Actinobacillus pleuropneumoniae (App) is the etiological agent of acute porcine pneumonia and responsible for severe economic losses worldwide. The capsule polymer of App serotype 1 (App1) consists of [4)-GlcNAc-β(1,6)-Gal-α-1-(PO4-] repeating units that are O-acetylated at O-6 of the GlcNAc. It is a major virulence factor and was used in previous studies in the successful generation of an experimental glycoconjugate vaccine. However, the application of glycoconjugate vaccines in the animal health sector is limited, presumably because of the high costs associated with harvesting the polymer from pathogen culture. Consequently, here we exploited the capsule polymerase Cps1B of App1 as an in vitro synthesis tool and an alternative for capsule polymer provision. Cps1B consists of two catalytic domains, as well as a domain rich in tetratricopeptide repeats (TPRs). We compared the elongation mechanism of Cps1B with that of a ΔTPR truncation (Cps1B-ΔTPR). Interestingly, the product profiles displayed by Cps1B suggested processive elongation of the nascent polymer, whereas Cps1B-ΔTPR appeared to work in a more distributive manner. The dispersity of the synthesized products could be reduced by generating single-action transferases and immobilizing them on individual columns, separating the two catalytic activities. Furthermore, we identified the O-acetyltransferase Cps1D of App1 and used it to modify the polymers produced by Cps1B. Two-dimensional NMR analyses of the products revealed O-acetylation levels identical to those of polymer harvested from App1 culture supernatants. In conclusion, we have established a protocol for the pathogen-free in vitro synthesis of tailored, nature-identical App1 capsule polymers.

Lens proteins become increasingly cross-linked through nondisulfide linkages during aging and cataract formation. One mechanism that has been implicated in this cross-linking is glycation through formation of advanced glycation end products (AGEs). Here, we found an age-associated increase in stiffness in human lenses that was directly correlated with levels of protein–cross-linking AGEs. α-Crystallin in the lens binds to other proteins and prevents their denaturation and aggregation through its chaperone-like activity. Using a FRET-based assay, we examined the stability of the αA-crystallin–γD-crystallin complex for up to 12 days and observed that this complex is stable in PBS and upon incubation with human lens–epithelial cell lysate or lens homogenate. Addition of 2 mm ATP to the lysate or homogenate did not decrease the stability of the complex. We also generated complexes of human αA-crystallin or αB-crystallin with alcohol dehydrogenase or citrate synthase by applying thermal stress. Upon glycation under physiological conditions, the chaperone–client complexes underwent greater extents of cross-linking than did uncomplexed protein mixtures. LC-MS/MS analyses revealed that the levels of cross-linking AGEs were significantly higher in the glycated chaperone–client complexes than in glycated but uncomplexed protein mixtures. Mouse lenses subjected to thermal stress followed by glycation lost resilience more extensively than lenses subjected to thermal stress or glycation alone, and this loss was accompanied by higher protein cross-linking and higher cross-linking AGE levels. These results uncover a protein cross-linking mechanism in the lens and suggest that AGE-mediated cross-linking of α-crystallin–client complexes could contribute to lens aging and presbyopia.

The dextransucrase DSR-OK from the Gram-positive bacterium Oenococcus kitaharae DSM17330 produces a dextran of the highest molar mass reported to date (∼109 g/mol). In this study, we selected a recombinant form, DSR-OKΔ1, to identify molecular determinants involved in the sugar polymerization mechanism and that confer its ability to produce a very-high-molar-mass polymer. In domain V of DSR-OK, we identified seven putative sugar-binding pockets characteristic of glycoside hydrolase 70 (GH70) glucansucrases that are known to be involved in glucan binding. We investigated their role in polymer synthesis through several approaches, including monitoring of dextran synthesis, affinity assays, sugar binding pocket deletions, site-directed mutagenesis, and construction of chimeric enzymes. Substitution of only two stacking aromatic residues in two consecutive sugar-binding pockets (variant DSR-OKΔ1-Y1162A-F1228A) induced quasi-complete loss of very-high-molar-mass dextran synthesis, resulting in production of only 10–13 kg/mol polymers. Moreover, the double mutation completely switched the semiprocessive mode of DSR-OKΔ1 toward a distributive one, highlighting the strong influence of these pockets on enzyme processivity. Finally, the position of each pocket relative to the active site also appeared to be important for polymer elongation. We propose that sugar-binding pockets spatially closer to the catalytic domain play a major role in the control of processivity. A deep structural characterization, if possible with large-molar-mass sugar ligands, would allow confirming this hypothesis.

