Zymosan-mediated inflammation impairs in vivo reverse cholesterol transport

Inflammation has been proposed to impair HDL function and reverse cholesterol transport (RCT). We investigated the effects of inflammation mediated by zymosan, a yeast glucan, on multiple steps along the RCT pathway in vivo and ex vivo. Acute inflammation with 70 mg/kg zymosan impaired RCT to plasma, liver, and feces similarly by 17-22% (P < 0.05), with no additional block at the liver. Hepatic gene expression further demonstrated no change in ABCG5, ABCB4, and ABCB11 expression but a decline in ABCG8 mRNA (32% P < 0.05). Plasma from zymosan-treated mice had a 21% decrease in cholesterol acceptor ability (P < 0.01) and a 35% decrease in ABCA1-specific efflux capacity (P < 0.01) in vitro. Zymosan treatment also decreased HDL levels and led to HDL remodeling with increased incorporation of serum amyloid A. In addition, cholesterol efflux from cultured macrophages declined with zymosan treatment in a dose dependent manner. Taken together, our results suggest that zymosan impairs in vivo RCT primarily by decreasing macrophage-derived cholesterol entering the plasma, with minimal additional blocks downstream. Our study supports the notion that RCT impairment is one of the mechanisms for the increased atherosclerotic burden observed in inflammatory conditions.     

Dual recognition-incision enzymes might be involved in mismatch repair and meiosis

Mismatch repair in many organisms depends on three proteins: the mismatch-recognition protein MutS, a nicking endonuclease MutH, and MutL, which acts as a scaffold between these. However, many genomes lack MutL but possess MutS. In one of these cases, in a coral mitochondrial genome, a gene is present that encodes a MutS protein fused to an HNH nicking endonuclease, potentially eliminating the requirement for MutL. Likewise, many prokaryotes could operate similarly, independently of MutL by encoding a fused MutS-Smr (MutS2) protein. Smr, which is proposed to be a nicking endonuclease, can also be found separately in many eukaryotes, where it might play a role in mismatch repair or meiotic chromosome crossing-over.     

Proteomic analysis of differentially expressed serum proteins in lung cancer

Lung cancer continues to represent a major public health concern with high morbidity and mortality worldwide. Early detection of lung cancer is problematic due to a lack of diagnostic markers with high sensitivity and specificity. To determine the differently expressed proteins in the serum of lung cancer and identify the function of such proteins, two-dimensional electrophoresis (2DE) and liquid chromatography mass spectrometry (LC-MS) were used to screen the serum of lung cancer model induced by 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK). A total of 25 protein spots were qualitatively different and 6 were quantitatively different in the serum from rats bearing induced lung cancer when compared with normal controls. Two of the proteins that showed major changes in concentration in sera were identified to be Immunoglobulin γ 2A chain C region (heavy chain) and Transferrin by LC-MS/MS.     

Transfer of the barrier to radial oxygen loss in roots of Hordeum marinum to wheat (Triticum aestivum): evaluation of four H. marinum-wheat amphiploids

? Wide hybridization of waterlogging-tolerant Hordeum marinum with wheat (Triticum aestivum) to produce an amphiploid might be one approach to improve waterlogging tolerance in wheat. ? Growth, root aerenchyma and porosity, and radial oxygen loss (ROL) along roots were measured in four H. marinum-wheat amphiploids and their parents (four accessions of H. marinum and Chinese Spring wheat) in aerated or stagnant nutrient solution. A soil experiment was also conducted. ? Hordeum marinum maintained shoot dry mass in stagnant nutrient solution, whereas the growth of wheat was markedly reduced (40% of aerated control). Two of the four amphiploids were more tolerant than wheat (shoot dry masses of 59-72% of aerated controls). The porosity of adventitious roots when in stagnant solution was higher in H. marinum (19-25%) and the four amphiploids (20-24%) than in wheat (16%). In stagnant solution, adventitious roots of H. marinum formed a strong ROL barrier in basal zones, whereas, in wheat, the barrier was weak. Two amphiploids formed a strong ROL barrier and two formed a moderate barrier when in stagnant solution. ? This study demonstrates the transfer of higher root porosity and a barrier to ROL from H. marinum to wheat through wide hybridization and the production of H. marinum-wheat amphiploids.     

