Neuroendocrine regulation of carbonic anhydrase expression in the gills of the euryhaline green crab, Carcinus maenas

The relationship between branchial carbonic anhydrase (CA) activity, CA gene expression and salinity, and potential mechanisms of regulation, was investigated in the euryhaline green crab, Carcinus maenas, acclimated to 33 ppt and transferred to 10 ppt, and the stenohaline rock crab, Cancer irroratus, acclimated to 32 ppt and transferred to 18 ppt. CA activity in green crabs acclimated to high and low salinity was a function of CA mRNA expression, with low salinity exposure resulting in an increase in both CA expression and activity. Eyestalk ablation (ESA) in green crabs acclimated to high salinity resulted in an increase in CA expression in the posterior, ion-transporting gills, in the absence of the low salinity stimulus. There were no changes in CA activity or expression in the anterior, respiratory gills. ESA also potentiated low salinity-stimulated CA induction, again, only in posterior gills. There were no changes in CA activity in any gills of Cancer irroratus, in response to either ESA or low salinity. These results suggest that CA expression in euryhaline, osmoregulating species, is under inhibitory regulation by a putative repressor found in the eyestalk, and that this mechanism is absent in stenohaline, osmoconforming species. CA expression is maintained at low, baseline levels in crabs acclimated to high salinity by the presence and action of this compound. The effects of the repressor appear to be reduced upon exposure to low salinity, allowing CA induction to occur.     

Macrolide resistance and <Emphasis Type="Italic">In Vitro</Emphasis> selection of resistance to antibiotics in <Emphasis Type="Italic">Lactobacillus</Emphasis> isolates

Spreading of resistance to antibiotics is of great concern due to the increasing rate of isolation of multiresistant pathogens. Since commensal bacteria may transfer determinants of resistance to pathogens, studies on development of resistance should include also lactobacilli. Resistance to macrolides, penicillins and tetracycline was determined in 40 isolates of Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus crispatus, and Lactobacillus casei isolated from faeces of apparently healthy volunteers. Frequency of mutation and changes in susceptibility after serial exposure to these antibiotics at concentrations of 4× and 8× MIC were evaluated in susceptible isolates. Acquired resistance was defined as an increment in MIC values of at least four times in respect to the pre-selection values. Resistance to macrolides and/or tetracycline was identified in 14 and 4 isolates, respectively. ermB gene and A2058G mutation in 23S rRNA were detected in macrolide resistant isolates. Frequencies of mutation of susceptible isolates (n=26) were lower for ampicillin and erythromycin than for tetracycline. Serial exposure to antibiotics led to selection of resistant mutants. However, acquired resistance was rather unstable and was lost after subcultures in antibiotic-free medium in most mutants. Resistance to erythromycin was associated to a A2058G mutation in 23S rRNA. In conclusion, results indicate that resistance to macrolides and tetracycline is present among intestinal lactobacilli. Decrease in susceptibility following serial exposure to antibiotics might occur in lactobacilli, in a strain- and antibiotic-dependent way. Since lactobacilli are often used as probiotics, their ability to acquire resistance should be evaluated for isolates candidate to be included in probiotics based products.     

Effects of estrogen-depletion on rat casein gene expression

Mammary tissue from rats that had been ovariectomized and adrenalectomized 4 weeks previously was compared to that from intact rats in terms of epithelial content and hormone-responsiveness in vitro. The endocrinectomy resulted in about a 30% enlargement of the gland, but led to a loss of only about 12% of the epithelium. This estrogen-depleted epithelium was able to acquire full responsiveness in vitro to insulin in terms of the accumulation of alpha-aminoisobutyric acid, and induction of glucose-6-phosphate and gluconate-6-phosphate dehydrogenases. It was also fully responsive to cortisol in relation to the induction of NADH-cytochrome C reductase, and to prolactin in terms of total RNA synthesis. However, estrogen-depletion led to an 82% loss in the ability of a unit amount of the epithelium to synthesize casein in response to these 3 hormones, and to a similar loss in relation to the accumulation of 25K casein mRNA. Estrogen administration in vivo could prevent and reverse the casein lesion. The disparity between constitutive and casein hormone-responsiveness in the absence of estrogen is discussed in relation to cell commitment.     

