Gonadotropin releasing hormone release from the rat hypothalamus: dependence on membrane depolarization and calcium influx. Release of gonadotropin-releasing hormone (GnRH) was studied by incubating individual rat hypothalami for 60 min, after a 30-min preincubation period, and measuring GnRH in the medium by immunoassay. During the 1 h of incubation, endogenous GnRH release was linear and exogenous GnRH was not destroyed. Membrane depolarization produced by increasing the medium potassium concentration to 60 mM increased GnRH release to 200-500% of control. Membrane depolarization produced by adding 10(-5) or 10(-4) M ouabain increased GnRH release to 200% of control. Melatonin (10(-7) M) and prostaglandin E2 (4 X 10(-4) M) Aslo stimulated GnRH release to 200% and 170% of control, respectively. Inhibition of calcium influx by omission of medium calcium and addition of 0.05 M EDTA reduced GnRH release to 50% of control. Both no calcium-EDTA medium and verapamil (10(-5) M) prevented the stimulation of GnRH release by 60 mM potassium, 10(-3) M melatonin, and 4 X 10(-4) M prostaglandin E2. We conclude that hypothalamic GnRH release depends on membrane depolarization and calcium influx, as does the secretion of hormones from other endocrine tissues.


Comparison of inhibitory effects of 3,5,3'-triiodothyronine (T3), thyroxine (T4), 3,3,',5'-triiodothyronine (rT3), and 3,3'-diiodothyronine (T2) on thyrotropin-releasing hormone-induced release of thyrotropin in the rat in vitro. In order to compare, in vitro, the TSH suppressive effects of iodothyronines, rat pituitary quarters were first preincubated with T4, T3, rT3, or 3,3'-diiodothyronine (T2) in Gey and Gey buffer containing 1% bovine serum albumin for 2 h at 37 C and then incubated at 37 C for 1 h with the iodothyronine under study and TRH. TSH released into the medium during incubation was compared to that released by control pituitary fragments, which were not exposed to iodothyronines. All four iodothyronines (T3, T4, rT3, and T2) were able to significantly inhibit the TRH-induced release of TSH from pituitary fragments in a dose range of 0.015-2.2 microgram/ml. However, much larger doses of sodium iodide (1.25 mg/ml) and diiodotyrosine (10 and 30 microgram/ml) had no significant effect on the release of TSH. Among T3, rT3, and T4, T3 was the most potent and rT3 was the least potent. The relative potency of T3:T4:rT3 appeared to be approximately 100:12:1 when estimated from the lowest doses that caused significant inhibition of TRH-induced release of TSH, and approximately 100:6:0.5 when estimated from the doses that caused 50% inhibition of TSH release; the TSH inhibiting potency of T2 was similar to that of rT3. The activity of T4 could not be explained entirely on the basis of contamination of T4 with T3 or by in vitro conversion of T4 to T3. Similarly, the available data suggested that rT3 and T2 possess some, albeit modest, intrinsic TSH-Suppressive activity. TSH-inhibiting activities of T3, T4, and rT3 were also studied using pituitary fragments from starved and iodine-deficient rats. There was no evidence of a change in the sensitivity of the thyrotroph to either T3 or T4 in starvation. Similarly, comparison of the responses to several doses of rT3 did not indicate any significant abnormality in the sensitivity of the thyrotroph to rT3 in starvation or iodine deficiency. However, comparison of the TSH-suppressive effects of T4 in the iodine-deficient and normal rat indicated a significant increase in the sensitivity of the thyrotroph to T4 in iodine deficiency. A similar trend was also evident in the effect of T3 in iodine deficiency, but it fell short of statistical significance.


Participation of serotonin in thyrotropin release. I. Evidence for the action of serotonin on thyrotropin releasing hormone release. Injection of serotonin (5-HT) into the third ventricle of the rat resulted in a rapid increase of serum TSH; a significant effect was observed 5 min after injection, whereas the maximal effect appeared 10 min after the injection of 1 microgram 5-HT. This stimulating effect of 5-HT was completely prevented by pretreatment with cyproheptadine, a blocker of 5-HT receptors, whereas fluphenazine, a dopamine receptor blocker, was unable to block it. Third venticle injection of 5-HT in rats bearing anterior hypothalamic lesions (which did not affect the suprachiasmatic nucleus or the medio-basal hypothalamus) also induced an increase of serum TSH similar to that observed in normal rats despite the fact that these animals show a lower basal TSH. In vitro, the addition of 5-HT to an incubation medium containing one hemi-anterior pituitary did not modify medium TSH, whereas 5-HT addition induced an increase of medium TSH in the system containing one hemi-anterior pituitary and two hypothalami. We conclude that 5-HT acts on TSH function probably through a stimulation of TRH release.


Cellular heterogeneity in primary monolayer cultures of mouse pituitary thyrotropic tumors: separation of thyrotrophs. Primary monolayer cultures of cells derived by enzymatic dispersion from mouse pituitary thyrotropic tumors were demonstrated to be comprised of at least 2 cell types and were separated by selective culture techniques. Suspension cultures of spherical cells had the morphological characteristics of thyrotrophs and produced large quantities of TSH. Separated monolayer cultures of spreading, planar cells produced collagen but almost no TSH. These observations indicate that primary monolayer cultures of cells derived from thyrotropic tumors are heterogeneous, containing only a small proportion of thyrotrophs.


Acute stimulated hormone release from cultured GH3 pituitary cells. Treatment of cultured rat pituitary GH3 cells with 50 mM KCl in growth medium released 33% of cell PRL and 18% of cell GH with a half-time of 5 min. Hormone in the culture medium was increased 2- to 4-fold over unstimulated levels. The response required calcium; barium and strontium, but not magnesium, could substitute for calcium. Low temperature completely inhibited hormone release, which was also reduced significantly by inhibitors of energy metabolism and by nitrogen. This acute response was similar in ionic requirements, hormones released, and time course to the acute effect of TRH. Like potassium stimulation, TRH resulted in acute release of both PRL and GH. This contrasts with the finding that chronic TRH treatment reduced GH synthesis in GH3 cells. After a 10-min preincubation with potassium, subsequent short incubations with potassium released little hormone unless the cells were allowed to recover by incubation in normal medium for at least 2 h. This acutely releasable hormone pool seems to be located in a membrane-bound subcellular fraction, since GH3 cells did not discharge the cytoplasmic marker enzyme, lactate dehydrogenase, during potassium-stimulated hormone release.


The effect of L-DOPA administration on thyrotropin (TSH) and thyrotropin releasing hormone (TRH) levels in serum in primary or pituitary hypothyroidism. The effect of acute administration of L-DOPA on TSH and TRH levels in serum was studied in primary or pituitary hypothyroidism. TRH levels in serum fell and then returned to initial levels after L-DOPA administration in primary or pituitary hypothyroidism. TSH levels in serum fell and then returned to initial levels after L-DOPA administration in primary hypothyroidism. T4 and T3 levels in serum did not change after L-DOPA administration in primary or pituitary hypothyroidism. These data suggested that L-DOPA might act directly to hypothalamus.


Simultaneous competitive enzyme immunoassay for human chorionic gonadotropin. A simultaneous competitive enzyme immunoassay (SICEIA) for hCG was developed using beta-D-galactosidase (beta-Gal) as a labelled enzyme and anti-hCG antibody coated sheep red blood cells (SRBC) as a solid phase. In this report, a new coupling agent, MCAE, was used to couple beta-Gal with hCG. The sensitivity was improved to the degree of 2.5 mIU/ml, equal to that of RIA. The present procedure was safer and rapider than RIA. The value of hCG in urine by our procedure had good correlation with that by RIA.


Chemical sterilization of males - successful inhibition of spermatogenesis in langurs (Presbytis entellus entellus dufresne) after metopiron (SU-4885, Ciba) administration. 1. Daily administration of metopiron (SU-4885, Ciba) (10 mg/kg body wt. for 30 days) produced a massive atrophy of the spermatogenic elements. The pachytene primary spermatocytes have been implicated as the main site of damage. Seminiferous and Leydig cell shrinkage was conspicuous. Epididymal and vesicula seminalis structure and functions were severely affected. 2. RNA, protein and sialic acid contents of the testes, epididymides and seminal vesicles were reduced, whereas testicular cholesterol and enzyme phosphatase were elevated. 3. Marked reduction in the testicular glycogen was noticed. 4. Normal functioning of liver, kidney and general metabolic activities was revealed by clinical parameters (serum, transaminases, phosphatases, lipids, serum bilirubin, blood sugar and blood urea). Serum protein levels were low, whereas the serum cholesterol was elevated. 5. Metopiron (SU-4885, Ciba) at 10 mg/kg body wt. dose level did not cause severe damage to the vital organs but it produced an effective inhibition of spermatogenesis in male Presbytis in 30 days and thus induces an antifertility state.


[Comparison between a shortened TRH-TSH test and thyroid suppression test in the diagnosis of hyperthyroidosis]. In a selected group of fifty-one persons suspected to be hyperthyroid the comparison between a shortened variant of TRH-TSH stimulation test and thyroid suppression test was made. In patients whose other functional tests gave no univocal results, the agreement between suppressibility of radioiodine uptake and TSH response in 90,2% of cases could be established. High degree of correlation between both parameters (coefficient V = 0.791) permits to regard these tests as practically equivaluablein giving evidence of dysregulation between pituitary and thyroid gland. In persons with TSH-dependent thyroid function, however, no correlation between numerical values of radioiodine uptake and TSH concentrations after stimulations was found.


Efficacy and toxicity of the solvent polyethylene glycol 400 in monkey model. Several antiepileptic drugs, such as carbamazepine and clonazepam, have low bioavailability in solid form and are insoluble in an aqueous solution. Alcohol solvents are often employed as vehicles when these drugs are studied in animal models. Secondary and particularly tertiary alcohols are suspected of some anticonvulsant activity. The present research evaluated the possibility that polyethylene glycol 400 (PEG 400) might be efficacious, toxic, or both. Monkeys (N = 11) rendered epileptic by aluminum-hydroxide were administered PEG 400 by constant rate (1 ml/hr) intravenous infusion for 3--4 weeks, preceded and followed by several weeks of baseline. At a concentration of 60%, PEG 400 significantly reduced seizure frequency, but also exhibited severe side effects. These findings suggest that experimental testing of anticonvulsants may be compromised when this or similar solvents are used chronically.


