Spontaneous cell-mediated cytotoxicity (SCMC) associated with lymphocytes negative for acid alpha-naphthyl acetate esterase (ANAE) activity. A modified histochemical procedure for nonspecific acid alpha-naphthyl acetate esterase (ANAE) activity in human lymphocytes was used to identify a subpopulation of E-rosette forming cells. Performing a one hour reaction at pH 6.5 the distinct dot-like staining pattern was almost exclusively observed on high affinity E-rosettes which sedimented readily in a regular Ficoll-Metrizoate gradient. By combining latex phagocytosis with staining for ANAE activity, a clear-cut distinction between mononuclear phagocytes and lymphocytes could be made. An attempt was undertaken to relate the ANAE marker on human lymphocytes to their functional capacity in spontaneous cell-mediated cytotoxicity (SCMC) reactions. Using as targets two allogeneic (K562,IGR3) and a xenogeneic cell line (L1210) it could be clearly demonstrated that high SCMC activity is mediated by ANAE negative mononuclear cells, whereas enrichment for ANAE positive lymphocytes resulted in a loss of SCMC.


A cytologic aid to diagnosing congenital glaucoma. Using the Giemsa and Papanicolaou staining methods, the authors studied the superficial corneal epithelium of 17 glaucoma patients and 25 normal patients. Smears of the normal patients exhibited a high frequency of precornified cells and a paucity of cornified cells with pyknotic nuclei. Smears of the glaucomatous patients illustrated striking tinctorial changes; these changes were so pronounced that examination of a single section strongly suggested a diagnosis of glaucoma. A case report is presented in which a preliminary diagnosis of congenital glaucoma was disproved by these staining techniques.


Role of radiation therapy in rectal cancer. The advantages and disadvantages of radiation therapy as an adjuvant to surgical resection in potentially curable rectal cancers is summarized by the authors, who are participating in clinical trials to evaluate results.


[Use of electromyography of the bladder sphincter for the differentiation of prostate adenoma and prostate carcinoma]. 82% of the male patients with healthy urogenital tract showed an isoelectric line in the simultaneous EMG-derivation of the sphincter of the bladder under conditions of rest; in 18% of the males a moderate activity was present. In voluntary innervation at once an intensive electric activity of a spindle-shaped discontinuous pattern of the action potential developed in 76% of the patients. In the pressing trial in 83% of the cases a slight to moderately strong activity appeared. In contrast to the examined patients with healthy outlet of the vesicle the patients in whom an adenoma of the prostate gland or a carcinoma of the prostate gland, respectively, was present exhibited deviations from the normal pattern of the action potential, in which cases carriers of adenoma and carcinoma differed. Whether and to what extent the electromyography on the m. sphincter vesicae allows the possibility of the differentiation between adenoma of the prostate gland and carcinoma of the prostate gland in the early clinical stage shall be reserved for further examination.


[Selective demonstration of amyloid deposits and methodical possibilities of analysis of their ultrastructural differences (author's transl)]. In preparations stained with congo-rot and covered with arabic gum amyloid deposits reveal intensive, positive birefringence. Collagen is isotropic or shows a mild negative birefringence. This method is most suitable for selective optical demonstration of amyloid deposits. On the base of investigations on ultrastructural stability of amyloid deposits (resistance to performic acid and induced proteolysis) two types may be distinguished: (1) amyloid deposits of labile structure (generalized, secondary amyloidosis) and (2) amyloid deposits of stable structure (generalized primary amyloidosis, local and senile amyloid deposits).


[Effectiveness of supplemental bleomycin therapy in vulvar cancer]. It is reported about the application of  bleomycin within a randomized prospective investigation. Bleomycin was used for additional chemo-therapy at the vulva-carcinoma. There were treated with Bleomycin 27 of 55 women from two gynaecological hospitals. -- After a period of two years and a half there can not be shown by the "life-table-method" advantage neither in comparison to the only surgical therapy not to the cryo-surgical treatment.


[Status and developmental trends of immunologic contraceptive technics]. The development of contraceptive vaccines is matter of international research. Among antigens possible unique to the reproductive process, sperm antigens, antigens of seminal plasma and of the ovum, antigens of placental hormones and of placenta specific proteins appear suitable targets of the production of fertility regulating vaccines. High specifity of these vaccines can prevent cross reaction with other body cells.


[Serum concentration of the pregnancy-associated alpha2 glycoprotein (alpha2-PAG, protein of the "pregnancy zone", PZ) in malignant gynecologic diseases]. The concentration of pregnancy-associated alpha2-glycoprotein (alpha2-PAG, PZ) was investigated by means of electroimmunodiffusion and single radial immunodiffusion in sera of 30 patients suffering from malignant genital diseases. Results of electroimmunodiffusion indicate that surgical and/or radiological treatment is followed by a significant decrease of PZ serum level. But few months later values have been measured as detected before therapy. There were no significant differences of PZ levels in dependence on localization, histological findings and stage of gynaecological cancer. These findings indicate that periodic determinations of serum PZ do not supply the diagnosis for controlling the course of gynaecological cancer.


[Improvement in the primary immunization scheme for children against whooping cough, diphtheria and tetanus]. The authors studied the reactigenic properties, immunological and epidemiological efficacy of vaccination of children aged under one year by the experimental schemes (AKDC-AKDC-K -- 1 ml, and AKDC-AKDC-K -- 0.5 ml) in comparison with the official pertussis, diphtheria, and tetanus immunization scheme. There proved to be a greater immunogenicity and a better protective effect against pertussis in vaccination according to the experimental schemes. The vaccine reactions were somewhat more incident in children vaccinated according to the AKDC-AKDC-K -- 1 ml scheme, but their essential reduction was reached with reduction of the pertussis antigen third vaccination dose to 0.5 ml.


Biochemical and histological studies on prostates in castrated dogs after treatment with androstanediol, oestradiol and cyproterone acetate. The effect of cyproterone acetate (CA) on experimentally induced benign prostatic hyperplasia (BPH) in the castrated dog was investigated. BPH was induced by 6 months' treatment with 3 alpha-androstanediol (3 alpha-diol) alone and in combination with 17 beta-oestradiol (Oe2). RNA, DNA and zinc content of the glands were determined in addition to histological examination and measurement of the prostates. Two different types of prostatic enlargement were observed. First, 3 alpha-diol induced typical diffuse canine hyperplasia with replacement of functional activity. DNA, RNA and the zinc content of total glands were increased compared with intact controls. Second, 3 alpha-diol plus Oe2 produced on the one hand a more striking increase of prostatic weights, but on the other a loss of typical morphological structure and function. Histologically, transformation of simple glandular epithelium into stratified squamous metaplasia occurred in addition to stimulation of fibromuscular tissue. Biochemically, a relative decrease of DNA per mg tissue was measured with a fall in the RNA to DNA ratio and zinc to the values of castrates. Administration of CA resulted in an abolition of the 3 alpha-diol effect. Biochemical determinations and histological examinations revealed an effect similar to castration after treatment with 3 alpha-diol plus CA. After treatment with 3 alpha-diol plus Oe2 plus CA fibromuscular stimulation as an oestrogen effect predominated in addition to glandular atrophy and metaplastic changes, especially in prostatic ducts. Epithelial hyperplasia is an effect of 3 alpha-diol, whereas metaplastic proliferation only occurs in oestrogenized and androgenized dogs. In both types of prostatic enlargement CA prevents development of hyperplastic prostate.


Regulatory mechanisms of alpha-fetoprotein production in injured rat liver. The role of alpha-fetoprotein (AFP)-suppressing factor(s) in the increased production of AFP in liver injury was investigated with carbon tetrachloride and ethionine-treated rats. Elevated concentrations of serum AFP induced by these treatments decreased upon administration of prednisolone. However, injections of methionine and adenosine 5'-triphosphate (ATP), which significantly accelerated the rapid fall of serum AFP levels after birth, produced no such effects on increased AFP levels in hepatic injury. This ineffectiveness of methionine and ATP may suggests that their metabolisms are impaired in the injured liver.


Histamine-release in rat gastric mucosa by pentagastrin. The release of histamine from isolated gastric mucosa of the rat was studied by a superfusion model. Pentagastrin increased the basal efflux of histamine in a dose dependent manner. Secretin failed to influence the release of histamine under similar conditions. The results are compatible with the view that in gastric mucosa pentagastrin releases histamine which then may react with the receptors of the parietal cell thus serving as mediator in the acid secretory process.


Uptake of horseradish peroxidase in sensory nerve terminals of mouse trigeminal nerve. Uptake of horseradish peroxidase (HRP) in sensory nerve terminals in sinus hair in the nose of the mice was studied after local (s.c.) or i.v. injections. HRP spread into the sinus hair follicles and surrounded the nerve terminals of the Merkel disc endings and lanceolate terminals. It was incorporated into vacuoles and vesicles of these terminals. Subsequently, HRP was demonstrated in nerve cell bodies in the ophthalmo-maxillary part of the trigeminal ganglion, indicating a somatopetal axonal transport of the tracer. In this way the sensory neurons may be subjected to various influences from the periphery.


Improved Nissl method to stain formaldehyde or glutaraldehyde-fixed material. Nissl staining of paraffin sections from formaldehyde- or glutaraldehyde-fixed specimens is significantly intensified when sections are kept in a 50% (w/v) aqueous solution of potassium metabisulfite before being stained by a conventional Nissl method.


Human acoustic neurinomas: nervous system specific biochemical parameters. A series of 24 human acoustic neurinomas from 24 patients has been assayed for several biochemical parameters characteristic of the nervous system. S 100 protein, 2', 3'-cyclic nucleotide 3'-phosphohydrolase activity, and the myelin lipids galactosylceramide and sulfogalactosylceramide (sulfatide). Myelin basic protein was not detected. These finding further support the neuroectodermal origin of the human acoustic neurinoma, and provide additional biochemical markers for further study.


Glial fibrillary acidic protein and intermediate filaments in human glioma cells. Cultured human glioma cells were studied by double indirect immunofluorescence technique using antisera against intermediate filaments and glial fibrillary acidic protein. With both antisera cytoplasmic fibrillar fluorescence was seen. Perinuclear bundles of intermediate-sized filaments, induced by vinblastine treatment, were strongly stained with both antisera. The degree of codistribution of the two types of antigenic determinants varied considerably from cell to cell. The results suggest that two types of filament-related antigenic determinants can be present in the same cell, and also that glial fibrillary acidic protein-related filaments may possess functional similarities to the intermediate filaments found in other cells. Glial fibrillary acidic protein remains as a useful and specific antigenic marker for the study of glial cells in vitro.