Bacterial biofilms are cellular communities that produce an adherent matrix. Exopolysaccharides are key structural components of this matrix and are required for the assembly and architecture of biofilms produced by a wide variety of microorganisms. The human bacterial pathogens Escherichia coli and Salmonella enterica produce a biofilm matrix composed primarily of the exopolysaccharide phosphoethanolamine (pEtN) cellulose. Once thought to be composed of only underivatized cellulose, the pEtN modification present in these matrices has been implicated in the overall architecture and integrity of the biofilm. However, an understanding of the mechanism underlying pEtN derivatization of the cellulose exopolysaccharide remains elusive. The bacterial cellulose synthase subunit G (BcsG) is a predicted inner membrane–localized metalloenzyme that has been proposed to catalyze the transfer of the pEtN group from membrane phospholipids to cellulose. Here we present evidence that the C-terminal domain of BcsG from E. coli (EcBcsGΔN) functions as a phosphoethanolamine transferase in vitro with substrate preference for cellulosic materials. Structural characterization of EcBcsGΔN revealed that it belongs to the alkaline phosphatase superfamily, contains a Zn2+ ion at its active center, and is structurally similar to characterized enzymes that confer colistin resistance in Gram-negative bacteria. Informed by our structural studies, we present a functional complementation experiment in E. coli AR3110, indicating that the activity of the BcsG C-terminal domain is essential for integrity of the pellicular biofilm. Furthermore, our results established a similar but distinct active-site architecture and catalytic mechanism shared between BcsG and the colistin resistance enzymes.

Extracellular matrix-evoked angiostasis and autophagy within the tumor microenvironment represent two critical, but unconnected, functions of the small leucine-rich proteoglycan, decorin. Acting as a partial agonist of vascular endothelial growth factor 2 (VEGFR2), soluble decorin signals via the energy sensing protein, AMP-activated protein kinase (AMPK), in the autophagic degradation of intracellular vascular endothelial growth factor A (VEGFA). Here, we discovered that soluble decorin evokes intracellular catabolism of endothelial VEGFA that is mechanistically independent of mTOR, but requires an autophagic regulator, paternally expressed gene 3 (PEG3). We found that administration of autophagic inhibitors such as chloroquine or bafilomycin A1, or depletion of autophagy-related 5 (ATG5), results in accumulation of intracellular VEGFA, indicating that VEGFA is a basal autophagic substrate. Mechanistically, decorin increased the VEGFA clearance rate by augmenting autophagic flux, a process that required RAB24 member RAS oncogene family (RAB24), a small GTPase that facilitates the disposal of autophagic compartments. We validated these findings by demonstrating the physiological relevance of this process in vivo. Mice starved for 48 h exhibited a sharp decrease in overall cardiac and aortic VEGFA that could be blocked by systemic chloroquine treatment. Thus, our findings reveal a unified mechanism for the metabolic control of endothelial VEGFA for autophagic clearance in response to decorin and canonical pro-autophagic stimuli. We posit that the VEGFR2/AMPK/PEG3 axis integrates the anti-angiogenic and pro-autophagic bioactivities of decorin as the molecular basis for tumorigenic suppression. These results support future therapeutic use of decorin as a next-generation protein therapy to combat cancer.

Multidrug resistance (MDR) in cancer arises from cross-resistance to structurally- and functionally-divergent chemotherapeutic drugs. In particular, MDR is characterized by increased expression and activity of ATP-binding cassette (ABC) superfamily transporters. Sphingolipids are substrates of ABC proteins in cell signaling, membrane biosynthesis, and inflammation, for example, and their products can favor cancer progression. Glucosylceramide (GlcCer) is a ubiquitous glycosphingolipid (GSL) generated by glucosylceramide synthase, a key regulatory enzyme encoded by the UDP-glucose ceramide glucosyltransferase (UGCG) gene. Stressed cells increase de novo biosynthesis of ceramides, which return to sub-toxic levels after UGCG mediates incorporation into GlcCer. Given that cancer cells seem to mobilize UGCG and have increased GSL content for ceramide clearance, which ultimately contributes to chemotherapy failure, here we investigated how inhibition of GSL biosynthesis affects the MDR phenotype of chronic myeloid leukemias. We found that MDR is associated with higher UGCG expression and with a complex GSL profile. UGCG inhibition with the ceramide analog d-threo-1-(3,4,-ethylenedioxy)phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol (EtDO-P4) greatly reduced GSL and monosialotetrahexosylganglioside levels, and co-treatment with standard chemotherapeutics sensitized cells to mitochondrial membrane potential loss and apoptosis. ABC subfamily B member 1 (ABCB1) expression was reduced, and ABCC-mediated efflux activity was modulated by competition with nonglycosylated ceramides. Consistently, inhibition of ABCC-mediated transport reduced the efflux of exogenous C6-ceramide. Overall, UGCG inhibition impaired the malignant glycophenotype of MDR leukemias, which typically overcomes drug resistance through distinct mechanisms. This work sheds light on the involvement of GSL in chemotherapy failure, and its findings suggest that targeted GSL modulation could help manage MDR leukemias.

