Benign tumors of the liver. Pathologic examination of 45 cases. The histologic and clinical features of 45 benign tumors of the liver (excluding hemangiomas) were reviewed. They included 21 cases of focal nodular hyperplasia, 11 cases of hepatic cell adenoma, 12 cases of bile-duct adenoma and a single case of mesenchymal hamartoma. By adhering to published histologic criteria, the cases of focal nodular hyperplasia and hepatic cell adenoma could be readily separated. Although both focal nodular hyperplasia and hepatic cell adenoma occurred predominantly in women, focal nodular hyperplasia occurred at a greater mean age, was usually asymptomatic, and the lesions tended to be smaller (surgical cases) than hepatic cell adenoma. The latter usually were symptomatic. It is emphasized that focal nodular hyperplasia and hepatic cell adenoma are distinct lesions and are separable on histologic grounds. Although the names focal nodular hyperplasia and hepatic cell adenoma are retained, the authors cannot assert that one lesion is a neoplasm, the other a reactive condition, and prefer, instead, to leave this question unresolved.


Müllerian adenosarcoma of the uterus. Ultrastructure before and after radiation therapy. A case of müllerian adenosarcoma arising in the endometrium was studied by light and electron microscopy before and after radiation treatment. The tumor was composed of malignant stroma containing undifferentiated mesenchymal cells admixed with mature fibroblasts and other cell that contained crystalloids. The surface epithelium was lined by epithelial cells with and without cilia, similar to the normal surface epithelium cells of the uterus. High-dose therapeutic irradiation eliminated the undifferentiated mesenchymal cells from the tumor but did not alter the morphology of other epithelial and mesenchymal elements. The immaturity of mesenchymal cells and their radiosensitivity coupled with the ultrasonographic evidence of growth of the tumor point to the probable malignant nature of this neoplasm.


Cystosarcoma phyllodes. A clinicopathologic study of twenty-six hypercellular periductal stromal tumors of the breast. A series of 26 hypercellular periductal stromal tumors of the breast, commonly referred to as cystosarcoma phyllodes, is reported. Twelve neoplasms were classified histologically as benign and 14 (54%) as maligant after microscopic assessment of the stromal elements. None of the histologically benign cystosarcomas metastasized, although two recurred locally. Malignant transformation was not observed in the recurrent tumors. Local recurrences developed in four women with histologically malignant cystosarcomas and caused the death of one by direct extension into the lung. Pulmonary metastases of the sarcomatous element occurred in three. Two of these women died and one has survived 16 years after pulmonary lobectomy. Separation of cystosarcomas into benign and malignant categories on the basis of their histopathologic features is advocated. It is likely that overgrowth of the sarcomatous stroma is a prerequisite for metastasis.


Decreased angiotensin-converting enzyme in the adult respiratory distress syndrome. Using hippuryl-L-histidyl-L-leucine as substrate, serum angiotensin-converting enzyme was measured in 13 patients who had adult respiratory distress syndrome, eight patients with respiratory failure without adult respiratory distress syndrome, and two groups of controls: 24 healthy blood donors and 24 hospitalized patients with a variety of conditions but without respiratory failure or adult respiratory distress syndrome. Serum angiotensin-converting enzyme expressed in units/ml was 14.60 +/- 5.60 for adult respiratory distress syndrome compared with 28.92 +/- 6.60 for the blood donors, 20.76 +/- 5.87 for the patients with respiratory failure without adult respiratory distress syndrome and 20.20 +/- 5.94 in the hospitalized patients without respiratory failure or adult respiratory distress syndrome. These differences were significant, P less than .001 when adult respiratory distress syndrome was tested against the blood donors and P less than .01 against the other two groups. The significance of these findings is not clear, but the possibility is raised that the decrease of angiotensin-converting enzyme in adult respiratory distress syndrome results from a loss of pulmonary endothelial cells, which are known both to produce angiotensin-converting enzyme and to be damaged in adult respiratory distress syndrome.


Oral and intrauterine contraception: a 1978 risk assessment. This report reviews the current literature on the various risks associated with the use of oral contraceptives and intrauterine contraceptive devices. Reports on oral contraceptives from large prospective studies are not beginning to supplement the detailed reports resulting from earlier case-control studies. These studies suggest that in Western societies there is an increase in the incidence of a variety of circulatory diseases, with an increased risk of death. With intrauterine contraceptive devices it now has been fairly well documented that there is an increased risk of pelvic inflammatory disease. After reviewing and assessing the risks of both methods in some detail and comparing these risks to the benefits, the resultant conclusion is that the benefits continue to outweigh the risks for both methods, except for older women who choose oral contraceptives and also are heavy smokers.


Phagocytosis by cells of the pulmonary alveoli. Transformation of crystalline particles. The progressive transformation of illite crystals in alveolar macrophages has been studied with x-ray microanalysis and electron microdiffraction. Illite particles captured by macrophages undergo a series of progressive modifications which affect their ultrastructure, crystalline state, and chemical composition. Crystalline structure progressively disappears and certain elements are eliminated. At the end of the experiment, which lasted more than 6 months, the initial characteristics of the particles were greatly transformed. The constitutive elements of the illite particles were also found in structures other than macrophages: Type I pneumocytes, interstitium, and blood platelets.


Cell biology of leukocyte abnormalities--membrane and cytoskeletal function in normal and defective cells. A review. In this review I have attempted to explain the processes of chemotaxis, phagocytosis, oxidant generation, and lysosomal degranulation in normal and genetically abnormal human PMN. In my view these leukocyte functions are most importantly dependent on the integrity of three cellular components: the plasma membrane, the submembranous microfilaments, and the cytoplasmic microtubules. These components are often discussed in isolation, and the biochemical and pharmacological aspects of their function are analyzed separately here. However, PMN motile and bactericidal activities require the interdependent functioning of membranes, microtubules, and microfilaments. I have therefore tried to provide an integrated view of cytoskeleton-membrane organization and function in human PMN. I have particularly emphasized dynamic aspects of the cytoskeleton and membranes, eg, the induction of microtubule assembly and membrane enzyme activation by surface ligands and the reorganization of microfilaments in response to the same ligands. With this background established, I have selected for discussion a series of diseases in which abnormalities of chemotaxis, phagocytosis, lysosomal degranulation, and/or oxidant generation can be explained directly or indirectly by abnormalities in dynamic properties of PMN membranes, microtubules, or microfilaments. I emphasize that even preliminary insight into the basis of these disorders has sometimes been sufficient to suggest useful clinical approaches to the management of patients. In several of these neutrophil abnormalities, ie, neutrophil actin dysfunction, Chédiak-Higashi syndrome, and its "antithesis" described by Gallin and co-workers, the cellular dysfunctions were well documented but the molecular basis was completely obscure prior to cell biologic analysis. Snyderman and Pike 159 and Chusid and co-workers 160 emphasized the existence of a large number of other neutrophil bactericidal abnormalities resulting from as yet unexplained cellular defects. Further analyses of the functional interactions between membranes and cytoskeletal components in neutrophils may not only clarify the molecular bases of the disorders described here but also may provide insight into the origins and proper therapeutic approach to other granulocyte dysfunctions.


Ultrastructure of nitrosoheptamethyleneimine-induced lung tumors in European hamsters. The ultrastructure of lung tumors induced in European hamsters through chronic subcutaneous treatment with nitrosoheptamethyleneimine (NHMI) is described. The neoplasms demonstrated either adenomatous (adenomas and adenocarcinomas) or mixed (tumors with areas of adenoid and squamous differentiation) pattern. Adenomatous areas consisted of Clara cells and of cells with lamellated inclusion bodies in their cytoplasm. Cells in areas with squamous differentiation contained cytoplasmic filaments and filament bundles as well as dense-cored granules.


Reactive forms of oxygen and chemiluminescence in phagocytizing rabbit alveolar macrophages. Chemiluminescence (CL), superoxide anion (O2-) production, and particle uptake were measured to determine the role of antibacterial substances in the chemiluminescent response associated with phagocytosis in rabbit alveolar macrophages (AM). Exposure of AM to zymosan particles induced both CL and the production of extracellular O2-. CL is inhibited by superoxide dismutase, an enzyme which catalyzes the conversion of O2- to hydrogen peroxide (H2O2), by catalase, an enzyme which destroys H2O2, and by the hydroxyl radical (.OH) scavengers, benzoate and ethanol. Superoxide dismutase and catalase probably exert their effects in the extracellular fluid. CL can also be produced by the addition of NaO2 or H2O2 to zymosan in a noncellular system. The chemiluminescent response occurs before particle uptake is complete, which also indicates that CL occurs in the extracellular fluid. These results suggest that CL induced by zymosan in AM is due to the extracellular reaction between various reactive forms of oxygen and zymosan.


Binding of aldosterone to cytoplasmic and nuclear receptors of the urinary bladder epithelium of Bufo marinus. Binding of aldosterone to cytoplasmic and nuclear sites in urinary bladder epithelia of Bufo marinus (Dominican variant) is saturable and dependent upon steroid concentration. Scatchard analysis of specific cytoplasmic binding yielded a maximal binding capacity (N) of 14.5 X 10(-14) mol/mg protein and an apparent equilibrium dissociation constant (Kd) of 1.4 X 10(-8) M. Since Scatchard analysis of specific nuclear binding was complex, this binding was resolved by a computer-generated cirve-fitting technique which analyzed total aldosterone bound. Nuclear binding was resolved into three sites: a nonsaturable site that was linearly dependent upon aldosterone concentration, and two saturable sites (types I and II). Type I sites had relatively low capacity for aldosterone (N = 31 +/- 1 X 10(-14) mol/mg DNA) and high affinity (Kd = 2.5 +/- 0.5 X 10(-9 M); tffininty (Kd = 8.6 +/- 1.7 X 10(-7) M). Competition for [3H]aldosterone binding by dexamethasone, corticosterone, cortisol, progesterone, testosterone, and 17 beta-estradiol demonstrated that type I nuclear sites have higher affinity for aldosterone than for other steroids. The findings are consistent with the inference that the type I site is the mineralocorticoid receptor.


Aldosterone receptor occupancy and sodium transport in the urinary bladder of Bufo marinus. The concentration dependence of binding of [3H]aldosterone to cytoplasmic and nuclear receptors was evaluated in urinary bladder epithelial cells of Colombian toads. One class of specific sites (sensitive to displacement by excess aldosterone) was detected in the cytosol. However, two classes of specific nuclear [3H]aldosterone binding sites were evident. In the nucleus, high-affinity (Kd = 2.7 X 10(-9) M), low-capacity (N = 15 X 10(-14) mol/mg DNA) sites (type I) were completely saturated at approximately 4 X 10(-8) M aldosterone, a concentration which gave a maximal increase in short-circuit current (SCC). Occupancy of low-affinity (Kd = 4.6 X 10(-7) M), high-capacity sites (N = 150 X 10(-14) mol/mg DNA) (type II) occurred at higher steroid concentrations and did not correlate with further increase in SCC. Binding parameters of Colombian and Dominican variants of Bufo marinus were compared. In both variants, the SCC increase elicited by aldosterone correlated with accumulation of type I complexes in the nucleus, but the relationship was markedly nonlinear. Various alternatives were considered as the basis for a curvilinear dependence of the increment in Na+ transport on abundance of nuclear type I complexes.


