Mannitol oxidation in two Micromonospora isolates and in representative species of other actinomycetes. Mannitol kinase and mannitol-1-phosphate dehydrogenase activities were detected in two Micromonospora isolates. The presence of these enzyme activities indicates that mannitol is catabolized first to mannitol-1-phosphate and then to fructose-6-phosphate. Mannitol-oxidizing enzymes were also surveyed in representative species of four other genera of actinomycetes. Mannitol-1-phosphate dehydrogenase was detected in cell-free extracts of Streptomyces lactamdurans. In contrast, cell-free extracts of Mycobacterium smegmatis, Nocardia erythrophila, Streptomyces lavendulae, and Actinoplanes missouriensis contained mannitol dehydrogenase activity but no detectable mannitol-1-phosphate dehydrogenase activity. The mannitol dehydrogenase activities in the latter species support the operation of a pathway for catabolism of mannitol that involves the oxidation of mannitol to fructose, followed by phosphorylation to fructose-6-phosphate.


Comparison of sulfite-polymyxin-sulfadiazine medium and tryptose-sulfite-cycloserine medium without egg yolk for recovering Clostridium perfringens. The overall recoveries of spores and of actively growing, heat-stressed, coldshocked, and frozen cells of five strains of Clostridium perfringens were significantly greater (95% confidence limits) on tryptose-sulfite-cycloserine medium without egg yolk than on sulfite-polymyxin-sulfadiazine medium.


Glucagonoma syndrome. Report of two cases and literature review. The glucagonoma syndrome is characterized by dermatitis, stomatitis, elevated serum glucagon levels, abnormal glucose tolerance, weight loss, and anemia--all in association with a glucagon-secreting alpha-cell tumor of the pancreas. A review of 21 cases showed strikingly similar features. A generalized, symmetrical dermatitis initially appeared to be asteatotic or eczematous over the perineum, buttocks, and lower extremities. Gradually, a more characteristic migratory necrolytic erythema with transient bulla formation and erosions developed in intertriginous and dependent areas. Histologically, the most specific features included necrolysis of the upper epidermis, with liquefaction necrosis of the granular cell layer and subcorneal clefting or blister formation. The dermatologist is often first to examine such patients; early recognition of this syndrome with prompt surgical removal of the primary pancreatic lesion may afford cure of the neoplasm.


Necrolytic migratory erythema. Distinctive dermatosis of the glucagonoma syndrome. Glucagon-secreting tumors of the pancreatic islets (glucagonomas) produce a distinctive syndrome in which weight loss, diabetes mellitus, anemia,and prominent mucocutaneous findings occur. The cutaneous component-necrolytic migratory erythema--may be polymorphous, but most commonly manifests as erosions and crusts of the groin, perineum, buttocks, distal part of the extremities, and central area of the face. Alternatively, scaly papules and plaques may predominate in these areas. The eruption may resemble such dermatoses as pemphigus foliaceus, acrodermatitis enteropathica, chronic mucocutaneous candidiasis, psoriasis, and severe seborrheic dermatitis. Two patients with chronic, previously undiagnosed dermatoses had necrolytic migratory erythemia, which led to the discovery of glucagonomas present in each. In one patient surgical resection of the tumor resulted in total clearing of the rash within 48 hours. Awareness of this distinctive entity may lead to early diagnosis and, possibly, cure.


Generalized acute mucocutaneous herpes simplex type 2 with fatal outcome. To our knowledge, we present the first culturally documented case of generalized acute mucocutaneous herpes simplex type 2 in an immunosuppressed adult who died during the course of the disease. The picture of the clinical spectrum of herpetic disease in immunosuppressed patients is detailed, and the need for typing the virus, particularly in the disseminated form, and for clarifying possible differences in epidemiology, clinical expression, and prognosis is emphasized.


Perinatal cytomegalovirus infection in man. In a prospective study of 148 children from urbanized southern Finland 3 were found to be congenitally and 48 perinatally infected with cytomegalovirus (CMV), while 6 developed "late" infection during the first year of life. During pregnancy and the first year after delivery 23 of the mothers had no CMV antibodies; none of the children of these seronegative mothers developed any type of CMV infection. Fresh blood exchange transfusions did not increase the risk of CMV infection. The data support the hypothesis that the mother is the source of perinatal CMV infection. Children with a low birthweight not due to prematurity, and first children seem to run a greater risk of acquiring perinatal CMV infection. If the child is breast fed up to the age of 2 months the risk seems to be increased. Perinatal CMV infection gave rise to no symptoms or signs and had no effect on growth or on motor and psychosocial development during the first year of life.


Lack of effect of phototherapy on plasma cyclic-AMP in newborn infants. The effect of phototherapy on plasma cyclic-AMP in newborn infants has been studied. Our results suggest that phototherapy does not produce appreciable changes in plasma cyclic-AMP levels.


Thoracic neurolophomas.

Thoracic neurolophomas. Neural crest cells are unique. Few tissues found in the embryo are expressed with such diversity in adults. Cellular differentiation occurs simultaneously with widespread migration. Neural crest cell neoplasia results in some of the most common tumors clinically encountered. As a group, these tumors are called neurolophomas. This report describes the neural crest formation, the tissues normally derived from it, a classification of the neural crest tumors, and a review of such tumors as they arise in the thorax.


The effect of enkephalin and enkephalin analogs on the guinea-pig ileum and rat brain opilate receptor. We have synthesized the two naturally occurring enkephalins (Methionine-Enkephalin and Leucine-Enkephalin) as well as four other analogs (Methinonine-Enkephalin amide, S-Benzyl sulphonium analog of Methionine-Enkephalin, 3-Benzyl-tyrosine analog of Methionine-Enkephalin and 3-Benzyl-tyrosine analog of Leucine-Enkephalin) in order to study their relative potency and their inactivation of different tissues. Morphine-like activity of the peptides was determined by measuring their inhibitory effect in the guinea-pig ileum and their interaction with the brain opiate receptor; the order of affinity of the three most active peptides (Methionine-Enkephalin amide greater than Methionine-Enkephalin greater than Leucine-Enkephalin) for the rat brain opiate receptor was found to be the same in the guinea-pig ileum. The rate of inactivation of the peptides was also studied in the guinea-pig ileum and in the rat brain membrane preparation. Methionine-Enkephalin amide was relatively more resistant to the ileum peptidases than the other peptides, but was destroyed at the same rate by brain membranes; this variation in inactivation of Methionine-Enkephalin amide probably indicates the presence of differnet peptidases in each tissue preparation.


Salt-wasting bronchorrhea and its mechanisms. A 56-year old woman, with the diffuse form of bronchioloalveolar cell carcinoma, developed massive bronchorrhea, resulting in severe fluid and electrolyte depletion when her oral intake was compromised. Chemical analysis of the bronchial secretions and the ultrastructural features of the tumor cells support the concept of an active secretory process. The lactic dehydrogenase (LDH) isoenzyme pattern of the fluid is similar to that found in fetal cells. The neoplastic cells may acquire a more primitive LDH pattern.


[Albright's type II osteodystrophy and hypothyroidism due to TSH deficiency. Apropos of a case in a child]. The report is dealing with a 10 year-old girl. The diagnosis of Albright's type II osteodystrophy relied on clinical, radiological and biological symptoms: evocative dysmorphic syndrome and absence of disturbances in the phosphocalcic metabolism. Hypothyroidism was secondary to an isolated defect in TSH, and a TRH stimulation test did not induce an increase in plasma TSH.


[A symptomatic parathyroid adenoma. Value of parathyroid hormone determination through selective catheterization of the thyroid veins]. A parathyroid adenoma is reported in a girl aged 12 years in whom hypercalcaemia was discovered by chance. Investigation of calcium metabolism suggested the diagnosis of hyperparathyroidism and studies of the urinary cyclic AMP and determination of the plasma parathyroid hormone concentration further added to the evidence. The diagnosis of parathyroid adenoma was made after determination of the parathyroid hormone concentration at various sights during selective catheterization of the tyroid veins. This was confirmed at surgery. In this patient the place of catheterization of the inferior thyroid veins in the early diagnosis of primary hyperparathyroidism is discussed.


[Islet cell adenoma with neonatal onset. Clinical, hormonological and ultrastructural study of a case]. A case of hyperinsulinism occuring in a newborn, with a birthweight of 4,050 g, is reported. The hypoglycaemia was refractory to the usual therapy (increase of glucose administration per os, and I.V., corticosteroids, glucagon, diazoxide). At surgery, undertaken at 9 days of age, an adenomatous nodule was removed along with the left part of the pancreas. Death occurred at 18 days, after the child had developed a transitory acidoketosic diabetes and an encephalopathy. Measurement of insulin by radio-immunoassay revealed a strong increase in the ratio insulin/glycaemia, characteristic of nesidioblastoma, as well as a high concentration of insulin in the tumor as compared to normal tissue. On the ultrastructural level, the observed features differed from those seen in children and adults and showed an abnormal overload of dense deposits in the cytoplasm of some histiocytes.


Family intervention with severely disturbed inpatients. Although there is considerable evidence that family variables and interaction patterns affect the duration of hospitalization and frequency of readmission of psychiatric patients, it is rare to find inpatient facilities having psychotherapeutic programs that systematically stress family interventions. This article discusses the difficulties encountered by both families and staff as they cope with the issues of hospitalization, suggests a structured, patient-oriented family approach, and recommends some specific techniques for short-term work with families of severely disturbed inpatients.


Invasion of malignant cells into cultured embryonic substrates. To study malignant invasion, we associated tissue and cell culture fragments, with adhesive and non-adhesive living substrates in vitro (11). Non malignant mesonephros, quail heart and BHK-cells, as well as malignant HeLa-, Hepatoma-, Harding-Passey melanoma-, Py-, TLX5 lymphoma- and Schmidt-Ruppin sarcoma cells, were transplanted into cultured organ fragments of chick embryos and chick blastoderms. Malignant invasion was evaluated on the basis of the following histological criteria: 1. changes in organization of the graft. 2. infiltration of cells from the graft into the substrate, 3. degenerative alterations of the substrate. It was shown that adhesion of the graft to the substrate is a prerequisite for malignant invasion. Invasion into non-adhesive substrates, such as the apical side of epithelia, was never observed. Contrary, all malignant cells did invade into adhesive substrates. Interposition of a vitelline membrane always inhibited the expression of invasiveness. The morphological pattern of invasion depended mainly on the architecture of the substrate and differential resistance of its various components explained well all histological pictures. From the latter observation and from the localization of degenerative changes in the substrate we inferred that malignant cells exert their deleterious effect most probably upon immediate contact with their host.


