Cholinergic agents inhibit sodium transport across the isolated toad bladder.

Acetylcholine and carbamylcholine (carbachol) inhibited sodium transport across the toad bladder. This effect was blocked by atropine. Carbachol increased the uptake of 45Ca by isolated toad bladder epithelial cells. This increased 45Ca uptake was blocked by atropine, pentobarbital, or lanthanum chloride. The inhibitory effect of cholinergic agents on sodium transport was dependent on external calcium concentration and was abolished by decreasing external calcium from 2 to 0.2 mM or by agents (pentobarbital, lanthanum chloride) which prevent the movement of calcium into cells. Carbachol increased modestly, but significantly, the levels of cyclic GMP in isolated toad bladder epithelial cells. This effect was blocked by atropine. Whether the decrease in sodium transport produced by cholinergic agents is the result of increased intracytoplasmic calcium levels, increased cyclic GMP levels, or a combination of both remains to be established.


Long-term effects of traumatic war-related events on sleep.

Eleven patients who had combat neuroses resulting from the 1973 Yom Kippur War and complained of sleep disturbances were studied in a sleep laboratory. Sleep-onset insomniacs, dream-interruption insomniacs, and pseudoinsomniacs were differentiated on the basis of electrophysiologic recordings. Compared with normal controls who actively participated in the Yom Kippur War, patients showed significantly longer sleep latencies, lower sleep efficiency indices, lower percentage of REM sleep, and longer REM latencies.


Psychiatric services in Soviet general outpatient clinics.

The author believes that the first level of psychiatric care in the U.S.S.R.--the general polyclinics (outpatient clinics) and the medical-sanitary units--have not been adequately studied. He describes this level of care, which is considered useful and progressive in that the clinics are supposed to make psychiatric care available to the population, lighten the load placed on psychiatric facilities, and improve follow-up therapy. However, he also points out the shortcomings in this first level of psychiatric care in the U.S.S.R., including the bureaucratic nature of the system and the uneven quality of service the clinics render to various social groups in urban and rural areas.


Malakoplakia. Two unusual cases which presented diagnostic problems.

Two unusual cases of malakoplakia are presented. Both cases occurred outside the urinary tract (retroperitoneum and anus) and were diagnostic problems due to the paucity of Michaelis-Guttman bodies. One case mimicked a retroperitoneal malignant fibrous histiocytoma and the other a common perianal abcess.


Rhabdomyosarcoma arising within a cystosarcoma phyllodes. Case report and review of the literature.

Rhabdomyosarcoma of the breast is a rare tumor that is characterized by its rapid growth, large size, and poor prognosis. It most often presents as a pure neoplasm and infrequently as the stromal component of a cystosarcoma phyllodes. The clinical and pathological features of a cystosarcoma with a rhabdomyosarcomatous stroma occurring in a 45-year-old woman are presented, and the results are discussed in reference to the 24 published cases of mammary rhabdomyosarcoma. The patient was treated by radical mastectomy and died 2.5 years later with pulmonary and cerebral metastases. Though two long-term survivors have been reported, cures are infrequent and no form of therapy has been uniformly successful.


Radioactive and nonradioactive methods for the in vivo determination of disulfiram, diethyldithiocarbamate, and diethyldithiocarbamate-methyl ester.

Although disulfiram (tetraethylthiuram disulfide; DSF) has been used in the treatment of alcoholism for almost a quarter of a century, little is known about its in vivo metabolism. One reason for this is that few analytical methods are available that can determine DSF and its various metabolites in biologic fluids and tissues. This article describes two simple procedures for the determination of these substances.


Methods for detecting disulfiram in biologic fluids: application in studies of compliance and effect of divalent cations on bioavailability.

Studies regarding the efficacy of disulfiram in the treatment of alcoholism have not included a method of evaluating compliance to the prescribed regimen. This article outlines current methods used to detect disulfiram in blood, breath, and urine.


Characterization of Sertoli cell-germ cell junctional specializations in dissociated testicular cells.

To further characterize Sertoli cell-germ cell junctional specializations seminiferous tubules from sexually mature Sprague-Dawley rats were dissociated by enzymatic and mechanical methods. Ultrastructural analysis of cell suspensions prepared by incubation in collagenase alone or by mechanical methods revealed that spermatids remained attached to Sertoli cells or Sertoli cell fragments. Such cellular associations were found only between Sertoli cell fragments and spematids in which the developing acrosome had made contact with the plasma membrane (step 8 and subsequent steps of spermiogenesis). Furthermore, the fragments were confined to that region of the plasma membrane over the acrosome. The Sertoli cell half of this adhesive site displayed the typical elements of Sertoli cell junctions, filamentous bundles and associated cisterna of endoplasmic reticulum, in apposition to the spermatids. The spermatids demonstrated no surface specializations at the attachment sites. In contrast, in cell suspensions prepared with trypsin, spermatids were free of attachments to Sertoli cells or their fragments. These results demonstrate that: (1) the junctions act to bind cells together, (2) adhesive type contact is established between Sertoli cells and spermatids at step 8 and subsequent steps of spermiogenesis, (3) contact is restricted to the spermatid plasma membrane over the acrosome, and (4) spermatids can be freed from the junctional specializations by treatment with trypsin.


Ultracytochemical localization of Na+,K+-activated ATPase in chloride cells from the gills of a euryhaline teleost.

The activity of the electrolyte transport enzyme, sodium, potassium-activated adenosine triphosphatase (Na+,K+-ATPase), in the gills of the pinfish, Lagodon rhomboides, increased markedly following transfer of fish from brackish water to seawater. Cytochemical localization of Na+,K+-ATPase via its potassium-dependent phosphatase (K+-NPPase) activity in the branchial epithelium of pinfish adapted to seawater demonstrated that chloride cells are the major sites for the enzyme. Subcellularly, the heaviest depositions of reaction product were observed lining the cytoplasmic membrane surfaces of the labyrinth of anastomosing plasma membrane tubules that ramifies throughout the chloride cell cytoplasm. Enzyme activity was demonstrated also on the cytoplasmic surface of the apical crypt membrane and on the cytoplasmic surfaces of vesicles in the cytoplasm subjacent to the crypt. Deletion of potassium from the cytochemical incubation medium or inclusion of 10 mM ouabain abolished the reaction products associated with these membranes. The significance of these cytochemical results is discussed with reference to current hypotheses of chloride cell function.


Effects of Haemophilus influenzae vaccination on the (para-)sympathic-cyclic nucleotide-histamine axis in rats.

To determine whether Haemophilus influenzae could be a factor in human atopy its effects were studied on the (para-)Sympathic Cyclic nucleotide-histamine axis in rats. Haemophilus influenzae vaccination induced changes in the cholinergic system compatible with higher cyclic GMP levels and enhanced histamine release. The authors suggest an involvement of the cholinergic system in Haemophilus influenzae vaccination effects.


[Antigen enzyme coupling: a few results (author's transl)].

Various technics of coupling (carbodiimide, glutaraldehyde, periodate) have been used for the grafting of human casein on glucose oxidase. The various types of product obtained were identified by gel filtration and used for drawing up titration curves. The authors compared radioimmunoassay and enzymoimmunologic estimation showing the importance of the molecular size of the conjugates obtained.


Canine plasma cortisol (hydrocortisone) measured by radioimmunoassay: clinical absence of diurnal variation and results of ACTH stimulation and dexamethasone suppression tests.

A radioimmunoassay for plasma cortisol (hydrocortisone) was developed and validated for sensitivity, specificity, accuracy, precision, and parallelism. Steroids were extracted with ethyl ether, and cortisol was purified by gel column chromatography prior to assay. [1,2-3H] cortisol and a commercially available sheep antibody to cortisol-21-hemisuccinate were used. Free steriods were separated from bound steroids by centrifugation after adsorption to dextran-coated charcoal. Plasma cortisol was measured by this technique in 6 normal dogs. Circadian rhythm of cortisol secretion was not detected in samples obtained by venipuncture at 8 different hours on 3 separate days, suggesting that adrenal function tests may be started in clinical patients at any time of day. Resting plasma cortisol concentrations averaged 19.4+/-3.0 (SD) ng/ml and ranged from nondetectable (less than 3 ng/ml) to 77.5 ng/ml. Of 144 canine plasma samples, 95% contained less than 50 ng of cortisol/ml. Intramuscular injection of 2.2 units of adrenocorticotropic hormone/kg of body weight caused detectable increase in plasma cortisol concentrations; maximum response (68.3 to 111.6 ng/ml) occurred 1 to 2 hours after injection. Oral administration of dexamethasone suppressed plasma cortisol to nondetectable concentrations for 32 hours in all 6 dogs.


Simple method for preparation of specific antisera against viral proteins: rabbit antisera against equine infectious anemia virus proteins p26 and p16.

A simple method for preparation of highly specific antisera against equine infectious anemia virus proteins p26 and p16 is described. Viral proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The motility of the viral proteins in the gel was compared with standards. Unstained portions of the slab gel were sliced into 5-mm bands, emulsified with Freund's complete adjuvant, and injected into rabbits to produce specific antisera.


Recurrent hemorrhage in traumatic hyphema.

A retrospective review of hospital records reveals that over 90% of recurrent bleeds in traumatic hyphema occur at night. It is postulated that rapid eye movement (REM) sleep, with its displays of saccadic virtuosity, is the major reason for the nocturnal prevalence of such recurrent hyphemas. Theoretical considerations suggesting a more rational therapy of traumatic hyphema are presented and clinical trials are suggested.


Serum and CSF antibody levels to herpes simplex type 1, measles and rubella viruses in patients with schizophrenia.

Serum and CSF specimens from 12 schizophrenic patients and 10 non-psychiatric controls were tested for herpes simplex type 1 virus neutralizing antibody and for measles and rubella haemagglutination inhibiting antibodies. There were no significant differences in the distribution of virus antibody titres in serum or CSF specimens between the patients and the controls. The possible aetiological role of viruses or virus-like agents in schizophrenia and some methodological aspects are discussed.


[Blue rubber bleb nevus (author's transl)].

Hereditary dysembryoplasia, the B. R. B. N. (Bean, 1958) is a variety of bluish nevus of the skin, associated with angioma of the gastrointestinal tract with serious bleeding, and sometimes of liver, spleen, bones, brain, marrow. The angiomas of the skin may occur in 3 forms: large cavernous angiomas; blood sac looking like a blue rubber nipple, they can be emptied; irregular blue mark, sometimes with puncted blackish spots, they may not blanch on pressure. They are painful. glomus-like angioma cavernous angioma. Ultrastructure: cavity with endothelial cells and one to four coats of smooth muscle cells. The angiomas arose in the submucous cellular tissue, projected into the cavity of the bowel, sometimes bleeding. Hematemesis and melena may be observed as in the Rendu-Osler disease. They nver spontaneously involve but they don't degenerate. The prognosis depends essentially on the bleedings and anemia. It is possible to cut out the skin angiomas.


