Astragaloside IV Protects against Isoproterenol-Induced Cardiac Hypertrophy by Regulating NF-κB/PGC-1α Signaling Mediated Energy Biosynthesis

We previously reported that Astragaloside IV (ASIV), a major active constituent of Astragalus membranaceus (Fisch) Bge protects against cardiac hypertrophy in rats induced by isoproterenol (Iso), however the mechanism underlying the protection remains unknown. Dysfunction of cardiac energy biosynthesis contributes to the hypertrophy and Nuclear Factor κB (NF-κB)/Peroxisome Proliferator-Activated Receptor-γ Coactivator 1α (PGC-1α) signaling gets involved in the dysfunction. The present study was designed to investigate the mechanism by which ASIV improves the cardiac hypertrophy with focuses on the NF-κB/PGC-1α signaling mediated energy biosynthesis. Sprague-Dawley (SD) rats or Neonatal Rat Ventricular Myocytes (NRVMs) were treated with Iso alone or in combination with ASIV. The results showed that combination with ASIV significantly attenuated the pathological changes, reduced the ratios of heart weight/body weight and Left ventricular weight/body weight, improved the cardiac hemodynamics, down-regulated mRNA expression of Atrial Natriuretic Peptide (ANP) and Brain Natriuretic Peptide (BNP), increased the ratio of ATP/AMP, and decreased the content of Free Fat Acid (FFA) in heart tissue of rats compared with Iso alone. In addition, pretreatment with ASIV significantly decreased the surface area and protein content, down-regulated mRNA expression of ANP and BNP, increased the ratio of ATP/AMP, and decreased the content of FFA in NRVMs compared with Iso alone. Furthermore, ASIV increased the protein expression of ATP5D, subunit of ATP synthase and PGC-1α, inhibited translocation of p65, subunit of NF-κB into nuclear fraction in both rats and NRVMs compared with Iso alone. Parthenolide (Par), the specific inhibitor of p65, exerted similar effects as ASIV in NRVMs. Knockdown of p65 with siRNA decreased the surface areas and increased PGC-1α expression of NRVMs compared with Iso alone. The results suggested that ASIV protects against Iso-induced cardiac hypertrophy through regulating NF-κB/PGC-1α signaling mediated energy biosynthesis.     

High Prevalence of Vancomycin-Resistant Enterococci in Swedish Sewage

In Europe the use of the growth promoter avoparcin is considered to have selected for vancomycin-resistant enterococci (VRE). Sweden ceased using avoparcin in 1986, and only occasional cases of VRE from hospitals have been reported since 1995. Within the framework of a European study, samples from urban raw sewage, treated sewage, surface water, and hospital sewage in Sweden (n = 118) were screened for VRE. Surprisingly, VRE were isolated from 21 of 35 untreated sewage samples (60%), from 5 of 14 hospital sewage samples (36%), from 6 of 32 treated sewage samples (19%), and from 1 of 37 surface water samples. Thirty-five isolates from 33 samples were further characterized by geno- and phenotyping, MIC determination, and PCR analysis. Most isolates (30 of 35) carried the vanA gene, and the majority (24 of 35) of the isolates were Enterococcus faecium. Most of the VRE were multiresistant. The typing revealed high diversity of the isolates. However, one major cluster with seven identical or similar isolates was found. These isolates came from three different sewage treatment plants and were collected at different occasions during 1 year. All VRE from hospital sewage originated from one of the two hospitals studied. That hospital also had vancomycin consumption that was 10-fold that of the other. We conclude that VRE were commonly found in sewage samples in Sweden. The origin might be both healthy individuals and individuals in hospitals. Possibly, antimicrobial drugs or chemicals released into the sewage system may sustain VRE in the system.     

