Segregation of FMRF amide-immunoreactive efferent fibers from NPY-immunoreactive amacrine cells in goldfish retina

Summary Immunocytochemical studies were conducted on goldfish to determine whether a retinal efferent fiber system, immunoreactive to the tetrapeptide Phe-Met-Arg-Phe-NH₂ (FMRFamide), might contain instead a substance similar to one of the 36-amino acid pancreatic polypeptides, the C-terminus of which is similar to FMRFamide.Our results demonstrate the presence of two separate peptidergic systems, one containing FMRFamide-like, and the other pancreatic polypeptide-like peptides. Antisera to FMRFamide reveal the efferent fibers, whose axons exit the optic nerve and terminate in layer 1 of the inner plexiform layer, as previously described. Antisera to porcine neuropeptide Y, and to avian and bovine pancreatic polypeptides label a sparse population of putative amacrine cell bodies and a dense fiber plexus in layers 1, 3, and 5 of the inner plexiform layer. Based on intensity of staining, this amacrine cell peptide appears to be most similar to neuropeptide-Y.Radioimmunoassay and immunocytochemical staining of retinas in which the efferent fiber peptide was depleted by optic nerve crush confirm in large part the observation that the two peptide systems are distinct. However, there is some cross-recognition of the FMRFamide-like tissue antigen by pancreatic polypeptide antibodies.Double-label studies with antisera to tyrosine hydroxylase and neuropeptide-Y indicate that the pancreatic polypeptide antigen is not co-localized with catecholamines.     

Acetoacetyl Coenzyme A Reductase and Polyhydroxybutyrate Synthesis in Rhizobium (Cicer) sp. Strain CC 1192

Biochemical controls that regulate the biosynthesis of poly-3-hydroxybutyrate (PHB) were investigated in Rhizobium (Cicer) sp. strain CC 1192. This species is of interest for studying PHB synthesis because the polymer accumulates to a large extent in free-living cells but not in bacteroids during nitrogen-fixing symbiosis with chickpea (Cicer arietinum L.) plants. Evidence is presented that indicates that CC 1192 cells retain the enzymic capacity to synthesize PHB when they differentiate from the free-living state to the bacteroid state. This evidence includes the incorporation by CC 1192 bacteroids of radiolabel from [¹⁴C]malate into 3-hydroxybutyrate which was derived by chemically degrading insoluble material from bacteroid pellets. Furthermore, the presence of an NADPH-dependent acetoacetyl coenzyme A (CoA) reductase, which was specific for R-(−)-3-hydroxybutyryl-CoA and NADP⁺ in the oxidative direction, was demonstrated in extracts from free-living and bacteroid cells of CC 1192. Activity of this enzyme in the reductive direction appeared to be regulated at the biochemical level mainly by the availability of substrates. The CC 1192 cells also contained an NADH-specific acetoacetyl-CoA reductase which oxidized S-(+)-3-hydroxybutyryl-CoA. A membrane preparation from CC 1192 bacteroids readily oxidized NADH but not NADPH, which is suggested to be a major source of reductant for nitrogenase. Thus, a high ratio of NADPH to NADP⁺, which could enhance delivery of reductant to nitrogenase, could also favor the reduction of acetoacetyl-CoA for PHB synthesis. This would mean that fine controls that regulate the partitioning of acetyl-CoA between citrate synthase and 3-ketothiolase are important in determining whether PHB accumulates.     

Effects of high altitude stay on the incidence of common diseases in man

Prolonged stay at high altitude significantly lowers the incidence of some of the diseases commonly encountered at sea level. This conclusion is based on a study involving 130,700 men stationed on plains between 760 m and sea level and 20,000 men stationed at altitudes between 3692 and 5538 m during the period 1965 to 1972. When yearwise differences in morbidity rates were determined for this period it was found that apart from amoebic hepatitis, goitre and lobar pneumonia, which show a higher incidence, the incidence of infections of bacterial, viral and protozoal origin, diabetes mellitus, hypertension and ischaemic heart disease, asthma and rheumatoid arthritis, gastric disorders, skin diseases, psychiatric ailments and anaemia was significantly lower at high altitude than at sea level. When the trend in morbidity rates was compared over the two subperiods of 1965 to 1968 and 1969 to 1972 it was found that generally increasing or decreasing trends on plains were reflected at high altitude. The overall incidence at high altitude however remained low. Part I of our communication deals with epidemiological data and these findings. Part II surveys the available literature and attempts to explain how improved hormonal state, enhanced fibrinolytic activity, accelerated humoral and cellular immune responses, favourable haemodynamics, better cardiac and cerebral functions, improved metabolic functions, and a relatively stable, dry and cold climate favourably influence the incidence of diseases at high altitude.     