The role of cyclic adenosine 3',5'-monophosphate (cAMP) in the mechanism of action of tricyclic antiderpressants. In the course of investigations on experimental arterial hypertension in rats, it has been stated, that imipramine prevents the development of hypertension and that is simultaneously causes an accumulation of cAMP in the vessel walls. The relationship between the two phenomena has been discussed.


Degeneration of aortic smooth muscle cells in swine fed excess vitamin D3. Twenty-eight weanling swine were divided into two groups and fed a basal ration of corn and soybean meal or one containing a high fat and cholesterol supplement. One subgroup of each received 220,000 IU of vitamin D3 per kilogram. Grossly normal areas of the distal abdominal aorta were used for quantitative comparison of degenerated smooth muscle cells by electron microscopical cell counts. Degeneration was classified into rarefaction, condensation, and frequency per 100 nucleated cells. The frequency of dead cells was demonstrable between the swine fed excess vitamin D3 with or without high fat and cholesterol and those fed the basal ration alone, or frequencies of 7.9, 7.4, and 5.6 at 3 months of age (P less than .05) and 7.3, 6.2, and 5.1 at 6 months (P less than .05), respectively. No significant difference was demonstrable between the swine fed high fat and cholesterol without excess vitamin D3 and those fed the basal ration alone, or frequencies of 5.7 and 5.6 at 3 months of age and 5.5 and 5.1 at 6 months.


Interferon production and response to exogenous interferon in two cell lines of mouse brain origin persistently infected with Sendai virus. Two persistently infected cell lines established from C3H mouse brain cells infected in vivo with Sendai virus were shown to differ with respect to interferon (IF) production and response to exogenous IF. MB/Sen carrier cells contained 1-5 per cent antigen positive cells when examined by immunofluorescence, and virus was occasionally recovered from the culture medium. MB/SenAS carrier cells were maintained with 0.16 per cent Sendai antiserum in the supernatant medium. All MB/SenAS cells contained viral antigen and infectious virus was present in the culture medium. MB/Sen released IF spontaneously into the culture medium. Further IF production could be stimulated in MB/Sen by superinfection with Newcastle disease virus (NDV) or vesicular stomatitis virus (VSV). Exogenous IF provided good protection against VSV challenge. In contrast, MB/SenAS produced no IF spontaneously but could be stimulated by NDV and VSV to produce IF. Exogenous IF failed to reduce the amount of VSV released into the supernatant fluid. Replication of VSV was restricted in MB/SenAS as shown by a 2.3 log10 lower virus yield compared to MB/Sen.


Components of parainfluenza-3 virus, SLP-strain, reacting in assays of cell-mediated immunity in cattle. Parainfluenza-3 virus was isolated by affinity chromatography, including a purification step with immobilized lectin Vicia erwilia. The peplomers of disintegrated virus were similarily isolated using another carbohydrate-specific lectin Vicia crotalaria. The whole virion and the peplomers were both active as antigens in the leucocyte migration inhibition, lymphocyte stimulation and skin hypersensitivity tests. The remaining virus material, freed of detergents used for virus disintegration and containing nucleocapsids, did not act as antigen in these tests of cell-mediated immunity.


[Enzymomorphology of the vaccinal process in connection with formation of immunity to plague]. The studies conducted showed that the beginning of immunogenesis following the administration of live plague vaccine was preceded by a period of primary toxic action of multiplying microbes when the activity of a number of enzymes in the organs of guinea-pigs was temporarily reduced. Subsequently, inductive and productive phases of formation of antibodies started against the background of normalization and growth of the enzymatic activity associated with mobilization of energy. The earliest hyperplastic changes during immunogenesis occurred in the thymus-dependent areas of the lymphoid organs. Somewhat later, hyperplasia of B-structures dependent on the bone marrow started to prevail.


[Activity of different antigenic preparations from the retina to induce experimental auto-immune uveo-retinitis (EAU) in guinea pigs (author's transl)]. 24 different antigenic preparations from bovine or guinea pig retina and 3 from bovine uvea were tested for their ability to induce uveo-retinitis in guinea pigs. Each animal received one injection into the hind foot pads of 0.1 ml og the tissue preparation mixed with an equal volume of complete Freund's adjuvant. The intensity of the disease was assessed by clinical and histological criteria. Homogenates and extracts from whole guinea pig retina are more active than the same preparations from bovine retina. Autologous retinal extract is slightly more active than homologous in low doses. In bovine retina, the autoantigen(s) is localized in the photoreceptor structures and the pigment epithelium. Bovine uveal preparations seem to be inactive when the epithelium has been removed. Purified outer segments are very active, as well as soluble extracts of outer segments. Highly purified bovine rhodopsin has no immunopathogenic activity. A soluble autoantigen (autoantigen S) has been isolated by preparative isoelectrofocusing from retinas of several species. Autoantigen S from guinea pig induces the disease in guinea pigs at a dose of a few micrograms.


[Relaps of a nasopharyngeal fibroma after 20 years (author's transl)]. In a 38 year old patient there was a relapse of a nasopharyngeal fibroma in the buccal mucous membrane which had been operatively removed from the nasopharynx 20 years ago. Both tumors were histologically identical. The patient did not suffer from hormonal disturbances.


Malignant disease in childhood: the problems in general practice. The role of the family doctor with regard to malignant disease in childhood, can be summarized as follows: To act appropriately so that early diagnosis, which to an important degree determines the prognosis, is not delayed by a justifiably low index of suspicion. To assist the family, including the siblings, to cope with the severe emotional stress inherpent in the diagnosis of a malignancy in a child. To recognize the extreme danger of chicken pox and measles when contracted during chemotherapy, for cytotoxic drugs are also immune-suppressants, and appropriate immune globulins can be life-saving. To arrange for and cooperate in the support necessary when a child is nursed at home in the terminal stages.


The role of adherent cells in the secondary cell-mediated response in vitro to a natural poxvirus pathogen. The role of adherent cells in an in vitro secondary response to ectromelia virus infection was investigated. Spleen cells from ectromelia-primed mice ("responder" cells) depleted of adherent cells by either carbonyl iron treatment, adherence to plastic or passage through cotton wool columns had a markedly decreased capacity to produce a secondary response, as indicated by decreased T cell-mediated cytotoxicity against virus-infected target cells, when cultured with virus-infected "stimulator" cells. The secondary response was restored by the addition of peritoneal cells from either normal or ectromelia-immune mice. Small numbers of peritoneal cells completely reconstituted the response within a certain dose range but larger numbers produced a marked inhibition of the response. Spleen cells were less effective in restoring the response. The peritoneal cells were not merely acting as additional, infected "stimulator" or antigen-presenting cells, since they could be added as late as 3 days after culture. Reconstituting activity was not affected by pretreatment with anti-theta serum and complement and cell separation studies showed that the activity was associated mainly with Ig-negative cells and that the active cell probably bears Ia antigens on its surface. These results indicate that the adherent cells involved are probably macrophages and that they act non-specifically to produce optimum conditions for the specific response of T cells.


Interrelationships of copper, cytochrome oxidase, phospholipid synthesis and adenine nucleotide binding. Studies are reported on the interrelationships in liver mitochondria of copper status, cytochrome oxidase activity, adenine nucleotide binding capacity and phospholipid synthesis. Direct exposure of mitochondria to cyanide or diethyldithiocarbamate depressed cytochrome oxidase activity; ADP-binding and phospholipid synthesis. Fractionation of mitochondria to increase the specific activity of cytochrome oxidase about 10-fold did not increase the affinity to bind ADP. Ageing of mitochondria or dialysis of mitochondria or mitochondrial membrane preparations against water or diethyldithiocarbamate at 0--2 degrees for 18 h did not decrease cytochrome oxidase activity or copper content of reisolated and resuspended mitochondria or mitochondrial membrane preparations, but considerably reduced the affinity to bind ADP. The respiratory inhibitors, fluoride and azide, at concentrations inhibitory to cytochrome oxidase did not reduce ADP-binding or phospholipid synthesis. Atractyloside did not inhibit cytochrome oxidase activity but did inhibit ADP-binding and phospholipid synthesis. Pre-incubation of mitochondrial membrane preparations with Cu++ increased the copper content and ADP-binding affinity. The results indicate that cytochrome oxidase is not the ADP-binding site of the mitochondrial membrane system and that reduced cytochrome oxidase activity per se does not depress binding affinity. Copper appears to be a component of the adenine nucleotide binding sites of mitochondrial membranes because the copper-complexing agents, cyanide and diethyldithiocarbamate, depressed ADP-binding, while increased mitochondrial membrane copper content increased ADP-binding.


Watery diarrhoea syndrome with episodic hypercalcaemia. A patient with the Watery-Diarrhoea syndrome and episodic hypercalcaemia is reported. Plasma levels of vasoactive intestinal peptide (VIP) were elevated, and an islet cell adenoma of the pancreas was removed following which VIP levels decreased and diarrhoea ceased. During a hypercalcaemic episode, serum parathyroid hormone (PTh) levels were suppressed indicating the hypercalcaemia was independent of PTh and probably due to a direct action of VIP on calcium turnover.


Congenital cerebellar hypoplasia in jersey calves. Congenital cerebellar hypoplasia and hydrocephalus is reported in 8 Jersey calves. The possible relationship between this entity and BVDMD virus is discussed.


A fluorescent-antibody study of the pathogenesis of infectious laryngotracheitis. A study of the pathogenesis of infectious laryngotracheitis (ILT) by the fluorescent-antibody (FA) technique in birds inoculated ocularly, intratracheally, by vent brush, and by aerosol revealed that the epithelium of the conjunctiva and respiratory tract are the tissues actively infected by natural routes of infection. The virus propagated at the sites of initial exposure but there was no evidence of spread by viremia. Viral antigen was detected 2-7 days postexposure and rarely thereafter. Birds infected intratracheally showed intense infection in the nasal turbinates, suggesting that high concentrations of virus were expelled as an aerosol from infected tracheas. Virus applied to the dorsal lip of the vent by drop or brush was rapidly transferred mechanically to the bursa of Fabricius; the epithelial lining between follicles and between follicular cortex and medulla was positive in FA tests 48-72 hours postexposure.


Influence of environment on airsacculitis: effects of relative humidity and air temperature on broilers infected with Mycoplasma synoviae and infectious bronchitis. Mycoplasma synoviae (MS) obtained from broiler chickens condemned for airsacculitis was used to determine the influence of air temperature and relative humidity on the severity of airsacculitis produced experimentally. Infectious bronchitis virus was administered to 3-week-old broilers 5 days before aerosol exposure to MS broth cultures, producing extensive airsacculitis within 21-day study periods. High (31-32 C), medium (19-24 C), and low (7-10 C) air temperatures were studied in conjection with high (75-90%), medium (38-56%), and low (23-26%) relative humidities. Airsacculitis was most extensive (45%) at low temperatures regradless of high or medium humidity. The incidence of airsacculitis was greater (39%) at low humidity than at high humidity (17%) when air temperatures were medium. At high temperature, the trend was toward more airsacculitis (12%) at high humidity than (5%) at low humidity. However, the effect of cold air temperature was more dominant than the effect of relative humidity.


