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Bibhuti Ranjan, Santhosh Pillai, Kugenthiren Permaul, Suren Singh
A recombinant Pichia pastoris harbouring the cyanate hydratase gene (rTl-Cyn) from the thermophilic fungus Thermomyces lanuginosus SSBP yielded a high titre of extracellular cyanate hydratase (100±13UmL(-1)) which was ∼10-fold higher than the native fungal strain. The purified rTl-Cyn had a molecular mass of ∼20kDa on SDS-PAGE, with Km, Vmax, kcat and kcat/Km values of 0.34mM, 2857.14µmolesmg(-1)min(-1), 2.14×10(4)s(-1) and 6.3 ×10(7)M(-1)s(-1), respectively. Its properties of thermostability, pH stability, and heavy metals insensitivity, make it a suitable candidate for bioremediation in extreme environments...
August 2017: Bioresource Technology
Michael Jensen, Ronald Davis
Methods of error filtration and correction post-gene assembly are a major bottleneck in the synthetic biology pipeline. Current oligonucleotide purification strategies, including polyacrylamide gel electrophoresis and high-performance liquid chromatography, are often expensive and labor-intensive, give low mass recovery, and contain hazardous chemicals. To circumvent these limitations, we explored an enzymatic means of oligonucleotide purification using RecJ, which is the only known exonuclease to digest single-stranded DNA (ssDNA) in the 5' to 3' direction...
May 9, 2017: Biochemistry
Melvin Makolomakwa, Adarsh Kumar Puri, Kugen Permaul, Suren Singh
Phytase production by the thermophilic mould Thermomyces lanuginosus SSBP was enhanced 8.56-fold in submerged fermentation, which was further improved in fed-batch cultivations. The protein was purified to homogeneity using ammonium sulphate precipitation, Resource Q anion exchange and Superdex gel-filtration chromatography, with an overall purification of 24.7-fold and a yield of 5.16%. The purified 49kDa protein was optimally active at 55°C and pH 5.0, and was stable between 50 and 90°C from pH 3.0-6.0, with a half-life of 138...
July 2017: Bioresource Technology
Antonin Bourdieu, Maryse Avalon, Véronique Lapostolle, Sadek Ismail, Margaux Mombled, Christelle Debeissat, Marianne Guérinet, Pascale Duchez, Jean Chevaleyre, Marija Vlaski-Lafarge, Arnaud Villacreces, Vincent Praloran, Zoran Ivanovic, Philippe Brunet de la Grange
Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood...
March 1, 2017: Journal of Cellular Physiology
M Föcking, P Dicker, L M Lopez, M Hryniewiecka, K Wynne, J A English, G Cagney, D R Cotter
The postsynaptic density (PSD) contains a complex set of proteins of known relevance to neuropsychiatric disorders such as schizophrenia and bipolar disorder. We enriched for this anatomical structure in the anterior cingulate cortex of 16 bipolar disorder samples and 20 controls from the Stanley Medical Research Institute. Unbiased shotgun proteomics incorporating label-free quantitation was used to identify differentially expressed proteins. Quantitative investigation of the PSD identified 2033 proteins, among which 288 were found to be differentially expressed...
November 29, 2016: Translational Psychiatry
Damiano Rizzoni
OBJECTIVE: The dose-effect relationship of fixed-dose combinations of anti-hypertensive drugs has been only poorly explored. This pooled analysis investigates the dose-response relationship of fixed-dose lercanidipine + enalapril in patients with mild-to-moderate hypertension. RESEARCH DESIGN AND METHODS: This was an individual patient data analysis of four randomized studies (n = 2340). MAIN OUTCOME MEASURES: The primary efficacy variable was the change from baseline in sitting diastolic blood pressure (SDBP)...
October 2016: Current Medical Research and Opinion
Zheng Shi, Rong Sun, Tian Yu, Rong Liu, Li-Jia Cheng, Jin-Ku Bao, Liang Zou, Yong Tang
Plant lectins have been investigated to elucidate their complicated mechanisms due to their remarkable anticancer activities. Although plant lectins seems promising as a potential anticancer agent for further preclinical and clinical uses, further research is still urgently needed and should include more focus on molecular mechanisms. Herein, a Naïve Bayesian model was developed to predict the protein-protein interaction (PPI), and thus construct the global human PPI network. Moreover, multiple sources of biological data, such as smallest shared biological process (SSBP), domain-domain interaction (DDI), gene co-expression profiles and cross-species interolog mapping were integrated to build the core apoptotic PPI network...