Influence of Frequently Used Chemical Insecticides on Mycoflora Carried by Common Housefly, <Emphasis Type="Italic">Musca domestica</Emphasis> L.

The housefly, Musca domestica L. (Diptera: Muscidae), is a major medical and veterinary insect pest. It serves as a vector of many pathogenic microorganisms causing spoilage of food and diseases in human and animals. Use of chemical insecticides is adapted as a principal tool to manage housefly. Insecticides have many unforeseen ecological consequences including effects on non-target organisms. In the present study, we have assessed the effects of 10 different synthetic insecticides on the growth of mycoflora associated with the external body of the housefly by using poison food technique. Our results reveled that all synthetic insecticides enhanced the growth. Surprisingly, in most of the cases, mycelial growth of fungi was significantly increased at high concentration as compared with lower concentration. This study provides useful information about the dangerous effects of synthetic insecticides on environment by increasing the spread of various non-target pathogenic, mycotoxigenic, and food spoiling fungi, carried by houseflies.     

Diversity in virus populations from genital secretions and peripheral blood from women recently infected with human immunodeficiency virus type 1.

In order to develop a human immunodeficiency virus type 1 vaccine with global efficacy, it is important to evaluate the virus populations that are transmitted to individuals living in high-incidence areas. To determine the nature of the human immunodeficiency virus type 1 population transmitted to women during heterosexual contact, we examined the diversity of the proviral envelope gene in infected cells in both genital secretions and peripheral blood from six recently seroconverted Kenyan women. Heterogeneous virus populations were present in cervical secretions and/or peripheral blood shortly after seroconversion for five of six infected individuals, and tissue-specific variants were identified in several cases.     

Reversal of abnormal fibrinolytic activity,blood coagulation factors and platelet function in high-altitude pulmonary oedema with frusemide

In high-altitude pulmonary oedema there is evidence of widespread intravascular clotting which is associated with and may result from reduced fibrinolytic activity with increased plasma fibrinogen, increased factors V, VIII and X, decreased factor XII, and increased platelet adhesiveness and platelet factor 3. The beneficial effect of frusemide which is the mainstay of treatment has so far been ascribed to its potency and rapidity of action in inducing diuresis and its effect on pulmonary blood volume which is reduced. However, an interesting new finding is that at the same time frusemide reverses adverse changes in fibrinolytic activity, blood coagulation, and platelet function and thereby removes impediment to the pulmonary blood flow at the capillary and venular level.

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Zusammenfassung Bei Lungenödem in Hochgebirge liegt eine weitverteilte, intravasale Gerinnung vor, die verbunden ist mit oder als Folge einer verminderten fibrinolytischen Aktivität mit erhöhtem Plasmafibrinogen, Faktoren V, VIII und X und vermindertem Faktor XII, erhöhter Plättchen-Klebrigkeit und Plättchenfaktor 3 auftritt. Die günstige Wirkung von Frusemid wurde bisher seiner raschen Wirkung auf die Auslösung der Diurese und der Reduzierung des pulmonalen Blutvolumens zugeschrieben. Nach einer neuen Beobachtung kehrt Frusemid die Veränderungen der fibrinolytischen Aktivität, Blutgerinnung und Plättchenfunktion um und beseitigt auf diese Weise die Behinderung des pulmonalen Blutflusses im Kapillarbereich.

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Resume Les oedèmes pulmonaires constatés en haute montagne sont en étroite relation avec une augmentation des facteurs de coagulation du sang. Cette augmentation est consécutive à une diminution de l'activité fibrinolytique. Elle est accompagnée d'une augmentation des fibrines du plasma, des facteurs V, VIII et X et d'une diminution du facteur XII. On constate en outre une adhésion plus forte des plaquettes et une hausse du facteur 3 des dites plaquettes. On a attribué l'action bienfaisante de la frusémide au fait qu'elle déclenche rapidement une diurèse et une réduction du volume du sang dans les poumons. Cependant, de nouvelles observations ont amené à la constatation que la frusémide renverse les processus de modification de l'activité fibrinolytique, de la coagulabilité du sang et des fonctions des plaquettes. En outre, elle écarte de cette façon la rétention du flux sanguin pulmonaire dans la zone des capillaires.

