Terminalia pallida fruit ethanolic extract ameliorates lipids,lipoproteins, lipid metabolism marker enzymes and paraoxonase in isoproterenol-induced myocardial infarcted rats

The present study aimed to evaluate the effect of Terminalia pallida fruit ethanolic extract (TpFE) on lipids, lipoproteins, lipid metabolism marker enzymes and paraoxonase (PON) in isoproterenol (ISO)-induced myocardial infarcted rats. PON is an excellent serum antioxidant enzyme which involves in the protection of low density lipoprotein cholesterol (LDL-C) from the process of oxidation for the prevention of cardiovascular diseases. ISO caused a significant increase in the concentration of total cholesterol, triglycerides, LDL-C, very low density lipoprotein cholesterol and lipid peroxidation whereas significant decrease in the concentration of high density lipoprotein cholesterol. ISO administration also significantly decreased the activities of lecithin cholesterol acyl transferase, PON and lipoprotein lipase whereas significantly increased the activity of 3-hydroxy-3-methylglutaryl-coenzyme-A reductase. Oral pretreatment of TpFE at doses 100, 300 and 500?mg/kg body weight (bw) and gallic acid (15?mg/kg bw) for 30?days challenged with concurrent injection of ISO (85?mg/kg bw) on 29th and 30th day significantly attenuated these alterations and restored the levels of lipids, lipoproteins and the activities of lipid metabolizing enzymes. Also TpFE significantly elevated the serum antioxidant enzyme PON. This is the first report revealed that pretreatment with TPFE ameliorated lipid metabolic marker enzymes and increased the antioxidant PON in ISO treated male albino Wistar rats.     

A Synthetic Lethal Screen Identifies DNA Repair Pathways that Sensitize Cancer Cells to Combined ATR Inhibition and Cisplatin Treatments

The DNA damage response kinase ATR may be a useful cancer therapeutic target. ATR inhibition synergizes with loss of ERCC1, ATM, XRCC1 and DNA damaging chemotherapy agents. Clinical trials have begun using ATR inhibitors in combination with cisplatin. Here we report the first synthetic lethality screen with a combination treatment of an ATR inhibitor (ATRi) and cisplatin. Combination treatment with ATRi/cisplatin is synthetically lethal with loss of the TLS polymerase ζ and 53BP1. Other DNA repair pathways including homologous recombination and mismatch repair do not exhibit synthetic lethal interactions with ATRi/cisplatin, even though loss of some of these repair pathways sensitizes cells to cisplatin as a single-agent. We also report that ATRi strongly synergizes with PARP inhibition, even in homologous recombination-proficient backgrounds. Lastly, ATR inhibitors were able to resensitize cisplatin-resistant cell lines to cisplatin. These data provide a comprehensive analysis of DNA repair pathways that exhibit synthetic lethality with ATR inhibitors when combined with cisplatin chemotherapy, and will help guide patient selection strategies as ATR inhibitors progress into the cancer clinic.     

PtrWRKY51基因的克隆及抗旱表达特性分析

为明确毛果杨WRKY家族成员PtrWRKY51基因功能,以Nisqually-1株系毛果杨为模板,克隆得到PtrWRKY51基因CDS序列。通过生物信息学分析,结合酵母自激活验证、亚细胞定位及模拟干旱胁迫下的实时荧光定量PCR(qRT-PCR)对PtrWRKY51基因功能进行初步研究。结果表明：PtrWRKY51全长579 bp,编码192 aa。生物信息学分析及亚细胞定位试验结果表明,PtrWRKY51蛋白为非跨膜碱性不稳定亲水蛋白,定位于细胞核,含有WRKY家族特有的保守结构域,是第IIc类WRKY转录因子;酵母自激活验证试验表明PtrWRKY51基因具有自激活活性;qRT-PCR分析表明,8%PEG6000模拟干旱胁迫下,该基因在胁迫12 h后茎部与叶部相对表达量达到最大值,根部则出现在24 h,研究可为PtrWRKY51抗逆及生物学功能进一步研究提供参考。     

ATR and ATRIP Are Recruited to Herpes Simplex Virus Type 1 Replication Compartments Even though ATR Signaling Is Disabled

