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A Comparative Analysis of the Illumina Truseq Synthetic Long-read Haplotyping Sequencing Platform versus the 10X Genomics Chromium Genome Sequencing Platform for Haplotype Phasing and the Identification of Single-nucleotide variants (SNVs) in Hanwoo (Korean Native Cattle) (일루미나에서 제작된 TSLRH (Truseq Synthetic Long-Read Haplotyping)와 10X Genomics에서 제작된 The Chromium Genome 시퀀싱 플랫폼을 이용하여 생산된 한우(한국 재래 소)의 반수체형 페이징 및 단일염기서열변이 비교 분석)

  • Park, Woncheoul;Srikanth, Krishnamoorthy;Park, Jong-Eun;Shin, Donghyun;Ko, Haesu;Lim, Dajeong;Cho, In-Cheol
    • Journal of Life Science
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    • v.29 no.1
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    • pp.1-8
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    • 2019
  • In Hanwoo cattle (Korean native cattle), there is a scarcity of comparative analysis papers using highdepth sequencing and haplotype phasing, particularly a comparative analysis of the Truseq Synthetic Long-Read Haplotyping sequencing platform serviced by Illumina (TSLRH) versus the Chromium Genome Sequencing platform serviced by 10X Genomics (10XG). DNA was extracted from the sperm of a Hanwoo breeding bull (ID: TN1505D2184/27214) provided by Hanwoo research canter and used for the generation of sequence data from both the sequencing platforms. We then identified SNVs using an appropriate analysis pipeline tailored for each platform. The TSLRH and 10XG platforms generated a total of 355,208,304 and 1,632,772,004 reads, respectively, corresponding to a Q30 (%) of 89.04% and 88.60%, respectively, of which 351,992,768(99.09%) and 1,526,641,824(93.50%) were successfully mapped. For the TSLRH and 10XG platforms, the mean depth of the sequencing was 13.04X and 74.3X, the longest phase block was 1,982,706 bp and 1,480,081 bp, the N50 phase block was 57,637 bp and 114,394 bp, the total number of SNVs identified was 4,534,989 and 8,496,813, and the total phased rate was 72.29% and 87.67%, respectively. Moreover, for each chromosome, we identified unique and common SNVs using both sequencing platforms. The number of SNVs was directly proportional to the length of the chromosome. Based on our results, we recommend the use of the 10XG platform for haplotype phasing and SNV identification, as it generated a longer N50 phase block, in addition to a higher mean depth, total number of reads, total number of SNVs, and phase rate, than the TSLRH platform.

Improvement in Calculating Engineer Standard Wage Rate and Its Appropriate Level Computation (엔지니어링 노임단가 산출기준 개선방안과 적정 노임단가 추정)

  • Lee, Jae Yul;Lee, Hae Kyung
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.42 no.6
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    • pp.853-860
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    • 2022
  • The purpose of this study is to suggest an improvement plan for the calculation method of the engineer standard wage rate (ESWR) and to compute a reasonable ESWR. To this end, an adequacy review of theESWR calculation criteria was conducted along with an extensive engineering industry survey. The survey results were analyzed using an effective response sample of 748 companies out of 1,000 survey samples extracted by stratifying the 5,879 survey population. The main results were as follows. ①When calculating the engineering service fee, the prime contractor's engineer wage is suitable for the ESWR. The ESWR can be estimated by the formula 'average wage÷[1-proportion of subcontract orders×(1-subcontract rate)].' ② The field survey showed that the number of monthly working days was 20.35-20.54 days at 99 % confidence interval, which was significantly different from the current standard (22 days). In addition, as a result of a legal review of the ESWR criteria, it was found that the number of working days should be calculated in accordance with the Labor Standards Act after 2022. ③ Applying government guidelines, the time difference between the wage survey and the ESWR application can be corrected by the past ESWR increase rate for a specific period. ④ Using modeling based on the analysis above, the current ESWR was 13.5-14.5 % lower than the appropriate level. A lower ESWR was driven by the non-reflection of subcontract structure (4.1 %), overestimation of monthly work days (6.8-7.8 %), and application of past wage (2.6 %). The proposed model is expected to be widely used in policy making, as it can provide a useful framework for calculating the standard wage rate in similar industries as well as calculating appropriate engineering fees.

Effect of Sperm Preincubation Medium with Ascorbic Acid and/or Ferrous Sulfate on Porcine In-Vitro Fertilization (돼지의 체외수정시 Ascorbic Acid와 Ferrous Sulfate의 첨가하에서 정자 전배양의 영향)

  • Park, C.K.;Nam, H.S.;Lee, J.H.;Kim, I.C.;Cheong, H.T.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.255-262
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    • 2000
  • The aim of this work was to study the effects of ascorbic acid (Asc) and/or ferrous sulfate (Fe$^{2+}$) and spernatozoa preincubation on the in vitro fertilization in porcine. Porcine follicular oocytes matured in culture were inseminated with frozen-thawed boar spermatozoa preincubated for 0, 1, 2, 3, 4 and 5h. The penetration rates (37~51%) were not significantly different between durations of spermatozoa preincubation in medium with 0.1 mM Asc. The addition of 1.0 mM Fe$^{2+}$ during spermatozoa preincubation were not significantly affecting the penetration rates (41~56%). When spermatozoa were preincubated with Asc and Fe$^{2+}$, the penetration rates had a tendency to increase with time of spermatozoa preincubation, and were significantly (P<0.05) higher in spermatozoa preincubated with that than without Asc and Fe$^{2+}$ for 5 h. On the other hand, when spermatozoa were preincubated in fertilization medium without Asc and/or Fe$^{2+}$, the penetration rates were significantly (P<0.05) higher in medium with Fe$^{2+}$ than with Asc or Asc and Fe$^{2+}$ for in vitro fertilization. The rate of polyspermy in penetrated oocytes in medium with Asc and Fe$^{2+}$ decreased with the period of spermatozoa preincubation. Despite different culture conditions for spermatozoa preincubation, no differences were observed in polyspermy rates in the presence of Asc and/or Fe$^{2+}$ These results indicate the advantage of preincubating spermatozoa with Asc and Fe$^{2+}$ and an addition of Fe$^{2+}$ during in vitro fertilization with spermatozoa preincubated maintain penetration potential without increased polyspermy rates on in vitro fertilization in porcine oocytes.on in porcine oocytes.