Pacheco's parrot disease of psittacine birds. Pacheco's parrot disease was identified as the cause of death of 3 psittacine birds at an aviary. Confirming a previous report, a herpesvirus was found to be the etiologic agent. The virus induced mortality in embryonated chicken eggs and budgerigars. The virus was characterized by electron microscopy, filtration experiments, and ether-sensitivity. Hepatic lesions in chick embryos and budgerigars were similar by light- and electron-microscope studies. Naked and enveloped nucleocapsids were seen in the nucleus and cytoplasm of infected hepatocytes, and naked virions were present in the sinusoids of the liver. Multinucleated cells in the liver contained virus in both intranuclear and intracytoplasmic inclusions; the latter inclusions were both membrane-bound and not. Vaccination of budgerigars with a commercial lyophilized Marek's disease vaccine did not prevent infection with the Pacheco's disease herpesvirus.


Virus-neutralizing antibody titers against 8 avian adenovirus serotypes in breeder hens in Georgia by a microneutralization procedure. Serums from 16 chicken-breeder flocks in Georgia were tested for virus-neutralizing antibody to 8 serotypes of avian adenovirus. Titers to all 8 serotypes were demonstrated in 8 of the flocks, titers to 7 in 6, and titers to fewer than 7 in the other 2 flocks. Although titers were high overall to some serotypes (types 2 and 8) and low to others (types 1, 4, and 5), with statistically significant differences between many titers, the data were difficult to interpret because of possible heterotypic responses of chickens infected with avian adeno-viruses. Used for titrations of serum antibody was a microneutralization procedure that is inexpensive. It proved also to provide reproducibility since titers from replicate tests differed by less than 3 twofold dilutions. In addition, there was a strong linear relation between titers obtained with the microneutralization procedure and a conventional plaque-assay titration procedure. The microneutralization procedure was less sensitive than the conventional procedure by a factor of about 5.5, but that was considered a disadvantage only with low-titered serums.


The pathology of necrotic enteritis of chickens following infusion of broth cultures of Clostridium perfringens into the duodenum. Necrotic enteritis was consistently reproduced when enough active broth culture of Clostridium perfringens type A was infused intraduodenally. Typical lesions of necrotic enteritis were seen as early as 5 hr after infusion was begun. The histologic lesions observed at 1 hr were characterized by edema in the lamina propria and desquamation of epithelial cells. Large numbers of clostridia were seen among these sloughed cells. Coagulation necrosis of the tips of villi became evident at 3 hr and was marked at 5 hr. Many clumps of clostridia were obvious among the necrotic tissue. At 8 and 12 hr the necrotic lesions extended to involve most of the villus structures. Morphologically abnormal erythrocytes were evident in the visceral organs at 12 hr.


The pathology of necrotic enteritis of chickens following infusion of crude toxins of Clostridium perfringens into the duodenum. Necrotic enteritis was produced in 4-week-old chickens with bacteria-free crude toxins of Clostridium perfringens. Typical gross lesions could be seen as early as 3 hr after intraduodenal infusion of toxin was begun. Early microscopic lesions were observed at 20 minutes, progressing with time to involve most of the mucosa. Death was related to the amount of toxin administered. Because systemic absorption of toxin is likely, acutely affected birds may die from toxemia.


The interaction of Clostridium perfringens and its toxins in the production of necrotic enteritis of chickens. The intraduodenal administration of large numbers of Clostridium perfringens cells harvested from broth cultures and resuspended in PBS or fresh sterile thioglycollate broth produced a very mild form of necrotic enteritis. Administering an appropriate number of cells in culture supernatant, however, produced typical field-type disease. Alpha toxin was shown to be the significant toxin recoverable from broth-culture supernatant fluids. Requirements to produce the disease are minor intestinal damage and sufficient numbers of toxigenic C. perfringens in the intestine.


Preparation of antisera to group-specific antigens of avian leukosis-sarcoma viruses: an alternate approach. Immunization of rabbits with a pooled preparation of chromato-graphically purified avian myeloblastosis virus (AMV) group-specific (gs) antigens produced relatively large volumes of antiserum that was as broadly reactive in complement-fixation (CF) tests as antiserum produced in hamsters with tumors induced by Rous sarcoma virus. This alternate procedure should be of value for routine preparation of leukosis virus gs antiserum. Other antisera prepared against disrupted AMV had spurious reactivities that confused CF testing of embryos and tissue extracts. This artifact was attributed to the high affinity of serum albumin for AMV virions and the resultant production of antibodies when plasma-derived AMV was used as immunogen. A high-molecular-weight AMV CF antigen was also found in early fractions eluted from an agarose-gel column.


Adenylate cyclase activity in lymphocyte subcellular fractions. Characterization of non-nuclear adenylate cyclase. Human peripheral lymphocytes were broken in a Dounce homogenizer and subcellular fractions enriched in plasma membranes or microsomal particles and mitochondria were isolated by centrifugation through a discontinuous sucrose gradient. Various agents that promote cyclic AMP accumulation in intact lymphocytes were compared in their ability to stimulate adenylate cyclase activity in the individual fractions. Plasma-membrane-rich fractions that were essentially free of other subcellular particles as judged by electron microscopy and marker enzyme measurements responded to fluoride, but weakly or not at all to prostaglandin E1 and other prostaglandins. Microsomal and mitochondrial-rich fractions responded markedly to both prostaglandin E1 and fluoride. In some, but not all, experiments phytohaemagglutinin produced a modest increase in enzyme activity in plasma-membrane-rich fractions. Catecholamines, histamine, parathyrin, glucagon and corticotropin produced little or no response. In the absence of theophylline, adenosine (1-10 micronM) stimulated basal enzyme activity, although at higher concentrations the responses to prostaglandin E1 and fluoride were inhibited. GTP (1-100 micronM) and GMP(5-1000 micronM) respectively inhibited or stimulated the response to fluoride, whereas the converse was true with prostaglandin E1.


Use of polyethylene glycol and fluorocarbon for the purification of avian encephalomyelitis virus. A simple method is described for isolating avian encephalomyelitis virus (AEV) from a bulk amount of proteins and lipids in a homogenate of infected whole chicken embryos. More than 90% of the virus in the homogenate was precipitated by 4% polyethylene glycol (PEG), and the precipitate was free of approximately 90% of the total nonviral protein. When the homogenate was extracted with fluorocarbon, the aqueous fraction retained the full virus infectivity and about 70% of the protein but was free from most lipid materials. A two-step procedure (precipitation of the homogenate by 4% PEG, followed by extraction of the precipitate by fluorocarbon) gave a 50-fold purification of AEV over the original homogenate. The procedure is simple, rapid, and effective in removing nonviral proteins and lipids from host tissues. It is useful as an initial treatment of embryonic materials for the purification of AEV.


Adenylate cyclase activity in lymphocyte subcellular fractions. Characterization of a nuclear adenylate cyclase. Nuclei from purified human peripheral lymphocytes were prepared by incubations with Triton X-100 to disrupt the cells, followed by sucrose-density gradient centrifugation. The nuclei were pure as judged by phase-contrast microscopy and had low contents of non-nuclear marker enzymes. In addition, nuclei prepared from lymphocytes surface-labelled with 125I had only 2-7% of the radioactivity bound to intact lymphocytes. At 3.3 mM-Ca2+ and 100 micronM-ATP a fluoride-sensitive adenylate cyclase was demonstrated in nuclei prepared in 0.2% Triton X-100 or 0.33% Triton X-100. There was linear accumulation of cyclic AMP for 10 min in both preparations. The apparent Km for ATP was 90 micronM. Adenylate cyclase activity was augmented by 1.0 mM-Mn2+ and inhibited at higher concentrations. Ca2+ showed two peaks of stimulation, at 1.0-2.5 mM- and above 10 mM-Ca2+. Mg2+ was inhibitory at all concentrations. EDTA OR EGTA only slightly decreased adenylate cyclase activity, suggesting that another metal ion may be necessary for activity. Adenylate cyclase activity was stimulated by 10mM-isoproterenol and 10 micronM-adrenaline in the presence of a phosphodiesterase inhibitor. Phytohaemagglutinin and prostaglandin E1 alone or in combination with isoproterenol had no effect on nuclear adenylate cyclase activity in either nuclei preparation. These results indicate that human lymphocyte nuclei contain one or several adenylate cyclases which differ from adenylate cyclases found in other subcellular fractions of these cells with regard to their bivalentcation requirements and responsiveness to pharmacological agents.


Effects of adenosine 3':5'-cyclic monophosphate and serum on synthesis of hyaluronic acid in confluent rat fibroblasts. A small amount of hyaluronic acid is synthesized in confluent cultures of rat fibroblasts, which have a high content of cyclic AMP. Addition of calf serum caused a rapid decrease in the cellular cyclic AMP content and large increases in hyaluronic acid synthetase activity and hyaluronic acid production. Addition of cyclic AMP also caused a marked increase in hyaluronic acid synthetase activity within 2h and then increased hyaluronic acid production. The effects of cyclic AMP and serum on hyaluronic acid synthesis were additive. Prostaglandin E2, which increased the cyclic AMP by stimulating adenylate cyclase, was as effective as cyclic AMP in increasing hyaluronic acid synthetase activity, but AMP was far less effective than cyclic AMP. These results indicate that cyclic AMP itself stimulates the mucopolysaccharide synthesis and that the effect of serum is not due to a decrease in cyclic AMP in the cells.


Effects of glucose, glucose metabolites and calcium ions on adenylate cyclase activity in homogenates of mouse pancreatic islets. The effects of glucose, a series of glucose metabolites, nicotinamide nucleotides, Ca2+ and p-chloromercuribenzenesulphonate on adenylate cyclase activity in homogenates of mouse pancreatic islets were studied. The basal activity of the adenylate cyclase was approx. 6 pmol of cyclic AMP formed/30 min per microng of DNA at 30 degrees C. The enzyme activity was stimulated by some 150% by fluoride. Starvation of the animals for 48h had no effect on either the basal or the fluoride-stimulated activity. The adenylate cyclase activity was increased by 40-50% when 17 mM-glucose, 10 micronM-phosphoenolpyruvate or 10 micronM-pyruvate was added to the assay medium. The effect of glucose was unchanged in the presence of 17 mM-mannoheptulose, and mannoheptulose alone had no effect. The other glycolytic intermediates, and the coenzymes NAD+, NADH and NADPH, at concentrations up to 1 mM were without any detectable effect on the rate of formation of cyclic AMP. The insulin secretagogue p-chloromercuribenzenesulphonate inhibited the adenylate cyclase markedly even at a concentration of 10 micronM. Calculated concentrations of free Ca2+ of 10 micronM and 0.1 mM inhibited adenylate cyclase by 29 and 71% respectively. It is concluded that both glucose itself and phosphoenolpyruvate and/or pyruvate are true activating ligands for islet and adenylate cyclase and that inhibition of the cyclase by Ca2+ may be of physiological significance.


