Effect of alpha-MSH on the corticosteroid production of isolated zona glomerulosa and zona fasciculata cells

α-MSH (10^?9 ? 6×10^?7M) potentiates the effect of ACTH (10^?11 ? 5×10^?9M) on adrenocortical steroidogenesis decreassng ED₅₀ of ACTH from 220 to 183 pg/ml on zona fasciculata corticosterone-, and from 739 to 437 pg/ml on zona glomerulosa aldosterone production. α-MSH alone increases aldosterone production of zona glomerulosa cells in doses (10^?9 ? 6×10^?7M) that do not stimulate zona fasciculata corticosterone production. The response of zona glomerulosa aldosterone production to α-MSH can be characterized by a bi-phase dose-response curve.     

Low prostaglandin concentrations cause cardiac rhythm disturbances. Effect reversed by low levels of copper or chloroquine

IN perfused male rat hearts concentrations of prostaglandins (PGs) E2 and F2α in the range 1 pg/ml to 10 ng/ml (2.8 × 10⁻¹² to 2.8 × 10⁻⁸M) consistently caused rhythm irregularities. Higher concentrations had no effect themselves and stabilized rhythm in hearts made unstable by lower concentrations. Copper ions (as the sulphate) at 2 × 10⁻⁶M stabilized hearts made unstable by PGs and when present prior to the PGs prevented PG induced disturbances. Chloroquine also reversed PG-induced rhythm changes.     

The relationship between sperm concentration and fertilization in vitro of rabbit ova

Different concentrations of in utero incubated rabbit sperm (1.5 × 10⁴-120 × 10⁴ /ml) were tested to determine whether there is a relationship between sperm concentration and level of fertilization achieved “in vitro” of rabbit ova. While low concentrations (1.5 × 10⁴-4.5 × 10⁴ /ml) resulted in relatively low fertilization (23–36%), those in the range of 13 × 10⁴?120 × 10⁴ /ml gave fertilization rates of 65–83%. Consistently high results were obtained with sperm counts above 40 × 10⁴ /ml. This is in agreement with the concentration of spermatozoa found in vivo in the Fallopian tubes around the time of fertilization (50 × 10⁴ /ml).     

Chemiluminescence properties of human,canine and rat polymorphonuclear cells

Human as well as canine and rat polymorphonuclear cells (PMN) were separated from whole blood by centrifugation. Two-step discontinuous Percoll gradients with distinct different densities were used. The chemiluminescence properties of the isolated PMN and of phagocytes in small quantities of whole blood were compared in luminol-enhanced assays after stimulation with various agents: non-opsonized zymosan (3.5 g/I), phorbol myristate acetate (PMA, 2.8 × 10^?6 mol/I), calcium ionophore A 23187 (10^?5 mol/l) and N-formylmethionyl-leucyl-phenylalanine (FMLP, 3.5 × 10^?6 mol/l). The isolated cells of the three species responded to all of the various stimuli. Species-related sensitivity could be ordered: human > canine > rat. Response to the various agents in the human cells can be ranked: PMA ? A 23187 > zymosan > FMLP; for the dog: A 23187 > PMA > zymosan > FMLP; and for the rat: zymosan ? PMA > FMLP ? A 23187. Time course and peak maximum response were different upon stimulation in the absence and presence of autologous plasma. Distinct soluble stimuli resulted in maximum responses below the baseline in the whole blood assays with canine (FMLP) and rat (FMLP, A 23187) phagocytes.     