Type C particles in culture of human glioblastoma cells. A long-term cell culture of human glioblastoma was investigated microscopically, virologically, and biochemically. Reverse transcriptase activity was detected in cultured human glioblastoma cells. 3H uridine was incorporated into particles of buoyant density at 1.07 g/ml (Ficoll) which is equal to that of Oncorna virus particles, but 3H thymidine was not incorporated at all. Furthermore, reverse transcriptase activity was also demonstrated with the particles, suggesting that the cultured human glioblastoma cells were producing type C Oncorna virus. Ultrastructural observations of cell culture of glioblastoma showed type C virus particles in cisternae and culture medium. Budding of the virus was also seen on the surface of the cell. The mean diameter of the particles was approximately 100 nm. Ca. 1.1 nm of spikes protruded from the envelope. Both types of virions were observed, i.e. the doughnut-shaped type form and the solid circular form.


Eosinophilic globoid body of the pancreas. Histological, histochemical and ultrastructural studies of the eosinophilic globoid body of the human pancreas are described. It had a central cavity surrounded by a collarband measuring approximately 70 micrometers in diameter and is definitely different from cross section of small arteries. The complex ultrastructure was revealed with electron microscope in the eosinophilic globoid body. This consisted of various patterns of unit membranes. Some of the bodies are probably derived from the membrane systems of the pancreatic acinar cells.


New aspects of the morphology of circulating megakaryocytes in rabbits. Circulating cytoplasmatic megakaryocytes, isolated from central venous blood of rabbits by a saponinhaemolysis leuco-concentration technique, were found to have two morphological appearances: a globular and an oblong form, referring to the shape of the cells as a whole as well as to the shape of the nuclei. The nucleus was either centrally or peripherally located. Of the megakaryocytes observed, 40% were cytoplasmatic and 60% naked nuclei. Regressive changes in the residual nucleus was observed, only 1.5% showing karyorrhexis or karyolysis. Megakaryoblasts and promegakaryocytes were never encountered. The average megakarycyte content per ml central venous blood was 3.8 (range: 0-10). A not previously described cytoplasmatic-ligating process, in which the nucleus seemed to move out of the cytoplasm, forming a naked nucleus and leaving the total cytoplasm intact, was observed. We call this phenomenon "nucleus escape". The function and fate of the intravascular megakaryocytes are briefly discussed.


Infantile vaginal tumour with alpha-fetoprotein synghesis. A 17-month-old girl with a vaginal embryonal carcinoma (germ cell tumour) is reported. Alpha-fetoprtoein (AFP) was elevated before treatment in blood and vaginal fluid. Normal levels of AFP in blood and vaginal fluid were found after radiation and comination chemotherapy. By an indirect immunoperoxidase staining AFP was demonstrated in tumour tissue. 24-month-old local recurrence was preceded by a slight increase in serum AFP, and AFP could also be demonstrated in vaginal fluid and vaginal smears.


Suppressor cells for in vivo cytotoxic responses--regulation of the in vivo activation of cytotoxic T-lymphocytes by suppressive cells. A significant in vivo activation of cytotoxic T-lymphocytes (CTL) against trinitrophenyl (TNP)-modified autologous cells and of a DNA-synthesis response in the peripheral lymphnodes is observed in cyclophosphamide (CyP) treated mice after skinpainting with trinitrochlorbenzene (TNCB) or after injection of TNP-coupled spleen cells (TNP-Spl) into the footpads. The activation of these responses can be suppressed by the transfer of spleen cells or lymphnode cells from skinpainted normal mice, but not from skinpainted mice that had been pretreated with CyP. Suppressive activity is also induced by injections of TNP-Spl i.p. or trinitrobenzosulfonate (TNBS) i.v. Optimal activation of suppression occurs with 3--4 days. The suppressive activity is antigen-specific at least in respect to its activation. Suppressor cells of this kind also suppress the induction of delayed hypersensitivity (DH) responses and the priming for in vitro secondary responses. However, these two responses are less sensitive to the suppression, and their in vivo activation is accordingly much less restricted with the in vivo activation of DNA-synthesis and primary CTL responses. DH and CMC memory can be activated ty TNCB skinpainting without pretreatment with CyP.


A tumor T cell line which can be specifically activated by antigen. We have shown that permanently transformed T cells of a leukemic line (485-2) maintain their immunocompetent function and can be specifically activated by antigen. This cell line should facilitate investigations into the biochemical mechanisms of T cell activation and may help elucidate the nature of the T cell receptor.


On the mechanism of histamine release induced by thapsigargin from Thapsia garganica L. Thapsigargin (Tg) is a pure chemical compound isolated from Thapsia garganica with a molecular weight of 650. It releases histamine from isolated rat mast cells but not from isolated histamine-retaining mast cell granules. The rate of release is markedly influenced by pretreatment of mast cells with Tg prior to the addition of calcium. In agreement with the effect of the ionophore A23187 but in contrast to many other calcium-dependent histamine-releasing agents, cells preincubated with Tg respond to the secretory action of calcium whenever the ion is introduced. However, after dilution of Tg-pretreated cells histamine release induced by the addition of calcium became dependent on the time of its addition. The secretory reaction induced by Tg and calcium can be divided into a two-step reaction at 37 degrees C. Pretreatment of mass cells with Tg renders the cells insensitive to the secretory action of compound 48/80 in the absence of calcium, and this effect could be partly counteracted if 1 mM of strontium was added together with compound 48/80. It is concluded that among various calcium- and energy-dependent histamine-releasing agents Tg most closely resembles the action of fluoride on isolated rat mast cells.


Diagnostic dilemma of a 13-month-old boy with 'late-onset' combined immunodeficiency. A 13-month-old boy had a "late-onset" form of combined immunodeficiency and a fulminant Pneumocystis carinii pneumonia of one month's duration. There was no evidence of cutaneous-delayed hypersensitivity responses to diphtheria-tetanus toxoids, Candida albicans, or streptokinase-streptodornase, or of lymphocyte DNA synthesis after in vitro stimulation with the mitogens phytohemagglutinin and concanavalin A, and only 2% to 4% of peripheral blood E-rosetted T lymphocytes. The serum IgM level was normal (62 mg/dL), whereas the other immunoglobulins were markedly reduced. Despite an increased number of Ig-bearing lymphocytes, in vitro Ig secretion after pokeweed mitogen stimulation was substantially reduced. The thymus gland was dysplastic with no Hassalls' corpuscles or thymocytes, and other lymphoid organs showed depletion of T-dependent areas to a greater extent than the B-dependent areas.


Human fibroblasts produce "pregnancy-specific" beta-1 glycoprotein in vitro. Normal and genetic variant strains of nonmalignant human fibroblasts produce in vitro a substance indistinguishable from the recently described "pregnancy-specific" beta-1 glycoprotein (PSbetaG, PAPP-C, SP1). Demonstration of this presumed placenta-specific material in a nonplacental site raises questions about its normal physiologic role and use as a cancer marker.


Climate and the evolution of skull metrics in man. Two types of data, anthropometric and gene frequencies, may be used to reconstruct human evolution. Previous research, reconstructing the history of racial differentiation on the basis of gene frequencies, indicated that the major separation between human groups was that between Africans and Europeans on one side and peoples from Australia, East Asia and Americas on the other. A similar attempt by Howells ('73b), based on skull measurements in 17 ancient populations, in agreement with earlier anthropometric data, showed the major separation to be between Africans and Australians on one side and Europeans, Asians and Americans on the other. Climate could be a contributing factor to the observed differences in skull and anthropometric measurements. Howells' data showed high correlations with several climatic indicators. Carrying out the phylogenetic analysis after elimination by linear regression of the effects of climate has resolved to a great extent the discrepancy between anthropometric and gene frequency data.


Actions of diethylcarbamazine in experimental anaphylaxis. Diethylcarbamazine citrate (DEC) is an anthelmintic drug which has been used recently, with varying results, in the treatment of bronchial asthma. We have examined the pharmacological activity of this drug in several in vivo and in vitro models of experimental anaphylaxis. DEC at doses of 25 and 50 mg per kg given intraperitoneally significantly reduced the mortality rate in sensitized guinea pigs during protracted shock phase but did not modify the time for the appearance of dyspnoea when antigen (egg albumin) was administered as aerosol. Nor did DEC at therapeutic doses inhibit the degranulation and histamine release from rat isolated peritoneal mast cells induced by phospholipase A, compound 48/80, and antigen respectively. It has been suggested that one possible explanation for the apparent effectiveness of DEC in asthma is its ability to antagonize prostaglandin F2 alpha. Our results using in vivo and in vitro models of bronchoconstriction in guinea pigs do not support this hypothesis to explain the inhibition of protacted shock by DEC.


Accumulation of radioiodine in thiouracil-hyperplastic thyroids of chicks. Kinetics of accumulation of radioiodine was studied in thyroids of chickens before, during, and following ingestion of 0.25% thiouracil (TU). After a latent period of about 5 days, weight of the thyroid gland increased, reaching its maximum (42 mg/100 g body wt) after 21 days of TU ingestion; thyroid weight decreased immediately on withdrawal of TU but tended to plateau at a higher level than that of controls. One-way clearance increased by day 4 of ingestion of TU and reached its peak early during hyperplasia; it very quickly reverted to a control level on withdrawal of TU. Exit-rate constant increased markedly during early hyperplasia and decreased to a level less than normal after withdrawal of TU. Concentration of 127I decreased by a factor of 18 by 2 wk of feeding TU; it increased to practically a normal level by 1 wk after withdrawal of TU.


Immunological relationships of Long Island isolates of Babesia microti. Studies to detect strain differences among two rodent-derived and one human-derived Babesia microti isolates from Long Island were undertaken, using various methods. Superinfection experiments using the homologous and heterologous isolates showed cross-protection. All hamsters were resistant to superinfection challenges of increasing dosages of both the homologous and heterologous isolates. Attempts to infect other laboratory animals with the Long Island isolates of B. microti were successful in intact and splenectomized Sprague-Dawley rats and questionable in Swiss mice. Nylar and CFW mice as well as CFW and Wistar intact and splenectomized rats were refractory to B. microti isolates from Long Island. Indirect fluorescence tests using convalescent sera from six Long Island cases of babesiosis showed no titer differences with tests using the three Long Island antigens as well as the Gray strain antigens. The rise of hamster IgG anti-B. microti antibody was followed by indirect immunofluorescence done at different parasitemia levels. The IgG antibody in hamsters was detected early in the course of infection, rose rapidly concurrent with increasing parasitemia, and became stable at high titers for the duration of the infection. IgG antibody titers were unaffected by homologous superinfection challenges.


