Pulmonary anaphylaxis and the kallikrein-kinin system. The kallikrein-kinin system has been thought to participate in the pathogenesis of anaphylaxis. Kallikrein, released from lungs, has been postulated to contribute to cardiovascular collapse. Further to test the hypothesis, we examined for the occurrence of a kallikrein-like enzyme in guinea pig lungs and examined for release of such an enzyme by isolated, perfused lungs of guinea pig sensitized to and challenged with egg albumin. In addition, we treated guinea pigs with the bradykinin potentiating agents, BPP9a and SQ 14,225. In parallel experiments, we examined for effects of non-steroidal anti-inflammatory agents on the supposition that prostaglandin-related substances may mediate or modulate actions of kinins during anaphylaxis. A plasma kallikrein-like enzyme was found in lung homogenates and occurred in concentrations greater than that of plasma itself. Similarly, a store of kininogen occurs in lungs. However, using a sensitive radioassay for kallikrein-like enzymes, we were unable to confirm that antigenic challenge of sensitized lungs causes the release of enzyme into pulmonary venous effluent. Further, we were unable to modify the acute course of anaphylaxis by pretreatment of guinea pigs with bradykinin potentiating agents. However, indomethacin and aspirin at 20-40 mg/kg were found to greatly increase the severity of pulmonary anaphylaxis in terms of increased resistance to ventilation and increased numbers of lung hemorrhages. Paradoxically, aspirin or sodium salicylate at 80-100 mg/kg prevents the characteristic rise of insufflation pressure and the formation of lung hemorrhages.


Altered urinary excretion of human kininase activity in acute myocardial infarction. This study concerns the determination of levels of human urinary kininase excretion in acute myocardial infarction (AMI). The results obtained by a biological method show that there is a significant reduction of the enzymatic activity in patients affected by AMI in comparison with normals (6.4 +/- 0.4 ng of destroyed bradykinin/min. versus 164.4 +/- 31.4 ng; P less than 0.001), while urinary kallikrein excretion was close to normal values.


The role of the kidney in vitamin D metabolism. The kidney should be regarded as an organ of central importance in the regulation of calcium metabolism. Its secretion product, 1 alpha,25(OH)2D3, is regulated by several factors, including the calcium and phosphorus content of the diet, parathyroid hormone and the level of 1 alpha,25(OH)2D3 itself. The main occasions in health where 1 alpha,25(OH)2D3 secretion is enhanced are during growth and reproduction. In these situations the secretion of this new renal hormone is under at least partial control of growth hormone and prolactin.


Metabolic and cellular activity of vitamin D. Most of the effects of vitamin D on calcium and phosphorus homeostasis result from an increase in intestinal absorption of calcium and phosphorus and from bone mobilization. These effects are due to 1,25(OH)2D3, one of the renal vitamin D metabolites. Besides ionic mobilization, vitamin D may exert additional effects on bone which depend on metabolites other than 1,25(OH)2D3. Kidneys, parathyroid glands and muscle are probably target organs for vitamin D. The metabolite(s) responsible for these actions remain unknown.


Pathways of arachidonic acid metabolism and modulation by drugs. The AA cascade in addition to producing various oxygenation products concomitantly yields an oxygen radical. This oxidant selectively deactivates the cyclooxygenase, peroxidase and PGI2 synthetase components whereas TXA2 synthetase is resistant to this oxidant. On the basis of available data it is not possible to assign an inflammatory inducing role to this oxidant on the basis of these actions. Other loci of action must be involved. Among the possibilities are the elements involved in the release of arachidonic acid. Studies relating to this possibility are examined in this report.


[Action of vitamin D and E on the erythrocyte membrane. II. Morphological changes in the erythrocytes studied by transmission electron microscopy]. Man, rabbit, guinea-pig erythrocytes treated "in vitro" with high doses of cholecalcipherol or alpha-tocopherol release K+ and haemoglobin and show ultrastructural damages. These damages consist in a change of the red cells profile as well as in the desappearance of the glycocalix, in alterations of the membrane and of the cortical cytoplasm structure. These ultrastructural alterations are of a different degree, depending on the animal species; they partly resemble the erythrocyte changes induced by retinol. Our results are in partial agreement with the Dingle and Lucy's hypothesis on the common action of liposoluble vitamines on the erythrocyte membrane.


Application of case series review results to the evaluation of individual cases in diagnostic radiology. Probability theory provides a simple method for physicians to use their "intellectual linkages" to their past clinical experience in making current diagnoses. Only a pencil and paper are required for making a few likelihood calculations. To illustrate this method, evaluation was done of a new diagnostic sign (presence of knee ossification centers) for differentiating rubella from cytomegalovirus infection in young infants. Two practical questions can be answered by use of this method for calculating probabilities: (1) How certain can one be about either diagnosis when centers are present or absent? (2) How can other radiologists apply these results to their individual cases?


Relationship between extent of coronary artery disease and correlative risk factors. An analysis was made of correlative factors which might be related to the angiographically measured extent of coronary artery disease in 140 patients. All patients presented with clinically important chest pain. Thirty-three had a normal coronary arteriogram. The extent of the atheromatous process was measured precisely at angiography by three different techniques. A coronary score, based on the percentage of luminal narrowing, was found to be best suited for the analysis. The most important contributory factors to the severity of atherosclerosis was duration of clinical history, number of previous myocardial infarctions, and male sex, but more specifically elevation of serum cholesterol and diabetes mellitus. Cigarette smoking, obesity, hypertension, a family history of atherosclerosis, and elevated serum triglycerides had a positive influence but this was not statistically significant.


Role of calcium and cyclic adenosine 3':5' monophosphate in regulating smooth muscle contraction. Mechanisms of excitation-contraction coupling in smooth muscle. Caclium initiates smooth muscle contraction by activating an enzyme, myosin light chain kinase. This enzyme catalyzes the transfer of phosphate from adenosine triphosphate to the 20,000 dalton light chain of myosin. In its phosphorylated form myosin interacts with actin to produce muscle contraction. The mechanism by which calcium activates myosin kinase requires (1) the binding of calcium to a 16,500 dalton calcium-binding protein (calmodulin), and (2) the binding of calmodulin-calcium to a 125,000 dalton catalytic subunit. This two protein complex is the active form of myosin light chain kinase. Smooth muscle relaxation is mediated by cyclic adenosine 3':5' monophosphate (cyclic AMP). One nechanism by which the latter may exert a direct effect on actin-myosin interaction is through the activation of a cyclic AMP-dependent protein kinase that can phosphorylate the 125,000 dalton component of myosin light chain kinase. Phosphorylation of myosin light chain kinase decreases the activity of the enzyme, thus favoring the unphosphorylated form of myosin, which cannot interact with actin to produce smooth muscle contraction.


Abnormal technetium-99m pyrophosphate images in unstable angina: ischemia versus infarction? There is controversy concerning the specificity of myocardial infarct imaging with technetium-99m pyrophosphate due to the high frequency of false positive images, especially in patients with unstable angina. In this study technetium-99m pyrophosphate images were compared with frequent determinations of plasma creatine kinase, MB isoenzyme (MB CK) activity in 116 patients admitted with the diagnosis of unstable angina. It was hypothesized that frequent measurement of MB CK activity, a sensitive and specific marker for myocardial necrosis, using sensitive assay techniques would detect small amounts of myocardial necrosis which might have been unrecognized by conventional clinical methods. The scintigraphic results and isoenzyme determinations agreed in 88 percent of patients; both tests were normal in 69 percent and both were abnormal, indicating acute myocardial infarcation, in 19 percent of patients. In the remaining 14 patients (12 percent), the scans were abnormal, but MB CK activity was normal. In five of these patients (4 percent), abnormal scintigrams presumably reflected persistent scan positivity after previous myocardial infarction. Only the remaining nine patients (8 percent) could be classified as having unexplained false positive scans, a frequency substantially less than that reported by other investigators who based the diagnosis of myocardial infarction on conventional clinical criteria. These results suggest that abnormal technetium-99m pyrophosphate images in patients with unstable angina generally indicate myocardial necrosis.


Acute lymphoblastic leukemia, hand-mirror variant. A detailed ultrastructural study. The hand-mirror cell in acute lymphoblastic leukemia may have important immunologic, pathogenic, and prognostic implications. To learn more about this cell, a detailed ultrastructural analysis was performed. Fifty electronmicrographs of lymphoblasts from an untreated patient with acute lymphoblastic leukemia with numerous hand-mirror cells in the bone marrow were compared with 60 electronmicrographs of lymphoblasts from six patients with classic acute lymphoblastic leukemia. The unique qualitative ultrastructural features of the hand-mirror cells were the presence of undamaged mitochondria and uropods (handles) with terminal microspikes containing circular organized arrangement of 50-A microfilaments. Quantitative differences between hand-mirror cells and lymphoblasts were observed in nuclear perimeters (P less than 0.0001), nuclear lengths (P less than 0.001), cytoplasmic lengths (P less than 0.0002), nuclear-cytoplasmic length ratios (P less than 0.0001), and numbers of mitochondria (P less than 0.002). These findings indicate that hand-mirror cells contain ultrastructural components that are related to cell motility and the immunologic process. These results are significant in that the hand-mirror cells may be associated with an immunologic mechanism that is involved in leukemogenesis.


Cytomegalovirus encephalitis associated with thymoma and immunoglobulin deficiency. The clinical and immunologic findings in an elderly woman with thymoma and immunoglobulin deficiency in whom cytomegalovirus (CMV) encephalitis developed are described. The patient had absent serum immunoglobulins and no circulating immunoglobulin-bearing lymphocytes. Complement receptor-bearing lymphocytes were present in the peripheral blood, and circulating T lymphocyte numbers were within normal limits. She was anergic to a battery of skin test antigens, and her lymphocytes in vitro showed a selective unresponsiveness to CMV antigen while responding normally to phytohemagglutinin and streptokinase. The course of the encephalitis was progressive with quadriplegia, aphasia and coma developing within six months of onset of symptoms. This is the fifth reported case documenting an association between CMV infection and the syndrome of thymoma with immunoglobulin deficiency, but the first report of fatal CMV encephalitis in a patient with thymoma and immunoglobulin deficiency.


Estrogen-binding proteins in the human postmenopausal uterus. Two intracellular high-affinity, low-capacity estrogen-binding proteins, which have the characteristics of receptors, with equilibrium dissociation constants of 10(-10)M and 10(-9)M, have been observed in the human uterus. The higher-affinity protein (10(-10)M) appears to play the main role in activating end-organ response to estrogen stimulation. The role of the lower-affinity protein (10(-9)M) is uncertain. In a human postmenopausal uterine system without estrogen stimulation, Scatchard analysis of uterine cytosol partially purified by ammonium sulfate fractionation and incubated at 4 degrees C for 18 hours revealed only the higher-affinity receptor component (10(-10)M). In a post menopausal uterine system with estrogen priming (and in the premenopausal uterus) both the high- and low-affinity components were observed. Competition studies indicated that the receptors were specific for estradiol. The clinical significance of these findings is discussed.