Chiasmal syndrome in sphenoid sinus mucocele.

Two patients with sphenoethmoid mucoceles developed visual field defects consistent with optic chiasm dysfunction. The presence of chiasmal field defects associated with the clinical syndrome of compressive optic neuropathy should not rule out the diagnosis of sphenoethmoid mucocele since definitive treatment can be carried out via transnasal drainage.


Endogenous pain control mechanisms: review and hypothesis.

The anatomy, physiology, and pharmacology of an intrinsic neural network that monitors and modulates the activity of pain-transmitting neurons is reviewed. This system can be activated by opiate administration or by electrical stimulation of discrete brainstem sites. Evidence is presented that its pain-suppressing action is mediated in part by endogenous opiatelike compounds (endorphins). This pain suppression system is organized at three levels of the neuraxis: midbrain, medulla, and spinal cord. Activation of neurons in the midbrain periaqueductal gray matter (by electrical stimulation, opiates, and possibly psychological factors) excites neurons of the rostral medulla, some of which contain serotonin. The medullary neurons, in turn, project to and specifically inhibit the firing of trigeminal and spinal pain-transmission neurons. As part of a negative feedback loop, the output of the pain transmission neurons, i.e., pain itself, is an important factor in activating the pain-suppression system. A neural model which incorporates the experimental findings is proposed, and the clinical implications of the model are discussed.


[Study of the hypothalamo-pituitary adrenal function in 16 patients after surgery for pituitary tumor (author's transl)].

The hypothalamic pituitary adrenocortical function has been studied in 16 patients operated from pituitary tumors (13 adenomas; 3 craniopharyngiomas). Comparisons have been made between corticotropin and cortisol response to lysine vasopressin, insulin induced-hypoglycemia and metyrapone IV and per os. Among these different stimulating tests, insulin induced hypoglycemia and metyrapone per os seem to give the more accurate informations metyrapone per os being more convenient because harmless. Three different groups of patients have been distinguished : one without adrenocortical deficiency; one with a complete deficiency and a third group with a partial deficiency. Correlations have been studied between the degree of the adrenocortical deficiency, the volume of the tumor and the presence of the absence of other anterior pituitary dysfunctions.


Correlation between hemin content and the chemiluminescent luminol reaction with bacteria.

Protohemin and covalently bound hemin were determined in eight aerobic bacterial strains. A good correlation between protohemin content and luminol reactivity was found. The ratio of luminol reaction to protohemin for the eight investigated strains was essentially identical to that of pure protohemin, 0.7 X 10(16) mV/mol. Covalently bound hemin contributed to the chemiluminescence to a minor extent only (0.7 X 10(14) mV/mol, in accordance with earlier observations of the lower reactivity of cytochrome c and related compounds. A difference in reaction kinetics of the luminol reaction with covalently bound hemin (slower reaction than protohemin) and protohemin was observed in vivo as well as in vitro. The phenomenon could be used to differentiate between strains with different hemin composition.


High levels of microbial contamination of vegetables irrigated with wastewater by the drip method.

The public health aspects of the use of wastewater in agriculture and the effects of the drip irrigation method on the contamination of vegetables were studied. The method used was to simulate enteric microorganisms' dissemination by contaminated irrigation water in the field. The vegetables were irrigated with an effluent inoculated with a high titer of traceable microorganisms: poliovirus vaccine and a drug-resistant Escherichia coli. The dissemination of the marker organisms in the field was followed, and the effects of certain manipulations of the drip irrigation method on the contamination of the crops by the effluent were examined. It was shown that drip irrigation under plastic sheet cover with the drip lines placed either on the soil surface or buried at a depth of 10 cm significantly reduced crop contamination from inoculated irrigation water even when massive doses of bacteria and viruses were used. The microbial contamination was found to persist in the irrigation pipes and in the soil for at least 8 and 18 days, respectively. The data indicate that the recovery of the marker organisms was affected by soil texture and environmental conditions.


Growth and methanogenesis by Methanosarcina strain 227 on acetate and methanol.

Methanosarcina strain 227 exhibited exponential growth on sodium acetate in the absence of added H(2). Under these conditions, rates of methanogenesis were limited by concentrations of acetate below 0.05 M. One mole of methane was formed per mole of acetate consumed. Additional evidence from radioactive labeling studies indicated that sufficient energy for growth was obtained by the decarboxylation of acetate. Diauxic growth and sequential methanogenesis from methanol followed by acetate occurred in the presence of mixtures of methanol and acetate. Detailed studies showed that methanol-grown cells did not metabolize acetate in the presence of methanol, although acetate-grown cells did metabolize methanol and acetate simultaneously before shifting to methanol. Acetate catabolism appeared to be regulated in response to the presence of better metabolizable substrates such as methanol or H(2)-CO(2) by a mechanism resembling catabolite repression. Inhibition of methanogenesis from acetate by 2-bromoethanesulfonate, an analog of coenzyme M, was reversed by addition of coenzyme M. Labeling studies also showed that methanol may lie on the acetate pathway. These results suggested that methanogenesis from acetate, methanol, and H(2)-CO(2) may have some steps in common, as originally proposed by Barker. Studies with various inhibitors, together with molar growth yield data, suggest a role for electron transport mechanisms in energy metabolism during methanogenesis from methanol, acetate, and H(2)-CO(2).


Identification of detergents as components of wastewater sludge that modify the thermal stability of reovirus and enteroviruses.

The agent in wastewater sludge previously shown to reduce the heat required to inactivate reovirus (R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol. 34:681--688, 1977) was "separated" from other sludge components and analyzed by infrared spectroscopy. The infrared spectrum of this material was quite similar to the spectra of commercial anionic detergents, and subsequent analyses of the fractionated sludge samples revealed that anionic detergents in sludge were copurified with the virucidal activity. Further measurements on the virucidal activities of specific detergents revealed that ionic detergents reduce the heat required to inactivate reovirus, that cationic detergents are more active than anionic, and that nonionic detergents are inactive. Several detergents were also shown to protect poliovirus and other enteroviruses against inactivation by heat. These results indicate that ionic detergents are the major component in wastewater sludge that reduce the thermal stability of reovirus and, in addition, that detergents are able to protect enteroviruses against heat.


Heat inactivation of enteric viruses in dewatered wastewater sludge.

The effect of moisture content on the rates of heat inactivation of enteric viruses in wastewater sludge was determined. The protective effect of raw sludge on poliovirus previously observed (R. L. Ward, C. S. Ashley, and R. H. Moseley, Appl. Environ. Microbiol. 32:339--346, 1976) was found to be greatly enhanced in sludge dewatered by evaporation. Other enteroviruses responded in a similar fashion. This effect did not appear to be due merely to the state of dryness of the sludge samples because in humus-deficient soil, a relatively inert material, the rate of poliovirus inactivation by heat was not significantly altered through dewatering. Instead, this effect appeared to have been caused by protective substances in the sludge, such as detergents, which are concentrated through dewatering. As reported previously (R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol. 34:681-688, 1977; R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol 36:889--897, 1978) raw sludge is not protective of reovirus, but, instead, the ionic detergents in sludge cause the rate of heat inactivation of this virus to be accelerated. Dewatering of sludge, however, was found to partially reverse this virucidal effect. Evidence is presented indicating that this reversal is caused by an unidentified protective substance in sludge also concentrated through dewatering. Finally, it was shown that the effects of raw sludge on heat inactivation of poliovirus and reovirus are greatly reduced by composting, a result that correlated with the degradation of detergents.


Membrane filter enumeration method for Clostridium perfringens.

A membrane filter procedure has been developed for the rapid quantitation of C. perfringens in the aquatic environment. Background growth is inhibited by the use of D-cycloserine, polymyxin B sulfate, and incubation at 45 degrees C. Differential characteristics include the fermentation of sucrose, production of acid phosphatase, and the absence of beta-D-glucosidase activity. The medium is prepared as follows (in grams per 100 ml of distilled water): tryptose, 3.0; yeast extract, 2.0; sucrose, 0.5; L-cysteine, 0.1; MgSO4. 7H2O, 0.01; bromocresol purple, 0.004; and agar, 1.5. The ingredients are dissolved, and the pH is adjusted to 7.6. After autoclaving at 121 degrees C for 15 min, the medium is allowed to cool at 50 degrees C, and the following are added per 100 ml: D-cycloserine, 40 mg; polymyxin B sulfate, 2.5 mg; indoxyl-beta-D-glucoside, 60 mg; 2.0 ml of a filter-sterilized 0.5% phenolpthalein diphosphate solution; and 0.2 ml of a filter-sterilized 4.5% FeCl3.6H2O solution. Enumeration of C. perfringens in a water sample is completed within 18 to 24 h. The verification of typical colonies was 93%. The average recovery from peptone-water spore suspensions of five strains was 79%, and that from filter-sterilized seawater suspensions was 90%. The precision of the method was approximately equal to that expected from random error alone. Confirmed recoveries of C. perfringens from water and sewage samples generally were greater than those by the Bonde pour tube method.


[Tumour regression in pituitary adenoma by bromocriptin (author's transl)].

A 35 year old patient with longstanding amenorrhea-galactorrhea due to a pituitary macroadenoma has been observed for a period of more than 2 years. During this time tumour expansion was radiologically evident. A full term pregnancy was responsible for most of the tumour growth. Following postpartum period Bromocriptin treatment led to considerable regression of the adenoma. Recalcification of sella structures and an involuted sella volume was radiologically evident. With reference to the experimental investigations of Lloyd (1975) and following the suggestions of L'Hermite (1977) and Vaidya (1977) this tumour regression is interpreted as being due to the antimitotic effect of Bromocriptin via inhibition of c-AMP and DNA.


Jaundice after renal allotransplantation.

Of 567 patients receiving renal transplantation at the University of Minnesota between October 1967 and October 1975, 22 developed clinical jaundice. Of these 22, nine died with their initial episode of hepatitis, six died within three months of causes associated with liver malfunction, four developed evidence of chronic hepatic failure and only three totally recovered from their illness. Five had clear evidence of Australia antigen positive hepatitis B, four of cytomegalovirus hepatitis, two of herpes hominis hepatitis, one of varicella zoster hepatitis and three of hepatic failure associated with systemic bacterial and/or fungal sepsis. Two of the 22 patients were thought likely to have cytomegalovirus hepatitis though definite proof was absent and in five patients a clear-cut etiology could not be made. In many of these patients the diagnosis was confounded by the previous presence of HB(s)Ag antigen and the frequent occurrence of a previous or concurrent infection with cytomegalovirus. The role of various drugs including azathioprine, sulfisoxazole, chlorpromazine, acetominophen, etc., could not be established but major roles for these agents in the face of the many viral and bacterial infections present in these patients is doubted. No clear-cut therapy could be established although it appears safe to discontinue azathioprine for longer or shorter periods of time with or without substitution of cyclophosphamide without serious deterioration of renal function. The problem of hepatic failure in transplant patients is still unsolved and will require a prospective study of etiologic agents and sub-clinical hepatic dysfunction in order to establish even the first principles of clinical-pathological correlation.