[Toxic pathology of the hepatocyte in histiotype culture. II--Effects of an organic chlorine compound: lindane. Effect of a foreign metabolizable substance and toxic effect]. Lindane or HCH is gamma-hexachlorocyclohexane, still used as an agricultural insecticide. Tests were carried out using primary cultures of chicken embryo hepatocytes at the 11th day of incubation, for a fixed period of 40 hours, using different concentrations of the substance. From 0.5 to 10 mumol/l, acid phosphatase fell significantly, without any other consequences being observed. From 25 to 50 mumol/l, there was a tendency to hypertrophy of the mitochondria, to hyperplasia of the granular and smooth endoplasmic reticulum, and of the Golgi system. Growth was unchanged and acid phosphatase decreased. From 75 to 125 mumol/l, these changes were accompanied by signs of cellular damage and there was marked autophagia. Growth was reduced and acid phosphatase greatly decreased. From 150 to 175 mumol/l, the lesions were markedly polymorphous and some indicative of impending cytolysis, in particular spiral tearing of the hyaloplasm. Growth was markedly impaired, whilst there was a paradoxal increase in acid phosphatase. In increasing concentration, L first had the effect of a metabolisable foreign substance, of the "phenobarbital" type, then a definite toxic action. Sensitivity of the individual hepatocytes differed.


Failure of mechanical properties to parallel changes in lung connective tissue composition in bleomycin-induced pulmonary fibrosis in hamsters. Lung volumes and volume pressure (V-P) relationships were measured in anesthetized hamsters 8, 30, 60, and 90 days after induction of interstitial pulmonary fibrosis by intratracheal administration of bleomycin. Subsequently, total collagen, elastin, protein, deoxyribonucleic acid (DNA), and dry weight were determined in the lungs of each animal. The mean volume of air in the lungs at a transpulmonary pressure of 25 cm H2O and mean quasi-static compliance were decreased at 8 and 30 days and had returned toward normal by 60 and 90 days. Dry lung weight and total protein content were increased at 8 days, peaked at 30 days, and were still greater than normal at 90 days; DNA peaked at 8 days, remained unchanged through day 60, and returned to normal by day 90. Collagen and elastin content, although not significantly different from control at day 8, was increased at day 30 with peak values attained at day 90. Ratios of collagen or elastin to dry weight, total protein, and DNA were decreased at 8 days, normal at 30 days, and increased at 90 days. The ratios of collagen or elastin to total protein, dry lung weight, or DNA cannot be used as indicators of the amounts of these proteins in the whole lung. We conclude that in interstitial pulmonary fibrosis induced with bleomycin the pattern of changing biochemical composition of the lungs cannot be inferred from the lung volumes or V-P relations.


The doctor's white coat. The image of the physician in modern America. The social perception of the physician has changed during the past century. This is reflected in the changing use of a symbol of the profession, the white coat. This dress originated in operating rooms, scientific laboratories, and modern hospitals, each of which contributed to the social understanding of what it means to be a healer. Adding the cultural significance of "whiteness" reveals a broad spectrum of meaning surrounding the healing encounter whose most important aspects are the authority and supernatural powers of scientific physicians and the protection of patients. A major force aiding in the adoption of this symbol was the shift in sick care from home to hospital, where it served to legitimate otherwise socially taboo behavior. Recent changes in how scientific medicine is publicly viewed have eroded these established meanings. The resulting tensions affect individual patient-physician encounters and interactions between the profession and society as a whole.


[Paroxystic vasomotor skin manifestations (author's transl)]. Paroxystic vasomotor skin manifestations are provoked by various etiologies. Widespread or generalized vasomotor skin manifestations may be induced by a physiological reaction (emotinal flushing), by a drug (vasodilator drugs, antabuse, antidiabetic, sulfonamides), by a discharge of histamine (urticaria, mastocytosis) or by an hypersecretion of serotonin (dumping-syndrome, carcinoid syndrome). They may be caused by an endocrinopathy (menopause, hyperthyroidism, hypoglycaemia, medullary thyroid carcinoma, pheochromocytoma, endocrine pancreas, carcinoma). More rarely vasomotor troubles happen in homocystinuria, inhalation of a toxic (trichlorethylen, calcic cyanamid) and exceptionally in some immunohaematologic diseases. Main localized vasomotor skin manifestations observed are dermographism, facial flushing (Sluder's syndrome, cluster headaches, Frey's syndrome, Riley-Day's syndrome) and acral syndromes (Raynaud's phenomenon, erythromelalgia).


Systemic hyalinosis (juvenile hyaline fibromatosis). Ultrastructure of the hyaline with particular reference to the cross-banded structure. Systematic hyalinosis (juvenile hyaline fibromatosis) is characterized by hyalinized skin lesions. Electron microscopic examination of a hyalinized skin tumor from a 19-year-old man with this syndrome revealed that the hyaline was composed of ruthenium red-positive ultrastructures (granules, filaments, and a kind of cross-banded structure), indicating the presence of glycosamino-glycan or glycoprotein, and a small number of thin collagen fibrils. Using a new ruthenium red staining method combined with an enzymatic digestion procedure, it was demonstrated that the cross-banded structure and granules consisted of chondroitin sulfate-proteoglycan and/or glycoprotein, and that the cross-banded structure is probably a noncollageneous aggregate of the granules attached to the parallel-arranged filaments of hyaluronic acid-like nature.


Mucosubstances in otitis media with effusion. Histochemical localization of sialomucins and sulfated polysaccharides have been demonstrated in the mucous membrane of the middle ear in different middle ear effusions. Inasmuch as a more neutral stain with alcian blue-PAS is still present after treatment with neuraminidase, it is strongly suggested that neutral polysaccharides coexist with neuraminic acid in the middle ear glycoproteins. Sulfated polysaccharides are present only in the mucoid effusions. The use of precise nomenclature for mucosubstances is strongly recommended and is outlined. The nature of the specificity of the histochemical stains is discussed.


Copper stains and the syndrome of primary biliary cirrhosis. Evaluation of staining methods and their usefulness for diagnosis and trials of penicillamine treatment. Use of Shikata's stain was found to be a sensitive method for the demonstration of copper-protein complexes, but use of the rhodamine stain may be more reliable. Interobserver agreement was slightly better with Shikata's stain. Hepatic copper increased with the histologic progression of chronic nonsuppurative destructive cholangitis (CNDC). All specimens that contained Mallory bodies and most specimens (91%) that contained bile yielded positive copper stains. No correlation was found between positive copper stains and other histologic features. A strongly positive copper stain aided in the diagnosis of CNDC. All specimens with negative copper stains contained less than 250 microgram of copper per gram of dry weight. From a strongly positive copper stain, the chemical copper content could not be predicted with certainty. Shikata's method seemed adequate to evaluate penicillamine treatment trials.


The regulative role of food consumption in the induction of rat liver cell proliferation by drugs and environmental pollutants. alpha-hexachlorocyclohexane (alpha-HCH) was used to stimulate cell proliferation in rat liver. Food consumption at two different stages of the replicative cycle was found to be required for the enhancement of DNA synthesis by this agent: (1) Before or at the time of alpha-HCH administration (G0), (2) 12--15 hours later, i.e. 5--8 hours before initiation of DNA synthesis ("R"-point). Other alpha-HCH mediated changes in the liver i.e. organ enlargement and the induction of drug-metabolising enzymes, remained unaffected by the absence of food. This suggests that the nutritional control is specific for induction of DNA synthesis. It is concluded that food consumption, in addition to specific stimuli of liver growth, is an exogenous regulator of liver cell proliferation.


[Structural and functional reactive changes in the bushy interceptor of the frog urinary bladder upon adequate stimulation]. Morphological and functional changes in the bushy receptors of the frog urinary bladder were studied after a definite cystectasia (bladder wall distention). Morphological and electrophysiological parameters were registered synchronously. Under the effect of distention the total rate of afferent impulse activity was demonstrated to increase with a simultaneous increase of the average size of the receptor end-plates and with changing (growing round) in their form. Increment of the average size of the end-plates overtakes that of the distance between two points chosen on the urinary bladder wall; certain shifts are also observed in time parameters for staining the receptors with methylene blue. A possibility is suggested on interconnections of morphological and electrophysiological shifts.


[Obtaining pure antibodies with the aid of an immunosorbent based on sephadex]. A method for production of pure antibodies by means of an immunosorbent based on sephadex according to the technique of Wilson and Nakane modified by the authors is described. The method requires no special equipment, is simple and may be used in any laboratory for immunomorphological studies.


[Tubular aggregates in a case of chronic proximal spinal atrophy]. The case of a 35 years-old man, with chronic proximal muscle atrophy in which at the muscle biopsy tubular aggregates were found by histochemistry procedures is reported. The tubular aggregates stained positive with the modified Gomori trichrome, haematoxylin-eosin, DPNH-diaphorase, non specific esterases, phosphorylase, P.A.S., oil red O and lactate dehydrogenase. They did not show in the routine and acid pre-incubated ATPase, acid and alkaline phosphatases and succinate dehydrogenase. Only found in type II fibers. A brief discussion about the pathogenesis and function of the tubular aggregates is made. The authors believe that the tubular aggregates in this case are secondary to prolonged use of phenobarbital and diphenylhydantoin, associated with the basic denervation process and alcohol abuse.


[Meningeal melanomatosis: cytomorphologic analysis of the cerebrospinal fluid]. Six cases of meningeal melanomatosis are reported. The cytomorphological analysis of cerebrospinal fluid lead to diagnosis in all of them. Emphasis is given to morphological characteristics of the tumor cells and specially to the melanin dark brown pigment, observed in all cases here reported.


The resources centre in the dental school. A brief introduction to the philosophy of resources centre based teaching has been presented. A review of the situation with respect to audio-visual usage in some dental schools in the United Kingdom and in the United States of America has been conducted. It is apparent that the trend towards the use of audio-visual material in dental teaching is increasing in both countries. Concomitant with this increase is the real requirement to develop a management function commensurate with production and usage potential. The implementation of a resources centre based teaching model in the dental school will provide a potential for enormous improvement in teaching efficiency as well as a vast increase in teaching effectiveness in the various dental disciplines. In like manner, the student population will find an opportunity to participate more fully, more positively, and more personally in an enhanced teaching and learning environment.


Search for Epstein-Barr and type C oncornaviruses in systemic lupus erythematosus. Lymphoblastoid cell lines were derived from patients with active systemic lupus erythematosus by allowing spontaneous transformation of peripheral B lymphocytes (B cells) harboring endogenous Epstein-Barr virus or by superinfecting peripheral lymphocytes with exogeneous Epstein-Barr virus. Results of extensive studies aimed at identifying type C oncornaviruses in these lymphoblastoid cells were entirely negative by electron microscopy, DNA-DNA hybridization, reverse transcriptase assays, and cocultivation experiments. These results do not support the postulated association of oncornavirus infection in human systemic erythematosus.