Endorepellin, the C-terminal fragment of the heparan sulfate proteoglycan perlecan, influences various signaling pathways in endothelial cells by binding to VEGFR2. In this study, we discovered that soluble endorepellin activates the canonical stress signaling pathway consisting of PERK, eIF2α, ATF4, and GADD45α. Specifically, endorepellin evoked transient activation of VEGFR2, which, in turn, phosphorylated PERK at Thr980. Subsequently, PERK phosphorylated eIF2α at Ser51, upregulating its downstream effector proteins ATF4 and GADD45α. RNAi-mediated knockdown of PERK or eIF2α abrogated the endorepellin-mediated up-regulation of GADD45α, the ultimate effector protein of this stress signaling cascade. To functionally validate these findings, we utilized an ex vivo model of angiogenesis. Exposure of the aortic rings embedded in 3D fibrillar collagen to recombinant endorepellin for 2–4 h activated PERK and induced GADD45α vis à vis vehicle-treated counterparts. Similar effects were obtained with the established cellular stress inducer tunicamycin. Notably, chronic exposure of aortic rings to endorepellin for 7–9 days markedly suppressed vessel sprouting, an angiostatic effect that was rescued by blocking PERK kinase activity. Our findings unravel a mechanism by which an extracellular matrix protein evokes stress signaling in endothelial cells, which leads to angiostasis.

Losses within the entertainment industry are already estimated to be in the billions as the sector continues to reel from the impacts of COVID-19. Major events like Reggae Sumfest, which has already been cancelled this year, had an estimated...

New research suggests that neurons which track our movements are also involved in imaginary navigation

Brain cells involved in spatial navigation and mapping the environment also fire when we merely imagine moving through familiar surroundings, according to a new study by researchers at University College London. The research, published today in the journal Current Biology, shows that memory and imagination are intimately linked in the brain at the cellular level, and could help to explain some of the changes that occur in the early stages of Alzheimer’s Disease.

Related: The fly's neural compass works just like a mammal's

Related: 3D compass cells found in the bat brain

New research reveals the secret behind the remarkable intelligence of some bird species

Related: Blind cave fish evolved a shrunken brain to save energy

Calling someone “bird brain” used to be considered as an insult. Birds’ brains are very small compared to those of mammals, and what’s more, they lack the heavily wrinkled cerebral cortex, which is characteristic of the human brain, and widely believed to the seat of intelligence. It was, therefore, widely assumed that birds aren’t very clever creatures, but recently this has started to change.

Related: Ravens cooperate with friends not foes

New research shows that a genetic mutation associated with schizophrenia alters the process of cellular differentiation, disturbing the balance of neurons and glia in the brain

Stem cells obtained from patients with schizophrenia carry a genetic mutation that alters the ratio of the different type of nerve cells they produce, according to a new study by researchers in Japan. The findings, published today in the journal Translational Psychiatry, suggest that abnormal neural differentiation may contribute to the disease, such that fewer neurons and more non-neuronal cells are generated during the earliest stages of brain development.

Schizophrenia is a debilitating mental illness that affects about 1 in 100 people. It is known to be highly heritable, but is genetically complex: so far, researchers have identified over 100 rare genetic variations and dozens of mutations associated with increased risk of developing the disease.

Related: Brain’s immune cells hyperactive in schizophrenia

Related: Turning urine into brain cells | Mo Costandi

Epithelial cells are held together and connected by several different types of structures that form cell-cell contacts.

At CES 2020, Sony unveiled a new line of 4K and 8K TVs, destined to be released during the year. They finally have now gone on sale and we were quick to pick one up to test it out.

Shares of Idea Cellular plunged 6.99 % on Monday closing the session at Rs 85.85 a share on the Bombay Stock Exchange (BSE) after the company on Saturday reported a poor set of numbers for the March quarter.

Aggregate recast loans worth around Rs 50,000 crore (Rs 49,332 crore) failed at the corporate debt restructuring (CDR) cell in FY17, taking total failures since the inception of the cell in 2001 to Rs 1.37 lakh crore.

