[Morphometric analysis of ultrastructure of human synovioma cells].

Under examination were two gigantic-cell benign synoviomas and seven synovial sarcomas, including two metastatic tumors. The most specific ultrastructural criterion allowing a judgement of the degree of malignancy of the neoplasm under study is a parameter characterizing the predominance of ergastoplasmic protein synthesis over polyribosomal one.


[Helper viruses of adeno-associated virus type 4 replication].

In replication of adeno-associated virus type 4 (AAV-4) the helper function may be performed by a non-defective virus from the same group of parvoviruses (Kilham virus). The synthesis of AAV-4 antigen was observed in a pig embryo kidney cell line, SPEV, chronically infected with Kilham virus, strain RV-13, 45--52 passages. A one-day-old SPEV-Kilham culture was infected with AAV-4. The AAV-4 antigen was detected by immunofluorescence at 6, 8, 12, 18 hours, 2, 3, 4, and 5 days after inoculation. During the first 2--4 days after inoculation the AAV-4 antigen was found in the nucleus and perinuclear zone, later in the cytoplasm. A "new" helper virus for AAV-4 replication has been found: simiancytomegalovirus in human embryo fibroblast cell culture permissive for the helper virus. In the systems where AAV-4 replicates, its antigen can be detected in the nucleus and perinuclear zone by IF. AAV-4 did not replicate in a system insensitive to the helper virus or under non-permissive conditions: at the time, the AAV-4 antigen localized only in the cell cytoplasm was detected.


[Dacarbacinum (DTIC) and bronchogenic carcinoma (author's transl)].

A pilot study is reported on the results achieved with the cytostatic agent dacarbacinum (DTIC) on previously untreated, inoperable bronchogenic carcinomas which had been histologically and/or cytologically verified. Altogether there were 16 patients, i.e. 13 males and 3 females. DTIC monotherapy was administered to 14 patients; one patient received additionally vitamin A in high dosage and a further patient VP 16 (a podophyllotoxin preparation). The achieved results (a remission rate of 50%) justify the further application of this agent, whereby its relatively limited side effects as well as goood tolerability in elderly patients (over 70 years of age) are emphasized. The inclusion of the preparation in effective combinations is discussed.


Pneumocystis carinii, Toxoplasma gondii, Cytomegalovirus and the compromised host.

Pneumocystis carinii and Toxoplasma gondii are the two major parasitic protozoan pathogens in the immunocompromised host. Both organisms cause latent infection in humans and many animals. Cats are the definitive hosts for toxoplasmosis; the animal vector for pneumocystis (if any) has not been defined. Toxoplasma is an obligate intracellular parasite, whereas the available evidence suggests that Pneumocystis carinii exists primarily extracellularly. In compromised hosts, pneumocystis infection usually involves only lungs, whereas toxoplasma causes a generalized infection with encephalitis being the principal clinical manifestation. Both types of infection are treated with combinations of folate antagonists (trimethoprim or pyrimethamine with sulfonamide). Both parasites are associated with cytomegalovirus infection in immunosuppressed hosts, an association which may be due to symbiosis between parasites, or to an additive immunosuppressive effect of dual infection on the hosts.


[Characteristics of the response of mice with temporary immunodepression to the administration of live or inactivated vaccine against tick-borne encephalitis].

Mice with immunity defects caused by administration of cyclophosphane were inoculated with an inactivated vaccine or Langat virus (TP-21 strain) considered to be an approximate model of the vaccine against tick-borne encephalitis. In mice with depressed immune response, the asymptomatic infection caused by Langat virus turns into acute form and encephalitis with a fatal outcome develops in 86--100%. Pre-treatment with inactivated tick-borne encephalitis vaccine did not protect the animals with the immunity defect against neuroinfection induced by subsequent inoculation with Langat or tick-borne encephalitis virus. The experimental results suggest that the use of a live vaccine against tick-borne encephalitis in specimens with immunity defects is very dangerous because of the possibility of affection of the central nervous system.


Kinetic investigations of the autoxidation of adrenalin.

Generation rates of superoxide anions (O2-) by autoxidizing adrenalin at pH 9.5 were determined in solutions containing either superoxide dismutase (0.85 M-1 S-1) or hydroxylamine (0.0185 M-1 S-1) as competitive scavengers. The rate constants of O2- with adrenalin and hydroxylamine were calculated for neutral and alkaline solutions. The respective values were for adrenalin: 5.6 X 10(4) M-1 S-1, pH 7.8; 7.0 X 10(3) M-1 S-1, pH 9.5--and for hydroxylamine 5.9 X 10(4) M-1 S-1, pH 7.8; 3.4 X 10(4) M-1 S-1, pH 9.5. The effects of various competitors and O2- sources on the rate constants were compared.


[Prenatal diagnosis of malformations by ultrasound (author's transl)].

The significance of ultrasound diagnosis for the detection of fetal malformations will be demonstrated in 66 cases. The group includes hydrocephaly, anencephaly, microcephaly, synostosis praematura, cleft of the face, spina bifida, omphalocoele, Potter's syndrome, Down's syndrome, Klinefelter's syndrome, Vrolik's syndrome, triploidy, adenomatoide cystic malformation of the lungs, malformation of the heart and the limbs. Typical echographic findings are demonstrated. In regard to normal fetuses, head and trunk measurements will be analysed. Suspicious symptomes for the presence of malformations will be mentioned and also difficulties for the diagnosis of anencephaly and hydrocephaly. Furthermore typical findings in early pregnancy failure resulting from malformation are discussed.


Collagenase-induced experimental arthritis.

Rabbits were injected intraarticularly three times with 0.5 ml of 10, 50 or 100 mg% purified rheumatoid synovial collagenase or with identical amounts of trypsin. 18 hours after the last injection the collagenase-injected animals showed distinct cellular exudation into the synovial fluid and acute arthritis; one week later there was a decrease of the cell count in the synovial fluid and appearance of proliferative synovitis, while 3 weeks later there was no exudation and less chronic inflammation but distinct fibrosis of the synovium. A direct action of collagenase on the connective tissue components would seem to be responsible for these changes. No pathologic alteration of the cartilage was observed. Trypsin-injected control animals showed negligible cellular exudation and no pathologic alteration of the synovium.


[Comparative study of smallpox vaccines from strains B-51, EM-63 and L-IVP in a controlled epidemiologic trial. I. Reactogenic characteristics of the smallpox vaccines].

A comparative study of smallpox vaccines prepared from the l-ivp zm-63 and b-51 strains was carried out under conditions of controlled epidemiological trial. Children subject to planned vaccination were scarified with commercial batches of the preparations under study (with the same infectious activity). As a result of investigations it was found that in the group of those scarified with the vaccine from zm-63 strain strong local and catarrhal reactions and some disturbances of general condition were more frequent than in those scarified with preparations from the l-ivp and b-51 strain. Along with this, the temperature elevation of various intensity showed no statistically significant difference in the groups a vaccinated with the vaccines compared. Integral analysis of signs characterising different manifestations of reactigenic properties of the vaccines compared led to the conclusion that preparations made of zm-6 strain had greater reactogenic properties.


[Results of a study of acute paretic diseases in children according to material from a epidemiologic examination of the foci].

The authors carried out epidemiological examination of 160 foci of acute paretic diseases (APD) with serovirological examination of 181 patients and 413 contacts. Polioviruses were found to be etiologically responsible for the disease, as indicated by the results of serological examination in 37% of the patients, and by isolation of polioviruses (of the I and III types) in 4% of cases. As revealed, the vaccination scheme (periods and number of vaccinations) was not properly followed in 81.7% of those who contracted the disease. There were some differences in the characteristics of the foci in the child collective bodies and in families. Only individual cases of the disease were as a rule revealed in the familial foci, whereas in 9.6% of creches and kindergartens there were from 2 to 4 cases in each focus.


The role of cyclic nucleotides and prostaglandins in heart function.

A short review of the role of cyclic nucleotides and prostaglandins (PGs) in normal and pathological functions of the heart is given. Possible interrelationships of these two regulatory systems have been studied by using spontaneously beating rat atria preparations. Addition of noradrenaline (NA) to the incubate (1 . 10(-6) M) caused an increase in amplitude and frequency which was preceded and parallelled by an elevation of the tissue cAMP level. A transient increase in cGMP and PGE values was also seen. Propranolol (5 . 10(-6) M) abolished the increase in amplitude and frequency as well as in cAMP and PGE concentrations. Indomethacin (1 . 10(-5) M) inhibited the formation of PGE. The increase in cGMP was blocked by phenoxybenzamine. Interchange between beta- and alpha-receptors according as the temperature is lowered has been described earlier. Hypothermia (20 degrees C) had a positive inotropic effect on the atria and increased the tissue cAMP concentration. Loading of the atria caused an increase in cAMP without any effects on cGMP or PGs. Slight hypoxia did not change the cAMP or PG levels, but elevated the cGMP values. Arrhythmias induced by hypo- or hyperpotassemia did not modify the biochemical parameters measured. PGF2alpha (1. 10(-5) M) normalized the atrial rhythm and increased the amplitude without changing cyclic nucleotide or PG levels. PGE1 (1 . 10(-4) M) increased the amplitude of normorhythmic atria and the tissue concentration of cAMP. PGE2 was the only PG tested which stimulated the heart adenylate cyclase in vitro. There seems to be close but complicated relationships between cyclic nucleotides and PGs in the heart.


Effects of exogenous prostaglandin E1 on human plasma norepinephrine level and dopamine-beta-hydrolyxase activity.

Effects of exogenous PG E1 on the level of human plasma norepinephrine (NE), dopamine-beta-hydroxylase (DBH) activity, cAMP, cGMP, free fatty acids (FFA) as well as pulse rate and blood pressure were studied. Significant decreases of blood pressure and increases of pulse rate were observed after 20 min of the infusion of 0.05 microgram/kg/min of PG E1. The level of plasma NE increased by 174% (p less than 0.005) after the infusion. However, the increase of DBH activity was not significant. There was a tendency of increase of plasma level of FFA. These results suggested that the infusion of PG E1 caused an augmented sympathetic nervous activity due to systemic hypotension induced by PG E1.


On the histochemistry of prostaglandin-dehydrogenase.

Using PGF2alpha as substrate the PGDH-activity was localised in the male and in the female rat kidney by histochemical technique. The reaction is independent of the presence of NAD, but with NAD the reaction intensity is enhanced. The effectors phenazine methosulphate, Triton X 100 and Tween 80 showed no enhancing effect on the enzyme activity. In the rat kidney we have found the reaction product predominantly in the inner cortex (straight portion of the proximal tubules), however, sex dependent differences of PGDH-activity and localization are visible.


Effects of alpha- and beta-adrenergic blockers on binding of thyrotrophin to fat cell membrane.