Re-emergence of glanders in India — Report of Maharashtra state

Glanders, a notifiable highly contagious disease primarily of equids, is a disease of high zoonotic importance. Caused by gram-negative bacillus, Burkholderia mallei, the disease was restricted to certain pockets of India with sporadic cases. Recently, a major outbreak of glanders occurred in India starting from Maharashtra. Following clinical signs and symptoms and laboratory investigations on serum, nasal swab and pus swab samples, it was confirmed as glanders among equines in Pune and Panchgani areas of Maharashtra. One pus sample and three nasal swabs yielded B. mallei isolates while 23 serum samples were found positive for glanders by complement fixation test (CFT). The disease was successfully controlled in the state by following strategies for prevention of spread of the disease to other areas in accordance with Glanders and Farcy Act, 1899. Follow up of the occurrence in Maharashtra revealed negative status based on testing and physical surveillance on more than 3,500 equines thereafter. Investigations indicated that the nidus of infection may be present elsewhere in North India.     

Advances in molecular detection of Aspergillus: an update

Filamentous cosmopolitan fungi of the genus Aspergillus can be harmful in two ways, directly they can be opportunistic pathogens causing aspergillosis and indirectly due to aflatoxin production on food products which can lead to aflatoxicosis. Therefore, a number of methods have been proposed so far for detection of the fungi with lowest possible concentration at the earliest. Molecular methods such as PCR and/or in combination with certain techniques have been found to be useful for Aspergillus detection. We discuss here various technologies that have emerged in recent years and can possibly be used for the molecular detection of Aspergillus in an efficient way. These methods like RSIC, C-probe, and inversion probe with pyrosequencing or direct ss/dsDNA detection have been used for the identification of fungal or bacterial pathogens and thus formulate a ‘gold standard’ for Aspergillus detection.     

Phagocytosis Is the Main CR3-Mediated Function Affected by the Lupus-Associated Variant of CD11b in Human Myeloid Cells

The CD11b/CD18 integrin (complement receptor 3, CR3) is a surface receptor on monocytes, neutrophils, macrophages and dendritic cells that plays a crucial role in several immunological processes including leukocyte extravasation and phagocytosis. The minor allele of a non-synonymous CR3 polymorphism (rs1143679, conversation of arginine to histidine at position 77: R77H) represents one of the strongest genetic risk factor in human systemic lupus erythematosus, with heterozygosity (77R/H) being the most common disease associated genotype. Homozygosity for the 77H allele has been reported to reduce adhesion and phagocytosis in human monocytes and monocyte-derived macrophages, respectively, without affecting surface expression of CD11b. Herein we comprehensively assessed the influence of R77H on different CR3-mediated activities in monocytes, neutrophils, macrophages and dendritic cells. R77H did not alter surface expression of CD11b including its active form in any of these cell types. Using two different iC3b-coated targets we found that the uptake by heterozygous 77R/H macrophages, monocytes and neutrophils was significantly reduced compared to 77R/R cells. Allele-specific transduced immortalized macrophage cell lines demonstrated that the minor allele, 77H, was responsible for the impaired phagocytosis. R77H did not affect neutrophil adhesion, neutrophil transmigration in vivo or Toll-like receptor 7/8-mediated cytokine release by monocytes or dendritic cells with or without CR3 pre-engagement by iC3b-coated targets. Our findings demonstrate that the reduction in CR3-mediated phagocytosis associated with the 77H CD11b variant is not macrophage-restricted but demonstrable in other CR3-expressing professional phagocytic cells. The association between 77H and susceptibility to systemic lupus erythematosus most likely relates to impaired waste disposal, a key component of lupus pathogenesis.     

Urease and Serine Protease inhibitory alkaloids from Isatis tinctoria

Phytochemical investigations on the alkaloidal fraction of the whole plant of the Isatis tinctoria led to the isolation of the alkaloids 1-6., 3′-Hydroxyepiglucoisatisin (3), Epiglucoisatisin (2) were found to be potent urease inhibitors in a concentration-dependent manner with IC₅₀ values 25.63 ± 0.74, 37.01 ± 0.41 and 31.72 ± 0.93, 47.33 ± 0.31 μM against Bacillus pasteurii & Jack bean urease, respectively. Compounds 3 and 2 also showed potent inhibitory potential against α-chymotrypsin with IC₅₀ values of 23.40 ± 0.21 and 27.45 ± 0.23 μM, respectively.     