The appearance of gluconeogenesis at birth in sheep. Activation of the pathway associated with blood oxygenation. 1. Measurements of pyruvate carboxylase, mitochondrial phosphoenolpyruvate carboxykinase (GTP), hexose bisphosphatase and glucose 6-phosphatase in developing sheep liver showed substantial activities of all enzymes in the foetus, especially towards the end of gestation. Cytosol phosphoenolpyruvate carboxykinase (GTP) in livers of mid-term foetuses was only 10% of the activity at birth. 2. All enzymes except pyruvate carboxylase showed 1.5-2-fold increases after birth. 3. Gluconeogenesis form [14C]actate could not be detected in chronically cannulated sheep foetuses at any developmental stage and was not initiated by the infusion of adrenaline or glucagon. 4. An active pathway of gluconeogenesis was evident in vivo within 2 min after natural birth or within 4 min after Caesarian delivery of term lambs, and was delayed in prematurely delivered lambs until breathing was established and the blood fully oxygenated. 5. It is proposed that oxygen availability initiates gluconeogenesis in the newborn lamb.


Some properties of aldehyde dehydrogenase from sheep liver mitochondria. Aldehyde dehydrogenase from sheep liver mitochondria was purified to homogeneity as judged by electrophoresis on polyacrylamide gels, and by sedimentation-equilibrium experiments in the analytical ultracentrifuge. The enzyme has a molecular weight of 198000 and a subunit size of 48000, indicating that the molecule is a tetramer. Fluorescence and spectrophotometric titrations indicate that each subunit can bind 1 molecule of NADH. Enzymic activity is completely blocked by reaction of 4mol of 5,5'-dithiobis-(2-nitrobenzoate)/mol of enzyme. Excess of disulfiram or iodoacetamide decreases activity to only 50% of the control value, and only two thiol groups per molecule are apparently modified by these reagents.


The allosteric mechanism of bovine liver glutamate dehydrogenase. Evidence from circular-dichroism studies for a conformational change in the ternary complex enzyme-(oxidized nicotinamide-adenine dinucleotide)-glutarate. 1. Computer averaging of multiple scans was used to refine the circular dichroism spectrum of bovine liver glutamate dehydrogenase, revealing well-defined structure in the aromatic region. 2. The circular dichroism of NAD+ bound to glutamate dehydrogenase is strongly negative at 260nm, probably owing to immobilization of the adenosine moiety. Loss of the characteristic adenine-nicotinamide interaction suggests that the coenzyme is bound in an unstacked conformation. 3. Glutarate and succinate, substrate analogues that are both inhibitors competitive with glutamate, do not significantly perturb the circular-dichroism spectrum of the enzyme in the absence of NAD+. 4. In the presence of NAD+, 150nM-succinate decreases the negative circular dichroism corresponding to bound coenzyme, but does not affect the protein circular dichroism. However, ISOmM-glutarate causes profound alternations of the circular-dichroism spectra of the bound NAD+ and of the enzyme, indicative of a protein conformational change. This direct evidence of conformational change specifically promoted by C5 dicarboxylates confirms the previous inference from protection studies. 5. The conformational change is discussed in relation to the allosteric mechanism of glutamate dehydrogenase.


Evidence that latent collagenases are enzyme-inhibitor complexes. Specific collagenase from the culture media of various rabbit tissues and cells exists in active and latent forms. Latent collagenase is most effectively activated with 4-aminophenylmercuric acetate, a thiol-blocking reagent, strongly suggesting that latent forms are enzyme-inhibitor complexes. A collagenase inhibitor from bone cultures, which may be closely related to the inhibitor of such latent enzyme complexes, was partially characterized.


The amino acid sequence of cytochrome c from the locust, Schistocerca gregaria Forskal. The amino acid sequence of locust cytochrome c was determined, although the overlap between chymotryptic and tryptic peptides at residues tyrosine-97 and leucine-98 was not observed, owing to an anomalous tryptic break duplicating the chymotryptic digestion. The molecule consists of a single polypeptide chain of 107 residues, homologous with other mitochondrial cytochromes c. In common with other known insect cytochromes c, it possesses a non-acetylated, four-residue tail at the N-terminus relative to glycine-1 of the standard alignment. A molecular phylogeny for 17 species was constructed relating the cytochrome c molecules of Schistocerca gregaria and other invertebrates with those of representative taxonomic groups. Experimental details are given in a supplementary paper deposited as Supplementary Publication SUP 50077 (24 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can obtained on the terms indicated in Biochem. J. (1977) 161, 1.


The influence of the calcium antagonist fendiline on tone and motility of the guinea pig gut smooth muscle and the cAMP and cGMP concentrations of the isolated terminal ileum. In various parts of the guinea pig gastrointestinal tract the calcium antagonist N-(2-benzhydryl-ethyl)-N-(1-phenyl-ethyl)-amine hydrochloride (fendiline, Sensit) decreases the smooth muscle tone elevated by K+-induced depolarisation. This effect is antagonized by addition of extra-Ca++. The muscle relaxation is dos-dependent and amounts to 45-90% after 1-5 microng/ml fendiline. Proportionally to this effect the tissue concentration in cGMP is decreased whereas cAMP remains unchanged. After 54 micron/ml theophylline the cAMP level in the terminal ileum is increased significantly whereas cGMP does not change. Theophylline has no influence on the relaxing effect of 1 microng/ml fendiline. By contrast, the increase in cAMP after theophylline is prevented by fendiline. These findings are explained by the antagonistic effect of fendiline to Ca++, which activates the guanylate cyclase and inhibits the adenylate cyclase. Furthermore, fendiline seems to prevent the binding of theophylline to guinea pig ileal phosphodiesterase. It is discussed that cGMP plays a physiological role in controlling the intestinal smolth muscle tone and motility.


Changes in muscle action potentials of patients with diseases of motor units following the infusion of a peptide fragment of ACTH. The polypeptides ACTH and ATCH4-10 (OI 63) witha sequence of amino acids H-Met-Glu-His-Phe-Arg-Trp-Gly-OH, have similar stimulating effects  on motor units in lower mammals. Their actions differ primarily in that ACTH4-10 is not corticotropic. Since the corticotropic action of ACTH frequently presents a problem during its clinical use in treatment of motor unit diseases, the action of ACTH4-10 was studied in two patients with muscular atrophy. Prior to administration of ACTH4-10, stimulation of M. oppenens pollicis through the median nerve evoked muscle action potentials in both patients which progressively declined in amplitude. This decline was not observed subsequent to the infuscion of ACTH4-10 (3, 6 and 15 mg). The effect lasted partially in excess of at least 2 h. It is suggested that ACTH4-10 produces this effect by direct action on a peripheral component of the motor unit and/or indirectly by an action on the central nervous system.


Effect of vinblastine on rat liver ultrastructure. A single injection of vinblastine given i.v. to rats lead to the disappearance of microtubules, development of crystalline inclusions, dilation of Golgi sacculi, formation of multivesicular bodies and cytosegresomes as well as accumulation of lysosomes in the cytoplasm of liver cells. The endoplasmic reticulum and mitochondria do not seem to be affected. The question of whether or not these fine structural alterations are causally related has yet to be elucidated.


[The immune response of the fetus in the diagnosis of intrauterine infections: IgM and IgA concentrations in the umbilical cord blood and titers of rubella and cytomegalic virus antibodies]. Two serological methods, hemagglutination and complement fixation, have been used respectively to determine HAI titers to rubella virus and C fixing antibodies to cytomegalovirus in cord blood of newborn infants delivered in Turin from August 1973 to April 1974. Sera have been divided according to antibody titer and aspecific IgM and IgA concentrations. By statistical analysis we observed a highly significant association between both IgM and IgA concentration and antibody titers to rubella virus.


Neuromuscular facilitation during train-of-four and tetanic stimulation in healthy volunteers: observations with half-refractory paired responses. The compound electromyographic response of the thenar musculature was elicited by supramaximal stimulation of the ulnar nerve in 12 awake healthy volunteers. The half-refractory period of the neuromuscular transmission was determined. Twin stimuli separated by the half-refractory interval were repeated as a unit at 2 Hz and at 50 Hz. Thus two trains of responses, one beginning with a maximal response, the other with a half-maximal response, were obtained with each frequently of stimulation. Marked facilitation of the electromyographic response began with the train of stimulation, especially with tetanic stimulation. A secondary fade occurred occasionally. These changes were more remarkable in the train of half-refractory responses. We conclude that nerve stimulation facilitates neuromuscular transmission and deduce that the characteristic fade observed with a curariform block, rather than being an uncovered physiological phenomenon, is caused by tubocurarine and related drugs.


C-type and intracisternal A-type virus particles during epidermal carcinogenesis by tobacco smoke condensate in BALB/c mice. Electron microscopic observations of sequential stages of skin carcinogenesis induced by tobacco smoke condensate (SC) and a cyclohexane fraction of tobacco smoke condensate (G) revealed an increase in incidence of intracisternal A particles within the epidermal cells. Tumours induced by SC also contained C-type particles, but these were not seen in G-induced tumours or after irritant or solvent treatment. There was no evidence of an increase in intracisternal A particles after irritant or solvent treatment. A direct relationship between the proliferation of A particles and neoplastic growth of BALB/c mouse epidermis appears likely. The data suggest possible activation of a latent C-type virus by SC.


Calcitonin-responsive adenylate cyclase in a calcitonin-producing human cancer cell line. A calcitonin-responsive adenylate cyclase has been found in a cell line of a poorly differentiated bronchial carcinoma (BEN cells). The cells have previously been shown to secrete an immunoreactive form of calcitonin in culture. Salmon calcitonin (SCT), porcine calcitonin (PCT) and human calcitonin (CT-M) all stimulated adenylate cyclase activity in particulate preparations. CT-M sulphoxide had little effect. The concentrations of the calcitonins required for half the maximum activation of adenylate cyclase were 6-8, 18 and 90 nm respectively. SCT (30pm) and CT-M (60 pm) increased the intracellular concentration of cyclic AMP from 11-2+/-0-2 (s.e.) to 18-2+/-0-2 and 16-7+/-0-2 respectively over a 2-5-min period. SCT (labelled with 125I) bound to particulate preparations of Ben cells, and competition for binding occurred with unlabelled SCT and CT-M. The concentration of SCT required for half the maximum inhibition of [125I]SCT binding was 11 nm. CT-M sulphoxide inhibited only at high concentration (3 micron). The characteristics of the adenylate cyclase response to SCT did not change over the period between cell adhesion (after subculture) and confluence. However, pre-incubation of cells for 4 h with SCT (150 nm) abolished the subsequent adenylate cyclase response of particulate preparations to further hormone. The practical difficulties encountered in purifying and quantifying the large-mol.-wt. form of CT-M secreted by BEN cells has precluded direct investigation of the potential relationship between hormone secretion and the occurrence of the calcitonin receptor. This relationship is discussed in terms of its possible biological significance.