Hydrochemical,bacteriological assessment,and classification of groundwater quality in Thulamela Municipality,South Africa: potential health risk

Abstract

Potential effects of anthropogenic activities on different boreholes around Thulamela Municipality, South Africa, were evaluated by quantifying the bacteria indicators and physicochemical parameters during summer, autumn, and winter. The purpose was to determine whether the borehole water in this region is safe for domestic use across the seasons. The concentrations of physicochemical (Temperature, pH, electrical conductivity (EC), turbidity, and nitrate) and bacteriological (both Escherichia coli and Enterococcus faecalis) contaminants in the borehole water samples were determined using standard microbiology methods. The mean concentration of NO₃ for most of the boreholes failed to comply with the recommended guidelines throughout the season. High microbial load of E. coli (2.0?×?10¹ – 4.6?×?10³ CFU/100?ml) and E. faecalis (2.0?×?10° – 6.0?×?10² CFU/100?ml) was recorded in the wet season than in the dry season (0.0 – 7.0?×?10² and 0.0 – 1.0?×?10¹ CFU/100?ml, respectively). Sanitary inspection and water source classification showed that most boreholes are prone to chemical contaminants during summer and autumn due to rainfall and this correlates with the measured microbial contamination. The result is significantly important because water from these boreholes is used for domestic purposes without treatment and could pose major public health risks to the consumers.     

Glycolysis and glucose oxidation during reperfusion of ischemic hearts from diabetic rats

Stimulationg of glucose oxidation by dichloroacetate (DCA) treatment is beneficial during recovery of ischemic hearts from non-diabetic rats. We therfore determined whether DCA treatment of diabetic rat hearts (in which glucose use is extremely low), increases recovery of function of hearts reperfused following ischemia. Isolated working hearts from 6 week streptozotocindiabetic rats were perfused with 11 mM [2-³H/U-¹⁴C]glucose, 1.2 mM palmitate, 20 μU/ml insulin, and subjected to 30 min of no flow ischemia followed by 60 min reperfusion. Heart function (expressed as the product of heart rate and peak systolic pressure), prior to ischemia, was depressed in diabetic hearts compared to controls (HR × PSP × 10^?3 was 18.2 ± 1 and 24.3 ± 1 beats/mm Hg/min in diabetic and control hearts respectively) but recover to pre-ischemic levels following ischemia, whereas recovery of control of control hearts was significantly decreased (17.8 ± 1 and 11.9 ± 3 beats/mm Hg/min in diabetic and control hearts respectively). This enhanced recovery of diabetic rat hearts occurred even though glucose oxidation during reperfusion was significantly reduced as compared to controls (39 ± 6 and 208 ± 42 nmol/min/g dry wt, in diabetic and control hearts respectively). Glycolytic rate (³G₂O production) during reperfusion were similar in diabetic and control hearts (1623 ± 359 and 2071 ± 288 nmol/min/g dry wt, respectively). If DCA (1 mM) was added at reperfusion, hearts from control animals exhibited a significant improvement in function (HR × PSP × 10^? recovered to 20 ± 4 beats/mm Hg/min) that was accompanied by a 4-fold increase in glucose oxidation (from 208 ± 42 to 753 ± 111 nmol/min/g dry wt). DCA was without effect on functional recovery of diabetic rat hearts during reperfusion but did significantly increase glucose oxidation from 39 ± 6 to 179 ± 44 nmol/min/g dry wt). These data suggests that, unlike control hearts, low glucose oxidation rates are not an important factor in reperfusion recovery of previouskly ischemic diabetic rat hearts.     

Isolation of eburnetoxin,A vasoactive substance from the Conus eburneus venom

Eburnetoxin, a powerful vasoactive protein has been isolated from the venom of the marine snail Conus eburneus, monitored by the contractile effect to the rabbit aorta. The molecular weight was estimated to be 28, 000 by gel permeation chromatography and slab gel electrophoresis. The purified protein was electrophoretically homogeneous. The toxin at concentrations above 3 × 10^?7 g/ml elicited a marked contractile response of aorta, which was inhibited by verapamil (10^?6 M). The minimum lethal dose in the fish Rhodeus ocellatus smithi was 1 μg/g body weight.     