The interaction of short-chain aralkyl alcohols and amines with the erythrocyte membrane. Erythrocytes in isotonic saline are hemolyzed by benzyl alcohol and by 2-phenylethanol, but not by the corresponding amines nor by the ring-or side-chain-hydroxylated analogs. All these compounds could however interact with the erythrocyte membrane since: a) they facilitated the hemolytic effect of benzyl alcohol and/or of phenylelytic effect of benzyl alcohol and/or of phenylethanol; b) they exerted a protective effect against controlled hypotonic hemolysis.


A clinical study of low molecular weight-hydroxyethyl starch, a new plasma expander. 1. The pharmacokinetics of a new plasma expander, low molecular weight-hydroxyethyl starch (LMW-HES) were examined in six normovolaemic men. 2. One hour post-infusion, 13.5% of the total dose of LMW-HES injected was excreted in the urine, 50.2% was present in the intravascular space, and 36.3% was unaccounted for. 3. Twenty-four hours post-infusion, 65.5% of the total dose of injected LMW-HES had been excreted in the urine, 4.1% remained intravascularly, and 30.4% was unaccounted for. 4. The plasma volume increased rapidly from a mean value of 45.7 ml kg-1 to a maximum value of 57.9 ml kg-1 immediately  post-injection, then gradually returned to normal over 24 h. 5. The infusion of an average of 58.1 g had no effect on ESR, renal and hepatic biochemical indices. 6. LMW-HES appears to be safe and effective, and should be of value clinically when rapid and short-lived augmentation of the plasma volume is required.


Brain metastases in malignant teratoma: a review of four years' experience and an assessment of the role of tumour markers. Between 1973 and 1977, 247 patients with malignant teratoma have been treated in two units in London. Seventeen have developed brain metastases, an overall incidence of 6.2%. The median survival from diagnosis of cerebral metastases is 6 weeks and all patients except one have died. The survivor is disease-free 12 months after completing treatment, which included extensive use of chemotherapy, surgery and radiotherapy. Serum gonadotrophin (HCG) and alpha-foetoprotein (AFP) estimations have been performed in 264 patients as a means of monitoring the effects of therapy. In 42 patients (37 of whom had Stage IV disease) the peak HCG level was greater than 10(4) iu/l, and the incidence of brain metastases in this group was 26%, significantly higher than in the group with HCG levels below 10(4) iu/l, for which the incidence of cerebral deposits was 1.8% (P less than 0.0001). No significant correlation was seen between peak AFP levels and the incidence of brain metastasis. With the aim of improving results by earlier diagnosis, cerebrospinal fluid (CSF) specimens have been examined for HCG and AFP levels in 56 subjects, 9 of whom had brain metastases. A serum: CSF HCG ratio less than 40 is an accurate indication of the presence of brain metastases, and may have considerable predictive value. However, false-negative serum: CSF HCG rations (greater than 40) frequently occur in patients with proven brain deposits. Estimation of AFP in spinal fluid has not contributed to the early diagnosis of brain metastases in malignant teratoma.


Spontaneous resolution of lichen myxoedematosus. Lichen myxoedematosus was diagnosed in a 44-year-old woman in 1957. Despite the persistence of an abnormal serum protein, the skin lesions cleared by 1965. The protein abnormality disappeared several years later.


The relation between pregnancy specific beta 1 glycoprotein (SP1) levels and abnormal glucose tolerance in pregnancy. Pregnancy specific beta 1 glycoprotein (SP1) levels were measured in 54 potentially diabetic patients in the third trimester of pregnancy using a laser nephelometric method. All patients were tested by the 50 g oral glucose tolerance test. The results show that a significantly greater number of patients with abnormal glucose tolerance had SP1 levels above the mean compared with patients with normal glucose responses. All patients with a diabetic type response had SP1 levels above the mean. There was no significant difference in SP1 levels in the fasting and two-hour plasma samples. The study suggests that patients with asymptomatic glucose intolerance in pregnancy have higher SP1 levels than patients with normal glucose tolerance. It is possible that one of the factors controlling the placental output of SP1 is the maternal blood glucose level.


Acycloguanosine: antiviral activity in the rabbit cornea. We describe experiments, using the multiple microinoculation technique, to produce superficial herpes simplex keratitis in the rabbit cornea, which showed a potent antiviral effect of acycloguanosine.


Model RNA-directed DNA synthesis by avian myeloblastosis virus DNA polymerase and its associated RNase H. A model RNA template-primer system is described for the study of RNA-directed double-stranded DNA synthesis by purified avian myeloblastosis virus DNA polymerase and its associated RNase H. In the presence of complementary RNA primer, oligo(rI), and the deoxyribonucleoside triphosphates dGTP, dTTP, and dATP, 3'-(rC)30-40-poly(rA) directs the sequential synthesis of poly(dT) and poly(dA) from a specific site at the 3' end of the RNA template. With this model RNA template-primer, optimal conditions for double-stranded DNA synthesis are described. Analysis of the kinetics of DNA synthesis shows that initially there is rapid synthesis of poly(dT). After a brief time lag, poly(dA) synthesis and the DNA polymerase-associated RNase H activity are initiated. While poly(rA) is directing the synthesis of poly(dT), the requirements for DNA synthesis indicate that the newly synthesized poly(dT) is acting as template for poly(dA) synthesis. Furthermore, selective inhibitor studies using NaF show that activation of RNase H is not just a time-related event, but is required for synthesis of the anti-complementary strand of DNA. To determine the specific role of RNase H in this synthetic sequence, the primer for poly(dA) synthesis was investigated. By use of formamide--poly-acrylamide slab gel electrophoresis, it is shown that poly(dT) is not acting as both template and primer for poly(dA) synthesis since no poly(dT)-poly(dA) covalent linkages are observed in radioactive poly(dA) product. Identification of 2',3'-[32P]AMP on paper chromatograms of alkali-treated poly(dA) product synthesized with [alpha-32P]dATP as substrate demonstrates the presence of rAMP-dAMP phosphodiester linkages in the poly(dA) product. Therefore, a new functional role of RNase H is demonstrated in the RNA-directed synthesis of double-stranded DNA. Not only is RNase H responsible for the degradation of poly(rA) following formation of a poly(rA)-poly(dT) hybrid but also the poly(rA)fragments generated are serving as primers for initiation of synthesis of the second strand of the double-stranded DNA.


Enzymic polyadenylation of 5S ribosomal ribonucleic acid and synthesis of a complementary deoxyribonucleic acid. The 5S ribosomal RNA has been isolated, pure and intact, from rat liver (5 mg of 5S RNA from 150g of liver). The 5S RNA serves as a primer for calf thymus poly(A) polymerase with 20% of the efficiency of (Ap)3A. Bacterial 5S RNA and transfer RNA also serve as primers; rat liver 18S and 28S ribosomal RNAs support poly(A) synthesis poorly. Neither the 5S RNA primer nor the appended poly(A) tract is nicked or degraded by poly(A) polymerase, and initiation of poly(A) tracts on 5S RNA primers continues throughout the reaction period. The rate of initiation is dependent on the enzyme concentration; the ATP concentration affects the rate of elongation. The polyadenylated material increases in size over time, with the largest material reaching a size of 6.8 S in 5 h, corresponding to an appended poly(A) tract of 140 nucleotides. Using polyadenylated 5S RNA, oliog(dTY as primer, and avian myeloblastosis virus reverse transcriptase, we synthesized DNA complementary to 5S RNA. The complementary DNA has an apparent molecular weight (in alkaline sucrose gradients) of 4.3 X 10(4). Base composition analysis and nearest-neighbor analysis of the DNA are as expected for a complement of 5S RNA, indicating that the entire 5S sequence is copied. The complementary DNA hybridizes to 5S RNA with a R0t1/2 of 8.9 X 10(-4) mol.s.L-1. No hybrid is formed with Escherichia coli 16S and 23S ribosomal RNA, E. coli 5S ribosomal RNA, yeast transfer RNA, rat liver transfer RNA, or rat liver 18S and 28S RIBOSOMAL RNA. The Tm of the 5S RNA:5S DNA hybrid in 15 mM NaCl containing 1.5 mM sodium citrate is 74 degrees C, 2.5 degrees C below the theoretical melting temperature of a DNA duplex of 60% G + C. Analysis of the hybrid in buoyant density gradients also indicates that hybridization is both specific and precise. The complementary DNA anneals to calf thymus, rat liver, and salmon sperm DNAs but not to E. coli DNA. Annealing of 5S cDNA to calf thymus DNA with a C0t1/2 of 2.1 suggests that there are several thousand 5S RNA genes in the calf thymus genome (haploid). At least that number of 5S RNA genes is present in the salmon sperm genome.


Plasma clearance and liver metabolism of native and asialo-human transcortin in the rat. Plasma kinetics and liver metabolism of iodinated human corticosteroid-binding protein have been studied in ovariectomized female rats. 125I-labeled human corticosteroid-binding globulin prepared by a modified chloramine T reaction was shown to be physically intact and biologically active. Intravenously injected 125I-labeled human corticosteroid-binding globulin was shown to give a complex clearance pattern from the plasma, with half-lives of 7.5 and 51 min. Estrogen injections had no effect on plasma clearance rate. Direct involvement of liver plasma membrane receptors for asialoglycoproteins in 125I-labeled human corticosteroid-binding globulin metabolism was demonstrated in vivo and in vitro using asialofetuin as a competitive inhibitor. 125I labeled human asialo-corticosteroid-binding globulin was cleared from the plasma with a half-life of less than 1 min, while the simultaneous injection of 5 mg asialofetuin maintained the circulating plasma lebels. Asialofetuin also slowed the clearance of intact 125I-labeled human corticosteroid-binding globulin from the plasma (t1/2 = 90 min). Binding of 125I-labeled human asialo-corticosteroid-binding globulin to rat liver plasma membranes in vitro was inhibited in a dose-dependent manner by asialofetuin, but not by intact human corticosteroid-binding globulin or fetuin. 125I-labeled human corticosteroid-binding globulin did not bind significantly to the membranes. It is concluded that human corticosteroid-binding globulin clearance from rat plasma is rapid and that the carbohydrate moiety of human corticosteroid-binding globulin is involved in its clearance and catabolism by the liver.