水稻品种对美国稻瘟病小种IB-33、IB-45和IE-1的抗性遗传

1995年10月至1997年11月,在美国阿肯色大学水稻研究推广中心,用水稻品种LA110和Jasmine-85与水稻品种Teqing、Katy、Mars、LaGrue和Newbonnet进行不完全双列杂交,对其杂交后代和亲本用美国3个主要稻瘟病菌小种(以下简称小种)IB-33、IB-45和IE-1进行接种鉴定和遗传分析研究.结果表明:亲本LA110、Jasmine-85、Teqing抗所有3个小种.Katy抗小种IB-45和IE-1,感小种IB-33.Mars抗小种IE-1,感小种IB-33和IB-45.LaGrue感所有3个小种.Newbonnet抗小种IB-45,感小种IB-33和IE-1.所有抗病亲本的抗病基因,其F1分别对相应小种呈现显性抗病性.抗病亲本杂交,LA110与Jasmine-85对小种IB-33,LA110与Teqing、Jasmine-85对小种IE-1,及Jasmine-85与Teqing对小种IE-1,是等位的抗病基因.LA110与Teqing对小种IB-33,及Jasmine-85与Teqing对小种IB-33,分别存在三对独立遗传的显性抗病基因.LA110与Teqing、Katy、Newbonnet、Jasmine-85对小种IB-45,Jasmine-85与Teqing、Katy、Newbonnet对小种IB-45,LA110与Katy、Mars对小种IE-1,Jasmine-85与Katy、Mars对小种IE-1,分别存在两对独立遗传的显性抗性基因.抗病亲本LA110或Jasmine-85与感病亲本Mars对小种IB-33,抗病亲本LA110与感病亲本Mars对小种IB-45,具有两对显性互补抗病基因,当两对显性抗病基因同时存在时,表现出抗性.抗病亲本LA110或Jasmine-85与感病亲本Katy、LaGrue、Newbonnet对小种IB-33,抗病亲本LA110与感病亲本LaGrue对小种IB-45,抗病亲本Jasmine-85与感病亲本Mars、LaGrue对小种IB-45,抗病亲本LA110或Jasmine-85与感病亲本LaGrue、Newbonnet对小种IE-1,分别存在一对显性抗病基因.两个亲本正、反交的遗传表现一致.本文也讨论了LA110、Teqing和Jasmine-85三个抗病品种在美国水稻抗病育种中利用的可能性.     

Cell wall and protoplast isoperoxidases in tobacco plants in relation to mechanical injury and infection with tobacco mosaic virus

Leaves and pith of Turkish, Wisconsin 38, and Samsun NN tobacco (Nicotiana tabacum) varieties, which differ in their sensitivity to tobacco mosaic virus, showed the same qualitative isoperoxidase patterns and a similar distribution of distinctive isoperoxidases between the cell protoplast and wall-free, ionically, and covalently bound fractions. No changes in the qualitative isoenzyme spectrum were found in relation to age, mechanical injury, or leaf infection with tobacco mosaic virus. The distinctive isoperoxidases which reacted to infection were the same as those responsive to mechanical injury, confirming that the enzyme reaction to infection results from a nonspecific response to injury. The increase in peroxidase activity in response to infection or mechanical injury, or both, was greater in young tissue than in the older ones. The great increase in Samsun NN leaves and no increase in those of the two other varieties in response to infection may be due to differences in the degree to which the pathogen affected processes controlling the nonspecific peroxidase reaction to injury. Peroxidase development in the infected Samsun NN leaves was due to isoenzymes which form the wall-bound fraction in very young tissues, and to those which increase in activity with aging in the protoplast and wall-free fractions. In mechanically injured tissue, only the first group of isoenzymes increased in activity. In Samsun NN plants, the increased peroxidase activity in upper intact leaves above the infected ones was only due to isoenzymes whose activity increases with both normal and virus-accelerated senescence. Peroxidase reaction to challenge inoculation in these leaves was the same whether the lower ones were intact, infected and/or mechanically injured. Thus, the induced systemic resistance to tobacco mosaic virus may be due to other than peroxidase factors.     