Action of p-(4-amidino-phenoxy)-benzaldehyde-p-amidino-phenylhydrazone dihydrochloride on Leishmania donovani infections in the golden hamster.

The chemotherapeutic effect of a new diamidine, HOE 668, the p-(4-amidino-phenoxy)-benzaldehyde-p-amidino-phenylhydrazone dihydrochloride, was compared with that of known anti-leishmanial drugs in golden hamsters infected with Leishmania donovani. The effect of HOE 668 against visceral leishmaniasis proved superior to that of pentamidine isethionate and the pentavalent antimonial drugs, sodium stibogluconate and N-methylglucamine antimoniate. However, HOE 668 can be used only experimentally because of its toxicity. Its very good anti-leishmanial action qualifies HOE 668 as a standard compound in screening tests.


The interaction between Entamoeba histolytica and Syphacia obvelata infection in mice.

Mice naturally infected with Syphacia obvelata were challenged intracaecally with Entamoeba histolytica trophozoites. They showed a higher level of amoebic infection than mice without worms. Mice with the heaviest Syphacia infection were the most susceptible to amoebic infection. It appears that a concomitant infection with helminthic parasites residing in the caecum alters the outcome of an amoebic infection.


Action of phenylbutazone, cyclophosphamide and prednacinolone on pleurisy due to Bordetella pertussis hypersensitivity in the rat.

Phenylbutazone, cyclophosphamide and prednacinolone acetonide, administered around the period of challenge reduced the exudate of pleurisy due to Bordetella pertussis hypersensitivity in the rat. The results on the leukocytes of the exudate are different depending on the class of product studied. Phenylbutazone only slightly reduced the number of mononuclears cells. Both the immunosuppressive and the corticoid induced a clear decrease in the total leukocyte number. But, while prednacinolone effects were nearly equally distributed among mononuclears and polynuclears, cyclophosphamide was mainly active on mononuclears.


Increase in urinary excretion of phosphates, hydroxyproline, and cAMP during acetate desoxycorticosterone + saline treatment in the rat.

We investigated the urinary excretion of phosphates, hydroxyproline, and cAMP in rats treated with desoxycorticosterone acetate (DOCA) associated with a hypersaline diet. We measured the three substances in groups with or without parathyroid or thyroid glands, or both. Urinary phosphate excretion increased greatly during the first weeks of treatment in normal rats and in rats autografted with parathyroids and lacking thyroid, and increased slightly in the groups without parathyroids. Hydroxyproline and cAMP had the same pattern; their excretion was greatest during week 2 of treatment in all groups, but these phenomena were more intense in animals with parathyroids. These fact support the conclusion that the parathyroid glands are stimulated during the start of mineralocorticoid-saline treatment.


Progesterone therapy in obese patients with sleep apnea.

Seven morbidly obese patients with sleep-related upper airway obstruction (UAO) were studied before and during progesterone treatment. All subjects had severe daytime somnolence, and five of the seven had congestive heart failure. All subjects were studied for one or two consecutive baseline nights in the sleep laboratory. Six of the seven subjects showed a mean of greater than 100 UAOs. Patients were treated for two to four weeks with medroxyprogesterone acetate, 20 mg three times daily. At the end of the treatment period, the sleep studies were repeated. There was no significant difference in the incidence, mean duration, or rate per minute of apneic episodes occurring before and with progesterone treatment. Severe hypoxemia during sleep persisted with treatment, as did the occurrence of cardiac arrhythmias. There was a significant improvement in the waking Pao2 with progesterone treatment.


Some observations on the opiate peptides and schizophrenia.

With the discovery of the opiate peptides, several major avenues of research became apparent. These peptides produced a great deal of focused attention on their anatomy, biochemistry, and physiology. In this article, we present an overview of some of the main research issues and recent findings in the field of opiate peptides. The possible relationship of the opiate peptide neuronal systems to schizophrenia is discussed in light of attempts to alter schizophrenic symptoms with opiate antagonists, beta-endorphin, and dialysis. It is hypothesized that if the opiate peptides are involved in schizophrenia, then their involvement with dopamine systems and/or with stress responses may be critical.


Successful separation of depressed, normal, and insomniac subjects by EEG sleep data.

Data from all-night EEG sleep studies were used to distinguish normal subjects, primary depressed patients, and primary insomniac patients. In part 1, we compared 41 normal subjects, 56 depressed patients, and 18 insomniacs. In a univariate comparison with normal subjects, depressed patients showed less total sleep, longer sleep latency, more early morning awake time, more intermittent awake time, less delta sleep, less sleep efficiency, and shorter rapid eye movement (REM) latencies; compared with insomniacs, depressed patients showed greater early morning awake time, shorter REM latency, greater REM index, and greater REM density. Using multivariate discriminant analysis, 82% of the sample were correctly classified by diagnosis: 100% of the normal subjects, 72% of the depressed patients, and 77% of the insomniacs. Eight variables contributed to the multivariate separation of depressed individuals from insomniacs and normals: total sleep time, total recording period, sleep efficiency, sleep latency, early morning awake time, awake time, REM time and REM%. When the discriminant functions were applied to a second group of 18 primary depressed patients, 82% were correctly classified as depressed. These results suggest that primary depressed patients and primary insomniac patients may show relatively characteristic patterns of sleep abnormality.


[Monoamine metabolites and cyclic nucleotides in the cerebrospinal fluid of patients with bismuth or mercury poisoning].

The central metabolism of dopamine, serotonin, cyclic AMP and cyclic GMP was studied by use of the probenecid test in three patients with bismuth encephalopathy and in one patient with mercury encephalopathy. The accumulation of HVA and of cGMP in the cerebrospinal fluid was depressed during the acute phase of bismuth encephalopathy with severe hyposomnia, while it was increased in a patient with regression of clinical symptoms and normal in a patient with more advanced recovery. The patient with chronic mercury poisoning showing a severe cerebellar ataxia and rigidity had an almost complete suppression of HVA accumulation and an increase of cGMP accumulation. No pronounced differences of 5-HIAA and cAMP behavior were found. It is concluded that the central metabolism of dopamine and of cGMP is severely affected in bismuth and mercury encephalopathies.


The effect of dextransulfate 500 on the pathogenesis of herpes simplex virus infections in weanling mice.

Intraperitoneal (i.p.) injection of Dextran Sulfate (D.S.) 500 during a limited period of time influences the course of herpes simplex-virus-infections. D.S.500 was found to reduce the resistance of mice for some herpes simplex-virus strains (Len, L3--2s, Haase) if given between 16 hours and 2 hours after i.p. infection. The decrease of resistance could be correlated with an increase of the virus content of liver, spleen, brain and spinal cord. Injection of herpes simplex-virus-specific immune serum counteracted the effect of D.S.500 on the course of infections. Conversely, D.S.500 increased the resistance of mice to another group of herpes simplex-viruses (strains D-316, Thea, DD), if given 3 to 8 hours before infection. These effects are ascribed to a special interaction of D.S.500 with macrophages and probably other virus-susceptible cells of the peritoneal cavity and elsewhere with a resulting counteraction to the virus infection.


Stimulation of host DNA synthesis and induction of early antigens by ultraviolet light irradiated human cytomegalovirus.

The ultraviolet (UV-)sensitivity of the human cytomegalovirus (HCMV) genes coding for very early complement fixing and early antigens in human embryonic fibroblasts (HEF) and mouse embryonic fibroblasts (MEF) and the relation of these genes to the ability of the virus to stimulate host cell DNA synthesis were investigated. After 14 minutes of UV-irradiation of the virus inoculum only the very early complement fixing nuclear antigen (CMNA) developed in the HEF cells and only the early cytoplasmic antigen(s) was present in the MEF. In both HEF and MEF, host cell DNA synthesis was stimulated. We conclude that the ability of HCMV to stimulate host DNA synthesis is an early function of the viral genome and shows a high resistance to UV-irradiation. There is no direct correlation, however, between the ability of the virus to stimulate host cell DNA synthesis and the genes which code for the CMNA or for early cytoplasmic antigens.


Reversible inhibition of the induction of DNA polymerase of herpes simplex virus type 2 in HeLa cells.

The induction of DNA polymerase of herpes simplex virus type 2 in HeLa cells was inhibited by 2-mercapto-1-(beta-4-pyridethyl)benzimidazole added at an early time after infection. However, the inhibitory effect was easily released by removal of the inhibitor. Since the drug disorganizes nucleolar structure and prevents its function in HeLa cells in a reversible manner, it is possible that nucleoli are involved in the induction of DNA polymerase of this virus.


[Patterns of cytomegaly generalization and its pathomorphological diagnosis].

On the basis of morphological examinations of 576 cases of cytomegalia, several summarizing statements concerning the regularities of its generalization are presented. According to the nature of tissue reactions in the salivary glands which correlatively depend on the status of immunogenesis organs and the form of the disease, 4 morphological types of cytomegalic sialoadenitis are distinguished: nodular, immunodeficient, persisting, and hemopoietic. It is recommended that the discovered regularities be taken into consideration in autopsy practice which will permit to minimize the cases of unrecognized generalization of the infection.


[Histochemical characteristics of the mucus in pseudomyxoma peritonei].

Histochemical investigations of gelatinous masses of 3 patients with pseudomyxoma peritonei were carried out. The tissues of the mucinous adenocarcinoma of the ovary and normal appendix were used for comparison. The results showed that the mucus in pseudomyxoma peritonei as well as that of the epithelial cells of the appendix belonged to the group of acid nonsulfurated mucoid substances of epithelial origin rich in syalic acid. The mucus in the mucinous adenocarcinoma of the ovary had the same histochemical properties, except for its resistance to neuraminidase. Some suggestions on the pathogenetic mechanisms of the development of the disease and possibilities of therapeutic applications of neuraminidase are presented.


[Mucoepidermoid tumor of the epipharynx].

Case report about a mukoepidermoid-tumor, which was investigated by means of histology and electron microscopy. The localisation of this tumor in the epipharynx is a rarity and unknown in the literature. The variable differentiated tumor consists of epidermoid cells with different graduation of maturity, of epithelial cells from the gland ducts and primitive acini as well as in the majority of undifferentiated cell types. The tumor seems to be of a "low grade malignity".