Stigma and state hospital patients.

Stigma and state hospital patients. This study set out to measure the degree of social stigma experienced by discharged mental hospital patients and the extent to which these patients were viewed as a burden by family and friends. Predictors of stigma included social class and demographic factors, posthospital situation, emotional functioning, and use of aftercare facilities. However, available measures were able to account for little of the variance in stigma, and further research is suggested to establish more reliable predictors of mental patient stigmatization.


Transport and metabolism of fructose in fat cells of normal and hypophysectomized rats. We have shown earlier that the glucose carrier of fat cells of hypophysectomized rats operates at maximal speed in the basal state and that these cells are insensitive to insulin. Here we characterize fructose transport and metabolism in adipocytes of normal and hypophysectomized rats. 3-O-methylglucose inhibits insulin-stimulated incorporation of [U-14C]fructose into fat cells and has no significant effect on basal fructose metabolism. In contrast, 2-deoxyglucose inhibits both basal and insulin-stimulated fructose incorporation. Insulin enhances fructose incorporation in normal adipocytes only in the absence of 3-O-methylglucose. In hypophysectomized rats, incorporation of glucose and of fructose is decreased and insensitive to insulin. In contrast to the glucose carrier, the specific fructose carrier appears to be insensitive to insulin and to continue to operate with unchanged characteristics after hypophysectomy. Thus, hypophysectomy leads to a specific alteration of the insulin-sensitive glucose carrier, whereas the insulin-insensitive fructose carrier remains unaltered.


Possible role of cyclic GMP in stimulus-secretion coupling by salt gland of the duck. Stimulation of salt galnd secretion in domestic ducks in vivo increased the cyclic GMP concentration of the tissue, but had no effect on cyclic AMP levels. Methacholine, which is known to stimulate sodium transport by the glands both in vivo and in vitro, stimulated ouabain-sensitive respiration in salt gland slices. Cyclic GMP stimulated ouabain-sensitive respiration to the same extent as methacholine. Guanylate cyclase stimulators, hydroxylamine and sodium azide, also stimulated ouabain-sensitive respiration. The stimulation of ouabain-sensitive respiration by methacholine was blocked either by atropine or by removal of calcium from the incubation medium. The stimulation of ouabain-sensitive respiration by cyclic GMP still occurred in the absence of calcium. The above observations seem to indicate that cyclic GMP acts as a tertiary link in the process of stimulus-secretion coupling in the tissue.


Effect of Amytal on metabolism of perfused rat heart: relationship between glycolysis and oxidative phosphorylation. In perfused rat hearts, infusion of increasing concentration of Amytal caused progressive inhibition of respiration and increase in glycolytic activity. At maximal inhibition of respiration, with glucose as the substrate, glycolysis provided about 60% of the total ATP produced. The myocardial content of ATP remained constant irrespective of the infused Amytal concentration but [CrP]/[Cr] and [ATP]/[ADP]f[Pi] progressively decreased. Changes in the concentrations of glycolytic intermediates were observed, the most pronounced of which were increases in fructose 1,6-diphosphate and lactate contents and a decrease in the pyruvate level. Myocardial levels of oxaloacetate, malate, and alanine were elevated and so was alanine release from the tissue. Substitution of glucose with pyruvate caused a large increase in the concentrations of the tricarboxylic acid cycle intermediates and consequent accumulation of reducing equivalents in the mitochrondria. With the latter substrate, in the presence of Amytal, the rates of mitochondrial ATP production were higher than those with glucose as the substrate. The metabolic picture of the Amytal block resembles biochemical manifestations of human myopathies of mitochondrial origin, and therefore Amytal inhibition is a convenient model system for exploration of intermediary metabolism in these defects.


Role of Ca in isoproterenol-induced increases in cAMP levels in rat uterus. The influence of alteration in the Ca2+ environment of the tissue on isoproterenol-induced increases in cAMP levels and relaxation was studied in rat uterus. In muscles depolarized with 47.5 mM K+ (with or without Na+), the ability of isoproterenol to increase cAMP levels and to produce relaxation was found to be inversely related to external calcium concentration. The pretreatment of the muscle with D600 or EGTA restored the cAMP response to isoproterenol in the depolarized uterus to a level observed in nondepolarized muscle. The study with Ro 20-1724, a phosphodiesterase (PDE) inhibitor indicated that the failure of isoproterenol to elevate cAMP levels in the depolarized uterus could not be related to the activation of PDE by Ca2+. The exposure of rat uterus to a zero-Ca2+ solution accentuated the increases in cAMP levels produced by isoproterenol. These results have raised the question of a possible regulatory role of Ca2+ in beta-adrenoceptor-induced increases in cAMP levels in uterine smooth muscle.


Hyperglucagonemia and alpha-adrenergic receptor in acute hypoxia. The plasma immunoreactive glucagon (IRG) response to hypoxia was studied in puppies. Three groups of paired experiments were performed. In group I, 8% O2:92% N2 ventilation (PaO2 20--30 torr) produced a rise in plasma IRG and glucose as well as hypotension and bradycardia. However, when group I was air ventilated (PaO2 greater than 70 torr) and given glucose infusions producing hyperglycemia of similar degree, plasma IRG was unchanged. Group II received alpha-adrenergic blockade (phenoxybenzamine). When made hypoxic, group II developed no significant IRG rise and less hyperglycemia than with hypoxia alone. Hypotension was more severe with hypoxia plus alpha-blockade. Phenoxybenzamine itself did not change plasma IRG or glucose during air breathing. Group III receivi developed hyperglucagonemia and hyperglycemia not significantly different from that with hypoxia alone. However, hypoxia-caused hypotension and bradycardia was more pronounced with beta-blockade. No change in plasma IRG or glucose occurred in group III animals breathing air. These data suggest that a) glucagon release is caused by acute oxygen deficiency, and b) the hypoxic response is largely adrenergically mediated with the major role played by the alpha-receptor.


Actions of histamine, secretin, and PGE2 on cyclic AMP production by isolated canine fundic mucosal cells. Cyclic AMP production was studied in isolated canine fundic gastric mucosal cells. Histamine, prostaglandin E2 (PGE2), and secretin increased cyclic AMP production by unenriched mucosal cells. In separated cell fractions, histamine stimulation of cyclic AMP production correlated with the parietal cell content of the fractions. Secretin in concentrations above 1 nM stimulated cyclic AMP production, and this effect correlated with the pepsinogen content of the separated cell fractions. At concentrations above 1 microM, PGE2 stimulated cyclic AMP production; this effect was found in all separated cell fractions and was not associated with any of the available cell markers. PGE2 stimulation of cyclic AMP production was, however, negatively correlated with the parietal cell content. Thus, histamine stimulated cyclic AMP production by parietal cells and secretin stimulated production of cyclic AMP by chief cells. PGE2 stimulation of cyclic AMP production could not be localized to a single cell type but occurred primarily in nonparietal cells.


Effect of sodium restriction on plasma renin activity and renin granules in rat kidney. The present study was carried out to procure detailed information on the relationship between chronic sodium restriction and renin content of kidneys at a subcellular level in the rat. Renin granules (RG) were separated by a discontinuous sucrose-density gradient (from 1.2 to 1.7 M) centrifugation. In control rats, RG were mainly recovered in the fractions corresponding to 1.5 M sucrose, whereas most of the mitochondria, lysosomes, and microsomes equilibrated in upper fractions. The RG fraction contained approximately 60% of total granular renin activity. Low sodium intake for 4 wk resulted in a 12.4-fold increase in plasma renin activity and led to a 2.6-fold increase in renin activity of the RG fraction. But in sodium-restricted rats there was no alteration in the distribution pattern of renin activity on sucrose-density gradients, indicating that there was no change in the density of RG. These results provide evidence for increased renin activity in storage granules following chronic sodium restriction.


Metabolism of totally ischemic excised dog heart. II. Interpretation of a computer model. Analysis of the ischemic dog heart preparation described in the preceding paper indicates that it is an analogue in slow motion of the tissue in the center of a cardiac infarct. It is respiring very slowly and not capable of performing mechanical work. Glycolysis starts up with both glucose and glycogen as inputs. Later hexokinase and to some extent phosphofructokinase become limiting owing to inhibitor accumulation or acidosis. Metabolism then results primarily from cAMP-driven glycogenolysis, largely limited by the glycogen debranching enzymes at later times, with accumultion not only of lactate and alpha-glycerophosphate but of glucose as well. Amino acid levels oscillate with time while fatty acids accumulate at late times. The elevation of cAMP at later times may involve disturbances in its metabolism as well as mechanisms such as adenosine accumulation that are more important in cardiac ischemia than in normal heart. The clinical implications of this behavior are discussed.


REM architecture changes in bipolar and unipolar depression. The authors compared total night sleep measures and REM sleep architecture values for normal control subjects (N = 36), unipolar depressed patients (N = 36), and bipolar depressed patients (N = 22). The unipolar and bipolar patients had significantly greater fragmentation of REM periods than control subjects, and bipolar patients showed greater fragmentation of REM periods than unipolar patients. In both the unipolar and bipolar samples, the duration of successive REM periods was related to the total number of REM periods during sleep.


Prognosis after resection of metachronous multiple bronchogenic carcinoma. Information on 55 cases of multiple metachronous lung cancer in which both tumors were resected was collected from published reports for comparison with a series of 214 single cancer cases treated by lobectomy in Philadelphia teaching hospitals. The two groups were similar with respect to age, extent of resection, and histologic type of cancer (the second lesion in the multiple cancer group). The life table method was used to assess survival in the multiple cancer group for 4 years after the second resection. Survival in the two groups was similar in the first 2 years, but at the end of the third and fourth years the survival rate for the multiple cancer group was only half that of the single cancer group. These differences were statistically significant. Prognosis 4 years after resection of a second metachronous bronchogenic carcinoma is poor.


Sleep in herniorrhaphy patients.

Sleep in herniorrhaphy patients. The nocturnal sleep patterns of 10 elective herniorrhaphy patients were documented by continuous, 8 hour polygraphic recordings from night 1 of hospitalization until discharge on day 4 or 5. Subjects showed a dramatic loss of REM sleep and stages III and IV and major increases in time awake and drowsy for the first 2 postoperative nights, with a gradual return toward normal sleep over the hospitalization period. The significance for healing of the loss of stages REM, III, and IV is discussed, especially with regard to hospital procedures and the use of medications.


[Bronchial granular cell tumor. Apropos of 2 cases]. The authors report two new cases of bronchial granular cell tumour, a site which remains relatively rare. One of these cases was studied from the point of view of histoenzymology and ultrastructure. The appearances seen were identical to those seen in the literature. They lead to the problem of the histogenetics of the lesion, various morphological factors favouring a Schwann cell origin, whilst the presence of "transitional" cells is more in favour of a mesenchymatous origin, or at least the role of mesenchymatous cells in the development of these lesions.


