• Title/Summary/Keyword: Metaphase

Search Result 332, Processing Time 0.028 seconds

Evaluation of Chromosomal Alteration in Electrical Workers Occupationally Exposed to Low Frequency of Electro Magnetic Field (EMFs) in Coimbatore Population, India

  • Balamuralikrishnan, Balasubramanian;Balachandar, Vellingiri;Kumar, Shanmugam Suresh;Stalin, Nattan;Varsha, Prakash;Devi, Subramaniam Mohana;Arun, Meyyazhagan;Manikantan, Pappuswamy;Venkatesan, Chinnakulandhai;Sasikala, Keshavarao;Dharwadkar, Shahnaz N.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.6
    • /
    • pp.2961-2966
    • /
    • 2012
  • Extremely low frequency electro magnetic fields (EMFs) have been classified as possibly carcinogenic to humans by the International Agency for Research on Cancer. An increased number of chromosomal alterations in peripheral lymphocytes are correlated with elevated incidence of cancer. The aim of the present study was to assess occupationally induced chromosomal damage in EMF workers exposed to low levels of radiation. We used conventional metaphase chromosome aberration (CA) analysis and the micronucleus (MN) assay as biological indicators of non ionizing radiation exposure. In the present study totally 70 subjects were selected including 50 exposed and 20 controls. Informed written consent was obtained from all participants and the study was performed in accordance with the Declaration of Helsinki and the approval of the local ethical committee. A higher degree of CA and MN was observed in exposed subjects compared to controls, the frequency of CA being significantly enhanced with long years of exposure (P<0.05). Moreover increase in CA and MN with age was noted in both exposed subjects and controls, but was significantly greater in the former. The results of this study demonstrated that a significant induction of cytogenetic damage in peripheral lymphocytes of workers occupationally exposed to EMFs in electric transformer and distribution stations. In conclusion, our findings suggest that EMFs possess genotoxic capability, as measured by CA and MN assays; CA analysis appeared more sensitive than other cytogenetic end-points. It can be concluded that chronic occupational exposure to EMFs may lead to an increased risk of genetic damage among electrical workers.

In vitro growth of mouse preantral follicles: effect of animal age and stem cell factor/insulin-like growth factor supplementation

  • Jee, Byung Chul;Kim, Jee Hyun;Park, Da Hyun;Youm, Hyewon;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.39 no.3
    • /
    • pp.107-113
    • /
    • 2012
  • Objective: To determine whether animal age impacts in vitro preantral follicle growth. Effects of hCG, stem cell factor (SCF), and/or insulin-like growth factor (IGF) supplementation in growth medium were also investigated. Methods: Intact preantral follicles were mechanically isolated from fresh ovaries of BDF1 mice and cultured in growth medium for 9 to 11 days. Surviving follicles with antrum formation were transferred to maturation medium for 14 to 18 hours. Follicle survival, antrum formation, and retrieval of metaphase II (MII) oocytes were compared among three age categories (4-5, 7-8, and 10-11 week-old). By using 7- to 8-week-old mice, preantral follicles were cultured in growth medium supplemented with hCG (0, 5, or 10 mIU/mL), SCF (50 ng/mL), IGF-1 (50 ng/mL), and SCF+IGF-1. Results: Seven- to eight-week-old mice showed a higher follicle survival and antrum formation and produced more MII oocytes compared to other groups. In the 7- to 8-week-old mice, supplementation of 5 mIU/mL hCG significantly enhanced the antrum formation but the percentage of MII oocytes was similar to that of the control. Supplementation of SCF+IGF-1 did not enhance follicle survival or antrum formation but the percentage of MII oocytes increased modestly (39.1%) than in the control (28.6%, p>0.05, statistically not significant). Conclusion: Seven- to eight-week-old mice showed better outcomes in growth of preantral follicles in vitro than 4- to 5- or 10- to 11-week-old mice. Supplementation of hCG enhanced antrum formation and supplementation of SCF+IGF-1 yielded more mature oocytes; hence, these should be considered in the growth of preantral follicles in vitro.

