Prostacyclin synthesis stimulating plasma factor in patients with different stages of hepatic coma

In patients with acute hepatic coma the prostacyclin synthesis stimulating plasma factor is significantly enhanced. This increased activity has been verified by using various test systems. However, there is no correlation between the actual plasma factor activity and the coma stage, respectively. The increase of plasma factor activity in these patients might account at least in part for the increased bleeding tendency seen in these patients.     

Albumins stabilize prostaglandin I2

We monitored the chemical stability of prostaglandin I₂ in the presence of different plasma proteins. PGI₂ hydrolyzes by first order reaction kinetics, dependent on the albumin concentration in aqueous buffers. Chemical analysis shows that vertebrate albumins prolong the half-life of prostaglandin I₂. The degree of stabilization varies quantitatively among different species. The results indicate that prostaglandin I₂ may persist in its active form longer than previously supposed in some experimental conditions.     

Comparison of the effects of prostaglandin I2 and prostaglandin E2 stimulation of the rat kidney adenylate cyclase-cyclic AMP systems

Prostacyclin (Prostaglandin I₂) effects on the rat kidney adenylate cyclase-cyclic AMP system were examined. Prostaglandin I₂ and prostaglandin E₂, from 8 · 10^?4 to 8 · ^?7 M stimulated adenylate cyclase to a similar extent in cortex and outer medulla. In inner medulla, prostaglandin I₂ was more effective than prostaglandin E₂ at all concentrations tested. Both prostaglandin I₂ and prostaglandin E₂ were additive with antidiuretic hormone in outer and inner medulla. Prostaglandin I₂ and prostaglandin E₂ were not additive in any area of the kidney, indicating both were working by similar mechanisms. Prostaglandin I₂ stimulation of adenylate cyclase correlated with its ability to increase renal slice cyclic AMP content. Prostaglandin I₂ and prostaglandin E₂ (1.5 · 10^?4 M) elevated cyclic AMP content in cortex and outer medulla slices. In inner medulla, with Santoquin® (0.1 mM) present to suppress endogenous prostaglandin synthesis, prostaglandin I₂ and prostaglandin E₂ increased cyclic AMP content. 6-Ketoprostaglandin F_1α, the stable metabolite of prostaglandin I₂, did not increase adenylate cyclase activity or tissue cyclic AMP content. Thus, prostaglandin I₂ activates renal adenylate cyclase. This suggests that the physiological actions of prostaglandin I₂ may be mediated through the adenylate cyclase-cyclic AMP system.     

Uterine vein prostaglandin levels in late pregnant dogs

We studied the uterine venous plasma concentrations of prostaglandins E₂, F_2α, 15 keto 13,14 dihydro E₂ and 15 keto 13,14 dihydro F_2α in late pregnant dogs in order to evaluate the rates of production and metabolism of prostaglandin E₂ and F_2α in pregnancy $\text{in vivo}$. We used a very specific and sensitive gas chromatography-mass spectrometry assay to measure these prostaglandins. The uterine venous concentrations of prostaglandin E₂ and 15 keto 13,14 dihydro E₂ were 1.35±.27 ng/ml and 1.89±.37 ng/ml, respectively; however, we could not find any prostaglandin F_2α and very little of its plasma metabolite in uterine venous plasma. Since uterine microsomes can generate prostaglandin F_2α and E₂ from endoperoxides, prostaglandin F_2α production $\text{in vivo}$ must be regulated through an enzymatic step after endoperoxide formation. Prostaglandin E₂ is produced by pregnant canine uterus in quantities high enough to have a biological effect in late pregnancy; however, prostaglandin F_2α does not appear to play a role at this stage of pregnancy.     