Live cytomegalovirus vaccination of renal transplant candidates. A preliminary trial. Significant morbidity and mortality are associated with primary cytomegalovirus infections in renal-transplant recipients. In the hope that immunity to cytomegalovirus could safely be established before transplantation, we vaccinated 12 seronegative renal-transplant candidates with the Towne 125 strain of live human cytomegalovirus. Before transplantation, there were no significant reactions except for erythema and induration at the site of inoculation. All vaccinees seroconverted, and the three patients tested acquired a cytomegalovirus-specific cellular immune response. Ten vaccinees underwent transplantation: Nine have completed at least 3 months of follow-up, and eight retain functioning allografts up to 1 year later. Although cytomegalovirus was isolated from six patients after transplantation, the restriction endonuclease patterns of the viral DNA of four of these isolates differed significantly from those of the vaccine strain. Therefore, it appears that the vaccine strain did not become latent in the host, at least in a form that could be reactivated.


Possible alpha-adrenergic involvement in nicotine induced alteration of spermatogenesis in rat. Nicotine caused a fall in primary spermatocyte number together with a retardation of spermatocyte to spermatid conversions process. These effects of nicotine were possibly secondary to increased activity of adrenomedullary catecholamines, since the antispermatogenic effect of nicotine was prevented by phentolamine, a known blocker of catecholamine action at the alpha-receptor level.


Asymptomatic glomerulonephritis after nonstreptococcal upper respiratory infections. Two hundred forty previously healthy military personnel with nonstreptococcal upper respiratory infections were prospectively studied to define the incidence and clinicopathologic characteristics of possible virus-associated glomerulonephritis. Nine patients without preceding streptococcal infection had erythrocyte casts on urinalysis and glomerulonephritis on biopsy. Of these nine, four had a reduction in total hemolytic complement and five had serologic evidence of infection with adenovirus, influenza A, or influenza B. Initial renal biopsy showed either focal or diffuse mesangial proliferation in all nine, with mesangial C3 deposits in six specimens. Repeat biopsy in three showed histologic improvement or loss of immunofluorescent staining, or both. Sequential creatinine clearances were reduced to 74 to 90 mL/min.1.73 m2 in five patients for the duration of follow-up. We conclude that nonstreptococcal upper respiratory infection is frequently associated with glomerulonephritis and that abnormal glomerular structure and decreased creatinine clearances may persist for at least 2 to 8 months.


Serotoninergic involvement in the regulation of circadian rhythms of hypothalamic-pituitary adrenal and prolactin axis. A detailed review of their own experiments and of data existing in the literature regarding the role played by 5HT in the control of circadian rhythms of HHAA and HHPA is given by the AA. In rats, a positive correlation between 5-HT content in the limbic system and plasma corticosterone circadian rhythm has been shown to exist by means of pharmacological and neurosurgical approaches. Lesions of the raphe area result in an abolition of diurnal periodicity of plasma steroids similar to that obtained after inhibition of 5-HT synthesis by means of PCPA. A modification of the amplitude of the plasma corticosterone circadian variations also occurs after more selective destruction of some nuclei (n. centralis superior and n. raphe dorsalis) which reduced 5-HT concentration both in the hypothalamus and the hippocampus. Isolation of the mediobasal hypothalamus by a frontal cut posterior to the optic chiasma abolishes the diurnal fluctuation of plasma corticosterone, indicating that the suprachiasmatic nucleus is a focal site for the 5-HT regulation of the phasic activity of the HHAA. Evidence in favour of 5-HT mechanisms in the limbic area as the biological clock regulating the circadian variations of the adrenal axis is also provided. Restoration of the diurnal rhythm of plasma corticosterone after its abolition could be explained with the existence of a small functional pool of brain 5-HT or with the development of receptor supersensitivity. Pharmacological and surgical manipulation on central 5-HT neurons suggest the existence of two distinct serotoninergic mechanisms, a limbic one and an intrahypothalamic one, which play a modulatory role in the circadian variations of plasma PRL. Also in this case, adaptive accessory brain structures can take over in restoring the circadian hormones periodicity in conditions of impairment of the main serotoninergic mechanisms.


The participation of monoaminergic central mechanisms on adenohypophyseal secretion. In acute and chronic experiments in rats has been studied the action of drugs influencing central neuro-transmitter on the functional activity of hypophysis-adrenal and hypophysis-gonadal system. Evidence has been obtained that central neurotransmitters participate in the control of ACTH-glucocorticoids secretion. However, there is no clear correlation between noradrenaline and serotonin level in the brain and the degree of reaction of pituitary-adrenal complex to stimuli. Pharmacological analysis of nervous regulation of gonadotropic hypophysis function has revealed the significance of adrenergic mediation in the mechanism of feedback action of sex hormones and in the mechanism of limbic influences on hypothalamic regulation of gonadotropin secretion.


Effect of drugs with addicting properties on brain catecholamines. The effect of morphine and amobarbital on catecholamine metabolism in different regions of the rat brain (cortex, hippocampus, striatum, hypothalamus and midbrain) was studied. The investigation showed that drugs of different chemical structures but with common addicting properties evoke the same type of changes in catecholamine metabolism which are expressed by an intensification of release and disintegration of norepinephrine and by a decrease in dopamine level.


Release of dopamine from striatal synaptosomes. The mechanisms of dopamine (DA) release central nerve endings have been investigated utilizing superfused rat striatal synaptosomes. Nomifensine was selected as a tool to discriminate between release mediated by the DA carrier and release occurring independently of the carrier. The following conclusions can be drawn from the results obtained: 1) Alterations of the sodium gradient across the synaptosomal membrane, induced by omission of extracellular Na+ or by ouabain, enhanced the release of 3H-DA from prelabeled synaptosomes. The release was blocked by nomifensine and therefore it was carrier-mediated. 2) The release of DA elicited by amphetamine and related phenylethylamines was nomifensine-sensitive, suggesting that the released DA existed from synaptosomes through the membrane carrier. 3) Depolarization of synaptosomes by high K+ triggered the release of both "newly taken up" and "newly synthesized" DA. 4) The calcium-dependent release of DA (induced by high K+, veratridine of by the ionophore A23187) was not affected by the carrier blocker nomifensine and may occur by an exocytic-like process. 5) The effects of apomorphine and neuroleptics on the stimulus-evoked release of DA do not support the existence of a presynaptic receptor-mediated inhibitory control of DA release identical to that described for noradrenaline.


[Isolated trigeminal neuropathy revealing the presence of Sharp's syndrome or MCTD (mixed connective tissue disease) (author's transl)]. Isolated neuropathies of 5th cranial nerves are rare apart from primary facial neuralgia. A case is reported, in which Raynaud's syndrome was also present, that enabled a diagnosis of mixed connective tissue disease to be established. The authors underline the importance of undertaking complete immunological investigations in order to decide whether there is a particular type of systemic disease present, which could be responsible for these special types of trigeminal nerve disorders.


Plasma lipids on chronic uraemic and transplantation patients. We studied the concentrations of total cholesterol, HDL-cholesterol and triglyceride in the plasma in renal (n = 48) patients. In undialyzed uraemic patients (mean serum creatinine value 439 mumol/l) the plasma cholesterol level was somewhat higher (6.47 mmol/l) than in the control (n = 27) group (5.67 mmol/l). HCL-cholesterol was lower (1.02 mmol/l vs 1.49 mmol/l) and the triglycerides higher (2.07 mmol/l vs 1.07 mmol/l) than in the control group. In dialyzed patients the cholesterol level (5.88 mmol/l) was lower but the triglyceride level higher (2.58 mmol/l) than in undialyzed patients. Renal transplantation corrected the lipid variables to a certain degree (plasma cholesterol 5.55 mmol/l, plasma HDL-cholesterol 1.27 mmol/1 plasma triglycerides 1.15 mmol/l) but HDL-cholesterol remained still somewhat lower than in the control group. No "critical level" of renal function at which the changes of lipoprotein metabolism arise, could be demonstrated, and it seems that e.g. HDL-cholesterol is low already when the diagnosis of renal insufficiency is made.


[Role of hormone receptors in the regulation of the corpus luteum]. Recent publication dealing with the hormonal regulation of the corpus luteum function has been reviewed. Hormones and receptors involved in this regulation are: LH and placental analogue HCG, prolactin and placental analogue hPL, prostaglandins, oestradiol and androgens. The hormone receptor interaction, in the luteal cells, and the role of each hormone in the modulation of the cell responsiveness has been reviewed.


[Biochemical investigations and trisomy 21 (author's transl)]. A general consideration of the pathogenesis of the various metabolic diseases which produce mental deficiency suggests that perturbation of the one carbon (folate) cycle may be important. Secondly, a review of diseases having some symptoms in common with trisomy 21 suggests the evidence of : a collagen disturbance (hypothyroidism and iminodipeptidurial) ; an oxygen disturbance (hypothyroidism and hemoglobinopathies) ; a cholinergic distrubance (Alzheimer's disease) ; a one-carbon-cycle disturbance (Lesch-Nyhan's disease). Thirdly, the peculiar pathology of trisomy 21 allows to find also a cholinergic disturbance and a disturbance close to the 10 formyl-tetrahydrololate entry of the folate cycle. Finally, an analysis of the possible effect of the excess of superoxide dismutase A and of the increase of glutathion peroxidase leads to the suspicion that a difficulty exists of dioxygenations and of non aromatic hydroxilations with a relative retardation of some FAD requiring reactions. A simplified scheme shows that these metabolic deviations could provoke a disturbance of the collagen and of synthesis of chemical mediators, in accordance with the indications furnished by the compared pathogenesis of the various affections studied. These heuristic reflexions open the way to further investigations.


Bone resorption in chronic otitis media. Bone resorption is an important aspect of chronic otitis media contributing to many complications of this disease. It is postulated that the mechanism of this localized destructive process is chemical in origin. Collagenase, lysosomal enzymes, prostaglandins, and other cell mediators are thought to induce bone resorption, but the site of action and cellular origin of these substances remains unclear. In this report, we demonstrate the location and attempt to delineate the cellular origin of two enzymes, collagenase and the lysosomal enzyme acid phosphatase in guinea pig temporal bones and human ossicles from ears containing chronic otitis media. Tissue localization of these enzymes identifies sites of active bone resorption and demonstrates the cells initiating this process. Using immunohistochemical and immunocytochemical techniques, collagenase was seen surrounding mononuclear inflammatory cells of granulation tissue at bone resorbing margins and at the periphery of osteocyte lacunae adjacent to resorbing areas. Electron microscopic data suggests that collagenase is an extracellular enzyme foun at the periphery of osteocytes. In addition, abundant acid phosphatase activity was seen in the same cells that exhibited collagenase staining, lending credence to the destructive function of these cells. The chronic inflammatory reaction found in chronic otitis media appears to activate bone destruction through the dynamic activity of mononuclear inflammatory cells and stimulates bone cells to increase their destructive biochemical functions.


gamma-Hydroxybutyric acid is not a GABA-mimetic agent in the spinal cord. gamma-Hydroxybutyric acid (GHB), a pharmacologically active central nervous system constituent, has been postulated to function as a gamma-aminobutyric acid (GABA) agonist. This hypothesis was tested directly on GABAergic synapses in isolated, superfused frog spinal cord. Addition of GHB to the superfusate produced effects on primary afferent terminals that were distinctly different from the effects of GABA. Thus, although both compounds depressed dorsal root potentials, GHB hyperpolarized terminals while GABA depolarized the same structures. The GABA responses were antagonized by bicuculline and picrotoxin, but these alkaloids did not change GHB's actions. In addition, GHB altered neither high-affinity uptake by cord slices, nor potassium-evoked release of tritiated GABA from them. GHB did not directly release GABA from spinal slices preloaded with [3H]GABA. These observations suggest that the central nervous system actions of GHB are not dependent upon its ability to activate GABAergic synapses or to modify GABAergic mechanisms.