Focus Issue on Calcium Signaling: Calcium-Dependent and -Independent Stomatal Signaling Network and Compensatory Feedback Control of Stomatal Opening via Ca2+ Sensitivity Priming

Guard cells use compensatory feedback controls to adapt to conditions that produce excessively open stomata.In the past 15 years or more, many mutants that are impaired in stimulus-induced stomatal closing and opening have been identified and functionally characterized in Arabidopsis (Arabidopsis thaliana), leading to a mechanistic understanding of the guard cell signal transduction network. However, evidence has only recently emerged that mutations impairing stomatal closure, in particular those in slow anion channel SLOW ANION CHANNEL-ASSOCIATED1 (SLAC1), unexpectedly also exhibit slowed stomatal opening responses. Results suggest that this compensatory slowing of stomatal opening can be attributed to a calcium-dependent posttranslational down-regulation of stomatal opening mechanisms, including down-regulation of inward K⁺ channel activity. Here, we discuss this newly emerging stomatal compensatory feedback control model mediated via constitutive enhancement (priming) of intracellular Ca²⁺ sensitivity of ion channel activity. The CALCIUM-DEPENDENT PROTEIN KINASE6 (CPK6) is strongly activated by physiological Ca²⁺ elevations and a model is discussed and open questions are raised for cross talk among Ca²⁺-dependent and Ca²⁺-independent guard cell signal transduction pathways and Ca²⁺ sensitivity priming mechanisms.Stomatal pores formed by two guard cells enable CO₂ uptake from the atmosphere, but also ensure leaf cooling and provide a pulling force for nutrient uptake from the soil via transpiration. These vitally important processes are inevitably accompanied by water loss through stomata. Stomatal opening and closure is caused by the uptake and release of osmotically active substances and is tightly regulated by signaling pathways that lead to the activation or inactivation of guard cell ion channels and pumps. Potassium ions enter guard cells through the inward-rectifying K⁺ channels (K+_in) during stomatal opening and are released via outward-rectifying K⁺ channels during stomatal closure (Schroeder et al., 1987; Hosy et al., 2003; Roelfsema and Hedrich 2005). Cytosolic Ca²⁺, an important second messenger in plants, mediates ion channel regulation, particularly down-regulation of inward-conducting K+_in channels and activation of S-type anion channels, thus mediating stomatal closure and inhibiting stomatal opening (Schroeder and Hagiwara, 1989; Dodd et al., 2010; Kim et al., 2010). Stomatal closure is initiated by anion efflux via the slow S-type anion channel SLAC1 (Negi et al., 2008; Vahisalu et al., 2008; Kollist et al., 2011) and the voltage-dependent rapid R-type anion channel QUICK-ACTIVATING ANION CHANNEL1 (Meyer et al. 2010; Sasaki et al., 2010).In recent years, advances have been made toward understanding mechanisms mediating abscisic acid (ABA)-induced stomatal closure (Cutler et al., 2010; Kim et al., 2010; Raghavendra et al., 2010). The core ABA signaling module, consisting of PYR/RCAR (for pyrabactin resistance 1/regulatory components of ABA receptors) receptors, clade A protein phosphatases (PP2Cs), SNF-related protein kinase OPEN STOMATA1 (OST1), and downstream targets, is Ca²⁺-independent (Ma et al., 2009; Park et al., 2009; Hubbard et al., 2010). However, ABA-induced stomatal closure was reduced to only 30% of the normal stomatal closure response under conditions that inhibited intracellular cytosolic free calcium ([Ca2+]_cyt) elevations in Arabidopsis (Siegel et al., 2009), consistent with previous findings in other plants (De Silva et al., 1985; Schwartz, 1985; McAinsh et al., 1991; MacRobbie, 2000). Together these and other studies show the importance of [Ca2+]_cyt for a robust ABA-induced stomatal closure. Here, we discuss Ca²⁺-dependent and Ca²⁺-independent signaling pathways in guard cells and open questions on how these may work together.Plants carrying mutations in the SLAC1 anion channel have innately more open stomata, and exhibit clear impairments in ABA-, elevated CO₂-, Ca²⁺-, ozone-, air humidity-, darkness-, and hydrogen peroxide-induced stomatal closure (Negi et al., 2008; Vahisalu et al., 2008; Merilo et al., 2013). Recent research, however, unexpectedly revealed that mutations in SLAC1 also down-regulate stomatal opening mechanisms and slow down stomatal opening (Laanemets et al., 2013).