The relation between insulin and adipocyte insulin receptors during treatment of human obesity. Seventeen obese patients were submitted to 1 week's severe then 3 months moderate dietary restriction. The glycaemic and insulinaemic response to oral glucose was measured at baseline, after 1 week and at 3 months, and the results related to adipocyte insulin receptors estimated from fat biopsies taken on each occasion and to the weight lost. Adipocyte insulin binding was found to be significantly inversely correlated with the insulinaemic response to oral glucose (r = -0.54, P less than 0.05), an inverse relationship maintained even after weight loss had been achieved and displayed within as well as between subjects. Variation of 125I-insulin binding to adipocytes was related to receptor number and not to receptor affinity and could not be accounted for by differences in 125I-insulin degradation. Adipose cell size did not correlate significantly either with the insulinaemic response or with cell surface receptor density. The findings suggest: (1) that a significant proportion of obese subjects are not hyperinsulinaemic and that their metabolic response to weight reduction may differ from that of those who are hyperinsulinaemic; (2) that reduced insulin binding to adipocytes in the obese reflects hyperinsulinaemia, when present, and not adiposity as such; (3) that the degree of insulinaemia is a major determinant of the cell surface receptor density.


Relationship of the activity of the bone isoenzyme of serum alkaline phosphatase to urinary hydroxyproline excretion in metabolic and neoplastic bone diseases. A significant correlation between the activity of the bone isoenzyme or serum alkaline phosphatase and the urinary hydroxyproline excretion in osteomalacia, osteoporosis, primary hyperparathyroidism with osteodystrophy, Paget's disease, secondary bone tumours, and in a control group was found (P less than 0.001). This close correlation was not observed between these variables in patients with active acromegaly. Diagnosis determined from these indices of formation and turnover of bone matrix agreed with that established by histological and histochemical examination of bone, by X-ray investigation of the skeleton, and by the radionuclear 85Sr test. The relationship between the activity of bone isoenzyme and urinary hydroxyproline excretion differed in metabolic bone diseases with a high bone turnover, in patients with osteoporosis and in patients with early osteoclastic bone metastases.


Studies on the turnover of endogenous cathodal trypsinogen in man. The concentration in serum of cathodal trypsinogen has been studied in certain clinical and experimental situations. The concentration correlated with pancreatic amylase activity. Low levels were found in patients with malabsorption due to exocrine pancreatic insufficiency. The concentration rose after endoscopic retrograde cholangiopancreatographic examinations (ERCP). After ERCP, however, no trypsin was detected complexed with protease inhibitors, as is generally found in acute pancreatitis. The trypsinogen concentration in serum also rose in renal failure indicating a renal elimination route for the endogenous trypsinogen.


Blood-brain barrier permeability during shortlasting intravascular hyperosmolality. Blood-brain barrier permeability for 24Na+ and 36Cl- ions and for 3HOH was studied during isoosmolality and during shortlasting intravascular hyperosmolality in twenty-five patients using the double indicatir single injection method. Hyperosmolality was induced by a rapid injection into the internal carotid artery of 8 ml of one of the following hypertonic solutions: 5% saline, 25% mannitol or a contrast medium of the metrizoat group (Isopaque-amine 280). The extractions of the small sodium and chloride ions remained unchanged and essentially zero indicating that 'opening' of the blood-brain barrier did not occur. The blood-brain barrier permeability to water remained unchanged during hyperosmolality, but the extraction of 3HOH increased when mannitol and especially when Isopaque-amine 280 was used as hypertonic agent corresponding to the lower water concentration in these solutions.


Quantitative changes of serum lipoprotein-X after cholestyramine administration in infants with cholestatic biliary tract and liver disease. Lipoprotein-X (LP-X) was determined before and after the administration of cholestyramine in fifty-five infants with persistent cholestatic jaundice to differentiate between intra- and extrahepatic disease. In twenty-seven infants with biliary atresia, serum LP-X prior to cholestyramine ranged from 0.87 to 11.42 g/l (mean: 3.43 g/l; the average concentration was significantly lower (P less than 0.001) in males. After cholestyramine, LP-X rose in twenty-three, remained the same in two, and decreased slightly in two infants. Serum LP-X was present in twenty of the twenty-eight infants with intrahepatic cholestasis prior to cholestyramine in concentrations from 0.84 to 14.19 g/l (mean: 3.13 g/l). After cholestyramine, LP-X decreased in all by an average of 78% (P less than 0.005). The other eight infants did not have LP-X before or after cholestyramine. This study shows that LP-X in the serum of infants with cholestatic jaundice indicates severe cholestasis, but is not itself diagnostic of biliary atresia. The differentiation of biliary atresia from other diseases is readily achieved, as the administration of cholestyramine for 2-3 weeks causes a marked decrease of serum LP-X in patients with patent extrahepatic bile ducts. The absence of serum LP-X excludes biliary atresia.


Increased excretion of two sialic acid-containing trisaccharides in the urine of patients with rheumatoid arthritis. The urinary excretion of sialic acid-containing trisaccharides in patients with active rheumatoid arthritis was studied. Sialyl-lactose and sialyl-N-acetyllactosamine were identified and their excretion patterns studied by thin layer and gas chromatography. The urinary output of sialyl-lactose was greater in patients with active rheumatoid arthritis (48.2 +/- 6.1 mg/24 h, SEM, n = 6) than in healthy subjects (19.8 +/- 3.7 mg/24 h, SEM, n = 5; P less than 0.01). The excretion of sialyl-N-acetyllactosamine was also higher in the rheumatoid group (18.5 +/- 2.1 mg/24 h, SEM, n = 6) than in the controls (11.1 +/- 1.2 mg/24 h, SEM, n = 5; P less than 0.05). The qualitative excretion patterns of the sialyl-oligosaccharide fraction were similar for the two groups as judged from the thin layer chromatograms. Correlating the results with the clinical state of the patients with rheumatoid arthritis suggests that the urinary level of the sialyl-oligosaccharides reflects the activity of the disease. A proposed mechanism for the increased excretion of sialic acid-containing trisaccharides in rheumatoid arthritis is presented.


Effect of portacaval shunt on biliary cholesterol and bile salt secretion in normal rats. In order to assess the role of bile secretion in the serum cholestrol lowering effect of a portacaval anastomosis in rats, bile flow rate, bile salt and cholesterol secretion in bile were determined. The study comprised five groups of animals: (1) rats with protacaval shunt cholesterol free food ad libitum. (2) sham-operated and pair-fed controls; (3) unoperated, pair-fed controls; (4) unoperated controls on cholesterol free diet ad libitum; and (5) unoperated controls on normal laboratory rat chow ad libitum. Bile flow rate showed no difference in any of the groups on cholesterol free diet. Bile salt secretion was significantly elevated in shunted animals (87.1 +/- 7.0 nmol/min) as compared with pair-fed sham-operated (64.7 +/- 20.0 nmol/min) and unoperated controls (56.0 +/- 22.3 nmol/min). Biliary cholesterol output tended to be higher in shunted animals but this was not statistically significant. The data indicate that the atrophied rat liver in shunted animals is able to maintain a normal bile flow and to increase bile salt secretion. Thus, the serum cholesterol lowering action of a portocaval anastomosis may be partly due to an increased cholesterol catabolism to bile salts.


Effect of continuously increasing concentrations of plasma ketone bodies on the uptake and oxidation of glucose by muscle in man. Muscle metabolism in man was studied by measuring the arterial and deepvenous concentrations of glucose, lactate, pyruvate, free fatty acids, beta-hydroxybutyrate and aceacetoacetate, and forearm blood flow. After the subjects had fasted overnight, their arterial free fatty acid and ketone levels rose continuously during a period of 90 min, leading to increased ketone body uptake by muscle. Hence, for each subject, a relation was obtained between arterial concentrations and arterial-deepvenous differences of beta-hydroxybutyrate and acetoacetate. As the ketone body utilization increased, the release of lactate rose as well. In spite of these alterations the uptake of glucose remained unchanged. These findings underline the current notion that accelerated ketone uptake reduces pyruvate oxidation but not glucose uptake by muscle.


Effect of muscle spasms on blood gases and acid-base status in tetanus patients. Blood gas tensions and acid-base status were determined in six patients with tetanus before and after the onset of severe muscle spasms. Hypoxaemia and metabolic acidosis present in all the patients, were worsened by muscle spasms. It is suggested that frequent measurements of blood gases should be carried out where such facilities are available.


Intrahypothalamic connections: an electron microscopic study in the rat. Terminal degeneration within the hypothalamus was studied by electron microsocpy 1 or 2 days (1) after carefully placed microlesions in the arcuate, anterior periventricular, ventromedial, premammilary and posterior hypothalamic nuclei and (2) after microlesions placed in the hypothalamus deafferented 3 weeks earlier. In the median eminence terminal degeneration was found after each of these lesions. Projections from the ventromedial nucleus reach the arcuate, suprachiasmatic, and anterior periventricular nuclei. Projections from the arcuate nucleus terminate in the medial preoptic, anterior periventricular, and ventromedial nuclei. After lesioning the premammillary nuclei degeneration was found in the supraoptic, arcuate, anterior hypothalamic and ventromedial nuclei.


I. Functional properties of neurons in lateral part of associative area 7 in awake monkeys. The lateral part of area 7, area 7b, of alert, behaving macaque monkeys was investigated using transdural microelectrode recording technique. Two hundred twenty-eight cells from five hemispheres of four monkeys were isolated and studied. The functional properties of 2% of the cells isolated remained unidentified. Functions of the identified cells were prominently related to the spatial control of arm movements. Of the cells 70% responded to somatosensory (40%) or visual (16%) or both somatosensory and visual (14%) stimulation. The receptive fields of these passively drivable cells were large, covering e.g., the arm or leg or chest or even the skin of the whole body. Most of the visually drivable cells responded to stimuli in both halves of the visual field. Of the cells responding to sensory stimulation 80% were activated by stimuli moving in a certain direction. Of the directionally selective cells 25% received information through more than one sensory channel. The complex stimulus-response relationships of these "convergence" cells revealed the existence of an integrative system which analyzes the direction of a stimulus moving in one sensory system using an other sensory system as a reference. Of all the cells isolated 28% discharged only during active movements of the arms (25%) or eyes (3%). Firing of these neurons was related to contraction of a functionally uniform group of muscles and not individual muscles. Some previous investigations of the parietal association cortex, conducted mainly in area 7a, have shown that most cells were active only when the monkey himself moves his eyes or arms. In our study on area 7b most cells responded to passive stimulation. The discrepancy between the results indicates functional differentiation within area 7.


