• Title/Summary/Keyword: Hormone Analysis

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DnaJC18, a Novel Type III DnaJ Family Protein, is Expressed Specifically in Rat Male Germ Cells

  • Gomes, Cynthia;Soh, Jaemog
    • Development and Reproduction
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    • v.21 no.3
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    • pp.237-247
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    • 2017
  • Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The full-length DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

Lipase Diversity in Glacier Soil Based on Analysis of Metagenomic DNA Fragments and Cell Culture

  • Zhang, Yuhong;Shi, Pengjun;Liu, Wanli;Meng, Kun;Bai, Yingguo;Wang, Guozeng;Zhan, Zhichun;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.888-897
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    • 2009
  • Lipase diversity in glacier soil was assessed by culture-independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partiallipases shared 51-82% identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above-mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of ${\le}40^{\circ}C$. Our study indicates the abundant lipase diversity in glacier soil as well as the feasibility of sequence-based screening in discovering new lipase genes from complex environmental samples.

Prediction of Non-Genotoxic Carcinogenicity Based on Genetic Profiles of Short Term Exposure Assays

  • Perez, Luis Orlando;Gonzalez-Jose, Rolando;Garcia, Pilar Peral
    • Toxicological Research
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    • v.32 no.4
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    • pp.289-300
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    • 2016
  • Non-genotoxic carcinogens are substances that induce tumorigenesis by non-mutagenic mechanisms and long term rodent bioassays are required to identify them. Recent studies have shown that transcription profiling can be applied to develop early identifiers for long term phenotypes. In this study, we used rat liver expression profiles from the NTP (National Toxicology Program, Research Triangle Park, USA) DrugMatrix Database to construct a gene classifier that can distinguish between non-genotoxic carcinogens and other chemicals. The model was based on short term exposure assays (3 days) and the training was limited to oxidative stressors, peroxisome proliferators and hormone modulators. Validation of the predictor was performed on independent toxicogenomic data (TG-GATEs, Toxicogenomics Project-Genomics Assisted Toxicity Evaluation System, Osaka, Japan). To build our model we performed Random Forests together with a recursive elimination algorithm (VarSelRF). Gene set enrichment analysis was employed for functional interpretation. A total of 770 microarrays comprising 96 different compounds were analyzed and a predictor of 54 genes was built. Prediction accuracy was 0.85 in the training set, 0.87 in the test set and increased with increasing concentration in the validation set: 0.6 at low dose, 0.7 at medium doses and 0.81 at high doses. Pathway analysis revealed gene prominence of cellular respiration, energy production and lipoprotein metabolism. The biggest target of toxicogenomics is accurately predict the toxicity of unknown drugs. In this analysis, we presented a classifier that can predict non-genotoxic carcinogenicity by using short term exposure assays. In this approach, dose level is critical when evaluating chemicals at early time points.

Differences of Experts and Non-experts in Perceiving Environmental and Technological Risks (전문가와 비전문가의 환경 및 과학기술 위험에 대한 위해도 인식 차이)

  • Hahm, Myung-Il;Kwon, Ho-Jang;Lee, Hoo-Yeon;Park, Hwa-Gyoo;Lee, Sang-Gyu
    • Journal of Environmental Health Sciences
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    • v.35 no.4
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    • pp.269-277
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    • 2009
  • Objective : The objective of this study was to examine how experts and the public perceived various environmental and technological hazards based on psychometric paradigm. Methods : We conducted a survey that included 30 hazards and 10 risk attributes. Subjects of this study were 214 people with three groups; (1) experts (55 people), (2) graduates( 78 people), (3) under graduates (81 people). Factor analysis was used to confirm the common risk attribute from 8 risk attributes. Also, multiple regression analysis was used to identify factors influencing on perceived risk and benefit of hazards. Results : This study revealed that experts tend to be more tolerant than graduates and under graduate students for the 30 hazards. Using factor analysis, two main factors were identified: factor 1, commonly called "Dread Risk", and factors 2, commonly called, "Unknown Risk" in the literature. We identified that environmental hormone concentration and global warming ranked high in both dread risk and unknown risk. Multiple regression models were used to test the association of perceived social risk and perceived social benefit with two main factors. Dread risk had significant explanatory power on perceived social risk and benefit. We identified that the experts were less likely to perceived dread risk and know more information about the hazards. Conclusions : There were differences of risk perception between experts and lay people. Especially, experts' perception of risk was commonly lower than other people's perception.