Sex difference in LH response to LHRH and DBcAMP and effect of testosterone. Because luteinizing hormone-releasing hormone (LHRH) stimulates both pituitary cAMP production and LH release, cAMP has been implicated in the action of LHRH on LH release. The effects of LHRH and DBcAMP on LH release were tested in 4-h incubations with pituitary cultures prepared from male or female rats. LH contents in medium and cells were separately determined by radioimmunoassays. LH release in response to 10 nM LHRH was significantly greater in cultures prepared from female rats (female-RPC) than in cultures prepared from male rats (male-RPC), 1,070 and 418% of control, respectively. Addition of DBcAMP (3, 5, or 10 mM) significantly stimulated LH release by female-RPC (212, 206, or 286% of control, respectively) but did not affect LH release in male-RPC. Furthermore, DBcAMP significantly increased the cellular LH content in female- but not in male-RPC. Testosterone pretreatment of female-RPC significantly lowered the LHRH-induced LH release but did not affect the DBcAMP-induced LH release. These data indicate that testosterone may contribute to the sex difference in pituitary LH response to LHRH but not to DBcAMP.


Influence of estrogen on renal vitamin D hydroxylases and serum 1alpha,25-(OH)2D3 in chicks. The influence of estrogen on the metabolism of 25-hydroxyvitamin D3 was studied in 2- to 5-wk-old chicks. Single injections of at least 500 microgram diethylstibestrol (DES) increased the conversion of 25-hydroxyvitamin D3 to 1alpha,25-dihydroxyvitamin D3 (1alpha,25-(OH)2D3) and suppressed the production of 24,25-dihydroxyvitamin D3 in chick kidney homogenates. Acute (one 5-mg) injections of testosterone or progesterone did not enhance the 25-hydroxyvitamin D3-1alpha-hydroxylase, indicating specificity. However, chronic pretreatment with DES appeared to allow the potentiation of previously unstimulatory steroids such as progesterone and testosterone. In addition, the hormonal metabolite of vitamin D3, 1alpha,25-(OH)2D3, was measured in 4- to 6-wk chick plasma after steroid treatment. Greater than 1 mg DES per day for 5 days was necessary to enhance the circulating level of 1alpha,25-(OH)2D3; testosterone alone had no effect. This elevation was rapid, occurring within 12--24 h after injection. These data suggest that estrogen (as evidenced by DES treatment) is a modulator of vitamin D metabolism along with other known regulators such as parathyroid hormone, phosphate, and 1alpha,25-(OH)2D3. The mechanism of the regulation is as yet unknown.


Cardiovascular responses to blockade of angiotensin and alpha-adrenergic receptors. Both angiotensin and alpha-adrenergic blocking agents reduce arterial blood pressure in hypovolemic states. We have compared the effects of an angiotensin antagonist (saralasin) and an alpha-adrenergic blocking agent (phenoxybenzamine) in supramaximal dosage on cardiac output, total peripheral resistance, and venous tone in rabbits rendered hypovolemic by restriction of sodium intake, supplemented by a furosemide-induced diuresis 48 h prior to study. Saralasin (10 microgram/kg per min) reduced arterial blood pressure significantly (-15 +/- 1.2 mmHg) despite an unchanged cardiac output (P less than 0.025) due to a fall in total peripheral resistance. Phenoxybenzamine (5 mg/kg) induced a much larger fall in arterial blood pressure (-28 +/- 3.6 mmHg), despite an identical reduction in total peripheral resistance, because cardiac output also fell (+/- 9 ml/kg per min). The reduction in cardiac output was associated with a significant increase in hindlimb venous distensibility (P less than 0.001) after alpha-adrenergic blockade. Saralasin, conversely, had no influence on venous tone. Adrenergic mechanisms contribute to cardiovascular homeostasis through an influence on both arteriolar and venous tone, whereas the effect of angiotensin is directed entirely to the arteriolar side of the circulation.


Distribution along the rat nephron of three enzymes of gluconeogenesis in acidosis and starvation. Methods were devised or modified which made it possible to measure phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase, and glucose-6-phosphatase in seven defined parts of single nephrons and in patches from thin limb and papilla areas dissected from freeze-dried microtome sections of rat kidney. All three enzymes were essentially confined to the proximal tubule. In normal kidneys, the levels were highest in the proximal convoluted tubule. Glucose-6-phosphatase was 20 times higher in the early part of the convoluted segment than in the late part of the straight segment. With one exception, in acidosis, only phosphoenolpyruvate carboxykinase increased (fourfold in the proximal convoluted segment but much less in the straight portion). In starvation, phosphoenolpyruvate carboxykinase increased about as much as in acidosis in the proximal straight tubule, but not as much in convoluted portions, whereas glucose-6-phosphatase rose modestly in both parts of the proximal tubule and fructose bisphosphatase rose only in the straight tubule, especially the early segment. It is suggested that ammoniagenesis can accompany gluconeogenesis in the proximal convoluted tubule but not in the straight segment.


Cholinergic-adrenergic interactions on intestinal ion transport. The autonomic control of intestinal electrolyte transport has been investigated in the in vitro, short-circuited rabbit ileum with varying doses of carbachol and with neuroeffector blocking agents. Low-dose carbachol (less than 10(-6) M) and high-dose carbachol (greater than 10(-4) M) had different effects on Na and Cl transport. Low-dose carbachol caused a transient increase in the potential difference and short-circult current, stimulated Cl secretion, and inhibited the residual flux (probably HCO3 secretion). This is a muscarinic response since it is inhibited by atropine (10(-6) M). After an initial increase of the potential difference and short-circuit current, high-dose carbachol reduced these electrical parameters, stimulated Na and Cl absorption, and abolished the residual flux. This is a nicotinic response since it is inhibited by hexamethonium (10(-5) M). This nicotinic response is identical to that reported by others with alpha-adrenergic agents and it was inhibited also by phentolamine (10(-7) M). We propose that high-dose carbachol stimulates nicotinic receptors on postganglionic sympathetic fibers present in our preparations causing a release of catecholamines and a resulting alpha-adrenergic response by the intestinal epithelial cell. The physiological significance of this response in the gut remains to be determined.


Calcium transport across segments of the rabbit distal nephron in vitro. The mechanism of Ca2+ transport by various segments of the distal nephron was studied in vitro using the isolated perfused tubule technique. Calcium absorption in the distal convoluted tubule (DCT) and the granular portion of the cortical collecting duct (CCTg) was significantly enhanced in the presence of parathyroid hormone (PTH), 3 X 10(-2) U/ml. Na+ was absorbed from and K+ was secreted into the lumen of the DCT. The presence of amiloride (5 X 10(-5) M) or furosemide (5 X 10(-5) M) in the perfusate of DCT each caused a partial inhibition of Na+ but not Ca2+ absorption. The foregoing result with Na+ is consistent with the heterogeneous nature of DCT. Net Na+ absorption and K+ secretion also occurred in the CCTg; both processes were completely inhibited by amiloride. Ca2+ absorption occurred in the thick ascending limb of Henle's loop; it was not enhanced by PTH, and the results were consistent with passive movement. No net Ca2+ movement was observed in the nongranular (light) segment of the cortical collecting tubule in the presence or absence of PTH or dibutyryl cyclic adenosine monophosphate.


[A simple modification for the electrochemical oxygen analyzer to improve accuracy during prolonged closed system anaesthesia (author's transl)]. A simple modification of the condenser used with the electro-chemical oxygen analyzer (Dräger BioMarine OA202 R) consists of filling the distal chamber with hygroscopic silicagel. This prevents condensation of water vapour on the sensor thus allowing the accuracy during prolonged closed system anaesthesia to remain nearly unaffected (fade less than 0.5%/h).


Antagonism of polymyxin B-induced neuromuscular and cardiovascular depression by 4-aminopyridine in the anesthetized cat. In an attempt to find a better antidote to polymyxin B-induced neuromuscular blockade, the authors tested 4-aminopyridine, 0.4--1 mg/kg, in ten anesthetized cats. The sciatic--tibialis anterior nerve--muscle preparation was used. The neuromuscular blockade was successfully reversed in all cats. Rapidity of reversal depended on the dose of 4-aminopyridine administered. At 0.6--1 mg/kg, reversal of the twitch response from 20 per cent of control to 80 per cent of control required 2.4 (SE, 0.5) min; to 100 per cent, 14.4 (SE, 3.7) min. An overshoot of the recovery of neuromuscular blockade of 6--10 hours' duration followed the reversal. The mechanical twitch response reached a peak of approximately 140 per cent of control at 2 hours. At a lower dosage, 0.4 mg/kg, two of three cats took more than an hour for complete recovery of the twitch response, and the overshoot was approximately 10 per cent of control. The hypotensive effect of polymyxin B was partially reversed, but the bradycardia was not. It is concluded that 4-aminopyridine is an effective antidote to polymyxin B-induced neuromuscular blockade in the cat.


[Association of angiosarcoma of the liver and hepatoma in vinyl chloride worker]. This report deals with a 51 years-old man who, for 23 years and 4 months of his working life, had been exposed to vinyl chloride vapors. Autopsy revealed a hepatoma associated with an angiosarcoma of the liver. The case is the first ever to be reported in the medical literature. This case raises doubts about the theory which suggests that the carcinogenic effect of vinyl chloride in man elicit a tumor of vascular nature.


Use of the double-isotope, single-injection method for estimating renal function in normal and polybrominated biphenyl-exposed dairy cows. A method is described for conducting a rapid and efficient renal function test in dairy cattle. The method, adapted from methods used for man and dogs, utilizes radiolabeled 131I-sodium iodohippurate to determine effective renal plasma flow (ERPF) and 125I-sodium iothalamate to determine glomerular filtration rate (GFR). The mean GFR for adult cattle was determined to be 2.8 +/- 0.4 ml/kg/minute with a biological half-life of 35.5 +/- 1.4 minutes. The mean ERPF was found to be 10.5 +/- 1.9 ml/kg/minute with a half-life of 17.9 +/- 0.6 minutes. These values are comparable with those in man, but are lower than values in dogs. A toxcity study was done with dairy cattle exposed to polybrominated biphenyls (PBB). Efforts were made to determine the amount of time required for kidney lesions to develop and, if possible, to delineate the potential site of action of PBB. Apparently PBB do not affect GFR or ERPF, even though they produce nephrotoxic effects. Potential mechanisms to explain these results are described.