NADP-linked 15-hydroxyprostaglandin dehydrogenase for prostaglandin D2 in human blood platelets

Two forms of NADP-linked 15-hydroxyprostaglandin dehydrogenase for prostaglandin D₂ were found in the cytosol fraction of human blood platelets. These enzymes were purified by ammonium sulfate fractionation, Blue Sepharose, and Sephadex G-100 column chromatography. The two enzymes differed in molecular weights (65,000 for peak I enzyme and 31,000 for peak II as estimated by gel filtration) and their substrate specificities. The relative rates for reaction with peak I enzyme were: prostaglandin D₂, 100(%); E₂, 14; F_2α, 2; I₂, 29; and B₂, 0; whereas for peak II enzyme, D₂, 100; E₂, 23; F_2α, 61; I₂, 29; and B₂, 131. Prostaglandin D₂ was converted to 15-ketoprostaglandin D₂ and then 13,14-dihydro-15-ketoprostaglandin D₂, which were identified by spectrophotometry and gas chromatography/mass spectrometry, respectively. These metabolites were three orders of magnitude less potent in inhibiting human platelet aggregation than prostaglandin D₂. The results indicated that NADP-linked dehydrogenases participated in the metabolic inactivation of prostaglandin D₂ in the platelets. Furthermore, the dehydrogenase activity for prostaglandin D₂ was high in monkey (0.128 nmol/min · mg at 24 °C) and human platelets (0.066), but was not detectable (less than 0.007) in the rabbit, rat, and chicken. Because prostaglandin D₂, which was demonstrated by several authors to be synthesized in platelet-rich plasma during platelet aggregation, exhibited significant antiaggregatory activity only in human and monkey platelets, these prostaglandin dehydrogenases appear to play a physiological role in the circulatory system.     

Role of prostaglandis in the control of renin secretion in the dog (II)

Infusion of prostaglandin E₁ (PGE₁) into the renal artery of anesthetized dogs (1.03 μg/min) caused increases in urine flow rate (V), renal plasma flow (RPF) and renin secretion rate without any change in mean arterial blood pressure (MABP), whereas infusion of prostaglandin F₂α (PGF_2α), (1.03 μg/min) caused no consistent change in V, RPF, or renin secretion rate. Infusion of prostaglandin E₂ (PGE₂) (1.03 μg/min) into the renal artery of “non-filtering” kidneys caused renin secretion rate to rise from 567.7 ± 152.0 U/min(M ± SEM) during control periods to 1373.6 ± 358.5 U/min after 60 minutes of infusion of PGE₂ (P < 0.01), without significant change in MABP (P > 0.1). The data suggest that PGE₁ and PGE₂ play a role in the control of renin secretion. The data further suggest that PGE may control renin secretion through a direct effect on renin-secreting granular cells.     

Middle ear cell line that maintains vectorial electrolyte transport

The middle ear epithelium plays a major role in keeping the temporal bone cavities fluid-free and air-filled, which is a mandatory condition to allow optimum transmission of the sound vibrations from the tympanic membrane to the inner ear. Previous works have recently established the absorptive function of the middle ear epithelium, using primary cultures derived from Mongolian gerbil (Meriones unguiculatus). Because of the paucity of cells as obtained by enzymatic digestion, we developed a middle ear cell line (MESV) using wild-type SV40 infection of primary culture of Mongolian gerbil's middle ear epithelial cells. Transformation was attested by nuclear expression of SV40 large T antigen, prolonged in vitro passages (presently beyond 50 passages), and tumor-inducing ability when subcutaneously injected in athymic mice. Transport properties were evaluated after the fifteenth passage. MESV cells retained most cardinal properties of the original middle ear epithelial cells: cell polarization was evidenced by the presence of mature junctional complexes that separate the cell membrane in two distinct domains, with apical microvilli at the luminal side, and by vectorial sodium transport responsible for the transepithelial lumen-negative potential difference (?9.3 ± 0.14 mV in culture conditions (n=9), ?2.1 ± 0.25 mV after overnight growth factors and serum deprivation). Short-circuit current was, like in primary cultures, mainly related to a sodium transport occuring through amiloride-sensitive apical sodium channels, since apical addition of amiloride (10^?5 M) reduced I_sc from 7.0 = 1.4 to 0.6 ± 0.1 μA/cm² (P < 0.01, n = 6). Cellular cAMP content was increased by isoproterenol and prostaglandin E2 from 40.5 ± 5.6 to 258.5 ± 17.3 and 55.6 ± 6.2 pmol/mg protein per 5 min, respectively (P < 0.05, n = 10). Isoproterenol and prostaglandin E2 increased I_sc with very similar maximal effects: isoproterenol (10^?4 M) increased I_sc from 5.73 ± 0.31 to 12.77 ± 0.39 μA/cm², while prostaglandin E2 increased I_sc from 5.47 ± 0.21 to 12.87 ± 0.42 (n = 3). Since amiloride (10^?5 M) abolished this stimulation, this may be related to an increase of the electrogenic sodium transepithelial transport. The MESV cell line could provide an interesting tool as a model of middle ear epithelial cells for the study of pathophysiological modulations of ion transport. © 1993 Wiley-Liss, Inc.     