Epidermal growth factor: Characteristics of specific binding in membranes from liver,placenta, and other target tissues

Epidermal growth factor, a 6,400-dalton polypeptide from the mouse submaxillary gland, binds specifically to cells and membranes derived from a variety of human, rat, mouse, and bovine tissues. Liver, placenta, skin, cornea, and cultured chondrocytes, Hela cells, and Chang liver cells bind large amounts of epidermal growth factor, whereas fat cells, resting and lectin-stimulated human peripheral lymphocytes, mouse thymocytes, cultured rat hepatoma cells, and mammary cells from virgin and pregnant mice bind little or no epidermal growth factor. The binding site for epidermal growth factor is distinct from receptors for other anabolic peptides such as insulin, nerve growth factor, and growth hormone. The binding of epidermal growth factor is rapid and reversible. The rate constant of association is approximately 10⁶ mole^?1 sec^?1, the rate constant of dissociation is about 6 × 10^?4 sec^?1, and the apparent equilibrium dissociation constant is about 10^?9m. Trypsin at low concentrations (50–200 μg/ml) destroys the receptor site for epidermal growth factor. The binding of epidermal growth factor by membranes is not accompanied by appreciable degradation of the peptide present in the medium or of that bound to the membranes. Use can be made of the high affinity and specificity of membranes for epidermal growth factor to measure by a competitive binding assay as little as 200 pg of EGF per ml (3 × 10^?11m).     

The Effect of Manganese-induced Cytotoxicity on mRNA Expressions of HSP27, HSP40, HSP60, HSP70 and HSP90 in Chicken Spleen Lymphocytes in Vitro

The purpose of this study was to investigate the effect of manganese (Mn)-induced cytotoxicity on heat shock proteins in chicken spleen lymphocytes. Lymphocytes were cultured in medium in the absence and presence of MnCl₂ (2?×?10^?4, 4?×?10^?4, 6?×?10^?4, 8?×?10^?4, 10?×?10^?4, and 12?×?10^?4 mmol/L) for 12, 24, 36, and 48 h in vitro. Then, the mRNA levels of HSP27, HSP40, HSP60, HSP70, and HSP90 were examined by real-time quantitative PCR. The results showed that the mRNA levels of HSP27, HSP40, HSP60, HSP70, and HSP90 in all treatment groups at all time points, except mRNA levels of HSP27 at 48 h, had the same tendency. As manganese concentration increased, the mRNA expression of the heat shock proteins first increased and then decreased. In other words, we demonstrated that the mRNA expression of the heat shock proteins was induced at lower concentrations of manganese and was inhibited at higher concentrations. Mn had a dosage-dependent effect on HSP27, HSP40, HSP60, HSP70, and HSP90 mRNA expression in chicken spleen lymphocytes in vitro.     

Rat atrial natriuretic factor: Isolation,structure and biological activities of four major peptides

Four peptides possessing both natriuretic activity and smooth muscle relaxant activity were isolated from rat atrium and their amino acid sequences determined. The four peptides designated ANF-I, ANF-II, ANF-III and ANF-IV containing 35, 31, 30 and 25 amino acid residues, respectively, were obtained in a molar ratio of 4:60:20:16. The predominant species ANF-II, which may represent the native form of ANF, has the following sequence: (H₂N)-G-P-R-S-L-R-R-S-S-C-F-G-G-R-I-D-R-I-G-A-Q-S-G-L-G-C-N-S-F-R-Y-(COOH) in which Cys-10 and Cys-26 are disulfide linked. Cleavage of the aspartyl linkage at position 16 by staphylococcal protease caused complete inactivation, indicating that the ring conformation is essential. The dose-response relationships determined for the four pepdides in relaxing norepinephrine-induced contraction of rabbit thoracic aorta showed half-maximum relaxation at concentrations ranging from 1.5 × 10^?9 to 2.5 × 10^?9 M. Comparable dose-response relationships were observed in relaxation of carbacol-induced contraction of chick rectum strips as tested with ANF-II and ANF-IV.     

Myocardial actions of prostaglandins in isolated cat cardiac tissue.