     

Differential coupling through Val-344 and Tyr-442 of trimethylamine dehydrogenase in electron transfer reactions with ferricenium ions and electron transferring flavoprotein

Modeling studies of the trimethylamine dehydrogenase-electron transferring flavoprotein (TMADH-ETF) electron transfer complex have suggested potential roles for Val-344 and Tyr-442, found on the surface of TMADH, in electronic coupling between the 4Fe-4S center of TMADH and the FAD of ETF. The importance of these residues in electron transfer, both to ETF and to the artificial electron acceptor, ferricenium (Fc(+)), has been studied by site-directed mutagenesis and stopped-flow spectroscopy. Reduction of the 6-(S)-cysteinyl FMN in TMADH is not affected by mutation of either Tyr-442 or Val-344 to a variety of alternate side chains, although there are modest changes in the rate of internal electron transfer from the 6-(S)-cysteinyl FMN to the 4Fe-4S center. The kinetics of electron transfer from the 4Fe-4S center to Fc(+) are sensitive to mutations at position 344. The introduction of smaller side chains (Ala-344, Cys-344, and Gly-344) leads to enhanced rates of electron transfer, and likely reflects shortened electron transfer "pathways" from the 4Fe-4S center to Fc(+). The introduction of larger side chains (Ile-344 and Tyr-344) reduces substantially the rate of electron transfer to Fc(+). Electron transfer to ETF is not affected, to any large extent, by mutation of Val-344. In contrast, mutation of Tyr-442 to Phe, Leu, Cys, and Gly leads to major reductions in the rate of electron transfer to ETF, but not to Fc(+). The data indicate that electron transfer to Fc(+) is via the shortest pathway from the 4Fe-4S center of TMADH to the surface of the enzyme. Val-344 is located at the end of this pathway at the bottom of a small groove on the surface of TMADH, and Fc(+) can penetrate this groove to facilitate good electronic coupling with the 4Fe-4S center. With ETF as an electron acceptor, the observed rate of electron transfer is substantially reduced on mutation of Tyr-442, but not Val-344. We conclude that the flavin of ETF does not penetrate fully the groove on the surface of TMADH, and that electron transfer from the 4Fe-4S center to ETF may involve a longer pathway involving Tyr-442. Mutation of Tyr-442 likely disrupts electron transfer by perturbing the interaction geometry of TMADH and ETF in the productive electron transfer complex, leading to less efficient coupling between the redox centers.     

The ATPase cycle of Hsp90 drives a molecular 'clamp' via transient dimerization of the N-terminal domains

How the ATPase activity of Heat shock protein 90 (Hsp90) is coupled to client protein activation remains obscure. Using truncation and missense mutants of Hsp90, we analysed the structural implications of its ATPase cycle. C-terminal truncation mutants lacking inherent dimerization displayed reduced ATPase activity, but dimerized in the presence of 5'-adenylamido-diphosphate (AMP-PNP), and AMP-PNP- promoted association of N-termini in intact Hsp90 dimers was demonstrated. Recruitment of p23/Sba1 to C-terminal truncation mutants also required AMP-PNP-dependent dimerization. The temperature- sensitive (ts) mutant T101I had normal ATP affinity but reduced ATPase activity and AMP-PNP-dependent N-terminal association, whereas the ts mutant T22I displayed enhanced ATPase activity and AMP-PNP-dependent N-terminal dimerization, indicating a close correlation between these properties. The locations of these residues suggest that the conformation of the 'lid' segment (residues 100-121) couples ATP binding to N-terminal association. Consistent with this, a mutation designed to favour 'lid' closure (A107N) substantially enhanced ATPase activity and N-terminal dimerization. These data show that Hsp90 has a molecular 'clamp' mechanism, similar to DNA gyrase and MutL, whose opening and closing by transient N-terminal dimerization are directly coupled to the ATPase cycle.