Development of mixed inoculum for methane enriched biogas production

Inocula were collected from four different sources such as Jajmau tannery waste treatment plant (ITW), Jajmau municipal waste treatment (IMW), Unnao distillery (IDW) and a batch reactor, in which the sludge of a field scale biogas reactor was added to cow dung slurry to develop inoculum (IBS). A combination of these mixed inocula were used for biogas production at 35°C in laboratory scale reactor (10 L capacity) and the average yield of biogas (0.547 Lg^?1 volatile solid (VS)) and methane (0.323 Lg^?1VS) in 41 d was higher in case of mixed inoculum IMW ₁ (IMW+IBS), with maximum methane content in biogas (68% during 27–30 d), as compared to other mixed inocula as well as control i.e. ITW ₁ (ITW+IBS), IDW₁ (IDW+IBS) and IBS. The corresponding yields of gas were biogas (0.505, 0.536 and 0.456 Lg^?1VS), methane (0.288, 0.305, and 0.245 Lg^?1VS) where as, the corresponding maximum methane content in biogas was 62% during 29–33d, 64% during 29–33 d and 62% during 27–29 d in ITW₁, IDW₁ and IBS.     

Induction of male and female sterility in isabgol (<Emphasis Type="Italic">Plantago ovata</Emphasis>) due to floral infection of downy mildew (<Emphasis Type="Italic">Pernospora plantaginis</Emphasis>)

Downy mildew (Peronospora plantaginis) caused two different types of infection in the floral parts of isabgol (Plantago ovata). Systemic infection resulted in long spikes bearing weak and sterile florets, which later turned black due to saprophytic growth. Localised infection produced various symptoms ranging between normal flower opening and failure to bloom. Different parts of infected flowers such as sepal, petal, filament and anther were reduced in size compared to healthy flowers. However, gynoecium was elongated in localised infection. P. plantaginis induced gradual sterility of isabgol flowers. Androecium was affected more than the gynoecium was. Pollen number, pollen viability and germination reduced drastically due to localised infection. On the contrary, there were no significant differences between healthy and locally infected flowers in terms of stigma receptivity. In systemically infected spikes, bud development was arrested leading to sterility. When localised disease severity was high, secondary systemic infection caused similar symptoms. Microscopic observations showed presence of the pathogen in different parts of the flowers. Downy mildew adversely affected seed yield and quality; producing seeds, which were smaller and lighter than the healthy ones and later, became black. Seed yield was reduced by as much as 73.45 percent. Husk content per unit seed mass increased relatively as the total surface area of infected seeds increased.     

CABYR isoforms expressed in late steps of spermiogenesis bind with AKAPs and ropporin in mouse sperm fibrous sheath

Background  

CABYR is a polymorphic calcium-binding protein of the sperm fibrous sheath (FS) which gene contains two coding regions (CR-A and CR-B) and is tyrosine as well as serine/threonine phosphorylated during in vitro sperm capacitation. Thus far, the detailed information on CABYR protein expression in mouse spermatogenesis is lacking. Moreover, because of the complexity of this polymorphic protein, there are no data on how CABYR isoforms associate and assemble into the FS.     