Alpha- (phentolamine) and beta-adrenergic blocking agents (propranolol) and quinidine similarly enhance the specific binding of TSH to the guinea pig fat cell membranes over the concentration range of 2 X 10(-4) to 4 X 10(-3) M, increasing the binding affinity of TSH to the membranes. The percentage bound increased from 8% in the absence of agents to 32% (phentolamine), 29% (propranolol) and 24% (quinidine), respectively in the presence of these agents (1.5 X 10(-3) M). Each minimal detectable quantity of TSH was approximately 10 microU per tube in the presence of these agents (10(-3) M) as compared to 100 microU per tube in their absence. Both phentolamine and propranolol appeared to enhance the TSH binding to fat cell membranes through membrane-active, non-specific effects besides their alpha- and beta-adrenergic blocking activities.


Ovarian binding and intrinsic biological activity of desialylated human chorionic gonadothrophin (ASIALO-hCG).

The present study was undertaken to investigate the effect of desialylation on the intrinsic in vivo biological activity of hCG. To block the hepatic uptake of asialo-hCG we investigated asialo-orosomucoid. The biological activity of hCG and asialo-hCG was estimated with the ovarian ascorbic acid depletion assay (OAAD). The uptake studies were performed with [125I]hCG and [125I]asialo-hCG. Desialylation of hCG increased its hepatic uptake, almost completely abolished its ovarian uptake, and reduced its in vivo biological activity to less than 1%. Administration of asialo-orosomucoid caused a reduction of the hepatic uptake of asialo-hCG to the level observed for hCG. The ovarian uptake increased, but was lower than observed for hCG, due to a considerably higher renal accumulation of asialo-hCG. The in vivo biological activity of asialo-hCG was approximately 1/5 of the activity of hCG when both were tested in the presence of asialo-orosomucoid. To exclude the interference of renal accumulation, binding and biological activity were also investigated after bilateral nephrectomy. The uptake of radioactivity in the ovaries following injection of [512I]asialo-hCG and asialo-orosomucoid was then almost the same as observed after injection of [125I]hCG alone. When both preparations were now tested in the presence of asialo-orosomucoid, the in vivo biological activity of asialo-hCG appeared to be approximately 1/3 of the value obtained for hCG. It is concluded that the intrinsic in vivo biological activity of hCG is lowered by desialylation.


Myeloperoxidase activity and bactericidal function of PMN in iron deficiency.

In children with iron deficiency anemia, bactericidal capacity of polymorphonuclear leukocytes (PMN) and serum opsonic activity were studied. Nitroblue tetrazolium test (NBT), hexose monophosphate (HMP) shunt activation, and myeloperoxidase (MPO) activity of PMN of these cases were also examined. Bactericidal capacity and HMP shunt activation were found to be decreased in iron deficiency anemia ( p less than 0.001). MPO activity, NBT test, and serum opsonic activity were found to be within normal limits. After 1 1/2 months of iron therapy there was an improvement in bactericidal capacity and it returned to a normal level after 3 months of therapy.


Haemoglobin Barts in newborn Tanzanians.

Using both starch gel and cellulose-acetate electrophoresis is screening procedures, haemoglobin (Hb) Barts was detected in 11.08% of 325 cord blood samples from newborn Tanzanians. Red cell studies in these and in normals and a search for inclusion bodies of Hb H did not suggest alpha-thalassaemia. The mothers of these babies do not show any evidence of alpha-thalassaemia. It is suggested that the presence of Hb Barts in samples of cord blood is not due to the presence of alpha-thalassaemia in the Tanzanian population.


A controlled study of the effect of sulfinpyrazone on platelet survival and on platelet-bound 14C-serotonin release in patients with previous myocardial infarction.

Preliminary data obtained in the ambit of Anturan Reinfarction Italian Study (ARIS) show that, in postmyocardial infarction, reduced platelet survival time occurred in hyperbetalipoproteinemic patients treated with placebo, but not in the group of patients treated with sulfinpyrazone (interaction between treatment and lipemic level is at p approximately equal to 0.1). The sulfinpyrazone effect on platelet survival is probably related to the release reaction inhibition as suggested by our ex vivo results on platelet-bound 14C-serotonin release.


Cytochemical localization of acid phosphatase and adenosine triphosphatase in some avian mechanoreceptors.

Ultrastructural distribution of acid phosphatase and adenosine triphosphatase was studied in the receptor elements of HERBST and GRANDRY sensory corpuscles. Acid phosphatase activity was established in the elements of smooth and rough endoplasmic reticulum of perineural capsule cells, as well as in the secondary lysosomes of all cell types. Particular interest was paid on the activity of myelin-like dense bodies and some clear core vesicles belonging to the axoplasm of receptor nerve fibres. Adenosine triphosphatase activity was established on the membranes of receptor structures and pinocytotic vesicles. More deposits of electron dense material were localized on the axolemma of the non-myelinated portions of the receptor nerve fibres. The functional significance and importance of the both enzymes in the receptor structures was discussed.


Effect of norepinephrine on myocardial reactive hyperaemia in the isolated fibrillating dog heart.

The effect of intracoronary norepinephrine (NE) infusion in a dose of 2 microgram/min on postocclusion hyperaemic reactions following occlusions of 15 sec, 30 sec, 60 sec, and 120 sec was studied on isolated fibrillating, blood-perfused canine hearts. Basal perfusion pressure was kept at the level of 150 mm Hg; perfusion was done with constant pressure or constant volume. During administration of NE, basal flow and basal coronary conductance increased with both perfusion techniques. In the postocclusion period during administration of NE, peak conductance, maximum conductance, reactivity, mean transit time of reactive hyperaemia, postocclusive conductance area and repayment decreased. The differences found in postocclusion reactions in the control state originated from the different perfusion techniques disappeared when NE was infused. The phenomena may be explained by the stimulation of alpha and beta receptors as well as of myocardial metabolism by NE. The results support earlier views which ascribe a decisive role to vasoactive metabolites in the postocclusion reaction after prolonged occlusions. The decrease in reactive hyperaemia by NE may contribute to the development of angina pectoris.


Dissociation of the mitogenic and rosette-forming effects of PHA to stimulators of cyclic AMP levels.

The mitogenic and the spontaneous E-rosette stimulating effects of phytohaemagglutinin (PHA) were studied by examining their responses to an elevation of intracellular cAMP induced by pretreatment with aminophylline, adrenaline or PGE1 or by adding exogenous cAMP. Elevation of the intracellular cAMP levels of lymphocytes was found inhibitory to the mitogenic effect of PHA, while leaving the increased early and stable spontaneous rosette formation unaffected. It also failed to depress the PHA-induced slight increase in the proportion of total spontaneous E-rosette-forming lymphocytes. The responses of the two PHA effects under study to the elevation of the cAMP level are thus dissociated, suggesting that the two effects represent two different manifestations of lymphocyte activation, though these manifestations may not be independent of each other. It has been confirmed that PHA produces a shift primarily within the T-cell population toward the active and the stable E-rosette-forming subpopulation, while the increase in the total E-rosette-forming lymphocyte population is but slight. This may possibly involve the activation of the T-derived "null" cells unable to bind sheep red blood cells without PHA treatment.


Neurofibrillary degeneration in a case of quadriplegia and myoclonic movement.

A case of spastic quadriplegia with myoclonic movement was studied by both light and electron microscope. Argentophilic inclusions were found in a unique distribution involving the cerebral cortex, brain stem, cerebellum, and the spinal cord, including the motor neuron system. The fine structure of the inclusions consisted of accumulation of previously undescribed straight or wavy tubules approximately 120A wide devoid of periodic constriction.


Diagnostic evaluation of progesterone. Challenge test in amenorrheic patients.

Forty-one amenorrheic patients were grouped on the basis of presence or absence of withdrawal uterine bleeding following the intramuscular administration of progesterone. Ovarian volume, ovarian morphology with particular reference to presence or absence of follicles and state of follicular development, and steroidogenic function were investigated in each group. Most of amenorrheic patients with progesterone-induced uterine bleeding had relatively large ovaries with follicles of high developmental stage (tertiary-Graafian follicle) and responded to exogenously administered HMG and HCG with a rise in the 24-hour urinary excretion of total estrogens. In contrast, most of amenorrheic patients without progesterone-induced uterine bleeding had relatively small ovaries without follicles or with follicles of low developmental stage (primordial-secondary follicle) and did not respond to exogenous HMG and HCG. The results of the present study suggest that presence or absence of progesterone-induced uterine bleeding is closely correlated with the volume, morphology and steroidogenic function of the ovary in amenornorrheic patients. Thus, pathologic amenorrhea could be divided into two groups by utilizing the progesterone challenge test and this clinical categorization might be useful for the diagnosis and treatment of amenorrheic patients.


S-IgA cholera toxin and rotavirus antibody in human colostrum.

S-IgA antibodies against cholera toxin and rotavirus were assayed in 43 colostral samples by means of an enzyme-linked immunosorbent assay (ELISA). All specimens contained specific S-IgA antibodies against both antigens. Significant antibody titres to the antigens were demonstrated in almost all colostral samples.


Structural and functional abnormalities of the small intestine in infants and young children with rotavirus enteritis.

Structural and functional alterations in duodenal mucosa from 17 children with rotavirus enteritis were assessed. Structural changes were found in specimens from all patients. Patients with the most severe mucosal damage were more likely to require intravenous therapy to correct dehydration. Depression of one or more mucosal disaccharidases was found in 14 of 16 patients. Repeat duodenal biopsy three to eight weeks later in six patients showed marked improvement. The study clearly shows that rotavirus can cause a marked structural and functional lesion in the upper small intestine which is rapidly reversible.


17-Hydroxyprogesterone in normal children and congenital adrenal hyperplasia. Measurement in serum by radioimmunoassay after thin-layer chromatography.

Serum 17alpha-hydroxyprogesterone (17 OH-P) was measured by a specific radioimmunoassay technique combined with thin-layer chromatography. Normal values for children are less than 1.1 microgram/1 (less than 3.3 nmol/l)--corresponding to values found in the literature. In congenital adrenal hyperplasia (CAH) values up to several hundred microgram/l are found. The values rise after ACTH stimulation and are suppressed by decadrone or cortisone treatment. The rise in 17 ketosteroids and pregnanetriol in untreated CAH is relatively smaller (15--25 fold). This clinical sensitivity of 17 OH-P is thus valuable for the diagnosis of CAH (21 hydroxylase deficiency). Furthermore it is easier to take a blood sample than to collect urine for 24 hours. The usefulness in therapeutic monitoring is being studied.


Growth hormone response to prostaglandin E2.

C-AMP causes release of growth hormone from the pituitary. Prostaglandin increases pituitary adenylate cyclase and C-AMP and therefore acts like a growth hormone-releasing hormone. Using PGE2 (30 microgram per kilo body weight) given intravenously we have demonstrated that in no case was there a growth hormone response to insulin hypoglycaemia when PGE2 failed to evoke a response. However in 6 of 18 patients unresponsive to insulin hypoglycaemia a significant rise in growth hormone was obtained from PGE2. We argue that in these 6 patients hypothalamic unresponsiveness to hypoglycaemia must be operative whereas PGE2 acting directly at pituitary level is likely to have caused the release of preformed growth hormone from the pituitary. Administration of PGE2 does not cause hypoglycaemia but rather a slight rise in the plasma glucose level. Thus the risk of brain damage which is inherent in the insulin hypoglycaemia test is avoided.


