of substrates (n). k'E-Values refer to the same process as the catalytic constants generally used in physical organic chemistry. c) The required experimental information is mainly based on steady state parameters of the V'- and K'm-types, supplemented with equilibrium and fast kinetic studies of straightforward substrate binding reactions. d) It is shown that higher-order k'E-values always involve the "Haldane problem" of the chemical reaction step. On the other hand, k'E-values are independent of the rate of release of all but the first product in the overall reaction. e) The so-called "Haldane relations" are shown to be determined by the way in which k'E-values can be calculated from V' and K'm-type experimental parameters. f) A number of theoretical considerations indicate that catalytic of the k'E-type are much more suitable for expressing the catalytic activity of chemically modified enzymes than maximal velocities are. This conclusion is demonstrated by a comparison of k'E- and k'max-values of some recently described modified forms of alcohol dehydrogenase.


Conformational analysis of a chain reversal in alpha-chymotrypsin. A complete conformational analysis of the fold (Asp-Lys-Thr-Gly) (residues 35-38), and additional adjacent residues of alpha-chymotrypsin has been performed. A comparison of these findings with those of Lewis et al. (1) is made, and a discussion of the implications of protein-fold models is discussed. This particular residue sequence prefers to bend over maintaining a helical conformation. However, the bend conformation of the tetramer is different from that of the native bend. The native bend conformation is nearly realized when an additional residue of the native primary structure is added to each side of the tetramer. Early and late folding-sequence studies suggests that while the native fold is of low energy, there are fold-points along the primary structure which are more stable. The structural implications of this finding are discussed.


Theory of large-amplitude conformational fluctuations in native globular proteins. Independent fluctuating site model. A theory is developed about large-amplitude conformational fluctuations in globular proteins in their native or predenaturational state. A model is introduced, an independent fluctuating site model, in which we assume that there is more than one independent fluctuating site, each one localized in some part of a protein molecule. Without assuming any further details for each fluctuating site, the entropy S versus enthalpy H curve of this model is shown to be convex. From this fact the predenaturational excess heat capacity can be derived, as observed in recent experimental studies of a few systems of a protein in solution.