Genetic diversity of Athabaskan Indians. If Athabaskan Indians are subdivided by linguistic group, a wide diversity in gene frequencies is disclosed. This diversity approximated that found when linguistically unrelated groups were compared. It was greater than that found for Eskimo-Aleus, even though the latter are more heterogeneous linguistically and subject to a wider variety of environmental conditions. Contiguity, geographic distance, and linguistic similarity were not reflected in similarity of gene frequencies. The gene found for Athabaskans appear to be the result of random process-survivor effects and genetic drift of small isolated groups. They appear to be of no value in detecting ethnic relationships.


[Effect of phosphorus on polymyxin B biosynthesis by B. polymyxa 1538 on media of varying makeup]. The effect of phosphorus on biosynthesis of polymyxin B by B. polymyxa 1538 was studied and its optimal concentration in the synthetic and two complex media was determined. Correlation between the culture growth and consumption of the main components, on the one hand, and concentration of phosphorus in the medium, on the other hand, was found. It was shown that the effect of phosphorus on biosynthesis of polymyxin B did not depend on the carbon source in the medium and aeration conditions.


Cytogenesis in the rat adrenal cortex: evidence for an ACTH-induced centripetal cell migration from the zona glomerulosa. The mechanism underlying cytogenesis in the adrenal cortex of ACTH-treated rats was investigated. ACTH was found to increase the number of parenchymal cells in the zona fasciculata as well as that of "S" phase cells in the inner half of the zona glomerulosa and in the outer third of the zona fasciculata. The fate of 3H-thymidine labelled cells in ACTH-administered rats was also autoradiographically followed. The results are consistent with the hypothesis that the ACTH-provoked hyperplasia in the zona fasciculata involves the centripetal migration of newly-formed parenchymal cells from the zona glomerulosa.


The serum protein pattern in primary hepatoma and amoebic liver abscess. A study of the serum proteins pattern of 30 patients with primary liver cell carcinoma and 11 with amoebic liver abscess was carried out. When compared with controls significant differences were found for both conditions in the values of pre-albumin, transferrin, albumin, haptoglobin, alpha 2-macroglobulin, and alpha 2HS-glycoprotein. In the differential diagnosis of amoebic liver abscess and primary hepatic carcinoma, the estimation of albumin, alpha 1-acid glycoprotein, haptoglobin ceruloplasmin, alpha 2H2-glycoprotein and transferrin was found helpful.


[Coproantibodies in intestinal amebiasis]. E. histolytica tissue invasion induces IgG and IgM and probably IgA serum antibody formation of different types in the intestinal mucosa. 21 children with intestinal invasive amebiasis were studied. Samples were taken from peripheral blood and stools during diagnostic studies and three weeks later. Contraimmunoelectrophoretic antibody determinations and indirect hemagglutination tests IH were made in an attempt to demonstrate its true nature. During the acute phase (first sample), results showed antibodies against amoeba in stools by CIE in 57 per cent and by IH in 80 per cent; in serum, results were 38 and 76 per cent respectively. In the second sample and three weeks after diagnosis was established a decrease in the number of patients with coproantibodies against amoeba was observed and the number of patients with serum antibodies increased. We are also providing evidence that supports the IgG and IgA nature of coproantibodies against amoeba. These results suggest the presence of a local immunological reaction in invasive amebiasis.


[Acute infectious gastroenteritis. Etiology and its correlation with clinical manifestations and fecal mucus]. 343 children with acute diarrhea were studied from january 1976 to september 1977. Rotavirus was the agent most frequently isolated (18 per cent) followed by Shigella (12 per cent) and Salmonella (10 per cent). Enterotoxigenic E. coli was identified in 8 per cent and invasive E. coli only in two cases. 80 per cent of isolated rotavirus fell in the neonate group and 25 per cent in the infant group. Diarrhea caused by rotavirus had a short duration, fever was negligible and abundant liquid stools were present without leukocytes in the fecal mucus and with a high percentage (48 per cent) of transient lactose intolerance. Polymorphonuclear leukocytes were found in fecal mucus in 75 per cent of cases caused by Shigella and only in 40 per cent of cases where the causal agent was Salmonella.


Lipids and lipoproteins in subjects at 1,000 and 3,500 meter altitudes. To assess the relationship between altitude, atherogenic, and anti-atherogenic lipoprotein cholesterols (low- [C-LDL] and high [C-HDL] density lipoprotein cholesterols, respectively), 136 and 94 Venezuelan Mestizos living at 1,000 and 3,500 in elevation were studied. The two groups did not differ in regard to height, weight, ethnic origin, social or economic status, nutritional patterns, age, or occupation. Both groups had a high level of daily physical exertion, an imperative in their subsistence rural agricultural economy. Due to the mountainous terrain, high altitude residents were thought to have increased levels of physical activity. Males and females at high altitude had significantly lower plasma total cholesterol and C-LDL levels, and slightly lower C-HDL levels than those at low altitudes. It is speculated that reduced coronary heart disease event rates at high altitude might be related to lower levels of the atherogenic lipoprotein cholesterol, C-LDL.


Effects of silica dust inhalation on the susceptibility of mice to influenza infection. Mice were exposed to silica dust for durations of up to 36 wk. At intervals of 15, 21, 27, 33, and 36 wk, the ability of the splenic lymphocytes to respond to T and B cell mitogens were determined, and their ability to resist influenza virus infections was determined after 3 and 20 wk of exposure. These exposures had a depressing effect on the response to T cell mitogens at 21 and 27 wk of silica exposure, but there was no effect on the T cell responses after 15, 33, and 36 wk or on B cell responses after all exposure times. No changes could be detected in the ability of the mice to resist pulmonary influenza virus infections or in the survivors ability to form HI antibodies against this virus.


Fluphenazine and social therapy in the aftercare of schizophrenic patients. Relapse analyses of a two-year controlled study of fluphenazine decanoate and fluphenazine hydrochloride. The ability of long-acting fluphenazine decanoate and oral fluphenazine hydrochloride to forestall relapse among newly discharge schizophrenic patients is examined in the context of high and low degrees of social therapy (ST). A total of 105 patients were randomly assigned to the various treatments and maintained under controlled conditions for two years or until relapse. Relapse rates for all treatments remained traditionally high. Relpase rates for long-acting fluphenazine decanoate and oral fluphenazine hydrochloride are nearly identical in the first year, indicating that drug noncompliance does not adequately explain early schizophrenic relapse. However, patients who received long-acting fluphenazine decanoate and ST have a reduced risk of relapse over time. Relapsers who received long-acting fluphenazine decanoate appeared more affectively disturbed than other relapsers, yet both groups were diagnostically and symptomatically equivalent prior to treatment. Personal discomfort and intrafamilial stress are important predictors.


[Testing of solid tumors in childhood for sensitivity against cytostatic agents using an autoradiographic in vitro method (short-term method) (author's transl)]. The sensitivity of 37 solid tumours of children was tested in vitro towards cytostatic agents by means of an autoradiographic short-term method. Sensitivity was measured as the magnitude of inhibition of 3 H-thymidine incorporation. The test was performed with the cytotoxic agents Cyclophosphamide, Trenimon, Bleomycin, Adriamycin, Daunomycin, Actinomycin D, and Cytosin-Arabinosid in 9 Wilms' tumours, 9 neuroblastomas, 7 non-Hodgkin-lymphomas, 5 osteogenic sarcomas, 3 soft tissue sarcomas, and 4 special tumours. None of the tumours is resistant to all cytotoxic substances. The tumours show a marked individual sensitivity pattern, and, with few exceptions, they are sensitive against 2 or more cytostatics. This behaviour is explained mainly by the usually high proliferative activity of dysontogenetic tumours, malignant lymphomas and various sarcomas. The possibilities and limits of the short-term methods for sensitivity-testing are discussed critically and in detail. For the evaluation of the results of in vitro testing and of in vivo effectiveness the close coreelations are not always taken into consideration between the type of cytostatic agent and effect on tumour metabolism, cytostatic agent and proliferation kinetics of the tumour as well as the effect of the cytostatics and the nucleic acid precursor used for the test. Despite the methodological limitations preclinical testing should be preferred in comparison with unselected chemotherapy.


Total "internal" hemipelvectomy.

Total "internal" hemipelvectomy. Several authors have described resections of tumorbearing parts of the pelvis including reconstructive plastic surgery. With extensive tumor growth present mutilating hemipelvectomies often have been the last resort. Even the most sophisticated prostheses hardly ever enabled the patients to walk again. We have therefore performed total "internal" hemipelvectomies in three cases. The necessary substitute consisted of half a pelvis made of polyacetal resin produced from a given model. The model's measures and shape was established by computer tomograms thus resulting in an individually shaped pelvis half. The operative procedure aims at the maintenance of osseous muscular insertions and origins and their reconstruction. Utmost care has to be taken in avoiding damage to nerves and vessels. Fixation of the implant is performed by screws aiming through the sacroiliac joint into ala and corpus of the sacrum and corticocancellous grafts incorporated in the implant. The symphyses are united by means of plate fixation. The hip joint is substituted by a total prosthesis. With the procedure described we have managed three cases of extensive malignant tumors of the pelvis. One patient suffered a nerve lesion and skin necrosis with infection. The two remaining patients are capable of weight-bearing and standing on the inflicted leg alone.


Studies on the attachment of herpes simplex virus. Effect of trypsin treatment. The adsorption of herpes simplex virus (HSV) to monolayers of trypsinized and mock-treated GMK-AH1 cells was studied by means of a microplaque technique. Plaque formation was nearly abolished for c:a 20 minutes and full sensitivity regained at 6--8 hours post trypsinization. The return of sensitivity to HSV was inhibited by cycloheximide and incubation at 4 degrees C. The sum of PFU in the supernatant and PFU successe modifying agents were without effect on the adsorption of HSV.