Bilateral bone-marrow examinations in small-cell anaplastic carcinoma of the lung.

Bilateral bone-marrow examinations from the posterior iliac crests were routinely performed as a pretreatment staging procedure in 89 consecutive patients with small-cell anaplastic carcinoma of the lung. Bone-marrow involvement was found in 20 patients (22.5%), in 9 (10.1%) only on one side, and in 11 (12.4%) on both sides. Aspiration was found to be significantly superior to biopsy, being positive in 25 examinations as compared with 14 positive biopsies. Compared with unilateral bone-marrow examination the positive findings increased by approximately 30%. Bilateral bone-marrow examinations are recommended in patients in whom the detection of bone-marrow metastases will have therapeutic implications.


Influence of 4-aminopyrazolopyrimidine on morphology, synthesis of triglyceride and protein and their secretion in rat hepatocytes.

The influence of 4-aminopyrazolopyrimidine (4-APP) on morphology and on synthesis and secretion ability of isolated rat hepatocytes was investigated: 4-APP was found to inhibit both the synthesis and secretion of proteins. The synthesis of triglycerides was unaffected by 4-APP, while the secretion of triglycerides was markedly reduced. Transmission electron microscopy revealed that 4-APP induced morphological changes in the smooth membrane systems, the smooth endoplasmic reticulum and the Golgi apparatus. The possibility that 4-APP inhibits lipoprotein formation by inhibition of the apoprotein synthesis is discussed.


Possible role of thiamine in neuromuscular transmission.

Pyrithiamine (50 mg/kg), a thiamine antagonist, decreased the muscle twitches of the rat masseter muscle at stimulation frequencies above 1 Hz 40--80 min after an i.v. injection. The post-tetanic potentiation (PTP) induced by nerve stimulation of the masseter muscle was abolished by pyrithiamine. Administration of thiamine restored the muscle twitches at stimulation frequencies above 1 Hz and the PTP. The muscle twitches elicited by direct muscle stimulation were not affected by pyrithiamine treatment. The abolishment of the PTP was accompanied by a decrease in thiamine and thiamine-diphosphate. The pyruvate level in the blood was unchanged after pyrithiamine treatment. Oxythiamine, on the other hand, had no effect on the PTP but increased the pyruvate level in the blood. Fern extract which contains thiaminase I also abolished the PTP--an effect reversible by the addition of thiamine. The frequency-induced depression of the muscle twitches induced by pyrithiamine was similar to the effect of low doses of d-tubocurarine (8 microgram/kg). The results support the hypothesis that thiamine may play a role in neuromuscular transmission.


Effect of silicic acid on surface potential of erythrocytes.

A potential-generating effect of ortho-silicic acid on the surface potentials of erythrocyte membranes was demonstrated by a potentiometric method using platinum electrodes. With concentration rising from 10(-7) to 10(-4) the silicic acid reduced potential of washed erythrocytes which reached the final concentration a value about 36 mV. Higher concentrations of acid caused haemolysis.


Rapid increase of glycosaminoglycans in the aorta of hypercholesterolemic rats; a negative correlation with plasma HDL concentration.

Rats were kept either on a standard laboratory diet or a high cholesterol, olive oil diet for periods ranging from 1 day to 22 weeks. The effect of the high cholesterol, olive oil diet on the concentrations of cholesterol, glycosaminoglycans (GAGs) and collagen in aortic intima-media, were studied and the developing hyperlipidemia was characterized. The concentration of cholesterol in rat aorta was increased after 22 weeks' high cholesterol, olive oil diet, while collagen concentration was not affected. On the contrary, the concentration of aortic sulphated GAGs was significantly increased already after one week's high cholesterol, olive oil diet. The diet increased the formation of a cholesterol-rich very low density lipoprotein (VLDL), decreased high density lipoprotein (HDL)-associated cholesterol and phospholipids, but had virtually no effect on low density lipoprotein (LDL)-lipids. The concentrations of VLDL-cholesterol and -phospholipids showed positive correlations with the concentration of aortic GAGs (r = 0.89 and 0.83, respectively, P less than 0.05 for both). Stronger (negative) correlations were found between aortic GAGs and HDL-cholesterol and -phospholipids (r = 0.94 for both, P less than 0.01) suggesting that HDL may have a role in the control of arterial sulphated GAG concentration.


The effect of ACTH on the arterial blood pressure, heart rate and blood histamine level in rabbits.

A rapid blood pressure fall, persisting for over one hour, was observed after ACTH injection. The heart rate was initially increased and returned to its basal level within one hour. On the other hand, the level of histamine was significantly increased only in the 60th minute of the experiment. The results obtained showed that a possibility of an indirect ACTH action through an effect of histamine metabolism cannot be excluded.


Lack of specific prostaglandin antagonistic effects of 7-oxa-13-prostynoic acid on ovarian metabolism in vitro.

Prostaglandins (PGs) have been proposed to function as obligatory intermediates in the action of luteinizing hormone (LH) (Kuehl et al. 1970). This based on experiments on isolated mouse ovaries where the substance 7-oxa-13-prostynoic acid (7-oxa-13-PA) was found to block the effect of both PGE1 and LH on ovarian cyclic AMP formation and progesterone synthesis. We have studied whether 7-oxa-13-PA acts as a PG antagonist also on the isolated rat ovary, where PGs of the E type have many effects. Prepubertal rat ovaries were incubated up to 4 h in modified Krebs bicarbonate buffer containing glucose with PGE1 and 7-oxa-13-PA, alone and in combination. PGE1 stimulated the uptake of the non-utilizable amino acids alpha-aminoisobutyric acid (AIB) and cycloleucine in an equally wide dose-range as has earlier been found for the stimulation of lactic acid and cyclic AMP production as well as protein synthesis. 7-oxa-13-PA alone, in concentrations exceeding 25 microgram/ml, stimulated lactic acid production, but inhibited the incorporation of labelled leucine and the uptake of AIB. The uptake of cycloleucine was slightly stimulated. 7-oxa-13-PA did not antagonize the PGE1 effect on lactic acid production. 7-oxa-13-PA diminished the PGE1 effect on the uptake of amino acids and the incorporation into protein, but only in concentrations where it in itself inhibited the protein synthesis. These results show that 7-oxa-13-PA does not act as a PG-antagonist in the prepubertal rat ovary, since it cannot block the effects of PGs without having marked inherent effects.


Effect of vasopressin and oxytocin perfusion of the cerebral ventricles on evoked tongue jerks.

The effect of increased concentration of vasopressin and oxytocin in the cerebrospinal fluid on the excitability of the hypoglossal nerve nucleus was investigated. The experiments were carried out on rats under chloralose anaesthesia. Retractory jerks of the outstretched tongue were evoked by supra- or infraorbital nerve stimulation during perfusion of the cerebral ventricles with McIlwain-Rodnight solution. The solution contained synthetic arginine vasopressin 0.05 U/ml or synthetic oxytocin 0.05 U/ml. Perfusion of the ventricles with vasopressin increased and perfusion with oxytocin decreased the evoked tongue jerks.


On the distributive patterns of phosphatases and non-specific esterase and their functional aspects amongst the constituents of the photoreceptors of the Indian Medicinal leech, Poecilobdella granulosa (Savigny, 1822).

In the present contribution the distributive pattern of phosphatases and non-specific esterase amongst the constituents of the leech eye has been revealed and the significance of the enzymatic locales in relation to the visual processes has been discussed. All the phosphatases studied demonstrate absence of the activity in the lens. Activity of the phosphatases, however, is restricted to the plasma membrane and associated cytoplasm to varying degrees. The significance of the phosphatases in the nerve impulses and transmission has been highlightened. The observation of considerable significance relates the demonstration of a high degree of non-specific esterase activity in the lens, which has defied all the phosphatases. Another observation relates to the positive activity of 5-nucleotidase in the group of axons constituting optic nerve. The significance of this observation has also been discussed.


Electroencephalographic investigations in sellar tumours, with special regard to different methods of operative treatment.

Sixty-six patients with tumours in the sellar region were examined. All were operated on either by the transfrontal or the transsphenoidal route. Pre- and postoperative longitudinal electroencephalographic investigations were performed. Preoperative electroencephalograms showed a normal frequency content in cases of intrasellar tumours or those reaching the chiasma. Nearly all cases had irregularities in the temporal regions. Tumours compressing the third ventricle had slower average frequencies and a general slowing in all cases. Besides these alterations unilateral delta waves or bitemporal dysrhythmic groups were sometimes found. A connection between extension of the tumour and its histological nature could not be found, but the operative approach influenced the electroencephalographic disturbances enormously. After a transfrontal approach and removal of the tumour the electroencephalogram was unchanged. Sometimes a mild transient bitemporal slowing was present. But after a transfrontal operative approach a general slowing was common, usually with focal marked slow activity in the right fronto-temporal area.


Medical registers.

Registers are records in which information is precisely noted. Their form owes much to William Farr, who headed the General Register Office in Britain beginning in 1839. Medical registers may be designed for various purposes: preventive medical purposes; specific diseases; treatment, aftercare, and at-risk listings; skills and resources indices; prospective studies; and specific information purposes. Entries are correctly diagnosed consistently updated, and derived from multiple overlapping sources in well-defined populations. In Israel, a national neurologic disease registry was established in 1969 based on the diagnoses of discharge of all hospitalized patients. This registry has yielded valuable epidemiologic information on a large number of neurologic disorders.


Neurologic complications of diabetes mellitus: transient ischemic attack, stroke, and peripheral neuropathy.

The incidence of TIA, stroke, and neuropathy was studied in a community-based maturity-onset diabetic population. The frequencies of TIA and stroke were increased in maturity-onset diabetic patients as compared to the population of Rochester, Minnesota. The median age of occurrence of TIA and stroke in diabetics was 74 years, not significantly different from that in non-diabetics. Diabetic patients with hypertension at the time of diagnosis of diabetes mellitus had an increased frequency of TIA and stroke. Control of hypertension and/or diabetes mellitus was associated with a decreased frequency of TIA or stroke. Obesity, clinical coronary heart disease, and an abnormal electrocardiogram at the time of diagnosis of diabetes mellitus were not associated with a significantly increased frequency of TIA or stroke. The most common type of peripheral neuropathy in diabetes mellitus was distal polyneuropathy. Mononeuropathy and autonomic neuropathy were much less frequent. The frequency of distal polyneuropathy increased with the duration of diabetes mellitus. The frequency of neuropathy was increased in patients with poor control, reemphasizing the importance of diabetic control in the prevention of diabetic complications.