Although the herpes simplex virus type 1 (HSV-1) genome might be expected to induce a DNA damage response, the ATR kinase is not activated in infected cells. We previously proposed that spatial uncoupling of ATR from its interaction partner, ATRIP, could be the basis for inactivation of the ATR kinase in infected cells; however, we now show that ATR and ATRIP are in fact both recruited to HSV-1 replication compartments and can be coimmunoprecipitated from infected-cell lysates. ATRIP and replication protein A (RPA) are recruited to the earliest detectable prereplicative sites, stage II microfoci. In a normal cellular DNA damage response, ATR/ATRIP are recruited to stretches of RPA-coated single-stranded DNA in an RPA- and kinase-dependent manner, resulting in the phosphorylation of RPA by ATR in damage foci. In contrast, in HSV-1-infected cells, RPA is not phosphorylated, and endogenous phosphorylated RPA is excluded from stage II microfoci; in addition, the recruitment of ATR/ATRIP is independent of RPA and the kinase activity of ATR. Furthermore, we show that ATR/ATRIP play a beneficial role in viral gene expression and virus production. Although ICP0 has been shown to be important for partial inactivation of other cellular DNA repair pathways, we show that ICP0 is not responsible for the inactivation of ATR signaling and, furthermore, that neither ATR nor ATRIP is a target of ICP0 degradation. Thus, ATR and ATRIP may function outside the context of the canonical ATR damage signaling pathway during HSV-1 infection to participate in the viral life cycle.Herpes simplex virus type 1 (HSV-1) is a large linear double-stranded DNA virus that replicates in the nucleus of the host cell. The incoming viral genome contains nicks and gaps (42), and cellular DNA repair machinery might be expected to recognize it as damaged, resulting in the activation of one or more cellular DNA damage pathways. Activation of DNA damage response pathways can result not only in repair of the damaged DNA but also in cell cycle arrest, gene silencing, and apoptosis (9). The later outcomes could result in suppression of viral gene expression and DNA replication and thus have negative consequences for lytic infection. Activation of a cellular DNA damage response during viral infection could, therefore, represent a form of intrinsic antiviral immunity (14, 15). On the other hand, HSV-1 and other DNA viruses which replicate in the nucleus have also been shown to utilize cellular DNA repair machinery to promote productive infection (28). Thus, HSV-1 has apparently evolved to manipulate the host DNA damage response by utilizing some components and inactivating others in an attempt to create an environment conducive to lytic viral infection.The cellular DNA damage response is regulated by the three phosphoinositide 3-kinase-related kinases (PIKKs), DNA-PK (DNA-dependent protein kinase), ATM (ataxia-telangiectasia-mutated), and ATR (ATM and Rad3-related) (1, 9). DNA-PK and ATM respond predominantly to double-strand breaks, and ATR responds to stalled replication forks and long stretches of single-stranded DNA (ssDNA). DNA-PK is required for nonhomologous end joining (NHEJ), while ATM activation promotes homologous recombination. Interestingly, in some cell types, the catalytic subunit of DNA-PK (DNA-PKcs) is proteolytically degraded during infection by the immediate-early (IE) protein ICP0, a viral E3 ubiquitin ligase (25, 37), thereby resulting in the probable inactivation of the NHEJ pathway. ATM kinase activity, on the other hand, is activated during HSV-1 infection once viral DNA replication is initiated (26, 47, 56). Despite phosphorylation of several ATM targets, ATM signaling is also modulated by ICP0, which degrades the ubiquitin ligases RNF8 and RNF168. The function of these ubiquitin ligases is to promote the tethering of ATM pathway proteins at sites of cellular DNA damage (27). Thus, ICP0 functions to partially inactivate portions of both the DNA-PK- and ATM-mediated repair pathways.During a cellular DNA damage response, ATM activation and processing of DNA ends generate ssDNA adjacent to double-stranded DNA (dsDNA), a structure that is known to activate ATR (9, 38). The ssDNA is coated by the cellular ssDNA binding protein, replication protein A (RPA), which then serves to recruit ATR through a direct interaction with ATR-interacting protein (ATRIP) (4, 12, 58). ATR signaling results in the phosphorylation of many substrates, including RPA and Chk1. During HSV-1 infection, the ATR substrates RPA and Chk1 are not phosphorylated (47, 54-56), indicating that ATR signaling may be disabled.A hallmark of HSV-1 infection is the reorganization of the infected-cell nucleus, resulting in the formation of large globular replication compartments as well as the rearrangement of cellular proteins involved in several homeostatic pathways. In addition to cellular DNA repair proteins, HSV-1 infection also causes the reorganization of components of the cellular protein quality control pathways, resulting in the formation of virus-induced chaperone-enriched (VICE) domains, which act to maintain nuclear protein quality control during infection (31). Viral gene expression, DNA replication, and encapsidation of viral genomes occur in replication compartments (24, 39, 41). In this work we revisit the study of proteins recruited to and restricted from replication compartments in an attempt to better understand how HSV-1 manipulates components of the cellular DNA damage response for its own benefit.     