Quantitative analysis of two dinor urinary metabolites of prostaglandin I2

The synthesis of deuterium- and tritium-labeled analogs of 2,3-dinor-6-keto-prostaglandin F_1α and of 6,15-diketo-13,14-dihydro-2,3-dinor-prostaglandin F_1α is described. These analogs were used as internal standards in the assay of the corresponding unlabeled metabolites in human urine by stable isotope dilution and combined gas chromatography-mass spectrometry. In male subjects the 24-h urinary excretion of the two metabolites was found to be 719 ± 264 and 314 ± 115 ng, respectively. The method offers a noninvasive approach to the study of prostaglandin I₂ synthesis in man.     

Effects of combined exenatide and pioglitazone therapy on hepatic fat content in type 2 diabetes

We examined the effects of combined pioglitazone (peroxisome proliferator‐activated receptor‐γ (PPAR‐γ) agonist) and exenatide (GLP‐1 receptor agonist) therapy on hepatic fat content and plasma adiponectin levels in patients with type 2 diabetes (T2DM). Twenty‐one T2DM patients (age = 52 ± 3 years, BMI = 32.0 ± 1.5, hemoglobin A_1c (HbA_1c) = 8.2 ± 0.4%) on diet and/or metformin received additional treatment with either pioglitazone 45 mg/day for 12 months (n = 10) or combined therapy with pioglitazone (45 mg/day) and exenatide (10 µg subcutaneously twice daily) for 12 months (n = 11). At baseline, hepatic fat content and plasma adiponectin levels were similar between the two treatment groups. Pioglitazone reduced fasting plasma glucose (FPG) (P < 0.05), fasting free fatty acid (FFA) (P < 0.05), and HbA_1c (Δ = 1.0%, P < 0.01), while increasing plasma adiponectin concentration by 86% (P < 0.05). Hepatic fat (magnetic resonance spectroscopy (MRS)) was significantly reduced following pioglitazone treatment (11.0 ± 3.1 to 6.5 ± 1.9%, P < 0.05). Plasma triglyceride concentration decreased by 14% (P < 0.05) and body weight increased significantly (Δ = 3.7 kg). Combined pioglitazone and exenatide therapy was associated with a significantly greater increase in plasma adiponectin (Δ = 193%) and a significantly greater decrease in hepatic fat (12.1 ± 1.7 to 4.7 ± 1.3%) and plasma triglyceride (38%) vs. pioglitazone therapy despite the lack of a significant change in body weight (Δ = 0.2 kg). Hepatic injury biomarkers aspartate aminotransferase and alanine aminotransferase (ALT) were significantly decreased by both treatments; however, the reduction in ALT was significantly greater following combined pioglitazone and exenatide therapy. We conclude that combined in patients with T2DM, pioglitazone and exenatide therapy is associated with a greater reduction in hepatic fat content as compared to the addition of pioglitazone therapy (Δ = 61% vs. 41%, P < 0.05).     