Neurologic complications of acute and chronic renal failure.

It is obvious from the preceding discussion that neurologic complications contribute significantly to morbidity and mortality in patients with chronic and acute renal failure and that dysfunction of the nervous system continues to occur even in those patients receiving successful renal transplants. In an effort to find the causes of these disorders, a large cooperative study of patients with renal failure will be needed to supply valid and statistically significant data that can be used to test the hypotheses suggested by this and other review papers. At the present time, the data are limited, because in almost all instances small and biased series of patients have been used, and basic criteria of disease have not been standardized. It will be a large task to correct these deficiencies, but it is definitely one that should be undertaken in the near future.


Unspecific and specific stimulating effects of 3',5'-cyclic nucleotides (cGMP, cAMP, cIMP, cCMP) on the in vitro biosynthesis of proteochondroitin-4,-6-sulfate and on other anabolic processes from calf rib cartilage.

With cartilage slices from calf ribs, cGMP as well as cAMP accelerate dose-dependently and specifically label rates of Ch-4-,-6-S protein; they slightly elevate rates of anaerobic glycolysis dose-independently and unspecifically, similar to their 5-monophosphate compounds. cAMP, but not cGMP, slightly stimulates labeling of total protein dose-dependently. Guanosine and adenosine (as well as adenine) accelerate more significantly all three anabolic processes in the order Ch-4-,-6-S protein formation greater than or equal to total protein labeling greater than anaerobic glycolysis. Acceleration of some of the processes rises further after adding theophylline or SQ 20.009, depending on the nucleoside used. diBu-cAMP (but not 8-Br-cAMP) stimulates the three processes more than cAMP; diBu-cGMP and 8-Br-cGMP alone increase the labeling rates of protein more than cGMP, cCMP and cIMP slightly accelerate at least one of the three processes dose-independently and unspecifically, similar to their 5-monophosphate compounds. cUMP was almost inactive. The results point to specific and unspecific effects of cGMP similar or different to those of cAMP.


Protective action of amiodarone against ventricular fibrillation in the isolated perfused rat heart.

The pretreatment of rats with amiodarone for 2 minutes to 3 weeks before the excision of their hearts caused a dose-related decrease in heart rate and an increase in the ventricular fibrillation threshold both before and after coronary arterial ligation. Similarly, amiodarone decreased the incidence of ventricular premature extrasystoles, ventricular tachycardia and fibrillation during the period of regional ischemia after coronary arterial ligation and also after reperfusion of the ischemic myocardium. There was no evidence of a metabolic protective effect on ischemic myocardium because tissue high energy phosphate content decreased to a similar extent in ischemic myocardium from control and amiodarone-treated rats. Instead, the protective effect of amiodarone against fibrillation was accompanied by attenuation of the increase in tissue cyclic adenosine monophosphate in ischemic myocardium after coronary arterial ligation. It is proposed that amiodarone exerts a potent antifibrillatory effect by decreasing tissue cyclic adenosine monophosphate in ischemic myocardium.


Leucocyte function in iron-deficiency anemia.

Production of hydrogenperoxide and myeloperoxidase-mediated iodination, by leucocytes was studied in normal subjects and subjects suffering from iron-deficiency anemia. The leucocytes obtained from severe cases of iron-deficiency anemia showed decreased myeloperoxidase-mediated iodination. Defective myeloperoxidase-mediated iodination can explain the basis for the leucocyte microbicidal defect in iron deficiency anemia.


Scientific advance in acupuncture.

In this paper we present scientific advance in acupuncture based on electrical, neurophysiological, biochemical and therapeutic studies made inside and outside China since the early fifties. New modalities other than traditional needling techniques notably developed in Japan, France and Germany are described. For future prospects, efforts by making use of systems approach, field-body interaction, self-defense self-strategy and circadian rhythm are likely to produce great fruition in medicine. Possibilities of making advances in "three P's" medicine (preventive medicine, precision medicine and people medicine) by acupuncture are discussed.


Thymectomy does not affect tumour rejection and cytophilic activity in a murine allogeneic lymphoma model.

An experimental model was devised in which an AKR lymphoma is conditioned to grow in BALB mice leading to reproducible tumor incidence which makes tumor-bearing (progressor) and tumor-rejecting (regressor) animals simultaneously available. The object of this paper was to determine the effect of neonatal thymectomy (xT) on allogeneic tumor incidence and on anti-tumor cytophilic activity. The latter was determined by the adherence of lymphoma cells to guinea pig peritoneal macrophages previously incubated with preheated mouse serum. The results obtained, in 2-3 month old animals, show no difference in lethal tumor incidence between xT and intact mice, 36% (14/39) vs. 39% (14/36). Neither did xT alter the significant increase in cytophilic antibodies detected in regressor serum, 115 +/- 15 (S.E.) vs. 106+/- 22 0/00 macrophages bearing tumor cells as compared to control values in either xT or normal serum, 53 +/- 3 vs 52 +/- 3. This background cytophilic activity was not significantly altered in progressor serum of either xT or intact mice, 36 +/- 5 vs 65 +/- 6. The specificity of the antitumor cytophilic antibodies was determined by the negative results obtained when a different tumor was used as target cell. It can be concluded that ant-tumor cytophilic antibodies are detectable in regressor but not in progressor serum. Thymectomy in this model does not alter either in vivo tumor incidence or humoral cytophilic activity. Since no thymic remnants were encountered at autopsy, it is postulated that AKR lymphoma cells, which proved to be neoplastic T cells, are capable of rendering T-immunocompetent a thymectomized allogeneic BALB host.


Adenosarcoma of the uterus: ultrastructural observations.

Müllerian adenosarcoma has been recently described as a distinctive biphasic uterine tumor with benign epithelial and malignant stromal components. The lesion belongs generally to the group of mixed müllerian tumors, but the benignity of the epithelium separates it from the other neoplasms of this type. This entity usually affects postmenopausal patients who follow a protracted course. This paper reports a case fulfilling the original clinical and pathologic description of Clement and Scully and adds ultrastructural observations. Electron microscopic examination confirmed the optical impression of a biphasic neoplasm with benign epithelial and malignant stromal components. The epithelium resembled proliferative endometrium, and the stroma had features of endometrial stromal sarcoma or mixed müllerian tumor. The findings of stromal resemblance to endometrial stromal sarcoma or mixed müllerian tumor and the presence of a variety of müllerian epithelia support the proposed müllerian derivation of this lesion.


Neuropsychiatric problems in mixed connective tissue disease.

A group of 20 patients with mixed connective tissue disease, followed for up to five years, was found to have a 55 per cent incidence of neuropsychiatric problems. An aseptic meningitis-like syndrome was the most common presentation and was rapidly responsive to corticosteroid therapy. Other findings were psychosis, convulsions, peripheral neuropathy, trigeminal neuropathy and cerebellar ataxia. An abnormal cerebrospinal fluid was found in five patients; mild pleocytosis, an increased protein content and a first phase colloidal gold curve were the main abnormalities. These neuropsychiatric problems have not been a cause of mortality in this group of patients with mixed connective tissue disease.


Leukocyte myeloperoxidase deficiency and diabetes mellitus associated with Candida albicans liver abscess.

Reported here is a case of hereditary myeloperoxidase deficinecy in a diabetic patient suffering from a Candida albicans liver abscess. Peroxidase activity is completely absent from the neutrophils and monocytes although it is present in the eosinophils. The different forms of myeloperoxidase deficiency are discussed.


Influences of an analog of the neuropeptide ACTH 4--9 on mentally retarded adults.

In a double-blind procedure, 24 mentally retarded adults received 0 mg, 5mg, or 20 mg of an analog of the neuropeptide ACTH 4--9. Following treatment with peptide, the subjects were given the Trails B Test (from the Halstead-Reitan Neuropsychological Battery), the Peabody Picture Vocabulary test, the Benton Visual Retention Test, a concept-formation task, and a standard orienting sequence. The results of the behavioral tests suggested that attentional processes were enhanced in subjects treated with the peptide. The present study, in conjunction with another investigation using the neuropeptide ACTH/MSH 4--10 (Sandman, George, Walker, Nolan & Kastin, 1976), indicates that attentional deficits in mentally retarded adults, traditionally assumed to be irreversible, may be influenced by treatment with fragments of the neuropeptides ACTH and MSH.


Recurrent herpes simplex labialis: viral replication and clinical course.

The purpose of this study was to provide additional information on the clinical course and viral replication kinetics of herpes simplex virus (HSV) in recurrent herpes labialis. Data were obtained on 20 subjects who were followed for five days from the first day of the lesion. HSV was isolated in 17 subjects (85%); 75% were positive on day 1 of the lesion. Median HSV titer on day 1 was 1.7 x 10(3)/0.2ml. Isolation rates and titers dropped sharply and virus could no longer be detected by day 4. The clinical course both in terms of the frequency and the severity of the symptoms paralleled the kinetics of viral replication. Thermography delineated location of subsequent lesions in three early symptomatic patients who were studied at the time when no visible lesions were observed. These data are felt to provide useful background information for future studies on the efficacy of topical antiviral agents.


Infectious mononucleosis and related syndromes.

The "mono syndrome" is an acute febrile disease of older children and young adults which involves the lymphatic system and is characterized hematologically by the presence of 50% or more lymphocytes and monocytes and 10% or more atypical lymphocytes. Epstein-Barr virus (EBV) causes over 90% of the syndrome, cytomegalovirus (CMV) about 5% to 7%, and Toxoplasma gondii less than 1%. Viral hepatitis, adenovirus, rubella, and herpes simplex are rare causes. Only EBV produces classical heterophile-positive infectious mononucleosis. This article reviews the epidemiological and clinical features of this syndrome.


The hyperlipidemia of pregnancy in normal and complicated pregnancies.

Alterations in the concentrations of the cholesterol and triglyceride moieties of lipoproteins separated by ultracentrifugation and precipitation methods were studied at frequent intervals throughout pregnancy and the puerperium in a group of 43 women. The plasma cholesterol concentration rose on the average by about 50 per cent, the major increase occurring in the second trimester. The plasma triglyceride concentration rose threefold, reaching its peak during the third trimester. All major lipoproteins participated in these changes: in very-low-density lipoproteins, both lipids rose in proportion to the ratio in nonpregnant women, but in low-density and high-density lipoproteins, the ratio of triglyceride to cholesterol rose. The triglyceride enrighment in low-density lipoproteins reflected the inclusion of intermediate-density lipoproteins (d 1.006 to 1.019). The occurrence of hypertension or pre-eclampsia led to a further increase in lipids in very-low-density lipoproteins. Hypercholesterolemia was greatest in women with pre-existing hypercholesterolemia, and women in the third pregnancy showed higher plasma cholesterol concentrations than women in the first pregnancy. Both cholesterol and triglyceride concentrations decreased significantly within 24 hours of delivery and this was reflected in all lipoproteins. However, while triglyceride levels continued to decrease rapidly returning to nonpregnant levels during the puerperium, cholesterol in low-density lipoprotein remained elevated for at least six to seven weeks post partum.