Growth Inhibition and Apoptosis Induction of Essential Oils and Extracts of Nepeta cataria L. on Human Prostatic and Breast Cancer Cell Lines

  • Emami, Seyed Ahmad;Asili, Javad;HosseinNia, Shima;Yazdian-Robati, Rezvan;Sahranavard, Mehrdad;Tayarani-Najaran, Zahra
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.125-130
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    • 2016
  • Nepeta cataria L. has been used in traditional medicine of some countries. Here the cytotoxic and apoptogenic activity of methanol extracts, n-hexane, dichloromethane, ethyl acetate, n-butanol, and acqueous extracts and the essential oil obtained from the aerial parts of the plant were evaluated with PC3, DU-145 and MCF-7 cell lines. Cell viability, histograms of PI stained fragmented DNA in apoptotic cells and Western blot analysis of proteins involved in the cascade of apoptosis were compared in all samples. Thirty components were identified as volatile, representing 99.7% of essential oil composition after GC-MS analysis of the oil obtained from aerial parts of the N. cataria by hydro-distillation. The major oil components of the essential oil were nepetalactone stereoisomers. Comparing IC50 values showed estrogen receptor positive PC3 cells were more sensitive to the cytotoxic effects of N. cataria in comparison with low hormone-receptor presenting DU-145 cells. Among multiple extracts and essential oils of the plant, only the ethyl acetate extract could significantly decrease cell viability in PC3 cells, in a concentration dependent manner. Ethyl acetate extract of N. cataria treated cells showed a sub-G1 peak in PC3 cells in a concentration dependent manner that indicates the involvement of an apoptotic process in ethyl acetate extract-induced cell death. Western blotting analysis showed that in PC3 cells treated with ethyl acetate (48 h) caspase 3 and PARP were cleaved to active forms. Overall, the results suggest that further analytical elucidation of N. cataria in respect to finding new cytotoxic chemicals with anti-tumor activity is warranted.

Content Analysis of Questions Related to Breast Cancer Raised through Internet Counseling in Korea (유방암 관련 인터넷 상담 질문에 대한 내용분석)

  • Yi, Myung-Sun;Noh, Dong-Young;Kim, Kum-Ja;Yih, Bong-Sook
    • Asian Oncology Nursing
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    • v.7 no.2
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    • pp.119-130
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    • 2007
  • Purpose: The purpose of this study was to analyze the data regarding questions raised by women with breast cancer through Internet counseling in Korea. Methods: The data were collected from one internet web-site, providing counseling by physicians. A total of 617 questions were analyzed by content analysis method. Results: About 90 percent of the counselees were patients themselves. But most of the general and health-related characteristics of them were not known from the data. As a result of content analysis, 617 questions were grouped into 9 major categories. The most common major category was identified as "life after treatment" (212 questions, 34.2%), followed by "chemotherapy" (139 questions, 22.3%) and "hormone therapy" (115 questions, 18.9%). Questions regarding "physical symptoms" were the most frequent one in the major categories of "life after treatment", "chemotherapy", and "radiotherapy", while questions regarding "psychological problems" were the least. Conclusion: The findings of this study indicate that it is important for health professionals to provide continuous on-line informational support to women with breast cancer, even after all the treatment is over, especially focusing on physical symptoms. In addition, off-line program needs to be reinforced to provide emotional support that is not well delivered by on-line program.

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Three cases of rare SRY-negative 46,XX testicular disorder of sexual development with complete masculinization and a review of the literature

  • Lee, Bom Yi;Lee, Shin Young;Lee, Yeon Woo;Kim, Shin Young;Kim, Jin Woo;Ryu, Hyun Mee;Lee, Joong Shik;Park, So Yeon;Seo, Ju Tae
    • Journal of Genetic Medicine
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    • v.13 no.2
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    • pp.78-88
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    • 2016
  • Purpose: To identify the clinical characteristics of SRY-negative male patients and genes related to male sex reversal, we performed a retrospective study using cases of 46,XX testicular disorders of sex development with a review of the literature. Materials and Methods:SRY-negative cases of 46,XX testicular disorders of sex development referred for cytogenetic analysis from 1983 to 2013 were examined using clinical findings, seminal analyses, basal hormone profiles, conventional cytogenetic analysis and polymerase chain reaction. Results: Chromosome analysis of cultured peripheral blood cells of 8,386 individuals found 19 cases (0.23%) with 46,XX testicular disorders of sex development. The SRY gene was confirmed to be absent in three of these 19 cases (15.8%). Conclusion: We report three rare cases of SRY-negative 46,XX testicular disorders of sex development. Genes on autosomes and the X chromosome that may have a role in sex determination were deduced through a literature review. These genes, through differences in gene dosage variation, may have a role in sex reversal in the absence of SRY.