Coxsackie myocarditis complicating treatment of Hodgkin's lymphoma.

A fatal case of Coxsackie B4 myocarditis complicating treatment of Hodgkin's lymphoma in an adult is reported. The virus was isolated from the myocardium obtained at autopsy and light microscopy confirmed a myocarditis with involvement of the conduction system. Electron-microscopy showed dense mitochondrial inclusions. Coxsackie myocarditis rarely results in death of affected adults. It is postulated in this case that the treatment of the Hodgkin's lymphoma resulted in B lymphocyte depletion, allowing a more virulent infection to occur with resultant fatal myocarditis.


Pitfalls in carpal tunnel decompression.

In the period 1972 to 1977, 35 patients with persistent pain and other disabilities after carpal tunnel decompression were referred for treatment. The pitfalls included: (i) wrong diagnosis--5 patients; (ii) inadequate decompression--9 patients; (iii) recurrent compression--4 patients; (iv) iatrogenic nerve injury--7 patients; and (v) postoperative complications--10 patients. The patients in Groups 1 and 5 were treated by non-operative measures. Most of the patients in Groups 2, 3, and 4 were treated by reoperation. This paper outlines the causes and prevention of pitfalls in carpal tunnel decompression.


Esterases of Mus musculus: substrate and inhibition characteristics, new isozymes, and homologies with man.

A wide range of fluorogenic and naphthol esters has been tested as substrates for mouse esterases. New esterases have been identified in liver and kidney extracts with palmityl, oleyl, and elaidyl esters. From substrate, inhibition, and molecular weight studies, three homologies between human and mouse esterases are suggested. A new allele at Es-6 is also described.


Intrasubunit nucleotide binding in ribonucleic acid polymerase.

1. Periodate oxidation of the ribose ring was used to synthesize derivatives of nucleoside triphosphates. 2. These oxidized nucleoside triphosphates. 2. These oxidized nucleoside triphosphates are competitive inhibitors of RNA polymerase. 3. On incubation, together with NaBH4, these oxidized labelled nucleotides are covalently bound to Escherichia coli RNA polymerase. 4. Nucleoside triphosphate substrates decrease the extent of labelling. 5. A lysine residue in an alpha-subunit is labelled. 6. The significance of these results in relation to the location of the nucleotide-binding site is discussed.


alpha-L-iduronate ring conformations in heparin and heparin derivatives. 13-C Nuclear-magnetic-resonance analysis and titration data for variously desulphated and periodate-oxidized heparins.

A heparin derivative that had been O/N-desulphated and re-N-acetylated was investigated by 13C n.m.r. spectroscopy and potentiometric titration. Three forms of uronic acid were observed, tentatively identified as beta-D-glucuronate, and two different forms of alpha-L-iduronate. A comparison of the n.m.r. spectra of heparin, an oligosaccharide (beta-D-glucuronate-2-acetamido-2-deoxy-alpha-D-glucose)n, and heparin that had been subjected to selective oxidation of beta-D-glucuronate, enabled the position of the anomeric carbon of the latter residue to be assigned [delta 102.9 (p.p.m.)]. Periodate oxidation of O/N-desulphated heparin destroyed in addition, approx. 40% of the alpha-L-iduronate content. The remainder of the alpha-L-iduronate residues displayed only one anomeric resonance, at delta 99.7 (p.p.m.). In another preparation, after sequential desulphation of heparin (N-desulphation, re-N-acetylation and O-desulphation) the anomeric resonance of the alpha-L-iduronate residue shifted downfield [from delta99.7 (p.p.m.) to delta 102.3]indicating a change in ring conformation. These data support the interpretation that the unsulphated alpha-L-iduronate residues may adopt two conformations. It was shown that the proportions of alpha-L-iduronate conformers are determined by the sequence of desulphation operations. Also minor components of heparin were assigned.


Action of liproprotein lipase on apoprotein-depleted chylomicrons.

1. Rat lymph chylomicrons were exposed to soluble and to immobilized trypsin. This treatment caused no detectable changes in the chylomicron structure or lipid composition, but did result in virtually total depletion of all their tetramethylurea-soluble apoproteins. 2. The capacity of these apoprotein-depleted chylomicrons to act as substrate for lipoprotein lipase in vitro and in situ (i.e. isolated perfused rat heart) was decreased by about 90 and 75% respectively, compared with intact chylomicrons. 3. On incubation with rat plasma high-density lipoproteins, trypsin-treated chylomicrons readily acquired a full apoprotein complement. This resulted in the complete restoration of their capacity to act as substrate for lipoprotein lipase both in vitro and in situ. 4. It is suggested that with the use of try,sin-treated chylomicrons it is now possible for the first time to investigate the physiological role that individual apoproteins play in the catabolism of triacylglycerol-rich lipoproteins by lipoprotein lipase.


Interaction of Sendai virus with human erythrocytes. IV - Hemolytic factors.

A study on the composition of structural proteins of Sendai virus deprived of hemolytic activity by pyridine and trypsin in controlled condition was carried out. Pyridine leaves unaffected the electrophoresis pattern of virus proteins; in virus treated with trypsin a drastic reduction of F protein was observed. Researches reported indicate that two factors are required for hemolytic activity of Sendai virus the F protein and the phospholipids of the outer layer of envelope.


Specific cellular unreactivity during cytomegalovirus infection in man.

The specific cellular immune response against Cytomegalovirus (CMV) was studied by the Leucocyte Inhibitory Factor (LIF) production test in three groups of subjects: 7 healthy individuals with no CMV antibodies, 4 with antibodies to CMV but without evidence of active infection, and 2 children with clinical CMV infection. Our results show that LIF production in presence of CMV is high in normal seropositive subjects, while it is strongly deficient in patients with active infection, despite a good humoral response to the virus. In one child with congenital infection, a low number of E rosette forming cells was also revealed. We suggest that CMV infection can induce, at least during infancy, a state of specific cellular unresponsiveness to the virus. When the infection is congenital, a generalized depression of the cellular immune system may develop.


Reactivities of systemic lupus erythematosus sera with cellular and virus antigen preparations.

Sera of patients with systemic lupus erythematosus (SLE) showed significantly higher complement fixation reactivities with 8 of 10 virus antigen preparations compared to sera of normal subjects, including virus workers, confirming earlier observations. However, one-third of SLE sera were also reactive with cellular antigens, prepared in a manner identical to viral antigens, but from cells not infected with virus. By contrast, only one of 49 normal sera showed reactivity with one of nine cellular antigens, at a minimal titer. Apparently heightened reactivities of SLE sera with virus antigens could be explained in many instances on the basis of reactions with cellular antigens. Serum reactivities with nonviral tissue antigens in SLE must be considered in interpretation of immunologic studies related to viruses and SLE.


Prevalence of antibody to hepatitis A virus in healthy individuals of Milan.

We have studied by radioimmunoassay the prevalence of antibody to hepatitis A virus in 436 randomly selected individuals of Milan according to age and sex. Taken together, 70% of subjects provided serological evidence of exposure to HAV. Both sexes resulted similarly immunized (69.7% females and 70.7% males). The anti-HAV prevalence was found to be directly related to age, being on the increase from childhood to adulthood and reaching its peak (9l.5%) in people over fifty. An analysis of the distribution of anti-HAV according to presence or absence of anti-HBs in induviduals under 30 years, revealed that anti-HAV was detectable more frequently in individuals anti-HBs positive than in those without such evidence in sera (71.4% versus 39.7%; P less than 0.01). Finally, 96 sera from children and adolescents of low socioeconomic standard of living were also investigated. In these subjects anti-HAV prevalence was found to be higher than in their counterparts randomly slected from general population.


Search for type-C oncornavirus-related genetic information in tissues from patients with systemic lupus erythematosus.

Single-stranded 3H-DNA probes complementary to the RNA of Rauscher murine leukemia virus and of simian sarcoma virus were prepared using techniques that permitted complete transcription of the viral genome of each virus. These probes were used in DNA-DNA hybridization studies with the cellular DNA from uncultured specimens of spleens and placentas of patients with systemic lupus erythematosus (SLE). No proviral DNA sequences related to these viruses were detected in these tissues. The results presented here do not support previously reported antigenic data implicating type-C oncornavirus infection of these organs in SLE.


Seroepidemiologic study of human coronavirus OC43 infections in Italy.

Human Coronavirus OC43 from suckling mouse brain (SMB) suspension has been adapted to grow in human embryonic lung fibroblast cell cultures. Complement-fixing and hemagglutinating antigens were prepared from infected cell cultures and used in parallel with the same antigens from SMB for a study of the prevalence of OC43 antibody in Northern Italy as well as for the serodiagnosis of recent OC43 infections and reinfections. Results show (i) a higher sensitivity of the hemagglutination-inhibition as compared to the complement fixation test both for antibody detection in seroepidemiological studies and serodiagnosis of acute infections, (ii) a high prevalence of OC43 antibody in Northern Italy, so that only exceptionally are people older than 12 years seronegative, (iii) a high rate of OC43 reinfections during the winter-spring season 1976/77 in a population sample with preexisting antibody (iv) a good quality of reagents prepared from cell cultures, and hence free of contaminating mouse viruses, such as mouse hepatitis virus.


Pharmacokinetics and pharmacological effects of neostigmine in man.

1 The pharmacokinetics of neostigmine was studied in six patients during the reversal of neuromuscular block induced by tubocurarine chloride. The effect of the drug on neuromuscular function was simultaneously assessed by electromyography. 2 Neostigmine was rapidly eliminated from plasma after intravenous administration. The decline in the plasma concentration of the drug was invariably resolved into two exponential components. The fast disposition (distribution) half-life of the drug was invariably less than 1 min; the slow disposition (elimination) half-life ranged from 15.4--31.7 min. 3 Neostigmine usually increased the amplitude of the compound muscle action potential and diminished electromyographic decrement within 2 min of intravenous injection. The pharmacological effect of neostigmine was usually maximal between 7 and 15 min. There was an inverse relationship between the plasma concentration of the drug and the facilitation of neuromuscular transmission. 4 Red cell acetylcholinesterase activity was almost completely inhibited within 2--3 min of intravenous injection of neostigmine. Enzyme activity recovered to approximately 28% of control values by 30 min and to 55% by 60 min.


Parapoxvirus infections of reindeer and musk ox associated with unusual human infections.