[Etiology of combined inhalational hydrocyanic acid and carbon monoxide poisoning]. A young man was found dead in a kitchen, that was partly burnt. Autopsy revealed, as cause of death, a combined intoxication following inhalation of carbon monoxide and hydrocyanic acid. Own investigations on the pyrolysis of pieces of furniture found in the kitchen (plastic plates containing melamine and plates containing formaldehyde) showed, that hydrocyanic acid was liberated through combustion of such substances and inhaled by the victim. The poisoning picture is discussed, and discussion includes especially considerations on the peculiar sensitivity of the brain toward the action of hydrocyanic acid and the relative insensitivity of the heart muscle. It is thought that the cause of such sensitivity difference lies in the physiological differences of the intracellular energy production. Finally the dangers of combustion gases developing from burning plastic materials are reemphasized.


Taxonomic studies on strains of avian infectious bronchitis virus using neutralisation tests in tracheal organ cultures. The antigenic relationships of 24 strains of avian infectious bronchitis virus (IBV) were investigated by serum neutralisation tests performed in chick embryo tracheal organ cultures. The serum dilution that neutralised 100 median ciliostatic doses (CD50) of virus was estimated from the linear relationship between varying concentrations of each virus strain and the neutralisation titre of homologous antiserum; this dilution defined 1 antibody unit. Antisera diluted to contain 20 antibody units were then tested by neutralisation against 1.5--2.5 log10 CD50 of each strain. Clusters of both strains and antisera in turn were established by methods of numerical taxonomy using as measures of resemblance Euclidean distance and correlation coefficient, and by analysis by principal components. These analyses identified a group of 8 similar strains; neutralisation of the remaining 16 strains was slight. Similar results were obtained by classifying antisera, except that a further group of 3 antisera was demonstrated, each having a neutralising capacity for most strains. Implications for vaccine formulation are discussed.


The effect of pyrophosphate analogues on influenza virus RNA polymerase and influenza virus multiplication. Analogues of pyrophosphate have been tested as inhibitors of influenza virus-RNA polymerase activity in cell-free assays. The most active compound, phosphonoformic acid (PFA), reduced the polymerase activity to 50 per cent at a concentration of 20 muM. The inhibition was dependent on the type of divalent cation present in the assay. PFA at a concentration of 400 muM also inhibited the influenza virus plaque formation by 90 per cent.


A new calicivirus isolated from a marine mammal. A new serotype of calicivirus, designated as San Miguel sea lion virus type 6 (SMSV-6), was isolated from vesicular lesions on the flipper of a California sea lion pup. Serologic studies show that SMSV-6 neutralizing antibodies (SN) occur frequently among California sea lions and occasionally among northern fur seals. Feral swine, 1- to 6-week elephant seal pups and grey whales tested negative for SMSV-6 antibody.


[Radioautographic study of the localization of 3H-estradiol, 3H-histamine, and 3H-cyclic adenosine monophosphate in rat uterus]. The method of radioautography has demonstrated that 3H-estradiol adheres to the nuclei of some myometrium cells, 3H-histamine is accepted by the cytoplasm of the majority of the myometrium cells, 3H-cyclic-AMP is selectively bound by the endothelial cells of capillaries and small vessels in all layers of the uterus. From the data obtained it is possible to conclude that estradiol and its mediators, histamine and cyclic-AMP, specifically interact with different cells and with different cell structures.


[Intrauterine infections in the perinatal period]. The results of examinations of autopsy materials in Leningrad and the data from the literature indicate that infectious diseases occupy a significant place in perinatal mortality. It is emphasized that if the autopsy materials are not sufficiently examined using not only light but also immunofluorescent microscopy, a considerable portion of these infections is not diagnosed and the pathological proprocess in such cases in considered to be aspiration pneumonia, hyaline membrane disease, etc. The paper briefly characterizes intrauterine bacterial infections (listeriasis, syphilis, staphylococcal infections, etc.) with special reference to virus diseases (cytomegaly, herpes infection, lesions caused by respiratory viruses, etc) and mycoplasmosis. It is shown that a relatively high incidence of lesions is caused by respiratory viruses and particularly mycoplasma. The possibility of protozoan and mycotic lesions is also indicated. The frequent occurrence of combined intrauterine infections is emphasized.


Prevalence of antibody to hepatitis A virus in Sri Lanka. Antibody to hepatitis A virus was studied by immune adherence hemagglutination using 287 serum specimens collected in 1975 by random sampling from healthy individuals living in Colomobo, Sri Lanka. The overall prevalence of antibody to the virus was 76.9% and the prevalence between males and females was approximately the same. The age-specific prevalence of the antibody indicated that hepatitis A in the area is mainly an infection during the early childhood.


Immediate postembolization excision of glomus jugulare tumors: advantages of new combined techniques. Preoperative percutaneous transfemoral catheter embolization of feeding vessels in glomus jugulare tumors, followed by immediate application of standard surgical techniques, presents the treatment of choice, allowing meticulous microsurgery with virtually complete hemostasis. Therefore, the surgeon can operate in a bloodless environment throughout the compressed and intricate anatomic field, amidst such important yet vulnerable structures as cranial nerves, inner ear, carotid artery, jugular bulb, venous sinuses, and dura, while reducing surgical error and functional deficit for the patient. Review of the last 11 cases of glomus jugulare tumors at UCLA shows that even extensive Alford grade 2 tumors of the middle ear, jugular bulb, and mastoid had only minor blood losses with this combined technique of embolization-immediate surgery, as compared with earlier surgical methods. Pertinent literature on glomus jugulare and its treatment is reviewed. Combined embolization and immediate surgery offer the best approach for treatment of resectable glomus jugulare tumors.


Arteriolar rarefaction in hypertension.

Arteriolar rarefaction in hypertension. The functional rarefaction of small arterioles previously reported at 6 weeks of age in the SHR has been shown to be a structural decrease in arteriolar density as well. In addition, the high "occlusive tone" normally found in young normotensive rats is observed in the older SHR. This elevated tone contributes to a functional rarefaction in the older SHR arterioles. The mechanism for the rarefaction appears to be related to the cAMP/cGMP second messenger system.


Transfer of carbohydrates on to nascent glycoprotein of vesicular stomatitis virus. I studied the glycosylation in vivo of a viral envelope protein, the glycoprotein of vesicular stomatitis virus (VSV), by pulse labelling of virus-infected HeLa cells with 3H-labelled monosaccharides (mannose, glucosamine). Radioactivity was incorporated into the fraction of membrane-bound polyribosomes, although metabolic conversion of [3H]-mannose into amino acids was negligible. Dissociation of bound polyribosomes revealed that the radioactively co-purified with the peptidyl-tRNA. The nascent peptides were released by alkaline hydrolysis, immunoprecipitated and analysed by polyacrylamide-gel electrophoresis. It is apparent from the size distribution of the [3H]mannose-labelled nascent chains that attachment of carbohydrate starts when approximately half of the amino acid sequence of the viral glycoprotein has been synthesized.


Interaction of phosphorylase b with eosin. Influence of substrate and effectors on eosin-enzyme complexes. The interactions of rabbit muscle glycogen phosphorylase b with Eosin (2',4',5',7'-tetrabromofluorescein) was studied. Eosin was found to be an effective inhibitor of the enzyme. The inhibition constants for the dye were estimated to be approx. 36 and 60 microM with respect to AMP and glucose 1-phosphate respectively. The binding of Eosin to phosphorylase b is accompanied by a red-shift of about 12 nm in the dye absorption-spectrum maximum, indicating low-polarity binding sites on the enzyme molecule for the dye. The absorbance in the difference absorption maximum at 537 nm was utilized to follow the conjugation of phosphorylase b with Eosin. Scatchard plots of the titration data revealed the existence of at least two classes of binding sites on the protein molecule for Eosin, and the dissociation constants measured in Tris/HCl buffer, pH 7.0 (IO.091), were 7.7 and 41.7 microM respectively. The influence of the substrates and effectors on Eosin-enzymes complexes was used to study the ligand-phosphorylase b interactions. IMP displaced the dye completely from the enzyme, indicating that there are two IMP-binding sites per phosphorylase b monomer. AMP binding to the enzyme with respect to Eosin concentration is of two types: a non-competitive one for the high-affinity site for AMP and a competitive one for the low-affinity site for the activator. The effects of glucose 6-phosphate, ATP, Pi and glycerol 2-phosphate in the system are in according dance with a partially competitive model. Glucoes 1-phosphate and UDP-glucose appear to affect only the high-affinity site for Eosin, whereas glucose and glycogen have no effect on Eosin-phosphorylase b complexes. Our results suggest that Eosin can be used as an efficient optical probe for studying the phosphorylase b system.


Separation and purification of the alkaline phosphatase and a phosphodiesterase from Halobacterium cutirubrum. 1. Halobacterium cutirubrum alkaline phosphatase is associated in crude extracts with a phosphodiesterase. 2. The enzymes were stabilized in buffers containing both (NH4)2SO4 and 10 mM-Mn2+. 3. Adsorption chromatography on Sepharose 6B/agarose-gel columns in the presence of 1.4M-(NH4)2SO4 gave a phosphatase-free phosphodiesterase and the alkaline phosphatase associated with some phosphodiesterase activity. 4. Further chromatography of the separated enzymes gave a good recovery of greater than 600-fold purified phosphodiesterase and greater than 3000-fold purified alkaline phosphatase. 5. The requirements of these enzymes and their relationship to each other was examined. 6. A detailed study showed that the alkaline phosphatase was adsorbed at least partially to agarose and dextran columns at all (NH4)2SO4 concentrations from 0.25 to 2M. 7. In contrast, no adsorption of the enzyme or protein standards was evident in 2.5M-KCl/l M-NaCl or 0.25 M-KCl/0.1 M-NaCl, in agreement with previous studies by Louis, Peterkin & Fitt [(1971) Biochem. J. 121, 635-641], thus confirming the validity of gel filtration in 2.5 M-KCl/1 M-NaCl as a method for determining the approximate molecular weights of extremehalophile proteins.