Effect of experimental infection with pseudorabies (Aujeszky's disease) virus on pigs with maternal immunity from vaccinated sows. Live-virus and inactivated-virus vaccines were used to immunize sows against pseudorabies (Aujeszky's disease) virus. To test the efficacy of the vaccination, 53 pigs of different ages were taken from the 1st and the 2nd litters of vaccinated sows and placed separately in isolation units. The pigs were challenge exposed with virulent pseudorabies virus and examined for clinical signs, virus excretion, and serologic reaction. The challenge inoculum caused severe nervous or respiratory signs of disease in 12 of the 13 control pigs, with a mortality of 76%. The pigs from the 1st litters of sows vaccinated with the live-virus vaccine did not become sick, whereas 2 of the 9 pigs (22%) from the 2nd litters had clinical signs and died of pseudorabies. All pigs from sows vaccinated with the inactivated-virus vaccine remained healthy. The results of virus isolation from oronasal swabs, combined with the serotest results, indicated that challenge exposure of all except 1 of the pigs resulted in a subclinical infection with the formation of active immunity.


Virus isolation and immune responses in susceptible swine exposed with pseudorabies virus (Shope strain). Eighteen seronegative swine weighing from 9 to 11 kg were exposed intranasally with the Shope strain of pseudorabies virus (PRV) and were observed for 21 days in an experiment to detect virus shedding and immune responses. All swine had PRV in their nasal passages at 7 days after exposure; they also had precipitating antibodies to PRV as determined by the microimmunodiffusion test (MIDT) and very low levels of virus-neutralizing (VN) antibodies. The PRV was isolated from only 2 swine at postexposure day 14; all swine were MIDT positive, and VN titers ranged from 4 to 128. Virus was not isolated from the swine at 21 days after exposure, but all were MIDT positive; VN titers ranged between 8 and greater than or equal to 256.


Experimental infectious bovine rhinotracheitis virus infections of English ferrets (Mustela putorius furo L). Intranasal and intraperitoneal exposure of English ferrets (Mustela putorius furo L) to infectious bovine rhinotracheitis virus caused acute and chronic infections of the respiratory tract. The clinical syndrome was characterized by sneezing, coughing, and anorexia from postexposure days (PED) 3 to 7. Mucopurulent exudate was observed in the posterior nares and pharyngeal area of ferrets euthanatized on PED 4 and 8. The virus was readily recovered from the turbinates, respiratory tract epithelium of the pharynx, retropharyngeal lymph nodes, trachea, lungs, and spleen of animals euthanatized on PED 4, but only from the respiratory tract epithelium of the pharynx in ferrets euthanatized on PED 8 and 12. Results of histopathologic studies revealed an acute suppurative pharyngitis in animals euthanatized on PED 4 and 8. Recrudescence of chronic infection could be elicited by daily intraperitoneal injections of 4.0 mg of dexamethasone. However, daily administration of 2.0 mg of dexamethasone intraperitoneally did not cause more severe clinical disease. Results of serologic studies revealed serum antibody profiles comparable with those expected in experimentally exposed cattle.


Neural changes in recurrent infection of infectious bovine rhinotracheitis virus in calves treated with dexamethasone. Recurrent infection by infectious bovine rhinotracheitis (IBR) virus was induced in calves by dexamethasone (DM) treatment (given 5 days) at 5 months after primary infection. The virus appeared in nasal secretions of the calves on the 4th day after initiation of DM treatment and continued until the 9th day. The calves were killed on the 1st, 3rd, 4th, 5th, 6th, 7th, 8th, 10th, and 11th days after DM treatment was started for examination by histopathologic and immunofluorescent antibody techniques. The most significant neural change was trigeminal ganglionitis with neuronophagia, which was observed from the 3rd to the 11th day. Significantly, the extent of changes in the trigeminal ganglion and medulla oblongata corresponded to the amount of DM treatment administered. The IBR virus antigen was first observed in the trigeminal ganglion cells, and thereafter, it was detected in the Schwann cells, satellite cells, neuroglia cells, and nasal mucosa until the 10th day. These observations indicate that the IBR virus is capalbe of producing a persistent infection in the trigeminal ganglion and that trigeminal ganglionitis may be a characteristic lesion for inducing the reactivation of lagent IBR virus.


Immune responses in swine given lipid-conjugated pseudorabies viral antigens. The immune response was compared in pigs given inactivated pseudorabies virus (PRV) antigens (with or without adjuvant) or PRV antigens covalently conjugated with a fatty acid (lauric acid) to enhance delayed-type hypersensitivity. The pigs were given 2 inoculations, 14 days apart, and were challenge exposed 28 days after the 1st inoculation. Pibs inoculated with PRV antigens, with or without adjuvant, had significant virus-neutralizing (VN) antibodies before challenge exposure, but the pigs inoculated with lipid-conjugated PRV antigens had no detectable VN antibodies, with the exception of 1 pig. All inoculated pigs were positive by the microimmunodiffusion test at postinoculation day 14 and remained positive throughout the experiment. The inoculated pigs had delayed-type hypersensitivity reactions when skin tested a postinoculation day 25; the pigs inoculated with lipid-conjugated PRV antigens had a more pronounced reaction. Inoculated pigs had mild respiratory signs on the 3rd through the 6th days after challange exposure, with no observable difference in severity between the inoculated groups. The control pigs had acute signs of PRV, and 3 or 4 pigs died 5 to 8 days after challenge exposure. The average VN titers of the different inoculated groups of pigs were nearly equal 2 weeks after challenge exposure. Results indicated that both humoral antibodies and cell-mediated immunity have a role in PRV infections in swine.


Colostral IgA, IgG, and IgM-IgA fractions as fluorescent antibody for the detection of the coronavirus of transmissible gastroenteritis. Colostrum from sows and gilts inoculated with virulent transmissible gastroenteritis virus was fractionated into the 3 major immunoglobulin classes, IgA, IgG, and IgM-IgA fractions, by chromatographic and gel-filtration procedures. Each fraction was assayed for purity with rabbit anti-porcine serum and rabbit monospecific anti-porcine IgG, anti-porcine IgA, and anti-porcine IgM. These analyses showed that the IgG and IgA fractions were pure. The IgM fraction contained some IgA in the polymeric form and was designated the IgM-IgA fraction. Each Ig was assayed for virus-neutralizing activity on swine testes cells by the plaque-reduction method before and after conjugation with fluorescein isothiocyanate. On the basis of activity per milligram of protein, the virus-neutralizing titers were 1:641, 1:44, and 1:6.8 for the IgA, IgG, and IgM-IgA fractions respectively; the fluorescent antibody titers were 1:31.3, 1:0.1, and 1:15.6, respectively, for the same Ig.


Secretory antibodies against turkey coronaviral enteritis. Studies on local immunity to transmissible coronaviral enteritis of turkeys (bluecomb) was made. Intestinal secretions and bile from affected birds contained secretory immunoglobulins against coronaviral antigen throughout the 6 months' duration of the experiment. Attempts to purify and to characterize the globulins in intestinal secretions and bile of the affected birds were made, using the techniques of gel filtration, DEAE chromatography, and immunoelectrophoresis. Class-specific anti-turkey IgA antiserum in the agar gel precipitin test further established the presence of IgA in the intestinal secretions and bile.


Mortality after long exposure to cummingtonite-grunerite. Ore containing cummingtonite-grunerite has been mined to extract gold since 1876 in Lead, South Dakota. Each of the 1,321 men who were recorded as having worked 21 years or more with the Homestake Mine was allocated to one of 5 dust-exposure categories on the basis of work history and available information on environmental conditions. All except 9 men were traced to the end of 1973, when 652 were still living; the cause of death was ascertained for 657 of the 660 men who had died. Deaths from cerebrovascular accidents and malignant disease were close to the numbers expected and from accidents and other causes were fewer than expected, but in each of the 3 diagnostic groups--pneumoconiosis (mainly silicosis), tuberculosis, and heart disease--there were more than 30 excess deaths. A clear dust-exposure relationship was found for pneumoconiosis and respiratory tuberculosis--with relative risks for the 2 groups with greatest exposure to dust as compared to the 2 with least exposure, of 19.9 and 16.0, respectively, but there was no convincing evidence of an increase in respiratory cancer.


[Kinetics of production of collagenase, esterase and elastase by human and soil strains of "Entomophthora coronata" (author's transl)]. A comparative study was made of the production of extracellular proteolytic enzymes, which were originated from two different strains, one pathogenic and one saprophytic, of E. coronata in stable culture at 30 degrees C. Quantitative differences were found: the maximal activity of the three enzymes (collagenase, esterase, elastase) produced by the strain isolated from soil was always lower than that observed with the human strain.


[Assay of human anti-exoproteins to group A streptococci by microhaemagglutination: correlation with anti-streptolysin O and various anti-enzymes (author's transl)]. A microhaemagglutination test in disposable U plates has been devised for rapid, quantitative evaluation in antistreptococcal antibodies in human sera. Fresh or freeze-dried glutaraldehyde-treated sheep erythrocytes sensitized with over fifteen extracellular proteins released by group A Streptococcus pyogenes including streptolysin O, deoxyribonucleases, hyaluronatelyase, streptokinase and nicotinamide dinucleotide glycohydrolase were used. Haemagglutination and anti-streptolysin O (ASLO) titers were determined in parallel on 434 serum specimens from 123 healthy subjects ("controls") and 311 patients with a history of supposed or evident streptococcal infection. The titration of the four above-mentioned anti-enzyme antibodies has also been made on about 100 sera from both groups. Haemagglutination titre (HT) was less than 800 in control sera. By contrast it was greater than 800 up to 12 800 in patients specimens. Very good correlation was found between HT on the one hand and ASLO or anti-SK, anti-HA and anti-NADase antibodies on the other hand. HT and anti-DNase B antibodies were less correlated. Haemagglutination titres appear to rise earlier than serological titres of conventional streptococcal antibodies. The haemagglutination test described may be particularly helpful as a rapid serologic indicator of streptococcal infections and more reliable than the titration of ASLO alone, or of any one of anti-enzyme antibodies.


The cells of origin of ACTH in man.