This report concerns five cases of human infection resulting in lesions morphologically similar to human orf. Parapoxvirus infection in reindeer was the probable source of infection in four cases. The fifth case involved musk ox. Two of the cases involving reindeer are particularly interesting as transmission of infection occurred indirectly. Viral particles were not seen by electron microscopy of human tissues, due probably to the late stage of the illness at the time of examination. However, electron microscopy of negatively stained suspensions prepared from biopsy material collected from reindeer calves at the outbreak of the illness and of post-mortem material from the musk ox revealed characteristic parapoxvirus particles. In one patient the infection was accompanied by fever, lymphadenopathy and nausea, which cleared following a few days of treatment with doxycycline chloride. 4 cases were curetted after 4, 6, 12 and 20 weeks and healed withour scarring whereas the fifth patient was by request left alone. This resulted in localized dark pigmentation following completion of the healing process after 26 weeks.


Plasma levels of oestrone, oestradiol and gonadotrophins in postmenopausal women after oral and vaginal administration of conjugated equine oestrogens (Premarin).

Peripheral plasma from five postmenopausal women was analyzed for oestrone and oestradiol during 48 hours after administration of 1.25 mg Premarin (conjugated equine oestrogen) orally and vaginally. Oestrone and oestradiol were separated on columns of Sephadex LH 20 before radioimmunoassay. Administration of 1.25 mg of Premarin produced a marked increase in plasma oestrone levels and 24 hours after treatment the oestrone levels were still above follicular phase levels. The plasma pattern of oestradiol was similar but the elevation was less pronounced. The biological activity of Premarin as indicated by depressed gonadotrophin levels was similar with the intravaginal and oral routes of administration.


Failure to suppress adrenal function in congenital adrenal hyperplasia (21-hydroxylase deficiency). Three case reports.

Three patients, aged 14, 16, and 32 years respectively, with congenital adrenal hyperplasia (21-hydroxylase deficiency) are described. Excessive adrenal activity and ACTH secretion could not be suppressed with doses of corticosteroids sufficient to cause iatrogenic Cushing's syndrome, even though part of the steroid dosage was administered in the late evening. The resistance to feed-back suppression was of the same order as that seen in Cushing's syndrome. Adrenalectomy was performed in the 16-year-old girl, and was followed by a menarche. Adrenalectomy was considered inadvisable in the other two patients.


Adenovirus type 21 keratoconjunctivitis.

A case of keratoconjunctivitis caused by adenovirus type 21 in London has been described. A 59-year-old woman attented hospital in August 1974 complaining of a 3-week history of redness, grittiness, watery discharge, and photophobia in her left eye and a slight upper respiratory infection. Clinical examination showed a moderate follicular conjuctivitis mainly in the lower and upper fornices, which lasted for 6 weeks. In the cornea a moderate amount of epithelial and subepithelial punctate keratitis was observed. The subepithelial opacities were coarse, discrete, and round and lasted for 4 months. The course of follicular conjunctivitis and the subepithelial punctate keratitis in this patient was similar to epidemic keratoconjunctivitis caused by adenovirus 8. A conjunctival swabbing collected from this patient was positive for adenovirus serotype 21.


Inhibition of membrane fusion by suppression of lateral movement of membrane proteins.

Chicken erythrocytes were fused either by Sendai virus or by the combination of Ca2+ and ionophore A23187. Intramembrane particles and external anionic sites of cells undergoing fusion were found to acquire the ability to undergo a process of cold-induced clustering (thermotropic separation). Cationized ferritin (200 microgram/ml 5% (v/v) cell suspension) inhibited both the fusion process and the thermotropic separation of intramembrane particles and external anionic sites. The correlation between the mobility of membrane proteins and the fusion process is discussed. It is suggest that an increase in the lateral mobility of membrane proteins is a prerequisite for initiation of membrane fusion.


Streptomycin-induced conformational changes in the 70-S bacterial ribosome.

Conformational alterations induced by streptomycin in the bacterial ribosome have been investigated using as probes, ethidium bromide, N-[14C]ethylmaleimide and a spin label nitroxide analog of N-ethylmaleimide. 1. The binding of the antibiotic to the ribosome does not affect the reactivity of sulfhydryl groups towards N-ethylmaleimide. 2. The motional freedom of spin labels bound to ribosomal proteins S1 and S18 is increased but it is hardly affected at other labeled sites. This observation suggests that the binding of streptomycin causes a local loosening of the ribosomal structure. 3. Ribosomes are found to bind less ethidium bromide in the presence of streptomycin, which suggests that the binding of streptomycin decreases the degree of organization of ribosomal RNA.


Non-histone chromatin proteins of B lymphocytes stimulated by lipopolysaccharide. II. Phosphorylation.

Stimulation of mouse lymphocytes with the B lymphocyte specific mitogen lipopolysaccharide results in an increased rate of phosphorylation of non-histone chromatin proteins. An initial small increase in phosphorylation occurs during the first 2 h and a much larger increase after 24 h of culture with mitogen. The phosphorylated nuclear and cytoplasmic proteins were analysed by polyacrylamide gel electrophoresis and the stimulation index of each prominent peak measured. It was inferred that selective stimulation of the phosphorylation of individual proteins had occurred from: (1) the range of stimulation indices for different proteins, and (2) the appearance, after 8 h stimulation of an apparently newly phosphorylated non-histone chromatin protein of molecular weight 115 000. The pool size of ATP was monitored and showed only small changes during the first 24 h of exposure to lipopolysaccharide. Phosphatase activity was found to be associated with lymphocyte chromatin and nucleoplasm and may help to regulate the level of phosphorylation of non-histone chromatin proteins in vivo. To preserve phosphorylated proteins during their isolation phosphatase activity was inhibited by Na2MoO4. The selective changes in phosphorylation of nuclear proteins precede, and continue during, the stimulation of immunoglobulin and DNA synthesis. Our results are thus consistent with the hypothesis that phosphorylation of non-histone chromatin proteins plays a role in the regulation of gene expression in B lymphocytes.


Induction of mitochondrial phosphoenolpyruvate carboxykinase in cultured human fibroblasts.

Mitochondria of cultured normal human fibroblast cells were found to contain the enzyme phosphoenolpyruvate carboxykinase. The activity of this enzyme in these cells is increased 2- to 3-fold by addition of 5 . 10(-4) M dibutyryl cyclic AMP, or 1.5- to 2-fold by the addition of dexamethasone (2 . 10(-7) M) or hydrocortisone (1.38 . 10(-6) M). These increases in enzyme activity were inhibited cycloheximide and actinomycin D, suggesting they are dependent upon de novo protein synthesis. Cultured human fibroblasts may thus provide a useful system for studying the regulation of mitochondrial phosphoenolpyruvate carboxykinase.


Characterization of phosphatidate phosphohydrolase activity associated with isolated lamellar bodies.

It has been demonstrated that lamellar bodies isolated from porcine lung have L-alpha-phosphatidate phosphohydrolase (EC 3.1.3.4) activity which cannot be accounted for by microsomal or mitochondrial contamination. Phosphatidate phosphohydrolase activity associated with the lamellar bodies is relatively insensitive to Mg2+ and more heat labile than the activity associated with whole lung microsomes. The enzyme was found to be the most active phosphohydrolase present in isolated lamellar bodies and is inhibited by 5 mM Be2+. Lamellar bodies isolated from human and rat lung tissue were also found to have this activity, and the functional role of the enzyme in lamellar bodies is proposed in relation to glycerophospholipid metabolism.


Evidence for the existence of a single enzyme catalyzing the phosphorylation of choline and ethanolamine in primate lung.

Choline kinase (ATP:choline phosphotransferase, EC 2.7.1.32) has been isolated and purified 1000-fold from adult African Green monkey lung with a yield of 10%. The purified enzyme also phosphorylated ethanolamine (ratio of ethanolamine kinase to choline kinase = 0.30). This ratio remained constant throughout the purification procedure. The Km for choline (3.0 - 10(-5) M) was lower than that of ethanolamine (1.2 - 10(-3) M.) Choline was also found to inhibit ethanolamine kinase activity by 50% at a concentration of 0.005 mM, while ethanolamine inhibited choline only at very high concentrations (100--150 mM). When the enzyme was subjected to inactivation by heat, hemicholinium-3, trypsin digestion, and p-hydroxymercuribenzoate, both ethanolamine kinase and choline kinase activities were destroyed at the same rate. Freezing and thawing in the absence of glycerol also destroyed both activities at the same rate. Based on these findings, we conclude that in adult African Green monkey lung tissue, there is only one enzyme for the phosphorylation of ethanolamine and choline, and that choline phosphorylation predominates.


Heterogeneity in the low density lipoproteins of cholesterol fed African green monkeys (Cercopithecus aethiops).

The low density lipoproteins of cholesterol-fed African green monkeys were isolated by either preparative ultracentrifugation or agarose column chromatography, then radioiodinated and fractionated by agarose column chromatography. A marked variation was noted in the specific radioactivity of the column fractions regardless of the method used to obtain low density lipoproteins. The radioactivity in each fraction migrated with the beta-globluins, but when the fractions were mixed with carrier plasma, alpha-migrating radioactivity was also detected which was associated with the apo-A-I and apo-C of the high density lipoproteins. The uptake of these apoproteins by the high density lipoproteins reduced the variation in specific radioactivity among the low density lipoprotein fractions. By successive injections of the various 125I-labeled low density lipoprotein fractions into the same recipient, it was also possible to compare the kinetic behavior of the fractions. Large and small low density lipoprotein particles were metabolized differently, indicating that the low density lipoproteins from cholesterol-fed African green monkeys are heterogeneous.


Comparison of the phospholipase activity of bovine milk lipoprotein lipase against rat plasma very low density and high density lipoprotein.

The hydrolytic activity of a lipoprotein lipase from bovine milk against triacylglycerol and phosphatidylcholine of rat plasma very low density lipoprotein was determined and compared to that against phosphatidylcholine of high density lipoprotein. 85--90% of the triacylglycerol in very low density lipoprotein were hydrolyzed to fatty acids and 25--35% of the phosphatidylcholine to lysophosphatidylcholine. High density lipoprotein phosphatidylcholine was only minimally susceptible to the enzyme. Even with high amounts of enzyme and prolonged incubation periods, lysophosphatidylcholine generation did not exceed 2--4% of the original amounts of labeled phosphatidylcholine in the high density lipoprotein. We conclude that phospholipids in high density lipoprotein are not substrates for the phospholipase activity of this lipoprotein lipase. These observations suggest that factors other than the presence of apolipoprotein C-II and of glycerophosphatides are of importance for the activity of lipoprotein lipases.


Beta-oxidation in peroxisomes of brown adipose tissue.

Peroxisomes and mitochondria from brown adipose tissue of the rat were separated by differential pelleting and isopycnic gradient centrifugation. Both fractions oxidized palmitoyl-CoA with comparable specific activities. Unlike the mitochondrial beta-oxidation the peroxisomal activity was not influenced by carbon monoxide. Peroxisomal beta-oxidation together with carnitine acetyl-transferase, which is also located in peroxisomes, might be involved in chemical thermogenesis by delivering acetyl groups to the mitochondria.