Strategies adopted by the mudskipper Boleophthalmus boddaerti to survive sulfide exposure in normoxia or hypoxia

The effects of sulfide on the energy metabolism of Boleophthalmus boddaerti in normoxia and hypoxia were examined. The 24-, 48-, and 96-h LC50 values of sulfide for B. boddaerti with body weight ranging from 11.6 to 14.2 g were 0.786, 0.567, and 0.467 mM, respectively. The tolerance of B. boddaerti to sulfide was not due to the presence of a sulfide-insensitive cytochrome c oxidase. There was no accumulation of lactate in the muscle and liver of specimens exposed to sulfide in normoxia. In addition, the levels of ATP, AMP, and energy charge in both the muscle and the liver were unaffected. These results indicate that B. boddaerti was able to sustain the energy supply required for its metabolic needs via mainly aerobic respiration when exposed to sulfide (up to 0.4 mM) in normoxia. Exposure of B. boddaerti simultaneously to hypoxia and 0.2 mM sulfide for 48 h resulted in decreases in the ATP levels in the muscle and liver. However, the energy charge in both tissues remained unchanged, and the level of lactate accumulated in the muscle was too low to have any major contribution to the energy budget of the fish. Our results reveal that B. boddaerti possesses inducible mechanisms to detoxify sulfide in an ample supply or a lack of O2. In normoxia, it detoxified sulfide to sulfate, sulfite, and thiosulfate. There were significant increases in the activities of sulfide oxidase in the muscle and liver of specimens exposed to sulfide, with that in the liver being >13-fold higher than that in the muscle. However, in hypoxia, sulfide oxidase activity in the liver was suppressed in response to environmental sulfide. In such conditions, there were significant increases in the activities of sulfane sulfur-forming enzyme(s) in the muscle and liver that were not observed in specimens exposed to sulfide in normoxia. Correspondingly, there were no changes in the levels of sulfate or sulfite in the muscle or liver. Instead, B. boddaerti detoxified sulfide mainly to sulfane sulfur in hypoxia. In conclusion, B. boddaerti was able to activate different mechanisms to detoxify sulfide, producing different types of detoxification products in normoxia and hypoxia.     

湖北省褐飞虱对吡虫啉、噻嗪酮及氟虫腈的抗药性监测

采用稻茎浸渍法监测湖北省武穴、天门、监利、通城、孝感、公安及枣阳七地褐飞虱Nilaparvata lugens(Sta1)田间种群对吡虫啉、噻嗪酮、氟虫腈的抗药性。结果表明:2006、2007和2008年3年7地褐飞虱种群对吡虫啉的抗性分别为98.89～389.19倍、69.00～153.33倍和56.32～116.89倍,达高水平至极高水平抗性,但3年期间总体抗性水平呈现一定程度的下降趋势;2007和2008年2年7地褐飞虱对噻嗪酮的抗性分别为13.39～41.06倍和6.94～20.44倍,达低水平至高水平抗性;2007和2008年2年7地褐飞虱对氟虫腈抗性分别为0.97～2.64倍和0.63～1.42倍,为敏感阶段。此外,还对褐飞虱的治理进行讨论。     

Impact of vitamin E or selenium deficiency on nematode-induced alterations in murine intestinal function

The effects of deficiencies in the antioxidant nutrients, vitamin E and selenium, on the host response to gastrointestinal nematode infection are unknown. The aim of the study was to determine the effect of antioxidant deficiencies on nematode-induced alterations in intestinal function in mice. BALB/c mice were fed control diets or diets deficient in selenium or vitamin E and the response to a secondary challenge inoculation with Heligmosomoides polygyrus was determined. Egg and worm counts were assessed to determine host resistance. Sections of jejunum were mounted in Ussing chambers to measure changes in permeability, absorption, and secretion, or suspended in organ baths to determine smooth muscle contraction. Both selenium and vitamin E deficient diets reduced resistance to helminth infection. Vitamin E, but not selenium, deficiency prevented nematode-induced decreases in glucose absorption and hyper-contractility of smooth muscle. Thus, vitamin E status is an important factor in the physiological response to intestinal nematode infection and may contribute to antioxidant-dependent protective mechanisms in the small intestine.     