The cells of origin of ACTH in man. The evidence from ACTH-producing tumors, from the morphologic effects of excess glucorticoids on the hypophysis, and from immunocytochemistry all points to the predominant "basophils" of the human anterior lobe, the intensely PAS-possitive beta(R) cells, as the source of ACTH. Similar cells often also occur, sometimes in large numbers, in the pars nervosa. beta(R) cells can be immunostained with antibodies against alphaMSH and betaMSH, but it is likely that they actually contain the precursor molecule(s) of betaMSH, betaLPH (and may be gammaLPH). Once this is satisfactorily documented, the functional term corticolipotrops should replace the provisional name beta(R) cells. Electron microscopically, these cells contain randomly arranged granules with a maximum diameter of 300-500 nm, and occasionally small amounts of filamentous material, which increases dramatically in response to excess glucocorticoids. In the beta (R) cells of adults, a single set of granules stains for ACTH and with antibodies to betaMSH. In fetal life, ACTH cells appear early and material reactive with anti-betaMSH accumulates in them (and possibly also in cells not containing ACTH) only later. The posterior lobe beta(R) cells can be immunostained with both anti-betaMSH and antibodies against the COOH-terminal portion of ACTH, but the presence of bioactive ACTH in them remains to be shown.


[Rapid virus diagnosis by electrosyneresis (author's transl)]. Electrosyneresis (counter-current immunoelectrophoresis) detected the presence of 7 coxsackieviruses and 1 poliovirus in 70 specimens of stools. Identical results were obtained by inoculation of mice and tissue cultures. First screening of specimens was undertaken with a human "standard" gammaglobulin; only specimens precipitating with this reagent contained viral antigens. Hence, especially in children, all negative stools could be excluded from subsequent tests with antiserum pools and monospecific antisera. In our experience, electrosyneresis is a specific method: every type of coxsackievirus and poliovirus types react only with the corresponding monospecific antisera. Electrosyneresis appears to be a sensitive procedure: viral antigens corresponding to 10(4) TCID50/ml can be dtected.


[Unmasking effect of trypsin on Sendai virions obtained in KB cell cultures (author's transl)]. The infectivity of virus grown in KB cell cultures can be directly activated by trypsin. Thus the fit conditions for seeding are effected from subculture to subculture if the cell inoculum is pretreated by trypsin.


Assays and dynamics of corticotropin-releasing factor activity in rat hypothalamus. Features of several assays for corticotropin-releasing factor (CRF) were compared, with reference to sensitivity, precision, specificity and convenience. In general, in vivo assays are less specific, whereas in vitro assays with isolated pituitary cells appear to suffer from vulnerability. Promising approaches for overcoming this difficulty are appearing: one is the cell culture technique and the other is the perfusion of pituitary cell column. However, the findings with vasopressin are again discordant. So far, the in vivo-in vitro assay system appears to be the most satisfactory. In this connection, special emphasis was laid on the CRF assay by out intrapituitary injection technique through the parapharyngeal approach, which has features of both in vivo and in vitro systems. The drawbacks with this assay are its complexity and relatively lower precision. Problems of the use of dexamethasone are discussed from a viewpoint of possible multistage feedback inhibition, as postulated by Yates et al. Several findings on the CRF dynamics were mentioned, in order to exemplify the findings obtained with our assay: it was shown that the two-peaked changes of CRF after ether-laparotomy stress in adult rats were composed of heterogeneous components, as revealed by differential effect of cycloheximide. Pretreatment with cycloheximide similarly abolished the delayed CRF peak that was observed in 2-day-old neonatal rats under stress. These findings suggest an operation of a biochemical denominator in common with both cases.


Neural control of ACTH release in response to hemorrhage. A three dimensional reconstruction of the central neural pathways that appear to mediate release of ACTH in response to hemodynamic change is illustrated in Figure 11. Fibers from receptors in the right atrium and the carotid arteries project to the lateral solitary nucleus and then to the medial and the lateral nucleus intercalatus. A pathway containing projections from these nuclei then converges dominantly in the locus subcoeruleus and locus coeruleus. Multiple pathways then diverge, to travel in part directly to the hypothalamus through dorsal pathways. One pathway inhibits and another facilitates the release of ACTH. Multiple pathways also diverge, to travel in part medially, and then to the hypothalamus through ventral pathways. Again, one pathway inhibits and another facilitates the release of ACTH. The dorsal and ventral inhibitory pathways appear to converge in a region extending from just caudal and ventral to the paraventicular nucleus to the posterior hypothalamic area. Thus, after the coalescences of the various pontine-hypothalamic pathways, three principal pathways remain. These include a posterior inhibitor path, an anterodorsal facilitatory path that terminates in the paraventricular nucleus and that may be mediated through release of vasopressin, and an anteroventral facilitatory path that terminates in the suprachiasmatic and ventromedial nuclei and that is probably mediated through release of corticotropin-releasing hormone. The mode of integration of these pathways has not been defined. The pathways described herein are oligosynaptic: a signal may travel from atrium to hypothalamus over three to seven neurons. The combination of control of input hemodynamic signals and of measurement of ACTH permits quantitation of both sensory and motor events, that inevitably must be embedded in the neuronal pathways described here. The analysis of the input-output relations and their correlation with internal neural events must form the basis of a description of the physiology of the physiology of the system whose central neural anatomy has been defined in part by these studies.


Plasma ACTH levels in stressed and nonstressed adrenalectomized rats. Following adrenalectomy there is a triphasic response of ACTH secretion characterized by an initial high rise immediately following the adrenalectomy, secondary to the stress of the procedure. This is followed by a fall to resting ACTH secretion until a steady state is reached 1 to 2 weeks following adrenalectomy at levels about 10 times normal. The ability to respond to stress with increased ACTH secretion is not lost at any time following adrenalectomy.


The nutritional epidemiology of cardiovascular disease. A basic review of the extensive literature focusing on the major risk factors of atherosclerotic coronary heart disease and stroke, i.e., elevation of blood lipids related to diet, blood pressure elevation, and genetic factors using the traditional epidemiological model of interaction between host, agent, and environment, has strongly supported the concept that diet and particularly saturated fat and/or cholesterol are significant contributors to the elevation of blood lipids, especially cholesterol, and contribute importantly to the premature development and mortality of atherosclerotic coronary heart disease. Certainly genetics exert an important impact on this process. To date it remains unclear whether or not major changes in the dietary pattern of huge population groups can be practically effected. The minor dietary modifications so far studied in the average atherosclerosis-prone population cannot be anticipated to make a major dent in the epidemic proportions of atherosclerotic coronary heart disease. It is quite clear that prospective preventive medicine must be implemented at a very early age in the pediatric age group, in which atherosclerosis is now recognized by many as the number one pediatric problem. Tremendous biochemical advances have provided new insights in knowledge regarding the transport of blood lipids, particularly cholesterol, and the regulatory mechanisms at the cellular level for cholesterol under normal circumstances and in the genetic influenced hyperlipidemias (TABLE 4). A bright future lies ahead for the reduction of the epidemic of atherosclerosis which could be greatly enhanced by a greater personal responsibility for health care and a much more careful and prudent diet selection and exercise managment.


Adult cytomegalic inclusion retinitis.

Adult cytomegalic inclusion retinitis. Two cases of adult cytomegalic inclusion retinitis are described in renal transplant patients. The areas of initial involvement by the retinitis were left atrophic and demarcated from normal retina by a hypopigmented area in the retina. Both cases were complicated by the development of a secondary glaucoma with open angles. The glaucoma in each patient was successfully managed by carbonic anhydrase inhibitors, epinephrine drops, and cycloplegias. One patient still retains normal visual acuity in the eye which had active retinitis.


Ultrastructure and biochemistry of sympathetic ganglia in idiopathic orthostatic hypotension. The role of transsynaptic dysfunction in the pathogenesis of idiopathic orthostatic hypotension (IOH, or idiopathic autonomic insufficiency) was examined microscopically and biochemically in autopsy specimens. Light microscopy of the sympathetic ganglia showed abnormalities in all 4 IOH patients, including focal phagocytosis of neurons, increased numbers of satellite cells, and perivascular lymphocytic infiltrates. Electron microscopy revealed proliferation and hypertrophy of satellite cells and abnormalities in the unmyelinated axons. In contrast, the spinal cord intermediolateral columns, containing the presynaptic neurons, were unremarkable in 1 patient, exhibited only mild gliosis in another, and showed neuron loss and fibrillary gliosis in 2 patients. Postsynaptic dopamine-beta-hydroxylase (DBH) activity was decreased at least fourfold (p less than 0.02) in sympathetic ganglia of patients with IOH, while tyrosine hydroxylase (T-OH) was normal. Ganglion choline acetyltransferase (ChAc) activity, an index of presynaptic function and integrity, was normal in the IOH group. A number of our observations suggest that presynaptic disease is not an absolute requirement for adrenergic abnormalities in IOH. The intermediolateral columns of the spinal cord were histologically normal in 2 of the patients with IOH, and ultrastructural abnormalities in sympathetic ganglia were consistent with primary adrenergic degeneration. In addition, presynaptic ChAc activity was normal in IOH ganglia, whereas postsynaptic DBH activity was depressed. Finally, postsynaptic T-OH activity, which is regulated by transsynaptic mechanisms, was normal in IOH ganglia.


Increased sodium plus potassium adenosine triphosphatase activity in erythrocyte membranes in Huntington's disease. Dopa-decarboxylase, acetylcholinesterase, sodium plus potassium stimulated adenosine triphosphatase (Na+ + K+-ATPase), and membrane-bound protein kinase were compared in the erythrocytes of patients with Huntington's disease and normal controls. All these enzymes also exist in the basal ganglia. The Na+ +K+-ATPase level was elevated (p less than 0.05) in Huntington's disease, while no significant changes were observed in the other enzymes. This finding is consistent with the concept that Huntington's disease is associated with a general membrane abnormality.


Acetylcholine release in diaphragm of rats with chronic experimental autoimmune myasthenia gravis. Recent evidence indicates that in chronic experimental autoimmune myasthenia gravis (EAMG) and in human myasthenia gravis, the defect of neuromuscular transmission results from immune-mediated destruction of post-synaptic membrane at the neuromuscular junction, with a reduction in the density of acetylcholine (ACh) receptors and decreased sensitivity to ACh released by nerve impulses. In the present study, the amount of ACh released by nerve impulse in rats with chronic EAMG and control rats of the same age, weight, and sex was compared. Phrenic nerve-hemidiaphragm preparations were stimulated in vitro, and the amount of ACh released was measured by bioassay. Despite a marked reduction in the amplitude of miniature end-plate potentials in chronic EAMG, ACh output at rest and during stimulation was not different from that of control rats. These data support the concept that the defect of neuromuscular transmission is due to a reduction of postsynaptic sensitivity to ACh.