Properties of a cell line from human adenocarcinoma of the rectum. A new, highly differentiated line of cells derived from adenocarcinoma of the rectum (HT55) is described. This line is noteworthy for the following features: 1. The role played in its development by the use of UV-inactivated Sendai virus to attach tumour cell clumps to plastic bottles. 2. Evidence that it produces RNA-containing material of density 1-5--1-16 g/ml. 3. Induction of bone formation in the stroma when grown in athymic mice. 4. Stimulation of primary CBA mouse embryo fibroblasts to form a transient nodule when mixed with them and injected into adult CBA mice. The karyotype and growth-cycle characteristics of the line are described.


Evaluation of 51Cr release for detecting cell-mediated cytotoxic responses to solid chemically induced rat tumours. A 51Cr-release test was developed for the detection of cell-mediated cytotoxicity against transplanted solid chemically induced rat tumours, and the findings were compared with those obtained in parallel tests using a microcytotoxicity assay. It was necessary to incorporate several modifications of the original Brunner assay (Brunner et al., 1968) in order to increase the sensitivity of the test as applied to long-term tumour lines maintained as glass-adherent cultures. These included: 1. preincubation of labelled tumour cells at 37 degrees C for 3-4 h before addition of effector cells; 2. preincubation of effector cells at 37 degrees C for 12 h before addition to target cells; 3. upon admixture of target and effector cells, intimate cell contact was established by gentle centrifugation; 4. after incubation of target and effector cells at 37 degrees C, a further incubation for 1 h at 45 degrees C was included to complete the release of 51Cr from non-viable cells. Maximal cytotoxicity of tumour-bearer effector cells was detected after 12 h incubation of lymphoid and target cells at a ratio of 200:1. Spleen cells from tumour-bearing rats during the first 2 weeks of tumour growth exhibited the same pattern of reactivity in the 51Cr-release test and the microcytotoxicity assay, although significant reduction in target-cell numbers was more frequently recorded using the microcytotoxicity assay. Tumour-bearer spleen cells showed reactivity against the homologous tumour as well as against unrelated tumours. Using either assay, pre-exposure of effector cells to 3M KC1 extracts of tumour was found to inhibit their effect on tumour cells most frequently in tests in which the effector-cell donor, soluble extract and target cell were of the same tumour line.


Abnormal platelet function in Chediak-Higashi syndrome. Platelets in an infant with Chediak-Higashi (C-H) syndrome without bleeding manifestations and not in the accelerated phase showed abnormal function consistent with storage pool disorder as shown by abnormal aggregation, decreased storage capacity and release of [14C]5-HT, low endogenous 5-HT, reduced ATP and ADP with an increased ATP/ADP ratio, increased specific radioactivity of ADP after [14C]adenine labelling, decreased release of adenine nucleotides after stimulation, impaired secretion of acid hydrolases despite normal stores, and decreased calcium content. Incorporation of [14C]adenine into metabolic pool adenine nucleotides was normal. Nucleotide conversion to hypoxanthine in stimulated platelets was mildly impaired. Platelet cyclic-AMP (c-AMP) was initially elevated, but even when c-AMP returned to normal levels after ascorbate treatment, platelet function was not improved. Elevated intracellular c-AMP was not solely responsible for the abnormal platelet function.


Simultaneous presence of receptors for complement and sheep red blood cells on human fetal thymocytes. The cells of four thymus glands, two at 12 weeks, one at 16 weeks and one at 22 weeks gestational age, were examined cytochemically for enzymic activity and for surface markers. The reactions for chloroacetate esterase, peroxidase, and alkaline phosphatase were negative and the reaction for nonspecific esterase was only weakly positive in some of the cells. In contrast, nearly all thymocytes at the 12th gestational week showed strong focal acid phosphatase (acP) activity. The number of acP-stainable cells and the staining intensity declined progressively. A high percentage of thymocytes in early fetal life express a complement receptor (CR); the proportion of CR-bearing cells decreased while cells forming rosettes with sheep erythrocytes (possessing erythrocyte receptors: ER) correspondingly increased during fetal life. Using a mixed rosette assay, up to 30% of the thymocytes at 12 weeks gestation were found to bear CR and ER simultaneously. The number of CR- and ER-positive cells also declined progressively with fetal age. These findings show that CR-positive, ER-negative thymus cells mature into CR-negative, ER-positive thymocytes via a CR-positive and ER-positive intermediate cell, indicating that mature ER-positive thymocytes do not originate from another cell line lacking CR. The changes in surface markers during early T cell maturation is relevant to certain lymphomas whose cells also show strong focal acP activity and also express both CR and ER. These lymphoma cells may correspond to fetal thymocytes or T precursor cells present in small numbers after birth.


Effect of hyperoxia on superoxide anion and hydrogen peroxide production of polymorphonuclear leucocytes and alveolar macrophages. Hyperoxia activates superoxide dismutase (SOD) while inactivating catalase and glutathione peroxidase in polymorphonuclear leucocytes (PMN) and alveolar marcophages (AM) obtained from guinea-pigs exposed to 85% oxygen for 90 h. The influence of these altered enzyme activities on the rate of oxygen consumption and release of superoxide anion (O--2) and hydrogen peroxide (H2O2) was investigated. By 18 h O--2 released from resting PMN increased two-fold and remained elevated through the entire periods of the study, whereas H2O2 release and oxygen consumption at the same time points remained normal. At 66 h PMN phagocytizing opsonized zymosan particles released additional quantities of O--2 and H2O2 and consumed significantly more oxygen compared to the usual increase noted at earlier time points. Although oxygen consumption was almost two-fold higher in AM than PMN, phagocytizing AM released three-fold less O--2 and five-fold less H2O2 than did PMN. Furthermore, AM of animals exposed to hyperoxia no longer exhibited enhanced O--2 production upon exposure to opsonized zymosan. Hydrogen peroxide release progressively decreased at rest but progressively increased during phagocytosis of opsonized zymosan during the 90 h exposure to hyperoxia. No changes in oxygen consumption of AM occurred during hyperoxia. The divergent oxidative responses in PMN and AM of guinea-pigs exposed to hyperoxia suggest different biochemical adaptive mechanisms.


[Phospholipid composition of nuclear membranes and cell nuclei of rat liver and hepatoma 27]. Phospholipid composition of nuclei and nuclear membranes from rat liver and hepatoma-27 were investigated. Hepatoma nuclei and nuclear membranes were found to contain cardiolipin which was absent in the same fractions of rat liver. In the nuclei and nuclear membranes of hepatoma the content of sphingomyelin was higher and that of lecithin is lower than in the corresponding subcellular fractions of  rat liver. The content of acid phospholipids was much higher in hepatoma nuclear membranes than those of the normal liver. The data obtained show that the general trend of lipid dedifferentiation which earlier was demonstrated for various membranes of different tumor cells is observed also in the case of nuclear membranes.


[Use of specific angiotensin inhibitors in the study of hormone-receptor interaction]. The specificity of receptors participating in the interaction with angiotensin II (AT 1--8) fragments, e. g. N-terminal tri-(AT 1--3), C-terminal penta-(AT 4--8), and middle tetrapeptide (at 3--6), was studied in experiments on rat ascending colon which were performed to investigate the influence of a specific angiotensin antagonist, /1-hydantoic acid, 5-valine, 8-alanine/-angiotensin (HAAT 1--8), on myotropic effects of these fragments as well as the influence of the fragments on the myotropic activity of the whole hormone molecule. Competitive antagonism was found between HAAT 1--8 and AT 4--8 and AT 3--6 and between these fragments and AT 1--8. No antagonisma was revealed between AT 1--3 and HAAT 1--8, and between AT 1--3 and AT 1--8. It is assumed that the fragments AT 4--8 and AT 3--6 interact at the level of angiotensin receptors, and that of AT 1--3 at the level of non-specific receptors. A new model for the structural and functional organization of angiotensin is proposed.


[Electrogenic function of submitochondrial particles at the water-octane interphases]. Studies on submitochondrial particles (SMP) preparation showed that in the sourse of the redox reactions at the octane-water interface, catalyzed by SMP enzymes, the charges are transferred from the aqueous to the octane phase. The effects were detected by a shift of the Volta potential, using the vibrating electrode method. In the presence of 2-N-methyl-amino-1,4-naphthoquinone in octane, acting as electron acceptor, the negative charges were transferred from water to octane following the oxidation of NADH, succinate and ascorbate. The charging of the octane phase was sensitive to the inhibitors of the respiratory chain, e. g. rotenone, antimycin and cyanide. In the presence of 2,4-DNP in octane, acting as a proton acceptor, the oxidation of NADH and succinate by ferricyanide, catalyzed by CMP in the presence of antimycin and cyanide correspondingly, was followed by a transfer of positive charges from water to octane. The positive charging of the octane phase, coupled with NADH oxidation, was found insensitive to rotenone, and that coupled with succinate oxidation, was completely inhibited by antimycin. The positive charging of the octane phase was also observed during the reverse transhydrogenase reaction, catalyzed by SMP at the division of the phases. The effect was inhibited by palmitoyl-CoA.


[Detection of multiple forms of polyphosphate phosphohydrolase from Endomyces magnusii]. Labile polyphosphate phosphohydrolase from Endomyces magnusii is 27-fold purified by means of fractionation with ammonium sulphate, gel filtration on Sephadex G-75 and Biogel P-60 and chromatography on DEAE cellulose. Chromatography on DEAE Sephadex A-50, isoelelctric focusing and polyacrylamide gel electrophoresis of the enzyme preparation revealed 3 different fractions with polyphosphate phosphohydrolase activity (PPPH1, PPPH2 and PPPH3). Relative content of these fractions in E. magnusii cells is 30%, 55% and 15% respectively. Isoelectric points are: PPPH1--pH 5.1--5.2; PPPH2--pH 6.0--6.1; PPPH3--pH 6.3--6.4. PPPH1 and PPPH2 are found to be the most labile. PPPH3 is more stable under isolation procedure and storage. The fractions have similar molecular weight (48 000 +/- 3000).