稻虾共作水体浮游植物群落结构特征分析

为认识稻虾共作水体浮游植物群落结构特征, 于2016年4月至12月对江汉平原4处稻虾共作水体浮游植物和理化因子开展了逐月调查与分析。共鉴定出浮游植物7门124种, 其中绿藻78种、蓝藻16种、硅藻15种、裸藻3种、隐藻2种、甲藻7种、金藻3种。6—9月浮游植物的种类数、细胞密度、叶绿素a含量达到最大值, 最小值出现在12月份; 浮游植物细胞密度波动范围在1.37×105—2.93×108 cells/L, 叶绿素a含量的变化范围为0.15—208.60 μg/L。调查期间浮游植物的优势种共28种, 主要优势种有颤藻、蓝纤维藻、微囊藻、小球藻、隐藻等。浮游植物Shannon-Wiener多样性指数周年变化范围为0.64—6.3, 多样性指数最高出现在10月份, 最低出现在8月份。结果显示, 稻虾共作水体浮游植物群落结构较复杂, 细胞密度变化显示时空的一致性, 优势种组成以及优势度存在明显的空间差异(P<0.05), 稻田的浅水环境以及小龙虾的养殖行为显著影响浮游植物群落的结构。鉴于藻类作为初级生产者对于水生态环境和小龙虾健康具有重要作用, 关注种养结合水体中藻类群落的演变规律对于保障稻田种养的综合效益具有积极意义。     

Characterization of remnant-like particles isolated by immunoaffinity gel from the plasma of type III and type IV hyperlipoproteinemic patients

Previous studies have investigated the potential atherogenicity and thrombogenicity of triglyceride-rich lipoprotein (TRL) remnants by isolating them from plasma within a remnant-like particle (RLP) fraction, using an immunoaffinity gel containing specific anti-apoB-100 and anti-apoA-I antibodies. In order to characterize lipoproteins in this RLP fraction and to determine to what extent their composition varies from one individual to another, we have used automated gel filtration chromatography to determine the size heterogeneity of RLP isolated from normolipidemic control subjects (n = 8), and from type III (n = 6) and type IV (n = 9) hyperlipoproteinemic patients, who by selection had similarly elevated levels of plasma triglyceride (406 +/- 43 and 397 +/- 35 mg/dl, respectively). Plasma RLP triglyceride, cholesterol, apoB, apoC-III, and apoE concentrations were elevated 2- to 6-fold (P < 0. 05) in hyperlipoproteinemic patients compared to controls. RLP fractions of type III patients were enriched in cholesterol and apoE compared to those of type IV patients, and RLP of type IV patients were enriched in triglyceride and apoC-III relative to those of normolipidemic subjects. In normolipidemic subjects, the majority of RLP had a size similar to LDL or HDL. The RLP of hyperlipoproteinemic patients were, however, larger and were similar in size to TRL, or were intermediate in size (i.e., ISL) between that of TRL and LDL. Compared to controls, ISL in the RLP fraction of type III patients were enriched in apoE relative to apoC-III, whereas in type IV patients they were enriched in apoC-III relative to apoE. These results demonstrate that: 1) RLP are heterogeneous in size and composition in both normolipidemic and hypertriglyceridemic subjects, and 2) the apoE and apoC-III composition of RLP is different in type III compared to type IV hyperlipoproteinemic patients.     

‘Only Fathers Smoking’ Contributes the Most to Socioeconomic Inequalities: Changes in Socioeconomic Inequalities in Infants’ Exposure to Second Hand Smoke over Time in Japan

BackgroundExposure to second hand smoke (SHS) is one of the major causes of premature death and disease among children. While socioeconomic inequalities exist for adult smoking, such evidence is limited for SHS exposure in children. Thus, this study examined changes over time in socioeconomic inequalities in infants’ SHS exposure in Japan.MethodsThis is a repeated cross-sectional study of 41,833 infants born in 2001 and 32,120 infants born in 2010 in Japan from nationally representative surveys using questionnaires. The prevalence of infants’ SHS exposure was determined and related to household income and parental education level. The magnitudes of income and educational inequalities in infants’ SHS exposure were estimated in 2001 and 2010 using both absolute and relative inequality indices.ResultsThe prevalence of SHS exposure in infants declined from 2001 to 2010. The relative index of inequality increased from 0.85 (95% confidence interval [CI], 0.80 to 0.89) to 1.47 (95% CI, 1.37 to 1.56) based on income and from 1.22 (95% CI, 1.17 to 1.26) to 2.09 (95% CI, 2.00 to 2.17) based on education. In contrast, the slope index of inequality decreased from 30.9 (95% CI, 29.3 to 32.6) to 20.1 (95% CI, 18.7 to 21.5) based on income and from 44.6 (95% CI, 43.1 to 46.2) to 28.7 (95% CI, 27.3 to 30.0) based on education. Having only a father who smoked indoors was a major contributor to absolute income inequality in infants’ SHS exposure in 2010, which increased in importance from 45.1% in 2001 to 67.0% in 2010.ConclusionsThe socioeconomic inequalities in infants’ second hand smoke exposure increased in relative terms but decreased in absolute terms from 2001 to 2010. Further efforts are needed to encourage parents to quit smoking and protect infants from second hand smoke exposure, especially in low socioeconomic households that include non-smoking mothers.     