February 8, 2016: International Journal of Molecular Sciences
Vivek Kumar Prasad, Clemens Drenowatz, Gregory A Hand, Carl J Lavie, Xuemei Sui, Madison Demello, Steven N Blair
OBJECTIVES: We examined the independent and combined associations of cardiorespiratory fitness (CRF), body fat (BF) percentage (BF%), and body mass index (BMI) with submaximal systolic blood pressure (SSBP) among young adult women. MATERIALS AND METHODS: Analyses included a sample of 211 normotensive women with a BMI between 20 and 35 kg/m(2); BF% was calculated using total BF measured from dual X-ray absorptiometry, CRF was assessed using a graded exercise test, and SSBP was measured at each stage...
September 2016: Journal of Women's Health
Fiona Houston, Wilfred Goldmann, James Foster, Lorenzo González, Martin Jeffrey, Nora Hunter
Sheep are natural hosts of the prion disease, scrapie. They are also susceptible to experimental challenge with various scrapie strains and with bovine spongiform encephalopathy (BSE), which affects cattle and has been accidentally transmitted to a range of other species, including man. Incidence and incubation period of clinical disease in sheep following inoculation is controlled by the PRNP gene, which has different alleles defined on the basis of polymorphisms, particularly at codons 136, 154 and 171, although other codons are associated with survival time, and the exact responses of the sheep may be influenced by other breed-related differences...
2015: PloS One
Anton G Gossner, John Hopkins
Accumulation of the misfolded prion protein, PrP(Sc) in the central nervous system (CNS) is strongly linked to progressive neurodegenerative disease. For many transmissible spongiform encephalopathies (TSEs), peripheral lymphoid tissue is an important site of PrP(Sc) amplification but without gross immunological consequence. Susceptible VRQ homozygous New Zealand Cheviot sheep were infected with SSBP/1 scrapie by inoculation in the drainage area of the prescapular lymph nodes. The earliest time that PrP(Sc) was consistently detected by immunohistology in these nodes was D50 post infection...
December 31, 2015: Veterinary Microbiology
Faez Iqbal Khan, Krishna Bisetty, Suren Singh, Kugen Permaul, Md Imtaiyaz Hassan
Chitinases are ubiquitous class of extracellular enzymes, which have gained attention in the past few years due to their wide biotechnological applications. The effectiveness of conventional insecticides is increasingly compromised by the occurrence of resistance; thus, chitinase offers a potential alternative to the use of chemical fungicides. The thermostable enzymes from thermophilic microorganisms have numerous industrial, medical, environmental and biotechnological applications due to their high stability for temperature and pH...
November 2015: Extremophiles: Life Under Extreme Conditions
Vivek K Prasad, Clemens Drenowatz, Gregory A Hand, Carl J Lavie, Xuemei Sui, Madison Demello, Steven N Blair
OBJECTIVES: Exercise blood pressure is a marker of future cardiovascular morbidity and mortality among individuals with or without high resting BP or any other cardiovascular disease (CVD) signs and symptoms at present. The purpose of this study was to evaluate the association between SBP during submaximal exercise and cardiorespiratory fitness (CRF) among young men. Further, we investigated the trend displayed by submaximal SBP (SSBP) across different levels of CRF. METHODS: Graded exercise test (GXT) using a Modified Bruce protocol was performed on 204 men; SSBP was recorded at each stage of the protocol...
November 2015: Journal of Hypertension
Vashni Gramany, Faez Iqbal Khan, Algasan Govender, Krishna Bisetty, Suren Singh, Kugenthiren Permaul
The aim of this study was to clone, express, and characterize a β-xylosidase (Tlxyn1) from the thermophilic fungus Thermomyces lanuginosus SSBP in Pichia pastoris GS115 as well as analyze optimal activity and stability using computational and experimental methods. The enzyme was constitutively expressed using the GAP promoter and secreted into the medium due to the alpha-mating factor secretion signal present on the expression vector pBGPI. The 1276 bp gene consists of an open reading frame that does not contain introns...
August 2016: Journal of Biomolecular Structure & Dynamics
Enric Vidal, Natalia Fernández-Borges, Belén Pintado, Hasier Eraña, Montserrat Ordóñez, Mercedes Márquez, Francesca Chianini, Dolors Fondevila, Manuel A Sánchez-Martín, Olivier Andreoletti, Mark P Dagleish, Martí Pumarola, Joaquín Castilla
Interspecies transmission of prions is a well-established phenomenon, both experimentally and under field conditions. Upon passage through new hosts, prion strains have proven their capacity to change their properties and this is a source of strain diversity which needs to be considered when assessing the potential risks associated with consumption of prion contaminated protein sources. Rabbits were considered for decades to be a prion resistant species until proven otherwise recently. To determine the extent of rabbit susceptibility to prions and to assess the effects of passage of different prion strains through this species a transgenic mouse model overexpressing rabbit PrPC was developed (TgRab)...