[Sulfhydryl groups of cyclic AMP-dependent histone kinase from pig brain]. A preparative method of isolation of the histone kinase regulatory subunit has been developed. The total number of cysteine residues in the enzyme (13 residues) and in each of its subunits (3 residues in the catalytic subunit and 10 residues in the regulatory one), has been determined. Two disulfide bridges have been found in the regulatory subunits. The relative availability of the sulfhydryl groups with respect to different modifying agents has been studied. It has been demonstrated that SH-groups blocking does not change essentially the enzyme activity.


[Structural-functional changes in the synaptic membranes of the cerebral cortex of rats during electric stimulation in vitro]. Electric stimulation (EC) of a suspension of native synaptic membranes of rat brain cortex in the Krebs-Ringer-glucose medium revealed Ca-dependent inhibition of Na+, K+-ATPase and inhibition of transport Ca-activated, Mg-dependent ATPase. The effects observed are not induced by a change in the SH-groups of the membrane proteins and are removed by an addition of total lipids of the brain (membrane protein: lipid = 5:1) or 0.35 mM novocaine. Cyclic 3',5'-AMP in concentrations of 0.1--1.0 mM causes an inhibition (up to 50%) of Na+, K+-ATPase of native synaptic membranes. The Na+, K+-ATPase activity of purified membrane preparations is not changed either by the cyclic nucleotide, or by EC. It is assumed that depolarization of excitable membranes results in structural changes, mediated by the activation of protein kinase, and manifesting themselves as labilization of protein-lipid ratios.


[Various enzymes of isolated nuclear membranes and cell nuclei of the liver and hepatoma 27 of rats]. A comparative study of glucose-6-phosphatase, alcaline RNase, ATPase, inosine diphosphatase and 5'-nucleotidase activities in isolated rat liver and hepatoma-27 nuclei and nuclear envelopes was performed. The tumor nuclear membranes were shown to be free from G-6-Pase activity in contrast to the liver nuclear membranes. The nuclear RNase activity was strongly inhibited in the hepatoma and could be unmasked in the presence of 3-10(-4) M pCMB. Hepatoma nuclear and nuclear envelopes ATP-ase activity was found to be moderately decreased as compared to those of the normal tissue. The values of inosine diphosphatase activity in hepatoma were similar to those in liver. The role of the nuclear envelope in nuclear-cytoplasmic interactions as well as nuclear location of G-6-Pase are discussed.


Possible mechanisms involved in the stereotyped behavior elicited by amphetamine. In a variety of animals, amphetamine administration produces an increase in locomotor behavior and an induction of repetitive, stereotyped behaviors. There is now considerable evidence to suggest that the induction of stereotyped behaviors is accomplished, in part, by alterations in catecholaminergic transmission in the central nervous system. By recording the spontaneous activity of neurons in the rat brain substantia nigra, reticular formation, basal ganglia, and elsewhere during systemic administration of amphetamine and related drugs, or during administration by means of microinfusions directly into these brain regions, relationships may be drawn between the biochemical and behavioral effects of these drugs and drug-induced changes in neuronal activity in the central nerovous system. Current evidence, for example, suggests that amphetamine produces an inhibition of neuronal activity in the neostriatum and pars compacta of the substantia nigra by means of dopamine released from dopaminergic terminals in the neostriatum and dopaminergic dendrites in the substantia nigra respectively. In addition, current evidence suggests the possibility of a GABA-mediated functional antagonism between excitatory cortical and/or thalamic input to the neostriatum and dopaminergic input from the substantia nigra which could be involved in the apparently mutually exclusive occurrence of amphetamine-induced locomotion and stereotyped behaviors that follow amphetamine administration. Such evidence may also have relevance to a variety of behavioral disorders involving the basal ganglia and catecholaminergic transmission in the central nervous system.


The pathophysiologic basis of tardive dyskinesia. The prolonged course of tardive dyskinesia (TD) associated with antipsychotic drugs suggests that permanent structural alterations of the brain occur, though neurohistopathological studies have provided little to support this view. An alternative view is that functional adaptive changes may account for the manifestations of TD. A currently popular hypothesis is that there may be a functional excess in the activity of dopamine (DA) as a synaptic neurotransmitter in the basal ganglia. A cholinergic mechanism has also been implicated in TD, but the clinical effects of acetylcholine agonists and antagonists in TD are variable. In animals, nigrostriatal neurons respond to the blockade of DA synapses by treatment with antipsychotic agents in several ways, including acute and transient increases in the turnover of DA, and more slowly evolving "disuse" supersensitivity, possibly of postsynaptic receptors. The latter effects have been studied extensively in animal models of presumably DA-mediated behavior, by chemical studies of DA-sensitive adenylate cyclase in caudate nucleus in vitro, and by studies of labeled dopamine receptors. The phenomenon of DA-supersensitivity might help to explain some of the acute "withdrawal dyskinesias" that follow the abrupt discontinuation of high doses of antipsychotic agents, and might contribute to other reversible forms of the syndrome, but may be too short-lived to explain the persistent forms of TD. It must be concluded that an explanation of the latter syndrome of persistent drug-related TD remains uncertain.


Human cytomegalovirus: a review of developments between 1970 and 1976. Part II. Experimental developments. Over the last 6 years, our knowledge of the basic characteristics of human cytomegalovirus (HCMV) has been greatly enhanced, especially as concerns the microbiology of this virus. DNA relatedness studies show that there is 80% homology between different strains, regardless of their origin. Studies on the relationships between HCMV and host-cell metabolism reveal that it  induces both a virus specific and a cellular DNA polymerase, stimulates ribosomal RNA, induces early protein synthesis and results in the liberation of "intrinsic" interferon. The sequence of appearance of virus-specific antigens, as well as the presence of virus-induced IgG-Fc receptors has been described. New morphological aspects of the virus itself, as well as of the lesions which it provokes, have been observed. Last, but perhaps most important, evidence is accumulating in favor of the oncogenic potential of this virus. These basic aspects of HCMV are presented and discussed herein, along with the effects of various drugs and supra-optimal temperature on the virus and the possibility of preparing a vaccine against it.


Health of the intensively treated hemophiliac, with special reference to abnormal liver chemistries and splenomegaly. Liver function abnormalities have been noted in intensively treated hemophiliacs, and have led to less aggressive application of pooled plasma products by some physicians. In a prospective study, liver function was abnormal in 68 of 98 hemophiliacs. The abnormalities of hepatic function tended to persist over a 1-yr study period. There was no correlation between these abnormalities and the age of the patient, the presence of hepatitis-associated antigen or antibody, the presence or absence of splenomegaly (which was found in 26 of 98 patients), the number of infusions of plasma products, the type of hemophilia, or the type of product infused. Titers of antibodies to cytomegalovirus were generally higher in hemophilic patients than in a control group of healthy volunteers. These abnormalities did not suggest that a less aggressive infusion regimen was indicated for the hemophiliac, but did suggest the need for careful long-term observation of such patients.


Intracellular enzymes of collagen biosynthesis in human platelets. Activities of four intracellular enzymes of collagen biosynthesis--prolyl hydroxylase, lysyl hydroxylase, collagen galactosyltransferase, and collagen glucosyltransferase--were demonstrated in human platelets, and the presence of prolyl hydroxylase protein was further demonstrated by direct radioimmunoassay. The ratio of the specific activities of the four enzymes in the four enzymes in the human platelet extract to those in human adult skin extract varied from about 0.1 to 1, the lowest relative activity being found with prolyl hydroxylase and the highest with collagen glucosyltransferase. Only a very small amount of prolyl hydroxylase protein, probably 1%, was in the form of the active enzyme tetramer. The collagen glucosyltransferase from human platelets readily glucosylated galactosylhydroxylysine in denatured collagen, but did not glucosylate native collagen. Also, native collagen did not act as an inhibitor of the glucosylation reaction. Therefore, platelet collagen glucosyltransferase cannot form either an enzyme--substrate complex or an enzyme--inhibitor complex with native collagen. The results thus argue against the theory which maintains that platelet collagen glucosyltransferase is involved in collagen--platelet adhesion.


Parallel tubular arrays in severe combined immunodeficiency disease: an ultrastructural study of peripheral blood lymphocytes. The ultrastructure of the lymphocytes from three children with severe combined immunodeficiency disease (SCID) is described. Parallel tubular arrays (PTA) were found in a large percentage of circulating lymphocytes (53%, 41%, and 13%) in three SCID patients when compared to age-matched controls. The size of these inclusions was quite variable, with some attaining a length of 1.7 micrometer. They contained a tubular substructure with a diameter of 36--44 nm. The PTA were mostly located in the centriolar and Golgi regions of the cytoplasm, and were sometimes membrane bound. A centriolar origin of the inclusion was suggested. A second inclusion, the tubuloreticular structure, was found in only 1.4% of the circulating lymphocytes from one SCID patient. The origin of the PTA and its occurrence in severe combined immunodeficiency disease are discussed.


Myeloperoxidase--H2O2--halide system: cytotoxic effect on human blood leukocytes. Myeloperoxidase, H2O2, and a halide form a potent antimicrobial and cytotoxic system of the polymorphonuclear leukocyte. A cytotoxic effect of this system on human blood leukocytes is demonstrated, employing 51Cr release and dye exclusion assays. Cytotoxicity is dependent on enzymatically active myeloperoxidase, H2O2, or a peroxide-generating enzyme system and either chloride or iodide. Cell damage is rapid, with maximal levels of 51Cr release occurring within 30--60 min. Approximately equal sensitivity to the peroxidase system is observed for polymorphonuclear leukocytes and mononuclear leukocytes. Since myeloperoxidase and H2O2 are released from polymorphonuclear leukocytes under certain conditions, such as during particle ingestion, it is suggested that peroxidase-mediated leukocyte injury may be an important feature of the inflammatory response.


Metabolic activity of phagocytosing granulocytes in chronic granulocytic leukemia: ultrastructural observation of a degranulation defect. The functional capacities of granulocytes in patients with chronic granulocytic leukemia are still a subject of controversy, probably due to the heterogeneity of the abnormalities observed from patient to patient. For a better definition of these abnormalities, 14 patients with untreated chronic granulocytic leukemia were studied. The patients were divided into three groups on the basis of the functional activities of their phagocytosing granulocytes. In four patients (group I), the granulocytes were normal in respect to particle ingestion, nitroblue tetrazolium (NBT)-stimulated reduction, cyanide-insensitive oxygen (O2) consumption, superoxide anion (O2-)-stimulated production, hydrogen peroxide (H2O2) production, and iodination. They also had a normal myeloperoxidase (MPO) content. In four patients (group III), the granulocytes were significantly defective in all of these activities. In the six remaining patients (group II), all the initial metabolic steps of the phagocytosing granulocytes (ingestion, NBT reduction, O2 consumption, O2-production, H2O2 production) were normal, as were the MPO content of the granulocytes, while iodination was strikingly decreased. These metabolic features suggested a degranulation defect which was observed ultrastructurally in the only patient studied among these six. The phagocytosing granulocytes of this patient did not degranulate and no deposits of MPO activity were seen in the phagosomes.