Carpal radiographs in orthodontic treatment.

The pubescent period offers the best opportunity to accomplish the objectives of orthodontic treatment in the shortest time. Treatment during the period of maximum growth activity reduces some of the uncertainties associated with early treatment. Carpal radiographs can be used as a guide to determine the onset of the adolescent growth spurt and hence provide the clinician with a diagnostic and planning aid to optimize treatment efficiency.


Antiphosphaturic action of 25 (OH) vitamin D3 in experimental Fanconi syndrome.

Renal handling of phosphorus was studied in the following groups of parathyroidectomized rats with maleate-induced Fanconi syndrome: 1) 6 rats receiving intravenous parathyroid hormone, 2) 6 rats receiving intravenous dibutyryl cyclic AMP (DBcAMP), 3) 6 rats undergoing volume expansion with saline, 4) 12 rats receiving intravenous 25 (OH)vitamin D3, 5) 12 rats with acute hypercalcemia induced by intravenous CaCl2, 6) 6 rats with phosphate deprivation, and 7) 6 rats receiving intravenous calcitonin. Parathyroid hormone and calcitonin failed to increase the urinary excretion of both cAMP and phosphorus. Likewise, DBcAMP failed to increase the urinary excretion of phosphorus. Extracellular volume expansion and hypercalcemia (serum calcium 12.9 +/- 0.7 mg/100 ml) did not alter the tubular reabsorption of phosphorus. In phosphate-deprived animals, the fractional excretion 0.16 +/- 0.05 (mean +/- SE) was lower than that in the control animals (maleate-treated without phosphate depletion), 0.46 +/- 0.04 (P less than 0.001). 25 (OH)vitamin D3 decreased the fractional excretion of phosphorus from 0.39 +/- 0.03 in the control (maleate-treated not receiving 25 (OH)vitamin D3) to 0.23 +/- 0.02 (P less than 0.001) in the experimental animals. The present study demonstrated an antiphosphaturic effect of 25(OH)vitamin D3 in experimental Fanconi syndrome; the mechanism of this action is not well understood.


Relationship between PAH transport and Na-K-ATPase activity in the rabbit kidney.

The relationship between Na-K-ATPase activity and p-aminohippurate (PAH) transport was examined in renal cortical slices of mixed-breed rabbits by using ouabain to vary the level of Na-K-ATPase activity. Ouabain increased passive PAH uptake, measured in the presence of probenecid. Slice homogenate Na-K-ATPase activity was inhibited by 50% with about 2 X 10(-6) M ouabain. When the concentration was 10(-5) M or above, active PAH uptake was inhibited. Lower concentrations (10(-7) and 10(-6) M) of ouabain stimulated active PAH uptake. Acetate stimulated PAH uptake when medium ouabain was less than 10(-4) M. Ouabain, 10(-5) M and higher, caused intracellular K+ to fall and Na+ to rise. The changes in active PAH uptake observed with ouabain correlated much better with changes in intracellular K+ or Na+ levels (P less than 0.01) than with changes in slice homogenate Na-K-ATPase activity. Vmax for active PAH uptake decreased with 5 X 10(-5) M ouabain, and tended to increase with 10(-7) M ouabain. Ouabain did not alter Km for active PAH uptake. The data support the hypothesis that there is a functional link between active PAH transport and Na-K-ATPase activity. This linkage may be an indirect one mediated by changes in intracellular cation concentrations.


Modulation of adrenergic transmission by angiotensins in the perfused rat mesentery.

Isolated rat mesenteric arteries perfused with a modified Krebs solution were utilized to study the effects of angiotensin II (AII), angiotensin III (AIII), and [des-Asp1-Arg2]AII on adrenergic transmission. Angiotensin II potentiated vasoconstrictor responses to both sympathetic nerve stimulation and to exogenous norepinephrine, whereas AIII and [des-Asp1-Arg2]AII potentiated vasoconstrictor responses to exogenous norepinephrine only. When the responses to exogenous norepinephrine were compared, the order of agonist potency was AIII greater than AII greater than [des-Asp1-Arg2]AII. The potentiation of sympathetic nerve stimulation by AII was inhibited by simultaneous administration of AIII (25%), [des-Asp1-Arg2]AII (51%), [Sar1-Ile8]AII (83%), and (Ile7)AIII (80%). The potentiation of exogenous norepinephrine by AII, AIII, and [des-Asp1-Arg2]AII was inhibited by [Sar1-Ile8]AII (110%, 113%, and 108%, respectively) and by [Ile7]AIII (50%, 64%, 91%, respectively). We conclude that AII possesses the capacity both to increase norpinephrine release during sympathetic nerve stimulation and to decrease norepinephrine reuptake, whereas AIII and [des-Asp1-Arg2]AII decrease norepinephrine release and reuptake. Also, under conditions of increased N-terminal degradation of AII, blockade of norepinephrine reuptake would be increased while the release of norepinephrine by nerve stimulation would be reduced.


Development and regression of exercise-induced cardiac hypertrophy in rats.

Adult female rats were exercised by daily swimming. All the increase in heart weight induced by the exercise occurred within 14 days and averaged 30%. The half times of the increases in heart weight and total protein content were about 4.5 days, whereas that of cytochrome c, which was used as a mitochondrial marker, was 6.5 days. The total amounts of DNA and of hydroxyproline in the heart, which were used to evaluate the degree of connective tissue hyperplasia, increased only slightly (8% and 10%, respectively). Other animals were subjected to the same swimming program for 21 days. Groups of rats were killed at various time intervals after stopping exercise. Heart weight, total protein content, and total cytochrome c content decreased rapidly initially, with 60% of the total regression of hypertrophy occurring during the first week. Thereafter, heart weight fell more gradually toward the sedentary control value. The hydroxyproline content of the heart, which was increased 10%, did not decrease during the regression of the hypertrophy.


Swine influenza vaccine program in the community: acceptability, reactions and responses.

The operation of the national swine influenza immunization program was observed in the community of Tecumseh, Michigan. The purpose was to determine acceptability of a parenteral vaccine intended for the general population. Participation of the residents was excellent. More than 64 per cent of eligibles were vaccinated; this figure resembled that of the rest of the local area, but not the national figures nor a suburban area of Michigan where a similar study was carried out. Sore arm after vaccination was reported most frequently in younger female participants; however, sore arm was accepted as part of the process of vaccination and not considered a reaction by most. Such perceived reactions were not as commonly reported as in the large suburban area. Antibody response to the vaccine was excellent, not only in terms of antibodies to the swine virus itself but also in terms of cross reactive antibodies to the emergent H1N1 (Russian) influenza strain. As a result of the program, many vaccine recipients in the 25 to 49 year age groups are protected against this new epidemic virus.


Ultrastructural changes in the rat metrial gland in the latter half of pregnancy.

The ultrastructural changes occurring in the metrial gland in the latter half of pregnancy in the rat have been studied. Typical palely stained granulated cells are present in the metrial gland up to day 20 but many granulated cells show variations in appearance which may be associated with degeneration. In some the cytoplasm is more darkly stained and such cells often have apparently empty areas of cytoplasm adjacent to the granules. From day 14 onwards many areas of the gland show cellular debris, apparently resulting from lysis of the granulated cells. However, occasionally normal granulated cells are present in blood vessels and have been observed apparently penetrating the vascular endothelium. A variety of changes was also noted in the stromal cell population. Inclusions became apparent in many of these cells; some of these consisted of lipid while others resembled granules from the typical granulated cells. Other cells with numerous inclusions appeared to be macrophages. Lysis of granulated cells in situ is compatible with suggestions that the metrial gland produces a holocrine secretion, though some normal granulated cells enter blood vessels. The stromal cells may have a phagocytic role in late pregnancy but evidence for this was inconclusive.


Susceptibility to rhinovirus infection in chronic bronchitis.

The role of rhinoviruses in chronic bronchitis was examined serologically in the community of Tecumseh, Michigan. Five different rhinovirus types had been identified by isolation to be present in the community during the time of the study. They were tested against serums obtained from persons with chronic bronchitis and matched control subjects. Both males and females with chronic bronchitis had more frequent infections than did the control subjects; for males, the differences were significant. The higher infection rate among the bronchitic subjects was not related to their being deficient in antibody at the start of the study. It was concluded that persons with chronic bronchitis appear to be more susceptible to rhinovirus infection, as they are to other viral pathogens.


Phosphatidate phosphohydrolase activity in porcine pulmonary surfactant.

The composition and enzymatic activity of porcine surfactant obtained by lung lavage was examined. The ratio of phospholipid to protein was found to be 13 mg per mg. Phosphatidate phosphohydrolase (PAPase) activity was present in lavage surfactant, lamellar bodies isolated from porcine lung tissue, and lamellar bodies isolated from human amniotic fluid. The optimal pH of PAPase in surfactant in lamellar bodies was 6.0. Cholinephosphotransferase activity was also present gradient centrifugation of amniotic fluid, the highest PAPase specific activity was found in the fraction that banded at the 0.65 M sucrose interface, similar to lamellar bodies obtained from porcine lung. From these results, we conclude that the enzyme PAPase remains closely associated with the surface active lipids during the process of storage and secretion of surfactant.


Regulation of airway smooth muscle tone in sleeping dogs.

We examined the influence of sleep state on airway smooth muscle tone in 4 unanesthetized dogs that were trained to sleep in the laboratory. The dogs had been prepared with a permanent side-hole tracheostomy and bilateral cervical vagal loops. During the studies, the dogs breathed through a cuffed endotracheal tube inserted through the tracheostomy. To monitor changes in tracheal smooth muscle tone, we measured the pressure in the water-filled cuff of the endotracheal tube. The technique was validated by examining changes in cuff pressure after administration to the dogs of a series of chemical agents and physiologic stimuli known to constrict or relax tracheobronchial smooth muscle. Sleep state of the dogs was determined by behavioral, electroencephalographic, and electromyographic criteria. During quiet wakefulness, tracheal smooth muscle tone was stable. With the onset and progression of sleep through the nonrapid-eye movement stages, airway smooth muscle tone relaxed (decrease in cuff pressure of 20 to 40 cm H2O), reaching a new steady level during slow-wave sleep. In contrast, during rapid-eye-movement sleep, tracheal smooth muscle tone fluctuated markedly and erratically, as reflected by changes in cuff pressure as large as 90 cm H2O. Partial blockade of the vagus nerves, by cooling the exteriorized cervical vagal loops, decreased or abolished the fluctuations in tracheal smooth muscle tone during rapid-eye-movement sleep at temperatures that did not abolish resting tone, demonstrating that the changes in tone during rapid-eye-movement sleep were related to variability in neural control of airway smooth muscle.