Root distribution and water uptake patterns of maize cultivars field-grown under differential irrigation

Summary Rooting and water uptake patterns were determined for three maize (Zea mays L) varieties field-grown during the 1983/84 dry season under seven irrigation levels on a sandy loam soil. Roots were mainly concentrated in the top 22 cm due to a 40 cm thick compact gravelly layer occurring from about this depth in the profile. There were significant varietal differences, distinguished by root length density (RLD) and length/weight ratio (LAR) distributions at depth and at varying soil moisture regimes. These properties were related to water extraction patterns and grain yields. Yields obtained at adequate soil moisture were 6.9 tha⁻¹ for TZESR-W (var 1), 4.2 t/ha for TZSR-W (var 2) and 3.7t ha⁻¹ for FARZ-7 (var 3). These yeilds were respectively associated with maximum RLD of 2.56, 1.88 and 1.70 cm cm⁻³ and corresponding LWR of 2.64, 1.93 and 1.62 cm mg⁻¹. Average seasonal water uptake was estimated at 4.2, 3.0 and 2.8 mm day⁻¹ for var 1, 2 and 3, respectively. Better performance of var 1 was attributed to the development of a more active and deep rooting system.     

小鼠pEGFP-Hoxa11真核表达载体的构建及表达

目的 构建和鉴定Hoxa11和EGFP双基因共表达真核载体.方法 采用DNA重组技术,将目的 基因Hoxa11克隆至含有报告基因EGFP的pEGFP-N1真核表达载体中,构建的真核表达载体pEGFP-Hoxa11经PCR,双酶切及基因测序鉴定;转染至CHO细胞,荧光显微镜下观察重组质粒的表达,提取细胞蛋白Western印迹检测蛋白表达.结果 pEGFP-Hoxa11重组质粒构建成功.构建的真核表达载体pEGFP-Hoxa11能在CHO细胞中有效表达.结论 成功构建了共表达Hoxa11和EGFP的真核表达载体,并能在CHO细胞中有效表达.为进一步研究Hoxa11的功能提供实验基础.     

Substrate binding and translocation of the serotonin transporter studied by docking and molecular dynamics simulations

The serotonin (5-HT) transporter (SERT) plays an important role in the termination of 5-HT-mediated neurotransmission by transporting 5-HT away from the synaptic cleft and into the presynaptic neuron. In addition, SERT is the main target for antidepressant drugs, including the selective serotonin reuptake inhibitors (SSRIs). The three-dimensional (3D) structure of SERT has not yet been determined, and little is known about the molecular mechanisms of substrate binding and transport, though such information is very important for the development of new antidepressant drugs. In this study, a homology model of SERT was constructed based on the 3D structure of a prokaryotic homologous leucine transporter (LeuT) (PDB id: 2A65). Eleven tryptamine derivates (including 5-HT) and the SSRI (S)-citalopram were docked into the putative substrate binding site, and two possible binding modes of the ligands were found. To study the conformational effect that ligand binding may have on SERT, two SERT–5-HT and two SERT–(S)-citalopram complexes, as well as the SERT apo structure, were embedded in POPC lipid bilayers and comparative molecular dynamics (MD) simulations were performed. Our results show that 5-HT in the SERT–5-HT^B complex induced larger conformational changes in the cytoplasmic parts of the transmembrane helices of SERT than any of the other ligands. Based on these results, we suggest that the formation and breakage of ionic interactions with amino acids in transmembrane helices 6 and 8 and intracellular loop 1 may be of importance for substrate translocation.     

Site-specific phosphorylation of Tau protein is associated with deacetylation of microtubules in mouse spermatogenic cells during meiosis

Tau is one of the microtubule-associated proteins and a major component of paired helical filaments, a hallmark of Alzheimer’s disease. Its expression has also been indicated in the testis. However, its function and modification in the testis have not been established. Here, we analyzed the dynamics of phosphorylation patterns during spermatogenesis. The expression of Tau protein and its phosphorylation were shown in the mouse testis. Immunohistochemistry revealed that the phosphorylation was strongly detected during meiosis. Correspondingly, the expression of acetylated tubulin was inversely weakened during meiosis. These results suggest that phosphorylation of Tau protein contributes to spermatogenesis, especially in meiosis.     