Cold agglutinins in infectious mononucleosis and heterophil-antibody-negative mononucleosis-like syndromes. Cold agglutinins (CA) were evaluated prospectively in patients with various mononucleosis syndromes and in a large control group. Cold agglutinins with anti-i specificity were seen mainly in heterophil-positive or -negative Epstein-Barr virus (EBV)-induced infectious mononucleosis (31.8% of cases). Unclassified CA with equal reactivity against cord and adult erythrocytes were seen in 56 of 150 (37.3%) cases of heterophil-antibody-positive infectious mononucleosis (IM), in 1 of 7 (14.3%) cases of heterophil-negative EBV-induced IM, and in 12 of 31 (38.7%) cases of the heterophil-negative mononucleosis-like syndrome due to cytomegalovirus or other unspecified agents. One patient with heterophil-positive IM had a persistent, partially papain sensitive CA with anti-Pr-like activity. Anti-i CA were seen in less than 1.0% of healthy young adults (500) or patients without mononucleosis (500) submitted for heterophil studies. Unclassified CA were noted in 3.2% of the latter 1000 samples.


Induction of human granulocyte differentiation in vitro by ubiquitin and thymopoietin. Human bone marrow cells were separated according to density by centrifugation on Ficoll-Hypaque gradients and then according to size by velocity sedimentation. This procedure resulted in fractions enriched for immature granulocytes, mature granulocytes, and lymphocytes. Cells in these fractions were analyzed for their expression of certain surface and functional differentiation markers and for their ability to respond to thymopoietin and ubiquitin with the expression of additional differentiation markers. A higher percentage of band form and segmented granulocytes than of more immature granulocytes expressed complement receptors on their surfaces. Thymopoietin and ubiquitin induced a significant percentage of the cells in the immature granulocyte fraction to express this marker. These data suggested that the complement receptor may be viewed as a differentiation marker on human granulocytes, the expression of which can be induced in vitro by thymopoietin and ubiquitin. Furthermore, fractions containing predominantly band form granulocytes were induced by ubiquitin (but not thymopoietin) to develop the capacity to respond to chemotactic agents, and cell fractions containing predominantly myelocytes and metamyelocytes were induced by thymopoietin and ubiquitin to develop the capacity to phagocytose latex particles. These findings indicated that thymopoietin and ubiquitin, two agents known to induce a number of stages of human and mouse lymphocyte differentiation, are also capable of inducing some stages of human granulocyte differentiation in vitro.


Memory performance after arousal from different sleep stages. Learning material was presented to independent groups of subjects either after arousal from non-Rapid Eye Movement (non-REM) sleep, after arousal from REM sleep, or under conditions of no prior sleep. Measures of immediate and subsequent free recall were taken. Memory performance was found to be impaired where learning took place after non-REM arousal. This was manifest in the number of categories recalled, over both immediate and subsequent recall, and in the number of items recalled per category over subsequent recall. It was suggested that the memory performance decrement after non-REM arousal may be understood in terms of a retrieval deficit as well as a coding deficit. It is possible that the former is consequent upon a lower general level of arousal, whereas the latter is specific to memory.


Evolution of poliovirus since introduction of attenuated vaccine. Genetic marker tests were performed on 997 strains of poliovirus isolated from patients with neurological disease and from healthy people in England and Wales. Before the introduction of live attenuated vaccine most strains could multiply at raised temperatures. Now, however, many strains isolated from cases of poliomyelitis or from healthy persons with no known contact with vaccine cannot grow above 37 degrees C, and in this respect resemble the vaccine strains. The three serotypes are also much more evenly represented. Hence probably to a limited extent the vaccine-like strains have established themselves in the community.


Estradiol and testosterone binding in normal and mutant mouse cerebellum: biochemical and cellular specificity. Estradiol and testosterone binding macromolecules are demonstrated in normal mouse cerebellum. Identity of the estradiol binder as the 'receptor' is provided by its binding to DNA-cellulose, which does not occur for other estradiol binding proteins. The androgen binder is identified as 'receptor' by its specific deficiency (85% reduced) in the androgen-insensitive mutant mouse, testicular feminization (Tfm). The relative amounts of these two components are reversed in cerebellum compared to hypothalamus-preoptic area. Neurological mouse mutants, which lack specified neurons, are examined to test for the cells in which these hormone binding proteins are located. Experiments with Purkinje cell degeneration (pcd), weaver (wv) and staggerer (sg), suggest that the majority of these receptors are present in granule cells.


Prostaglandin E2, cyclic adenosine monophosphate and morphine analgesia. Intravenous administration of prostaglandin E2 (PGE2) to mice results in a significant distribution of PGE2 into the brain and an elevation of cyclic adenosine monophosphate (cAMP) in the midhindbrain and corpus striatum. Neither acute morphine administration nor morphine tolerance alters this response. Morphine alone also elevates cAMP levels. Although these elevations are blocked by the narcotic antagonist, naloxone, they only occur at supra-analgesic doses. Therefore morphine analgesia does not correlate with either elevation of cAMP levels of antagonism of PGE2-induced elevations of cAMP.


Purkinje cell activity during motor learning. Monkeys were trained to grasp a handle and move it in a horizontal arc to a central position by flexing or extending the wrist. A torque motor applied forces to the handle that switched at random intervals to alternately load flexor and extensor muscles. At each load switch, the handle was displaced transiently from the central position, and then moved back by the monkeys and held there steadily again. Recordings were made from cerebellar Purkinje cells (P-cells) whose simple spike (SS) activity was related to the task. The magnitude of one of the oppositely directed loads was then altered and the monkeys took about 12-100 trials with the novel load before performing as regularly as previously. During this period with one known and one novel load, some P-cells underwent increases in complex spike (CS) frequency at specific times after the load switch. This increased CS frequency would last for a similar number of trials as that taken by the monkey to adapt to the novel load before decreasing to near its previous level. Associated with the increased CS frequency ther were decreases in SS frequency that persisted after the CS frequency had decreased to near its previous level. These results are consistent with theoretical proposals that motor learning takes place in the cerebellum through changes in the strength of transmission of parallel fiber synapses on P-cells caused by the climbing fiber input. These results further suggest that climbing fiber firing causes a decrease in the strength of parallel fiber synapses.


Autoradiographic localization of opiate receptors in rat brain. II. The brain stem. The distribution of opiate receptors, determined by the autoradiographic localization of stereospecific [3H]diprenorphine binding sites, was studied in the brain stem. Areas showing very dense or dense localizations of receptors included the parabrachial nuclei, the superior colliculus, the ventral median raphe nucleus, components of the accessory optic system, portions of the habenulo-interpeduncular complex, the pretectal nuclei and the ventral lateral geniculate, the infundibulum and the medial thalamus. Moderate grain densities were found in broad areas of the upper medulla, midbrain and diencephalon. The significance of these findings are discussed in terms of pain mechanisms, association of some receptors with small diameter axons, pupillary reflexes, hormonal control, and effects of opiates on neurotransmitter systems.


Effect of clofibrate treatment on chronic hyperlipidemia induced by an ACTH-producing tumor. Plasma triglyceride and cholesterol levels were significantly increased in rats bearing an MtT-F4 tumor for 31-41 days. Addition of clofibrate at a dose of 30 mg kg-1 to the diet of rats bearing the tumor resulted in the complete elimination of the tumor effect on the plasma triglycerides, and to a great extent prevented the rise in the plasma cholesterol.


Cyclic AMP and the positive inotropic effect of norepinephrine and phenylephrine. Time-response studies of the effects of norepinephrine and phenylephrine revealed that both agonists caused an increase in cyclic AMP levels before increases in contractile force in either the electrically stimulated left atria or spontaneously beating right atria of the rat. Norepinephrine caused a nearly sixfold increase in cyclic AMP, whereas phenylephrine produced only a 50% increase in the nucleotide. Pretreatment with reserpine did not affect the norepinephrine cyclic AMP response; however, the phenylephrine cyclic AMP response was abolished. Reserpine pretreatment did not significantly affect the contractile responses of either amine. In the presence of propranolol, norepinephrine was found to have the ability to produce an increace in contractile force in which cyclic AMP was apparently not involved. The time course of the contractile response induced by adrenergic amines was found to be remarkably influenced by the chronotropic response in spontaneously beating preparations while the cyclic AMP response was not greatly affected. This difference in the contractile response may be due to the ability of the chronotropic response to influence the flux of calcium through the cell membrane.


Pyocin sensitivity of pseudomonas aeruginosa pretreated with antibiotics. Strains of Pseudomonas aeruginosa exposed to subinhibitory concentrations of either polymyxin B, gentamicin, or carbenicillin were compared with untreated organisms for changes in sensitivity to pyocin. Pretreatment with polymyxin B or gentamicin resulted in a decreased sensitivity to pyocin, while carbenicllin pretreatment did not alter pyocin sensitivity.


The structure of cellulose-producing bacteria, Acetobacter xylinum and Acetobacter acetigenus. The structure of the pellicles and cells of the cellulose-producing bacteria, Acetobacter xylinum and Acetobacter acetigenus, was studied by transmission electron microscopy of thin sections and freeze-etch replicas of glucose-stimulated cell suspensions, quiescent cell suspensions, and discrete pellicles. These bacteria have a relatively thin cell wall in section, with several irregular features superimposed on an otherwise simple, Gram-negative morphology. There are no flagella or pili. Unfixed, unextracted cells, viewed as whole mounts, show spherical or ellipsoidal bodies of undetermined composition which disappear after extraction with water or ethanol and propylene oxide. For both species, there are several kinds of cell surface irregularities, some of which are localized protrusions of the cell envelope. A variety of irregularities is seen frequently on cells in the first minutes of glucose incubation, on cells in a discrete pellicle, on quiescent cells, and on starved cells. Immediately after the addition of glucose to cellulose-free cells in suspension culture, fine fibrils appear on and (or) near the cell envelope. The fine fibrils are frequently as small as 3 nm in diameter in both freeze-etch and thin-section preparations and are frequently associated with freshly synthesized cellulose fibrils. Starved cells in suspensions free of (classical) microfibrils sometimes reveal stubs of an extracellular structure whose morphology resembles that of a nascent cellulose fibril.