The inotropic responses to prostaglandins (PG) A₁, E₁, E₂ and F_2α were studied in isolated cat myocardial tissue. PGA₁ and F_2α exhibited no significant inotropic effects, whereas, PGE₂ and PGE₁ produced negative inotropic effects at concentrations of 2.8 × 10⁻⁷ and 2.8 × 10⁻⁶ M in isolated cat papillary muscles.In isolated perfused cat hearts, PGE₁ (2.8 × 10⁻⁶M) produced a negative inotropic effect along with a significant increase in coronary flow. As flow declined, the negative inotropic effect became more severe. PGE₁ at 2.8 × 10⁻⁹ M produced a sustained increase in coronary flow and oxygen consumption with no inotropic effect. PGE₂ and F_2α did not exert significant changes in coronary flow or contractile force.Thus prostaglandins do not appear to exert significant positive inotropic effects at physiologic or at generally accepted pharmacologic concentrations in isolated cat heart preparations. At extremely high concentrations, prostaglandins E₁ and E₂ exert a negative inotropic effect; however, this would not explain the protective effect of these prostaglandins in circulatory shock.     

Control of green mould of citrus by using Trichoderma harzianum,humic acid or garlic

Penicillium digitatum, an aggressive fungus causes post-harvest decay of mandarin sweet orange and Washington navel. In vitro Trichoderma harzianum or humic acid (HA) or powdered cloves of garlic caused inhibition of fungal growth of isolates P₁ and P₂. Under storage conditions, the fruit citrus is protected by using T. harzianum with standard volume 2.0?ml (9.6?×?10⁶?conidia/ml) and application 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum spore suspension (1.0?×?10⁶?spores/ml) compared to control. Spraying the fruit citrus by standard volume of 2.0?ml of either HA or powder cloves of garlic 1% on each fruit 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum (1.0?×?10⁶ spores/ml) compared to control. The lowest percentage of disease incidence and disease severity were associated with powder of cloves garlic and followed by HA and T. harzianum during two growing seasons compared with the untreated and control.     

Distribution,sources, and health risk assessment of polycyclic aromatic hydrocarbons in dust from urban environment in the Niger Delta,Nigeria

The concentrations of the 16 U.S. Environmental Protection Agency polycyclic aromatic hydrocarbons (PAHs) were measured in urban street dust with a view to understanding the potential risk to urban residents exposed to street dust, sources, and distribution of the PAHs. Gas chromatography–mass spectrometry was used to measure the concentrations of the PAHs after ultra-sonication with hexane/dichloromethane and clean up. The total concentrations of the ?16 PAHs in the urban street dust from Warri and its environs ranged from 165.1 to 1012 µg kg^?1. The isomeric ratios and the PCA indicated that combustion of petroleum and biomass are the major sources of PAHs in Warri and its environs. The total BaP carcinogenic and mutagenic potency concentrations (?BaP_TEQ and ?BaP_MEQ) were in the range of 0.03 to 219 µg kg^?1 and 0.52 to 182 µg kg^?1, respectively. The incremental life cancer risk from exposure of the residents of Warri and its environs ranged from 4.07 × 10^?7 to 3.11 × 10^?3 and 2.13 × 10^?7 to 1.49 × 10^?3 for the children and adults, respectively, which were higher than the baseline value of acceptable risk of 10^?6 (i.e., one case per million people).     

Development of an experimental laboratory fertilization protocol for the South African abalone,Haliotis midae (Linnaeus 1758)

In this study, effective gamete concentrations, egg viability, and fertilization volumes were evaluated for Haliotis midae (L.). Sperm concentrations between 5?×?10³ and 5?×?10⁴?mL^?1 (p?>?0.05) consistently resulted in high hatch-out rates (96?±?1%). At concentrations higher than 5?×?10⁵?mL^?1, hatch-out rates decreased to 69?±?7% (p??1 resulted in high fertilization rates, with 50?eggs?mL^?1 being the ideal concentration for fertilization in H. midae. Egg viability was consistently high up to 100?min post-spawning, with a decrease in hatch-out success, when eggs were fertilized 120?min post-spawning. Fertilization volumes did not affect successful hatch-out. The results from this study can be implemented by South African abalone farms to increase hatch-out rates and subsequent culture. It can also be used as basis for the development of fertilization protocols in other marine invertebrate species.     