Pharmacological studies of the pupils in familial amyloid polyneuropathy. Pharmacological studies of the pupils of 7 patients with familial amyloid polyneuropathy (FAP) were performed in order to demonstrate the site of the lesion. Supersensitivity to 1.25% epinephrine was observed frequently, but not to 2.5% methacholine. The lesions of the sympathetic nervous system, judging from the epinephrine and tyramine tests, were regarded as preganglionic in 2 patients and postganglionic in 4. Among 3 patients reexamined six months later, 1 had developed signs of sympathetic postganglionic disturbance and another showed a peripheral parasympathetic disturbance associated with progress of a sympathetic preganglionic disturbance. From these results it was suggested that in FAP, sympathetic abnormalities involve the pupils more often than parasympathetic disturbances, and pharmacological studies can detect the autonomic dysfunction before it becomes clinically apparent.


Angiotensin converting enzyme in kainic acid--injected striata. The activity of angiotensin converting enzyme is decreased in kainic acid-injected neostriata of rats, providing another biochemical similarity between this animal model and Huntington's chorea.


System for evaluating clostridial inhibition in cured meat products. A method for evaluating inhibition of Clostridium botulinum, C. sporogenes, and C. perfringens in cured meat products was developed. This system can easily be used in the microbiology laboratory using aluminum ointment tubes as the product container. Swells caused by gas production by the organism are easily observed by using the aluminum tubes. Results obtained confirmed earlier work on the inhibitory effect of sodium nitrite and sorbic acid against the clostridia in cured meat products.


Survey of human virus occurrence in wastewater-recharged groundwater on Long Island. Treated wastewater effluents and groundwater observation wells from three sewage recharge installations located on Long Island were assayed on a monthly basis for indigenous human enteroviruses and coliform bacteria for a period of 1 year. Viruses were detected in groundwater at sites where recharge basins were located less than 35 feet (ca. 10.6 m) above the aquifer. Results from one of the sites indicated the horizontal transfer of viable viruses through the groundwater aquifer.


Poliovirus removal from primary and secondary sewage effluent by soil filtration. Adsorption of poliovirus from primary sewage effluent was similar to that from secondary sewage effluent in both batch soil studies and experiments with soil columns 240 cm long. Virus desorption by distilled water was also similar in a soil column that had been flooded with either primary or secondary effluent seeded with virus. These results indicated that absorption of poliovirus from primary effluent and virus movement through the soil were not affected by the higher organic content of primary sewage effluent.


Association of Yersinia enterocolitica with the manufacture of cheese and occurrence in pasteurized milk. Raw milk in southern Ontario frequently contains Yersinia enterocolitica. The potential for transmission of this organism by cheese manufactured from unpasteurized milk was evaluated by examination of milk and cheese curd samples from cheese manufacturing plants and finished cheddar and Italian cheeses. The incidence of Y. enterocolitica was lower in cheese curd samples (9.2%) than in raw milk (18.2%). Most of the curd samples showed a positive phosphatase test, indicating production from raw milk. One curd sample yielded Y. enterocolitica after 4 weeks of storage at 4 degrees C but was negative after 8 weeks. All samples of cheddar and Italian cheeses, most of which showed a positive phosphatase test, were negative for Y. enterocolitica. One out of 265 samples (0.4%) of pasteurized fluid dairy products contained Y. enterocolitica.


Reversible pulmonary oxygen toxicity in the primate. In order to investigate the effects of high concentrations of oxygen on the lung, experiments were performed on 18 baboons exposed to a humidified environment of 95% oxygen for five days. Open lung biopsies for biochemical assay, histologic and electron microscopic analysis and measurement of tissue respiration were performed before and after oxygen exposure. Pulmonary function was evaluated by measurement of arterial blood gases, compliance, closing capacity (CC), functional residual capacity (FRC), total lung capacity (TLC), residual volume (RV) and vital capacity (VC) before and after exposure and then at seven and 14 days in the animals which recovered. Six baboons removed from the oxygen environment after 96--110 hours and exposed to room air died within three to 20 hours of profound hypoxemia (PaO2 40 +/- 6). The remaining 12 baboons were successfully weaned to room air over a three day period with a return of ABGs to control values (PaO2 89+/- 2). Electron microscopic analysis of alveolar membranes exposed to 120 hours of hyperoxia demonstrated endothelial cell swelling, interstitial alveolar membrane edema, and an increased predominance of Type II pneumocytes. Lung volume measurements showed significant decreases in TLC (25%), VC (34%), CC/TLC (28%) and dynamic compliance (47%). Biochemical studies indicated a shift toward anaerobic metabolism with a decrese in tissue oxygen consumption, reduced cytochrome oxidase activity, and increased lung lactic acid production. These changes were all found to be reversible in the 12 baboons slowly weaned back to room air.


[Ultramicroscopic structure of the cyst wall of Entamoeba invadens, E. histolytica and E. coli]. The ultrastructure of the wall of E. invadens cysts induced in axenic cultures was compared by transmission electron microscopy of thin sections, with that of E. histolytica and E. coli cysts obtained from feces of human asymptomatic carriers. The cyst wall of the three species studied had similar ultrastructural features; in all, the wall was composed mainly by the aggregation of filaments adjacent to the plasma membrane. The cytoplasmic components which might be involved in the synthesis and deposit of the cyst wall of E. invadens appear to be different to those of E. coli and E. histolytica cysts.


[Instability of the DNA content in Entamoeba histolytica]. DNA content was repeatedly determined during a 10-month period on axenically cultured tropho-zoites from three strains (HK-9:NIH, HM-2:IMSS and HM-3:IMSS) and a clone of HK-9:NIH strain of Entamoeba histolytica, by means of the diphenylamine reaction. Average DNA content was more or less variable in all of them. According to their extent, variations were: 1) large (almost four times in strain HM-2:IMSS), or 2) small (1.5 times in the clonal isolate). With this relatively small sample, three statistically different groups were formed with nuclear DNA averages (1.35, 2.05 and 7.78 picograms respectively). Changes of ploidy and/or genomic heterogeneity in trophozoites could be the main causes of the striking instability in their DNA content.


[Amino acid composition in total proteins of strains of Entamoeba histolytica in axenic cultures]. In this paper we detail the amino acid composition of total protein hydrolyzates from trophozoites of two Entamoeba histolytica strains (HK-9:NIH and HM-2:IMSS) cultured axenically in TP-S-1 medium. Since we obtained nearly identical results in both strains, amino acid patterns may constitute an additional taxonomic criterion of the species. Our data are strikingly different from those found previously by other workers with total proteins from HK-9:NIH trophozoites cultured in a bacteria-conditioned medium; such difference emphasize the importance of axenic culture for the precise study of the chemical composition and metabolism of E. histolytica. Amino acid composition of total trophozoite proteins could be of use to attempt the design of a culture medium of defined amino acid content.


[Pulmonary thromboembolism. Occlusion of the right branch of the pulmonary artery caused by lung cancer]. A rare case of an oat cell carcinoma is presented. The clinical picture were those of pulmonary thromboembolism and other extrapulmonary symptoms. There were not respiratory symptoms directly related to the lung carcinoma and the chest film remain normal.


Uptake of selected pyrimidines by axenically grown Entamoeba histolytica. This study was conducted to determine the mode(s) of uptake of selected pyrimidine bases and nucleosides by axenically grown Entamoeba histolytica. Two-minute uptake studies with 3H-labeled compounds showed the following: 1) uridine and cytidine are taken up, in part, by a carrier mediated system; 2) uracil, cytosine, thymine and thymidine enter amebae via passive diffusion; both cytidine and uridine are taken up by passive diffusion when present at exogenous concentration greater than 500 muM; 3) specific carrier sites are used for transport of uridine-cytidine and uridine-adenosine; 4) cytidine and uridine uptake is markedly inhibited by iodoacetate and N-ethylmalemide; 5) hypoxanthine (10mM) stimulates uptake of uridine; 6) ribose fails to inhibit uptake of cytidine and uridine; 7) cytidine uptake exceeds that of uridine (i.e., apparent Vmax for: a) cytidine = 10.0 nmoles/2 min/10(6) amebae; b) uridine = 3.0 nmoles/2 min/10(6) amebae); 8) Kt for: cytidine = 0.30 mM; uridine = 0.21 mM; 9) Q 10's at 35.5 C vs 25.5 C for: a) cytidine = 3.9; b) uridine = 2.6.


[Immunochemistry of immunogenic proteins of Entamoeba histolytica]. Entamoeba histolytica (HK-9: IMSS) trophozoites were separated from the culture medium and washed and submitted to the effects of ultrasound during 10 minutes. After 10,000 rpm x 30 centrifugation, the supernatant fluid was separated in four fractions by Sephadex G-200 gel chromatography. These fractions were lyophilized, redisolved in a lesser volume and dyalized against isotonic saline solution. Antigen-antibody precipitant reactions between each of them and two rabbit antiserums were demonstrated: anti amoeba and anti fraction A. It was verified that the antisera reacted also with the culture medium, the human seric albumin and the sera from health people and patients with invasive amebiasis. The higher antigenicity, quantitatively and qualitatively, was obtained with the fraction B; the less antigenic fraction was the fraction D. In fraction A three antigens were identified. When those proteins were filtered by a Bio Gel P4 column, two fractions were eluted: A1 and A2 which were lyophilized, redisolved and dyalized in isotonic saline solution. After this treatment, fraction A1 was capable to maintain its antigenicity against the rabbit antisera and the sera from patients with invasive amebiasis, fact by which it was considered a protein from amebal origin and not as a protenic contaminant from the culture medium.


[Cytopathogenic activity of extracts E. histolytica trophozoites]. Previous studies have not shown any correlation between enzyme activities and pathogenicity of Entamoeba histolytica. The activities analyzed were selected without having proved their involvement in the cytopathogenicity. We are using the reverse approach to study the cytopathogenic factors, i.e., to detect and characterize first the cytopathic effect of amebic extracts on animal cells, and subsequently to try to identify the components involved and to explore their correlation with amebic virulence. Total extracts from E. histolytica trophozoites exert dosedependent toxic and lytic effects upon an animal cell line in culture. Erythocytes from eight mammaliam species have been used as a simple target model for the characterization of the cytolytic affect. Hemolysis by amebic extracts is consistent, reproducible, strain specific and dosedependent. Hemolytic potency is much larger E. histolytica extracts than those from other Entamoeba species. This work appears to open up the study of the biochemical basis of E. histolytica cytopathogenicity.


[Effects of nordihydroguairetic acid and ethanol on the growth of Entamoeba invadens]. The effects of the alcoholic extract (resin) obtained from Larrea tridentata, of nordihydroguiaretic acid (NDGA) and of ethanol over axenic cultures of Entamoeba invadens PZ strain were studied. The activity of these substances was demonstrated in growht curves. Inocula of 1-3 x 10(5) trophozites by milliliter in culture tubes containing 11 ml of TP-S-1 medium were done, and incubated at 25 degree C. In each experiment the follwoing was done: a) tipical growth curve as a control, b) curve with the medium plus 0.2 ml of a mixture of ethanol and water v/v  (solvent used for resin and NDGA), and c) curves with different concentrations of the test compounds. The trophozoite number was counted each 24 hours by means of an hemocitometer. The results obtained indicate that the resin has inhibitory activity at a concentration of 0.0001 per  cent. The NDGA activity was observed at 10(-6) to 10(-8) M concentrations. The ethanol had, "per se", inhibitory effect over the cultures, and was annulated by the  presence of NDGA. At higher contentrations of the acid acid the temporal curse of the graphas was similar to that observed for the control.