Proteomics Characterization of Extracellular Space Components in the Human Aorta

The vascular extracellular matrix (ECM) is essential for the structural integrity of the vessel wall and also serves as a substrate for the binding and retention of secreted products of vascular cells as well as molecules coming from the circulation. Although proteomics has been previously applied to vascular tissues, few studies have specifically targeted the vascular ECM and its associated proteins. Thus, its detailed composition remains to be characterized. In this study, we describe a methodology for the extraction of extracellular proteins from human aortas and their identification by proteomics. The approach is based on (a) effective decellularization to enrich for scarce extracellular proteins, (b) successful solubilization and deglycosylation of ECM proteins, and (c) relative estimation of protein abundance using spectral counting. Our three-step extraction approach resulted in the identification of 103 extracellular proteins of which one-third have never been reported in the proteomics literature of vascular tissues. In particular, three glycoproteins (podocan, sclerostin, and agrin) were identified for the first time in human aortas at the protein level. We also identified extracellular adipocyte enhancer-binding protein 1, the cartilage glycoprotein asporin, and a previously hypothetical protein, retinal pigment epithelium (RPE) spondin. Moreover, our methodology allowed us to screen for proteolysis in the aortic samples based on the identification of proteolytic enzymes and their corresponding degradation products. For instance, we were able to detect matrix metalloproteinase-9 by mass spectrometry and relate its presence to degradation of fibronectin in a clinical specimen. We expect this proteomics methodology to further our understanding of the composition of the vascular extracellular environment, shed light on ECM remodeling and degradation, and provide insights into important pathological processes, such as plaque rupture, aneurysm formation, and restenosis.Vascular cells, in particular vascular smooth muscle cells, produce and maintain a complex meshwork of ECM.¹ The ECM is not only the scaffold for the anchorage and mobility of residing cells but also absorbs and transduces the shear and strain forces of the blood flow. It is primarily composed of elastin, collagen, proteoglycans, and glycoproteins. The elastin fibers and type I and III fibrillar collagens form a rigid network of highly cross-linked interstitial matrix. They offer elasticity (elastin) and tensile strength (collagens). Proteoglycans, because of their negative charge, attract water and confer resistance to compression. Finally, glycoproteins participate in matrix organization and are essential for cell attachment.The vascular ECM also serves as a substrate for the binding and retention of secreted, soluble proteins of vascular cells as well as molecules coming from the circulation, including lipoproteins, growth factors, cytokines, proteases, and protease inhibitors. These components are invariably associated with ECM proteins, especially proteoglycans. Together they comprise the vascular extracellular environment and are pivotal for disease processes, such as atherosclerosis and aneurysm formation (1).Although proteomics has been previously applied to vascular tissues, only one study has specifically targeted the extracellular vascular environment (2). This study was focused on the isolation of intimal proteoglycans from human carotid arteries. Moreover, most proteomics studies use whole tissue lysates, which are rich in cellular proteins that inevitably mask the identification of the less abundant proteins of the vascular extracellular environment (3–5). Thus, the composition of the vascular ECM and its associated proteins remains poorly defined. In the present study, we used morphologically normal human aortic samples to develop a method for the extraction of proteins present in the extracellular environment, including ECM proteins and proteins attached to the ECM. We had three specific aims: first, to reduce the contamination with cellular proteins, thereby increasing the chance of identifying scarce extracellular proteins; second, to efficiently solubilize and deglycosylate ECM proteins to improve their analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS); and third, to interface the nanoflow LC system to a recently developed injection device, which splits the flow from the analytical column, to allow the reanalysis of the same sample during a single LC-MS/MS run (RePlay, Advion).Our methodology provides a detailed overview of the aortic ECM and its associated proteins, many reported for the first time in proteomics analysis of the vasculature. Most importantly, this method could be adapted for use with other tissues to further our understanding of the composition of extracellular environment and ECM turnover under various disease conditions.     

“Working the System”—British American Tobacco's Influence on the European Union Treaty and Its Implications for Policy: An Analysis of Internal Tobacco Industry Documents

Background

Impact assessment (IA) of all major European Union (EU) policies is now mandatory. The form of IA used has been criticised for favouring corporate interests by overemphasising economic impacts and failing to adequately assess health impacts. Our study sought to assess how, why, and in what ways corporations, and particularly the tobacco industry, influenced the EU''s approach to IA.

Methods and Findings

In order to identify whether industry played a role in promoting this system of IA within the EU, we analysed internal documents from British American Tobacco (BAT) that were disclosed following a series of litigation cases in the United States. We combined this analysis with one of related literature and interviews with key informants. Our analysis demonstrates that from 1995 onwards BAT actively worked with other corporate actors to successfully promote a business-oriented form of IA that favoured large corporations. It appears that BAT favoured this form of IA because it could advance the company''s European interests by establishing ground rules for policymaking that would: (i) provide an economic framework for evaluating all policy decisions, implicitly prioritising costs to businesses; (ii) secure early corporate involvement in policy discussions; (iii) bestow the corporate sector with a long-term advantage over other actors by increasing policymakers'' dependence on information they supplied; and (iv) provide businesses with a persuasive means of challenging potential and existing legislation. The data reveal that an ensuing lobbying campaign, largely driven by BAT, helped secure binding changes to the EU Treaty via the Treaty of Amsterdam that required EU policymakers to minimise legislative burdens on businesses. Efforts subsequently focused on ensuring that these Treaty changes were translated into the application of a business orientated form of IA (cost–benefit analysis [CBA]) within EU policymaking procedures. Both the tobacco and chemical industries have since employed IA in apparent attempts to undermine key aspects of European policies designed to protect public health.