High Prevalence,Coinfection Rate,and Genetic Diversity of Retroviruses in Wild Red Colobus Monkeys (Piliocolobus badius badius) in Ta? National Park,C?te d'Ivoire

Simian retroviruses are precursors of all human retroviral pathogens. However, little is known about the prevalence and coinfection rates or the genetic diversity of major retroviruses—simian immunodeficiency virus (SIV), simian T-cell lymphotropic virus type 1 (STLV-1), and simian foamy virus (SFV)—in wild populations of nonhuman primates. Such information would contribute to the understanding of the natural history of retroviruses in various host species. Here, we estimate these parameters for wild West African red colobus monkeys (Piliocolobus badius badius) in the Taï National Park, Côte d''Ivoire. We collected samples from a total of 54 red colobus monkeys; samples consisted of blood and/or internal organs from 22 monkeys and additionally muscle and other tissue samples from another 32 monkeys. PCR analyses revealed a high prevalence of SIV, STLV-1, and SFV in this population, with rates of 82%, 50%, and 86%, respectively. Forty-five percent of the monkeys were coinfected with all three viruses while another 32% were coinfected with SIV in combination with either STLV or SFV. As expected, phylogenetic analyses showed a host-specific pattern for SIV and SFV strains. In contrast, STLV-1 strains appeared to be distributed in genetically distinct and distant clades, which are unique to the Taï forest and include strains previously described from wild chimpanzees in the same area. The high prevalence of all three retroviral infections in P. b. badius represents a source of infection to chimpanzees and possibly to humans, who hunt them.Lentiviruses and deltaretroviruses that infect African nonhuman primates have received considerable attention as they are the precursors of all pathogenic human retroviruses: human immunodeficiency virus types 1 and 2 (HIV-1/HIV-2) and human T-cell lymphotropic virus type 1 (HTLV-1). These human infections are the results of past zoonotic transfers of simian immunodeficiency virus (SIV) and simian T-cell lymphotropic viruses type 1 (STLV-1) from wild monkeys and apes into local human populations, presumably through primate hunting and handling of primate bushmeat (13, 19, 43, 46, 55, 58, 59). Via the same route, zoonotic transmission of simian foamy virus (SFV), a spumaretrovirus whose exact pathogenicity in human hosts is still unknown, has also been shown (64). The increasing contact between humans and wild primates implies that further zoonotic transmission of retroviruses is likely to happen (42, 63). Studying the occurrence and circulation of simian retroviruses such as SIV, STLV-1, and SFV in wild primate populations enables us to better understand retrovirus evolution in primates and also provides tools for monitoring possible future retroviral zoonotic events.Systematic studies of SIV, STLV-1, and SFV in wild primates are relatively rare. Many use bushmeat samples, which can vary in their quality and are prone to cross-contamination from butchering and storage with other carcasses. Confiscated primates are also not representative of the situation in the wild since the animals are caught at a young age when the occurrence of different retroviruses may be extremely low (24). The technical possibilities for the detection of various pathogens in noninvasive samples such as urine and feces have greatly improved and are frequently used; however, in general, the sensitivity of detection methods is higher when blood and tissue samples are used (25, 32, 47). Such samples can be collected if fresh carcasses are found, or they can be collected by anesthetizing live primates for sampling purpose, animal translocation, or medical intervention, such as snare removal. The practical and ethical issues of each of the sampling methods have been discussed elsewhere (12, 14).Red colobus monkeys [Procolobus (Piliocolobus)] are interesting subjects for retroviral infection studies for a number of reasons. First, they are widely distributed (yet in a fragmented manner) from East to West Africa, which suggests that red colobus species and subspecies, or more likely ancestor(s) of these, could have been key hosts in transmitting retroviruses across tropical Africa (4, 54). Second, as they are herbivore primates, the hunting of other primates can be excluded as a route of infection. Finally, these monkeys are frequently hunted by humans and chimpanzees and represent a possibly large reservoir for retroviruses and other pathogens that ought to be investigated further (2, 45).Very little information is available about the prevalence and coinfection of SIV, STLV-1, and SFV in wild red colobus monkeys across Africa. In other colobine monkeys only SIV has been documented: in olive colobus (Procolobus verus) in Côte d''Ivoire and in black and white colobus (Colobus guereza) in Cameroon (7, 8). Based on fecal samples from habituated adult individuals, the prevalence of SIV in West African red colobus monkeys (SIVwrc; local subspecies, Piliocolobus badius badius) has been estimated to a minimum of 26% in the Taï National Park, Côte d''Ivoire, but the authors recognized the low sensitivity of viral RNA detection in fecal samples (34). Another study conducted on the same population revealed that 5 out of 10 blood samples were SIV positive (7). These results highlight that the most reliable prevalence data are based on analyses of blood/tissue samples although such sampling is not always feasible for reasons discussed above. Published prevalence information concerning STLV-1 and SFV in wild red colobus monkeys (STLV-1wrc and SFVwrc) in the same area is restricted to results obtained from analyses of a limited number of blood and necropsy samples collected as a part of studies whose focus was on cross-species transmission of these two viruses to chimpanzees (27, 28). However, these samples indicated a high prevalence of STLV-1wrc and SFVwrc in the red colobus monkey population (56% and 90%, respectively). A recent study from Uganda, East Africa, estimated the prevalence of SIV, STLV-1, and SFV in another red colobus species (Piliocolobus rufomitratus tephrosceles) to be 22.6%, 6.4%, and 97%, respectively (15). The study was performed using blood samples collected from anesthetized wild red colobus monkeys living in their natural habitat, which allowed reliable assessment of the prevalence and genetic diversity of these three retroviruses.The preliminary data from the Taï National Park indicate that there might be great variation in the prevalence of retroviruses across the African continent, even in closely related species of wild primates. Here, we aimed at generating reliable prevalence and coinfection data for SIVwrc, STLV-1wrc, and SFVwrc based on the analysis of blood and tissue samples from wild Western red colobus monkeys. We expected that this would allow for proper comparison of retroviral prevalence in the allied species P. b. badius and P. r. tephrosceles.     