The development and standaridization of an ELISA method for the detection of Entamoeba histolytica antigens in fecal samples. Recognizing the difficluties associated with the microscopic detection and identification of Entamoeba histolytica in fecal samples, an immunochemical method of detection of E. histolytica antigens in fe-es has been developed. For a number of reasons, e.g., the known fragility of E. histolytica trophozoites, it was felt that intestinal amebiasis would be associated with detectable levels of E. histolytica anti-ens in feces from infected patients. A technique for harvesting amebic antigens in fecal samples was developed. An enzyme labeled antibody technique was used to visualize the harvested antigens. The inherent problems of performing any immunochemical assay in fecal samples have been addressed. The enzyme labeled antibody method is simple and is designed to be more sensitive than microscopic stool examination methods. Preliminary results indicate that the method is specific and sensitive.


[Identification of Entamoeba histolytica membrane antigens with antibodies of amebiasis patients]. The specificity of sera from patients with amoebiasis was assayed on trophozoites of E. histolytica HK9, HM15 and HM2 cultured axenically and monoxenically. Glutaraldehyde fixed cell were incubated with samples of serum, the excess of Igs was washed out and the presence of unbound material of specifically bound antibodies was measured with radiolabeled protein A from Staphylococcus aureus. A positive reaction with HK9 was observed in 18 per cent of the 66 sera tested. An immune crossreaction resulted with HM2 and HM15, while no reaction was observed with the nonpathogenic strains E. invadens, E. moshkovskii and E. laredo.


[Phagocytosis of human erythrocytes by Entamoeba histolytica. Quantitative study]. The degree of eythrophagocytosis of two recently isolated strains of E. histolytica was measured by microscopic examination. Amebas isolated from a patient with amebic rectocolitis (strain HM22:IMSS, monoxenic) ingested human red blood cells faster and in larger numbers than trophozoites isolated from an asymptomatic carrier (HM27:IMSS, monoxenic). The results suggest that an increased rate of phagocytosis could be one of the surface properties characteristic of the invasive strains of E. histolytica.


Production of cecal lesions in newborn guinea pigs with axenically cultivated Entamoeba histolytica. The production of cecal lesions with axenically cultivated Entamoeba histolytica in an experimental animal is reported for the first time. Forty-three newborn guinea pigs (one to five days old) were inoculated via laparatomy with 2.5 x 10(5) to 2 x 10(6) strain HM-1-IMSS amebae. Of 23 animals surviving to the 7th postinoculation day all showed cecal lesions at necropsy regardless of the age of the animal when used or the number of amebae received. Ten animals had lesions visible to the unaided eye and 13 had only microscopic lesions. No lesions were noted in other organs of the abdominal or thoracic regiosn. The use of axenically cultivated ameabae in newborn guinea pigs should prove a useful experimental model for the study of amebiasis.


[Culture and establishment of new Mexican Strains of Entamoeba histolytica]. Mixed and monoxenic cultures of rectal exudates from three patients with amebiasis of colon, and of feces from three asymptomatic carriers of Entamoeba histolytica obtained after administration of a saline cathartic, and from a case of exudate of the perianal cutaneous lesion border produced by this parasite, were done. The culture media used were for: mixed culture, Boeck and Drbohlav modified culture and, for monoxenic culture, Diamond's monophasic TP-S-1 medium without vitamin mixture and using bovine serum. In order to inhibit the bacterial growth in the mixed culture, streptomycin sulphate and pencillin G were used. In order to eliminate the bacterial flora kanamicin sulphate, chloramphenicol, gentamycin and disodic carbenicillin were added. Mixed cultures of seven strains of Entamoeba histolytica were obtained: HM22:IMSS, HM23:IMSS, HM24:IMSS, HM26:IMSS, HM27:IMSS, HM28:IMSS y HM29:IMSS, and monoxenic culture with Bacteroids symbiosus of the strains HM22:IMSS and HM27:IMSS, with a previous adaptation in mixed culture. The adaptation of HM26:IMSS strain in monoxenic culture was obtained by means of direct seeding of the rectal exudate with Fusobacterium symbiosus.


Cytotoxicity of a cell free extract of Entamoeba histolytica. Previous attempts to demonstrate toxicity of cell-free extracts prepared from axenically cultivated E. histolytica have proved unsuccessful. We report here the successful production of cytopathic effect by cell-free extracts of two strains (HM-1 and HK-9) of E. histolytica in tissue culture monolayers. The cytotoxic effect demonstrated was sensitive to serum in the assay medium. Failure of past investigations to demonstrate cytotoxic activity may be explained by the presence of serum in the assay systems used.


Clonal growth of Entamoeba in agar: some applications of this technique to the study of their cell biology. A new technique for growing single Entamoeba trophozoites into colonies in agar has been developed. This method depended upon axenic cultivation and utilized Diamond's new TYI-S-33 medium. The present paper describes several different types of experiment which have utilized the agar technique. a) Cloning. The isolation of clones from agar way easy and successful. Clones of the HM-1 strain did not differ in virulence for newborn hamster liver or in colony morphology or colony forming efficiency. b) Viability measurements. Colony forming efficiency (CFE) was proportional to the number of viable cells in a culture. Pilot studies with the amebacides, metronidazole and emetine, showed that the agar method should be useful in drug testing. Colchicine at high concentration inhibited clonal growth in a non-specific manner. c) E. histolytica HM-1 and E. invadens cells were rapidly killed by exposure to 42 degree C but survived relatively well at 0 degree C. d) Hemolysis. In a preliminary experiment HM-1 colonies did not produce halo's of hemolysis when grown in agar containing sheep red blood cells.


Restoration of virulence of axenically-cultivated Entamoeba histolytica by cholesterol. The lost pathogenicity of two strains of Entamoeba histolytica, one isolated in 1924 and the other in 1967, grown in axenic culture for the past five and six years respectively, was restored by supplementing the culture medium with cholesterol through a number of transfers. The number of passages in the cholesterol supplemented medium, necessary to restore a certain degree of pathogenicity of the two strains in hamsters, was proportional to the total time of in vitro cultivation of the strain, and not just the time of cultivation under axenic conditions. Pathogenicity, once restored, persisted for a long time after cholesterol treatment was stopped.


[An in vitro model for the quantitative study of E. histolytica virulence]. An experimental model for the quantitative study of the cytopathic effect of E. histolytica is described. Confluent cultures of the MDCK cell line, which form a monolayer with morphological and electrophysiological features of an epithelium were used. Trophozoites of the strains HM1:IMSS, HM2:IMSS and HM3:IMSS, axenic produced variable degrees of cytopathic effects of MDCK cultures each one characteristic for a given strain. The degree of damage induced by the different strains of amebas was measured quantitatively. The results demonstrate that the degree of the cytopathic effect produced in vitro by the various strains tested correlates with the virulence shown in vivo by the same strains, judged as the size of the inoculum required to induce liver cytopathic effect in newborn hamsters.


[Interaction between Entamoeba histolytica and the cecal epithelium in the guinea pig]. The early interaction between  E. histolytica trophozoites and the intestinal mucosa was studied by light and electron microscopy using sheets of isolated epithelium of the guinea pig colonic mucosa. The integrity of the isolated epithelium was confirmed by scanning electron microscopy. After 30 minutes of interaction, the amebas adhered first, and penetrated subsecuently the epithelium of the colon at the interglandular regions. The results suggest that the trophozoites of E. histolytica invade the epithelial barrier exclusively at the regions of lowest resistence, represented by the interglandular zones of epithelial shedding.


[Ultramicroscopic study of the invasion of the mucosa of the colon by Entamoeba histolytica in axenic cultures]. Animal models have been utilized to study experimentally produced intestinal amebiasis. The guinea pig has been the animal of choice to study the mechanisms by which trophozoites of E. histolytica produce colonic lesions. The purpose of this study was to produce lesions in the colon of newborn guinea pigs by the injection of E. histolytica, HM-1 IMSS, strain grown axenically, and to characterize the early changes with the light and electron microscopes. A total of 35 guinea pigs, one to two days old, were inoculated into the cecum with one and a half million of trophozoites of E. histolytica. Thirteen surviving animals were sacrificed. In four there were clear cut amebic lesions. Sections were taken from neighbouring transitional zones. The most important changes were the migrations of trophozoites to the bottom of colonic glands with many parasites located within the crypts. There was passage of ameabae to the lamina propia with practically no inflammatory reaction. It is thought that the parasite, in contact with epithelial crypt cell, produce direct lysis of these elements leading to migration of amebae into the lamina propia.


[Serology with polysaccharides of Entamoeba histolytica]. The immunochemical activity of a polysaccharidic antigen derivated from E. histolytica was studied. This polysaccharide was tested by means of counterimmunoelectrophoresis (CIE) and radial immunodiffusion (IDR) against sera from patients with diagnosis of intestinal and hepatic amebiasis. In order to perform a test most significative to the problem sera, these were submitted previously to passive hemagglutination, CIE and IDR with total ameba antigen. In hepatic amebiasis it can be observed that the CIE done with commercial antigen is practically negative in all cases; incontrast, the test performed with total antigen was positive in almost all the cases. Passive hemagglutination with sheep red blood cells, tanned and sensibilized with total ameba antigen gave very high titers when the problem was amebic hepatic abscess, and very low when the problem was intestinal amebiasis. Counterimmunoelectrophoresis and radial diffusion were negative with polysaccharidic antigen in all the studied sera.


[Activity of the alternative complement pathway by various axenic strains of Entamoeba histolytica]. The activation of the alternative pathway of complement by axenic cultures of E. histolytica is reported. The activation is carried out in different proportions by several axenic strains, i.e., HK9:NIH, HM1:IMSS, HM2:IMSS and HM3:IMSS; the end result consists of a destruction of the parasite as well as consumption of the serum complement. The participation of the alternative pathway was studied in the presence of EGTA and Mg++, zymosan, heat-inactivation at 50 degree C for 20 min and immunoelectrophoretic studied with anti-factor B.