Conclusions

Our findings suggest that BAT and its corporate allies have fundamentally altered the way in which all EU policy is made by making a business-oriented form of IA mandatory. This increases the likelihood that the EU will produce policies that advance the interests of major corporations, including those that produce products damaging to health, rather than in the interests of its citizens. Given that the public health community, focusing on health IA, has largely welcomed the increasing policy interest in IA, this suggests that urgent consideration is required of the ways in which IA can be employed to undermine, as well as support, effective public health policies. Please see later in the article for the Editors'' Summary     

Putative Location of Common Region and Coat Protein Gene of Blackgram Isolate of Mungbean Yellow Mosaic Geminivirus

Reciprocal cross hybridization between DNA-A and ONA-B molecules showed that MYMV genome contains certain regions common to each other. “Common region” of MYMV belongs to the 0.55 kb Kpnl-Clal fragment of DNA-A and 0.25 kb Xbal-Hind III fragment of DNA-B. Hybridization of DNA-A restriction fragments with the full length coat protein region probe to Indian cassava mosaic virus (ICMV) showed the location of MYMV coat protein coding region in DNA-A. The greater extent of homology of ICMV coat protein region falls within the 400 base pair region of Clal-Clal fragment in DNA-A of MYMV-Bg. The homology between ICMV and MYMV-Bg coat protein gene appeared to be less than 50%. Cross hybridization between DNA-A and DNA-B molecules helped to trim out the homologous region and the DNA-A and DNA-B specific probes could be tailored.     

The chromogranin A peptide vasostatin-I inhibits gap formation and signal transduction mediated by inflammatory agents in cultured bovine pulmonary and coronary arterial endothelial cells

The proinflammatory agent tumour necrosis factor alpha (TNFalpha) is one of several agents causing vascular leakage. The N-terminal domain of CgA, vasostatin-I (CgA1-76), has recently been reported to inhibit TNFalpha induced gap formation in human umbilical venous endothelial cells. Here we report on the effect of recombinant human CgA1-78, vasostatin-I, on TNFalpha induced gap formation in two model systems of vascular leakage in arterial endothelial cells of bovine pulmonary (BPAEC) and coronary (BCAEC) origin. Vasostatin-I inhibited the TNFalpha induced gap formation in both models, being inactive in the unstimulated cells. The phosphorylation of p38MAP kinase in TNFalpha activated BPAEC was markedly attenuated in the presence of vasostatin-I and the inhibitory effect corresponded to that of the specific p38MAPK inhibitor SB203580. Vasostatin-I also inhibited the phosphorylation of p38MAPK induced by both thrombin and pertussis toxin in these cells. The results demonstrate that vasostatin-I has inhibitory effects on TNFalpha-induced disruption of confluent layers of cultured pulmonary and coronary arterial endothelial cells. This suggests that vasostatin-I may affect endothelial barrier dysfunction also in arterial vascular beds. Furthermore, the inhibitory activity of vasostatin-I may be associated with the p38MAPK signalling cascade via a pertussis toxin sensitive, presumably Galphai coupled mechanism.     

Alginate encapsulation of axillary buds of Ocimum americanum L. (hoary basil), O. Basilicum L. (sweet basil), O. Gratissimum L. (shrubby basil), and O. Sanctum. L. (sacred basil)

Summary Propagation and conservation of four pharmaceutically important herbs, Ocimum americanum L. syn. O. canum Sims. (hoary basil); O basilicum L. (swett basil); O. gratissimum L. (shrubby basil); and O. sanctum L. (sacred basil) was attempted using synthetic seed technology. Synthetic seeds were produced by encapsulating axillary vegetative buds harvested from garden-grown plants of these four Ocimum species in calcium alginate gel. The gel contained Murashige and Skoog (MS) nutrients and 1.1-4.4 μM benzyladenine (BA). Shoots emerged from the encapsulated buds on all six planting media tested. However, the highest frequency shoot emergence and maximum number of shoots per bud were recorded on media containing BA. Of the six planting media tested, both shoot and root emergence from the encapsulated buds in a single step was recorded on growth regulator-free MS medium as well as on vermi-compost moistened with halfstrength MS medium. Rooted shoots were retrieved from the encapsulated buds of O. americanum, O. basilicum, and O. sanctum on these two media, whereas shoots of O. gratissimum failed to root. The encapsulated buds could be stored for 60 d at 4°C. Plants retrieved from the encapsulated buds were hardened off and established in soil.