Ontogenesis of pyruvate kinase in the brain and liver tissues of the rat. The ontogenetic study of pyruvate kinase in the brain and liver tissues was performed in different batches of rats, from the foetus at the 13th day of gestation to the adult subject. -- According to the kinetic study, the shape of the curve is transformed from sigmoid to hyperbolic from the 13th day of foetal life to adulthood in the brain. Hill coefficient increases with the age of animal in the liver tissue. -- According to polyacrylamide gel isoelectrofocusing, a family of four, transitory or definite bands are detected in the foetal brain : they are well defined by their pHi : M4, M3, M2, M1 ; at the adult stage, M1 predominates, M2 is minor. Threee principal bands are distinguished in the liver : two are characteristic of foetal life (Lf and M2), one of adulthood (L). -- According to the immunochemical tests, there are antigenic determinants common to M1, M2, M3 and M4. The confrontation of the first two methods prompts the conclusion that the kinetic of the enzyme (and perhaps its function) varies with the animals age and is linked to its molecular structure. With the third method, it allows to stress the precociousness of the appearance of the common antigenic determinants, simultaneously with immature enzymatic forms. The signification of the kinetic modifications as well as the succession of the isozymes of the M type in a determined order are discussed and the in vivo formation of hydbrids is suggested.


Use of trypsin for rapid and efficient purification of murine sarcoma and leukemia virus. A method is described for purification of MSV-MuLV from culture supernatant of chronically infected 78A,1 rat embryo cell line. This method involves direct polyethylene glycol-NaCl precipitation of the low speed supernatant of culture fluid followed by digestion of the pellet with trypsin. This procedure efficiently disrupts large aggregates which normally entrap most of the virus. Highly purified virus can be obtained in very good yield by a combination of sedimentation velocity and isopycnic centrifugation : yields up to 100 A280 units (17 mg of protein) of purified virus per liter of culture fluid can be observed. This procedure appears well suited for large scale isolation of virion associated enzymatic activities.


[Dynamic model of protein behavior in water. Possible mechanism of association and dissociation of specific complexes]. The proposed mechanism of association and dissociation of specific protein complex with ligands is based on the assumption that thermal fluctuations in the non-polar cavities of protein are interrelated due to the co-operative properties of macromolecules. The non-polar cavities, representing the active centres and clefts in the globule jump from the "open" state to the "closed" one with a certain amount of water being removed to the external medium. It is assumed that the association of ligand with the active centre is accompanied by stabilization of the chest state and destibilization of the open one, dissociation being a reverse process. A rapid change in the state of the active centre under the influence of ligand induces a slow relaxation process resulting in similar alterations in the state of the auxiliary non-polar cavities of protein. As a result, the binding constant of ligand increases, while the free energy of protein decreases. Expressions have been obtained which relate the rate constants of association, dissociation and the binding constant to the rate-constant of the nonpolar cavities transitions from the closed state to the open one, and vice versa. They gave a possibility for a qualitative interpretation of the effect of some specific and non-specific agents upon the stability of immunoglobulins, on the influence of salts upon the association and dissociation of antibody-antigene complexes and on the increase in the constant of binding between antibodies and haptens in the course of prolonged immunisation.


[Compartmentation of gramicidin 5 in membranes of sensitive bacteria and protein-lipid interactions]. Gramicidin S is sorbed on the isolated membranes of granicidin-sensitive Micrococcus lysodeikticus strain. The antibiotic inhibits the membrane malate dehydrogenase within the temperature range of 9--42 degrees C, i.e. under conditions of gel and liquid-crystalline lipid state; however its effect at 10 degrees C is 10 times as low as is observed at 42 degrees C. The inhibitory effect of gramicidin S on malate dehydrogenase can be eliminated and the antibiotic can be removed from the membrane by an excess of different phospholipids. No transfer of the membrane components on exogenous phospholipids is observed. A prolonged (about 2 hrs, 30 degrees C) incubation of the membranes with gramicidin S results in irreversible inactivation of malate dehydrogenase, although the antibiotic can be still eliminated by an addition of phospholipid emulsions. It is suggested that gramicidin S forms complexes with phospholipids, in which the antibiotic is oriented to water. These complexes disturb the lipid-protein interactions, resulting in relaxation of the binding between the boundary phospholipids and proteins, in the loosening of near-protein lipid zones and simultaneous condensation of acid phospholipids in the whole membrane. Destruction of the lipid zone is accompanied by changes in the enzyme activity, by separation of lipid and protein regions and by transphase enzyme transitions (expulsion or immersion). A slow formation of secondary protein-protein associates may be irreversible.


[Heterogeneity in organization of lipids in the bacterial membrane]. The lipids of Micrococcus lysodeikticus membranes were 50%-substituted by phosphatidyl choline using lipid-exchanging proteins isolated from rat liver. The incorporation of phosphatidyl choline into the membrane did not significantly change the malate dehydrogenase activity and the temperature dependence activity in the Arrhenius plots for the enzyme. Gramicidin S--modifier of membrane lipids--had similar effects both on the intact membranes and on the phosphatidyl-enriched membranes. A conclusion is made on structural heterogeneity of the bacterial membrane and on the presence of a boundary lipid fraction, which controls the functioning of malate dehydrogenase and is characterized by a low-rate exchange with other lipids.


[Porton uptake by isolated chloroplasts during cyclic and non-cyclic electron transport catalyzed by photosystem I in the presence of ferredoxin]. Proton uptake by isolated chloroplasts during cyclic electron transport in the presence of ferredoxin and under NADP+ reduction from the ascorbate--TMPD donor pair under anaerobic conditions was studied. It was found that during cyclic transport the proton uptake is less intensive than under NADP+ reduction. In the presence of ATP the proton uptake is increased in the first case and is decreased in the second one. During cyclic transport in the presence of gramicidin D the proton uptake is completely suppressed and under NADP+ reduction is decreased down to 0,08--0,09 mk equiv H+ per mg of chlorophyll, irrespective of ferredoxin or NADP+ concentrations. The role of ferredoxin NADP+ reductase in a proton uptake by thylakoids is discussed.


[Liver structure as a possible factor in the regulation of alpha-fetoprotein synthesis]. AFP-containing hepatocytes were shown to lose the membrane antigen, localized in the region of bile capillaries, due to the mouse liver regeneration induced by CCl4 and paracetamol poisoning. Both in the liver regeneration and early postnatal ontogenesis the cessation of the AFP synthesis in hepatocytes coincides with the appearance of the bile capillary antigen on the cell surface. It is suggested that the AFP synthesis cessation is the result of the arrangement of cell contacts characteristic of the definitive liver balk.


[Immunohistochemical study of the differentiation of the cephalic lobe of the adenohypophysis in chick embryos]. It was shown by means of indirect immunofluorescence that ACTH appeared in the cephalic lobe of the chick embryo pituitary beginning from the 8th day of the development. A new tissue-specific antigen (A3) is revealed, which is localized in the cephalic lobe of adenohypophysis and becomes manifest at the 7th day of embryogenesis. Quantitative analysis of ACTH and A3 localization in the cells of 11-day chick embryo adenohypophyses allows a conclusion that A3 is localized in corticotropic cells.


[Biosynthesis and production of specific beta 1-globulin (SBG) during pregnancy in rats]. It has been shown that pregnancy specific beta 1-globulin (SBG) is found in the rat blood serum on the 5--7th day of pregnancy. SBG reaches the maximum by the end of pregnancy and is not detected already at the 3d--4th day after delivery. The placenta is the site of the SBG synthesis. Other organs of the pregnant rat are not able to incorporate radioactive aminoacids into the protein in vitro.


[Synthesis of alpha-fetoprotein, albumin and transferrin by long-term cultured cells of murine hepatoma]. The capacity of continuous cell of XXIIa mouse hepatoma (strain MHXXIIa) to synthesize alpha-fetoprotein, albumin and transferrin was studied by immunoautoradiography. Albumin and transferrin were detected in the polyethylene glycol concentrated growth medium of hepatoma cells on the 5th year (the 55th month) of their cultivation. alpha-fetoprotein was not found. Only transferrin was revealed in the growth medium of hepa toma cells of the 8th year (the 92d month) of cultivation. Two clonal cultures obtained on the 8th year of hepatoma cell cultivation were also characterized by the ability to synthesize transferrin. The continuous mouse hepatoma cells retained their malignancy. The agar micro-precipitation reaction showed the presence of alpha-fetofetoprotein in lyfogel concentrated serum of mice with tumors formed after inoculation of the hepatoma cells of the 5th year of cultivation. However, alpha-fetoprotein was not detected in the serum of mice with tumors induced by inoculation of the hepatoma cells of the 8th year of cultivation.


The effect of cultured endothelial cells on factor VIII procoagulant activity. Cultured human umbilical vein endothelial cells produce a protein that has von Willebrand factor activity and forms immunoprecipitates with rabbit antibody to purified plasma factor VIII/von Willebrand factor (FVIII/vWF) protein, but it has no FVIII procoagulant activity. Of the three characteristics of plasma FVIII/vWF protein, only FVIII procoagulant activity is readily destroyed by trace proteases. A previous report from this laboratory demonstrated protease activity in culture medium under conditions that had been used by others to show that endothelial cells do not synthesize protein with FVIII procoagulant activity. However, even if cultured endothelial cells are placed in protease-free culture medium, no FVIII procoagulant activity can be detected, despite an increase in the level of protein with vWF activity from 0 to 0.57 microgram/ml by 48 hr. This observation and the lack of protease activity in medium left in contact with the cells for 48 hr led to the hypothesis that proteases exist on the surface of cultured umbilical vein endothelial cells. Protease activity was quantitated by the hydrolysis of p-nitroaniline from the substrate, N-benzoyl-phenylalanyl-valyl-arginyl-p-nitroanilide and by degradation of the procoagulant activity of added purified plasma FVIII/vWF protein. In the absence of endothelial cells, no protease activity was present in protease-free culture medium whether or not it had previously overlaid cultured cells. This medium did not cause cleavage of p-nitroaniline from the tripeptide substrate, and 83% of added FVIII procoagulant activity remained after 48 hr. When the synthetic tripeptide was incubated in contact with cultured endothelial cells, 7.3 +/- 0.8 X 10(-10) moles of p-nitroaniline/hr was released; moreover, only 47% of the added FVIII procoagulant activity remained after 48 hr. Given this rate of destruction, it can be calculated that sufficient protease activity exists on the surface of cultured endothelial cells to degrade the procoagulant activity of approximately 1.6 microgram FVIII/vWF protein/hr. This degradation rate is 45 times the rate of release of FVIII/vWF protein from cultured endothelial cells when assessed by the generation of protein with vWF activity. Hence, the detection of FVIII procoagulant activity, if in fact synthesized by cultured endothelial cells, will be most difficult.


[Nuclear surface of buccal cells as a function of the nature of the stain. Measurements with an image analyzing computer]. Measure of nuclear areas was performed on buccal smears by the Quantimet 720 image analysing computer. Five different staining procedures were used and we consider that toluidin blue at pH 4 is the best for a fine quantitative caryometry.