Effects of prostacyclin on the canine isolated basilar artery

Prostacyclin (PGI₂) produced a biphasic response in canine isolated basilar arteries. In low doses (1 × 10^?8M?1 × 10^?7M) PGI₂ caused a slight but consistent relaxation of resting muscle tone. In low concentrations (1 × 10^?8M?1 × 10^?6M) PGI₂ antagonized muscle contractions caused by serotonin or prostaglandin (PG) F_2α. This relaxant effect with low doses of PGI₂ on the isolated cerebral artery contrasts with findings obtained with other PGs and supports the hypothesis that PGI₂ is a mediator of vasodilatation. However, in 1 × 10^?5M concentrations PGI₂ contracted the arterial muscle and did not antagonize contractions induced by serotonin or PGF_2α.     

Effect of the cations sodium,potassium and calcium on the interaction of hyaluronate chains: a light scattering and viscometric study

Light scattering and viscometric studies have been carried out on two preparations, A and B, of rooster comb hyaluronate. Sedimentation rate studies have also been performed with A. Light scattering measurements in 0.2 m KCl for preparation A gave a molecular weight of 3.3 × 10⁶ and for B, 1.0 × 10⁶. In (0.1–0.3) M NaCl similar measurements gave a particle weight for A of (4.4–6.4 × 10⁶ and for B (1.7–2.8 × 10⁶. In 0.066 m CaCl₂ molecular weight values of 9.5 × 10⁶ for A and 1.7 × 10⁶ for B were obtained. Thus in the presence of Na⁺ and Ca²⁺ ions aggregates of chains persisted into dilute solution. Measurements by light scattering on A and B in 4 m guanidinium chloride gave values in the same range as those obtained in 0.2 m KCl. Sedimentation rate studies on A gave values of 10.3 Svedbergs in 0.2 m KCl and 12.2 Svedbergs in 0.2 m NaCl and 0.066m CaCl₂. The shear dependence of the viscosity was studied using a conicylindrical viscometer at shear rates between 0.5 and 20 s^?1. Preparation A in 0.2 m KCl and NaCl yielded values for (ν_sp/cc→0 of 5000 and 7100 ml g^?1 respectively in keeping with the tendency to aggregate. The behaviour for preparation B was similar. In 0.066 m CaCl₂ there was a marked dependence of viscosity on shear speed below 10 s^?1 for all concentrations and the value of (ν_sp/c)→0 at 0 s^?1 for preparation A was 7700 ml g^?1 while at a shear rate of 8 s^?1 (ν_sp/c)c→0 ? 5000 ml g ^?1. Similar effects were found for preparation B and the data suggest associations of chains disruptable by weak shear forces. The increase in viscosity with concentration in the presence of 0.066 m CaCl₂ was much less than in the presence of KCl or NaCl, suggesting that the Ca²⁺ had a marked effect on the ”rigidity’ of the molecules in solution. A viscometric titration experiment with Ca^2? showed that a level of 0.02 m CaCl₂ in 0.2 m NaCl was sufficient to produce the change in viscosity presented above and that significant perturbations of the viscosity were present at 0.005?0.01 m CaCl₂.     

Identification of a bacterium isolated from the diseased brown planthopper and determination of its insecticidal activity