August 2015: PLoS Pathogens
Faez Iqbal Khan, Algasan Govender, Kugen Permaul, Suren Singh, Krishna Bisetty
Thermomyces lanuginosus is a thermophilic fungus that produces large number of industrially-significant enzymes owing to their inherent stability at high temperatures and wide range of pH optima, including thermostable chitinases that have not been fully characterized. Here, we report cloning, characterization and structure prediction of a gene encoding thermostable chitinase II. Sequence analysis revealed that chitinase II gene encodes a 343 amino acid protein of molecular weight 36.65kDa. Our study reports that chitinase II exhibits a well-defined TIM-barrel topology with an eight-stranded α/β domain...
June 7, 2015: Journal of Theoretical Biology
Michael E Thase, Rana Fayyad, Ru-Fong J Cheng, Christine J Guico-Pabia, Jonathan Sporn, Matthieu Boucher, Karen A Tourian
OBJECTIVE: To evaluate the effect of the serotonin-norepinephrine re-uptake inhibitor desvenlafaxine on blood pressure and incidence of new onset hypertension in pooled short-term studies and in two longer-term, randomized withdrawal studies. RESEARCH DESIGN AND METHODS: Data from patients randomly assigned to desvenlafaxine 10 mg to 400 mg/day or placebo in 11 short-term (8-12 weeks), fixed-dose, double-blind, placebo-controlled studies of major depressive disorder (MDD) were pooled for analysis; two desvenlafaxine randomized withdrawal studies (36 and 46 weeks) were analyzed separately...
April 2015: Current Medical Research and Opinion
A M Winger, J L Heazlewood, L J G Chan, C J Petzold, K Permaul, S Singh
Thermomyces lanuginosus is a thermophilic fungus known for its ability to produce industrially important enzymes including large amounts of xylanase, the key enzyme in hemicellulose hydrolysis. The secretome of T. lanuginosus SSBP was profiled by shotgun proteomics to elucidate important enzymes involved in hemicellulose saccharification and to characterise the presence of other industrially interesting enzymes. This study reproducibly identified a total of 74 proteins in the supernatant following growth on corn cobs...
November 2014: Journal of Industrial Microbiology & Biotechnology
Seung Hun Jeong, Hyoung Kyu Kim, In-Sung Song, Su Jin Noh, Jubert Marquez, Kyung Soo Ko, Byoung Doo Rhee, Nari Kim, Natalia P Mishchenko, Sergey A Fedoreyev, Valentin A Stonik, Jin Han
Echinochrome A (Ech A) is a natural pigment from sea urchins that has been reported to have antioxidant properties and a cardio protective effect against ischemia reperfusion injury. In this study, we ascertained whether Ech A enhances the mitochondrial biogenesis and oxidative phosphorylation in rat cardio myoblast H9c2 cells. To study the effects of Ech A on mitochondrial biogenesis, we measured mitochondrial mass, level of oxidative phosphorylation, and mitochondrial biogenesis regulatory gene expression...
August 2014: Marine Drugs
Anton G Gossner, John Hopkins
Sheep scrapie is a transmissible spongiform encephalopathy (TSE), progressive and fatal neurodegenerative diseases of the central nervous system (CNS) linked to the accumulation of misfolded prion protein, PrP(Sc). New Zealand Cheviot sheep, homozygous for the VRQ genotype of the PRNP gene are most susceptible with an incubation period of 193 days with SSBP/1 scrapie. However, the earliest time point that PrP(Sc) can be detected in the CNS is 125 days (D125). The aim of this study was to quantify changes to the transcriptome of the thalamus and obex (medulla) at times immediately before (D75) and after (D125) PrP(Sc) was detected...
October 10, 2014: Veterinary Microbiology
Shunsuke Takahashi, Tomohiro Usui, Shohei Kawasaki, Hidefumi Miyata, Hirofumi Kurita, Shun-ichi Matsuura, Akira Mizuno, Masahiko Oshige, Shinji Katsura
T7 Exonuclease (T7 Exo) DNA digestion reactions were studied using direct single-molecule observations in microflow channels. DNA digestion reactions were directly observed by staining template DNA double-stranded regions with SYTOX Orange and staining single-stranded (digested) regions with a fluorescently labeled ssDNA-recognizing peptide (ssBP-488). Sequentially acquired photographs demonstrated that a double-stranded region monotonously shortened as a single-stranded region monotonously increased from the free end during a DNA digestion reaction...
July 15, 2014: Analytical Biochemistry
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