Oral angiotensin-converting enzyme inhibitor in long-term treatment of hypertensive patients.

The antihypertensive effect of the orally active angiotensin-converting enzyme inhibitor captopril (SQ 14225) was assessed in 22 hypertensive patients of whom 17 were followed for periods ranging from 1 to 7 months. Of these, eight had essential hypertension, eight had renovascular hypertension, and six had hypertension associated with chronic renal failure. Blood pressure decreased markedly in all patients, including those with low renin levels. Nevertheless, the magnitude of blood pressure reduction correlated with the base-line plasma renin activity (r = 0.58, P less than 0.01). Increasing the dose of captopril from 25 to 200 mg did not enhance the amplitude of the antihypertensive effect but did increase its duration. Patients' blood pressure remained well controlled and free of side-effects with a maximal daily dose of up to 200 mg by mouth twice daily. Despite the blood pressure reduction, sodium excretion tended to increase, probably because of reduced aldosterone secretion. There was no evidence of orthostatic hypotension, and no escape from the antihypertensive effect was observed. These results indicate that chronic inhibition of the angiotensin-converting enzyme with an orally active compound offers a new, efficient, and well-tolerated approach to the treatment of hypertension.


Cholesterol in the prediction of atherosclerotic disease. New perspectives based on the Framingham study.

Prospective data at Framingham and elsewhere have shown conclusively that risk of coronary heart disease in persons younger than age 50 is strikingly related to the serum total cholesterol level. Within so-called normal limits risk has been found to mount over a five-fold range. The impact has been found to be augmented by other risk factors. The contribution of the serum total cholesterol to risk has also been found to be determined by its partition in the various lipoprotein fractions. A relatively large amount of cholesterol in the low-density lipoprotein fraction is atherogenic, whereas that in the high-density fraction appears protective. The independent contribution of very-low density lipoprotein and its triglyceride or cholesterol content has, on the other hand, not been established. The previous position that virtually all of the lipid information pertaining to coronary heart disease resided in the serum total cholesterol must be accordingly modified.


Opiate receptors: some anatomical and physiological aspects.

It is possible to localize opiate receptors by light microscopic autoradiography using techniques developed for the autoradiographic localization of small diffusible molecules. The receptors are found in the neuropil of anatomical areas associated with various physiologic functions known to be altered by opiates. In some areas, opiate receptors are associated with the axons of small diameter fibers. Many kinds of studies, especially electrophysiologic studies, provide evidence that receptors revealed in these studies have a physiological function. Electrophysiological and immunohistochemical studies of the enkephalins provide results which are compatible with the notion that the enkephalins are the endogenous substrates of the opiate receptors.


Pretracheal cervical granular cell myoblastoma.

This case report deals with a solitary granular cell myoblastoma extrinsic to the proximal trachea. This lesion can be encountered frequently by the plastic surgeon; its histology, incidence and treatment are reviewed here. It is important to reemphasize that local extirpation is appropriate treatment of what is almost always a benign neoplasm, and intraoperative pathological diagnosis guides the surgeon in providing curative therapy.


Juvenile amaurotic idiocy (neuronal ceroid lipofuscinosis) and lymphocyte fingerprint profiles.

Lymphocytes from 3 children with a form of juvenile amaurotic idiocy (characterized by retinal blindness, progressive dementia, and extrapyramidal motor disturbance) were studied by electron microscopy. Numerous fingerprint profiles (FP) were found in vacuolated lymphocytes from all 3 patients and an asymptomatic younger sibling of 1 patient who subsequently became symptomatic. We propose that the combination of this clinical picture and vacuolated lymphocytes with FP is sufficiently distinctive for clinical and research purposes until the biochemical defect is discovered. Wider utilization of ultrastructural study of lymphocytes should increase the number of children diagnosed and allow detection of asymptomatic patients.


Neurological manifestations of Fabry disease in female carriers.

A family is described in which a 33-year-old man has classic X-linked recessive Fabry disease. His 2 sisters were discovered to be heterozygous carriers of the Fabry gene and to have both episodic and permanent neurological deficits including vertigo, tinnitus, long tract motor signs, and bladder incontinence. The most concise explanation for these findings is that the sisters manifest central nervous system complications of the Fabry carrier state. This family provides additional evidence that female carriers of rare X-linked recessive disorders may exhibit serious consequences of the disease, presumably related to tissue variability in expression of mutant enzyme activity.


Treatment of experimental herpesvirus infections with phosphonoformate and some comparisons with phosphonoacetate.

Phosphonoformate (PF) at a concentration of 5 to 10 mug/ml inhibited the growth of type 1 strains of herpes simplex virus (HSV) in tissue culture, whereas 20 to 30 mug/ml was required for inhibition of type 2 strains and about 50 mug/ml was required for murine cytomegalovirus. In mice inoculated intraperitoneally or intracerebrally with HSV or intraperitoneally with murine cytomegalovirus, treatment with 250 to 400 mg of PF per kg twice daily for 5 days had only minimal effectiveness. When mice were inoculated intravaginally (i.vg.) with HSV type 2 and treated i.vg. with 10% PF beginning 3 h after viral inoculation, treatment was effective in completely inhibiting viral replication in the genital tract. If i.vg. therapy was initiated 24 h after infection, when the mice had a mean virus titer of 10(5) plaque-forming units in vaginal secretions, a significant reduction in the mean virus titer was observed on days 3, 5, and 7 after infection as compared with control animals. In guinea pigs treated i.vg. with 10% PF beginning 6 h after i.vg. inoculation with HSV type 2 there was also complete inhibition of viral replication in the genital tract, and no extenal lesions developed. When therapy was initiated 24 h after infection there was a 4 to 5-log decrease in viral titers on days 3, 5, and 7 of the infection and a slight delay in the development of external lesions.


Efficacy of sodium diethyldithiocarbamate (dithiocarb) in acute nickel carbonyl poisoning.

A brief historical resume is presented on the use of dithiocarbamates for the treatment of persons exposed to nickel carbonyl. The specificity of the treatment is demonstrated in an industrial accident in which four men were simultaneously exposed to nickel carbonyl vapors. Three of the men received dithiocarb within 24 hours after exposure and the fourth was hospitalized by his family physician and treated for bronchopneumonia with antibiotics and without benefit of dithiocarb. The three workmen who received dithiocarb became symptomless and returned to work within 72 hours after exposure. The fourth man who had not received dithiocarb died within five days after exposure.


Effect of 9-(2-hydroxyethoxymethyl)guanine on herpesvirus-induced keratitis and iritis in rabbits.

Drugs used for the inhibition of DNA viruses, such as iododeoxyuridine, adenine arabinoside, or trifluorothymidine, are not biochemically selective in their action and also interfere with normal cellular functions. The recently reported 9-(2-hydroxyethoxymethyl)guanine (acycloguanosine) is selectively phosphorylated by viral thymidine kinase but not by normal cellular thymidine kinase. Our present studies show that the acycloguanosine is as effective in treating herpetic keratitis in the rabbit as iododeoxyuridine and trifluorothymidine when given topically as an ointment. It is also effective when given intravenously for the treatment of herpetic iritis and is effective in preventing death from encephalitis in rabbits.


Comparative study of ten bacteriocins of Clostridium perfringens.

Bacteriocins of Clostridium perfringens were prepared by ammonium sulfate precipitation of supernatant broth from 10 bacteriocinogenic strains. These bacteriocins were compared with respect to their ability to produce spheroplasts in a sensitive indicator strain; their inducibility; sensitivity to pH, proteolytic enzymes, and boiling; and their effect on macromolecular synthesis. Two bacteriocins were stable over a wide range of pH values and resisted boiling, and three bacteriocins were resistant to trypsin. Five bacteriocins shut down DNA, RNA, and protein synthesis; three bacteriocins had varying effects on DNA and RNA synthesis; and two bacteriocins had little effect on macromolecular synthesis. Antiserum prepared against one bacteriocin highly neutralized three bacteriocins with partial neutralization of five others; two bacteriocins were unaffected. Mutant strains selected for resistance to bacteriocin 28 also demonstrated coresistance to two other closely related bacteriocins and partial resistance to five others.


Polymyxin B-penicillin antagonism in Proteus mirabilis.

Polymyxin B protects Proteus mirabilis from usually lethal penicillin concentrations.


Cell killing by Simian virus 40: protective effect of chloroquine.

Treatment of CV-1 cells with chloroquine before infection by simian virus 40 resulted in the accumulation of fewer nonviable, trypan blue-stainable cells at 72 h. The drug did not affect the fraction of infected T-antigen-producing cells or the viral yields. It did diminish the apparent redistribution of lysosomal N-acetyl-beta-glucosaminidase from a particulate to a soluble cell fraction, and it caused an increase in the size and number of lysosomes.


On betaH-Leu5-endorphin and schizophrenia.

A previously unknown peptide, betaH-Leu5-endorphin, has been reported in the dialysates of schizophrenic patients. Accordingly, hemofiltrates from two schizophrenic and two control patients were examined for the presence of betaH-Leu5-endorphin. The opioid peptides were detected by a radioreceptor assay after separation and identification by gel filtration and high-performance liquid chromatography. With a detection limit of 30 pmole/L of hemofiltrate, no betaH-Leu5-endorphin or Met5-endorphin was found in controls or in patients. Whatever the possible involvement of endorphins in schizophrenic behavior, they are not present at detectable levels in the hemofiltrates of two well-characterized schizophrenic patients, thereby casting doubt on a general relationship of Leu-endorphin and schizophrenia.


[Pancreatic lesion in newborn mice caused by Coxsackie B1 virus].

Changes in the pancreatic gland manifested by necrosis of excretory cells and marked atrophy of insulae with the loss of beta-cells producing insulin were observed experimentally in suckling mice infected with Coxsackie B1 virus. It is suggested that the above changes of the pancreas may be a model of human diseases caused by Coxsackie viruses, including a model of development of some cases of diabetes.


'Locked-in coma' in postinfective polyneuropathy.

A patient with postinfective cranial and peripheral polyneuropathy exhibited the electroencephalographic and behavioral features of "alpha coma". The relation of this form of extensive peripheral disconnection to those cases with central disconnection due to pontomesencephalic lesions is discussed. We conclude that in both situations further evaluation of brain stem and cortical function is necessary to determine whether or not consciousness is preserved, rather than relying solely on the presence of ocular movements and reactivity of the electroencephalogram.


Russell bodies in contact-lens-associated giant papillary conjunctivitis.

Biopsy specimens of the upper tarsal conjunctiva in soft contact lens-associated giant papillary conjunctivitis were taken during (1) chronic exacerbation, (2) brief remission, and (3) intentional exacerbation. Inflammatory cells were quantitated and compared with inflammatory cells in normal upper tarsal conjunctivae. Specimens were evaluated by light and electron microscopy. The most remarkable feature was the presence of diamond-shaped Russell bodies in 20% of the plasma cells of the second biopsy specimen. A few round Russell bodies were seen in the first biopsy specimen and none in the third. We concluded that the brief quiescent phase (second biopsy specimen) was characterized by retention of immunoglobulin to produce Russell bodies, and that the active phases of the disease were marked by migration of mast cells into the epithelium and by the presence of eosinophils and basophils in the substantia propria.