Identification of juvenile hormone-active alkylphenols in the lobster Homarus americanus and in marine sediments

We have identified, by gas chromatography/mass spectrometry, four alkylphenols that are present in the hemolymph and tissues of the American lobster Homarus americanus and in marine sediments. These alkylphenols are used industrially in antioxidant formulations for plastic and rubber polymer manufacturing, and are similar in structure to a known endocrine disruptor, bisphenol A. The compound 2-t-butyl-4-(dimethylbenzyl)phenol was present at concentrations of 0.02 to 1.15 microg/ml in hemolymph and 8.95 to 21.58 microg/g in sediments. A second compound, 2,4-bis-(dimethylbenzyl)phenol, was present at concentrations between 0.07 and 19.78 microg/ml in hemolymph and 138.94 to 224.89 microg/g in sediment, while a third compound, 2,6-bis-(t-butyl)-4-(dimethylbenzyl)phenol, was found at concentrations between 0.01 and 13.00 microg/ml in hemolymph, 2.55 and 6.11 microg/g in hepatopancreas, and 47.85 and 74.66 microg/g in sediment. A fourth compound, 2,4-bis-(dimethylbenzyl)-6-t-butylphenol, was found at concentrations of 0.20 to 70.71 microg/ml in hemolymph, 23.56 to 26.89 microg/g in hepatopancreas, and 90.68 to 125.58 microg/g in sediment. These compounds, along with bisphenol A, 4-dimethylbenzylphenol, and nonylphenol, display high juvenile hormone activity in bioassays. Alkylphenols at high concentrations are toxic to crustaceans and may contribute significantly to lobster mortality; at lower concentrations, they are likely to have endocrine-disrupting effects.     

Community response grids: using information technology to help communities respond to bioterror emergencies

Access to accurate and trusted information is vital in preparing for, responding to, and recovering from an emergency. To facilitate response in large-scale emergency situations, Community Response Grids (CRGs) integrate Internet and mobile technologies to enable residents to report information, professional emergency responders to disseminate instructions, and residents to assist one another. CRGs use technology to help residents and professional emergency responders to work together in community response to emergencies, including bioterrorism events. In a time of increased danger from bioterrorist threats, the application of advanced information and communication technologies to community response is vital in confronting such threats. This article describes CRGs, their underlying concepts, development efforts, their relevance to biosecurity and bioterrorism, and future research issues in the use of technology to facilitate community response.     

Proteomic analysis of CA1 and CA3 regions of rat hippocampus and differential susceptibility to intermittent hypoxia

The CA1 and CA3 regions of the hippocampus markedly differ in their susceptibility to hypoxia in general, and more particularly to the intermittent hypoxia that characterizes sleep apnea. Proteomic approaches were used to identify proteins differentially expressed in the CA1 and CA3 regions of the rat hippocampus and to assess changes in protein expression following a 6-h exposure to intermittent hypoxia (IH). Ninety-nine proteins were identified, and 15 were differentially expressed in the CA1 and the CA3 regions. Following IH, 32 proteins in the CA1 region and only 7 proteins in the more resistant CA3 area were up-regulated. Hypoxia-regulated proteins in the CA1 region included structural proteins, proteins related to apoptosis, primarily chaperone proteins, and proteins involved in cellular metabolic pathways. We conclude that IH-mediated CA1 injury results from complex interactions between pathways involving increased metabolism, induction of stress-induced proteins and apoptosis, and, ultimately, disruption of structural proteins and cell integrity. These findings provide initial insights into mechanisms underlying differences in susceptibility to hypoxia in neural tissue, and may allow for future delineation of interventional strategies aiming to enhance neuronal adaptation to IH.     

