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Journal of Bioresources and Bioproducts 2023年第2期8卷 108-124页

作者： Akansha Shrivastava Mamta Pal rakesh Kumar sharma Department of Biosciences Manipal University JaipurJaipur 303007India Sardar Swaran Singh National Institute of Bio-Energy Jalandhar 144601India

Yeast has been used as a cell factory for thousands of years to produce a wide variety of com-plex biofuels,bioproducts,biochemicals,food ingredients,and *** a variety of biotechnological production hosts,a few specific genera of yeast have proven *** developments in metabolic engineering and synthetic biology provide a workable long-term supply solution for these *** this review,we have covered recent advances in the design of yeast cell factories for the synthesis of terpenoids,alkaloids,phenylpropanoids,and other natural chemicals,primarily focusing on Pichia ***-edge solutions involving genetic and process engineering have also been ***,the review summarized recent advancements and challenges in synthetic and systems biology,as well as initiatives in metabolic engineering aimed at commercializing non-conventional yeasts like *** processes used in non-traditional yeasts to produce enzymes,therapeutic proteins,lipids,and metabolic products for industrial applications were thoroughly elaborated.

关键词： Natural product Pichia Non-conventional yeast Biosynthesis System biology Synthetic biology

Proteomic analysis of mature and immature ejaculated spermatozoa from fertile men

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Asian Journal of Andrology 2016年第5期18卷 735-746页

作者： Zhihong Cui rakesh sharma Ashok Agarwal American Center for Reproductive Medicine Cleveland Clinic Cleveland OH 44195 USA Institute of Toxicology Third Military Medical University Chongqing 400038PR China.

Dysfunctional spermatozoa maturation is the main reason for the decrease in sperm motility and morphology in infertile men. Ejaculated spermatozoa from healthy fertile men were separated into four fractions using three-layer density gradient. Proteins were extracted and bands were digested on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Functional annotations of proteins were obtained using bioinformatics tools and pathway databases. Western blotting was performed to verify the expression levels of the proteins of interest. 1469 proteins were identified in four fractions of spermatozoa. The number of detected proteins decreased according to the maturation level of spermatozoa. During spermatozoa maturation, proteins involved in gamete generation, cell motility, energy metabolism and oxidative phosphorylation processes showed increasing expression levels and those involved in protein biosynthesis, protein transport, protein ubiquitination, and response to oxidative stress processes showed decreasing expression levels. We validated four proteins （HSP 70 1A, clusterin, tektin 2 and tektin 3） by Western blotting. The study shows protein markers that may provide insight into the ejaculated spermatozoa proteins in different stages of sperm maturation that may be altered or modified in infertile men.

关键词： bioinformatics protein validation proteins proteomics spermatozoa maturation

Proteomic analysis reveals dysregulated cell sign aling in ejaculated spermatozoa from in fertile men

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Asian Journal of Andrology 2019年第2期21卷 121-130页

作者： Luna Samanta rakesh sharma Zhihong Cui Ashok Agarwal American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Zoology School of Life SciencesRavenshaw UniversityCuttackOdisha 753003India Institute of Toxicology The Third Military Medical UniversityChongqing 400038China

Dysfunctional sperm maturation is the primary reason for the poor sperm motility and morphology in infertile *** from infertile men were fractioned on three-layer density gradient(80%,60%,and 40%).Fraction 1(Fl)refers to the least mature stage having the lowest den sity,whereas the fraction 4(F4)in eludes the most dense and morphologically mature motile *** 2(F2)and fraction 3(F3)represent the intermediate *** were extracted and separated by dimensional *** were digested with trypsin and analyzed on a LTQ-Orbitrap Elite hybrid mass spectrometer *** annotations of proteins were obtained using bioinformatics tools and pathway databases.A total of 1585 proteins were detected in the four fractions of spermatozoa.A dysregulated protein turnover and protein folding may lead to accumulation of defective proteins or proteins that otherwise would have been eliminated during the process of maturation,resulting in the impairment of sperm fun *** nt chaper one expressi on may be a major con tributing factor to the defective sperm *** n receptor was predicted as a transcription regulator in one of the networks and the affected pathways were chaperone-mediated stress response,proteosomal pathway,and sperm *** down regulation of key pathways and protei ns which compromises the fertilizi ng potential of spermatozoa may provide in sight into the mecha nisms that lead to male infertility.