Psoriasis: a medical mystery?

Psoriasis remains a disease of unknown aetiology. It is common, has a chronic course with exacerbations and remissions, and may present at any age. There are a number of clinical types recognized and an association with arthritis has been noted. Treatment is directed at control of the disease to enable the person to live and cope with his disease state. This can often be achieved with topical therapy, but it may at times require hospitalization. More severe forms of psoriasis may require potentially dangerous medication.


Primary, severe, combined immunodeficiency disease of Arabian foals.

Set in a context of immunodeficiency diseases in general this paper provides a brief, illustrated review of a primary, severe, combined immunodeficiency (PSCID) disease of Arabian foals. Affected foals are clinically normal at birth but beginning at about 10 days of age they develop a range of clinical signs particularly bronchopneumonia and diarrhoea with which adenoviruses are peculiarly associated. Despite intensive therapy foals invariably die by about 3 months of age. Affected foals are profoundly lymphopagenic (greater than 1000 lymphcoytes per mm3). There is thymic and lymph node hypoplasia and all lymphoid tissues are profoundly depleted of both T and B lymphocytes. The depletion of both T and B lymphocytes suggests that the primary defect is at the level of bone marrow stem cells which are the precursor cells for both lymphocyte populations. PSCID of Arabian foals is inherited as a simple, autosomal, recessive gene. Some 2 to 3% of all such foals may be born with PSCID, this frequency corresponds to a gene frequency of about 30% in parents. The syndrome is, therefore, an important cause of economic wastage. It also represents the only occurrence of the syndrome in an animal species other than man and as such has considerable comparative interest.


The hypercoagulable state and pulmonary embolism in patients with ovarian carcinoma.

Fifty-nine patients with ovarian carcinoma were studied with particular reference to abnormalities of the coagulation mechanism. The notable findings were the high incidence of episodic thrombocytosis (73%) and the relation of these peaks of thrombocytosis to periods of expected maximum tumour break-down. The incidence of pulmonary embolism for the series was 44%, with the majority occurring in the group demonstrating significant thrombocytosis; in this latter group, pulmonary embolism constituted the major cause of death. The abnormalities in coagulation are discussed, together with pragmatic significance and methods of control.


Application of a bubble formation model to decompression sickness in rats and humans.

Although decompression sickness results from bubble formation in blood or tissue, pressure schedules currently in use are essentially empirical and contain little input from cavitation theory. The recent convergence of three lines of investigation suggests that a synthesis of practice and theory may now be possible. The data consist of pressure reduction limits for gelatin, rats, and humans following steady-state exposures. From the gelatin studies, a model has been developed in which bubble formation is initiated by spherical gas nuclei stabilized by surface-active skins of varying gas permeability. We demonstrate that the model is also in good agreement with data on rats and humans over a wide range of pressures and that the model parameters assume sensible values in each case. This suggests that cavitation theory can provide a rationale for current diving practice and can serve to secure, consolidate, and extend this practice.


Electrophoretic heterogeneity of 5-phosphoribosyl-1-pyrophosphate synthetase within and among humans.

The product of the enzyme 5-phosphoribosyl-1-pyrophosphate (PPriboseP) synthetase is a substrate for purine, pyrimidine, and pyridine biosynthesis and may be rate limiting for purine biosynthesis. A system developed to electrophoretically separate and histochemically detect PPriboseP synthetase in crude cell extracts has facilitated the identification of electrophoretically variant enzyme forms in the erythrocytes of five patients from a patient population of 200. Additional studies of human organs obtained at autopsy revealed a unique electrophoretic mobility for nearly each organ within the same individual.


Microheterogeneity of adenosine cyclic monophosphate-dependent protein kinases from mouse brain and heart.

1. DEAE-cellulose chromatography of mouse brain cytosol indicated the presence of only the type II isoenzyme of cyclic AMP-dependent protein kinase. Mouse heart cytosol contained approximately equal amounts of the type I and type II isoenzymes. 2. Both brain and heart type II isoenzymes reassociated after a transient exposure to cyclic AMP, but the heart type I isoenzyme remained dissociated. 3. Elution of brain cytosol continuously exposed to cyclic AMP resolved multiple peaks of protein kinase and cyclic AMP-binding activities. A single peak of kinase and multiple peaks of cyclic AMP-binding activities were found under the same conditions with heart cytosol. Various control experiments suggested that the heterogeneity within the brain type II isoenzymic class had not been caused by proteolysis. 4. Kinetic experiments with unfractionated brain cytosol showed that the binding of cyclic AMP, the dissociation of cyclic AMP from protein and the rate of heat denaturation of the cyclic AMP-binding activity gave results consistent with the presence of multiple binding species. 5. It concluded that the type II isoenzymic peak obtained by DEAE-cellulose chromatography of mouse brain cytosol represents a class of enzymes containing multiple regulatory and catalytic subunits. The two heart cytosol isoenzymes contain a common catalytic subunit. The degree of protein kinase 'microheterogeneity", defined as the presence of multiple regulatory and/or catalytic subunits within a single isoenzymic class, appears to be tissue-specific.


The metabolic modification of low-density lipoproteins in normal and hypercholesterolaemic guinea pigs.

1. Low-density lipoproteins were isolated by ultracentrifugation from the serum of guinea pigs that were fed either on a normal diet, or on a diet supplemented with corn oil and cholesterol. 2. After labelling with tracer amounts of radioactive iodine, these lipoproteins were injected into the bloodstream of guinea pigs that were fed either on the normal or on the supplemented diet. 3. In all cases, the density of the labelled lipoproteins was increased by exposure for 24-48 h to the metabolic processes of the guinea pig. 4. The final density reached by lipoproteins isolated from fat-fed guinea pigs was less than that reached by lipoproteins from normal animals. 5. Fat-fed guinea pigs were unable to increase the density of either normal lipoproteins, or those from fat-fed guinea pits, to the same extent as animals fed on the normal diet. 6. It is concluded that the lipid-rich diet brings about a modification of lipoprotein metabolism in the guinea, pig, which plays an important part in determining the nature of the nature of the low-density lipoprotein that is present in the plasma.


Effect of beta-lapachone on superoxide anion and hydrogen peroxide production in Trypanosoma cruzi.

Addition of beta-lapachone, an o-naphthoquinone endowed with trypanocidal properties to respiring Trypanosoma cruzi epimastigotes induced the release of O2- and H2O2 from the whole cells to the suspending medium. The same beta-lapachone concentration (4 micron) that released H2O2 at maximal rate completely inhibited T. cruzi growth in a liquid medium. The position  isomer, alpha-lapachone, did not stimulate O2- and H2O2 release, and did not inhibit epimastigote growth. beta-Lapachone was able to stimulate H2O2 production by the epimastigote homogenate in the presence of NADH as reductant. The same effect was observed with the mitochondrial fraction supplemented with NADH, where beta-lapachone enhanced the generation of O2- and H2O2 4.5- and 2.5-fold  respectively. beta-Lapachone also increased O2- and H2O2 production (2.5 and 2-fold respectively) by the microsomal fraction with NADPH as reductant. Cyanide-insensitive NADH and NADPH oxidation by the mitochondrial and microsomal fractions (quinone reductase activity) was stimulated to about the same extent by beta-lapachone. alpha-Lapachone was unable to increase O2- and H2O2 production and quinone reductase activity of the mitochondrial and microsomal fractions.


Incorportion of oxygen-18 into the 25-position of cholecalciferol by hepatic cholecalciferol 25-hydroxylase.

The oxygen enzymically inserted as a hydroxy function by rat liver post-mitochondrial fraction into the 25-position of cholecalciferol to giver 25-hydroxycholecaliferol is derived exclusively from molecular O2. Therefore like the other two cholecalciferol hydroxylases, i.e. 25-hydroxycholecalciferol 1alpha-hydroxylase and 25-hydroxycholecalciferol 24-hydroxylase, the cholecalciferol 25-hydroxylase is also a mono-oxygenase ('mixed-function oxidase').


Purification and properties of nitrite reductase from Escherichia coli K12.

NADH-nitrite oxidoreductase (EC 1.6.4) was purified to better than 95% homogeneity from batch cultures of Escherichia coli strain OR75Ch15, which is partially constitutive for nitrite reductase synthesis. Yields of purified enzyme were low, mainly because of a large loss of activity during chromatography on DEAE-cellulose. The quantitative separation of cytochrome c-552 from nitrite reductase activity resulted in an increase in the specific activity of the enzyme: this cytochrome is not therefore an integral part of nitrite reductase. The subunit molecular weights of nitrite reductase and of a haemoprotein contaminant, as determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, were 88000 and 80000 respectively. The sedimentation coefficient was calculated to be in the range 8.5-9.5S, consistent with a mol.wt. of 190000. It is suggested therefore that the native enzyme is a dimer with two identical or similar-sized subunits. Purest samples contained 0.4 mol of flavin/mol of enzyme, but no detectable haem. Catalytic activity was totally inhibited by 20 micron-p-chloromercuribenzoate and 1 mM-cyanide, slightly inhibited by 1 micron-sulphite and 10mM-arsenite, but insensitive to 1 mM-2,2'-bipyridine, 4mM-1,10-phenanthroline and 10mM-NaN3. Three molecules of NADH were oxidized for each NO2-ion reduced: the product of the reaction is therefore assumed to be NH4+. The specific activity of hydroxylamine reductase increased at each step in the purification of nitrite reductase, and the elution profiles for these two activities during chromatography on DEAE-Sephadex were coincident. It is likely that a single enzyme is responsible for both activities.


Activation of nitrite reductase from Escherichia coli K12 by oxidized nicotinamide-adenine dinucleotide.

Nitrite reductase from Escherichia coli K12 requires the presence of NAD+, one of the products of the reduction of NO2-by NADH, for full activity. The effect is observed with both crude extracts and purified enzyme. NAD+ also acts as a product inhibitor at high concentrations, and plots of initial rate against NAD+ concentration are bell-shaped. The maximum occurs at about 1 mM-NAD+, but increases with increasing NADH concentration. In the presence of 1 mM-NAD+ and saturating NO2-(2mM) the Michaelis constant for NADH is about 16 micron. The Michaelis constant for NO2-is about 5 micron and is largely independent of the NAD+ concentration. Similar but more pronounced effects of NAD+ are observed with hydroxylamine as electron acceptor instead of NO2-. The maximum rate of NADH oxidation by hydroxylamine is about 5.4 times greater than the maximum rate of NADH oxidation by NO2- when assayed with the same volume of the same preparation of purified enzyme. The Michaelis constant for hydroxylamine is 5.3 mM, however, about 1000 times higher than for NO2-. These results are consistent with a mechanism in which the same enzyme-hydroxylamine complex occurs as an intermediate in both reactions.


