Leptin DNA Methylation and Its Association with Metabolic Risk Factors in a Northwest Indian Obese Population

Sadashiv,Anupama Modi,Manoj Khokhar,Praveen Sharma,Rajnish Joshi,Sudhanshu Shekhar Mishra,Rajay N Bharshankar,Sunita Tiwari,Pankaj Kumar Singh,Vivek Vidyadhar Bhosale,Mahendra Pal Singh Negi 대한비만학회 2021 The Korean journal of obesity Vol.30 No.3

Background: It is well established that obesity is a major health risk in diabetes and associated diseases. Epigenetic changes, specially DNA methylation, play an important role in regulation of adipokines. The objective of the present study was to evaluate the DNA methylation status at the promoter region of the leptin gene in obese individuals and its association with metabolic risk factors. Methods: The study included obese (n=100) and non-obese (n=75) individuals aged 25–45 years, and measured their physical, biochemical parameters (glucose, insulin, and lipid profiles) and leptin, DNA methyltransferase 1 (DNMT1), and DNA methyltransferase 3 beta (DNMT3b) mRNA expressions with real-time reverse transcription-polymerase chain reaction (qRT-PCR). DNA methylation of the leptin gene at the promoter region was analyzed by methyl-specific qPCR . Results: The study found that the DNA methylation level at the promoter area of the leptin gene was negatively associated with weight in obese subjects. Furthermore, study findings showed that the DNA methylation level was negatively associated with fasting insulin, glucose, homeostatic model assessment for insulin resistance, and total cholesterol. There was also a higher expression of DNMT1 and DNMT-3b in obese subjects as compared with non-obese subjects. Conclusion: The leptin epigenetic profile may be associated with obesity and its associated metabolic risk factors.

Comparative Biochemical Characterization of Three Exolytic Oligoalginate Lyases from <i>Vibrio splendidus</i> Reveals Complementary Substrate Scope, Temperature, and pH Adaptations

Jagtap, Sujit Sadashiv,Hehemann, Jan-Hendrik,Polz, Martin F.,Lee, Jung-Kul,Zhao, Huimin American Society for Microbiology 2014 Applied and environmental microbiology Vol.80 No.14

<P>Marine microbes use alginate lyases to degrade and catabolize alginate, a major cell wall matrix polysaccharide of brown seaweeds. Microbes frequently contain multiple, apparently redundant alginate lyases, raising the question of whether these enzymes have complementary functions. We report here on the molecular cloning and functional characterization of three exo-type oligoalginate lyases (OalA, OalB, and OalC) from <I>Vibrio splendidus</I> 12B01 (12B01), a marine bacterioplankton species. OalA was most active at 16°C, had a pH optimum of 6.5, and displayed activities toward poly-β-<SMALL>d</SMALL>-mannuronate [poly(M)] and poly-α-<SMALL>l</SMALL>-guluronate [poly(G)], indicating that it is a bifunctional enzyme. OalB and OalC were most active at 30 and 35°C, had pH optima of 7.0 and 7.5, and degraded poly(M·G) and poly(M), respectively. Detailed kinetic analyses of oligoalginate lyases with poly(G), poly(M), and poly(M·G) and sodium alginate as substrates demonstrated that OalA and OalC preferred poly(M), whereas OalB preferred poly(M·G). The catalytic efficiency (<I>k</I><SUB>cat</SUB>/<I>K<SUB>m</SUB></I>) of OalA against poly(M) increased with decreasing size of the substrate. OalA showed <I>k</I><SUB>cat</SUB>/<I>K<SUB>m</SUB></I> from 2,130 mg<SUP>−1</SUP> ml s<SUP>−1</SUP> for the trisaccharide to 224 mg<SUP>−1</SUP> ml s<SUP>−1</SUP> for larger oligomers of ∼50 residues, and 50.5 mg<SUP>−1</SUP> ml s<SUP>−1</SUP> for high-molecular-weight alginate. Although OalA was most active on the trisaccharide, OalB and OalC preferred dimers. Taken together, our results indicate that these three Oals have complementary substrate scopes and temperature and pH adaptations.</P>

Phytoremediation of diesel-contaminated soil and saccharification of the resulting biomass

Jagtap, Sujit Sadashiv,Woo, Seong Min,Kim, Tae-Su,Dhiman, Saurabh Sudha,Kim, Dongwook,Lee, Jung-Kul Elsevier 2014 Fuel Vol.116 No.-