In another release, the ministry said the Institute of Genomics and Integrative Biology (CSIR-IGIB) and Tata Sons have signed a licensing agreement for a paper strip-based test called Feluda for Covid 19's rapid diagnosis.

The provincial government in Pakistan's Sindh province is planning a literacy program to reach women and girls in remote areas via cell phones.

PESHAWAR: Confusion prevailed in Khyber Pakhtunkhwa over cancellation of the board examinations and extension of school closure until July 15 as no decision could be taken at the provincial level despite the loud and clear announcement made by federal Education Minister Shafqat...

The body had informed Caf their concerns after the federation annulled the top tier on April 30 ......

NETANYA, Israel, May 10, 2020/PRNewswire/ -- Cellcom Israel Ltd. (NYSE: CEL) (TASE: CEL) (the 'Company') announced today that following the Company's previously announced potential debt offering in Israel, the Company is in preparations to raise up to NIS 200,000,000in debentures and options to purchase ordinary shares of the Company, and subject to the completion of the contemplated offering, the Company has accepted early commitments from institutional investors for the purchase of an aggregate of 222,000 units, each comprised of 1,000 debentures Series L and 10 options Series 4, for NIS 906per unit out of orders for approximately 560 thousand units received in the institutional tender. In...

PESHAWAR: Confusion prevailed in Khyber Pakhtunkhwa over cancellation of the board examinations and extension of school closure until July 15 as no decision could be taken at the provincial level...

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Clear Blue Technologies receives $3.5 million CAD deal to build out solar-powered cell networks in the Congo, Ghana, and Nigeria

Hi, mera gst 2sal se due hai. aur cancelled v ho gaya hai. ab kaya karu?

A long-standing mystery of how a protein displayed on the surface of malaria parasites called "TRAP" interacts with human host cells has now been found.

The Secretary-General met with Chancellor Angela Merkel and the Heads of International Organisations to discuss the global economic outlook as well as policies to boost growth and employment in Europe.

Restoring competitiveness is one of the key challenges to bring European economies back on a path of strong, sustainable and balanced growth. Europe could improve its growth prospects by implementing a strategic reform agenda with a broad range of policy reforms to increase productivity, dynamism and employment.

OECD Secretary-General Angel Gurría congratulated German Chancellor Angela Merkel on her re-election for a third term.

As part of the OECD Leaders’ Programme, Angel Gurría, Secretary-General met with Germany’s Federal Chancellor Angela Merkel to discuss competitiveness and productivity in Germany and Europe and policies to promote inclusive growth and well-being.

If the quality of an education system can never exceed the quality of its teachers, then countries need to do all they can to build a high-quality teaching force.

Tertiary qualifications have become the entrance ticket for modern societies. Never before have those with advanced qualifications had the life chances they enjoy today, and never before have those who struggled to acquire a good education paid the price they pay today.

The double-deck aircraft was rolled into scaffolding following a routine maintenance check at Sydney Airport on Friday and will now be grounded for at least two weeks.

The Australian airline cancels almost one-in-10 scheduled flights between Melbourne and Sydney in the six months to November.

The British comedian announced on Monday he decided to cancel his sell-out gig in Perth after a woman who attended the venue was found to have contracted the deadly illness.

Qantas is preparing to lose $40 million a week until the travel industry recovers from the coronavirus shutdown.

Mr, Saniay Govind Dhande & Ors. .. Respondents BERPORE:

HON BLE MR.FUSTICE SHIVA BIRTI SINGH, CHAIRPERSON For Petitioner (in C.A. No.l of 2014) =: Mr, Thyagrajan, Advocate Ms. Akanksha Banerjee, Advocate For Petitioner (in C.A. No.4 of 2014) : Mr. Alok Sharma, Advocate For Respondents > Mr. Arpun Natrajan, Advocate QRDER By S.K. Singh, Chairperson -- At the outset, it ig recorded that learned counsel for Vodafone Cellular Ltd, appellant in Cyber Appeal No.1 of 2014 has informed that the name of the above corporate entity now stands changed to Vodafone Idea Lid. He prays that this change may be recorded and the changed name should appear in the judgment. This prayer has not been opposed by the learned counsel for the other side and hence the change in the name of Vodatone Cellular Ltd. to that of Vodafone Idea Lid. is recorded and the cause title of this judgment and order is accordingly modified so as to teflect the name of Vodafone Idea Lid. Cyber Appeal No.1 of 2014 has been taken as the lead matter. The appellant in the other appeal (Cyber Appeal No.4 of 2014) Le. ICICT Bank Lid. is one of the respondents in Cyber Appeal No.1 of 2014,

Ref. Case No. 05 of 2019 1

2. The Informant in the present case had issued RFP for procurement of Cloth Cotton Pagdi for quantity of 7,42,426 Mtrs and Mattress MK-II (Improved Version), quantity 57,761 (in numbers). The Informant has stated that out of 04 firms which participated, only 03 firms could qualify for opening of commercial bids for Cloth Cotton Pagdi and out of 10 firms only 04 could qualify for opening of commercial bid for mattress. The tender for procurement of Cotton Pagdi was floated on 22.10.2018, and for Mattress was floated on 08.11.2018, respectively.