Do spontaneously decreasing estradiol levels prior to triggering of ovulation adversely impact in vitro fertilization outcomes?

  • Grin, Leonti;Berkovitz-Shperling, Roza;Zohav, Eyal;Namazov, Ahmet;Leyetes, Sophia;Friedler, Shevach
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.47 no.3
    • /
    • pp.213-220
    • /
    • 2020
  • Objective: The aim of this study was to explore the potential adverse effect of spontaneously decreasing serum estradiol (SE) levels on in vitro fertilization (IVF) outcomes. Methods: This retrospective single-subject study analyzed IVF cycles conducted at a hospital IVF unit between 2010 and 2017. Overall, 2,417 cycles were analyzed. Only cycles with spontaneously decreasing SE before human chorionic gonadotropin (hCG) triggering were included. Each patient served as her own control, and subsequent cycles were analyzed for recurrent SE decreases. The main outcome was the number of oocytes retrieved. Results: Cycle characteristics were similar between the study (SE decrease) and control groups, with the exception of the median SE on the day of hCG triggering (899.7 pg/mL; interquartile range [IQR], 193-2,116 pg/mL vs. 1,566.8 pg/mL; IQR, 249-2,970 pg/mL; p< 0.001). The study group, relative to the control group, had significantly fewer total oocytes (5 [IQR, 2-9] vs. 7 [IQR, 3-11]; p= 0.002) and significantly fewer metaphase II (MII) oocytes (3 [IQR, 1-6] vs. 4 [IQR, 2-8]; p= 0.001) retrieved. The study group had fewer cleavage-stage embryos than the control cycles (3 [IQR, 1-6] vs. 4 [IQR, 2-7]; p= 0.012). Compared to cycles with a ≤ 20% SE decrease, cycles with a > 20% decrease had significantly fewer total and MII oocytes retrieved. SE decrease recurred in 12% of patients. Conclusion: A spontaneous decrease in SE levels adversely affected IVF outcomes, with a linear correlation between the percentage decrease and the number of oocytes retrieved. SE decrease can repeat in later cycles.

Effects of Meiotic Stages, Cryoprotectants, Cooling and Vitrification on the Cryopreservation of Porcine Oocytes

  • Huang, Wei-Tung;Holtz, Wolfgang
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.15 no.4
    • /
    • pp.485-493
    • /
    • 2002
  • Different factors may affect the sensitivity of porcine oocytes during cryopreservation. The effect of two methods (cooling and vitrification), four cryoprotectants [glycerol (GLY), 1, 2-propanediol (PROH), dimethyl sulfoxide (DMSO) or ethylene glycol (EG)] and two vitrification media (1 M sucrose (SUC)+8 M EG; 8 M EG) on the developmental capacity of porcine oocytes at the germinal vesicle (GV) stage or after IVM at the metaphase II (M II) stage were examined. Survival was assessed by FDA staining, maturation and cleavage following IVF and IVC. A toxicity test for different cryoprotectants (GLY, PROH, DMSO, EG) was conducted at room temperature before cooling. GV and M II-oocytes were equilibrated stepwise in 1.5 M cryoprotectant and diluted out in sucrose. The survival rate of GV-oocytes in the GLY group was significantly lower (82%, p<0.01) than that of the other group (92 to 95%). The EG group achieved a significantly higher maturation rate (84%, p<0.05) but a lower cleavage rate (34%, p<0.01) than the DMSO group and the controls. For M II-oocytes, the survival rates for all groups were 95 to 99% and the cleavage rate of the GLY group was lower than the PROH-group (21 vs 43%, p<0.01). After cooling to $10^{\circ}C$, the survival rates of GV-oocytes in the cryoprotectant groups were 34 to 51%, however, the maturation rates of these oocytes were low (1%) and none developed after IVF. For M II-oocytes, the EG group showed a significantly higher survival rate than those of the other cryoprotectant groups (40% vs 23-26%, p<0.05) and the cleavage rates of PROH, DMSO and EG group reached only 1 to 2%. For a toxicity test of different vitrification media, GV and M II-oocytes were equilibrated stepwise in 100% 8 M EG (group 1) and 1 M SUC + 8 M EG (group 2) or equilibrated in sucrose and then in 8 M EG (SUC+8 M EG, group 3). For GV-oocytes, the survival, maturation and cleavage rates of Group 1 were significantly lower than those in group 2, 3 and control group (p<0.05). For M II-oocytes, there were no differences in survival, maturation and cleavage rates between groups. After vitrification, the survival rates of GV and M II-oocytes in group 2 and 3 were similarly low (4-9%) and none of them matured nor cleaved after in vitro maturation, fertilization and culture. In conclusion, porcine GV and M II-oocytes do not seem to be damaged by a variety of cryoprotectants tested, but will succumb to a temperature decrease to $10^{\circ}C$ or to the process of vitrification, regardless of the cryoprotectant used.