Electrogenic events in the ubiquinone-cytochrome b/c2 oxidoreductase of Rhodopseudomonas sphaeroides.

The reductant of ferricytochrome c2 in Rhodopseudomonas sphaeroides is a component, Z, which has an equilibrium oxidation-reduction reaction involving two electrons and two protons with a midpoint potential of 155 mV at pH 7. Under energy coupled conditions, the reduction of ferricytochrome c2 by ZH2 is obligatorily coupled to an apparently electrogenic reaction which is monitored by a red shift of the endogeneous carotenoids. Both ferricytochrome c2 reduction and the associated carotenoid bandshift are similarly affected by the concentrations of ZH2 and ferricytochrome c2, pH, temperature the inhibitors diphenylamine and antimycin, and the presence of ubiquinone. The second-order rate constant for ferricytochrome c2 reduction at pH 7.0 and at 24 degrees C was 2 - 10(9) M-1 - s-1, but this varied with pH, being 5.1 - 10(8) M-1 = s-1 at pH 5.2 and 4.3 - 10(9) M-1 - s-1 at pH 9.3. At pH 7 the reaction had an activation energy of 10.3 kcal/mol.


Isolation of photosynthetic catalysts from cyanobacteria.

Methods are described for the isolation of ferredoxins I and II, cytochrome c-553, cytochrome f, cytochrome c-550 and plastocyanin from large quantities of various cyanobacteria. The amino acid composition of cytochrome c-550 is reported. There is a variation in the relative amounts of these proteins in different batches of cells which may relate to the nutritional status of the organisms.


Studies of the orientation of the mitochondrial redox carriers. III. Orientation of the gx and gy axes of the hemes of cytochrome oxidase with respect to the plane of the membrane in oriented membrane multilayers.

The EPR absorption properties of the hemes of cytochrome oxidase and their liganded derivatives were examined in oriented multilayers from isolated oxidase, mitochondrial membranes and membrane fragments of a bacterium, Paracoccus denitrificans. The hemes of the oxidase in all the systems investigated were oriented normal to the plane of the multilayers. The directions of the g signals corresponding to the gx and gy axes of the g tensor were found to be different in low-spin ferric heme in fully oxidized oxidase and in half-reduced liganded oxidase. It is suggested that this different orientation of gx and gy in fully oxidized oxidase and half-reduced liganded oxidase arises because the respective EPR signals belong to two different hemes, those of cytochrome a and a3.


On the active transport of organic acids (fluorescein) in the choroid plexus of the rabbit.

The kinetics of active transport of an organic acid (fluorescein) through the membranes of the choroid plexus from the lateral ventricules of the brain of rabbit was studied both morphologically and functionally. It was shown that fluorescein is actively translocated through the apical and basal membrane of the epithelium and is accumulated in blood capillaries at a concentration exceeding one order of magnitude that in the incubation medium. The kinetic curves displaying saturation and the demonstration of inhibition by other acids shows that a specific carrier is involved in the transfer across the membrane. The active transport of fluorescein at 20 degrees C was found to be sodium independent. Total exclusion of sodium from the incubation medium does not change the Michaelis constant (Km) and maximal velocity (V). The active transport depends on the operation of (Na+ + K+)-ATPase as energy source but obviously no specific complexes with the participation of sodium are involved.


Muscle surface membranes: preparative methods affect apparent chemical properties and neurotoxin binding.

Considerable disagreement exists between results reported by various authors for lipid composition and enzyme activity in purified muscle membrane fractions presumed to be sarcolemma, although an explanation for these discrepancies has not been presented. We have prepared muscle light surface membrane fractions of comparable density (1.050--1.120) by a low-salt sucrose method and by an LiBr-KCl extraction procedure and compared them for density profile, total lipid and cholesterol content, protein composition and ATPase activity. In addition, sodium channels characteristic of excitable membranes have been quantitated in each preparation using [3H]saxitoxin binding assays, and the density of acetylcholine receptors determined in fractions from control and denervated muscle using alpha-[125I]bungarotoxin. Although both fractions contain predominantly surface membrane, the LiBr fraction consistently shows the higher specific activity of p-nitrophenylphosphatase, higher free cholesterol content, and higher density of sodium channels and acetylcholine receptors. The density distribution of sodium channels appears uniform throughout both fractions. Quantitative differences were seen between sodium dodecyl sulfate polyacrylamide gel electrophoresis patterns of membrane proteins from the two preparations although most bands are represented in both. A majority of the low-salt sucrose light membrane proteins were accessible in varying degrees to labelling with diazotized diiodosulfanylic acid in intact muscle. These results suggest that light surface membrane fractions may be mixtures of sarcolemma and T-tubular membranes. Using our preparative methods, the LiBr fraction may contain predominantly sarcolemma while low-salt sucrose light membranes may be enriched in T-tubular elements.


Metabolism of lipoprotein acylglycerols by liver parenchymal cells.

We investigated the metabolism by hepatocyte suspensions of the acylglycerols in lipoprotein remnants as well as those associated with albumin and low or high density lipoproteins. Remnants, albumin and plasma lipoproteins, rich in monoacylglycerol were prepared by short-term incubations of radio-labeled chylomicra or very low density lipoproteins with extrahepatic lipoprotein lipase in the presence of albumin and low and high density lipoproteins. We demonstrated that liver parenchymal cells contain an active monoacylglycerol acyltransferase that is located on the extracellular surface of the cell plasma membrane. Further, the enzyme is capable of degrading the monoacylglycerol in all the above forms. Triacylglycerol in intact chylomicra and very low density lipoproteins were not metabolized by the cells to any appreciable degree. The degradation of the remnant triacylglycerol appeared to depend solely on the activity of the lipoprotein lipase bound to the lipoprotein remnants. Little uptake of intact lipoprotein acylglycerols by the hepatocytes was observed; instead, hydrolysis of the substrates in the medium always preceded the uptake of the products. The products were then utilized for the synthesis of triacylglycerol and phospholipid within the cells.


Effects of betamethasone on phospholipid content, composition and biosynthesis in the fetal rabbit lung.

Administration of betamethasone (0.2 mg/kg, intramuscularly) to pregnant rabbits had the following effects on the fetal lung at 26--27 days gestation. It increased the amount of phosphatidylcholine in lung lavage by 70% and almost doubled the phosphatidylcholine/sphingomyelin ratio, it increased the rate of incorporation of choline into phosphatidylcholine in fetal lung slices by up to 90%, it increased the activities of pulmonary cholinephosphate cytidylyltransferase and phosphatidate phosphatase by 50% and it reduced the amount of lung glycogen to 60% of the amount in the controls. Betamethasone had no effect on the activities of pulmonary cholinephosphotransferase or lysolecithin: lysolecithin acyltransferase but it slightly decreased the activity of choline kinase. Betamethasone administration to the doe did not increase the amount of surfactant phospholipid in fetal lung lavage to as great an extent as did direct administration of cortisol to the fetuses. Neither did betamethasone stimulate the activity of pulmonary cholinephosphotransferase. These data suggest that agents other than glucocorticoids mediate the stress-induced acceleration of fetal lung maturation and surfactant production.


Analysis of electrophoretic behavior of cytochrome c oxidase subunits. Retardation coefficient versus molecular weight in dodecyl sulfate urea gels.

1. Standard proteins were examined by electrophoresis in highly cross-linked polyacrylamide gels containing sodium dodecyl sulfate and urea. Their behavior was analyzed at a single gel concentration (by molecular weight vs. relative mobility) and at several gel concentrations (molecular weight vs. retardation coefficient). The validity of the latter method of analysis was established for this gel system. 2. Cytochrome c oxidase was subjected to analysis by this method. Compared to standards, subunits I and III showed increased free electrophoretic mobility, while that of subunit V was slightly decreased. The molecular weight values derived were: I, 44 600; II, 22 700; III, 23 500; IV, 16 900; V, 9400; VI, 7600; VII, 4300. The standard errors were all less than +/- 7%. 3. Isolated V and VI were analyzed by two dimensional dodecyl sulfate electrophoresis, in which the second dimension also contained urea. In contrast to their behavior when the holo-enzyme was examined, these isolated subunits V and VI no longer exchange migrating positions relative to each other during the two dimensional analysis. The molecular weight values of the isolated subunits agree with those of the holo-enzyme.


Properties and purification of a glucose-regulated protein from chick embryo fibroblasts.

A glucose-regulated protein of molecular weight 78,000 (GRP-78) had been purified from a membrane fraction isolated from viral transformed chick embryo fibroblasts. Purification was achieved by extraction of the membrane fraction with Triton X-100, and chromatography on diethylaminoethyl-cellulose and hydroxyapatite. The purified protein exhibited one single spot on two-dimensional polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and has a pI of about 5.3. A monospecific antiserum to GRP-78 was generated in a goat. Immunofluorescence studies using affinity purified antibodies to GRP-78 revealed that this protein was not exposed on the cell surface but was localized in a granular vesicular network inside the cell that resembles the distribution of endoplasmic reticulum. The availability of purified GRP-78 and a specific antiserum to it should prove useful in elucidating the role of this protein in glucose metabolism and its relationship to malignant transformation.


Biosynthesis of enterochelin in Escherichia coli K-12: separation of the polypeptides coded for by the entD, E, F and G genes.

Four enzymic components, coded for by the entD, entE, entF and entG genes, involved in the biosynthesis of enterochelin from 2,3-dihydroxybenzoate have been separated from cell extracts of mutant strains of Escherichia coli K-12. The starting material for fractionation of the E, F and G components was a cell extract of an entD mutant strain, which yielded the E, F and G enzymic components uncontaminated by a functional D component. The D component was isolated from cell extracts of an entE mutant strain. The conversion of 2,3-dihydroxybenzoate and L-serine into enterochelin is dependent on the presence of all four enzymic components. The E and F components were shown to catalyze ATP-pyrophosphate exchange reactions dependent on 2,3-dihydroxybenzoate and L-serine, respectively, whereas fractionated extracts of the entE and entF mutant strains lacked these reactions. These data provide firm evidence that the E and F components are involved in the initial activation of the substrates. The D and G components are necessary for subsequent and, as yet, undefinedd reactions.


Regulatory role of cyclic adenosine 3',5'-monophosphate on the platelet cyclooxygenase and platelet function.