[Modifications of the maternal immune response associated to pregnancy (author's transl)]. The author reviews the literature on the immune status of pregnant women. No major alteration of the immune response was observed. If any, the modifications are not important and involve the synthesis of immunoglobulins, the presence of circulating immune complexes, some sub-populations of mononuclear cells and the in vitro lymphocyte response to T dependent antigens. Phagocytosis, the number of B and T cells and mitogen lymphocyte response are normal. Studies on the immuno-suppressive effect of both pregnant women sera and serum substances (hormones, pregnancy associated or specific proteins, carcino-embryonic antigens) are reported. Discrepancies are observed between conclusions drawn by various authors: they relate to the efficiency of the measurement and to the observed effect. The contradictory results obtained with alpha-foetoprotein provide an illustration of the critical analysis of these studies. Finally, the in vivo immune status of pregnant women is reported as observed in cutaneous tests, transplantations and auto-immune diseases. The most relevant observations are: a defect of the expression of delayed hypersensitivity reaction, a facilitating effect of pregnancy in kidney transplantation similar to that obtained after transfusion and an inconstant and mild defect of graft immunity. Therefore, pregnancy produces a specific immune status about which tests presently available give few relevant results, similar to those obtained in some cancers.


Results of transurethral resection of prostate in patients with cerebrovascular disease. Twenty-two patients with bladder outflow obstruction and a past history of cerebrovascular accident are described. Sixteen patients regained normal micturition control. Poor results occurred in the more handicapped patients, especially in those with additional neurological problems. Patients who present with bladder outflow obstruction should be treated surgically. To determine those patients who will have a poor result full clinical and neurological assessment must be made, together with urodynamic studies. In order to improve these results a team approach is required between the urologist, geriatrician, nurse and physiotherapist.


A critical evaluation of the results of transurethral resection of the prostate. In a review of 1057 consecutive prostatectomies of which 95% were performed transurethrally, carcinoma was present in 11.8%. There were 10 deaths within a month of operation (0.9%), 9 of these patients having been exceptionally old and unfit. The rate of complications and the end results appear to justify using transurethral resection as the method of choice for prostatectomy whenever it is feasible.


A placebo-controlled double-blind study of the effect of phenoxybenzamine in benign prostatic obstruction. A double-blind placebo-controlled study of phenoxybenzamine for the symptomatic treatment of benign prostatic obstruction is reported. Statistically significant evidence of an improvement in both the peak and mean flow-rates was found. Both diurnal and nocturnal frequency were significantly diminished. Residual urine was unaffected, and the possible reasons for this are discussed. Urethral pressure recordings confirmed the reduction in the closure pressure in the prostatic segment of the urethra. It was concluded that there was good evidence that the treatment is effective.


Receptor proteins for androgens in prostatic disease. The suitability of an exchange assay with methyltrienolone (R1881) for the measurement of androgen receptors in human prostatic tumours has been investigated. Having established the specificity of this compound for androgen receptor assay, a pilot study was carried out to measure dihydrotestosterone (DHT) receptor in 27 cases of benign hypertrophy and 18 cases of malignant tumour of the prostate.


The value of alpha-fetoprotein and beta-human chorionic gonadotrophin in the management of testicular tumours. Serum alpha-fetoprotein and beta-human chorionic gonadotrophin levels were estimated in 115 patients. Sixty-nine patients had non-seminomatous germ cell tumours and 34 patients had seminomas. Twenty-six patients showed elevated serum makers.


A trial of bleomycin versus adriamycin in advanced carcinoma of the bladder. Thirty-seven patients were studied in a controlled clinical trial to assess the value of Adriamycin and Bleomycin in the treatment of advanced carcinoma of the bladder. The previously reported high response rates for both drugs were not observed in this trial. Administration of Bleomycin was accompanied by a toxicity rate so high that treatment had to be abandoned. Neither Adriamycin nor Bleomycin, therefore, used as single agents, have a role in the chemotherapy of advanced bladder cancer. Other agents should be investigated and assessed by controlled clinical trials that are disease-orientated rather than drug-orientated.


The Celestin tube in the palliation of carcinoma of the oesophagus and cardia. A series of 105 patients with dysphagia due to inoperable carcinoma of the oesophagus or cardia of the stomach, and 11 patients with dysphagia secondary to malignant medistinal nodes, was treated by intubation with a Celestin tube. The hospital mortality was high but 81 patients left hospital and 29 were still alive at 6 months. The prognosis for those with dysphagia due to malignant posterior mediastinal nodes was very poor, with no survivors at 6 months. Oesophageal perforation was a serious operative complication, with a hospital mortality of 83 per cent. Chest and wound infections were the most common postoperative complications occurring in 28 and 20 per cent of the patients, respectively. The restoration of swallowing was satisfactory in the majority of survivors although readmission with a blocked or displaced tube was not uncommon.


Does rectal examination affect serum acid phosphatase levels? Formol-stable serum acid phosphatase (SAP) was measured in 200 patients with symptoms of prostatism, before and at varying time intervals after digital rectal examination. In three separate groups of 50 patients SAP levels were measured before and at 5, 15 and 30 min following rectal examination and in a fourth group of 50 patients it was measured before and at 6, 24 and 48 h after examination. No significant change occurred in SAP levels following this examination in any of the groups studied. We conclude that rectal examination does not raise the SAP and that, contrary to popular belief, blood levels recorded at any time within this 48-h period are reliable.


Sudden death during ambulatory monitoring. Three patients with ischaemic heart disease died suddenly while being monitored with an ambulatory tape recorder. Two had terminal ventricular fibrillation initiated by paired bidirectional ventricular ectopic beats against a background of scattered ectopic activity; both had had ventricular tachycardia during routine treadmill exercise testing in the week before death. The third patient developed bizarre ventricular complexes followed by asystole. Sudden death may be due to ventricular fibrillation initiated by paired ventricular ectopic beats with changing morphology, or asystole following bizarre ventricular complexes. Exercise testing may have an important predictive value.


Analysis of fast axoplasmic transport in nerve ligation and adriamycin-induced neuronal perikaryon lesions. Fast axoplasmic transport was examined in experimental nerve ligation and adriamycin-induced neuronal perikaryon lesions. Animals were sacrificed at precisely 3 and 4 h following the injection of [3H]leucine. In single ligated nerves, the 3 and 4 h down-flow rates, which include the time for neuronal processing of labelled material, were 379.2 +/- 23.51 and 370.8 +/- 26.93 mm/day, respectively, compared to the calculated rate of fast axoplasmic transport of 345.6 +/- 25.22 mm/day. In double-ligated nerves the down-flow rates were 364.8 +/- 40.32 and 342 +/- 32.2 mm/day, respectively, while the fast axoplasmic transport rate was 273.6 +/- 36.16 mm/day. In contrast, in adriamycin toxicity the down-flow rates were 363.9 +/- 21.54 and 378 +/- 14.69 mm/day with a normal calculated rate of fast axoplasmic transport of 420 +/- 18 mm/day. These studies illustrate that evaluation of two (or more) different time intervals can be used to assess the site of impaired delivery of labelled materials to the peripheral portions of the nerve. Furthermore, a single time interval examination of transport should not be equated with axoplasmic transport especially under pathological conditions.


Autonomic block, cardiovascular depression and histamine release produced by polymyxin B in the cat. The present study quantifies the autonomic block, the cardiovascular depression and the histamine releasing effects of polymyxin B in nine anaesthetized cats. The dose requirements for 50 per cent depression of the mean arterial blood pressure, the bradycardiac response to vagal stimulation, and the contraction of the nictitating membrane elicited by pre-ganglionic and post-ganglionic stimulation of the cervical sympathetic trunk have a narrow range of scatter, being of the order of 6--12 mg.kg-1 of the polymycin B base. The neuromuscular blocking dose (ED50), previously determined and hereby confired, also falls in the same range. Bradycardia and possibly histamine release are also observable. The relatively unremarkable effect of polymyxin B on the heart rate can be attributed to the balance between the vagal, sympathetic and ganglionic blocks, as well as the possible histamine release. All effects are long-lasting. Thus lack of tissue specificity makes it implausible to explain the neuromuscular and the cardiovascular effects of polymyxin B solely by the cholinergic mechanism of action. Ganglionic block and histamine release do not completely explain hypotension induced by polymyxin B. We propose that all biological effects of polymyxin B derive from a common mechanism of action, which may be its antimembrane antibacterial action. We would like to alert anaesthetists to the possiblity that polymyxin B may cause severe multiple organ system depression, in addition to neuromuscular block.


A reliable Golgi-Kopsch modification.

A reliable Golgi-Kopsch modification. A reliable Golgi-Kopsch variant is described. Tissue is fixed with 1% paraformaldehyde, 1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.2. Tissue slabs (2.5--4 thick) are immersed for 5--7 days in a solution containing 3.5 g potassium dichromate, 15 g sucrose, 1.65 ml formaldehyde in 100 ml distilled water, washed briefly in 0.75% silver nitrate, and immersed in 0.75% silver nitrate for 2--3 days. Procedures for cutting, counterstaining, and handling sections are described.


The pathophysiology of asthma.

The pathophysiology of asthma. Because postmortem studies of humans provide little information on the initial pathophysiologic events in asthma, animal models have been developed. Recently the Ascaris-allergic rhesus monkey has provided an opportunity to examine the onset of pathophysiologic changes following challenge and to correlate them with airway structure. These studies have suggested that the initial interaction between antigen and mast cells may occur in the bronchial lumen or in the epithelium superficial to the tight junctions, where a small but significant percentage of airway mast cells exist. It also appears that this initial antigen-antibody interaction results in the release of mediators that both stimulate the rapidly adapting stretch receptors in the mucosa and alter the mucosal barrier so that proteins of large molecular weight can penetrate. The fact that antigen challenge results in hyperresponsiveness to a subsequent dose of inhaled histamine and increased systemic absorption of histamine suggests that the airway hyperresponsiveness could be related to increased penetration of histamine into the bronchial wall. These observations suggest that the initial event in an acute asthmatic attack is the release of mediators from superficial mast cells, and that this amplifies the allergic response by altering the mucosal permeability so that more antigen reaches the submucosal mast cells. This altered permeability may also help explain the hyperreactivity of the airways to nonspecific airway stimulants in persons with asthma.


Chemotherapy for advanced cervical cancer with methotrexate, bleomycin, and cis-dichlorodiammineplatinum(II). Nine patients with advanced cancer of the uterine cervix, having failed radiotherapy, were treated with a combination regimen of parenteral methotrexate, bleomycin, and cis-dichlorodiammineplatinum(II). Eight of the patients had objective partial remissions lasting a median of 4 months. All patients improved subjectively. Five of the patients, however, had sudden unpredictable onset of severe myelosuppression accompanied by stomatitis. Such frequent severe toxicity was not observed in patients with other malignancies treated with this combination. It is likely that impaired excretion of methotrexate resulting from ureteral dysfunction led to increased toxicity. In future protocols for advanced cervical cancer, methotrexate will be replaced by other effective agents. The very high response rate (89%) suggests that similar but less toxic combinations may contribute substantially to the future effective treatment of advanced cervical cancer.


