AEV-transformed erythroleukemia cell induced differentiation: Expression of specific cell membrane antigenic molecules

Summary A simultaneous decay of the expression of Im 140 kDa, Im 150 kDa and Im 160 kDa high MW membrane antigens, concomitant with the cell proliferation arrest, was observed during erythropoietin induced differentiation ofts 34 AEV-transformed erythroid cells cultivated at the restrictive temperature. Expression of embryo-immature antigens was maintained during induced differentiation of erythroleukemia cells, but their MW shifted from 50 to 48 kDa, which corresponds to the MW of embryo-immature antigens detected on normal erythroid cells. In the absence of erythropoietin at the restrictive temperature, conditions under which thets 34 AEV-transformed erythroid cells fail to differentiate and maintain their capacity to proliferate, the expression of high MW antigens as well as the expression of embryoimmature antigens remained unaffected. Therefore, it is shown that the expression of specific membrane antigens is modulated under conditions rendering the erythroleukemia cell differentiation process possible.With 3 Figures     

A 2-year follow-up of quality of life, pain, and psychosocial factors in patients with chronic prostatitis/chronic pelvic pain syndrome and their spouses

Objectives

There are two objectives: (1) Examine quality of life (QoL) and mood between chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) patients and spouses over a 2-year period; (2) Longitudinally assess CP/CPPS patient pain, disability, and pain catastrophizing over a 2-year period.

Methods

Forty-four CP/CPPS diagnosed men and their spouses participated. Patients completed demographics, QoL, depression, anxiety, pain, disability, and catastrophizing across the study. Spouses completed QoL, depression, and anxiety. Patients/spouses were not different in education, but patients were older (49 years; SD = 9.56). The average symptom duration was 8.68 (SD = 7.61). Couples were married or common law, and majority of patients were employed. Due to attrition, approximately 21 couples provided analyzable data.

Results

Patients and spouses physical QoL did not statistically differ over time from one another, and both increased over the study period. Mental QoL increased over time, but patients reported lower QoL. Patients reported more depression and anxiety, but both measures remained stable over time for spouses and patients. Finally, patient only analyses showed that disability did decrease over time from a high at 6 months, but pain and catastrophizing showed stability over the 2 years.

Conclusions

Patients reported worse mental QoL, depression, and anxiety compared to spouses, and spouses reported significant stable levels of depression and anxiety similar to patients. Further, patient catastrophizing, pain, and disability did not reduce over the 2-year assessment period. These results provide further impetus for the development and implementation of mental health strategies alongside continued medical efforts in couples suffering from CP/CPPS.     

Ammodytase, a metalloprotease from Vipera ammodytes ammodytes venom, possesses strong fibrinolytic activity

Ammodytase, a high molecular mass metalloproteinase with fibrinogenolytic and fibrinolytic activities, was purified from long-nosed viper (Vipera ammodytes ammodytes) venom by gel filtration, affinity and ion-exchange chromatographies. The enzyme is a single-chain glycoprotein with apparent molecular mass of 70 kDa and isoelectric point of 6.6. Ammodytase shows very weak hemorrhagic activity, and only at doses higher than 20 μg. Consistent with this, it partially degrades some components of the extracellular matrix in vitro. It cleaves the Aα-chain of fibrinogen preferentially at peptide bonds Glu⁴⁴¹-Leu⁴⁴² and Glu⁵³⁹-Phe⁵⁴⁰. Its preference for bulky and hydrophobic amino acids at the P1′ position in substrates is demonstrated by its hydrolysis of only the Gln⁴-His⁵ and Tyr¹⁶-Leu¹⁷ bonds in the B-chain of insulin. Ammodytase is able to dissolve fibrin clots. It neither activates nor degrades plasminogen and prothrombin, and has no effect on collagen- or ADP-induced platelet aggregation in vitro. LC/MS and MS/MS analyses of its tryptic fragments demonstrated that ammodytase is a P-III class snake venom metalloproteinase composed of metalloproteinase, disintegrin-like and cysteine-rich domains. Its similarity to hemorrhagins from V. a. ammodytes venom, accompanied by very low toxicity, makes ammodytase a promising candidate as an antigen to prepare antisera against these most dangerous components of the viper's venom. Moreover, its ability to degrade fibrin clots suggests its clinical use as an antithrombotic agent.     

Educating Adolescents About Healthy Sleep: Experimental Study of Effectiveness of Educational Leaflet

Aim

To evaluate how exposure to educational leaflet about healthy sleep affects knowledge about sleep in adolescents.