Carcinoembryonic antigen in cancer of the female reproductive system. Serial plasma values correlated with disease state. The results of plasma carcinoembryonic antigen (CEA) determinations done over 600 patients with gynecologic malignancy will be presented. It would appear from this extensive survey that the likelihood of a patient having a positive value is increased with advancing stage and bulk of disease. The incidence of positive values in patients with clinical recurrence is quite impressive and presents a possible mode of follow-up for patients after standard treatment techniques have been administered for cervical cancer. Most interesting is the effect of radiation therapy or surgery on squamous cell cancer of the cervix and vulva in patients who have a positive value of the onset. Treatment of the disease by either modality appears to be associated with a precipitous drop in plasma value of CEA. Unfortunately, the presence or absence of CEA is not reliable and to date it is impossible to predict which patients with gynecologic malignancy will manifest a positive plasma value. Comments will be made concerning retro-de-differentiation and de-repression as a mechanism for the production of this antigen in patients with gynecologic cancer.


Membranous basal cell adenoma of parotid gland, dermal cylindromas, and trichoepitheliomas. Comparative histochemistry and ultrastructure. A basal cell adenoma of parotid, eccrine dermal cylindromas and trichoepitheliomas occurring in the same patient were examined by light and electron microscopy and histochemistry. The eccrine and parotid adenomas were similar both structurally and histochemically except for the presence of Langerhans cells in the cutaneous adenoma and well differentiated mucinous cells in the parotid tumor. The three different hamartomas found in this individual may represent the effect of a single pleiotropic gene acting on ontogenetically related stem cells.


Extramammary Paget's disease of the vulva. A clinicopathologic study of 13 cases. The clinicopathologic findings of 13 patients having extramammary Paget's disease of the vulva are discussed with emphasis on its histogenesis and biological behavior. For the purpose of study and assessment of prognosis, these cases were divided into two groups, those with an underlying invasive cutaneous adnexal adenocarcinoma, and those lacking an underlying invasive lesion. Four cases contained invasive cutaneous adnexal adenocarcinoma; in one of these the invasion was superficial. Three of the cases with an invasive lesion and three other cases showed in situ adenocarcinoma of sweat glands. Surgical treatment is mandatory for both groups of patients. The prognosis was excellent for the patients having Paget's disease without an underlying invasive carcinoma. From the literature, the prognosis of those with an underlying invasive carcinoma of the vulva appears to be less favorable. Multiple surgical excisions may be required to control the recurrences and metastases. A frequent association with other internal malignancy was observed. In four cases, second malignancies were found. Of special interest was the demonstration in one case of columns of neoplastic cells extending from involved sweat glands to the surface epithelium via the intradermal sweat duct. Our study leads us to support the concept that the Paget's cells, in a number of cases, are derived from an underlying carcinoma in situ of sweat gland origin.


The relationship between nephroblastoma and neurofibromatosis (Von Recklinghausen's disease). Three cases with co-existent Wilm's tumor and Von Recklinghausen's Disease from a series of 342 nephroblastomas are discussed as to the significance of this association. Biological arguments favor a positive link because of the multiplicity of associative lesions which parallel each other, the co-existence of which suggests a common genetic bond. An embryologic association is described whereby early mutational events, i.e. neural induction of nephrogenesis, may link these two entities in utero. Further interconnection is stated statistically, in that nephroblastoma patients in this series had a 29 fold higher incidence of multiple neurofibromatosis than predicted for in the general population. The co-existent diseases in the three cases observed are approximately 33-50% more than can be accounted for by chance in the United States population. Malignant transformation in neurofibromatosis is discussed because of the therapeutic implications presented by two of the three Wilms' tumor patients studied.


Mucinous (adenocystic) carcinoma of sweat glands with widespread metastasis. Case report with ultrastructural study. An unusual case of mucinous adenocarcinoma of the sweat glands originating from the right axilla is described. Despite complete control of the primary tumor after local excision, diffuse metastatic lesions continued to appear in the scalp, face, upper and lower extremities, sacral and pelvic bones, and left posterior iliac bone marrow space. The tumors were radioresistant. Multiple trials of various chemotherapeutic regimens, including alkylating agents, antifolates, antipyrimidines, vinca alkaloids, and antineoplastic antibiotics, were ineffective. The cutaneous neoplasms were grossly round, smooth, red, glistening, and cystic and were filled with a gelatinous mucinous material. Microscopic and ultrastructural findings are described.


Bone-marrow examination in the staging of small-cell anaplastic carcinoma of the lung with special reference to subtyping. An evaluation of 203 consecutive patients. Histologic examination of bone-marrow from the posterior iliac crest was routinely done as a pretreatment staging procedure in 203 consecutive patients with small-cell anaplastic carcinoma of the lung. Subtyping of the patients according to the WHO classification included 27.8% with "fusiform" cell type (WHO II,1), 28.3% with "polygonal" cell type (WHO II,2), 42.8% with "lymphocyte-like" cell type (WHO II,3), and 1.1% with mixed types (WHO II,4). Bone-marrow involvement was found in 17.2%. No significant difference was observed among the histological subtypes with regard to bone-marrow involvement. A comparison of bone-marrow biopsy and aspiration in patients investigated with both procedures showed that aspiration alone was diagnostic in nine of 24 (38%) positive patients as compared with two of 24 (8%) with biopsy alone, while in the remaining 13 patients (54%) both procedures were positive. Of the 35 patients with positive bone-marrow examination, 77% had no other evidence of distant metastatic disease if liver metastases identified by peritoneoscopy and liver biopsy are excluded as a staging procedure. With the exception of thrombocytopenia which was observed in six patients, with bone-marrow metastases, hematological findings were of little value in detecting bone-marrow involvement.


Lung cancer type in relation to cigarette dosage. In a retrospective study of 1,228 white males with histologically confirmed bronchogenic carcinoma, the proportion of squamous cell carcinoma increased with increasing age at diagnosis. Since the distribution of cell types was much the same in the 73% of cases aged 50-69, the relationship of cancer type to daily cigarette dosage was studied in this group. Squamous cell carcinoma increased from 48% of those men who smoked less than 20 cigarettes per day to 61% of those who smoked 40 or more cigarettes per day. Comparison with other studies showed conflicting results.


Congenital hepatic fibrosis, liver cell carcinoma and adult polycystic kidneys. In reviewing the literature, we found no liver cell carcinoma (LCC) or well-documented adult polycystic kidneys (APK) associated with congenital hepatic fibrosis (CHF). We report a 69-year-old man with CHF, LCC, APK, duplication cyst of distal portion of stomach, two calcified splenic artery aneurysms, myocardial fibrosis and muscular hypertrophy of esophagus. The LCC was grossly predunculated and microscopically showed prominent fibrosis and hyaline intracytoplasmic inclusions in the tumor cells.


Double primary cancers in 2 young sibs, leukemia in another, and dextrocardia in a fourth. Two brothers developed multiple primary neoplasms in childhood; one had glioblastoma and non-Hodgkin's lymphoma at age 11 years, and the other brain tumor and acute leukemia at six years. A third brother died with myelogenous leukemia at thre years, and a fourth with cyanotic congenital heart disease at 11 weeks. Each child also had at least one hamartomatous lesion of the skin. The clinical features suggested von Recklinghausen's neurofibromatosis or other inherited cancer syndrome, but laboratory studies identified no markers of susceptibility to familial neoplasia.


Hepatoblastoma in infant sister and brother. Two infants, a sister with motor retardation and brother with slight microcephaly and an undescended testis, died of hepatoblastoma. Only another documented familial occurrence of this tumor, affecting siblings of the same sex, can be found in the literature. The two patients described in this paper exhibited high platelet counts prior to liver resection. Although iron deficiency may have contributed to the thrombocytosis, the finding of many megakariocytes within the hepatoblastomas suggests an intra-tumoral production of platelets. An epidemiological investigation of the family under study failed to yield conclusive data. Hepatoblastoma is a rare tumor, but it may affect more than one sibling. Therefore, periodic clinical and laboratory evaluations of the siblings at risk appear to be justified.


Differences in mouse mammary tumor viruses. Relationship to early and late occurring mammary tumors. The murine model has been used extensively to study the various factors involved in the etiology of mammary carcinoma. Inbred mouse strains have been classically categorized into (i) high incidence stains with tumors occurring relatively early in the life of the animal, or (ii) low or moderate incidence strains with tumors occurring later on in life. We have radioactively labeled the RNA genome of the mouse mammary tumor virus (MMTV) from each of several mouse strains. We report here, using the technique of molecular hybridization, that the class of MMTVs responsible for the early occurring mammary tumors in high incidence strains can be clearly distinguished from the MMTVs associated with late occurring mammary tumors in low or moderate incidence strains; we also demonstrate that minor differences in MMTV genomes can also exist within these classes. Our findings show that MMTVs are transmitted via the germ line (as a germinal provirus) in some mouse strains, whereas in other strains, a non-germ line transmission is clearly demonstrated. Biochemical techniques can thus be used to track the mode of transmission of oncogenic viruses. The relationship of these findings to an understanding of the etiology of mammary carcinoma is discussed.


Biology of breast preneoplasia.

Biology of breast preneoplasia. The literature dealing with mammary cancer and the presence of the identifiable preneoplastic lesions in the mammary gland of several species has been reviewed. A discrete morphologically identifiable lesion with a high malignant potential is considered preneoplastic. Lobuloalveolar lesions have been identified in the human mammary gland which fit most of the criteria for preneoplasia. The lesions, hyperplastic atypical lobules, are multicentric, have a high statistical association with breast cancer and demonstrate a morphological progression through dysplasia to frank carcinoma. Other breast lesions have a high statistical association but no morphological progression can be seen and their neoplastic potential remains unknown. The significance of these correlations is enhanced by the occurrence of homologous breast lesions in outbred and inbred animal models. Statistical correlation with breast cancer and aberrant morphology has been accepted as presumptive evidence of preneoplasia in some animals. Definitive proof of the  neoplastic potential has been obtained in rodent models by transplantation of the suspected lesions and their subsequent development into cancers. Although a strong morphological and statistical analogy exists between the human and animal models, the causes and biological potential of the human lesions await experimental proof.


The concept of minimal breast cancer and the pathologist's role in the diagnosis of "early carcinoma". Minimal breast cancer has been variously defined as a lesion 1 cm or less, or even 5 mm or less. Some authorities consider intraductal cancer and lobular carcinoma in situ as minimal, but we believe that these should be excluded from consideration. Minimal cancer is not early cancer inasmuch as a 1 cm lesion represents 30 doubling times, and a certain percentage of these small cancers have already disseminated. The pathologist's responsibility in diagnosing minimal cancer has become more difficult. His problem is in three major areas: Is this proliferative cystic lesion cancer? Is this lobular or intraductal proliferative lesion in situ carcinoma? And if so, he must be concerned also with the treatment as well as the diagnosis. In any screening program, these minimal cancers, either invasive or in situ, should be evaluated by a group of experts in order that the program be entirely accurate.