3. The Informant has averred that Commercial Negotiation Committee ('CNC') observed that the rates may have been quoted after collusion by the said two firms. As submitted by the Informant, details of the bid are reproduced in the table below:

2. The Informant in the present case had floated a Tender No. A/59919/Shirt Khakhi/DGOS/OS-PII/Proc Sec, dated 19.06.2017 for procurement of 1,38,251 Shirt Man's Cellular Cotton 1973 Pattern (Modified) Khaki ("Item"). The Informant has stated that out of 14 firms which participated, only 09 qualified for the opening of their commercial bids.

3. The Informant has averred that Commercial Negotiation Committee ('CNC') observed that the rates may have been quoted after collusion by the said four firms (Opposite Parties). As submitted by the Informant, details of the bid are reproduced in the table below:

Table 1: Details of Bidders S. No Firm Name Rate (in Rs) Status

Ref. Case No. 05 of 2019 1

2. The Informant in the present case had issued RFP for procurement of Cloth Cotton Pagdi for quantity of 7,42,426 Mtrs and Mattress MK-II (Improved Version), quantity 57,761 (in numbers). The Informant has stated that out of 04 firms which participated, only 03 firms could qualify for opening of commercial bids for Cloth Cotton Pagdi and out of 10 firms only 04 could qualify for opening of commercial bid for mattress. The tender for procurement of Cotton Pagdi was floated on 22.10.2018, and for Mattress was floated on 08.11.2018, respectively.

3. The Informant has averred that Commercial Negotiation Committee ('CNC') observed that the rates may have been quoted after collusion by the said two firms. As submitted by the Informant, details of the bid are reproduced in the table below:

2. The Informant in the present case had floated a Request for Proposal ("RFP") No. A/59876/Durries/ Clo-1/DGOS/OS-PII/Proc Sec dated 15.12.2015 for procurement of 8,18,009 Durries IT OG ("Item").

3. The Informant averred that 09 firms participated in the said tender including Standard Gram/OP-2 and out of the said 09 firms, only 06 qualified for opening of commercial bids. As stated by the Informant, Standard Gram/OP-2 could not qualify in technical evaluation as the firm was not registered with Association of Corporations and Apex Societies of Handlooms/Khadi Village Industries Commission ("ACASH/KVIC") which was a pre-requisite. It is further stated that while the contract was under progress, Standard Gram/OP-2 merged with Integrated Defence/OP-3. Subsequently, the L1 firm (HP Handicraft/OP-1) sublet the manufacture of the Item to Integrated Defence/OP-3 vide Letter No. HPSHHC:173/10(EM)/Durries/838081 dated 23.03.2018.

Rishi Sunak confirmed in today's Budget statement that taxation on petrol and diesel will remain at 57.95p per litre, as it has since March 2011.

There is cause for concern following the news that Love Island will not go ahead this summer as well as rumours circulating Strictly Come Dancing.

The quality of teaching and research in Indian universities has rapidly declined. Surely this has something to do with the manner in which we appoint those at their helm, writes Ramachandra Guha.

कोरोना वायरस संक्रमण के चलते अभी भी बहुत सी जगहों पर लॉकडाउन जारी है। लेकिन उन जगहों पर जहां कोरोना के मामले कम हैं या नहीं है, वहां पर सरकार ने व्यवसाय शुरू करने की इजाजत दी है और ऐसा हुए अभी दो ही दिन हुए हैं।

The Telangana State Council of Higher Education (TSCHE) has announced the 'Connect Chancellor' programme, which is a platform to bring out creativity, innovation and novel ideas - an opportunity to the students of state universities to write articles, stories and poems,

Singer Chinmayi Sripada has always been active on social media and never forgets to raise her voice on social issues. For the past three weeks, Chinmayi is also using her singing talent in raising funds for the families of daily wage

Archives, Room Use Only - QC761.W54 1876

The spread of the coronavirus has halted all tennis events since mid-March and will not resume until July 13 at the earliest