Methods

The study included students aged 15-18 years from 12 high schools (1209 participants; 85% of eligible study population). Multistage sampling was used and the selected schools were randomly assigned into two intervention groups and two control groups, according to the Solomon experimental design. Intervention groups received educational leaflets and control groups did not. In one of the intervention groups and one of the control groups, pre-testing of knowledge about sleep was performed. Students answered the Sleep Knowledge Test, which was constructed in accordance with the information on the leaflet. Data were analyzed by four-way ANOVA and additional analyses of simple main effects were performed.

Results

Positive effect of educational leaflet was found in students aged 15 (F = 28.46; P < 0.001), 16 (F = 5.74; P = 0.017), and 17 (F = 17.17; P < 0.001), but there was no effect in students aged 18 (P = 0.467). In male students, positive effect of the leaflet was found only in the group that had not been pre-tested (F = 6.29; P = 0.012), while in female students, it was found in both pre-tested (F = 26.24; P < 0.001) and not pre-tested group (F = 17.36; P < 0.001), with greater effect in pre-tested group (F = 5.70; P = 0.017). Female students generally showed better knowledge about sleep than male students (F = 95.95; P < 0.001).

Conclusion

Educational leaflets can be an effective first step in educating younger high school students about healthy sleep, with the method being more effective in female adolescents.Sleep education has been used as a method of primary and secondary prevention of sleep problems in all age groups (1-3). An especially vulnerable age group are adolescents who frequently have poor sleep habits and suffer from sleep deprivation (4-6). In adolescents, insufficient sleep, inadequate sleep quality, and irregular sleep patterns are associated with daytime sleepiness, negative moods, increased likelihood of stimulant use, higher levels of risk-taking behavior, poor school performance, and increased risk of unintentional injuries (7-10). As an US study has shown, sleepiness was the major causal factor in many traffic accidents and more than 50% of sleep-related crashes involved drivers aged 25 or younger (11).Having in mind that adolescence is not only a period when sleep problems arise, but also a period when many life habits are established, adolescent education about healthy sleep becomes an important task. Different educational programs and public educational campaigns have been organized to increase the knowledge about healthy sleep and consequences of sleepiness in adolescents and their parents and teachers (12-14). The effects of such educational programs on adolescents’ sleep knowledge and characteristics have been described by several studies (2,12).Another way to increase knowledge about sleep and to foster positive behavioral changes regarding sleep in adolescents are public education campaigns. In order to achieve these goals, effective educational methods need to be developed and a systematic evaluation of their effectiveness performed. In this study, we evaluated the effect of our educational effort to increase adolescents’ knowledge about sleep. The method we used was exposure to leaflets, which is a commonly used method in public health campaigns. Since some studies have shown sex differences in school performance (15,16), we expected that sleep education would have a different effect on knowledge about sleep in boys than in girls. The effect of age on sleep education may also be expected because of possible differences in the basic knowledge about sleep in students of different age.     

Purification and characterisation of two hemorrhagic metalloproteinases from the venom of the long-nosed viper, Vipera ammodytes ammodytes.

Two hemorrhagic proteins, VaH1 and VaH2, have been purified from Vipera ammodytes ammodytes venom. They are monomeric glycoproteins of an apparent molecular mass of 70kDa and multiple isoelectric points around pH 5.5. Both molecules are proteolytically active against azocasein as substrate. VaH1, which was characterised in detail, showed maximum activity at pH 7.5. Ethylenediaminetetraacetic acid eliminated the proteolytic as well as the hemorrhagic activity of VaH1 while iodoacetamide, phenylmethylsulfonyl fluoride and pepstatin A, inhibitors of cysteine, serine and aspartic proteinases respectively, had no effect. VaH1 is therefore a metalloproteinase whose hemorrhagic activity is very likely the result of its proteolytic activity. VaH1 is a fibrinogenase, hydrolysing exclusively the Aalpha-chain of fibrinogen. In the B-chain of insulin it cleaved with a high preference the bond between Ala(14) and Leu(15). Based on its molecular mass, VaH1 (as well as VaH2) is a Class P-III metalloproteinase. Partial amino acid sequences of its CNBr fragments demonstrated a high level of identity with the reprolysin subfamily of zinc-metalloproteinases.     