The development of SS'-polymethylenebis(methanethiosulphonates) as reversible cross-linking reagents for thiol groups and their use to form stable catalytically active cross-linked dimers within glyceraldehyde 3-phosphate dehydrogenase. The synthesis of a series of SS'-polymethylenebis(methanethiosulphonates) including the pentane, hexane, octane, decane and dodecane derivatives is described. These derivatives were synthesized by condensation between dibromoalkanes and potassium methanethiosulphonate in refluxing methanol and this seems an especially versatile reaction for the synthesis of asymmetric thiosulphonate derivatives. The synthesis of SS'-[1,8-3H4]-octamethylenebis(methanethiosulphonate) was also perfomed. Cross-linking was demonstrated in the four enzymes lactate dehydrogenase, phosphofructokinase, pyruvate kinase and glyceraldehyde 3-phosphate dehydrogenase. For all four enzymes cross-linking was efficiently reversed by reducing conditions in denaturing solvents. The reaction with glyceraldehyde 3-phosphate dehydrogenase was unique in that only the cross-linked dimer was produced in significant amounts (greater than 90% of total products as dimer). This reaction was followed in detail with radioactive cross-linking reagent. Inhibition of enzyme activity was extremely fast and showed an asymmetric distribution of enzyme activity on subunits. Thus complete modification of only one subunit resulted in up to 75% inhibition of enzyme activity. Reaction of glyceraldehyde 3-phosphate dehydrogenase with 1.25 mol of SS'-octamethylenebis(methanethiosulphonate) per mol of enzyme subunit produced two species of protein. The first species was obtained in 20% yield and was only partially re-activated on mild reduction with 2-mercaptoethanol. The second species was isolated in 66% yield and was completely re-activated on mild reduction. Before reduction there was 4 mol of inhibitor per tetramer for the latter species, and more than 95% of the enzyme was present as a dimer on non-reducing electrophoresis. After mild reduction 2 mol of inhibitor was still bound per tetramer, the enzyme was now catalytically active and the dimer was still the major structure on non-reducing electrophoresis. Thus mild reduction of SS'-octamethylenebis(methanethiosulphonate-treated glyceraldehyde 3-phosphate dehydrogenase enabled the production of active enzyme in which there is a stable cross-link across one of the molecular axes of the tetrameric enzyme. This cross-link was only reversed if reduction was performed when the enzyme was denatured. The molecular weight of cross-linked and re-activated cross-linked glyceraldehyde 3-phosphate dehydrogenase was established as 144000 (tetramer) by sucrose-density-gradient centrifugation. These observations are interpreted in terms of the molecular structure of glyceraldehyde 3-phosphate dehydrogenase.


Detergent activation and solubilization of 2':3'-cyclic nucleotide 3'-phosphodiesterase from isolated myelin and c6 cells. Several detergents were investigated for their ability to increase activity of 2':3'-cyclic nucleotide 3'-phosphodiesterase in isolated myelin. The ability of Triton X-100 and Sulfobetaine DLH to solubilize the enzyme was also examined. Solubilization with Triton X-100 was only effective in the presence of salt, for example with NaCl 51% of the activity was solubilized. A single extraction with Sulfobetaine DLH yielded slightly more solubilized enzyme and did not require added salt. Both activation and solubilization of 2':3'-cyclic nucleotide 3'-phosphodiesterase appeared to be similarly dependent on detergent concentration, suggesting a common action of the detergent in the two processes. Myelin basic protein was solubilized more readily than the enzyme. In contrast with the enzyme in myelin, 2':3'-cyclic nucleotide 3'-phosphodiesterase activity in C6 cells was not increased in the presence of Triton X-100, and was partially solubilized by either Triton X-100 or NaCl alone. No myelin basic protein could be detected in C6 cells by radioimmunoassay.


Neuraminidase inhibition by chemically sulphated glycopeptides. Chemically sulphated glycopeptides (derived from pig duodenal mucosa) inhibited Clostridium perfringens neuraminidase (EC 3.2.1.18) activity in a pH-dependent manner. Analysis of inhibition kinetics data indicated that, although the enzyme inhibition could not be categorized into any of the classical types of inhibition, it could be interpreted as a function of the size and shape of the substrates used. The enzyme activity was inhibited by 86% and 40% when tested with bovine submaxillary-gland mucin (mol. wt. 4 x 10(5)-40 x 10(5) and N-acetylneuraminyl-lactose (mol. wt. 633) as substrates respectively. Presence of sulphated glycopeptide did not affect the binding of N-acetylneuraminic acid (mol. wt. 309), a competitive inhibitor of Vibrio cholerae neuraminidase, to the enzyme active site. The enzyme inhibition was thus considered to be due to steric hindrance as a consequence of the non-specific interactions between the enzyme molecule and polyanionic sulphated glycopeptide affecting the differential accessibility of the substrate molecules to the enzyme active site. The enzyme-inhibitor interaction could be suppressed by rapid and many-fold dilution of the reaction mixture, by concurrent addition of the inactive enzyme or by partial removal of the sulphate esters from the sulphated glycopeptide molecule by the action of Helix pomatia arylsulphatase (EC 3.1.6.1).


Effect of O-sulphate groups in lactose and N-acetylneuraminyl-lactose on their enzymic hydrolysis. 1. Lactose 6'-O-sulphate, N-acetylneuraminyl-(alpha 2 leads to 3)-D-lactose 6'-O-sulphate, N-acetylneuraminyl ?-O-sulphate-(alpha 2 leads to 3)-D-lactose 6'0-O-sulphate, N-acetylneuraminyl ?-O-sulphate-(alpha 2 leads to 6)-D-lactose and N-acetylneuraminyl-(alpha 2 leads to 3)- and -(alpha 2 leads to 6))-lactose 6'-O-sulphate were prepared by chemical sulphation of lactose, N-acetylneuraminyl-lactose and tis isomers by using pyridine-SO3 reagent. 2. Significant kinetic differences were observed in the enzymic hydrolysis of the sulphated derivatives compared with unsubstituted substrates. 3. In the case of reactions catalysed by rat liver lysosomal and Clostridium perfringens neuraminidases (EC 3.2.1.18), the presence of an O-sulphate group in the N-acetylneuraminyl moiety affected the reaction by decreasing the Km and the Vmax, its presence in the galactosyl moiety affected the reaction by decreasing the Km and increasing the Vmax. and its presence in both N-acetylneuraminyl and galactosyl moieties decreased the Km and the Vmax. of the reaction. 4. Mixed-substrate reaction kinetic data indicated competition between the sulphated and unsubstituted substrates for the same active sites on the neuraminidase molecule. 5. Lactose 6'-O-sulphate neither behaved as a substrate nor acted as an inhibitor with respect to unsubstituted lactose and p-nitrophenyl beta-D-galactopyranoside when tested with lactase of suckling rat intestine and Escherichia coli beta-D-galactosidase (EC 3.2.1.23). 6. Preliminary investigation also indicated that, whereas glucose 6-O-sulphate and glucose 3-O-sulphate were were neither substrate nor inhibitor of glucose oxidase (EC 1.1.3.4), galactose 6-O-sulphate was oxidized half as fast as unsubstituted galactose by galactose dehydrogenase (EC 1.1.1.48).


Regulation of kinase reactions in pig heart pyruvate dehydrogenase complex. 1. Pig heart pyruvate dehydrogenase complex is inactivated by phosphorylation (MgATP2-) of an alpha-chain of the decarboxylase component. Three serine residues may be phosphorylated, one of which (site 1) is the major inactivating site. 2. The relative rates of phosphorylation are site 1 greater than 2 greater than site 3. 3. The kinetics of the inactivating phosphorylation were investigated by measuring inactivation of the complex with MgATP2-. The apparent Km for the Mg complex of ATP was 25.5 microM; ADP was a competitive inhibitor (Ki 69.8 microM) and sodium pyruvate an uncompetitive inhibitor (Ki 2.8 microM). Inactivation was accelerated by increasing concentration ratios of NADH/NAD+ and of acetyl-CoA/CoA. 4. The kinetics of additional phosphorylations (predominantly site 2 under these conditions) were investigated by measurement of 32P incorporation into non-radioactive pyruvate dehydrogenase phosphate containing 3-6% of active complex, and assumed from parrallel experiments with 32P labelling to contain 91% of protein-bound phosphate in site 1 and 9% in site 2. 5. The apparent Km for the Mg complex of ATP was 10.1 microM; ADP was a competitive inhibitor (Ki 31.5 microM) and sodium pyruvate an uncompetitive inhibitor (Ki 1.1 mM). 6. Incorporation was accelerated by increasing concentration ratios of NADH/NAD+ and of acetyl-CoA/CoA, although it was less marked at the highest ratios.


1H nuclear-magnetic-resonance spectra of the methyl group of the acetamido moiety and the structure of acid glycosaminoglycans in solution. The 1H resonances of the methyl group in the acetamido moiety of several types of glycosaminoglycans are reported at 300 MHz in 2H2O. Dermatan sulphates with various L-iduronate/D-glucuronate ratios are compared with chrondroitin sulphates with various contents and positions of substitution of sulphate esters. Hyaluronate oligomers are compared with 2-acetamido-2-deoxy-D-glucose, and with heparan sulphate and keratan sulphate. The major determinant of the chemical shift of the acetamido methyl resonance is the closeness of approach between carboxy groups and the acetamido group, in agreement with chemical evidence based on periodate-oxidation kinetics.


Metabolism and biological activity of 25-fluorocholecalciferol, 24-dehydrocholecalciferol and 25-dehydrocholecalciferol in the rat. Three side-chain analogues of cholecalciferol (vitamin D3) modified at C-25, namely 25-fluorocholecalciferol, 24-dehydrocholecalciferol and 25-dehydrocholecalciferol, conceived as potential inhibitors of the cholecalciferol 25-hydroxylase have been prepared and tested in the rat. These compounds markedly diminish conversion in vivo of cholecalciferol into 25-hydroxycholecalciferol, but are not antagonists of vitamin D action, because they themselves possess significant biological activity in vivo. Each compound is capable of stimulating the intestinal transport of calcium and the mobilization of calcium from bone in vitamin D-deficient rats. Biological responses equivalent to those generated by a physiological dose of cholecalciferol (0.05 microgram) are produced, however, only when the analogues are administered at high doses (5-50 microgram). The biological activity of 24-dehydrocholecalciferol and 25-dehydrocholecalciferol is shown to result from conversion, in vivo, to the natural hormone, 1 alpha,25-dihydroxycholecalciferol, whereas 25-fluorocholecalciferol is metabolically activated in the rat by hydroxylation to 1 alpha-hydroxy-25-fluorocholecalciferol. This latter conversion is the first reported example of the 1 alpha-hydroxylation of a vitamin D compound lacking the 25-hydroxy group.


Determination of vitamin D and its metabolites in plasma from normal and anephric man. A multiple assay capable of reliably determining vitamins D(2) and D(3) (ergocalciferol and cholecalciferol), 25(OH)D(2) (25-hydroxyvitamin D(2)) and 25(OH)D(3) (25-hydroxyvitamin D(3)), 24,25(OH)(2)D (24,25-dihydroxyvitamin D), 25,26(OH)(2)D (25,26-dihydroxyvitamin D) and 1,25(OH)(2)D (1,25-dihydroxyvitamin D) in a single 3-5ml sample of human plasma was developed. The procedure involves methanol/methylene chloride extraction of plasma lipids followed by separation of the metabolites and purification from interfering contaminants by batch elution chromatography on Sephadex LH-20 and Lipidex 5000 and by h.p.l.c. (high-pressure liquid chromatography). Vitamins D(2) and D(3) and 25(OH)D(2) and 25(OH)D(3) are quantified by h.p.l.c. by using u.v. detection, comparing their peak heights with those of standards. 24,25(OH)(2)D and 25,26(OH)(2)D are measured by competitive protein-binding assay with diluted plasma from vitamin D-deficient rats. 1,25(OH)(2)D is measured by competitive protein-binding assay with diluted cytosol from vitamin D-deficient chick intestine. Values in normal human plasma samples taken in February are: vitamin D 3.5+/-2.5ng/ml; 25(OH)D 31.6+/-9.3ng/ml; 24,25(OH)(2)D 3.5+/-1.4ng/ml; 25,26(OH)(2)D 0.7+/-0.5ng/ml; 1,25(OH)(2)D 31+/-9pg/ml (means+/-s.d.). Values in two normal human plasma samples taken in February after 1 week of high sun exposure are: vitamin D 27.1+/-7.9ng/ml; 25(OH)D 56.8+/-4.2ng/ml; 24,25(OH)(2)D 4.3+/-1.6ng/ml; 25,26(OH)(2)D 0.5+/-0.2ng/ml. Values in anephric-human plasma are: vitamin D 2.7+/-0.8ng/ml; 25(OH)D 36.4+/-16.5ng/ml; 24,25(OH)(2)D 1.9+/-1.3ng/ml; 25,26(OH)(2)D 0.6+/-0.3ng/ml; 1,25(OH)(2)D was undetectable.