[Immunite reaction of hamsters infected or immunized with Entamoeba histolytica]. The immune response in hamsters either infected or immunized with Entamoeba histolytica was studied. The response was tested by determination of antibodies by the counterimmunoelectrophoresis technique and by lymphocyte transformation induced by the amebic antigen. The results obtained showed differences among both groups when the ability of amebic antigen to induce 3H-thymidine incorporation of lymphocyte cultures is compared. However, the response of lymphocyte cultures to Concanavalin A was similar in both studied groups. On the other hand, a group of hamsters hyperimmunized with E. histolytica antigen incorporated in complete Frend's adjuvant showed a greater immune response than the group receiving a single dose.


[A new hemagglutination method to determine antibodies against Entamoeba histolytica]. A passive hemagglutination method useful for the determination of antibodies against Entamoeba histolytica is described. The E. histolytica antigen is attached to group "O" human red blood cells by means of CrCl3. The specificity and sensitivity of the CrCl3 method was compared with other hemagglutination methods, i.e., tannic acid method and a commercial preparation, as well as counterimmunoelectrophoresis. The CrCl3 method showed a reactivity similar to the tannic acid hemagglutination technique and greater specificity than the commercial preparation. The assay reported here showed a good correlation with the counterimmunoelectrophoresis method. The sensitization of erythrocytes with the amebic antigen by means of CrCl3 offers an easy, rapid and specific technique which can be used as a routine test for the detection of antibodies against E. histolytica.


Effect of carbon source on enzymes involved in glycerol metabolism in Neurospora crassa. Specific activities of eight enzymes involved in glycerol metabolism were determined in crude extracts of three strains of Neurospora crassa after growth on six different carbon sources. One of the strains was wild type, which grew poorly on glycerol as sole carbon source; the other two were mutant strains which were efficient glycerol utilizers. A possible basis for this greater efficiency of glycerol utilization was catabolite repression of glyceraldehyde kinase by glycerol in wild type, and two-fold higher glycerate kinase activity in the mutant strains after growth on glycerol, thus apparently allowing two routes for glyceraldehyde to enter the glycolytic pathway in the mutant strains but only one in wild type. The preferential entry of glyceraldehyde to the glycolytic pathway through glycerate was suggested by the lack of glyceraldehyde kinase in all three strains after growth on one or more of the carbon sources and the generally higher levels of aldehyde dehydrogenase and of glycerate kinase than of glyceraldehyde kinase.


Cyclic AMP and cyclic GMP in germinating conidia of Neurospora crassa. A new method for obtaining synchronous germination allowed accurate time-course studies of endogenous levels of cyclic adenosine 3',5'-monophosphate and cyclic guanosine 3',5'-phosphate in germinating conidia of Neurospora crassa. The levels of both cyclic nucleotides remained constant throughout germination, showing that they neither signal nor respond to any of the biochemical changes which are known to occur in the germination process. Conidal germination was approximately normal in three cr-1 mutants of Neurospora which have been shown to be deficient in adenylate cyclase activity. Cyclic AMP levels in the mycelia of cr-1 mutants were low, but surprisingly, the levels in conidia were normal.


Effect of temperature on diauxic growth with glucose and organic acids in Pseudomonas fluorescens. Growth of Pseudomonas fluorescens in batch culture with glucose and organic acids resulted in typical diauxic responses at 30 degrees C but no detectable diauxic lag at 5 degrees C. At 30 degrees C, organic acids were preferentially utilized during the first growth phase. Glucose utilization was delayed until onset of the second growth phase. Systems involved in direct uptake and catabolism of glucose responded in a manner compatible with repression by malate and/or its metabolites and induction by glucose and/or its metabolites. The oxidative non-phosphorylated pathway, through gluconate and 2-ketogluconate (2-KG) as intermediates, was not induced during either growth phase. At 5 degrees C, growth with glucose and organic acids was biphasic but without diauxic lag. Organic acids were preferentially utilized during the first growth phase. Although carbon from glucose was not fully catabolized until onset of the second growth phase, glucose was oxidized to and accumulated extracellularly as gluconate and 2-KG during the first growth phase. No significant repression of glucose-catabolizing enzymes was observed during growth with organic acids in the presence of glucose. However, uptake activities for gluconate and 2-KG did not increase significantly until onset of the second growth phase. Thus, at low temperatures, psychotrophic P. fluorescens oxidized glucose to extracellular 2-KG, while growing on preferred carbon sources. The 2-KG was then catabolized after depletion of the organic acid.


[Current status of immunoprophylaxis against foot-and-mouth disease]. An analysis is made of the latest developments and importance of immunoprophylaxis against FMD, with reference to the epizootiological situation of the disease and its bearings, today, on industrialized livestock production. Comprehensive immunoprophylactic action (vaccination of all cattle every year) has been taken first by the GDR and later on by more European countries, as well. As a result of such action the disastrous impact of FMD and its associated high loss rate have been considerably mitigated. A decisive role, in that context, has been played by the development of effective inactivated vaccines for swine. An FMD live vaccine has been devised in GDR for after-care of swine. It is not only harmless to the animal, but, as well known, it provides numerous advantages over adsorbate vaccines. FMD immunoprophylaxis for full success depends primarily on the proper selection of virus strains suitable for vaccine produktion, vaccine quality, avoidance of post-vaccination damage or disease, and the availability of an efficient concept for vaccine application. The above problems are expounded in this paper, and derived from them are substantive conclusions for further studies, with the view to improving FMD immunoprophylaxis.


[Results of veterinary tumor research and their significance for practice]. Enzootic bovine leucosis and Marek's disease of fowl are the most important tumour diseases of farm animals, with the greatest repercussions on the national economy, today. Experimental findings obtained from either disease in recent years are reported in this paper, and an assessment is made of their importance to practice. In the context of enzootic bovine leucosis, particular reference is made to those findings which helped to establish the presence of virus infection, results by which the pathogen could be characterised, and to studies undertaken with the view to devising methods for virologico-serological diagnosis. In the context of Marek's disease, reference is made to virological, epizootiological, and diagnostic aspects, however, with emphasis being laid on possibilities for immunisation and, in that connection, on known fundamental concepts for successful immunoprophylaxis against that tumour disease.


[Transmissible gastroenteritis of swine as a model for infectious diarrhea]. Transmissible gastro-enteritis is a virus-caused diarrhoeal disease of swine which may result in up to a total loss among newborn animals. Changes are, primarily, of pathologicoanatomic nature and take place in the small intestine. The epithelium of the intestinal villi is completely destroyed, and diarrhoea is the result. Deformation in blood composition is a secondary sequel. The diarrhoea proper causes dehydration and acidosis which, in conjunction with abnormal heart function due to hyperkalaemia, cause death of the piglets affected. Reference is made to the intrinsic immunological conditions in swine, and possibilities are discussed for immunoprophylactic control of transmissible gastro-enteritis by induction of effective lactogenic immunity. The approaches taken to vaccine application should largely correspond to the natural route of infection.


Drug and family therapy in the aftercare of acute schizophrenics. After a brief inpatient hospitalization, 104 acute, young schizophrenics, stratified by premorbid adjustment, were randomly assigned to one of four aftercare conditions for a six-week controlled trial. Conditions involved one of two dose levels of fluphenazine enanthate (1 ml or 0.25 ml) and presence or absence of crisis-oriented family therapy. Relapses during the six-week period and at six-month follow-up were least in patients who received both high-dose and family therapy (0%) and greatest (48%) in the low-dose-no therapy group. Brief Psychiatric Rating Scale symptom ratings disclosed a significant family therapy effect at six weeks that was sustained at six months only for therapy patients originally receiving the high drug dose. Numerous interactions were found between premorbid adjustment status and response to the two treatment conditions.


The effect of physostigmine on normal human sleep and dreaming. Physostigmine, an anticholinesterase that increases the action of brain acetylcholine, induces rapid eye movement (REM) sleep in normal humans. In this study we show that man dreams during physostigmine-induced REM sleep. Seventeen normal volunteers were pretreated with methscopolamine and received one intravenous infusion per night of either placebo or physostigmine either ten or 35 minutes after sleep onset. Subjects were awakened at specific times after infusion and interviewed regarding any sleep mentation prior to awakening. Results indicated that dreaming occurred during physostigmine-induced REM periods but that physostigmine did not alter mentation during non-REM sleep. These dreams were similar to spontaneous REM sleep dreams in content, vividness, unusualness, and emotionality.


Nocturnal sleep in separated monkey infants. Nocturnal sleep was recorded from ten unrestrained, group-living Macaca nemestrina (pigtail) monkey infants, using implantable multichannel biotelemetry systems, during the agitation-depression behavioral reaction that follows maternal separation. Sleep disturbances during the four nights of separation were characterized by decreases in rapid eye movement (REM) time and in the number of REM periods, and increases in REM latency. Time awake and number of arousals were increased. Slow-wave sleep was not significantly affected. Sleep pattern changes were most pronounced the first separation night, and tended to decrease as separation continued, whereas behavioral measures of depression tended to increase as separation continued (up to four days). Sleep patterns returned to normal following reunion with the mother. Those infants who had the most severe sleep disturbances the first separation night (more time awake, less total sleep, less REM) also tended to become most depressed behaviorally later in the separation period.


Murine leukemia virus in chemically-induced sarcomas as detected by XC plaque formation. To detect whether methylcholanthrene induced murine sarcomas contain endogenous murine leukemia viruses, direct XC syncytium assays were performed over short term tumor cultures. Out of 31 sarcomas, 13 (42%) induced more than 4 plaques per flask, while no such activity was found in 39 control spleen samples. It is postulated that the positive tests are related to an increase in the titer of endogenous B-tropic virus, probably as a result rather than as a cause of tumor induction by chemicals.


Withdrawal from long-term high-dose desipramine therapy. Clinical and biological changes. Investigation was undertaken on a patient whose long-term intake of desipramine hydrochloride was amongst the highest reported. Desipramine treatment instituted at a daily dosage of 75 mg for depressive equivalents of head, chest, and abdominal pain was increased to 1,000 mg daily over a 12-year interval with minimal side effects. Plasma desipramine level dropped immediately on withdrawal, and urinary metabolite values dropped over the subsequent five days. The electrocardiographic abnormalities of first-degree atrioventricular block and incomplete left bundle branch block rapidly disappeared on cessation of medication. Electroencephalographic changes with symmetrical generalized irregular 5- to 7-cps theta activity and 18- to 28-cps beta activity also improved. Longitudinal polygraphic sleep studies showed prolonged rapid eye movement rebound and increased delta sleep coincident with withdrawal. It took ten days after cessation of desipramine for urinary 3-methoxy-4-hydroxyphenylglycol concentration to increase substantially. Although catecholamines are involved in growth hormone (GH) and cortisol regulation, no abnormalities were found in GH or cortisol levels.