Thromboxane A2 plays an important role in arachidonic acid- and prostaglandin H2-induced platelet aggregation. Agents that stimulate platelet adenylate cyclase (prostaglandin I2, prostaglandin I1 and prostaglandin E1) and dibutyryl cyclic AMP inhibit both thromboxane A2 formation and arachidonate-induced aggregation in platelet-rich plasma. Despite complete suppression of aggregation with agents that elevate cyclic AMP, considerable thromboxane A2 is still formed. Prostaglandin H2-induced aggregations which bypass the cyclooxygenase regulatory step are also inhibited by agents that elevate cyclic AMP without any measurable effect on thromboxane A2 production. These data demonstrate that cyclic AMP can inhibit platelet aggregation by a mechanism independent of its ability to suppress the cyclooxygenase enzyme. Parallel experiments with washed platelet preparations suggest that they may be an inadequate model for studying the relationship between the platelet cyclooxygenase and platelet function.


Glucocorticoid modulation of prolactin receptors on mammary cells of lactating mice.

Prolactin receptors were monitored by measuring 125I-labeled prolactin binding to collagenase-dissociated mammary epithelial cells of lactating BALB/c mice. Specific receptors for iodine-labeled prolactin with an apparent dissociation constant (Kd) of 0.99 . 10(-9) M were present on the dissociated mammary cells. The binding was inhibited by ovine prolactin, human growth hormone and human placental lactogen but not by follicle stimulating hormone, luteinizing hormone, thyroid stimulating hormone, bovine growth hormone or insulin. Adrenal ablation of nursing mothers caused a reduction of the number of prolactin receptors and this effect was preventable by hydrocortisone therapy. Hydrocortisone injections to mothers 3 days after adrenalectomy also induced a replenishment of the prolactin receptors on the mammary cells. Injections of progesterone failed to sustain the high level of mammary cell prolactin receptors in adrenalectomized animals. Stimultaneous injections of hydrocortisone and progesterone to animals 3 days after adrenalectomy caused a partial suppression of the stimulatory action of hydrocortisone alone. The results suggest that hydrocortisone can exert a modulatory influence on mammary cell prolactin receptors in non-hypophysectomized post-partum mice without altering the dissociation constant (Kd) of the receptors.


Identity of mitochondrial and cytosolic glycerate kinases in rat liver and regulation of their intracellular localization by dietary protein.

Glycerate kinase (ATP: D-glycerate 2-phosphotransferase EC 2.7.1.31) is a key enzyme of glyconeogenesis from serine via hydroxypyruvate. A differential centrifugation of rat liver homogenate and an analysis of the particle fraction by sucrose density gradient centrifugation indicated that 72% and 26% of glycerate kinase are present in mitochondria and cytosol, respectively. A study on the intramitochondrial localization of the enzyme suggested that the mitochondrial glycerate kinase was present in inner membrane and/or matrix. It was found that dietary protein selectively induced mitochondrial glycerate kinase. This result suggested that mitochondrial glycerate kinase had a physiological function for gluconeogenesis from serin. However, the metabolic significance of the cytoplasmic enzyme was still unclear. The properties of solubilized-mitochondrial and cytosolic glycerate kinases were compared. However, no difference between the two enzymes could be found in the kinetic properties, thermal stability, molecular size or electrochemical properties. These results suggested that both enzymes originate from common genetic information. In order to elucidate the regulatory mechanism of the intracellular distribution of glycerate kinase in rat liver, the responses of mitochondrial and cytosolic glycerate kinases to an alteration of dietary protein were studied. The result suggested that an alteration of dietary protein content may regulate the distribution and the translocation of glycerate kinase to mitochondria and cytosol as well as the total amount of glycerate kinase.


Nalpha-trinitrophenyl glucagon: an inhibitor of glucagon-stimulated cyclic AMP production and its effects on glycogenolysis.

Nalpha-Trinitrophenyl glucagon was prepared by reaction with trinitrobenzene sulfonic acid and purified by ion-exchange chromatography. This derivative has essentially no ability to activate adenylate cyclase from rat liver nor to increase the levels of cyclic AMP in isolated hepatocytes nor to stimulate protein kinase activity. This derivative also can act as a glucagon antagonist with regard to cyclic AMP production and can decrease the degree of stimulation of adenylate cyclase caused by glucagon, as well as lowering the glucagon-stimulated elevation of cyclic AMP levels in intact hepatocytes. Nevertheless, this derivative is capable of activating glycogenolysis in isolated hepatocytes and in augmenting the effect of glucagon on glycogenolysis. This metabolic effect of the glucagon derivative thus appears to occur independent of changes in cyclic AMP levels. These results suggest that glucagon can also activate glycogenolysis by a cyclic AM-independent process.


Origin and nature of correlations in the Ia feedback pathway of the muscle control system.

Mammalian primary muscle spindle endings receive two inputs from fusimotor fibres and length changes of their extrafusal environment. Both inputs are also powerful noise sources disturbing the discharge patterns of the receptors. In this paper emphasis is laid on the extrafusal component. It is shown that extrafusal muscle activity can be viewed as a stochastic process which, acting as a common source, can correlate discharge patterns of adjacent muscle spindles. Some quantitative characteristics of these correlations and their relation to the underlying mechanical events are investigated in some detail in order to provide data for more theoretical considerations on their possible physiological importance.


Considerations on mechanisms of focussed signal transmission in the multi-channel muscle stretch reflex system.

This paper presents theoretical considerations on the possibility of topographically ordered signal transmission in the control system of the muscle stretch reflex. It is investigated how correlations between Ia fibres from primary muscle spindle endings in conjunction with an appropriate connectivity of Ia fibres and motoneurones enable the stretch reflex system to trace local routes through the spinal cord. The complex data processing capabilities of the motoneuronal soma-dendritic membrane system are fully taken into account, and it is argued that correlations between inputs to this system may play an important role for signal transmission through the spinal cord.


Application of optimal multichannel filtering to simulated nerve signals.

The optimal linear filters derived in the preceding paper can be thoroughly evaluated using computer simulations, based on the properties of mammalian sensory and motor nerve fibres. Using reasonable values for action potential waveforms, conduction velocity and electrode noise, good separation of motor and sensory signals can be obtained. The performance of the filters is degraded by 1) increasing the electrode noise, 2) introducing dispersion in the conduction velocities, or 3) variation in the waveform of the action potentials from that used in designing the filters. However, the variations needed to seriously degrade performance are quite large compared to those which are likely to be present in mammalian nerves. Use of these filters to distinguish different classes of sensory (or motor) signals based on conduction velocity is discussed.


[Effects of cortisol on lipid and lipoprotein synthesis in rat hepatocytes].

Under in vivo conditions cortisol induces moderate hyperlipidemia followed by an increase in the phospholipid and triglyceride concentrations in the blood and a decrease of cholesterol; similar changes were observed in the liver. At all time intervals studied cortisol inhibits the phospholipid and cholesterol syntheses and decreases the specific radioactivities of the lipids in the mitochondrial fraction. The hormone has an inhibiting effect on the fatty acid synthesis at early postinjection stages. The phospholipid synthesis is increased after adrenalectomy and is then inhibited after injection of the hormone. A single injection of ACTH or cortisol causes suppression of phospholipid and cholesterol syntheses and a decrease in their specific radioactivities in the mitochondria. A similar effect is observed under stress conditions. In addition, the hormone inhibits the synthesis of lipoprotein apoproteins of very low and high densities. After 5 hours following the hormone injection the lipoprotein apoprotein synthesis in the liver is activated; the activation of apoprotein synthesis is also observed after adrenalectomy. However, the injection of the hormone to adrenalectomized rats decreases the apoprotein synthesis. It was shown that in blood serum cortisol affects the conversions of very low density lipoproteins into low density lipoproteins, thus providing for hyperlipidemia.


[Different types of interaction of oxidized and reduced cytochrome c with phospholipid vesicles].

The binding constants of ferri- and ferrocytochrome c interactions with phosphatidylcholine or cardiolipin-containing vesicules were determined. It was found that affinity of ferricytochrome c to phospholipids is one order of magnitude higher that of ferrocytochrome c. A comparative investigation of circular dichroism spectra of free and phospholipid-bound ferri- and ferrocytochrome c was undertaken and it was shown that alpha-helix content of free ferrocytochrome c is higher than that of ferricytochrome c. The formation of ferricytochrome c containing lipoprotein complex led to decrease of alpha-helix content of the protein. In the case of ferrocytochrome c on the other hand interaction with phospholipids did not cause any changes in alpha-helix content. Distribution of ferri- and ferrocytochrome c in different two-phase systems consisting of dextran and polyethylenglycol or dextran and polyethylenglvcol-palmitate was also studied. A comparison of distribution constants shows that higher alpha-helix content of ferrocytochrome c results in the formation of hydrophobic clusters in the protein molecules. In previous communications it was reported that binding of ferrocytochrome c to phospholipids is determined by hydrophobic interactions while in the case of ferricytochrome c the interactions with phospholipids are mainly electrostatic. On the basis of the results obtained in this work it is supposed that it is hydrophobic clusters which determine the binding of ferrocytochrome c to phospholipid membranes.


[Investigation of the effect of phospholipase on Na,K-ATPase activity].

Temperature dependence of bovine brain NA,K-ATPase before and after the short-term treatment of enzyme preparations with phospholipases A, C and D is investigated. Arrhenius plots of the temperature dependence of the reaction rate catalysed by Na,K-ATPase are non-linear, they have an inflection at the region of about 20 degrees C. The treatment of the enzyme with phospholipase A makes the inflection more smooth, phospholipase D shifts the inflection by 4 degrees C to lower temperature and simultaneously activates Na,K-ATPase. Phospholipase C sharply changes the Arrhenius curve and makes it linear. The data obtained are discussed with respect to the role of phospholipids in the formation of membrane bilayer and in the regulation of Na,K-ATPase activity.


[Properties of sex steroid-binding protein from Xenopus laevis blood serum and detection of an estradiol-binding component in frog liver cytosol which differs from sex steroid-binding protein].

Binding and physico-chemical properties of sex steroid-binding protein (SBP) from blood serum and those of estrogen-binding components from liver cytosol of pubertal male and female species of clawed frog Xenopus laevis were studied. It was shown that SBP from both sex species of X. laevis specifically binds estradiol (E2) (Ka=5 . 10(6) M-1). Concentration of SBP binding sites for E2 is 7 . 10(-12) mole per mg of protein. Testosterone 5alpha-dihydrotestosterone and E2 effectively compete with [3H]-E2 for SBP binding sites. Hexestrol, progesterone and corticosterone are weak competitors; estrone and E2-17-hemisuccinate do not compete at all. The Strokes radius of SBP is 4.4 nm; sedimentation coefficient is 4.6S. Molecular weight of SBP is 88000; f/f0 is 1.5 SBP from male frog sera has been purified 8.6-fold with 13% yield. Gel-filtration of [3H]-E2 complexes with liver cytosol proteins shows that the livers of male and female frog X. laevis consol proteins shows that the livers of male and female frog X. laevis contain very low amounts of macromolecular component, which specifically binds E2; this component differs from serum SBP in size and in hormonal specificity. It is assumed that this component is a receptor for estrogens.