II. Functional properties of cells in anterolateral part of area 7 associative face area of awake monkeys. The most anterior part of area 7 of awake, behaving macaque monkeys was investigated using single cell recording technique. Eighty-five cells from three hemispheres of two monkeys were isolated and studied. These cells showed more complex functional properties than the cells in the primary and secondary cortical fields. Of the cells 61% responded to somatosensory (26%) or visual (2%) or both somatosensory and visual (33%) stimulation; 39% of the cells were active only during the monkey's own movements. Most of the cells studied were active while the monkey was bringing an object to the mouth with its hand, when reaching for an object with lips, or while chewing. The neurons responded selectively to, e.g., palpation of the flexors of the arm, a visual stimulus approaching the face, passive movement of the monkey's hand towards the mouth, or they were active only when the monkey was reaching for an object with its lips or was mouthing it. The cellular activity in the anterolateral part of area 7 was prominently related to the stimulation or motor activity of the face (especially the mouth). In this respect, it differed from the more posterior part of area 7 adjacent to it. The results thus indicate that there is a separate and rather extensive mouth (or face) area in the parietal association cortex of the monkey.


Lability in the responses of cells in the auditory cortex of squirrel monkeys to species-specific vocalizations. The activity of 28 cells located mainly in the secondary auditory cortex (A II) of awake squirrel-monkeys, was extracellularly recorded for periods of up to 6 h. Seven different species-specific vocalizations, which were repeatedly presented to the monkey, were used as auditory stimuli. Twenty-six cells responded, at least once, to one or more vocalizations; 22 cells revealed some change in their response (pattern or strength) to at least one vocalization ("change in response"). Twenty-one cells exhibited a change in the number and/or type of vocalization to which they responded during the recording period ("change in selectivity"). At some time during the recording period all the responding cells exhibited a "change in response" and/or a "change in selectivity" ("change in responsiveness"). A "change in response" of a cell to a vocalization did not necessarily exclude a "change in selectivity", associated with the same vocalization, later in time and vice-versa. A "change in responsiveness" to one vocalization was not necessarily correlated with "changes in responsiveness" to other vocalizations.


The cingular vocalization pathway in the squirrel monkey. In 39 squirrel monkeys (Saimiri sciureus), the effects of various brain lesions on vocalizations elicited from the precallosal cingulate gyrus were tested. It was found that lesions abolishing the "cingular vocalization" completely can be traced from the stimulation site continuously down to the laryngeal motoneurons in the nucleus ambiguus. The pathway thus determined (Fig. 4) travels from the precallosal cingulate gyrus through the frontal white matter and enters the internal capsule from a dorsolateral position. The pathway then follows this structure in a medio-caudal direction down to the caudal diencephalon. Here, the effective lesions leave the corticospinal tract and ascend dorsally into the periaqueductal grey. The pathway follows this structure to its end where it sweeps lateral through the parabrachial area and then descends through the lateral pons and ventrolateral medulla to the nucleus ambiguus. In nine of the animals, in addition, the effects of bilateral anterior cingular lesions on vocalizations elicited in other brain areas were tested. It was found that the only vocalization-eliciting area which becomes ineffective after destruction of the anterior cingulate gyrus is the postero-medial orbital cortex.


Post-capillary venules in the lymphatic tissues of mice bearing experimental neoplasia. The lymph nodes, thymus and Peyer's patches of DBA/2 mice bearing an experimental tumor, mastocytoma, were assessed histologically with special reference to the structure of the post-capillary venules. For each of the lymphoid organ studied, the post-capillary venule score (PCV-S) was determined on the three grades (grades 1, 2 and 3) of the venules classified according to the height of the endothelial cells. The highest scores were obtained in the lymph nodes and Peyer's patches of the control animals. The scores in these lymphoid organs of the tumor-bearing mice were statistically highly significantly lower than in the control series. The lowest scores, however, were obtained in the nodes and patches of mice bearing mastocytoma after the previous treatment with anti-theta-globulin. The scores in the thymuses did not deviate from each other in the three series of mice studied. The findings of the present work support the concept that the structural state of the post-capillary venules in the lymph nodes and Peyer's patches is an important regulator of the T-lymphocyte recirculation in these organs. On the other hand, the venules of the thymus seem to be unrelated both structurally and functionally to the post-capillary venules of the nodes and Peyer's patches, and a new name of "junctional venules" has been proposed for these low endothelium walled venules of the thymus.


The use of cell free extracts derived from fungal protoplasts in the study of aflatoxin biosynthesis. A supernatant fraction derived from protoplasts of Aspergillus flavus was shown to be capable of converting both sterigmatocystin and versiconal hemiacetal acetate to aflatoxin B1. Versicolorin A was not converted under the same conditions.


Physical growth and brain development of captive-bred male and female squirrel monkeys, Saimiri sciureus. The physical growth and brain development of the captive bred male and female squirrel monkeys have shown that the male grows at a significantly faster rate with respect to these parameters in the postnatal life, starting from preweaning stage as compared to the female infant. During the prenatal life the male and the female fetuses grow at similar rates and show comparable brain development as indicated by its weight.


Metabolism of amphibian spermatozoa in relation to their motility. Bufo arenarum spermatozoa were able to sustain motility both under aerobic and anaerobic conditions. In aerobiosis, the oxygen consumption varies between 2.6 and 4.2 microliter O2/10(8) cells/h at 30 degree C. The synthesis of lactic acid by anaerobic spermatozoa demonstrated the existence of an active glycolytic pathway.


Spontaneous junctional currents in Drosophila muscle fibres: effects of temperature, membrane potential and ethanol. Fast, slow and fast multiquantal spontaneous junctional currents were recorded from glutamate sensitive muscle fibres of Drosophila larvae. Decrease of temperature and hyperpolarization prolonged the time course of fast currents. Ethanol (0.4 M) markedly shortened their duration, whereas several other drugs known to modify the time course of currents at cholinergic synapses were ineffective at this neuromuscular junction.


Hepatic UDP-glucuronyltransferase activity in acrylamide neuropathy. Acrylamide was found to be tissue selective in its toxicity. Doses which were clearly neurotoxic to rats were without effect on hepatic UDP-glucuronyltransferase, total hepatic protein or microsomal protein.


(N-Barbituryl)phenyl- and cyclohexylalkanoic acids and some of their derivatives as potential anti-inflammatory agents. A series of p-(N-barbituryl)phenyl- and cyclohexylalkanoic acids and their esters were synthesized. The preliminary pharmacological screening using the carrageenin, xylene and cotton tests revealed strong anti-inflammatory activity of several compounds in some of the tests performed, compound (Ij) being active in all the tests.


Dynamic evaluation of prolactin secretion in patients with oligospermia: effects of treatment with metergoline. Serum prolactin levels were measured in 50 patients with oligospermia and in 20 control subjects under fasting conditions and following the administration of levodopa, pyridoxine, metoclopramide, and synthetic thyrotropin-releasing hormone. Four patients (8%) under fasting conditions had prolactin levels slightly above the normal range. However, no significant differences in prolactin behavior were detected between patients with hyperprolactinemia, patients with normal prolactin levels, and control subjects. The four patients with hyperprolactinemia were treated with metergoline, an ergoline derivative. Metergoline administration promptly reduced the prolactin levels. Spermatogenesis was restored in three patients after 4 to 5 months of treatment.


[Piecewise Fourier analysis of images and the role of the occipital, temporal and parietal cortex in visual perception]. On the basis of previously published data, a neuronal scheme of organization of the visual perception in the cerebral cortex, is suggested. An idea of neuronal module is introduced: a cylinder of cortical neurons whose receptive fields are directed towards the same area of the visual field and respond to different spatial frequencies and orientations. A system of overlapping modules is able of piece--wise Fourier--description of portions of the image. The modules of the Clare--Bishop area are composed of receptive fields of different size. Owing to that each neuron of the module projects inhibitory influence upon other neurons, the module acts as a filter picking out the texture. Therefore, the modules of the Clare--Bishop area single out and supply Fourier--description of subimages, the latters being characterized by the same local spectrum within their own limits. The lower portiön of temporal cortex performs a rough identification of subimages and images with the aid of systems of learning neurons. The parietal cortex conforms the description from temporal cortex to the complete description in the modules of occipital cortex, thus performing the transition from an image to a concrete picture.


Occurrence and significance of antibody to liver-specific membrane lipoprotein by double-antibody immunoprecipitation method in sera of patients with acute and chronic liver diseases. A double-antibody immunoprecipitation method was developed for detecting antibody to liver-specific membrane lipoprotein (anti-LSP) in sera of patients with various liver diseases and primary nonhepatic autoimmune diseases. Liver-specific membrane lipoprotein prepared from normal rat livers was labeled with 125I (chloramine-T) and monospecific antibody raised in rabbits. Cross-reactivity and absorption studies demonstrated that the assay used was highly specific. The frequency and titer of anti-LSP were similar for HBsAg-positive and -negative patients with both acute and chronic liver diseases. Patients with chronic active hepatitis had the highest frequenzy (25 of 44 cases, 57%) when compared with those with chronic persistent hepatitis (5 of 23 cases, 22%) and nonalcoholic cirrhosis (8 of 21 cases, 38%). Of the anti-LSP positive cases, the mean titer in patients with chronic active hepatitis tended to be the highest. In patients recovered from acute viral hepatitis, anti-LSP was transiently positive (7 of 20 cases, 35%) in the acute phase. In those who progressed to chronic hepatitis, a late rise as well as an early rise occurred in 6 of 10 patients before the diagnosis was made. Two of 6 patients with primary biliary cirrhosis had anti-LSP, but none of 41 patients with other nonviral liver diseases and none of 60 patients with primary nonhepatic autoimmune diseases. These data indicate that an autoimmune reaction directed against LSP can be initiated during the acute phase of viral hepatitis and it may persist in chronic hepatitis in both HBsAg-positive and -negative cases.


Regulation of newly evolved enzymes. IV. Directed evolution of the Ebg repressor. In Escherichia coli, the wild-type repressor of ebg (evolved beta-galactosidase) enzyme synthesis, specified by the ebgR+ gene, responds very weakly to lactulose (fructose-beta-D-galactopyranoside). Selection for a functional repressor that responds strongly to lactulose as an inducer reveals the existence of ebgR+L mutants, which occur spontaneously at a frequency of about 2 X 10(-10) . EBGR+L mutants are pleiotropic in that they specify ebg repressor with a greatly increased response to lactulose, lactose, galactose-arabinoside and methyl-galactoside as inducers. Selection of ebgR+L mutants is discussed within the framework of directed evolution of a regulatory function.