The efficacy and acceptability of the ″prostaglandin impact″ in inducing complete abortion during the second week after the missed menstrual period

Twenty-two pregnant patients were exposed, 12±1 days after their missed menstrual period, to a single intrauterine dose of 5mg PG F2_α, to provoke a PG ″Impact″ (PGI) and through it legal abortion. The PGI, delivered through the cervix during 10 minutes, induced 83±9mm Hg contracture in 20±3 minutes. Cyclic IUP reached 102±10mm Hg only in 116±14 minutes, it was then sustained for about 2 hours and subsequently declined.During the evolution of IUP uterine bleeding appeared, progesterone (P) and estradiol 17_β (E₂) started to decrease and continued decreasing. At 24 hours after PGI, P-withdrawal was 44% (P<0.05), bleeding continued and cervical dilatation approximated 1cm. Subsequently uterine bleeding (containing tissue fragments) continued and out of 22 women 20 aborted completely. After 3–5 days bleeding declined, the pregnancy tests became negative and normal menstrual periods one month after PGI provided the desired end points of the study. Complete abortions, simulating the symptoms of delayed menstrual periods, had a high Abortion Score of 95 and in the sedated patients the side effects were infrequent, mild and acceptable. This clinical outcome encourages extensive field trials.None of the patients aborted during the short lived period of exogenous PG stimulation. However, the continued steroid withdrawals indicated that PGI damaged the endocrine function of the ovum and the luteotrophic support of the corpus luteum. Therefore, if it was PG which completed abortion eventually, it had to be the endogenous compound, having become effective in physiological concentrations, due to the threshold-lowering action of P-withdrawal.     

Prostaglandin F2α concentration in genital tract secretions of dairy bulls

Prostaglandin F_2α (PGF_2α) concentrations in genital tract secretions of conscious dairy bulls were determined by radioimmunoassay procedures and compared with peripheral blood plasma levels. The mean (± SD) PGF_2α concentration of coccygeal venous blood plasma from four bulls was 0.14 ± 0.05 ng/ml. Values for rete testis fluid and seminal plasma were the same, namely 0.17 ± 0.01 ng/ml (n = 5) and 0.17 ± 0.02 ng/ml (n = 4), respectively. However, the PGF_2α level in cauda epididymal plasma was 1.61 ± 0.41 ng/ml, or over 8 to 10 times (P < 0.01) the concentration of any other fluid studied.Added PGF_2α had no effect on the endogenous oxygen consumption of washed cauda epididymal spermatozoa or on the oxidative and glycolytic activities of washed ejaculated spermatozoa in vitro. No evidence was obtained suggesting that the prostaglandin may interact with the stimulatory effect of added testosterone or phosphatidylinositol (PI) on the motility, respiration or glucose uptake of ejaculated spermatozoa.     

苦参碱对棉铃虫幼虫神经细胞钠通道的影响

用全细胞膜片钳技术研究了生物碱类植物杀虫剂苦参碱对棉铃虫Helicoverpa armigera幼虫离体培养中枢神经细胞钠离子通道门控过程的影响。结果表明： 苦参碱对棉铃虫幼虫神经细胞所表达的TTX （tetradotoxin, 河豚毒素)敏感钠通道具有浓度依赖性阻滞作用,1,10和100 μmol/L的苦参碱作用5 min后,分别使钠电流峰值较给药前下降（12.49±1.67）%、（18.79±2.16）%和（43.15±8.17）% (n=8, P<0.05)。苦参碱使钠电流的电流 电压关系曲线上移,但并不改变其激活电压、峰电压和电流电压关系曲线的形状。苦参碱对钠通道的阻滞作用可能是其具有某些毒理效应的离子基础。     

Platelet activating factor-induced aggregation of calf platelets: Apparent positive cooperativity in the kinetics and non competitive inhibition by diltiazem