Pituitary adrenocortical response to stress during the first day of post-natal life in the rat.

During the first hours of post-natal life, the newborn rat does not respond to ether stress by a rapid increase of ACTH secretion. Ether inhalation by 5- and 12-hour-old animals slightly but significantly increases plasma and adrenal corticosterone levels, but only 30 min after the onset of stress. At 1 day of age, adrenal response is established. The adrenal cortex of 5-hour-old animals responds normally to an intraperitoneal injection of 25 mU of ACTH. These results indicate that there is an absolute stress non-response period in newborn rats which is followed by a relative stress non-response period. The lack of response is not due to an organic block of the adrenal cortex, but could be due to the accumulation of elevated levels of corticosterone in the central nervous system.


Intra-uterine malnutrition and skeletal retardation.

In the subgroup of small-for-dates, indicated as intra-uterine malnourished infants, we made an attempt to correlate the degree of starvation and the degree of skeletal retardation. In 31 wasted infants (including 4 post-mature children) and 20 not wasted infants, the ponderal index (100 X W/L3) was determined and deviation from 'normal' calculated. A highly significant correlation was found between the degree of malnutrition (deviation of the ponderal index from normal) and the degree of skeletal retardation (in weeks). The skeletal retardation seems a useful parameter to determine the minimal duration of insufficient energy supply to the fetus.


Studies in meconium in cystic fibrosis: the activities of alpha-D-mannosidase, beta-glucuronidase, beta-D-fucosidase, acid and alkaline phosphatase.

The specific activities of alpha-D-mannosidase, beta-glucuronidase, beta-D-fucosidase, acid and alkaline phosphatase were studied in meconium from infants with cystic fibrosis (CF) and control subjects. The study revealed significant variations in the specific activity of the enzymes except for acid phosphatase. The variations were not uniform. The activities of alpha-D-mannosidase, beta-glucuronidase and alkaline phosphatase were markedly decreased (p less than 0.001, p less than 0.002, p less than 0.001, respectively), while the activity of beta-D-fucosidase was significantly increased (p less than 0.001) in meconium from the infants with CF. It is suggested that the decreased activity of alpha-D-mannosidase and beta-glucuronidase might contribute to the accumulation of the abnormal substances in CF meconium. The highly increased activity of beta-D-fucosidase raises the possibility of an additional or alternative method for screening newborns for CF using meconium as the test material.


The binding of 125I-TSH to thyroid cell receptors previously deformed (in neonatal age) by gonadotropin treatment.

Single gonadotropin treatment of newborn rats caused a 63-71% decrease of TSH-binding on isolated thyroid cells prepared from thyroid glands of both 6- and 13-month-old animals. This fact points out that: (1) in newborn animals the hormone receptors are still plastic and deformable and (2) the damaging effect of the analogous hormone manifests itself at the receptor level.


The hypotriglyceridemic effect of chenodeoxycholic acid in type IV hyperlipemia.

The effect of chenodeoxycholic acid on the fasting serum triglycerides was studied in 30 patients with type IV hyperlipemia and in 20 patients with primary gout and associated endogenous hypertriglyceridemia, the triglycerides being determined before treatment and at monthly intervals for three months. Chenodeoxycholic acid treatment resulted in a significant lowering of the serum triglycerides in both groups of patients. The drug was well tolerated and there were no undesirable side-effects. Although the mechanism of action is still not known, the drug is thought to reduce triglyceride synthesis in the liver. Chenodeoxycholic acid appears to be electively indicated in type IV hyperlipemia treatment.


In vitro interaction of LDL, anti-lipoprotein IgA and human fibroblasts.

Human LDL were bound and internalized by cultured human fibroblasts. When an antilipoprotein IgA kappa from a case of myeloma with hyperlipidemia and xanthomatosis was introduced into the system, LDL uptake dropped by 50% but LDL binding to fibroblasts was unchanged. In the same system, IgG and IgA controls were inactive.


[Thermal sensitivity of enzymes in trisomy 21 leukocytes].

Thermosensitivity of glucose 6-phosphate dehydrogenase and myloperoxidase was investigated in the leukocyte lysates from patients with Down's syndrome. A statistically significant enhancement of thermosensitivity of both the enzymes investigated was revealed in the aberrant cells in comparison with the control. The noted differences can be based on the increased incidence of mutation injuries in the aberrant cells, as well as on anomalous postranslational modification of the protein molecules.


Hereditary myeloperoxidase deficiency.

The functional properties of granulocytes in a diabetic patient deficient in myeloperoxidase (MPO) were compared with those of granulocytes in healthy subjects. The granulocytes of this patient had normal phagocytic activity. The microbicidal activity of the granulocytes was partially diminished with regard to Staphylococcus aureus and was almost nil with regard to Candida albicans. Fungicidal activity of normal granulocytes was shown to be impaired during the in vitro artificial hyperglycemic condition. The relationship among diabetes mellitus, MPO deficiency, and serious C. albicans infection was examined. Genetic investigation was carried out in 28 members of the proband's family. In close relatives of the patient, MPO values were found to be diminshed to a greater or lesser degree, thus suggesting variable expressivity of the heterozygote state of MPO deficiency.


Action of thrombin on surface glycoproteins of human platelets.

We labeled surface glycoproteins of human platelets by the neuraminidase-galactose oxidase/borotritiide and the periodate/borotritiide methods. When labeled platelets were treated with 1-nM thrombin, a minor glycoprotein weighing 68,000-85,000-d was lost from the surface, and a soluble glycoprotein weighing 57,000-68,000-d was found in the supernatant. Treatment of platelets with ADP, collagen, or the calcium ionophore A23187 did not cause loss of the 68,000-85,000-d glycoprotein from platelet surfaces or appearance of the 57,000-68,000-d glycoprotein in the supernatant. However, trace amounts of the intact 68,000-85,000-d glycoprotein were found in the supernatants of platelets that were not treated with thrombin. The numerous effects of thrombin on platelets could be initiated by cleavage and release the thrombin-sensitive glycoprotein.


In vitro platelet abnormality in adenosine deaminase deficiency and severe combined immunodeficiency.

The platelets of an infant with severe combined immune deficiency and adenosine deaminase deficiency showed markedly diminished responses to ADP-induced aggregation in vitro. This abnormality was corrected by the addition of purified adenosine deaminase in vitro. Exogenous adenosine added to platelet-rich plasma caused markedly prolonged inhibition of ADP-induced aggregation. This was shown by isotopic studies to be due to slow clearance of adenosine and hence persistence of this nucleoside. Direct assay for adenosine deaminiase in plasma and platelet lysates of the patient confirmed the very low activity of this enzyme. Raised cAMP levels were demonstrated in his platelets. The deranged adenosine metabolism and raised cAMP in the platelets of this child with severe combined immunodeficiency may explain the altered response to ADP. Despite the in vitro platelet aggregation abnormality, the patient had no clinical evidence of impaired hemostasis.


Alterations in lysosomes, catalase-containing organelles, mitochondria and plasma membrane fragments from hypertensive rat aorta and caudal artery.

Vascular smooth muscle (VSM) cells from hypertensive and normotensive rat aortae and caudal arteries were isolated by enzymatic techniques, homogenized, and fractionated by differential pelleting. By these techniques, only mitochondria could be enriched more than fivefold in any one fraction. The other organelles were distributed heterogeneously in almost all fractions. Hypertensive smooth muscle enzyme distribution patterns were different from the normotensive, suggesting that changes in sedimentation characteristics had occurred. Activity of the enzyme 5'-nucleotidase increased in whole tissue homogenates and in the 'microsomal' fraction of aortic and caudal artery of hypertensive VSM. The lysosomal protease, cathepsin D, of hypertensive animals decreased in activity for both vascular smooth muscles while N-acetyl-beta-glucosaminidase and pNPPase (acid phosphatase) increased. The possibility of a functional deficiency in protein degradation causing lysosomal overloading is discussed.


Platelet adherence in endothelial cell cultures.

Platelet adherence in bovine endothelial cultures was studied by scanning and transmission electron microscopy. Following incubation with platelet-rich plasma (PRP), platelet adherence to endothelial cell surfaces was rare as compared to similarly-tested fibroblasts which displayed numerous adherent platelets. When endothelial cells were induced to retract from their substrate by exposure to cold or versene and then incubated with PRP, platelets were observed adhering to endothelial cell processes and to an extracellular microfilamentous network, located beneath the cell. Platelets were attached singly, retained their discoidal shape, and showed no evidence of granule release. In contrast, microfilaments and adherent platelets were conspicuously absent in endothelial cultures which were retracted with trypsin or collagenase and incubated with PRP. These preliminary results suggest that the observed interaction between platelets and the subcellular surface of cultured endothelial cells is specific for an extracellular network of microfilaments produced by the cells.


Acupuncture in narcotic withdrawal: a preliminary report on biochemical changes in the blood and urine of heroin addicts.

Eight female heroin-addicted subjects were treated over a 10-day period by acupuncture and electrical stimulation (AES). Their plasma ACTH, cortisol and c-AMP levels were reduced after initial AES treatment. The reduction of c-AMP was significant the most of the three. The suppression of these compounds may be associated with the temporary relief of withdrawl symptoms on and after each AES treatment in the first three days. On the third day, ACTH, cortisol and c-AMP levels did not show significant changes after AES. This was presumably due to the disappearance of withdrawal symptoms and possibly to the stabilizing effect of the treatment. Both the ACTH and c-AMP levels were generally lower at the end of the treatment period. Results of the present investigation suggest that plasma c-AMP may be the best parameter by which to gauge the response of heroin addicts to AES.


The effects of opiate receptor agonists and antagonists on the stress-induced secretion of corticosterone in mice.

1 Intraperitoneal administration of normorphine, morphine or naloxone or exposure to ether vapour for 1 min, elevated plasma corticosteroid concentrations in mice. 2 Injection of saline or exposure to ether vapour rendered mice less sensitive to a subsequent exposure to ether vapour 15 min later. 3 Treatment with normorphine (50 mg/kg) potentiated the corticosteroid response to ether stress whilst pentazocine (20 mg/kg), naltrexone (10 mg/kg), morphine (24 mg/kg), levorphanol (20 mg/kg) and naloxone (50 mg/kg) prevented the stress-induced elevation of plasma corticosteroids. 4 Both naloxone and morphine inhibited the potentiation by normorphine of the response to ether, the dose of naloxone required being higher than that for inhibition of normorphine analgesia. 5 It is concluded that endogenous opioid peptides may be involved in the control of the response to ether stress in mice.