紫毛香茶菜中的黄酮类化合物

从紫毛香茶菜（Isodon enanderianus)的茎叶提取物中分离到7个黄酮类化合物,经波谱分析鉴定,其中一个为新的黄酮甙,即5-羟基-6,7,4-三甲氧基黄酮-8-O-β-D-葡萄糖甙（1）;其它6个已知的黄酮类化合物分别为芫花素（genkwanin,2)、滨蓟素（cirsimar-tin,3)、胡麻素（pedalitin,4)、鼬瓣花亭（ladanetin,5)、isothymusin-8-O-β-D-glucoside（6）和槲皮甙（quercitrin,7)。     

A Dutch guideline for the treatment of scoliosis in neuromuscular disorders

Background

Children with neuromuscular disorders with a progressive muscle weakness such as Duchenne Muscular Dystrophy and Spinal Muscular Atrophy frequently develop a progressive scoliosis. A severe scoliosis compromises respiratory function and makes sitting more difficult. Spinal surgery is considered the primary treatment option for correcting severe scoliosis in neuromuscular disorders. Surgery in this population requires a multidisciplinary approach, careful planning, dedicated surgical procedures, and specialized after care.

Methods

The guideline is based on scientific evidence and expert opinions. A multidisciplinary working group representing experts from all relevant specialties performed the research. A literature search was conducted to collect scientific evidence in answer to specific questions posed by the working group. Literature was classified according to the level of evidence.

Results

For most aspects of the treatment scientific evidence is scarce and only low level cohort studies were found. Nevertheless, a high degree of consensus was reached about the management of patients with scoliosis in neuromuscular disorders. This was translated into a set of recommendations, which are now officially accepted as a general guideline in the Netherlands.

Conclusion

In order to optimize the treatment for scoliosis in neuromuscular disorders a Dutch guideline has been composed. This evidence-based, multidisciplinary guideline addresses conservative treatment, the preoperative, perioperative, and postoperative care of scoliosis in neuromuscular disorders.     

A Novel CMKLR1 Small Molecule Antagonist Suppresses CNS Autoimmune Inflammatory Disease

Therapies that target leukocyte trafficking pathways can reduce disease activity and improve clinical outcomes in multiple sclerosis (MS). Experimental autoimmune encephalomyelitis (EAE) is a widely studied animal model that shares many clinical and histological features with MS. Chemokine-like receptor-¹ (CMKLR1) is a chemoattractant receptor that is expressed by key effector cells in EAE and MS, including macrophages, subsets of dendritic cells, natural killer cells and microglia. We previously showed that CMKLR1-deficient (CMKLR1 KO) mice develop less severe clinical and histological EAE than wild-type mice. In this study, we sought to identify CMKLR1 inhibitors that would pharmaceutically recapitulate the CMKLR1 KO phenotype in EAE. We identified 2-(α-naphthoyl) ethyltrimethylammonium iodide (α-NETA) as a CMKLR1 small molecule antagonist that inhibits chemerin-stimulated β-arrestin2 association with CMKLR1, as well as chemerin-triggered CMKLR1⁺ cell migration. α-NETA significantly delayed the onset of EAE induced in C57BL/6 mice by both active immunization with myelin oligodendrocyte glycoprotein peptide 35-55 and by adoptive transfer of encephalitogenic T cells. In addition, α-NETA treatment significantly reduced mononuclear cell infiltrates within the CNS. This study provides additional proof-of-concept data that targeting CMKLR1:chemerin interactions may be beneficial in preventing or treating MS.     

Survey of aflatoxin M1 in cows’ milk from free-grazing cows in Abeokuta,Nigeria

Aflatoxin M₁ (AFM₁) in milk from 100 different herds of free-grazing cows in Abeokuta, Nigeria, was analysed by immunoaffinity column cleanup and HPLC with fluorescence detection. AFM₁ was found in 75 % of the samples, the toxin levels in positive samples ranged from 9.0 to 456.0 ng/l. The mean AFM₁ level in positive samples was 108.15 ng/l, exceeding, for example, the European Union maximum level by a factor of two. These results indicated that there is an urgent need to more closely control the milk of free-grazing cows for AFM₁ in order to protect the health of humans consuming milk and milk products.