The brown planthopper, Nilaparvata lugens, is one of the most harmful insect pests of rice crops in Asian countries. To find an effective biological control agent against this pest, we investigated the bacterial flora of field N. lugens collected from Jiangsu Province, China, in 2012 and tested its insecticidal activity. A novel bacterium strain, S-JS1, was isolated from N. lugens nymphs and adults and showed a high level of insecticidal activity. Based on its phenotypic, physiological and biochemical properties, and its 16S rRNA gene phylogeny, the isolate was assigned to Serratia marcescens; the name S. marcescens S-JS1 is proposed. The pathogenicity of S-JS1 against the third-instar nymphs, and the macropterous and the brachypterous adults of N. lugens were compared. The median lethal concentration (LC₅₀) values of S-JS1 against the brachypterous adult were the lowest (LC₅₀, 1.53?×?10⁸ colony forming units (cfu)/ml), followed against the macropterous adult (LC₅₀, 1.65?×?10⁹?cfu/ml) and third-instar nymphs (LC₅₀, 1.86?×?10⁹?cfu/ml) at 5 days post-infection. The median lethal time values of 8?×?10⁸?cfu/ml S. marcescens S-JS1 against the brachypterous adult, macropterous adult, and third-instar nymph were 4.5, 5.5, and 5.7 days, respectively. These results indicate that the S-JS1 isolate appears to be a promising S. marcescens strain with strong biocontrol potential against N. lugens.     

Introduction of Beauveria bassiana as a biological control agent against the Ommatissus lybicus in Kerman province

Dubas bug, Ommatissus lybicus is one of the major sucking pests of date palm. Data palm is the most important economic crop in Iran. In this research, we isolated the pathogenic strain KB 201, KB 206, KB 211, KB 215, KB 220 and KB 512, of entomopathogenic fungi Beauveria bassiana from soil and insects, which produced aerial and submerged conidia and blastospores in the laboratory conditions. We investigated the best conditions for the production and utilisation of the spore suspension to spray the nymph of O. lybicus, which is one of the important pests in Kerman province (Iran). A total of 180 isolates that naturally infected by O. lybicus were collected from date palm in Kerman province during spring and summer seasons 2010–2011. The pathogen city test was carried out with direct spray. To bioassay the isolates, three concentrations of the spore suspension were prepared as follows: 1?×?10⁶, 1?×?10⁷ and 1?×?10⁸?conidia/ml. The pests were sprayed by aerial conidial suspension, which was prepared by 0.01% Tween 80 in distilled water, and the controls were sprayed by 0.01% Tween 80in distilled water. After spraying the pests, the plates were incubated at 25?±?1?°C and 80% of relative humidity. Then, the treated pests were monitored every day for the fungal growth and mortality.     

Diversity, abundance, and activity of ammonia-oxidizing bacteria and archaea in Chongming eastern intertidal sediments

Ammonia oxidation plays a pivotal role in the cycling and removal of nitrogen in aquatic sediments. Certain bacterial groups and a novel group of archaea, which is affiliated with the novel phylum Thaumarchaeota, can perform this initial nitrification step. We examined the diversity and abundance of ammonia-oxidizing β-Proteobacteria (β-AOB) and ammonia-oxidizing archaea (AOA) in the sediments of Chongming eastern tidal flat using the ammonia monooxygenase-α subunit (amoA) gene as functional markers. Clone library analysis showed that AOA had a higher diversity of amoA gene than β-AOB. The β-Proteobacterial amoA community composition correlated significantly with water soluble salts in the sediments, whereas the archaeal amoA community composition was correlated more with nitrate concentrations. Quantitative PCR (qPCR) results indicated that the abundance of β-AOB amoA gene (9.11?×?10⁴–6.47?×?10⁵?copies?g^?1 sediment) was always greater than that of AOA amoA gene (7.98?×?10³–3.51?×?10⁵?copies?g^?1 sediment) in all the samples analyzed in this study. The β-Proteobacterial amoA gene abundance was closely related to organic carbon, while no significant correlations were observed between archaeal amoA gene abundance and the environmental factors. Potential nitrification rates were significantly greater in summer than in winter and correlated strongly with the abundance of amoA genes. Additionally, a greater contribution of single amoA gene to potential nitrification occurred in summer (1.03–5.39 pmol?N?copy^?1?day^?1) compared with winter (0.16–0.38 pmol?N?copy^?1?day^?1), suggesting a higher activity of ammonia-oxidizing prokaryotes in warm seasons.