FISH Karyotype Analysis of Seven Rose Cultivars (주요 장미 7품종의 FISH 핵형분석)

  • Hwang, Yoon-Jung;Han, Tae Ho;Martin, Cathie;Lim, Ki-Byung
    • Horticultural Science & Technology
    • /
    • v.30 no.5
    • /
    • pp.568-572
    • /
    • 2012
  • Determination of ploidy level for the mother plant is prerequisite for effective breeding. The study was carried out to determine the ploidy level in 7 different plant materials by FISH karyotype analysis. Among the seven plant varieties analyzed, all exhibit tetraploid (2n = 4x = 28) based on the results observed in chromosome analysis. Four signals of 45S rDNAs were detected on the terminal region of the short arm of chromosome 7. The length of somatic metaphase chromosomes ranges from 1.67 to $2.67{\mu}m$ in 'Alexandra', 1.40 to $2.04{\mu}m$ in 'Freud', 1.64 to 2.24 in 'Little Silver', 1.69 to $2.26{\mu}m$ in 'Teresa', 1.70 to $2.65{\mu}m$ in 'Tineke', 1.35 to $2.08{\mu}m$ in 'Vital', 1.39 to $2.04{\mu}m$ in 'Yellow Mimi'. Total length of the chromosome ranges from $11.23{\mu}m$ in 'Freud' as minimum to $15.05{\mu}m$ in 'Alexandra' as maximum. The karyotypes were composed of metacentric, submetacentric, and subtelocentric chromosome but there is no subtelocentric chromosome.

The Effects of Transcription / Translation Inhibitors on Meiotic Maturation of Porcine Oocyte In Vitro

  • Byun, Tae-Ho;Lee, Sung-Ho;Park, Chang-Sik;Lee, Sang-Ho
    • Proceedings of the Korean Society of Embryo Transfer Conference
    • /
    • 2002.11a
    • /
    • pp.117-117
    • /
    • 2002
  • The oocytes from most of animal species accumulate genetic information and other necessary materials during oogenesis for the later use in the early development. Over the years oocyte maturation has been studied extensively both in vitro and in vivo. Particularly, maturation of follicular oocyte in vitro becomes one of the important tools for the studies of basic cell biology, the in vitro technology of animal production, and in particular, the somatic cell cloning by nuclear transfer. We examined meiotic maturation and cumulus expansion in the presence of translation or transcription inhibitors for varying periods of in viかo maturation (IVM) of pig oocyte. In Experiment 1, the results revealed that translation and transcription inhibitors inhibited cumulus expansion and meiotic maturation during 35h of IVM. However, 50 to 60% of the oocytes underwent nuclear maturation without cumulus expansion during 75h of IVM. The rest of the oocytes were arrested at metaphase I (40-50%) in the presence of the inhibitors. In Experiment II, the OCCs were exposed to the drugs only for 15h to examine translation and transcription inhibitors on cumulus expansion and meiotic maturation. Transcription inhibitors for 15h did not arrest meiotic maturation when the oocytes were cultured for subsequent, necessary period of IVM, whereas cumulus expansion was completely inhibited, suggesting that initial 15h is critical transcription activity far cumulus expansion. Translation inhibitors for 15h exposure did not alter cumulus expansion and meiotic maturation during subsequent culture in the absence of the drugs. In Experiment III, the OCCs were exposed to the drugs only for later 30h to examine the influence of transcription and translation inhibitors on oocyte maturation. Interestingly, all meiotic maturation underwent normally with full expansion of cumulus. Similar results were obtained from Experiment IV where 5h of exposure from 15 to 20h of IVM culture to the drugs was performed and subsequently cultured for same period in fresh medium. Taken there results together, both transcription and translation are necessary for nuclear maturation and cumulus expansion, and first 15h IVM for cumulus expansion is critical. The arrested oocytes by the drugs were still capable of undergoing nuclear maturation, although cumulus expansion was affected.