<P><B>Abstract</B></P> <P>In this study, we aimed to identify plant species capable of remediating diesel-contaminated soil and to convert their biomass to bioethanol. Three plant species (<I>Pinus densiflora</I>, <I>Populus tomentiglandulosa</I>, and <I>Thuja orientalis</I>) were grown on an area of soil contaminated with 6000mg/kg of diesel to assess the effects of addition of a microbial consortium and fertilizer on remediation efficacy. Diesel-contaminated soil resulted in reduced plant biomass for most of the tested plants. However, in diesel-contaminated <I>P. densiflora</I> pots containing the microbial consortium, shoot biomass was greater than that in pots treated with diesel alone. Additionally, fertilizer application was found to be the most important factor for efficient diesel degradation. Plant biomass in diesel-contaminated soil was pretreated and used as a substrate for hydrolysis using lignocellulases from <I>Armillaria gemina</I>, a newly isolated fungal strain. The strain showed the highest β-glucosidase (15U/mL), cellobiohydrolase (34U/mL), endoglucanase (146U/mL), endoxylanase (1270U/mL), laccase (0.16U/mL), mannanase (57U/mL), lignin peroxidase (0.31U/mL) and filter paper (1.72U/mL) activities. The highest saccharification yield was obtained with <I>P. densiflora</I> (52%). The <I>A. gemina</I> enzymes hydrolyzed the woody biomass used for phytoremediation and resulted in a high level of reducing sugar (375mg/g-substrate).</P> <P><B>Highlights</B></P> <P> <UL> <LI> The three plant species are grown in 6000mg/kg diesel-contaminated soil. </LI> <LI> The addition of a microbial consortium and fertilizer increases remediation efficacy. </LI> <LI> It reports saccharification of woody biomasses using lignocellulases from <I>Armillaria gemina</I>. </LI> <LI> <I>A. gemina</I> can be a good option for reducing sugar production from woody biomasses used for phytoremediation. </LI> </UL> </P>

Characterization of a novel endo-β-1,4-glucanase from Armillaria gemina and its application in biomass hydrolysis

Jagtap, Sujit Sadashiv,Dhiman, Saurabh Sudha,Kim, Tae-Su,Kim, In-Won,Lee, Jung-Kul Springer-Verlag 2014 Applied Microbiology and Biotechnology Vol.98 No.2

Evaluation of bonding efficiency between facial silicone and acrylic resin using different bonding agents and surface alterations

Shetty, Uttam Sadashiv,Guttal, Satyabodh Shesharaj The Korean Academy of Prosthodonitics 2012 The Journal of Advanced Prosthodontics Vol.4 No.3

PURPOSE. The aim of the study was to evaluate the effect of 3 silicone primers and 3 surface characterization of acrylic resin surface on bond strength between silicone elastomer and acrylic resin. MATERIALS AND METHODS. 96 Cosmesil silicones bonded to heat-curing acrylic resin were fabricated with the dimension of $75{\times}10{\times}3$ mm. The 3 primers used in this study were G611 platinum primer, A-330 Gold platinum primer, and cyanoacrylates resin. Specimens without primer were used as control. The 3 types of surface characterization done were retentive holes with 1.5 mm in diameter and 0.5 mm deep, retentive beads of 0.6 mm diameter and the third type which was plain without any characterization. The specimens were then checked for bond strength by subjecting them to $180^{\circ}$ peel test on a universal testing machine. The obtained results were then subjected to statistical analysis using 2-way ANOVA and Scheff$\acute{e}$ multiple post hoc procedures. The statistical significance was set at 5% level of significance. RESULTS. The maximum bond strength was seen for samples in which A-330G primer was used followed by G611 primer. The control group showed the minimum bond strength. Surface characterization of retentive holes increased the bond strength considerably as compared to retentive beads and samples without any surface characterization. CONCLUSION. Within the limitations of the study, A-330G primer was more compatible with Cosmesil M511 silicone and has better bonding of Cosmesil to acrylic resin. Retentive holes made on acrylic surface increased the bond strength considerably than those without any surface characterization.

Cloning and characterization of a galactitol 2-dehydrogenase from Rhizobium legumenosarum and its application in d-tagatose production

Jagtap, Sujit Sadashiv,Singh, Ranjitha,Kang, Yun Chan,Zhao, Huimin,Lee, Jung-Kul Elsevier 2014 Enzyme and microbial technology Vol.58 No.-

<P>Galactitol 2-dehydrogenase (GDH) belongs to the protein subfamily of short-chain dehydrogenases/reductases and can be used to produce optically pure building blocks and for the bioconversion of bioactive compounds. An NAD<SUP>+</SUP>-dependent GDH from Rhizobium leguminosarum bv. viciae 3841 (RlGDH) was cloned and overexpressed in Escherichia coli. The RlGDH protein was purified as an active soluble form using His-tag affinity chromatography. The molecular mass of the purified enzyme was estimated to be 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 114 kDa by gel filtration chromatography, suggesting that the enzyme is a homotetramer. The enzyme has an optimal pH and temperature of 9.5 and 35°C, respectively. The purified recombinant RlGDH catalyzed the oxidation of a wide range of substrates, including polyvalent aliphatic alcohols and polyols, to the corresponding ketones and ketoses. Among various polyols, galactitol was the preferred substrate of RlGDH with a K<SUB>m</SUB> of 8.8 mM, k<SUB>cat</SUB> of 835 min<SUP>-1</SUP> and a k<SUB>cat</SUB>/K<SUB>m</SUB> of 94.9 min<SUP>-1</SUP> mM<SUP>-1</SUP>. Although GDHs have been characterized from a few other sources, RlGDH is distinguished from other GDHs by its higher specific activity for galactitol and broad substrate spectrum, making RlGDH a good choice for practical applications.</P>