Zinc response in pigeon pea as influenced by genotypic variability

Seven improved cultivars of pigeon pea (Cajanus cajan (L.) Mill sp.) were evaluated at 0 (original Zn deficient soil), 5 and 50 ppm Zn levels under greenhouse conditions. Plants were harvested at 6 weeks after sowing and at maturity. Responses to 5 ppm Zn in shoot at 6 weeks of growth, and in leaf, stem, pod-hull and grain at maturity ranged from 63 to 387, 37 to 116, 15 to 73,9 to 145 and 51 to 200%, respectively. Application of 50 ppm Zn in most of the cultivars did not markedly affect the yield of different plant parts. Zinc concentration at 0 Zn level in shoot at 6 weeks of growth and in leaf, stem, pod-hull and grain of different genotypes varied from 9.8 to 14.5, 13.7 to 21.2, 10.8 to 16.7, 4.17 to 5.83 and 9.2 to 16.7 ppm, respectively, and the increase in concentration with 5 ppm applied Zn ranged from 28 to 248, 28 to 89, 27 to 85, 20 to 142, and 105 to 254 per cent, respectively. The concentration further increased with an increase in Zn level to 50 ppm. There was less variation in the yield and tissue Zn concentration of different genotypes after Zn application. Phosphorus concentration at 0 Zn level in shoot at 6 weeks of growth, and in leaf, stem, pod-hull and grain of different genotypes varied from 0.50 to 0.71, 0.18 to 0.31, 0.11 to 0.24, 0.15 to 0.20 and 0.43 to 0.58% respectively. Zinc decreased P in all plant parts but relative decrease was more in vegetative parts than in grain. The variability in Zn response among pigeon pea genotypes could partly be attributed to the maintenance of proper P/Zn balance in metabolically active plant parts, such as, leaf, and partly to their capacity to exploit soil Zn and to translocate it to the above-ground parts.Contribution from the Department of Soils, Haryana Agricultural University, Hissar (India).     

北戴河朱鹮野化种群非繁殖期日间活动时间分配和行为节律

于2019年7月-2020年1月对北戴河朱鹮野化种群非繁殖期的日间活动时间分配和行为节律进行了分析。结果表明,休息（41.8%）、觅食（39.7%）和理羽（13.3%）是北戴河朱鹮非繁殖期的主要行为。在行为节律上,有3个觅食高峰、2个休息高峰和2个理羽高峰。成鸟和幼鸟行为分配相似,但成鸟的警戒和社群等其他行为比例显著较高。与野生种群相比,北戴河野化种群的觅食时间显著较少,主要原因是野生个体需要花费较多时间搜寻食物,且飞行等活动的能耗较高。朱鹮在冬季的休息和理羽行为比例高于夏秋季,觅食行为则相反,这样可以在低温和大风天气降低能耗,是对北戴河气候环境的有效适应。为了提高朱鹮对多种气候环境的适应能力,为今后北戴河朱鹮的再引入奠定基础,我们建议在冬季要确保朱鹮有充足的适宜觅食地,同时在野化网笼中进行必要的环境丰容,如在网笼内增加常绿树种和阔叶树种,部分侧网上安装防风板,或在网笼中设置防风避寒的伞棚等设施,以供朱鹮进行选择。     

Diabetes influences the effect of 17beta-estradiol on mechanical responses of rat urethra and detrusor strips

Estrogen deficiency is one of the factors involved in the stress incontinence in postmenopausal women, and estrogens have been used clinically in the treatment of urinary disorders during menopause. Sex hormones seem to be also involved in the diabetic changes of urinary bladder and urethra, because ovariectomy causes an increase in the micturition of streptozotocin-diabetic rats. In the present study diabetic and healthy female rats were used to investigate the effect of 17beta-estradiol on mechanical contractions to norepinephrine and to KCI and relaxations to ATP on isolated proximal urethral preparations as well as on contractions to ACh, ATP and KCl on detrusor smooth muscle strips. The data were compared with those obtained in OVX animals, with or without estradiol replacement. The present study showed that ovariectomy decreased the responses to ATP, NE and KCl in urethral preparations, and responses to ATP, ACh and KCl in bladder strips from both healthy and diabetic rats. Diabetes appeared to potentiate the effect of ovariectomy in both tissues. Estrogen replacement was able to recover functional responses in urethras of healthy rats. In diabetic rats, this treatment partially restored ATP-induced responses in both tissues, almost completely restored those to NE in urethra and those to ACh in bladder. This study clearly indicated that abnormalities of urethra and bladder function caused by ovariectomy can be restored by estrogen treatment also in diabetic animals, at least at an early stage of disease.     