Aggregation of calf platelets by platelet activating factor was characterized by a spectrophotometric method. The aggregation kinetics of both platelet-rich plasma and purified platelets showed concave up double-reciprocal plots and linear Hill plots withh > 1 (1.7 ± 02) consistent with positive cooperativity. Comparable values of maximum rates of aggregation(R) were obtained with platelet-rich plasma (0.25 ± 0.08) and purified platelets (0.28 ± 0.18) but the half-maximal saturation concentration (S_0.5) differed greatly between platelet-rich plasma (6 ± 3 nM) and purified platelets (0.28 ± 0.18 nM). An Arrhenius activation energy of 21 ±2 kcal/mol was found for aggregation of purified platelets. Diltiazem was inhibitory with half-maximal inhibitory concentration (I_0.5) of 4 M but the inhibition was not competitive. Diltiazem inhibited rates but not the extent of shape-change. The receptor-antagonist and sulphydryl reagent N-ethylmaleimide and the platelet antagonistic omega-3-fatty acid, 5,8,11,14,17-eicosa pentaenoic acid, inhibited both rates and extent of shape-change reactions and inhibited aggregation competitively (I_0.5 ∼ 5 M). Eicosa pentaenoic acid at > 25 M could abolish shape-change reactions and at 50 M served as an activator of platelets and the activation was enhanced by aspirin (1 mM). Although N-ethylmaleimide at > 20 M could also induce platelet activation it failed to induce aggregation and aspirin had no effect on the shape-change reactions induced by it.     

Novel sulphonamides incorporating triazene moieties show powerful carbonic anhydrase I and II inhibitory properties

Abstract

A series of compounds incorporating 3-(3-(2/3/4-substituted phenyl)triaz-1-en-1-yl) benzenesulfonamide moieties were synthesised and their chemical structure was confirmed by physico-chemical methods. Carbonic anhydrase (CA, EC 4.2.1.1) inhibitory effects of the compounds were evaluated against human isoforms hCA I and II. K_I values of these sulphonamides were in the range of 21?±?4–72?±?2?nM towards hCA I and in the range of 16?±?6–40?±?2?nM against hCA II. The 4-fluoro substituted derivative might be considered as an interesting lead due to its effective inhibitory action against both hCA I and hCA II (K_Is of 21?nM), a profile rarely seen among other sulphonamide CA inhibitors, making it of interest in systems where the activity of the two cytosolic isoforms is dysregulated.     

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

Thromboxane A₂ plays and 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 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 cycyooxygenase enzyme. Parallel experiments with washed platelet preparations suggest that they may be an inadequate mode for studying relationship between the platelet cyclooxygenase and platelet function.     

Urinary prostaglandin e2 excretion in chronic renal disease

To ascertain whether renal prostaglandin E₂ production is impaired in chronic renal disease we measured urinary prostaglandin E₂ in 25 female renal patients, and in 38 female normal volunteers. Prostaglandin E₂ averaged 337 ± 63 ng/24 hr in 25 female renal patients, which was significantly higher (P < 0.01) than values in normal female subjects (166 ± 17 ng/24 hr). None of the renal patients excreted less than 50 ng/24 hr, whereas normals when treated with indomethacin averaged 30 ± 10 ng/24 hr (n=8). There were no significant correlations between prostaglandin E₂ and creatinine clearance, or urine volume. No difference in prostaglandin E₂ was encountered between hypertensive and normotensive renal patients. Our results indicate that renal prostaglandin E₂ production is increased rather than decreased in renal disease. The failure to demonstrate decreased urinary prostaglandin E2 excretion in renal hypertension sharply contrasts with the situation in essential hypertension.     

Dietary Iodine and Selenium Affected the mRNA Expression Levels of Skin Monodeiodinase (II, III) in Liaoning Cashmere Goats

Livestock are frequently provided nutrient-depleted diets, which can negatively impact animal health and productivity. In our previous trial, we found that iodine (I) supplementation (not selenium (Se)) could increase cashmere production. In order to explore the role of I and Se in cashmere growth, we investigated the effects of dietary I and Se supplementation in Liaoning cashmere goats. Serum thyroid hormone status and the mRNA expression levels of skin monodeiodinase (MDII, MDIII) were measured during the cashmere fiber growth period. Forty-eight 2.5-year-old Liaoning cashmere goats (38.6?±?2.65 kg BW) were divided into six equal groups, and their diets were supplemented with I (0, 2, or 4 mg/kg DM) and Se (0 or 1 mg/kg DM) in a 2?×?3 factorial treatment design. The six treatment groups were: I₀Se₀, I₂Se₀, I₄Se₀, I₀Se₁, I₂Se₁, and I₄Se₁. Concentrations of I and Se in the basal diet (group I₀Se₀) were 0.67 and 0.09 mg/kg DM, respectively. The trial started in September of 2009 and lasted 70 days. For every measured parameter, supplemental Se had no significant effect on thyroid hormones, but improved the mRNA expression levels of skin MDIII (P?<?0.01). However, supplemental I increased levels of thyroid hormones (thyroxine and triiodothyronine) and improved the mRNA expression levels of skin MDII (P?<?0.05). These results show that the addition of I to cashmere goat feedstock may be an effective means of increasing cashmere production through thyroid hormones regulating the mRNA expression of skin MDII.     