Reversible effects of olfactory nerve section on behavior and biochemistry in mice. Coincident with the sectioning of the primary olfactory nerves in mice there is a dramatic loss of both olfactory-mediated behavior and the ability of the primary olfactory neurons to synthesize and transport the neuron specific marker, carnosine. These deficiencies are only temporary, and the ability of food deprived mice to locate buried food pellets returns within 21 days after olfactory nerve section. The ability of the primary olfactory neurons to synthesize and transport carnosine appears to increase between 8 days and 45 days after surgery. These data support the notion of behaviorally significant functional regeneration of olfactory neurons.


The early effects of chemical carcinogens on adult rat hepatocytes in primary culture. II. Effects on unscheduled DNA synthesis, cell division and alpha-fetoprotein production. The effects of exposure of rat hepatocytes in primary maintenance culture to chemical carcinogens has been studied with respect cytotoxicity and alterations in mitotic index, unscheduled DNA synthesis and alpha-fetoprotein (AFP) production. All compounds tested produced cytotoxicity. Increases in mitotic index and unscheduled DNA synthesis and the production of AFP were observed after treatment of the cells with the carcinogens but not after treatment with the non-carcinogenic isomers. These increases were dose-dependent and depended on the time of exposure and the time incubated postexposure. The patterns of the increase in mitotic index and AFP production after cessation of carcinogen exposure were very similar, with the increase in mitotic index occurring slightly before that for the AFP production and it is suggested from this and other data that the production of AFP is dependent on the generation of a cell species functionally distinct from the non-dividing hepatocytes. It is also suggested that measurement of unscheduled DNA synthesis in conjunction with that of AFP production in cultured hepatocytes may be useful as part of a screening programme for chemical carcinogens.


[Demonstration of antigen associated with human breast cancers]. The insoluble pellet of human mammary carcinomas was solubilized by an acid buffer. Antiserum prepared with this acidosoluble fraction, after suitable absorption gave one precipitin line with the immunizing extracts: this line is different from those given by the tumor associated antigens actually known. The same antiserum reacted only with sections of human mammary carcinomas by immunofluorescence . It did not stain sections of normal mammary glands or benign mammary diseases. Reactivity with cancers of other organs was absent or doubtful. Hence it is likely that an antigen associated to human mammary carcinomas was characterized.


[Heparin inhibition of the antithrombin activity of alpha-2-macroglobulin]. The interaction between thrombin and alpha-2-macroglobulin was studied on human purified materials, either in the presence or in the absence of heparin, by kinetic analysis of thrombin inhibition and polyacrylamide gel electrophoresis. In the absence of heparin, binding of thrombin to alpha-2-macroglobulin, shown by electrophoresis, leads to the loss of the coagulant property of the enzyme. In the presence of heparin the rate of inhibition of thrombin clotting activity by alpha-2-macroglobulin is strongly decreased. Heparin binds to thrombin, impairing the formation of thrombin-alpha-2-macroglobulin complex. These data show that heparin paradoxically protects thrombin from inhibition by alpha-2-macroglobulin whereas it increases the enzyme inhibition by antithrombin III. Such a phenomenon could be of practical interest for treatment of thrombosis in patients with high plasma level of alpha-2-macroglobulin and low level of antithrombin III, such as occurs in the nephrotic syndrome.


Radioimmunoassay of "pregnancy-specific"-beta 1-glycoprotein (SP1). A radioimmunoassay for SP1 ("pregnancy-specific" beta 1-glycoprotein) is described in which a sensitivity of 0.1 microgram/L was achieved by use of an antiserum with Ka approximately equal to 1.7 x 10(11) L/mol and delayed addition of a 125I-SP1 tracer (specific acty, 25 Ci/g) that was stable for use in the assay for at least six to seven weeks. We also report an additional anti-SP1 serum, from an early bleeding, that displayed "apparent positive cooperativity" in the IgG but not the IgM fraction.


A new and universal free/bound separation technique for the "CENTRIA" automated radioimmunoassay system. A new separation procedure based on the double-antibody technique has been adapted to the CENTRIA System. This procedure is universally applicable and lends itself to easy adaptation to commercial RIA kits in which liquid reagents are used. This second antibody is covalently linked to agarose (Sepharose) and the lyophilized powder is subsequently tableted for easy use on the instrument. The technique was applied to radioimmunoassay for thyrotropin (thyroid stimulating hormone), alpha-fetoprotein, and ferritin. Performance characteristics were as follows: sensitivity 1.5 milliunits/L, 4.5 micrograms/L, and 4.5 micrograms/L, respectively; intrarun precision 1.7, 9, and 3.4%, and interrun precision 7.6, 13, and 14.5%. All three assays were clinically validated.


Induction of human antigen-specific and non-specific helper factors in vitro. Human peripheral blood leucocytes, cultured for 4--6 days with the appropriate concentration of protein or synthetic polypeptide antigen, yield helper cells. These cells, after a further 24 hr incubation with the appropriate antigen release helper factors, which are of two types, antigen specific and non-specific; a similar situation to that described in the mouse. These factors are assayed by their effects on the plaque-forming cell response of mouse spleen cells cultures in vitro. The potential applications of this new assay for human T cell function are discussed.


beta 2-Microglobulin: methods and clinical applications. beta 2-Microglobulin is a low molecular weight protein that is found in most biological fluids. It was originally isolated from urine of cadmium-poisoned patients. Its amino acid sequence was established and shown to be structurally related to immunoglobulin constant domains. With the aid of antibodies specific against beta 2-microglobulin, the protein was detected on the membranes of all nucleated cells, normal and neoplastic. Measuring the quantity of beta 2-microglobulin showed that high levels are present in patients with renal tubular deficiencies and several other pathological conditions including neoplastic diseases. Extremely high levels were detected in seminal fluid and colostrum. Despite the structural relationship to immunoglobulins, no immunological relationship was demonstrated with these proteins using antibodies specific for beta 2-microglobulin. However, such antibodies are cytotoxic to all cells carrying beta 2-microglobulin on their surfaces. The discovery that beta 2-microglobulin is an integral part of the histocompatibility antigens of human and murine origin stimulated further research and interest in this molecule. Several groups of investigators have shown that beta 2-microglobulin is the low molecular weight chain and is noncovalently bound to a high molecular weight chain which carries the histocompatibility antigens. The structure of the histocompatibility antigens of lymphocytes (HLA) was shown by immunochemical as well as biological methods, and it is now well accepted. The antibodies against beta 2-microglobulin are extremely useful in the isolation of the histocompatibility antigens for sequence studies. Furthermore, the antibody to beta 2-microglobulin revealed that other structures may be bound to beta 2-microglobulin such as phytohemoagglutimin (PHA) receptors, mixed lymphocyte culture (MLC) antigens, etc. Murine thymus leukemia (TL) antigen also contains beta 2-microglobulin as an integral part of its structure; other tumor antigens may have a similar structure. Through all these studies, beta 2-microglobulin emerged as the best known membrane protein that can serve as a model for study of the arrangement and the function of the cell membrane.


Subcellular localization of polluting metals in roadside earthworms exposed to traffic exhaust gases. The aim was to make a subcellular localization of metals in tissue from lumbricid earthworms exposed to environmental pollution. Scanning transmission electron microscopy in combination with energy dispersive X-ray microanalysis of tissue fixed in glutaraldehyde only, and with no electron staining, was used. Zinc was registered in the metachromatic mucous granules of the epidermal cells, and zinc, iron and lead in the chloragosomes of the chloragocytes suggesting that metal may be excreted together with the slime or stored in chloragosomes. Relatively few metal nuclear inclusions were encountered probably due to the fact that some metal leaks out during the preparation process. A comparison is made with a chemical analysis of cellular fractions (Talberg, 1977).


The role of chromosomal proteins in the C-banding of Allium cepa chromosomes. When chromosomes of Allium cepa are subjected to a C-banding procedure (incubation in saturated barium hydroxide followed by phosphate buffer at 60 degrees C for 1 h) and then treated with Giemsa stain, bands appear at the telomeres of all chromosomes. Microspectrophotometric measurements of Feulgen-DNA content, demonstrated that the C-banding procedure extracted DNA from the nuclei. Staining of banded chromosomes with several DNA-specific stains showed that this loss was differential, with the band DNA exhibiting more resistance to extraction than that of the rest of the chromosome. The C-banding procedure did not extract chromosomal proteins, however, and no difference in mass per unit length could be detected by Nomarski optics between band and interband regions. Several experiments demonstrated that chromosomal proteins play a significant role in C-banding. First, treatment of chromosomes with pronase before C-banding resulted in the elimination of differential staining with Giemsa. Furthermore, in preparations where the DNA was completely hydrolysed with hot TCA, the remaining chromosomal proteins were found to exhibit a differential affinity for Giemsa stain. Amido black staining demonstrated that total chromosomal protein was uniformly distributed after the hot TCA digestion, but the proteins localized in the telomeres had a greater affinity for the Giemsa stain than the bulk of the chromosomal proteins. When the TCA-digested chromosomes were subjected to the C-banding procedure before staining, the differential affinity of the telomeres for the Giemsa stain was lost. Thus, C-banding appears to be the result of a complex interaction between protein and DNA in which the greater resistance to extraction of the band DNA is necessary to stabilize and preserve chromatin protein which exhibits a differential affinity for Giemsa stain.


[Conservative treatment of echinococcus alveolaris of the liver: preliminary observations (author's transl)]. Mebendazol (Vermox) was given for six months at an oral daily dose of 2 g to three patients with inoperable infestation of the liver by Echinococcus alveolaris. In two patients the general state improved greatly, with reduction in the size of the liver infiltrate, regression in serum bilirubin and alkaline phosphatase levels. Treatment continues in the third case.