Effect of growth conditions on the involvement of cytochrome c in electron transport, proton translocation and ATP synthesis in the facultative methylotroph Pseudomonas AM1. The stoicheiometry of proton translocation, the amounts of cytochromes firmly bound to membranes, and cell yields with respect to succinate and O(2) have been measured in the facultative methylotroph Pseudomonas AM1 and in the mutant lacking cytochrome c (mutant PCT76) during carbon-limited growth and carbon-excess growth. -->H(+)/O ratios during endogenous respiration of about 4 were measured in wild-type bacteria grown in carbon-excess conditions, and in the mutant in all growth conditions. During methanol- or succinate-limited growth of wild-type bacteria the -->H(+)/O ratio increased to about 6. Cell yields with respect to succinate and O(2) were higher in wild-type than in the mutant lacking cytochrome c by an amount suggesting loss in the mutant of 30% of the ATP-generating capacity of wild-type bacteria. During carbon-limited growth on methanol or succinate some cytochrome c was tightly bound to bacterial membranes, whereas none was tightly bound in bacteria grown in batch-culture or in NH(4) (+)-limited conditions. It is proposed that the role of cytochrome c in Pseudomonas AM1 depends on growth conditions and hence on the ;needs' of the bacteria. During growth in carbon-excess conditions it is only required for methanol oxidation, mediating between methanol dehydrogenase and cytochrome a/a(3). In these conditions oxidation of NADH and succinate by way of cytochrome b and cytochrome a/a(3) occurs without the mediation of cytochrome c. This is the only route for oxidation of NADH and succinate in the cytochrome c-deficient mutant in all growth conditions. During carbon-limited growth the cytochrome c becomes bound to the membrane in such a way that it can mediate between cytochromes b and a/a(3), hence becoming involved in proton translocation and ATP synthesis during NADH and succinate oxidation. An alternative possibility is that in wild-type bacteria the cytochrome c is always involved in electron transport, but that its involvement in measurable proton translocation only occurs in carbon-limited conditions.


Translation of embryonic-chick tendon procollagen messenger ribonucleic acid in two cell-free protein-synthesizing systems. Embryonic-chick tendon poly(A)-containing RNA was translated in the wheat-germ and mRNA-dependent rabbit reticulocyte-lysate systems. The ability of each system to synthesize polypeptides similar to pro-alpha chains of collagen was tested on the bases of electrophoretic mobility and susceptibility to highly purified bacterial collagenase. Very small amounts of polypeptides in the size range of pro-alpha chains were synthesized in the wheat-germ system, whereas efficient synthesis of two polypeptides similar to pro-alpha1 and pro-alpha2 chains was achieved in the reticulocyte lysate. The collagenous nature of the major high-molecular-weight products synthesized was demonstrated by their susceptibility to collagenase and ability to act as a substrate for purified collagen proline hydroxylase. Determinations of the relative amounts of these translation products suggest that the 2:1 ratio of pro-alpha1 and pro-alpha2 chains found in type I procollagen is reflected in proportional amounts of translatable mRNA for pro-alpha1 and pro-alpha2 chains. Comparisons of the electrophoretic mobilities of hydroxylated and unhydroxylated reticulocyte-lysate translation products were made with appropriate standards of hydroxylated and unhydroxylated procollagen polypeptides. The results suggest that, in common with a number of secreted proteins, procollagen is synthesized as pre-pro molecules consistent with the ;Signal Hypothesis'.


Ultrastructural examination of blood dialysate effect in experimental carbon monoxide poisoning of the heart muscle. The effects of acute carbon monoxide poisoning on the ultrastructure of the muscle had been investigated. Dogs were exposed to 10.1 vol.% carbon monoxide by intracoronary perfusion for 10, 20 and 30 min. The ultrastructural changes and cytochrome-oxidase activity reduction were evident after 10 min and were most prominent after 20 and 30 min. Degenerative changes of mitochondria, pericapillary and interfibrillar edema, sarcoplasmic reticulum dilatation, glycogen content decrease and in local areas a total or partial necrosis of myofibrils could characteristically be observed. The cytochrome-oxidase and glycose-6-phosphate dehydrogenase showed a significant decrease. A dialysis concentrate from deproteinised calf blood (Solcoseryl, Actihaemyl) proved to inhibit morphological and enzymatical pathological changes; the destruction was slowed and prevented. On this basis the protective effect of Solcoseryl may be of considerable importance.


Stimulatory effect of ginsenosides on DNA, protein and lipid synthesis in rat bone marrow and participation of cyclic nucleotides. Effects of several kinds of ginsenosides, saponins from Panax ginseng on DNA, RNA, protein and lipid synthesis in bone marrow were investigated. Single i.p. injection of 0.5--1 mg/100 g body weight of ginsenosides Rb2, Rc, Re and Rg1 4 h prior to the sacrifice increased DNA synthesis in bone marrow cells. RNA, protein and lipid synthesis were also increased. The direct addition of ginsenosides Rb1, Rb2 and Rc mixture (GNS) enhanced DNA synthesis. Cyclic AMP levels in bone marrow cells were decreased 20 min after i.p. injection of ginsenosides Rb2, Rc and Rg1 and the direct addition of ginsenosides Rb2, Rc and Rg1 also decreased cyclic AMP levels. While cyclic GMP levels were increased by administration of ginsenosides Rb2, Re and Rg1. Relationship between chemical structure and actions of ginsenosides and the role of cyclic nucleotides in the stimulatory action of ginsenosides on DNA synthesis in bone marrow cells were discussed.


Further studies on the antipyretic action of polymyxin B in pyrogen-induced fever. A study of the antipyretic effect of polymyxin B was undertaken to determine how this agent reduces fever in rabbits. It involved the effects of the drug: (1) on fever induced by exogenous pyrogenes (E. coli lipopolysaccharide, synthetic double-stranded ribonucleic acid, sodium nucleinate from yeast) and leucocytic pyrogen, (2) on the release of endogenous pyrogen in vivo and in vitro, and (3) on leucocytic and exogenous pyrogens in vitro. The results indicate that polymyxin B produces an antipyretic effect in endotoxin-induced fever primarily by an interaction of this cationic macromolecule with the anionic endotoxin molecule. Further it is likely that polymyxin B inhibits endogenous pyrogen synthesis and/or release from polymorphonuclear leucocytes.


[Effect of halofenate on triglyceride and uric acid levels, coagulation and platelet behaviour in patients with hyperlipoproteinemia type IV and hyperuricemia (author's transl)]. From experimental work, an influence of a drug with hypolipidemic and hypouricemic acition on blood coagulability and platelet function may be expected. Consequently, if these effects were demonstrable in man the drug could be assumed to reduce the tendency to develop thrombosis and atherosclerosis in patients with hyperlipidemia and hyperuricemia. In the study reported, the effect of 2-acetamidoethyl-(p-chlorophenyl)-(m-trifluoro-methylphenoxy)-acetate (halofenate) was investigated in 14 patients suffering from hyperlipoproteinemia type IV and hyperuricemia. Platelet aggregation and adhesiveness, plasma levels of triglycerides, cholesterol, uric acid, and clotting factors were regularly examined during a three-month double blind trial. While uric acid and triglyceride levels decreased, no influence of the drug treatment could be observed on platelet function and blood coagulability by the laboratory methods used.


Polymyxin B-induced oedema in hind paw of the rat as an assay for antiinflammatory drugs. A method is presented for measuring the volume of the oedema induced by intraplantar infection of polymyxin B, a polyenic antibiotic, in the hind paw of the rat. Peak values of the oedema reached the maximum within 30 min to 1 h after injecting the drug and decreased with time until the 24th h, when it had almost disappeared. The phlogogenic effect of polymyxin B seems to be related to that induced by serotonin and histamine, rather than to the reaction provoked by carrageenin or prostaglandins. The oedema was inhibited by pretreating the animals p.o. with both steroidal and non-steroidal antiinflammatory agents. The inhibitory effect of antiinflammatory drugs was found to be dose-dependent.


Activity of tolmetin on levels of cyclic nucleotides in experimental pleurisy. Experimental pleurisy induced in rats with an intrapleural injection of 0.2 ml of 2% acetic acid, was used to test the effect of some non-steroidal anti-inflammatory drugs. During the acute phlogistic reaction there is a variation of the cAMP/cGMP ratio, with a sharp increase of the cGMP concentration and a parallel decrease of the cAMP level in the exudate. Indometacin and 1-methyl-5-p-toluoylpyrrole-2-acetic acid (tolmetin, Tolectin), two non-steroidal anti-inflammatory drugs, injected intrapleurally 1 h before the acetic acid injection were able to reduce the inflammatory response, affecting the cAMP and cGMP ratio. That is there is an increase of the cAMP level and a decrease of the cGMP level.


Effect of 1,4-dimorpholino-7-(4-hydroxyphenyl)pyrido[3,4-d]) pyridazine [DS-511(4'-OH)] on the transepithelial transport of sodium and water and the permeability to urea in the toad urinary bladder. To elucidate the mechanism of the inhibitory action of 1,4-dimorpholino-7-phenylpyrido[3,4-d]pyridazine (DS-511) on water and sodium reabsorption at the renal tubules, the effect of DS-511 (4'-OH), which is similar in diuretic effect to but more water-soluble than DS-511, on the transepithelial transport of sodium and water and permeability to urea was studied in isolated toad urinary bladder. Application of DS-511(4'-OH) at concentrations above 2 x 10(-4) mol/l to the serosal side of the bladder depressed the transepithelial potential difference, short circuit current (SCC), and membrane conductance as well as the increased response of the SCC to arginine vasopressin (AVP) and cyclic AMP. The effect of DS-511 (4'-OH) applied to the mucosal side was delayed in onset and less pronounced. Neither serosal nor mucosal 10(-3) mol/l DS-511(4'-OH) depressed the increased response of the SCC to amphotericin B. 2 x 10(-4) mol/l DS-511 (4'-OH) applied to the serosal side did not affect osmotic water flow, but potentiated the increase in water flow caused by AVP. Basal urea permeability as well as the increase in urea permeability caused by AVP were depressed by serosal 10(-3) mol/l DS-511 (4'-OH). The results show that DS-511(4'-OH) has two actions, the depression of the transepithelial transport of sodium and urea, and the potentiation of the increased water permeability caused by AVP.