  • PDF

배양액이 마우스 핵이식 재구축배의 초기발생에 미치는 영향

  • 심보웅;권오용;이은화;김순희;서정선
    • Proceedings of the Korean Society of Embryo Transfer Conference
    • /
    • 2002.11a
    • /
    • pp.76-76
    • /
    • 2002
  • 본 실험은 Piezo-미세조작기(PrimeTech Ltd., Japan)를 사용하여 마우스 핵이식 후 재구축배를 CZB와 KSOM 두가지 배양액을 사용하여 체외배양성적을 비교 검토하였다. MII의 미수정란은 성숙한 4~5주령 B6D2Fl에 hCG 주사 후 14시간째에 과적 방법을 통해 난관의 팽대부로 부터 회수하였고, metaphase II chromosome-spindle complex와 최소량의 세포질을 내경이 10$\mu\textrm{m}$인 피펫으로 흡입하여 탈핵하였다. 핵이식에 사용된 난구세포(8-l0$\mu\textrm{m}$)는 3시간동안 12% PVP 에처리 하여 piezo-미세조작기를 이용하여 세포질에 세포의 핵을 직접 미세주입 하였다. 핵이식 후 생존한 재구축배는 2시간동안 배양한 후 10mM SrC1$_2$와 5$\mu\textrm{g}$/$m\ell$의 cytochalasin B가 첨가된 $Ca^{2+}$-free CZB에서 6시간 활성화 처리하였다, 활성화 처리 후 위전핵이 관찰된 재구축란을 CZB 와 KSOM 배지에서 배양하면서 발달률을 비교하였고, 상실배 및 배반포배로 발달한 재구축배를 day 3 대리모에 이식하였다. 표 1에서 보는 바와 같이 재구축배의 2-cell로의 발달률에 있어서 KSOM이 CZB에 비하여 유의적으로 높게 나타났으며(P<0.05), 또한 4-cell과 상실배/배반 포배로의 발달률에 있어서도 KSOM이 CZB에 비하여 유의적으로 높은 발달률을 나타내었다(P<0.01). 또한 KSOM 배지에서 배양된 상실배/배반포배를 대리모에 이식한 경우에 11.5 d.p.c에 생존한 태아가 관찰되었다. 이상의 결과로 핵이식 재구축배의 활성화 처리 후의 발생에는 KSOM 배지가 CZB 배지에 비하여 유효함을 확인 할 수 있었다.그와 같은 배양 기술을 이용하여 외래유전자를 도입한 일련의 결과에 관하여 보고 하고자한다., 이것은 세포내 유전자가 transfection되지 않은 세포도 neo selection에서 선발된다는 것을 제시하고 있다. 따라서 체세포를 이용한 형질전환동물 생산을 위해서는 세포내 유전자 도입과 선발 과정에서 나타난 colony에 대하여 보다 엄격한 screen을 하는 것이 필요한 것으로 생각된다.로 우점하였다. 여름철 식물플랑크톤 대발생에 영향은 수온과 직산염이 중요하였으나, 부유물질 크게 기여하지 못하였다.애를 확인하고 지도 관점을 파악하는 것을 포함한다. 그러나 본 논문은 역사발생적 수학 학습-지도 원리의 실제적인 적용에 관하여는 기초적인 연구에 지나지 않기 때문에, 역사발생적 원리를 학교수학에 실제적으로 적용하기 위해서는 각각의 내용에 대한 철저한 역사적 분석을 바탕으로 하는 후속 연구가 필요하다./TEX>구성교육${\lrcorner}$이 조선총독부의 관리하에서 실행되었다는 것을, 당시의 사범학교를 중심으로 한 교육조직을 기술한 문헌에 의해 규명시켰다.nd of letter design which represents -natural objects and was popular at the time of Yukjo Dynasty, and there are some documents of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to have been "Japanese Letter Jobcheso." Therefore, the purpose of this study is to look into the origin of the letter designs in t