Two closely linked mutations in Drosophila melanogaster that are lethal to opposite sexes and interact with daughterless. A new spontaneous mutation named Sex-lethal, Male-specific No. 1 (SxlM1) is described that is lethal to males, even in the presence of an Sxl+ duplication. Females homozygous for SxlM1 are fully viable. This dominant, male-specific lethal mutation is on the X chromosome approximately 0.007 map units to the right of a previously isolated female-specific mutation, Female-lethal, here renamed Sex-lethal, Female-specific No. 1 (SxlF1). SxlM1 and SxlF1 are opposite in nearly every repect, particularly with regard to their interaction with maternal effect of the autosomal mutation, daughterless (da). Females that are homozygous for da produce defective eggs that cannot support female (XX) development. A single dose of SxlM1 enables daughters to survive this da female-specific lethal maternal effect. A duplication of the Sxl locus weakly mimics this action of SxlM1. In contrast, SxlF1 and a deficiency for Sxl, strongly enhance the female-lethal effects of da. The actions of SxlM1 and SxlF1 are explained by a model in which expression of the Sxl locus is essential for females, lethal for males, and under the control of a product of the da locus. It is suggested that SxlM1 is a constitutive mutation at the Sxl locus.


The effect of recombination-defective meiotic mutants on fourth-chromosome crossing over in Drosophila melanogaster. Crossing over was measured on the normally achiasmate fourth chromosome in females homozygous for one of our different recombination-defective meiotic mutants. Under the influence of those meiotic mutants that affect the major chromosomes by altering the spatial distribution of exchanges, meiotic fourth-chromosome recombinants were recovered irrespective of whether or not the meiotic mutant decreases crossing over on the other chromosomes. No crossing over, on the other hand, was detected on chromosome 4 in either wild type or in the presence of a meiotic mutant that decreases the frequency, but does not affect the spatial distribution, of exchange on the major chromosomes. It is concluded from these observations that (a) in wild type there are regional constraints on exchange that can be attenuated or eliminated by the defects caused by recombination-defective meiotic mutants; [b] these very constraints account for the absence of recombination on chromosome 4 in wild type; and [c] despite being normally achiasmate, chromosome 4 responds to recombination-defective meiotic mutants in the same way as do the other chromosomes.


[Operon of riboflavin biosynthesis in Bacillus subtilis. XV. A study of mutants related to the initial stages of biosynthesis. The origin of the ribityl chain of the riboflavin molecule]. The incorporation of 14C-labelled guanosine and xanthosine into riboflavin was studied. It is concluded that the ribose mojety of guanosine is converted to the ribityl side chain of riboflavin. Thus the immediate precursor of riboflavin biosynthesis is a guanosine compound. Two classes of the riboflavin-dependent mutants of Bacillus subtilis were studied. They are closely linked to the lysine markers and probably correspond to the initial steps of riboflavin biosynthesis pathway.


[Ontogenetic switchover in Bacillus subtilis. II. The dynamics of the stationary phase processes during growth under conditions of catabolite repression]. To reveal the pecularities of the growth under the conditions of catabolite repression (medium 2) of Bacillus subtilis and the mutants obtained, the investigations of dynamics of the following processes were carried out: alteration of the pH of the culture exhaustion of glucose in the medium, appearance of the activity of both aconitase in the cells and extracellular metal- and serine proteases in the supernatant, and the appearance of the thermoresistant spores. The following features were observed during the growth under the conditions of catabolite repression: 1. Bacillus subtilis WB 746 and cgs mutants: the death of the main part of the culture after the Iogarithmic phase of growth (LPG), the presence of the secondary LPG of the survived cells which have the increasing activity of aconitase, the appearance and sharp increase in the extracellular serine protease activity 6 hours before thermoresistant spore formation. In the case of cgs mutants the activity of metal proteases appears and increases during the secondary LPG; 2. In the culture of cgl mutants the pH is lowered to 5.1 at the end of the LPG and after the glucose exhaustion the death of almost all the culture follows; 3. cgr mutants: a comparatively high activity of aconitase in the cells is found by the time of the early LPG, and at the end of the LPG the activity of both metal- and serine proteases appear in the supernatant of the culture and the secondary induction of the serine protease activity 6 hours before thermoresistant spore formation is observed. The serine protease activity found in the supernatant before and after the secondary induction of the enzyme belongs to the identical protein. During the stationary phase of the growth of cgr mutants, the high rate of 3H-uridine incorporation into the RNA molecules which have the electrophoretic mobility of mRNA was observed. The sporulation of Bac. subtilis strains under investigation, except cgl mutants, occurs when the culture has reached the definite state: the alkaline pH, the presence of the aconitase activity in the cells and the induced activity of serine protease.


A new sequence-specific endonuclease from Streptococcus faecalis subsp. zymogenes. A new sequence-specific endonuclease, SfaI, has been partially purified from Streptococcus faecalis subsp. zymogenes. SfaI recognizes the tetranucleotide sequence 5'G-G-C-C 3' 3' C-C-G-G 5' and cleaves it at the sites indicated by the arrows.


Controlled trial of disodium cromoglycate in chronic persistent ulcerative colitis. Oral disodium cromoglycate (200 mg qds) has been tested in 26 patients with ulcerative colitis that was resistant to medical treatment. In a double-blind crossover trial disodium cromoglycate and placebo were added to conventional treatment in random order, each for four weeks. There was no significant difference in therapeutic effect between disodium cromoglycate and placebo.


Gamma heavy chain disease simulating alpha chain disease. A young Turkish girl presented with all the clinicopathological features of a digestive form of alpha chain disease. A gamma heavy chain disease protein, however, was found in her serum and also in the cells invading the intestinal mucosa and mesenteric lymph nodes.


Contribution to the serological diagnosis of sarcocystosis. The importance of the IFA-test (Indirect Fluorescent Antibody Test) in the diagnosis of sarcocystosis in the intermediate host has been confirmed in the present paper. We assessed the time of the first appearance of serum antibodies in the intermediate host in mice inoculated experimentally with the species Sarcocystis dispersa. By means of the IFA-test, the first antibodies were found on day 20 p.i. Cross-reactions among antisera of S. dispersa and a heterologous antigen of S. cernae disclosed that the reaction was not species-specific, but genus-specific. In addition, we confirmed serologically that the antigenic structure of the genus Frenkelia was identical to that of the genus Sarcocystis, because the results of cross reactions obtained with the IFA-test were identical.


[Bronchodilating action of (--)-1-(3, 4, 5-trimethoxybenzyl)-7-hydroxy-1, 2, 3, 4-tetrahydroisoquinoloine HCl (l-MTI) (author's transl)]. Bronchodilating action of l-MTI was investigated by intravenous administration to anesthetized animals and the results were compared to those seen with isoproterenol. Bronchoconstriction was induced by histamine or serotonin. Under pentobarbital anesthesia, the bronchodilating activity of l-MTI was approx. 1/20 that of isoproterenol in cats and dogs, but less than 1/100 that of isoproterenol in monkeys and guinea pigs. The results indicate that the bronchodilating activity of l-MTI is variable and species dependent. It was demonstrated, however, that in guinea pigs, l-MTI did not produce any bronchodilating activity, when the animals were anesthetized with urethane. The absence of the bronchodilating response in animals under this anesthesia was also seen in vagotomized, adrenalectomized and reserpinized, norepinephrine-infused guinea pigs. On the contrary, l-MTI exhibited the bronchodilating action in reserpinized, pithed or hexamethonium-treated guinea pigs anesthetized with urethane. On th other hand, isoproterenol produced a bronchodilating action in all preparations mentioned above. It was also evident that l-MTI suppressed the bronchodilating action of isoproterenol in pentobarbital anesthetized guinea pigs. In addition, l-MTI increased the heart rate of guinea pigs with a low beating rate, but decreased it when the control rate was high. These results suggest that at least in guinea pigs, l-MTI acts as a partial agonist and that the effect of l-MTI on the bronchial system depends on the sympathetic tone and/or sympathetic bronchodilator reflex, as proposed by McCULLOCH ET AL. (9).


Effect of inhibitors on acid production by baker's yeast. Glucose-induced acid extrusion, respiration and anaerobic fermentation in baker's yeast was studied with the aid of sixteen inhibitors. Uranyl(2+) nitrate affected the acid extrusion more anaerobically than aerobically; the complexing of Mg2+ and Ca2+ by EDTA at the membrane had no effect. Inhibitors of glycolysis (iodoacetamide, N-ethylmaleimide, fluoride) suppressed acid production markedly, and so did the phosphorylation-blocking arsenate. Fluoroacetate, inhibiting the citric-acid cycle, had no effect. Inhibition by uncouplers depended on their pKa values: 2,4,6-trinitrophenol (pKa 0.4) less than 2,4-dinitrophenol (4.1) less than azide (4.7) less than 3-chlorophenylhydrazonomalononitrile (6.0). Inhibition by trinitrophenol was only slightly increased by its acetylation. Cyanide and nonpermeant oligomycin showed practically no effect; inhibition by dicyclohexylcarbodiimide was delayed but potent. The concentration profiles of inhibition of acid production differed from those of respiration and fermentation. Thus, though the acid production is a metabolically dependent process, it does not reflect the intensity of metabolism, except partly in the first half of glycolysis.


Eurotium (Aspergillus) repens metabolites and their biological activity. Eurotium repens mycelium cultivated under static conditions was used to isolate and identify metabolities--echinulin, physcion, erythroglaucin, flavoglaucin and asperentin; the filtrate of the culture yielded asperentin 8-methylether. The broadest biological activity spectrum was displayed by asperentin which had antibacterial and antifungal effects and, at a concentration of 86 microgram/ml, caused 50% mor7 tality in Artemia saline larvae. The highest cytotoxicity towards HeLa cells was found in physcion which caused 50% growth inhibition at a concentration of 0.1 microgram/ml.


Inhibition of photo-induced Trichoderma viride conidiation by inhibitors of RNA synthesis. The photo-induced conidiation of Trichoderma viride is suppressed by ethidium bromide, acriflavin, lomofungin and 8-quinolinol at concentrations which do not inhibit the colony growth of this deuteromycete.


[Acute polyneuropathy due to phosphate deficiency during parenteral feeding]. A malnourished M. Crohn-patient developed severe symptoms similar to those of an acute polyneuropathy associated with a period of marked decrease of serum phosphate concentration during intravenous high-caloric infusions. After the serum phosphate concentration was normalized recovery from all neurologic symptoms followed within a few days. Based on this observation the possible consequences of severe hypophosphatemia are discussed.


Thyroxine inactivation by starvation in cultured hepatocarcinoma cells, Formation of reverse triiodothyronine. The effect of starvation on thyroid hormone metabolism was studied in monkey hepatocarcinoma monolayer cultures. Nonphenolic ring monodeiodination of thyroxine, 3, 5, 3'-triiodothyronine and 3, 3'-diiodothyronine was accelerated. Since phenolic ring deiodination of 3, 3',5'-triiodothyronine (reverse T3) was unaffected, this metabolite accumulated in the medium during thyroxine metabolism. This suggests that increased serum reverse T3 in malnourished humans may be caused by enhanced deiodination of thyroxine rather than decreased rT3 catabolism.