关键词： androgen receptor chaperone immature sperm infertile men proteasome spermatozoa

Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation:an appeal for performance verification

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Asian Journal of Andrology 2019年第5期21卷 438-444页

作者： rakesh sharma Sajal Gupta Ralf Henkel Ashok Agarwal American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Medical Bioscience University of the Western CapeBellville 7535South Africa

Lack of standardized,reproducible protocols and referencO values is among the challenges faced when using new or upgraded versions of instruments in reproductive laboratories and flow *** deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay combined with flow cytometry routinely used for diagnostic measurement of sperm DNA fragmentation(SDF)is a unique *** change in the setting of the standard instrument,including upgrades of hardware or software,can lead to different results and may affect clinicians'decision for ***,we compared TUNEL results of SDF obtained from a standard(C6)flow cytometer with a newer version of the same instrument(C6 Plus)and examined the cutoff,sensitivity,and specificity without calibration(adjustment)and after *** sperm preparation and matched acquisition settings were used to examine the performance of two flow *** strength of agreement of the results between the two observers was also *** adjustment of the settings,overall concordance became high and the two cytometers showed 100%positive and negative predictive value with 100%area under the *** overall correlation coefficient observed between C6 and C6 Plus was highly significant(P99%.The in terobserver agreeme nt on C6 flow cytometer for the two observers was 0.801±0.062 and 0.746±0.044 for C6 *** demonstrated a strong agreement between the samples tested on the two flow cytometers after calibration and established the robustness of both instruments.

关键词： DNA fragmentation flow cytometer oxidative stress sperm TUNEL assay

Vigna radiata var. GM4 Plant Growth Enhancement and Root Colonization by a Multi-Metal-Resistant Plant Growth-Promoting Bacterium Enterobacter sp. C1D in Cr(VI)-Amended Soils

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Pedosphere 2018年第1期28卷 144-156页

作者： Gangavarapu SUBRAHMANYAM rakesh Kumar sharma Gattupalli Naresh KUMAR Gattupalli ARCHANA Department of Microbiology and Biotechnology Centre The Maharaja Sayajirao University of Baroda Department of Biomedical Science Nitte University Center for Science Education and Research (NUCSER) Nitte University Central Muga Eri Research and Training Institute Central Silk Board Department of Biochemistry Faculty of Science The Maharaja Sayajirao University of Baroda

Contamination of agricultural soils by heavy metals has become a major concern due to their toxic effects on plant growth,symbiosis and consequently the yields of crops. In the present study, to enhance plant growth in Cr（VI）-amended soils, novel metalresistant plant growth-promoting bacteria（PGPB） were isolated from a soil contaminated with industrial waste effluent. One of the bacterial isolates, identified as Enterobacter sp. C1 D by 16 S r RNA gene sequencing, was found to be multi-metal resistant in nature with excellent plant growth-promoting（PGP） traits. Mung bean（Vigna radiata var. GM4） inoculation with Enterobacter sp.C1 D significantly（P ＆lt; 0.01） increased root and shoot length, shoot and root weight, and chlorophyll content in a range of Cr（VI）treatments. Plant tolerance towards Cr（VI） measured as effective concentration showed higher values with Enterobacter sp. C1 Dtreated plants compared to un-inoculated plants. Root colonization study was also carried out using green fluorescence protein-labeled Enterobacter sp. C1 D under a hydroponic system. Confocal laser scanning microscopy of the plant roots showed heavy bacterial loads on the surface of the plant root specifically at the root tip and the point of root hair/lateral root formation. The results of PGP traits showed that elevated indole acetic acid levels and 1-aminocyclopropane-1-carboxylate deaminase activity enabled Enterobacter sp. C1 D to enhance V. radiata growth in Cr（VI）-amended soils, whereby it significantly increased plant tolerance towards elevated Cr（VI） concentrations.

关键词： chromium(VI) Enterobacter sp C1D heavy metal metal toxicity plant tolerance

Proteomic signatures of infertile men with clinical varicocele and their validation studies reveal mitochondrial dysfunction leading to infertility

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Asian Journal of Andrology 2016年第2期18卷 282-291,J0007页

作者： Ashok Agarwal rakesh sharma Luna Samanta Damayanthi Durairajanayagam Edmund Sabanegh American Center for Reproductive Medicine Cleveland Clinic Cleveland OH USA Department of Zoology School of Life Sciences Ravenshaw UniversityCuttack - 751 003 Odisha India Faculty of Medicine MARA University of Technology (UiTM) Selangor Darul Ehsan Malaysia Department of Urology Cleveland Clinic Cleveland OH USA.