Binding specificity of rat alpha-fetoprotein for a series of estrogen derivatives: studies using equilibrium and nonequilibrium binding techniques. The binding specificity of rat alpha-fetoprotein (AFP) for a series of estrogen derivatives has been investigated and compared to that of the uterine cytosol estrogen receptor. In addition, several equilibrium and nonequilibrium binding techniques have been examined for their appropriateness in studying AFP-estrogen interactions under different experimental conditions. The ratio of association constants (RAC) of various estrogen derivatives (compared to 17 beta-estradiol; RAC = 100%) were determined using a rapid and convenient miniaturized equilibrium assay that utilizes the partitioning properties of Sephadex G-25. AFP has the highest affinity for estrone derivatives and is especially tolerant of small, nonpolar substituents in the D ring; it binds nonsteroidal estrogens and antiestrogens weakly or not at all. Thus, it has a binding specificity quite unlike that of the estrogen receptor. Several potential affinity labels were studied. Of these, 16-diazoestrone, a photoreactive compound, has the highest affinity for AFP (RAC = 121%). Another equilibrium technique, steady state polyacrylamide electrophoresis, effects a clear separation between AFP and albumin. It is superior to conventional polyacrylamide electrophoresis under conditions where dissociation is rapid. Charcoaldextran adsorption also rapidly removes estradiol that is bound to AFP (t1/2 = approximately 10 min at 0 C). The AFP-estradiol complex appears to be stabilized by adsorption to hydroxylapatite, however, so that hydroxylapatite can be employed in a batchwise manner to assay binding to AFP. This is the most appropriate technique when high concentrations of ligand are used. These studies highlight the marked differences in binding specificity between rat AFP and the uterine cytosol estrogen receptor and indicate the techniques most appropriate for binding measurements under different conditions. This work should facilitate the design of affinity labeling reagents for rat AFP as well.


Functional cytology of the pituitary gland of the teleost Clarias batrachus (L.). The RPD (rostral pars distalis) is formed of two cell types. The erythrosinophilic acidophils were hypertrophied in response to ammonium sulphate treatment and dark background adaptation. The response to former may be due to osmoregulatory involvement. The PbH positive corticotrophs were markedly hypertrophied and degranulated in response to metopirone and ammonium sulphate treatments. They were moderately activated in the dark and white background adapted fish. The PPD (proximal pars distalis) is composed of two types of cyanophils and acidophils. The thyrotrophs are identified by their hypertrophic response to radiothyroidectomy and thiourea treatment. They are distributed in the ventral part of the PPD, posterior neurohypophysis and PI (pars intermedia). The gonadotrophs which largely occupy the dorsal aspect of the PPD increase numerically as the gonads mature and become the major cell type before spawning. The acidophilic somatotrophs are distributed all over the PPD. They are the predominant cell type in the PPD of fish with immature gonads. They gave strong reaction to Baker's acid haematein indicating the presence of phospholipid. Both PbH and PAS cells of PI were stimulated in fish kept in dark background whereas they did not exhibit any obvious change under white background adaptation.


Disposition of the pulmonary toxin, bleomycin. The disposition of bleomycin in rats was investigated with the use of a 57Co-labeled bleomycin radioimmunoassay. Bleomycin (1 mg/kg, ip) reached a peak serum level of 0.7-1.0 microgram/ml in 54 +/- 4 min (mean +/- SE) and was eliminated with a t 1/2 of 32 +/- 1 min. The apparent volume of distribution in rats was 184 +/- 11 ml/kg. No change in t 1/2 was observed when rats were pretreated with phenobarbital (1 mg/ml in drinking water for 4 days) or exposed to ozone (3 ppm, 24 hr). Probenecid (0.2 g/kg, ip) given 20 min prior to bleomycin administration increased the t 1/2 2-fold. The order of tissue distribution of 57Co-labeled bleomycin 2 hr after ip administration was: kidney greater than liver greater than lungs greater than spleen greater than heart greater than thymus. The affinity of 57Co-labeled bleomycin for homogenates of lungs, liver, or kidney was equal.


Inhibition of cortisone action in mice by heparin. A single dose of heparin applied in a depot-form (Freund's incomplete adjuvant or Ca-phosphate gel) inhibits the effects of intraperitoneally injected cortisone on the lymphoid organs (thymus and spleen), on the peritoneal and peripheral lymphoid cell count and serum gamma globulin level as well as on the liver glycogen deposition in mice. The same dose of heparin did not influence the action of hydrocortisone measured on the thymic and spleen involution and liver glycogen content. The route of cortisone administration seems to be critical, as heparin shows no or only minor effects when cortisone is applied subcutaneously; moreover, the action of cortisone per se is more marked by subcutaneous than by intraperitoneal administration. The results suggest the hypothesis that heparin inhibits cortisone-cortisol conversion and this inhibition is mediated by macrophages.


Immunochemical studies of beef pancreas tryptophanyl-tRNA synthetase and its fragments. Determination of the number of antigenic determinants and a comparison with tryptophanyl- tRNA synthetases from other sources and with reverse transcriptase from avian myeloblastosis virus. The immunoglobulin G (IgG) fraction of the antiserum from rabbits immunized with homogeneous beef pancreas tryptophanyl-tRNA synthetase inhibits the enzyme activity in the reactions of both tRNATrp aminoacylation and tryptophan activation. Fab fragments of IgG act in a similar way. Common antigenic determinants have been detected in tryptophanyl-tRNA synthetases from beef, pig, chicken and rat livers using pure antibodies against beef pancreas tryptophanyl-tRNA synthetase. This observation indicates the evolutional stability of certain structural features of tryptophanyl-tRNA synthetases. The interaction of antibodies with the fragments of beef tryptophanyl-tRNA synthetase produced by endogenous and tryptic proteolysis of the enzyme has been studied. On third of the antiserum antibodies interacting with the C-terminal fragment of the enzyme (Mr approximately equal to 40000) inhibits its activity whereas the antibodies to the N-terminal fragment (Mr approximately equal to 20000) have no effect on the enzyme activity. The immunochemical identity of the two synthetase fragments differing in their enzymatic activity supports the assumption that the loss of enzymatic activity of the tryptic fragment is caused by lack of a small peptide which is retained in case of endogenous proteolysis; probably the amino acid residues of this peptide participate in formation of active centre of tryptophanyl-tRNA synthetase. A radioimmunochemical method is described for determining the number of antigenic determinants. One molecule of tryptophanyl-tRNA synthetase was found to bind 9 (+/- 1) molecules of Fab fragments. Antibodies against tryptophanyl-tRNA snythetase from beef pancreas do not inhibit noticeably the activity of reverse transcriptase from avian myeloblastosis virus. No antigenic determinants in common have been detected in reverse transcriptase and tryptophanyl-tRNA synthetase by radioimmunochemical assays.


Structural studies on the immunodominant group of lipid A from lipopolysaccharide of Yersinia pseudotuberculosis. Lipid A isolated from lipopolysaccharide of Yersinia pseudotuberculosis was used for immunization of rabbits to afford antisera to lipid A with titers of 1:640 in the passive hemolysis test. Exhaustion of immune serume with sheep erythrocytes decreased antibody titers up to 1:160. Authentic samples of 2-(DL-3-hydroxytetradecanoyl)amino-2-deoxy-D-glucose 6-phosphate, 2-tetradecanoylamino-2-deoxy-D-glucose 6-phosphate and 2-acetamido-2-deoxy-D-glucose 6-phosphate have been synthesized in order to carry out a comparative study of inhibitory activity of these compounds and lipid A using a system of lipid A and antiserum to lipid A. As a result, the immunodominant moiety of the lipid A of Y. pseudotuberculosis proved to contain a D-glucosamine residue acylated with 3-hydroxytetradecanoic acid at the amino group. The nature of the fatty acid acylating the amino group of glucosamine does not play an important role in the structure of immunodominant moiety of lipid A.


Immune response deficiency of BSVS mice. II. Generalized deficiency to thymus-dependent antigens. BSVS mice gave abnormally low IgG responses to 5 thymus-dependent antigens as well as a weak delayed-type hypersensitivity (DTH) response to sheep red blood cells. In contrast to IgG, the IgM antibody responses of these mice were normal to three T-independent antigens as well as to all five T-dependent antigens. The low immune responsiveness of BSVS mice was also reflected in the low levels of IgG(2)a, IgG(2)b and IgG(3) in their normal serum. The low T-dependent immune responses may result from BSVS mice having been selectively bred for susceptibility to infection with St. Louis encephalitis virus and Salmonella. C57BL/6J mice, which are also highly susceptible to Salmonella, gave low immune responses similar to, but genetically distinct from, those of BSVS mice. The levels of Ig-positive and theta-positive cells were normal in BSVS and C57BL/6J mice.


In vitro induction of delayed-type hypersensitivity. Murine spleen cells, cultured in vitro for 6 days in the presence of high concentrations of burro erythrocytes (BRBC), are sensitized to exhibit delayed-type hypersensitivity (DTH) specific for this antigen. Such cells, on being injected with antigen into the footpads of normal mice, cause a 24-h swelling reaction. This activity of the cultured cells requires the presence of BRBC both during the in vitro incubation and in the footpad. The activity of the sensitized cells in causing swelling is sensitive to anti-Thy-1 antibody and complement, and the kinetics of the swelling reaction are characteristic of a DTH response. In vivo low-dose priming of the spleen cell donors considerably enhances the ability of the cultured cells to cause swelling. This system provides a means of studying the regulation of the induction of DTH in vitro.


The central cervical nucleus in the cat. II. The cerebellar connections studied with retrograde transport of horseradish peroxidase. After HRP injections in kittens and adult cats into parts of the cerebellum known to receive spinal afferents, retrogradely labelled neurones were found in the C1--C4 segments of the spinal cord, primarily in the central cervical nucleus (CCN). A few labelled neurones were also found in laminae IV and VI--IX of Rexed. The results obtained in kittens and adult cats were similar. Control HRP injections were made into the vestibular nuclei and the inferior colliculus. After injections mainly involving the vestibular nuclei a few faintly labelled neurones were found in the C1--C4 segments in, and occasionally also outside the CCN, in laminae VI--VIII. After injections into the inferior colliculus, however, no labelled neurones were observed in these segments. After spinal cord hemisections at C1 and bilateral HRP injections into the anterior lobe, nearly all labelled CCN neurones were found ipsilateral to the hemisections. Labelled axons of such CCN neurones could be traced across the midline to the lateral funiculus. A few neurones were found outside the CCN in laminae VII--IX. Contralaterally labelled neurones were found mainly in lamina VI.


The central cervical nucleus in the cat. III. The cerebellar connections studied with anterograde transport of 3H-leucine. Injections of 3H-leucine were made into the region of the central cervical nucleus (CCN) in the C1--C4 segments of the spinal cord in 19 adult cats. In the cerebellum labelled mossy fibre terminals were found bilaterally concentrated in lobule I and adjoining parts of lobule II of the anterior lobe. In addition, a fair number of terminals were found in the basal parts of lobules III--VIII. The terminals were mainly found in the vermal zone. Only a minor proportion was observed in the intermediate zone of the anterior lobe and only occasionally were terminals seen in the hemispheral parts. No labelled climbing fibres were observed. Axons probably derived from the CCN were found to ascend in the brain stem, mostly contralateral to the injection site. They passed close to the vestibular nuclear complex and some fibres appeared to terminate in the nucleus x of Brodal and Pompeiano. The majority of axons entered the cerebellum via the superior cerebellar peduncle, a few via the restiform body. The findings suggest that lobules I--II are important recipients of information from the neck.