Canada's NHI: a physician's evaluation.

Canada's NHI: a physician's evaluation. A physician in Manitoba reports that a national health care system has worked well in Canada, particularly for poor people and for governments concerned about the equitable and universal delivery of health care, as well as about rising health costs. Physicians, however, find the greatest problems are their own low salary and their lack of impact on government policy.


Variability of bacterial gene-directed enzyme production in human genetically deficient cells. Human beta-galactosidase-deficient skin fibroblasts from a patient with generalized gangliosidosis (GMI-gangliosidosis type I) were treated with phage lambda plac DNA, coding for Escherichia coli beta-galactosidase (beta-D-galactoside galactohydrolase, EC.3.2.1.23). New beta-galactosidase activity detected in cell extracts of phage DNA-treated GMI-gangliosidosis fibroblasts continued to vary considerably from one experiment to another. It behaved like the E. coli z-gene product upon immunochemical and physicochemical investigation. In some experiments the antigenic behavior of resultant beta-galactoside activity in lambda plac DNA-treated cells resembled that of mutant E. coli beta-galactosidase. Among the factors and variables that may be responsible for the variation in the results obtained here and elsewhere, low physical binding between prokaryotic mRNA sequences and fibroblast ribosomal RNA could play a part connected with effective translation. This hypothesis is discussed under the aspect of a comparison of the ribosomal binding site of lac z mRNA with the 3'-terminus of the eukaryotic 18s ribosomal RNA, which shows limited possibilities for base-pairing interactions. More extensive possibilities for forming Watson-Crick base pairs between their initiation site and the eukaryotic ribosomal binding site exist for other prokaryotic messengers, such as those of Q beta-replicase, f 1-coat protein, or UDPG-4-epimerase.


Anti-kappa/lambda immunobead for detection of human B lymphocytes. Several methods were compared for quantitating the B lymphocytes in human peripheral blood. RBC rosette assays including sheep and bovine red blood cells, and immunobead rosette assays including beads coated with anti-kappalambda and trivalent anti-GAM antibodies respectively were investigated. The anti-kappalambda bead assay proved to be simpler, faster, higher in reproducibility, and appeared to be more accurate than any of the other assays.


[False-negative antiglobulin-(Coombs-) test caused by anticomplement bovine albumin batches in the transfusion preparation]. Different brands of bovine albumins were tested as reagent in the second step of the so called 3-stage-screening test for irregular blood group antibodies. In 3 of 9 brands of bovine albumin there was anticomplementary activity which was the cause of diminished or negative reactions in the third stage of the screening test (antiglobulin stage). The disadvantageous brands of bovine albumin contained Na2-EDTA which revealed to be responsible for the anti-complementary activity.


[The level of the plasma proteins transferrin, retinol-binding protein and prealbumin in the postoperative parenteral feeding by administration of varying amounts of amino acids]. 38 adult patients of a surgical intensive care ward received differently large amounts of amino acids during the course of postoperative parenteral nutrition. The dose of amino acids was adapted to the preoperative nutritional condition of the patient. Changes in the serum levels of retinol-binding protein, prealbumin and transferrin were registered as well as changes in total protein and nitrogen balance up to a maximum of 10 postoperative days. Depending upon the amount of administered amino acids, the values for transferrin and nitrogen balance had changed more significantly than those of serum concentrations of retinol-binding protein and of prealbumin.


[Postoperative long-term feeding with maltose supplementation]. 2 litres of a 40% carbohydrate combination solution consisting of glucose, fructose, xylitol and maltose in the ratio 1:1:1:1 were infused together with 1 litre of 8% amino acid solution (aminomel L 8 oKH salvia) over a period of 24 hours for 4 days to 16 patients who had undergone gastric resection by the Billroth II procedure. Since maltose is partly excreted as glucose, maltose and glucose were jointly balanced. The joint conversion was, on average, 75, the fructose conversion 97 and the xylitol conversion 94%. The mean conversion of the 4 carbohydrates was about 85% corresponding to 2800 kcal (11,720 joule) daily. The nitrogen balance showed initally a slight catabolism. The serum insulin level rose significantly and simultaneously there was inhibition of lipolysis. The metabolic state and tolerance parameters remained in the normal range. The only side-effects noted were an increasing activity of the alkaline phosphatase and a significant elevation of the gamma-GT on the 4th day. The administration of relatively large quantities of maltose and glucose in combination with fructose and xylitol does not appear to produce any notable advantages.


Microscopic characterization of rabbit lung damage produced by Pseudomonas aeruginosa proteases. The intratracheal administration of highly purified Pseudomonas aeruginosa proteases (ca. 10 to 100 microgram) elicited extensive, grossly observable rabbit lung damage by 3 h postinjection. Light and electron microscopic characterization of the lesions revealed: (i) progressive injury and necrosis of type I epithelial cells and capillary  endothelial cells from 3 h to 1 day postinjection, and progressively increasing accumulations of erythrocytes, plasma proteins, fibrin, and released type II epithelial cell lamellar bodies in alveolar lumina during that time period; (ii) progressively increasing accumulations of macrophages, but not of polymorphonuclear leukocytes, in alveolar lumina from 3 h to 6 days postinjection; (iii)  progressive hyperplasia of type II epithelial cells from 12 h to 4 days postinjection; (iv) progressive infiltration of alveolar septa by mononuclear inflammatory cells (interstitial pneumonitis) from 2 to 6 days postinjection; (v) no loss of alveolar septal connective tissue and no damage to pulmonary arterioles and venules; and (vi) almost normal alveolar structure by ca. 8 days postinjection. The study revealed that the intra-alveolar hemorrhage, the injury and necrosis of alveolar septal cells, and the infiltration by mononuclear cells that have been reported to occur during human pseudomonas pneumonia can also be elicited by the experimental administration of pseudomonas proteases. Thus, the results support the idea that in vivo production and activity of P. aeruginosa proteases is important, at least in part, in eliciting the lung damage characteristic of pseudomonas pneumonia.


Anticomplementary activity of tuberculin: relationship to platelet aggregation and lytic response. Experiments were performed to examine the interaction of tuberculin with platelets and complement. Hemolytic complement titrations show that tuberculin consumes complement in human, rabbit, and guinea pig serum. Evidence in support of classical pathway activation was provided by observation of C1 consumption and failure to detect significant conversion of alternative pathway factor B to B by immunoelectrophoresis. Platelets in plasma from guinea pigs deficient in the fourth component of complement were not affected by tuberculin. However, studies on platelet aggregation in plasma chelated with ethyleneglycolbis(beta-aminoethyl ether)-N,N-tetraacetic acid indicated that tuberculin may initiate sluggish activation of the alternative pathway. That the reaction between tuberculin and platelets is a lytic one was evidenced by observing the release of the cytoplasmic enzyme lactic dehydrogenase and efflux of rubidium-86. Studies with C6-deficient rabbits indicated that platelet release of exogenously supplied tritiated serotonin is caused by platelet lysis.


Microbial interference and colonization of the murine gastrointestinal tract by Listeria monocytogenes. Two strains of Listeria monocytogenes, one that formed smooth colonies on agar surfaces and a varient of it that formed rough colonies, colonized the gastrointestinal tracts of germfree mice. Within 24 h after mice were inoculated orally with about 100 bacteria, the population levels per gram (wet weight) of tissue of both strains were 10(5) to 10(7) in the stomach and ileum and 10(8) to 10(9) in the cecum and colon, respectively. As detected in Gram-stained histological sections, in such gnotobiotes, the bacteria colonized the lumen in all areas of the tract and much of the mucus layer on the epithelial surface in the proximal colon. The strain that formed smooth colonies did not colonize the tracts of specific-pathogen-free mice, but did colonize, to the same levels as in germfree mice, the stomachs and bowels of ex-germfree mice previously associated with two members of the indigenous flora (Bacteroides and Clostridium). In the latter animals, however, the listeria did not form layers on the colonic epithelium as efficiently as they did in monoassociated gnotobiotes.


Enhancement of resistance to murine osteogenic sarcoma in vivo by an extract of Brucella abortus (Bru-Pel): association with activation of reticuloendothelial system macrophages. The administration of an aqueous-ether extracted residue of Brucella abortus (Bru-Pel) inhibits development of transplanted osteogenic sarcomas in mice as evidenced by a decrease in mortality. At least one mechanism through which Bru-Pel modulates host resistance is activation of macrophages of the reticuloendothelial system. Peritoneal macrophages harvested from mice receiving Bru-Pel were cytotoxic for osteogenic sarcoma cells in vitro, limited the replication of vaccinia virus in cell cultures, and demonstrated enhanced emittance of chemiluminescence during phagocytosis of zymosan particles of Candida albicans. The concept of reticuloendothelial system activation was further supported by the evidence that administration of Bru-Pel enhanced resistance of mice to challenge with a lethal inoculum of Listeria monocytogenes. These observation support the hypothesis that Bru-Pel shares a number of characteristics with recognized immunomodulating agents and that one mechanism by which it modulates host resistance to tumors, to virus infections, and to challenge with L. monocytogenes is through activation of macrophages.


Binding of antigen-enzyme complex by antigen-binding cells as an approach for immunogdiagnosis. An immunoenzyme method capable of detecting specific antigen-binding cells is developed. The method is based on the chemical coupling of the antigen to beta-galactosidase and binding of the conjugate to the corresponding antigen-binding cells. Human hydatid fluid was used as an antigen model, and cells derived from rabbits immunized with human hydatid fluid showed specific binding to human hydatid fluid-beta-galactosidase conjugate. The method is specific and reproducible and could be used as an immunodiagnostic test for various diseases in which little or no antibody is produced.


Antitumor activity of Listeria monocytogenes on a guinea pig fibrosarcoma. Listeria monocytogenes-mediated tumor inhibition was studied in strain 13 guinea pigs by using a methylcholanthrene-induced fibrosarcoma (MCA-1). Mixtures of Listeria and tumor cells in ratios of 1:100, 1:200, or 1:400 (Listeria:MCA-1 cells) led to significant suppression of tumor growth. Intralesional injection of tumors on day 6 posttransplantation led to the regression of a highly significant number of tumors. Animals receiving injections of Listeria, either in a mixture with tumor cells or intralesionally, displayed enhanced skin test reactivity to a tumor extract. Tumor regressors were resistant for at least 2 to 3 months after the initial transplant to rechallenge with MCA-1 cells. Thus, with this particular tumor-host system, Listeria was successfully employed as an antitumor agent with no visibly detrimental side effects to the host.