High prevalence of vancomycin-resistant enterococci in Swedish sewage

In Europe the use of the growth promoter avoparcin is considered to have selected for vancomycin-resistant enterococci (VRE). Sweden ceased using avoparcin in 1986, and only occasional cases of VRE from hospitals have been reported since 1995. Within the framework of a European study, samples from urban raw sewage, treated sewage, surface water, and hospital sewage in Sweden (n = 118) were screened for VRE. Surprisingly, VRE were isolated from 21 of 35 untreated sewage samples (60%), from 5 of 14 hospital sewage samples (36%), from 6 of 32 treated sewage samples (19%), and from 1 of 37 surface water samples. Thirty-five isolates from 33 samples were further characterized by geno- and phenotyping, MIC determination, and PCR analysis. Most isolates (30 of 35) carried the vanA gene, and the majority (24 of 35) of the isolates were Enterococcus faecium. Most of the VRE were multiresistant. The typing revealed high diversity of the isolates. However, one major cluster with seven identical or similar isolates was found. These isolates came from three different sewage treatment plants and were collected at different occasions during 1 year. All VRE from hospital sewage originated from one of the two hospitals studied. That hospital also had vancomycin consumption that was 10-fold that of the other. We conclude that VRE were commonly found in sewage samples in Sweden. The origin might be both healthy individuals and individuals in hospitals. Possibly, antimicrobial drugs or chemicals released into the sewage system may sustain VRE in the system.