The antioxidant effect of ubiquinone and combined therapy on mitochondrial function in blood cells in non-proliferative diabetic retinopathy: A randomized,double-blind,phase IIa,placebo-controlled study

Objectives: To evaluate the effect of ubiquinone and combined antioxidant therapy on mitochondrial function in non-proliferative diabetic retinopathy (NPDR) in a randomized, double-blind, phase IIa, placebo-controlled, clinical trial. Three groups of 20 patients were formed: Group 1, ubiquinone; Group 2, combined therapy; and Group 3, placebo (one daily dose for 6 months).

Methods: Fluidity of the submitochondrial membrane in platelets was determined by examining intensity of fluorescence between the monomer (I_m) and excimer (I_e). Hydrolytic activity of the mitochondrial F₀F₁-ATPase was evaluated with the spectrophotometric method.

Results: Normal, baseline submitochondrial membrane fluidity, 0.24 ± 0.01 I_e/I_m, was significantly diminished in the three study groups vs. normal values (P < 0.0001); placebo, 0.14 ± 0.01 I_e/I_m; ubiquinone, 0.14 ± 0.01 I_e/I_m; and combined therapy, 0.13 ± 0.00 I_e/I_m. Afterward, it increased significantly (P < 0.0001), the ubiquinone group 0.22 ± 0.01 I_e/I_m, combined therapy group, 0.19 ± 0.01 I_e/I_m; with no changes the placebo group. Baseline hydrolytic activity of the F₀F₁-ATPase enzyme increased in the three study groups vs. normal values (184.50 ± 7.84 nmol PO₄), placebo, 304.12 ± 22.83 nmol PO₄ (P < 0.002); ubiquinone, 312.41 ± 25.63 nmol PO₄ (P < 0.009); and combined therapy, 371.28 ± 33.50 nmol PO₄ (P < 0.002). Afterward, a significant decrease the enzymatic activity: ubiquinone, 213.25 ± 14.19 nmol PO₄ (P < 0.001); and combined therapy, 225.55 ± 14.48 nmol PO₄ (P < 0.0001).

Discussion: Mitochondrial dysfunction significantly improved in groups of NPDR patients treated with antioxidants.     

A comparison of the pulmonary, renal and hepatic extractions of PGI2 and PGE2 - PGI2 a potential circulating hormone.

Prostaglandin (PG) I₂ and PGE₂ were infused into the aortic arch, femoral vein, renal artery and portal vein in anesthetized dogs over a dose range to produce a steady decrease in systemic blood pressure after 10 mins infusion. Parallel log dose-response relationships were observed with both PGI₂ and PGE₂. PGE₂ was a more potent depressor than PGI₂ when infused into the aortic arch. The doses to reduce blood pressure by 5 mm Hg were used to calculate the extraction of the compounds by the lungs, kidney and liver. The pulmonary extraction of PGE₂ was 96 ± 2% and was essentially complete following combined pulmonary and renal or pulmonary and hepatic extraction. In contrast, there was no significant pulmonary extraction of PGI₂. Combined renal and pulmonary extraction was 43 ± 11% and combined hepatic and pulmonary extraction 87 ± 5%. These results indicate a marked difference in the organ metabolising capacity for PGE₂ and PGI₂. Since PGI₂ has been shown to be produced both in the kidney and stomach it is possible that PGI₂ produced endogenously could pass into the circulation and exert systemic pharmacological effects.