Growth of Rickettsia typhi in irradiated L cells enhanced by lysosomal stabilization. The growth of some obligate intracellular parasites is contingent upon avoidance of lysosomal activation during growth in their host cells. This is accomplished by the various parasites by different mechanisms and with different degrees of efficiency. The possibility was tested that the lysosomal stabilizer cortisone acetate might protect and thus enhance the growth of Rickettsia typhi in mouse L cells irradiated 6 days earlier. Beginning 2 days before infection of the L cells with a multiplicity of 10 rickettsiae, 20 microgram of cortisone per ml was added in medium 199 containing 5% fetal calf serum. This concentration of cortisone was below the cytotoxic level, as determined by viability staining, but was sufficient to significantly alter the ratios of cellular and released acid phosphatase and beta-glucuronidase in uninfected and infected cells, as shown by spectrophotometric analysis. Rickettsial replication, measured by hemolytic activity at 96 h and confirmed by microscopic observations at earlier stages of infection, was increased by the cortisone. Cortisone concentrations of 10 or 40 microgram/ml were less effective, and cortisone was ineffective when the rickettsial multiplicity per L cell was 2 or lower. These results indicate that amounts of cortisone that increase lysosomal stabilization in L cells favor rickettsial multiplication when the multiplicity of infection is relatively high.


Phage-related surface modifications of Pseudomonas aeruginosa: effects on the biological activity of viable cells. Lysogenic [EI(8)3] and phage 8-resistant mutant (EI/8S17) strains of Pseudomonas aeruginosa EI were isolated. Besides lacking the capacity to adsorb phage 8, strains EI(8)3 and EI/8s17 did not contain surface substrate for the depolymerase that is produced de novo when phage 8 infects wild-type strain. EI. The glycolipoprotein (GLP) in the wild type contains phage 8 receptors and surface substrate for the depolymerase, as well as possesses characteristics of a virulence factor; therefore, the chemical and biological characteristics of the derived strains were investigated. The neutral-sugar, amino sugar, and protein content of the GLPs from the derived strains differed quantitatively from that of the wild type. In spite of some cross-reactivity, the GLPs from all strains were antigenically distinct in the indirect hemagglutination inhibition test. In mice, the toxicity of the GLP from strain EI(8)3 equaled that of the wild type, but the  GLP of strain EI/8s17 was threefold less toxic.  Significantly fewer viable EI(8)3 cells were required for the mouse 50% lethal dose than for the cells of either the wild type or the phage-resistant mutant.


Antibody responses in the cerebrospinal fulid of cynomolgus monkeys after intracerebral inoculation with paramyxoviruses. Cynomolgus monkeys with or without measles antibody were intracerebrally inoculated with measles or canine distemper viruses, and antibody responses in the cerebrospinal fluid (CSF) were investigated. In measles antibody-free monkeys, natural infection with wild measles virus or intracerebral inoculations with two attenuated measles vaccines evoked primary antibody responses to measles virus in the sera but not in the CSF. In measles-immune monkeys, intracerebral inoculation with the TYCSA strain of measles virus produced a significantly high titer of measles antibody in the CSF with a minimal rise in the serum antibody and resulted in a significant decrease in serum/CSF antibody ratios. Intracerebral inoculation of a neurotropic canine distemper virus, the Onderstepoort strain, into measles-immune monkeys caused production of both measles and distemper antibodies in the CSF. Inoculation of measles-immune monkeys intravenously with measles virus or intracerebrally with rubella virus, which has no antigenic relation to measles virus, failed to evoke a measles antibody response in the CSF. These results indicated that local production of measles antibody in the CSF was caused by a stimulus within the central nervous system of measles virus antigen or canine distemper virus antigen that partially cross-reacted with measles virus antigen as a secondary antibody response.


Macrophage activation during experimental murine brucellosis: a basis for chronic infection. Evidence is presented that the chronicity of infection in CBA mice after injection of Brucella abortus 19 is related to a number of factors: (i) the relative resistance of B. abortus to macrophage killing, which allowed some bacteria to survive the peak of macrophage activity occurring at 14 days; (ii) the decline in macrophage activity thereafter (this decline was related in part to the presence of fewer bacteria to stimulate the bactericidal activity and also to specific, active suppressor mechanisms not identified in this study); and (iii) the insensitivity of the persistent Brucella organisms to activated macrophages. This was not due to a selection of genetically resistant bacteria, but possibly to their inaccessibility, either within "incompetent" macrophages or outside macrophages altogether.


Defective cellular immunity to gram-negative bacteria in cystic fibrosis patients. In vitro lymphocyte responses to Pseudomonas aeruginosa have been found to be impaired in cystic fibrosis patients with advanced clinical disease. The responses to Klebsiella pneumoniae, Serratia marcescens, and Proteus mirabilis were studied in a similar group of cystic fibrosis patients and normal individuals. Cystic fibrosis patients found to be unresponsive to pseudomonas were also unresponsive to klebsiella, serratia, and proteus. Responsiveness to Staphylococcus aureus was not impaired in cystic fibrosis patients. We postulate that in vitro lymphocyte responses to several gram-negative bacteria require the function of a lymphocyte subpopulation which may be impaired in some cystic fibrosis patients.


Immunofluorescent studies of the local immune response in the mammary glands of rats. Two irritants, phosphate-buffered saline and alcohol, and antigens including killed Brucella abortus, live B. abortus, Staphylococcus aureus, and rat parvovirus were separately infused into rat mammary glands during pregnancy, and by using immunofluorescent techniques, the numbers of immunoglobulin-containing cells in glands during lactation and involution were determined. The study provided basic information on the local immune response of the mammary gland to antigens of various types. In all experiments, the number of immunoglobulin M (IgM) cells present was small and no trends were apparent. IgA cells were always more prevalent than IgG cells. Fewer IgA cells were in the glands of rats infused with phosphate-buffered saline and alcohol than in normal rats. IgA cell prevalence was greatest in response to infusion of live B. abortus. Responses to live S. aureus and parvovirus were less pronounced, and infusion of killed B. abortus did not induce an elevation in IgA cell prevalence. IgG cell prevalence was greatest in response to infusion of live B. abortus or S. aureus and was decreasingly less pronounced in response to killed B. abortus and rat parvovirus. With the exception of parvovirus infusion, in regard to IgA cells, all glands locally infused with antigen had elevated IgA and IgG cell numbers when compared with noninfused glands in the same animal.


Passive immunization against Pseudomonas with a ribosomal vaccine-induced immune serum and immunoglobulin fractions. Passive protection of mice against Pseudomonas aeruginosa using specific antisera and immunoglobulin fractions induced by immunizing rabbits with a ribosomal vaccine is reported. The results demonstrated that protection by the ribosomal vaccine against challenge with live organisms can be serum mediated. Previous work has shown that the vaccine can be separated into two components on the basis of molecular weight and that both higher (peak A)- and lower (peak B)-molecular-weight fractions were capable of inducing active immunity in mice. The present report indicates that both fractions are also capable of eliciting the production of mouse-protective antibody in rabbits. Agar gel diffusion with antisera to peaks A and B or unfractionated vaccine indicated a common antigenic component among them in addition to an extra antigen in unfractionated vaccine not present in peak B. Passive hemagglutination with antisera to peaks A and B demonstrated high-titer agglutinating antibody only with antiserum to peak A when a method of erythrocyte sensitization for lipopolysaccharide antigens was used. Also, passive hemagglutination was greatly inhibited by small amounts of lipopolysaccharide prepared from the same organism from which the vaccine was made. Both antisera to peaks A and B fixed complement with either A or B antigens. Antisera to peaks A and B, when reacted with peak B antigen, had about the same complement fixation titer (as determined by a quantitative complement fixation test). However, when peak A antigen was used, antiserum to peak A had about twice the complement fixation titer that antiserum to peak B had. These results are consistent with previous observations which suggest that the ribosomal vaccine contains lipopolysaccharide in addition to an unidentified immunogenic principle associated with ribosomes. Furthermore, this immunogen was present in both peaks A and B, but detectable amounts of lipopolysaccharide were present only in peak A. The relative importance of the immunoglobulin G (IgG) and IgM classes of antibodies was also compared. The results indicated that both IgG and IgM isolated from immune rabbit serum are protective in mice. Only IgG precipitated with the vaccine in agar gel diffusion, but both IgG and IgM were active in passive hemagglutination and in complement fixation. The passive hemagglutination titer of the IgM was higher than that of the IgG, but the complement fixation titer of the IgG was higher than that of the IgM. The mouse-protective capability of the IgG and IgM was about the same.


Group and type antigens of Capnocytophaga. Group-specific and type-specific antigens have been identified and purified from sonic extracts of Capnocytophaga. The group-specific antigen, which was purified by affinity chromatography, was found to be identical to an antigen present in all 26 strains tested. The antigen is sensitive to trypsin, sodium dodecyl sulfate and heat labile, and composed predominantly (55%) of protein. The type-specific antigen, which was obtained by preparative immunoelectrophoresis, was found to be present in only 3 of the 26 strains tested. This antigen was resistant to heat, trypsin, and sodium dodecyl sulfate and was primarily composed of carbohydrate (47% phenol-sulfuric acid-positive material, 8% amino sugar). Agglutination and fluorescent antibody data suggest that both the group- and type-specific antigens reside on the cell surface.


A new assay for the measurement of mediator release from rat peritoneal mast cells. In this report, we describe a new in vitro experimental system for monitoring the release, and inhibition of release, of [3H]-serotonin from rat peritoneal mast cells, which can be used as a novel screen for evaluation of compounds effective in immediate hypersensitivity reactions. This assay is an improvement on existing assays since the cells are not required to be exposed to hormones at any time during the procedure. In this assay, 1 microgram/ml of compound 48/80 consistently produced a 6- to 8-fold-increase in release of serotonin with a low background release of 4-5%. Disodium chromoglycate (DSCG), a standard inhibitor of histamine release, effectively inhibited compound-48/80-stimulated serotonin release with an average I50 of 2.1 X 10(-4) M. Several other potential antiasthmatic agents were far more potent than DSCG.


Effects of the obese (ob/ob) genotype on spleen cell immune function. Spleen cells from mice homozygous for the obese (ob) mutation killed DBA/2 mastocytoma target cells less well than spleen cells from lean littermates or unrelated age-and sex-matched controls of the same strain. Killing was impaired only when the attacker cells were primed in vivo, not following in vitro priming. Hence the effect of the ob/ob genotype is not to produce an irreversible functional change in the lymphocyte, but rather to produce an environment in which lymphocytes are less able to react to priming antigen. Not only were the spleen cells of in vivo primed obese mice less active than those of lean controls, but also their number per spleen was significantly decreased. Such a quantitive difference was no longer found in adrenalectomised animals, but the qualitative difference in spleen cell cytotoxic activity still occurred. This suggests that adrenocortical hyperfunction may affect immune function in obese mice, without necessarily being the only factor in the in vivo environment of obese mouse spleen cells capable of depressing cellular immune reactivity.