Sialoglycoproteins of Mammalian Erythrocyte Membranes: A Comparative Study

Sharma, Savita,Gokhale, Sadashiv M. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.12

The presence of sialoglycoproteins (SGPs) in the membranes from goat (Capra aegagrus hircus), buffalo (Bubalus bubalis bubalis) and pig (Sus scrofa domestica) erythrocytes was investigated by partial purification with a chloroform-methanol extraction method followed by Sodium dodecyl sulphate - Polyacrylamide gel electrophoresis in comparison to human (Homo sapiens) erythrocytes. The results show that mammalian erythrocytes possess clear differences in the SGPs numbers and molecular weights although all animals studied in this experiment are from the same class i.e. mammalia. The SGPs number in human, goat, buffalo and pig are four (PAS-1 to PAS-4), ten (PAS-GI to PAS-GX), seven (PAS-BI to PAS-BVII) and four (PAS-PI to PAS-IV) respectively as indicated by staining the polyacrylamide gel with sialoglycoprotein-specific Periodic acid-Schiff's (PAS) stain. The new SGPs could be observed only after the partial purification of membrane fractions named as PAS-HI with molecular weight (Mr) 190 kDa and PAS-HII 150 kDa in human, PAS-BIA in buffalo and PAS-PIA and PAS-PIVA in pig. The gels were also stained with Coomassie brilliant blue (CBB) and Silver stain to check the contamination of other membrane proteins in the purified fractions. The quantitative distribution of SGPs was also determined by densitometry. Present study indicates that there are some basic differences in mammalian erythrocyte membrane SGPs, especially with respect to their number and molecular weights indicating major structural variations.

Alginate Lyases from Alginate-Degrading <i>Vibrio splendidus</i> 12B01 Are Endolytic

Badur, Ahmet H.,Jagtap, Sujit Sadashiv,Yalamanchili, Geethika,Lee, Jung-Kul,Zhao, Huimin,Rao, Christopher V. American Society for Microbiology 2015 Applied and environmental microbiology Vol.81 No.5

<P>Alginate lyases are enzymes that degrade alginate through β-elimination of the glycosidic bond into smaller oligomers. We investigated the alginate lyases from <I>Vibrio splendidus</I> 12B01, a marine bacterioplankton species that can grow on alginate as its sole carbon source. We identified, purified, and characterized four polysaccharide lyase family 7 alginates lyases, AlyA, AlyB, AlyD, and AlyE, from <I>V. splendidus</I> 12B01. The four lyases were found to have optimal activity between pH 7.5 and 8.5 and at 20 to 25°C, consistent with their use in a marine environment. AlyA, AlyB, AlyD, and AlyE were found to exhibit a turnover number (<I>k</I><SUB>cat</SUB>) for alginate of 0.60 ± 0.02 s<SUP>−1</SUP>, 3.7 ± 0.3 s<SUP>−1</SUP>, 4.5 ± 0.5 s<SUP>−1</SUP>, and 7.1 ± 0.2 s<SUP>−1</SUP>, respectively. The <I>K<SUB>m</SUB></I> values of AlyA, AlyB, AlyD, and AlyE toward alginate were 36 ± 7 μM, 22 ± 5 μM, 60 ± 2 μM, and 123 ± 6 μM, respectively. AlyA and AlyB were found principally to cleave the β-1,4 bonds between β-<SMALL>d</SMALL>-mannuronate and α-<SMALL>l</SMALL>-guluronate and subunits; AlyD and AlyE were found to principally cleave the α-1,4 bonds involving α-<SMALL>l</SMALL>-guluronate subunits. The four alginate lyases degrade alginate into longer chains of oligomers.</P>

Experimental investigation of mixing in a novel continuous chaotic mixer

Seyyed Mostafa Hosseinalipour,Amir Tohidi,Payam Rahim Mashaei,Arun Sadashiv Mujumdar 한국화학공학회 2014 Korean Journal of Chemical Engineering Vol.31 No.10

This paper presents and discusses results of an experimental study of laminar mixing of a highly viscousfluid (dough) in a continuous chaotic mixer. The mixer consists of an eccentric rotor that rotates co-axially within astator, which results in chaotic advection. A dye injection technique was used to measure the mixing performance ofthe mixer. A mixing index was defined and computed by image processing of photographs of the exiting fluid fromthe mixer. Mixing characteristics were determined for constant as well as stepwise rotation of the rotor. Results revealedthat mixing performance improves with increase in the rotational speed for constant rotational speed. The stepwiserotation case displayed better mixing performance than the constant speed case for stepwise changes of the amplitude aswell as frequency of rotation.

VIEKIRA PAK associated drug-induced interstitial lung disease: Case series with systematic review of literature

Yu Jun Wong,Si Yuan Chew,John Chen Hsiang,Prem Harichander Thurairajah,Rahul Kumar,Eng Kiong Teo,Roshni Sadashiv Gokhale,Imran Bin Mohamed Noor,Jessica Tan 대한간학회 2019 Clinical and Molecular Hepatology(대한간학회지) Vol.25 No.2