Effects of scalp treatment using combinational massage technique on human physiology

  • Oh, Gang-Su;Kim, Sung-Nam
    • Journal of Fashion Business
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    • v.12 no.3
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    • pp.87-98
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    • 2008
  • The purposes of this study are to measure physiological reactions of human body according to the scalp treatment, a popularized service in beauty care industry and propose efficient ways of scalp treatment. To meet the goals, total 30 applicants without any medical history(5 males and 5 females in 20's, 30's and 40's respectively) were informed on the purpose of experiment hereof and were investigated and received a 30-minute scalp treatment, which combines standardized scalp treatment massage technique proposed by KAT and ITF with another massage technique operated in the beauty salon run by the author of this paper. 5ml of blood samples were taken from each subject before and after the scalp treatment respectively and the blood sample was divided into 3 different tubes for analysis: 1) 2 ml for blood cell analysis, 2) 2ml for enzyme activity measurement, 3) 1ml for hormone level reading. In order to determine effects of scalp treatment on ALP, GOT, GPT, ${\gamma}-GTP$, WBC, RBC, Hb, Hct, Platelet, MCV, MCH and MCHC, all collected data were used for measuring respective levels of these blood substances by means of enzyme reaction measurement, enzyme activity measurement and automated hematology analyzer. Then, all measured data were analyzed through paired t-test using SPSS WIN 11.5. As a result, the scalp treatment is associated with improving hepatic function, facilitating blood circulation and helping blood coagulation and hemostasis in a effective way. Therefore, it would be necessary to conduct further studies on this subject related to anemia in the future.

Isolation and Characterization of Plant Growth Promoting Rhizobacteria from Waste Mushroom bed from Agaricus bisporus (양송이 수확 후 배지로부터 식물생장촉진세균의 분리 및 생육특성)

  • Jung, Young-Pil;Kyung, Ki-Cheon;Jang, Kab-Yeul;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.866-871
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    • 2011
  • An auxin-producing bacteria (3YN11-02) was isolated from waste mushroom bed from Agaricus bisporus of Chungnam Buyeo-Gun area. The strain 3YN11-02 was identified as a novel species belongs to Rahnella aquatica by a chemotaxanomic and phylogenetic nalysis. The isolate was confirmed to produce indole-3-acetic acid (IAA) which is one of auxin hormone by TLC and HPLC analysis. When the concentration of IAA was assessed by performing HPLC quantity analysis, the maximal $290mg\;L^{-1}$ of IAA detected in ether fraction extracted from the culture filtrate which was cultured in R2A broth containing 0.1% tryptophan for 24 h at $35^{\circ}C$. The molecular weight of the main peak obtained by LC-mass analysis was correspondent well to 175, that of IAA. To investigate the growth promoting effect of crop, when the culture broth of R. aquatica 3YN11-02 was infected onto water culture and seed pot of mung bean, the adventitious root induction and root growth of mung bean were 2.0 times higher than control.

Transcriptome profiling and identification of functional genes involved in H2S response in grapevine tissue cultured plantlets

  • Ma, Qian;Yang, Jingli
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1287-1300
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    • 2018
  • Hydrogen sulfide ($H_2S$), a small bioactive gas, has been proved functioning in plant growth and development as well as alleviation of abiotic stresses, which including promoting seed germination, accelerating embryonic root growth, regulating flower senescence, inducing stomatal closure, and defending drought, heat, heavy metals and osmotic stresses etc. However, the molecular functioning mechanism of $H_2S$ was still unclear. The primary objective of this research was to analyze the transcriptional differences and functional genes involved in the $H_2S$ responses. In details, 4-week-old plantlets in tissue culture of grapevine (Vitis vinifera L.) cultivar 'Zuoyouhong' were sprayed with 0.1 mM NaHS for 12 h, and then transcriptome sequencing and qRT-PCR analysis were used to study the transcriptional differences and functional genes involved in the $H_2S$ responses. Our results indicated that 650 genes were differentially expressed after $H_2S$ treatment, in which 224 genes were up-regulated and 426 genes were down-regulated. The GO enrichment analysis and KEGG enrichment analysis results indicated that the up-regulated genes after $H_2S$ treatment focused on carbon metabolism, biosynthesis of amino acids, and glycolysis/gluconeogenesis, and the down-regulated genes were mainly in metabolic pathways, biosynthesis of secondary metabolites, and plant hormone signal transduction. Analyzing the transcription factor coding genes in details, it was indicated that 10 AP2/EREBPs, 5 NACs, 3 WRKYs, 3 MYBs, and 2 bHLHs etc. transcription factor coding genes were up-regulated, while 4 MYBs, 3 OFPs, 3 bHLHs, 2 AP2/EREBPs, 2 HBs etc. transcription factor coding genes were down-regulated. Taken together, $H_2S$ increased the productions in secondary metabolites and a variety of defensive compounds to improve plant development and abiotic resistance, and extend fruits postharvest shelf life by regulating the expression of AP2/EREBPs, WRKYs, MYBs, CABs, GRIP22, FERRITINs, TPSs, UGTs, and GHs etc.