Implications of aldose reductase in cataracts in human diabetes. Cataracts removed intracapsularly by cryoprobe technique from human diabetics were analyzed for sugars and polyols by gas liquid chromatography. The contents of sorbitol and fructose of lenses followed blood glucose levels at least up to 250 mg/dl. Studies indicate that human lens is capable of synthesizing substantial amounts of polyol pathway metabolites given exposure to high glucose levels such as are prevalent in diabetes. The synthesis of sorbitol was found to be susceptible to quercitrin, an inhibitor of aldose reductase. The implications of these findings in the formation of cataracts in diabetic individuals have been discussed.


Low-energy, Q-switched ruby laser iridotomies in Macaca mulatta. Laser iridotomies have been pursued as a means of performing anterior segments surgery as a virtually noninvasive procedure. An ideal single laser pulse technique has been elusive. In this study, iridotomies in rhesus monkeys were produced with a single exposure to a Q-switched ruby laser pulse. The iridotomy formation was accompanied by acoustic wave generation, bubble formation, and explosive tissue disruption, evidence of a nonlinear laser-iris interaction. The average energies at which these iridotomies were produced ranged between 18 and 48 mJ, some of the lowest energies reported for a laser iridotomy. Corneal changes were observed both at the epithelium and at the endothelium in some, but not all, of the eyes exposed. The epithelial changes morphologically resembled nonlinear damage reported for transparent solids. Damage to physical materials has been attributed to stimulated Brillouin scattering, a mechanism that may also play a role at the cornea. Consideration of such phenomena should be a part of the clinical evaluation prior to exposure of a cornea to high-power laser pulses. Although the endothelial change was more difficult to analyze, a shock-wave effect could not be discounted.


Stimulation of keratocyte metabolism by products of lymphoid cells. Products of leukocytes were found to activate cultures of keratocytes, manifested by the increased incorporation of thymidine or leucine. The keratocyte activation capacity crosses the species barriers between rabbit, monkey, and human. The level of secreted keratocyte-activating factor(s) (KAF) depends on the stimulation of the leukocytes. Thus unstimulated rabbit leukocytes produced very little or no KAF, whereas significant levels were produced by leukocytes stimulated with lipopolysaccharide (LPS) or concanavalin A. High levels of KAF were also found in supernatants of human mononuclear leukocytes stimulated by LPS. The effects of the activated leukocyte supernatants on keratocyte metabolism resembled the increased metabolic activity induced by the fibroblast and epidermal growth factors. The relationship between KAF and a possible modulating role of the products of lymphoid cells on corneal wound healing is suggested.


Health expenditure under multiple-priority pressures. A case study of Israel. Since Israel's inception, it has faced many problems that have a high priority in the national consciousness. Problems of defense and national security are outstanding among these, and problems such as absorption of immigrants, education, land reclamation and water supply are next. This creates heavy pressure on scarce resources, and is reflected in the share of specific items in Government and national expenditures. An analysis of developments since the early 1950s shows how health expenditure has been given a high priority in the evaluation of the nation's needs on one hand, and has been influenced by defense needs and other pressing requests on the other hand. It may be concluded that health receives a fair share of the nation's resources, but, in view of the strained economic situation in the country, careful planning for future health services in coordination, cooperation and integration with other welfare services is essential.


Preservation and storage of pathogenic Neisseria. Pathogenic species of Neisseria, especially N. gonorrhoeae are notoriously difficult to maintain in a viable state for long storage periods. Frequent transfer of cultures on artificial media and storage at 4 to 8 C quickly leads to loss of viability and virulence and to changes in antigenicity. Freeze-drying has been reported as a convenient method for long-term storage of these organisms; however, many laboratories are not properly equipped to use this method. A simple, inexpensive, and efficient procedure is described for storage of N. gonorrhoeae or N. meningitidis for 12 and 18 months, respectively. Results of this work may provide the basis for an efficient method of transporting these organisms over long distances via mail.


The dietary treatment of hepatic glycogen synthetase deficiency. A child with glycogen synthetase deficiency has been treated for one year by more frequent feeding with a diet lower in carbohydrate and higher in protein than her previous diet. This treatment virtually abolished the overnight hyperketonaemia and daytime hyperglycaemia which occurred before treatment. On the new dietary regime the child has had no clinical symptoms of hypoglycaemia and her growth velocity has increased dramatically.


Lack of response to vitamin D therapy in a patient with hypoparathyroidism under anticonvulsant drugs. Case report of a 16-year-old boy suffering from idiopathic hypoparathyroidism with severe seizures who erroneously and unsuccessfully had received anticonvulsive therapy for 12 years. When the correct diagnosis was made, treatment with massive doses of vitamin D2 was started. However, this treatment became effective only after the anticonvulsive therapy was discontinued; the serum calcium, phosphorus and the plasma 25-hydroxycholecalciferol returned to normal values, and the seizures stopped.


Non-Mendelian female sterility in Drosophila melanogaster: influence of ageing and thermic treatments. I. Evidence for a partly inheritable effect of these two factors. Crosses between certain Drosophila melanogaster strains may give rise to female sterility of non-Mendelian determination. Reduced fertility is observed in F1 females, known as SF females, from crosses between females of "reactive" strains and males of "inducer" strains. The extent of this reduction of fertility depends on the strains which are used in the cross and on two non-genetic factors: age and temperature. The fertility of SF females increases with ageing. Also, exposing them for a short period to a high temperature (29 degrees C) either increases or decreases the probability of hatching of the eggs according to the stage of oogenesis at which the heat treatment is applied. A very striking point is that qualitatively quite similar, though attenuated, effects are observed when the two factors (ageing and temperature) are applied not directly to SF females, but to their maternal ancestors: mothers and grandmothers.


[The expanding skull fracture (author's transl)]. An expanding skull fracture with exceptional extension into the frontal sinus, the orbit and the ethmoidal cells is discussed. A 12 year old boy fell down the stairs at the age of 8 years and fractured his left frontal bone with extension to the base of the skull. He was seen because of a swelling of the scalp, an intermittent orbital oedema and convulsions. At neurosurgery a large gaping fracture of the frontal bone was found and this was interposed with fibrous tissue and relatively denatured cerebral tissue. The management both neurosurgical and rhinological is discussed.


The development of the rods in the tapetal cells of the cat. The development of tapetal rods in the cat was studied ultrastructurally and cytochemically (DOPA reaction). The tapetum lucidum of fetuses, newborns, 3 weeks and 1 month old kittens was considered. The DOPA reaction showed that rod formation involved similar enzymic mechanisms to those concerned in the construction of melanosomes.


Direct comparison of the subtilisin-like intracellular protease of Bacillus licheniformis with the homologous enzymes of Bacillus subtilis. Intracellular serine proteases of Bacillus licheniformis and Bacillus subtilis are closely related.


Increased levels of dihydrofolate reductase in rifampin-resistant mutants of Bacillus subtilis. Several independent, spontaneous rifampin-resistant mutants of Bacillus subtilis were isolated and found to have an increased resistance to trimethoprim, an inhibitor of dihydrofolate reductase. This increased resistance in the rif mutants was the result of a specific threefold increase in the activity of dihydrofolate reductase, since six other enzymes examined remained unchanged. This increased level of dihydrofolate reductase and the trimethoprim resistance were cotransformed (100%) with the rif marker. These results suggest that the RNA polymerase is altered in its recognition of the gene that specifies dihydrofolate reductase.


Two forms of nitrogenase from the photosynthetic bacterium Rhodospirillum rubrum. Acetylene reduction by nitrogenase from Rhodospirillum rubrum, unlike that by other nitrogenases, was recently found by other investigators to require an activation of the iron protein of nitrogenase by an activating system comprising a chromatophore membrane component, adenosine 5'-triphosphate (ATP), and divalent metal ions. In an extension of this work, we observed that the same activating system was also required for nitrogenase-linked H(2) evolution. However, we found that, depending on their nitrogen nutrition regime, R. rubrum cells produced two forms of nitrogenase that differed in their Fe protein components. Cells whose nitrogen supply was totally exhausted before harvest yielded predominantly a form of nitrogenase (A) whose enzymatic activity was not governed by the activating system, whereas cells supplied up to harvest time with N(2) or glutamate yielded predominantly a form of nitrogenase (R) whose enzymatic activity was regulated by the activating system. An unexpected finding was the rapid (less than 10 min in some cases) intracellular conversion of nitrogenase A to nitrogenase R brought about by the addition to nitrogen-starved cells of glutamine, asparagine, or, particularly, ammonia. This finding suggests that mechanisms other than de novo protein synthesis were involved in the conversion of nitrogenase A to the R form. The molecular weights of the Fe protein and Mo-Fe protein components from nitrogenases A and R were the same. However, nitrogenase A appeared to be larger in size, because it had more Fe protein units per Mo-Fe protein than did nitrogenase R. A distinguishing property of the Fe protein from nitrogenase R was its ATP requirement. When combined with the Mo-Fe protein (from either nitrogenase A or nitrogenase R), the R form of Fe protein required a lower ATP concentration but bound or utilized more ATP molecules during acetylene reduction than did the A form of Fe protein. No differences between the Fe proteins from the two forms of nitrogenase were found in the electron paramagnetic resonance spectrum, midpoint oxidation-reduction potential, or sensitivity to iron chelators.


Precursor-product relationship of intracellular and extracellular lipoteichoic acids of Streptococcus faecium. Exponential biosynthesis and excretion of lipoteichoic acid (LTA) during the exponential phase of growth, and continued synthesis and excretion during valine starvation of Streptococcus faecium (S. faecalis ATCC 9790), were shown. During exponential growth, extracellular LTA (LTAx) accounted for approximately 13% of the total LTA in cultures, whereas during valine starvation, this percentage increased to approximately 60% within 4 h. LTAx was present in a low-molecular-weight, apparently deacylated form, whereas intracellular (LTAi) was present primarily in an apparently high-molecular-weight, acylated and micellar form. Experiments utilizing chases of either fully equilibrated or short pulses of [14C]- or [3H]glycerol were used to demonstrate that LTAx was derived directly from LTAi.