The specific interaction of cibacron and related dyes with cyclic nucleotide phosphodiesterase and lactate dehydrogenase.

1. Reactive Blue 2 (Cibacron Blue 3G-A) is a competitive inhibitor of bovine heart cyclic nucleotide phosphodiesterase (K(i) 0.3mum). The K(i) increases with increasing temperature, suggesting that hydrophobic interactions are not largely responsible for the binding of the dye. Another 25 sulphonated aromatic dyes are also competitive inhibitors of the cyclic nucleotide phosphodiesterase (K(i) values in the range of 0.06-13.6mum). 2. These dyes (covalently linked to Dextran 40) inhibit bovine heart cyclic nucleotide phosphodiesterase. Reactive Blue 2 (covalently linked to Dextran 40) is a competitive inhibitor of the phosphodiesterase (K(i) 0.4mum). 3. Bovine heart cyclic nucleotide phosphodiesterase is retained on a column of Reactive Blue 2-Sephacryl S-200 and can be eluted from the column by 3':5'-cyclic AMP. 4. A variety of the dyes (either free or covalently linked to Dextran 40) are competitive inhibitors of rabbit muscle lactate dehydrogenase. 5. The effectiveness of a wide range of structurally dissimilar dyes as competitive inhibitors of lactate dehydrogenase and cyclic nucleotide phosphodiesterase compromises proposals for the use of Reactive Blue 2 as a specific probe for the dinucleotide-binding structural domain present in many dehydrogenases and kinases. Detailed information of the various dyes used has been deposited as Supplementary Publication SUP 50089 (7 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1978) 169, 5.


Inhibition of deoxyribonucleic acid polymerases from human cells and from simian sarcoma virus by pyran.

Pyrans are co-polymers of divinyl ether and maleic anhydride. Four pyrans of various molecular weights more potently inhibited terminal deoxyribonucleotidyltransferase (EC 2.7.7.31) from a human cell line of acute lymphoblastic leukemia origin (Molt-4) than they did DNA polymerases alpha, beta and gamma from these cells and DNA polymerase from simian sarcoma virus. For example, the concentrations of one pyran required for 50% inhibition of terminal deoxynucleotidyltransferase, DNA polymerases alpha, beta and gamma and viral DNA polymerase were 0.9, 110, 125, 35 and 47 microgram/ml respectively. Quantitatively similar results were obtained with the other pyrans. Inhibition of these enzymes by pyran was dependent on the concentrations of both the bivalent cation and template/primer or initiator in assay mixtures, but not on the concentrations of the substrate (deoxyribonucleoside 5'-triphosphate), enzyme, or bovine serum albumin. These results suggested that pyran inhibited these enzymes by complexing bivalent cations, which caused a decreased affinity of template/primer or initiator for each enzyme and a decrease in enzyme activity.


Oxidation of 2-nitropropane by horseradish peroxidase. Involvement of hydrogen peroxide and of superoxide in the reaction mechanism.

Incubation of aqueous solutions of 2-nitropropane in air causes a slow oxidation reaction that generates H(2)O(2). Purified horseradish peroxidase catalyses the oxidation of such preincubated 2-nitropropane solutions according to the equation: [Formula: see text] The pH optimum is 4.5 and K(m) for 2-nitropropane is 16mm. Other nitroalkanes or nitro-aromatics tested are not oxidized at significant rates by peroxidase. H(2)O(2) or 2,4-dichlorophenol increases the rate of 2-nitropropane oxidation by peroxidase. Catalase inhibits the reaction completely. Superoxide dismutase or mannitol, a scavenger of the hydroxyl radical, OH(.), each inhibits partially. Aniline and guaiacol are also powerful inhibitors of 2-nitropropane oxidation. It is suggested that peroxidase uses the traces of H(2)O(2) generated during preincubation of 2-nitropropane to catalyse oxidation of this substrate into a radical species that can reduce O(2) to the superoxide ion, O(2) (-.).O(2) (-.), or OH(.) derived from it, then appears to react with more nitropropane, generating further radicals and H(2)O(2) to continue the oxidation. Inhibition by aniline and guaiacol seems to be due to a competition for H(2)O(2).


An enzyme degrading reduced nicotinamide-adenine dinucleotide in Proteus vulgaris.

Cell-free extracts of a strain of Proteus vulgaris degrade NADH to reduced nicotinamide riboside, adenosine and two molecules of phosphate. The system is weakly active in fresh cell extracts, but activity is increased about 10-fold on rapid heating to 70-100 degrees C. On returning to room temperature, the activity returns rapidly to its initial low value but can be re-activated by again heating to 70-100 degrees C. Reversible activation can also be effected by extremes of pH or by teatment with 8M-urea. Activation appears to be due to reversible changes in conformation of the protein of the enzyme rather than to combination of the enzyme with a heat-labile inhibitor. The active form can be stabilized by addition of PPi. The system, which also possesses 5'-nucleotidase activity not separable from the NADH pyrophosphatase, requires Co2+ (0.4mM) for maximum activity. Although activated at relatively high temperatures, it is not enzymically active until cooled to 50-60 degrees C. It may be purified by affinity chromatography (with NAD+ as ligand) to an activity over 400 times that of the crude cell extract, and yields only one major band on polyacrylamide-gel electrophoresis.


The kinetics and thermodynamics of the reaction of solid-state fully reduced membrane-bound cytochrome oxidase with carbon monoxide as studied by dual-wavelength multichannel spectroscopy and flash photolysis.

1. The results of non-linear optimization studies on the mechanism of reaction of solid-state fully reduced membrane-bound cytochrome oxidase with CO over the 178--203 K range are presented. The analysis is carried out on data obtained by dual-wavelength multichannel spectroscopy at three wavelength pairs (444--463 nm, 590--630 nm and 608--630 nm), which yield three distinct progress curves. The only model that satisfies the triple requirement of a standard deviation within the standard error of the data, a random distribution of residuals and good determination of the optimized parameters is a two-species sequential mechanism: flash photolysis yields unliganded cytochrome oxidase and free CO, which then recombine to form species Ic; Ic is then converted into species IIc, which is identical with the cytochrome oxidase-CO complex existing before flash photolysis. All the thermodynamic parameters describing this model are calculated. 2. On the basis of the data obtained from this paper, together with data from potentiometric studies, magnetic susceptibility measurements and i.r. spectroscopy, the chemical identity of the species is suggested.


Partial reversal of the acetaldehyde and butyraldehyde oxidation reactions catalysed by aldehyde dehydrogenases from sheep liver.

In the presence of acetic anhydride or butyric anhydride, liver aldehyde dehydrogenases catalyse the oxidation of NADH at pH 7.0 and 25 degrees C. The maximum velocities and Michaelis constants for NADH at saturating anhydride concentrations are independent of which anhydride is used, the values being V'max. = 12 min-1 and Km for NADH = 9 micrometer for the mitochondrial enzyme and V'max = 25 min-1 and Km for NADH = 20 micrometer for the cytoplasmic enzyme. Substitution of [4A-2H]NADH for NADH resulted in 2-fold and 4-fold decreases in rate for the mitochondrial and cytoplasmic enzymes respectively.


Stimulation of intestinal calcium-binding-protein mRNA synthesis in the nucleus of vitamin D-deficient chicks by 1,25-dihydroxycholecalciferol.

Stimulation of intestinal calcium transport by the hormone 1,25-dihydroxycholecalciferol appears to involve RNA transcriptions and the synthesis of new proteins. Although one of these proteins has been identified as calcium-binding protein, no RNA molecules specifically induced by the hormone in the nucleus have been identified. Nuclear RNA from intestine of vitamin D-deficient chicks before and at various time intervals after treatment with the hormone or cholecalciferol was tested for its ability to code for calcium-binding protein in a cell-free system. Calcium-binding-protein mRNA could only just be detected in the intestinal nuclei 2h after dosing with these steroids which is the same time that it was first observed in the polyribosomes. Thus 1,25-dihydroxycholecalciferol induces the production of new calcium-binding protein by stimulating the formation and rapid release from the nucleus of new mRNA molecules for this protein. Polyribosomal translation of the mRNA continued only as long as it was being synthesized, and the maximum rate of synthesis following a pulse dose of 125ng of the hormone was the same as that observed after prolonged stimulation with cholecalciferol. The possibility that other 1,25-dihydroxycholecalciferol-dependent events may be occurring in the nucleus in the lag period between accumulation of the hormone in the intestine and the appearance of active calcium-binding-protein mRNA, and that these may ultimately control the synthesis of that mRNA, is discussed.


The effect of measuring light photolysis on the kinetics of reaction of reduced cytochrome a3 and carbon monoxide.

The kinetics of reaction of reduced cytochrome a3 and CO are re-investigated by non-linear optimization techniques. When photolysis by the monitoring light is taken into account, the experimental data are best fitted by a single-intermediate mechanism.


A new carbon monoxide-induced complex of cytochrome c oxidase.

Cytochrome c oxidase isolated from ox heart forms a complex in the presence of millimolar concentrations of CO with absorption bands at 606, 565 and 435 nm (difference spectrum), distinct from both ferrocytochrome a and the classical 590nm carbon-monoxyferrocytochrome a3. This species, which closely resembles Compound C, the derivative formed on photolysis and oxygenation of mixed-valence cytochrome a3+a32+CO, may represent a cytochrome a32+CO complex in which the associated ('invisible') copper is still oxidized.


The molybdenum centre of native xanthine oxidase. Evidence for proton transfer from substrates to the centre and for existence of an anion-binding site.

The observation by Bray & Knowles [Proc. R. Soc. London Ser. A (1968) 302, 351--353] of direct transfer, during the catalytic reaction, of hydrogen atoms from substrate molecules to the enzyme xanthine oxidase was reinvestigated. The experimental phenomenon and its basic interpretation were confirmed and extended. In the reduced functional enzyme, molybdenum(V) interacts with two enzyme-bound protons, which are exchangeable with solvent protons. One of these is coupled to the metal with AHav. 1.4mT and the other with AHav. 0.3mT. The molecule also contains a site for the binding of anions, presumably as ligands of molybdenum. This is shown by effects of nitrate ions on the e.p.r. spectra. The spectra of the nitrate and 1-methylxanthine complexes of the reduced enzyme are very similar to one another, and are designated Rapid type-1 spectra. It is concluded that, in the Michaelis complex, the substrate molecule occupies the anion site, probably being bound to molybdenum via the nitrogen in its 9-position. During the turnover process, hydrogen from the substrate C-8 position, after transfer to the enzyme, appears as the proton more strongly coupled to molybdenum. This proton then exchanges with solvent deuterium with a rate constant of 27s-1, at pH 8.2 and 12 degrees C. It has been confirmed that substrate molecules occupying the anion site do not interfere with observation of the transfer and exchange processes.