中、低收入国家卫生智囊团的实践经验影响政策改变

近年来,中、低收入国家独立卫生政策研究机构的数量与日俱增,原因在于政府研究能力有限和民主化进程中的压力。本研究的目标是：（1）调查中﹑低收入国家中卫生政策研究机构对卫生政策的议程设置﹑制定﹑执行、监管和评估所作的贡献;（2）评估包括组织形式和结构在内的哪些因素支持中﹑低收入国家的卫生政策研究机构对卫生政策发挥积极作用。本研究在孟加拉﹑加纳﹑印度﹑南非﹑乌干达和越南选取了6家卫生政策研究机构开展案例研究,研究对象包括两个非政府组织、两所大学和两个政府办政策研究机构。案例研究通过文献查阅、财务信息分析、对工作人员和其他利益相关人员进行半结构式访谈,以及对结论草案进行多次反馈等方式开展。其中有些机构对他们各自国家的政策发展作出了巨大贡献。这些机构都积极建言献策,多数从事与政策相关的研究,而开展政治对话﹑系统评价或委托性研究的机构则相对较少。这些机构所开展的工作大多以政府或出资人的需求为导向,多数机构的主要成果一般为研究报告,经常与面向政府官员的口头汇报相结合。在支持对政策的有效参与方面,有几个关键因素,其中包括支持性的政策环境﹑管理和财务的相对独立性,以及与决策者建立可增进信任和影响的密切关系。当研究机构与政府之间的正式关系未处在重要位置时,政府内部单位则面临相当大的困难。     

An agonist-induced switch in G protein coupling of the gonadotropin-releasing hormone receptor regulates pulsatile neuropeptide secretion

The pulsatile secretion of gonadotropin-releasing hormone (GnRH) from normal and immortalized hypothalamic GnRH neurons is highly calcium-dependent and is stimulated by cAMP. It is also influenced by agonist activation of the endogenous GnRH receptor (GnRH-R), which couples to G(q/11) as indicated by release of membrane-bound alpha(q/11) subunits and increased inositol phosphate/Ca(2+) signaling. Conversely, GnRH antagonists increase membrane-associated alpha(q/11) subunits and abolish pulsatile GnRH secretion. GnRH also stimulates cAMP production but at high concentrations has a pertussis toxin-sensitive inhibitory effect, indicative of receptor coupling to G(i). Coupling of the agonist-activated GnRH-R to both G(s) and G(i) proteins was demonstrated by the ability of nanomolar GnRH concentrations to reduce membrane-associated alpha(s) and alpha(i3) levels and of higher concentrations to diminish alpha(i3) levels. Conversely, alpha(i3) was increased during GnRH antagonist and pertussis toxin treatment, with concomitant loss of pulsatile GnRH secretion. In cholera toxin-treated GnRH neurons, decreases in alpha(s) immunoreactivity and increases in cAMP production paralleled the responses to nanomolar GnRH concentrations. Treatment with cholera toxin and 8-bromo-cAMP amplified episodic GnRH pulses but did not affect their frequency. These findings suggest that an agonist concentration-dependent switch in coupling of the GnRH-R between specific G proteins modulates neuronal Ca(2+) signaling via G(s)-cAMP stimulatory and G(i)-cAMP inhibitory mechanisms. Activation of G(i) may also inhibit GnRH neuronal function and episodic secretion by regulating membrane ion currents. This autocrine mechanism could serve as a timer to determine the frequency of pulsatile GnRH release by regulating Ca(2+)- and cAMP-dependent signaling and GnRH neuronal firing.     

Safety assessment of N-acetyl-l-threonine

N-acetyl-l-threonine (NAT) is a dietary constituent that has been identified at low concentrations (<1 μg/g fresh weight) in numerous foods. The current paper reports the outcome of toxicology studies conducted to assess the effects of NAT. No evidence of mutagenicity or genotoxicity was observed in in vitro bacterial or in vivo mammalian studies. No mortalities or evidence of adverse effects were observed in Sprague–Dawley (SD) rats following acute oral administration of 2000 mg of NAT/kg of body weight (kg of bw). A 28-day repeated dose toxicity study was conducted in SD rats by incorporating NAT into diets at concentrations targeting up to 1000 mg of NAT/kg of bw/day. All rats survived until scheduled sacrifice and no biologically significant differences were observed in any of the NAT treatment groups for body weights, feed consumption, clinical signs, behavioral, ophthalmology, hematology, coagulation, clinical chemistry, organ weights, or gross or microscopic changes. Based on these results, NAT does not represent a risk for mutagenicity or genotoxicity, is not acutely toxic, and the no-observed-adverse-effect-level (NOAEL) for systemic toxicity from repeated dose dietary exposure to NAT is 848.5 and 913.6 mg/kg of bw/day for male and female SD rats, respectively.