Influence of the lactose plasmid on the metabolism of galactose by Streptococcus lactis. Streptococcus lactis strain DR1251 was capable of growth on lactose and galactose with generation times, at 30 degrees C, of 42 and 52 min, respectively. Phosphoenolpyruvate-dependent phosphotransferase activity for lactose and galactose was induced during growth on either substrate. This activity had an apparent K(m) of 5 x 10(-5) M for lactose and 2 x 10(-2) M for galactose. beta-d-Phosphogalactoside galactohydrolase activity was synthesized constitutively by these cells. Strain DR1251 lost the ability to grow on lactose at a high frequency when incubated at 37 degrees C with glucose as the growth substrate. Loss of ability to metabolize lactose was accompanied by the loss of a 32-megadalton plasmid, pDR(1), and Lac(-) isolates did not revert to a Lac(+) phenotype. Lac(-) strains were able to grow on galactose but with a longer generation time. Galactose-grown Lac(-) strains were deficient in beta-d-phosphogalactoside galactohydrolase activity and phosphoenolpyruvate phosphotransferase activity for both lactose and galactose. There was also a shift from a predominantly homolactic to a heterolactic fermentation and a fivefold increase in galactokinase activity, relative to the Lac(+) parent strain grown on galactose. These results suggest that S. lactis strain DR1251 metabolizes galactose primarily via the tagatose-6-phosphate pathway, using a lactose phosphoenolpyruvate phosphotransferase activity to transport this substrate into the cell. Lac(-) derivatives of strain DR1251, deficient in the lactose phosphoenolpyruvate phosphotransferase activity, appeared to utilize galactose via the Leloir pathway.


Peptidoglycan turnover during growth of a Bacillus megaterium Dap- Lys- mutant. The aim of this study was to ascertain whether or not the absence of cell wall growth zones, deduced from the analysis of autoradiographs of DL-[3H]mesodiaminopimelic acid pulse-labeled cells of a Dap- Lys- mutant of Bacillus megaterium, was due to a high peptidoglycan turnover. Turnover was determined in very precise experimental conditions because two kinds of turnover occurred: a low, acid-soluble turnover and a high, acid-insoluble one. The latter was detected during a chase in the culture medium when bacteria were centrifuged before treatment with trichloroacetic acid. Otherwise the acid-insoluble released material precipitated with the bacteria. In the electron microscope this material presented a globular structure and contained both peptidoglycan and teichoic acid. The acid-insoluble turnover was mainly produced by a lytic acitivity that was released into the culture medium. This thermolabile activity was not due to cell lysis. It was implicated in septum cleavage and in the detachment of wall fragments from the cell surface, but did not seem indispensable for cell elongation. The acid-soluble turnover was much weaker and seemed to be indispensable for cell elongation.


Separation of inner and outer membranes of Rickettsia prowazeki and characterization of their polypeptide compositions. Rickettsia prowazeki were disrupted in a French pressure cell and fractionated into soluble (cytoplasm) and envelope fractions. The envelope contained 25% of the cell protein, with the cytoplasm containing 75%. Upon density gradient centrifugation, the envelope fraction separated into a heavy band (1.23 g/cm3) and a lighter band (1.19 g/cm3). The heavy band had a high content of 2-keto-3-deoxyoctulosonic acid, a marker for bacterial lipopolysaccharide, but had no succinic dehydrogenase, a marker for cytoplasmic membrane activity, and therefore represented outer membrane. The lighter band exhibited a high succinate dehydrogenase activity, and thus contained inner (cytoplasmic) membrane. Outer membrane purified by this method was less than 5% contaiminated by cytoplasmic membrane; however, inner membrane from the gradient was as much as 30% contaminated by outer membrane. The protein composition of each cellular fraction was characterized by sodium dodecyl sulfate--polyacrylamide gel electrophoresis. The outer membrane contained four major proteins, which were also major proteins of the whole cell. The cytoplasmic membrane and soluble cytoplasm exhibited a more complex pattern on gels.


Neurotoxicity with combined use of lithium and thioridazine. Four cases on a combination of lithium and thioridazine developed severe neurotoxic symptoms, e.g., delirium, seizures, encephalopathy and grossly abnormal EEG's. All patients had been on lithium without thioridazine prior to and after the neurotoxic episode with no complications or EEG changes. The possible risk of combining lithium and neuroleptics (thioridazine) is discussed and repeated EEGs are recommended as a preventive measure.


Isolation, homogeneity, and properties of core particle from pyocin R1. By a mild alkaline treatment of pyocin R1, the core particle was released from the contracted sheath. After sucrose density-gradient centrifugation, core-rich fractions were treated with anti-sheath serum and by a second density-gradient centrifugation, purified core particles were isolated. Homogeneity of the preparation was confirmed by observation under the electron microscope, immuno-precipitation reaction, and sucrose density-gradient centrifugation. The core particle exhibited a sedimentation coefficient of 37S. The quaternary structure of the core consists of a single kind of subunit protein with a molecular weight of 18,000. No contamination by other proteins was detected by SDS-disc electrophoreses. Amino acid analysis revealed that the core is rich in glycine, alanine, valine, leucine, aspartic acid (or asparagine), glutamic acid (or glutamine), and serine. This amino acid composition bears some resemblance to that of T-even bacteriophage tail-core.


Studies on cellulases of a phytopathogenic fungus, Pyricularia oryzae Cavara. III. Multiplicity of beta-glucosidase, and purification and properties of a second component. To determine the relationship between the induction patterns of three components of beta-glucosidase of Pyricularia oryzae and carbon sources in the growth medium, various culture conditions were examined. Avicel, hydroxyethylcellulose and methyl-beta-D-glucoside as the carbon source induced both beta-glucosidase components, GB-1 and GB-2, whereas cellobiose and gentiobiose induced only one component, GB-1. Thus, these two components were induced independently and hence thought to be isozymes. The GB-2 was purified to homogeneity by ion exchange and gel filtration chromatographies from two different cultures on methyl-beta-D-glucoside and Avicel. The specific activity of GB-2 when salicin was used as substrate was approximately 5.9 mg glucose/min/mg protein. GB-2 was found to be an oligomeric glycoprotein, which consisted of two subunits with molecular weight of approximately 120,000, comprising a relatively large number of acidic amino acids and mannose, as is the case with GB-1. These two isozymes were clearly different in thermostability, GB-2 being more thermolabile than GB-1. However, the same carboxyl group (pKa 4.2--4.8) was found to be strongly implicated in the formation and dissociation of the enzyme-substrate complex for both of the enzymes, from the analysis of kinetic parameters as a function of pH.


The role of concanavalin A dissociation on positive cooperativity of binding with native and fixed erythrocytes. Positive cooperativity demonstrated by Scatchard plot analysis of concanavalin A (con A) binding was found with native or glutaraldehyde-fixed erythrocytes. This suggests that factors other than membrane changes might be involved in the apparent increase in receptor binding affinity with increasing site occupancy. The elution pattern of 125I-con A chromatographed on Bio-Gel P-150 with decreasing concentration showed a drop in average molecular weight compatible with con A dissociation to dimers, protomers, and protomer fragments. Similarly, the per cent of 125I-con A specifically binding to Sephadex G-75 fell with decreasing concentration of con A applied. The inclusion of unlabeled carrier con A suppressed the dissociation of labeled con A in Bio-Gel P-150 and increased the per cent binding to Sephadex G-75. Both labeled and unlabeled con A were multibanded in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As previously reported, the three major bands are consistent with intact protomer (approximately 25,000 daltons) and two fragments (approximately 13,000 and 10,000 daltons) with minor bands representing undissociated species. These observations indicate that there is a concentration-dependent association of Con A subunits which contribute to the observed positive cooperativity of con A binding to erythrocytes.


Identification of a novel RNA molecule in a new RNA processing mutant of Escherichia coli which contains 5 S rRNA sequences. A temperature-sensitive mutant of Escherichia coli at the nonpermissive temperature fails to produce normal levels of 5 S rRNA. Instead, a number of larger RNA molecules are accumulated. One of these molecules, a 9 S RNA, contains 5 S rRNA sequences. When the strain is shifted from a nonpermissive to a permissive temperature, radioactive label is lost from the 9 S RNA and appears in 5 S rRNA. The identification of this 5 S rRNA-containing molecule indicates the participation of a new processing ribonuclease (RNase E) in the maturation of rRNA in E. coli. The 9 S RNA was not detected in a wild type strain, indicating that the processing step(s) involved in the formation of 5 S rRNA might be performed before the growing rRNA transcript is terminated.


B-type cytochromes in plasma membranes isolated from rat liver, in comparison with those of endomembranes. Fractions of plasma membranes, Golgi apparatus, endoplasmic reticulum (ER), and nuclear envelope were isolated from rat liver and were characterized by electron microsocpe and biochemical methods. The purity of the fractions was controlled by morphometry and by marker enzyme activities. Amounts of cytochromes b5, P-450, and P-420 were measured, as well as the NADPH- and NADPH-cytochrome c reductase activities. The pigments of the microsomal electron transport system were found in all membrane fractions in relatively high amounts, thus excluding an origin by microsomal contamination. Purified preparations of plasma membrane and Golgi apparatus contained approximately 30% of the cytochrome b5 and cytochrome P-450 + P-420 found in ER membranes. Plasma membranes were also characterized by a high ratio of P-420/450. Degradation of cytochromes P-450 and P-420 was relatively rapid in all fractions, except in the ER. Cytochrome b5 extracted from plasma membranes was spectrophotometrically and enzymatically indistinguishable from ER cytochrome b5. However, immunnlogical characterization with rabbit antibodies against the trypsin-resistant core of microsomal cytochrome b5 showed the presence of at least two types of cytochrome b5 in ER membranes, in contrast to the plasma membranes in which only one of these components was detected. This immunological differentiation also demonstrates that the plasma membrane-bound cytochrome b5 is endogenous to this membrane and does not reflect contamination by ER elements. We conclude that cytochromes b5, P-450, and P-420 are not confined only to ER and nuclear membranes but also occur in signficant amounts in Golgi apparatus and plasma membranes. The findings are discussed in relation to observations of similar redox components in Golgi apparatus, secretory vesicles, and plasma membranes of other cells.


Regulation of RNA synthesis in Tetrahymena pyriformis: secretion of regulatory factors. The rate of ribosomal RNA synthesis varies greatly with the population density in both exponentially and synchronously growing populations of Tetrahymena pyriformis. Shortly after inoculation of the population - at relatively low cell densities - a gene-dose effect dominates the picture, and a doubling in the gene number is immediately followed by a doubling in the rate of RNA synthesis. However, also other mechanisms are controlling the rate of RNA synthesis. Generally one finds high rates of RNA synthesis in the lag phase of newly inoculated cells, decreasing rate of RNA synthesis during most of the exponential growth phase and very low rate of synthesis in stationary phase cells. We now have results which show that the repression of RNA synthesis in densely populated cultures is caused by a dialysable factor, which is secreted by the cells. If cells are inoculated on a medium which contains this factor the high initial rate of RNA synthesis normally observed is prevented, but the cells multiply and grow with normal generation time until normal stationary-phase population densities are reached.