To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each varicocele outcome were combined to identify the proteins involved in varicocele-associated infertility in men irrespective of stage and laterality of their clinical varicocele. Expression levels of 5 key proteins （PKARIA, AK7, CCT6B, HSPA2, and ODF2） involved in stress response and sperm function including molecular chaperones were validated by Western blotting. Ninety-nine proteins were differentially expressed in the varicocele Eroup. Over 87% of the DEP involved in major energy metabolism and key sperm functions were underexpressed in the varicocele group. Key protein functions affected in the varicocele group were spermatogenesis, sperm motility, and mitochondrial dysfunction, which were further validated by Western blotting, corroborating the proteomics analysis. Varicocele is essentially a state of energy deprivation, hypoxia, and hyperthermia due to impaired blood supply, which is corroborated by down-regulation of lipid metabolism, mitochondrial electron transport chain, and Krebs cycle enzymes. To corroborate the proteomic analysis, expression of the 5 identified proteins of interest was validated by Western blotting. This study contributes toward establishing a biomarker ＂fingerprint＂ to assess sperm quality on the basis of molecular parameters.

关键词： bioinformatics male infertility proteomics spermatozoa proteins varicocele

Strategies to mitigate the adverse effect of drought stress on crop plants-influences of soil bacteria:A review

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Pedosphere 2021年第3期31卷 496-509页

作者： Priyanka CHANDRA Amoolya WUNNAVA Pooja VERMA Amaresh CHANDRA rakesh Kumar sharma Indian Council of Agricultural Research(ICAR) Central Soil Salinity Research InstituteKarnalHaryana 132001India Department of Biosciences Manipal University JaipurRajasthanJaipur 303007India Indian Council of Agricultural Research(ICAR) Indian Institute of Sugarcane ResearchUttar PradeshLucknow 226002India

Drought stress affects plant growth and causes significant issues in meeting global demand for food crops and *** can cause physiological,physicochemical,and morphological changes in plants,which negatively affects plant growth and *** combat this under the increasing global threat of water shortage and rapid population expansion,it is crucial to develop strategies to meet global food *** growth-promoting rhizobacteria(PGPR)may provide a safe solution to enhancing crop yields through various *** soil bacteria can provide drought tolerance to crop plants,allowing them to survive and thrive in water-scarce *** of phytohormones,free radical-scavenging enzymes,and stress-combating enzymes that can increase tolerance to drought-induced stress are key features of plant-associated microbial *** review summarizes the beneficial properties of microbes that help plants tolerate water scarcity and highlights the bacterial mechanisms that enhance drought tolerance in plants.

关键词： abiotic stress agricultural yield drought tolerance food crops microbe PGPR rhizobactoria water deficit

Reduced semen quality in patients with testicular cancer seminoma is associated with alterations in the expression of sperm proteins

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Asian Journal of Andrology 2020年第1期22卷 88-93页

作者： Tania R Dias Ashok Agarwal Peter N Pushparaj Gulfam Ahmad rakesh sharma American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Health Sciences Faculty of Health SciencesUniversity of Beira InteriorCovilha 6201-001Portugal Department of Microscopy and Unit for Multidisciplinary Research in Biomedicine Institute of Biomedical Sciences Abel SalazarUniversity of PortoPorto 4050-313Portugal Center of Excellence in Genomic Medicine Research Faculty of Applied Medical SciencesJeddah 21589Saudi Arabia Division of Pathology School of Medical SciencesSydney UniversityLidcombe NSW 2141Australia

Testicular cancer seminoma is one of the most common types of cancer among men of reproductive *** with this condition usually present reduced semen quality,even before initiating cancer ***,the underlying mechanisms by which testicular cancer seminoma affects male fertility are largely *** aim of this study was to investigate alterations in the sperm proteome of men with seminoma undergoing sperm banking before starting cancer therapy,in comparison to healthy proven fertile men(control group).A routine semen analysis was conducted before cryopreservation of the samples(n=15 per group).Men with seminoma showed a decrease in sperm motility(P=0.019),total motile count(P=0.001),concentration(P=0.003),and total sperm count(P=0.001).Quantitative proteomic analysis identified 393 differentially expressed proteins between the study *** proteins involved in spermatogenesis,sperm function,binding of sperm to the oocyte,and fertilization were selected for validation by western *** confirmed the underexpression of heat shock-related 70 kDa protein 2(P=0.041),ubiquinol-cytochrome C reductase core protein 2(P=0.026),and testis-specific sodium/potassium-transporting ATPase subunit alpha-4(P=0.016),as well as the overexpression of angiotensin I converting enzyme(P=0.005)in the seminoma *** altered expression levels of these proteins are associated with spermatogenesis dysfunction,reduced sperm kinematics and motility,failure in capacitation and *** findings of this study may explain the decrease in the fertilizing ability of men with seminoma before starting cancer therapy.