  • PDF

Recent Advancement on the Knowledges of Meiotic Division (I) (減數分裂, 最近의 進步(I))

  • 한창열
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.6
    • /
    • pp.453-475
    • /
    • 1998
  • During the 100 years since the initial discovery of meiotic phenomenon many brilliant aspects have been elucidated, but further researches based on light microscopy alone as an experimental tool have been found to have some limits and shortcomings. By the use of electron microscopy and armed with the advanced knowledges on modern genetics and biochemistry it has been possible to applu molecular technology in gaining information on the detailed aspects of meiosis. As synapsis takes place, a three-layered proteinous structure called the synatonemal complex starts to form in the space between the homologous chromosomes. To be more precise, it begins to form along the paired chromosomes early in the prophase I of meiotic division. The mechanism that leads to precise point-by-point pairing between homologous chromocomes division. The mechamism that leads to precise point-by-point pairing between homologous chromosomes remains to be ascertained. Several items of information, however, suggest that chromsome alignment leading to synapsis may be mediated somehow by the nuclear membrane. Pachytene bivalents in eukaryotes are firmly attached to the inner niclear membrane at both termini. This attached begins with unpaired leptotene chromosomes that already have developed a lateral element. Once attached, the loptotene chromosomes begin to synapse. A number of different models have been proposed to account for genetic recombination via exchange between DNA strands following their breakage and subsequent reunion in new arrangement. One of the models accounting for molecular recombination leading to chromatid exchange and chiasma formation was first proposed in 1964 by Holliday, and 30 years later still a modified version of his model is favored. Nicks are made by endomuclease at corresponding sites on one strant of each DNA duplex in nonsister chromatid of a bivalent during prophase 1 of meiosis. The nicked strands loop-out and two strands reassociate into an exchanged arrangement, which is sealed by ligase. The remaining intact strand of each duplex is nicked at a site opposite the cross-over, and the exposed ends are digested by exonuclease action. Considerable progress has been made in recent years in the effort to define the molecular and organization features of the centromere region in the yeast chromosome. Centromere core region of the DNA duplex is flanked by 15 densely packed nucleosomes on ons side and by 3 packed nucleosomes on the other side, that is, 2000 bp on one side and 400 400 bp in the other side. All the telomeres of a given species share a common DNA sequence. Two ends of each chromosome are virtually identical. At the end of each chromosome there exist two kinds of DNA sequence" simple telpmeric sequences and telpmere-associated sequencies. Various studies of telomere replication, function, and behabior are now in progress, all greatly aided by molecular methods. During nuclear division in mitosis as well as in meiosis, the nucleili disappear by the time of metaphase and reappear during nuclear reorganizations in telophase. When telophase begins, small nucleoli form at the NOR of each nucleolar-organizing chromosome, enlarge, and fuse to form one or more large nucleoli. Nucleolus is a special structure attached top a specific nucleolar-organizing region located at a specific site of a particular chromosome. The nucleolus is a vertical factory for the synthesis of rRNAs and the assenbly of ribosome subunit precursors.sors.