In vitro growth characteristics and heterogeneity of mouse hepatitis virus type 3. The in vitro virus yield of MHV3 reached 10(7) PFU/ml in mouse DBT cells infected with a virus suspension in HEPES-buffered medium containing DEAE-dextran. The virus titer was 10(6) PFU/ml in the presence of 10 micrograms actinomycin D/ml MHV3 growth in DBT cells gave three peaks of density (1.10--1.14 g/cm3, 1.18--1.20 g/cm3, and 1.25--1.31 g/cm3) in sucrose gradients. All these peaks retained infectivity.


Differentiation between specific and nonspecific reactions of bovine sera and foot and mouth disease virus (FMDV) in immunodiffusion tests. The precipitating and neutralizing activities of normal bovine sera with FMDV were studied and compared. Twenty-two out of 79 normal bovine sera gave a positive reaction in micro neutralization tests with FMDV type O, while six did so with type A. In RID tests 32 sera were positive with type O and 28 with type A virus. Almost all of the 79 sera gave a positive reaction in the RID with trypsin treated virus of both types. After three to four fold concentration most sera also gave visible reactions in ID tests when tested against complete virus. When O virus was used the ID patterns produced by most normal sera clearly differed from those obtained with early and late convalescent sera from FMDV infected steers. When type A materials were employed this was also the case but to a lesser extent. The patterns obtained with concentrated normal sera showed, in general a strong line with trypsin treated virus and no line or a weaker one with complete virus. The substances in normal bovine sera precipitating trypsin treated O virus were different from those reacting with trypsinized virus of type A.


Polyethylene glycol-mediated infection with avian sarcoma viruses. In this communication we report that fusion of viral and cellular membranes by polyethylene glycol is a convenient approach to overcoming genetically determined resistance to infection. Using this method, avian sarcoma virus-transformed mammalian cells have been produced which serve as useful model systems for the study of Rous sarcoma virus-specific tumor antigens.


The structure of the Rous sarcoma virus glycoprotein complex. The viral envelope glycoprotein gp85 is released from purified Rous sarcoma virus by treatment with 2-mercaptoethanol, while the second surface antigen gp35 remains associated with the membrane of intact virus particles. The data represented substantiate and extend previous observations on the Rous sarcoma virus glycoprotein complex.


In vitro RNA synthesis in nuclei from herpes simplex virus infected cells: effects of ultraviolet-irradiated, defective and bromodeoxyuridine-resistant virus preparations. Transcription of virus DNA was detectable in vitro in nuclei isolated from cells infected with wild type and bromodeoxyuridine (BUdR)-resistant herpes simplex virus type 1 but was suppressed in nuclei of cells infected with ultraviolet irradiated or defective virus or cells treated with BUdR during infection.


Various heterologous cells exhibit interferon induced transfer of viral resistance. Previously it was shown that cocultivation of mouse L and human WISH or baby hamster kidney cells in the presence of mouse interferon resulted in decreased viral yield from both cell species. We now show that this phenomenon also occurs when rabbit kidney and human WISH cells, with their corresponding interferons, are cocultivated with human WISH and baby hamster kidney cells, respectively. This finding increases the number of donor cell types to three. The related finding that monkey VERO and chick embryo cells can be recipients of transferred resistance expands the number of heterologous recipient cell species capable of receiving transferred resistence to five. Not all cell types tested have been shown to function in this transfer system. The fact that VERO cells, which do not produce interferon, are capable of receiving transferred resistence is significant because it indicates that the mechanism of transfer does not involve production or interferon by the recipient cells.


Disulfiram neuropathy.

Disulfiram neuropathy. A 35-year-old man developed a distal sensorimotor polyneuropathy after taking disulfiram, 500 mg daily for five months. His symptoms improved after the drug therapy was discontinued. Clinical, electrophysiological, and pathological observations during the acute stage and during recovery suggest that disulfiram produces a distal axonopathy.


Herpes simplex virus subunit antibodies in patients with Parkinson's disease. Serum IgG antibodies against herpes simplex virus (HSV) type 1 capsid, envelope, and excreted antigens in 52 patients with idiopathic Parkinson's disease, and in their age- and sex-matched controls, were assayed with a solid-phase radioimmunoassay. When compared with the controls, patients with Parkinson's disease were found to have a substantially increased antibody response against each of the HSV subunit antigens tested. The increased antibody response in patients with Parkinson's disease was not associated with the occurrence of recurrent HSV infections, since the difference in antibody levels was most evident when comparing patients without recurrent HSV infections with their respective control group. Consequently, the increased HSV antibody response in patient with Parkinson's disease might depend on some antigenic stimulation other than ordinary recurrent HSV infections, or alternatively, on the generally enhanced immunological reaction of the patients against HSV.


Serum pyridoxal concentrations in patients with diabetic neuropathy. Serum pyridoxal (vitamin B6) concentrations were measured in 50 patients with significant diabetic neuropathy. There were 24 males and 26 females with a mean age of 58.2 years and a mean duration of diabetes of 9.8 years. The level of pyridoxal was significantly lower in these patients when compared with randomly selected diabetic patients matched for age and sex without clinical evidence of neuropathy. There was no significant difference in the duration of the diabetes between the two groups. The results indicate an association between pyridoxal deficiency and neuropathy in diabetic patients.


Duration of vaccinal immunity against Marek's disease. Specific vaccinal immunity against Marek's disease (MD) virus was measured by a short-term challenge test in which chickens vaccinated with turkey herpesvirus (HVT) and control chickens at various ages up to 40 weeks were inoculated with virulent MD virus and evaluated 2 weeks later for microscopic lesions and MD viremia. Vaccinal immunity induced by high (2,900 PFU) and low (29 PFU) doses of HVT persisted without detectable reduction through the 40th week in line 151 chickens, thus indicating that resistance of older vaccinated chickens to MD may not be based solely on general age resistance (immunocompetence) or on subsequent exposure to field MD virus. The HVT viremia titers (number of buffy-coat cells inducing virus plaques in vitro) decreased with increasing age, but even chickens from which no HVT was isolated were usually immune to MD. The dose of vaccine appeared to influence, at least slightly, the magnitude of MD immunity and vaccine virus viremia.


Lymphoproliferative diseases of fowl: characterization of transplantable G-B1 Marek's disease tumor cells in culture. A transplantable Marek's disease tumor, derived and maintained through in vivo passage in syngeneic G-B1 chickens, was established in cell culture and characterized. The cells (GBT cells) grew in suspension and appeared morphologically similar to other Marek's lymphoblastoid lines except for prominent nucleoli in GBT cells. The number of chromosomes increased during in vitro cultivation from near triploid to subtetraploid. Chromosome no. 3 was abnormally short. The GBT cells retained surface B1 histocompatibility antigen of the original tumor as well as Marek's disease tumor-specific surface antigen (MATSA); however, infectious Marek's disease virus was not detected by immunofluorescence, electron microscopy, inoculation into susceptible birds, or assay on susceptible cells in culture. Inoculation of as few as 100 cultured cells into syngeneic G-B1 chickens results in tumor formation at the inoculation site, with eventual death of the recipients.


Differentiation of avian infectious bronchitis virus isolates by thermal sensitivity. Six isolates of avian infectious bronchitis virus (IBV) at different passage levels in the chicken embryo were tested for sensitivity at 56 C, with the following results: 1) isolates differed in thermal sensitivity; 2) virus was inactivated in a first-order exponential-kinetics 2-component fashion indicative of a heterogeneous population with a preponderance, 98% or more, of thermal-sensitive (S) virions over thermal-resistant (R) virons; 3) R virions 2-component populations were inactivated; 4) recovered R virions could be maintained in continuous passage in the chicken embryo as a one-component population by the limiting-dilution technique, although there was a progressive parallel increase in thermal sensitivity associated directly with the continuous passage; 5) growth curves of isolated one-component populations of R virions were quantitatively and in time sequence similar to curves of 2-component populations of continuous-passage virus.


Interaction of aflatoxin with infectious bursal disease virus infection in young chickens. Young White Leghorn chickens fed 2.5 microgram of aflatoxin (Afl) per g of diet from hatching until 4 weeks old and infected with infectious bursal disease virus (IBDV) at 3 weeks old had significantly higher mortality and more severely depressed body weights than chicks with aflatoxicosis or IBD alone. Afl-IBDV chicks also had more extensive gross and microscopic changes characteristic of IBD than did IBDV-chicks. None of the treatments significantly reduced antibody responses to Newcastle disease(ND) and infectious bronchitis vaccines or increased susceptibility to challenge with virulent NDV. In a similar experiment chickens fed Afl from hatching to 7 weeks of age had no marked depression in immune response to ND vaccination.


Experimental infection of turkeys with infectious bursal disease virus. Commercial turkey poults 3 to 6 weeks old were infected experimentally by eyedrop with an infectious bursal disease virus (IBDV) inoculum obtained from chickens experiencing clinical IBD. The IBDV was passed 6 successive times in poults in an attempt to increase its pathogenicity for turkeys. Regardless of passage level, the IBDV infection in poults was subclinical, with no morbidity, mortality, or gross lesions observed. The bursae of Fabricius from infected poults, however, displayed various degrees of microscopic degeneration and IBDV specific fluorescence at 3, 4, and 5 days postinfection (PI). Infected turkeys also developed low levels of virus-neutralizing antibodies against IBDV at 12 days PI. Uninoculated poults kept in the same unit with infected poults also displayed microscopic changes and IBDV specific fluorescence 7 days after their appearance in inoculated poults. The IBDV was isolated from infected poults only after 5 successive passages of bursal material from infected poults in 9-day-old chick embryos. The IBDV from infected embryos was inoculated into susceptible 3-week-old chickens and 5-week-old poults and produced IBDV fluorescence and microscopic pathology in the bursae of infected poults and clinical IBD in infected chickens.


Purification of avian encephalomyelitis virus by ultracentrifugation in a nonlinear cesium chloride gradient. A rapid and convenient method is described for purifying avian encephalomyelitis virus. Infected embryo homogenate was treated with polyethylene glycol followed by extraction with fluorocarbon. Extracted sample was pelleted at 73,400 x g, resuspended, and subjected to isodensity centrifugation in preformed nonlinear gradient. A distinctive virus band was observed after centrifugation for 120 min at 192,000 x g. The fraction containing the virus band demonstrated the major portion of infectivity and the presence of the virus particles by electron microscopy. The method was reproducible within a limited number of trials.


Avian adenoviruses: effect on egg production, shell quality, and feed consumption. Four avian adenovirus isolants (Indiana C, DPI-2, IBH-2, and B-3)were examined for pathogenicity in laying birds. None affected shell quality, and only one (IBH-2) had even a minimal effect on egg production (faster decline). Of three isolants tested (Indiana C, DPI-2, and IBH-2), only IBH-2 caused feed intake to be reduced. No other clinical signs were observed during these experiments. Egg transmission, virus excretion, and antibody production were found to mimic patterns reported for serotype-1 avian adenoviruses.