关键词： male fertility proteomics seminoma sperm proteins sperm quality testicular cancer

Job satisfaction and organizational commitment among nurses working on temporary versus permanent jobs at a tertiary care teaching hospital,Uttarakhand,India

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Journal of Integrative Nursing 2022年第4期4卷 224-230页

作者： Neha PANCHAL Suresh Kumar sharma rakesh sharma Ritu RANI Department of Neurosurgery Vardhman Mahavir Medical College and Safdarjung HospitalNew DehliIndia College of Nursing All India Institute of Medical SciencesJodhpurRajasthanIndia College of Nursing All India Institute of Medical SciencesRishikeshUttarakhandIndia

Objective:The objective of this study was to assess and compare job satisfaction and organizational commitment among nurses working on temporary versus permanent basis at a tertiary care teaching hospital,Uttarakhand,*** and Methods:A total of 250 nurses(125 temporary and 125 permanent)were selected using stratified disproportionate random sampling techniques from a tertiary care teaching *** datasheet,Job Satisfaction Survey scale,and Organizational Commitment scale were used to collect ***:The study results revealed that the job satisfaction score was significantly higher in permanent nurses(139.1±15.4)than in temporary nurses(131.3±12.2)(P<0.001).However,temporary nurses(85.2±14.4)were more committed toward organization than permanent nurses(80.0±16.0)(P<0.001).There was a significant association between nurses’job satisfaction and their gross salary(P<0.001)and their organizational commitment with the selected profession by own will(P<0.001).Conclusion:The present study concluded that majority of temporary and permanent nurses reported a moderate level of job satisfaction and organizational *** study suggests enhancing employee satisfaction can improve organizational *** well,this study’s findings can serve as a valuable foundation for designing policies to enhance organizational commitment in the future.

关键词： Health-care services job satisfaction organizational commitment permanent nurses temporary nurses

Novel association between sperm deformity index and oxidative stress-induced DNA damage in infertile male patients

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Asian Journal of Andrology 2005年第2期7卷 121-126,页

作者： Tamer M.Said Nabil Aziz rakesh K.sharma Iwan Lewis-Jones AnthonyJ.ThomasJr AshokAgarwal Center for Advanced Research in Human Reproduction Infertility and Sexual FunctionGlickman Urological Institute and Department of Obstetrics-GynecologyThe Cleveland Clinic FoundationClevelandOhio 44195USA Reproduction Medicine Unit Liverpool Women's HospitalLiverpool L87SSSUK

Aim:To investigate the impact of abnormal sperm morphology using the sperm deformity index (SDI) on reactive oxygen species (ROS) production and its correlation with sperm DNA ***:Semen samples were collected from men undergoing infertility screening (n=7) and healthy donors (n=6).Mature spermatozoa were isolated and incubated with 5 mmol/L β-nicotinamide adenine dinucleotide phosphate (NADPH) for up to 24 h to induce *** morphology was evaluated using strict Tygerberg's criteria and the *** levels and DNA damage were assessed using chemiluminescence and terminal deoxynucleotidyl transferase-mediated fluorescein- dUTP nick end labeling (TUNEL) assays,***:SDI values (median [interquartiles]) were higher in patients than donors (2 [1.8,2.1] vs.1.53 [1.52,1.58],P=0.008).Aliquots treated with NADPH showed higher ROS levels (1.22 [0.30,1.87] vs.0.39 [0.10,0.57],P=0.03) and higher incidence of DNA damage than those not treated (10 [4.69,24.85] vs.3.85 [2.58,5.10],P=0.008).Higher DNA damage was also seen following 24 h of incubation in patients compared to *** correlated with the percentage increase in sperm DNA damage following incubation for 24 h in samples treated with NADPH (r=0.7,P=0.008) and controls (r=0.58,P=0.04). Conclusion:SDI may be a useful tool in identifying potential infertile males with abnormal prevalence of oxidative stress (OS)-induced DNA *** plays a role in ROS-mediated sperm DNA damage,which appears to be more evident in infertile patients with semen samples containing a high incidence of morphologically abnormal spermatozoa.

关键词： β-nicotinamide adenine dinucleotide phosphate oxidative stress sperm deformity index sperm DNA damage