  • PDF

Effects of Anticancer Agents on Cell Cycle Kinetics and Sister Chromatid Exchanges in Cultured Human Lymphocytes (항암제(抗癌劑)가 배양임파구(培養淋巴球)의 세포분열주기(細胞分裂週期) 및 자매염색분체교환(姉妹染色分體交換)에 미치는 영향(影響))

  • Hwang, In-Dam;Ki, No-Suk;Park, Won-Kihl;Kim, Young-Oh;Lee, Jeong-Sang
    • Journal of Preventive Medicine and Public Health
    • /
    • v.20 no.1 s.21
    • /
    • pp.1-9
    • /
    • 1987
  • Sister chromatid exchanges (SCEs) observed by means of bromodeoxyuridine substitution and fluorescence plus Giemsa (FPG) technique were proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity in short-term cultures of phytohaemagglutinin (PHA)-stimulated human lymphocytes. Therefore, this study was carried out to investigate the relation between anticancer agents and cytotoxic effects. Chromosomal analysis was performed on metaphase cells that had divided one, two, or three or more times after treatment for SCEs, mitotic indices (MI) and cell cycle kinetics by FPG technique. The results indicate that anticancer agents led to a dose dependent increase in SCE frequency except methotrexate. But, highly inhibited mitotic indices and delayed cell cycle kinetics were observed except for cyclophosphamide. The author suggest that the difference of SCE frequency is due to the differences in the cytotoxic action of anticancer agents, but although the induction of SCEs has a correlation with cell cycle delay, in some cases the induction of SCEs is not always related to cell cycle delay because of different cytotoxic action of anticancer agents.

  • PDF

PHA-Induced Peripheral Blood Cytogenetics and Molecular Anslysis : a Valid Diagnostic and Follow-up Modality For Acute Primyelocytic Leukemia Patients Treated With ATRA and/or Arsenic Tri-oxide

  • Baba, Shahid M;Azad, Niyaz A;Shah, Zaffar A;Afroze, Dil;Pandith, Arshad A;Jan, Aleem;Aziz, Sheikh A;Dar, Fayaz A
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.4
    • /
    • pp.1999-2006
    • /
    • 2016
  • Background: Acute promyelocytic leukemia (APML) is characterized by the reciprocal translocation t(15;17) (p22;p12) resulting in the PML-$RAR{\alpha}$ fusion gene. A dual diagnostic and follow up approach was applied including cytogenetic demonstration of the t(15;17) translocation and detection dg PML-$RAR{\alpha}$ chimeric transcripts by molecular means. Purpose: Conventional cytogenetics involving bone marrow is beset with high probability of poor metaphase index and was substituted with phytohemagglutinin (PHA)-induced peripheral blood culture based cytogenetic analysis as a diagnostic & follow up modality in APML patients of Kashmir (North India). Both qualitative (RT-PCR) and quantitative (Q-PCR) tests were simultaneously carried out to authenticte the modified cytogenetics. Materials and Method: Patient samples were subjected to the said techniques to establish their baseline as well as follow-up status. Results: Initial cytogenetics revealed 30 patients (81%) Positive for t(15;17) whereas 7 (19%) had either cryptic translocation or were negative for t(15;17). Two cases had chromosome 16q deletion and no hallmark translocation t(15;17). Q-PCR status for PML-$RAR{\alpha}$ was found to be positive for all patients. All the APML patients were reassessed at the end of consolidation phase and during maintenance phase of chemotherapy where 6 patients had molecular relapse, wherein 4 also demonstrated cytogenetic relapse. Conclusions: It was found that PHA-induced peripheral blood cytogenetics along with molecular analysis could prove a reliable modality in the diagnosis and assessment of follow up response of APML patients.