Targeted Ablation of Nesprin 1 and Nesprin 2 from Murine Myocardium Results in Cardiomyopathy,Altered Nuclear Morphology and Inhibition of the Biomechanical Gene Response

Recent interest has focused on the importance of the nucleus and associated nucleoskeleton in regulating changes in cardiac gene expression in response to biomechanical load. Mutations in genes encoding proteins of the inner nuclear membrane and nucleoskeleton, which cause cardiomyopathy, also disrupt expression of a biomechanically responsive gene program. Furthermore, mutations in the outer nuclear membrane protein Nesprin 1 and 2 have been implicated in cardiomyopathy. Here, we identify for the first time a role for the outer nuclear membrane proteins, Nesprin 1 and Nesprin 2, in regulating gene expression in response to biomechanical load. Ablation of both Nesprin 1 and 2 in cardiomyocytes, but neither alone, resulted in early onset cardiomyopathy. Mutant cardiomyocytes exhibited altered nuclear positioning, shape, and chromatin positioning. Loss of Nesprin 1 or 2, or both, led to impairment of gene expression changes in response to biomechanical stimuli. These data suggest a model whereby biomechanical signals are communicated from proteins of the outer nuclear membrane, to the inner nuclear membrane and nucleoskeleton, to result in changes in gene expression required for adaptation of the cardiomyocyte to changes in biomechanical load, and give insights into etiologies underlying cardiomyopathy consequent to mutations in Nesprin 1 and 2.     

In vitro activity of MEKK2 and MEKK3 in detergents is a function of a valine to serine difference in the catalytic domain

MEKK2 and MEKK3 are mitogen-activated protein kinase kinase kinases (MAP3 kinases) of 70 and 71 kDa respectively that are markedly homologous (94%) in their kinase domains. Both MEKK2 and MEKK3 are able to activate the Jun kinase pathway in vivo. However, following routine immunoprecipitation in Triton X-100, MEKK2 but not MEKK3 is able to effectively phosphorylate both SEK-1 and MEK-1 and to undergo autophosphorylation. Unexpectedly, both MEKK2 and MEKK3 are functional in an in vitro kinase assay when cells are solubilized with the closely related detergent, NP-40. Given the high homology between these kinases, we set out to relate this differential sensitivity to Triton X-100 to differences in primary structure. A set of chimeric molecules were generated and the loss of activity in Triton X-100 mapped to kinase domain II/III and specifically to serine 390 of MEKK3 and valine 384 of MEKK2, residues immediately N-terminal to the active site lysine. Mutation of serine 390 of MEKK3 to a valine (as is found in MEKK2) conferred catalytic activity to MEKK3 in Triton X-100 whereas the reciprocal alteration of valine 384 of MEKK2 to a serine conferred lack of activity in Triton X-100 to MEKK2. Search of the protein database identified only three kinases, MEKK3, Pbs2p and Dd-PKI, with a serine or threonine at this site. The presence of a serine or threonine adjacent to the active site lysine in protein kinases is rare and, in MEKK3, results in detergent instability.     

Survival of Lactobacillus plantarum 44a after spraying and drying in feed and during exposure to gastrointestinal tract fluids in vitro

A good probiotic strain should be able to survive the conditions of handling and storage to be delivered in high concentration to the host. That is especially important when stressful conditions are prevalent in the carrier, for instance in low water content foods like animal feed. The aim of this research was to study the survival of the probiotic candidate Lactobacillus plantarum 44a after spraying and drying in feed, and during storage and exposure to gastrointestinal tract fluids in vitro. In addition, the viability of the strain during exposure to distilled water and 2% NaCl was studied. Feed was sprayed with a suspension of asymptotically equal to 2 x 10(10) CFU of L. plantarum 44a in 10, 15, 20, 25 and 30% v/w of the feed and dried to constant weight (6% moisture) in a convective oven at 25 degrees C. L. plantarum 44a survived 14.67, 36, 51.86, 78.9 and 105.3% respectively in relation to the original % v/w of the feed. After 3 weeks of storage at 25 degrees C, survival was similarly low in all the treatments. L. plantarum 44a stored in feed containing 13% moisture, vacuum-packaged and stored in refrigeration, maintained high viability (approximately 100%) after 1 year of storage. Survival was not affected after feed-containing lactobacilli was exposed to gastrointestinal fluids in a simulation model. Viability of L. plantarum 44a as a cell suspension in PBS added directly to distilled water or distilled water with 2% NaCl was maintained up to 48 h; after 72 h, viability started to decline. It is concluded that L. plantarum 44a maintained high viability after being dried and stored in feed even after exposure to gastric and intestinal fluids in vitro.