[The influence of pentifylline and theophylline on reaction kinetics on rat brain ATPase stimulatable by catecholamines (author's transl)]. The effects of 1-hexyl-3,7-dimethyl-xanthine (pentifylline, Cosaldon) and theophylline on rat brain Na+-, K+-, Mg2+ ATPase activities were investigated in in vitro experiments. It was established that pentifylline inhibits catecholamine sensitive ATPase in a dose dependent manner. Theophylline was without effect. Pentifylline also inhibited the Mg2+-dependent portion of Na+, K+-, Mg2+-ATPase. The effect of pentifylline on the kinetic parameters of Na+-, K+-, Mg2+-ATPase of synaptosomes was studied in detail. It was shown by a Lineweaver-Burk plot under the influence of pentifylline that the Michaelis constant (Km) increases from 1.0 x 10(-4) mol/l to 6.7 x 10(-4) mol/l. Km by norepinephrine stimulated ATPase decreases from 3.7 x 10(-4) mol/l to 2.9 x 10(-4) mol/l. In both experimental situations a decrease of maximal reaction velocity (Vmax) was observed. At high concentration of potassium in incubation medium the ATPase of synaptosomes was significantly more sensitive to pentifylline than at low concentration of potassium. The inhibition of ATPase by pentifylline was not influenced by the change in Na+/K+ ratio in the incubation medium. In all these experiments, theophylline used as a standard xanthine, was virtually without effect on the reaction kinetic of Na+-, K+-ATPase.


Studies on the mechanism of action of protacine (CR 604), a new non-steroidal anti-inflammatory agent. 3'-(4-[2-(1-P-Chlorobenzoyl-5-methoxy-2-methyl-indol-3-yl-acetoxy)-ethyl]-piperazin-1-yl)propyl-4-benzamido-N,N-dipropyl-glutaramate(+/-)dimaleate (protacine, CR 604), a new non-steroidal compound active on experimental inflammation, in vitro inhibits prostaglandin synthesis from arachidonic acid and platelet aggregation. When administered p.o., it prevents ex vivo platelet aggregation and in vivo arachidonate-induced thrombosis in rabbits. The cAMP levels of rat leucocytes are significantly reduced after treatment in vivo with protacine, even under maximum PGE-mediated stimulation. These effects, the inhibition of the proteolytic activity of trypsin and the fibrinolytic properties evidenced on recalcified plasma clots, contribute to explain the anti-inflammatory activity of protacine.


The effect of beta-pyridylcarbinol on lipoprotein lipids in primary type IIa hyperlipoproteinemia. The effect of long-term treatment over 6 months with beta-pyridylcarbinol on the lipoprotein lipids was investigated in 12 patients with primary type IIa hyperlipoproteinemia. VLDL, LDL and HDL were separated by preparative ultracentrifugation. There was a significant decrease of serum cholesterol and phospholipids. The normal serum triglycerides were unaffected. While VLDL and HDL lipids showed no signicant alterations, the LDL lipids decreased. The atypical lipid composition of the LDL was changed towards normal. Though there was a significant decrease of the "atherogenic" LDL/HDL-lipid ratios after 6 months treatment, beta-pyridylcarbinol did not result in a normalization of this ratio.


Comparison of high density lipoprotein and serum cholesterol levels in a European and an African community. Serum and high density lipoprotein cholesterol concentrations have been measured in 210 men and women in a London community and in a Nsukka community. Mean serum cholesterol concentrations were higher in London than in Nsukka. High density lipoprotein cholesterol was, however, higher in Nsukka than in London. Whereas the difference in serum cholesterol reached a statistical level when all the age groups were considered together, there was no statistical difference when the two populations were considered in terms of the young (20-39 year age group) and the middle aged (40-59 year age group). Difference in high density lipoprotein cholestrol, however, reached a statistical level in these two age groups.


Familial type I hyperlipoproteinemia caused by apolipoprotein C-II deficiency. A study was made on the clinical and biochemical features of siblings of patients with hyperchylomicronemia and its inherited relationship. It was not a case of the classical type of familial LPL deficiency, but of familial apolipoprotein C-II deficiency. The first patient with apolipoprotein C-II deficiency was reported by Breckenridge et al. and our patients provide the basis for the second report of this new disease. Our observations in this study strongly suggest that familial apolipoprotein C-II deficiency is transmitted by an autosomal recessive mode of inheritance and heterozygotes of this disorder have no abnormalities of plasma lipid and lipoproteins in spite of the reduced plasma apolipoprotein C-II.


Rate of exchange of esterified cholesterol between human plasma low and high density lipoproteins. In vitro incubations of human low density lipoproteins (LDL), high density lipoproteins (HDL) and lipoprotein-free plasma at 37 degrees for 6 h revealed no significant net mass transfers of esterified cholesterol from either lipoprotein fraction to the other. Observed transfers of esterified [3H]cholesterol from LDL to HDL must, therefore, have represented a process of molecular exchange between the two fractions. The time course of the esterified [3H]cholesterol transfers indicated that, from the point of view of the exchange process, the pools of esterified cholesterol in LDL and HDL were (mathematically) homogeneous, thus permitting a precise calculation of the rate of the exchange between the two fractions. In incubations of lipoproteins isolated from the plasma of 9 clinically normal human subjects, there was an hourly exchange of 8-12% and 7-31% of the pools of esterified cholesterol in LDL and HDL respectively, which, in molar terms, represented an exchange of 108-301 mumol/l plasma/h between the two fractions. The exchange of esterified cholesterol between human LDL and HDL is, therefore, rapid when considered in relation to other parameters of esterified cholesterol and lipoprotein metabolism.


A longitudinal study of the biological variability of plasma lipoproteins in healthy young adults. The fasting serum concentrations of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total triglyceride and apoprotein A1 were measured at intervals of 12-18 weeks for 60 weeks in 17 male and 11 female healthy young adults in order to assess the variability of these risk factors for coronary disease. No statistically significant seasonal changes were detected in any variable in either sex, although a progressive rise in apoprotein A1 concentration was observed. The coefficients of variation for random fluctuations with time were in the rank order: total cholesterol less than HDL-C less than apoprotein A1 less than LDL-C less than triglyceride. These differences were attributable to biological, rather than to methodological, factors. Within subjects, HDL cholesterol concentration varied inversely with triglyceride concentration and directly with apoprotein A1 concentration. The marked differences which exist in the biological variability of lipid risk factors for atherosclerosis need to be taken into account when making comparisons in epidemiological studies of the predictive powers of single on-entry measurements for future disease. Fluctuations of HDL-C with time appear to be related in part to variations in triglyceride-rich lipoprotein metabolism.


Why very low density lipoprotein levels are normal in familial hyperchylomicronaemia. The anomalous finding that very low density lipoprotein levels are relatively normal in patients with familial hyperchylomicronaemia has never been satisfactorily explained, particularly in view of the marked reduction or absence of peripheral lipoprotein lipase activity characteristic of this condition. I propose that the discrepancy between the plasma levels of the two triglyceride-rich lipoprotein fractions in these patients is due to the secretion by the liver of triglyceride in the form of chylomicron-like particles, rather than as very low density lipoprotein. The proposed "switch" in the spectrum of lipoproteins secreted by the liver is probably contingent upon the activity of the hepatic lipase present on the liver cell plasma membrane.


[A new method for determination of ATPase activity in tissue homogenates]. Authors describe a new method to determine Mg2+ and Na+K+ATPase activity on extremely small samples of tissue homogenates. They use specific inhibitors to discern between the two activities and a sensitive colorimetric method to dose Pi released from ATP, Lower limit of sensitivity is about 0,0100 micromoles of Pi/mg proteins/hour.


Effects of anaesthesia and surgery on plasma aldosterone concentration and renin activity in man. The effects of various anaesthetic agents and surgery on the plasma concentrations of aldosterone and on renin activity were investigated in 76 patients. The plasma concentration of aldosterone was increased markedly by 2.5 times the control value during 45 min of ether anaesthesia. It was increased also during halothane, methoxyflurane and enflurane anaesthesia. Spinal anaesthesia did not alter significantly the plasma concentration of aldosterone. Plasma renin activity and ACTH concentration were increased slightly during general anaesthesia.


Deoxyribonuclease activity found in Epstein--Barr virus producing lymphoblastoid cells. A deoxyribonuclease activity from Epstein--Barr (EB) virus producer lymphocyte cell lines which is correlated with viral production and which is not present in virus non-producer or negative lymphocyte cell lines has been purified 220-fold with 20% recovery and characterized. This nuclease copurifies through diethylaminoethylcellulose column chromatography with the EB virus induced deoxyribonucleic acid (DNA) polymerase in EB virus producer cells which was recently reported by this laboratory, but elutes as a separate peak of activity upon phosphocellulose chromatography. This nuclease activity has a sedimentation coefficient of 4.0 S, a strong divalent cation requirement, an alkaline pH optimum, and the ability to utilize both native and denatured lymphocyte DNA as substrate, reducing both to monophosphonucleosides.


Stereochemistry of the hydrolysis of the endo isomer of uridine 2',3'-cyclic phosphorothioate catalyzed by the nonspecific phosphohydrolase from Enterobacter aerogenes. The nonspecific phosphohydrolase from Enterobacter aerogenes (ATCC 13048) requires divalent metal ions for activity, since zinc present in the isolated enzyme can be removed by extensive dialysis against 8-hydroxyquinoline-5-sulfonate at pH 7.5 to yield an inactive enzyme which can be reactivated by addition of Zn2+, Cd2+, Co2+, Mn2+, or Ni2+; six ions of either zinc or cadmium can be incorporated into the inactive enzyme, and this incorporation of metal ion can be correlated with the regaining of activity (J. A. Gerlt, R. Dhesi, and H. C. Hemmings, unpublished experiments). The cadmium-reactivated phosphohydrolase catalyzes the hydrolysis of the endo isomer of uridine 2',3'-cyclic phosphorothioate (U greater than pS) to yield uridine 3'-monophosphorothioate as the major product. After enzymatic hydrolysis of the cyclic phosphorothioate in 19.8% H218O and chemical recyclization of the 18O-labeled acyclic phosphorothioates to yield a mixture of the endo and exo isomers of U greater than pS, 18O is found primarily in the exo isomer, as judged by examination of the 145.7-MHz phosphorus-31 nuclear magnetic resonance spectrum of the mixture. This observation indicates that the cadmium phosphosphohydrolase catalyzes hydrolysis of endo-U greater than pS with inversion of configuration, implying that the hydrolysis reaction proceeds by an in-line attack of water on the phosphorus.