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Studies on Genetics and Breeding in Rainbow Trout(Oncorhynchus mykiss) VII. Fertilization of Fresh Egg with Co-Preserved Sperm and Ultrastructural Changes (무지개 송어의 유전 육종학적 연구 VII. 동결보존시킨 정자와 신선한 난모세포의 수정 및 미세구조적 변화)

  • PARK Hong-Yang;YOON Jong-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.79-92
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    • 1992
  • This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

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Review of Sexual Dysfunction in Male Schizophrenics (남자 정신분열병 환자에서 성기능장애에 대한 검토)

  • Choi, Yeong Tae;Cheon, Jin Sook;Oh, Byoung Hoon
    • Korean Journal of Biological Psychiatry
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    • v.7 no.1
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    • pp.85-98
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    • 2000
  • Objective : There are four possible explanations for the sexual dysfunction of schizophrenics. The first is the possibility of a real structural aspect. The second possibility is that sexual function changes secondary to the illness. The third possibility is that there are medical and sociocultural barriers to sexual expression for chronic schizophrenics. The fourth possibility is that sexual dysfunction due to antipsychotic medication. However, we didn't know the precise cause of sexual dysfunction in schizophrenics. Therefore, the purpose of this study was to explore the mechanism of illness itself and antipsychotics on sexual dysfunction in male schizophrenics. Methods : The serum prolactin(PRL), testosterone(TST), and the plasma serotonin(5-HT) concentrations were measured by radioimmunoassay and high performance liquid chromatography method for 100 healthy male schizophrenics according to the DSM-IV. Concomitantly, the severity of psychotic symptoms using Clinical Global Impression(CGI), Brief Psychiatric Rating Scale(BPRS), Positive and Negative Syndrome Scale(PANSS), and the severity of side effects for antipsychotics using Extrapyramidal Side Effects Scale(EPSE), Anticholinergic Side Effects Scale(ACSE), the cognitive function using PANSS-Cognitive Function(PANSS-CF), Mini Mental State Exam-Korean(MMSE-K), and the sexual dysfunction using Sexual Functioning Questionnaire(SFQ), Questionnaire for Sexual Dysfunction in Men were assessed. The PRL, TST, and 5-HT levels of 50 healthy male controls who had no medical, neurological, and psychiatric illnesses were evaluated. The sexual function using SFQ(items FGa, FNa) were also assessed. Furthermore, the correlation with age, education, religion, economic status, age at onset, duration of illnesses, duration of admission, levels of PRL, TST, 5-HT, antipsychotic dosages, potency, benztropine, total duration of medication, EPSE, ACSE, CGI, BPRS, PANSS, PANSS-CF, MMSE-K and sexual dysfunctions were identified in male schizophrenics. Results : 1) The frequencies of sexual dysfunctions for schizophrenics(80%) were significantly(p<0.001) higher than those for controls(42%). The sexual dysfunctions according to sexual response cycle were 'low sexual desire' 76%, 'impairment of achieving erection' 75%, 'impairment of maintaining erection' 75%, 'impairment of obtaining orgasm' 32%, 'impairment in the quality of orgasm' 61%, 'impairment in quantity of ejaculate' 44%, 'premature ejaculation' 15%, and 'delayed ejaculation' 50%. 2) The PRL, 5-HT levels of schizophrenics($28.5{\pm}20.6ng/ml$, $298.5{\pm}89.1ng/ml$) were significantly(p<0.001) higher than those of controls($10{\pm}5.6ng/ml$, $169.2{\pm}37.8ng/ml$), while the TST levels of schizophrenics($4.3{\pm}1.5ng/ml$) and controls($4.5{\pm}1.2ng/ml$) were not significantly different. The sexual dysfunctions of schizophrenics who had abnormal 5-HT levels($4.7{\pm}1.3$ scores) were significantly(p<0.05) higher than those of who had normal 5-HT levels($3.8{\pm}1.6$ scores) on item D7. 3) The sexual dysfunctions of unmarried schizophrenics were significantly(p<0.01 : p<0.05) higher than those of married schizophrenics($6.1{\pm}2.8$ scores, $4.7{\pm}1.3$ scores on item FGa : ${\beta}$=-0.211 on item FNa). The sexual dysfunctions were positively correlated with the rise of 5-HT levels(r=0.209, p<0.05 on item D4 and r=0.241, p<0.05 on item D7), the higher age at onset(r=0.275, p<0.01 on item FNa : r=-0.202, p<0.05 on item FDa), the longer duration of illnesses(r=0.237, p<0.05 on item D6), the longer duration of admission(r=0.234, p<0.05 on item D4 : r=0.328, p<0.05 on item D6), the longer total duration of medication(r=0.237, p<0.05 on item D6). However, age, education, religion, economic status, PRL, TST levels, antipsychotics dosage, potency, benztropine, ACSE, CGI, BPRS, PANSS, PANSS-CF, MMSE-K scores were not correlated with increased sexual dysfunctions. Conclusions : Male schizophrenics have significantly more sexual dysfunction to compare with controls. The higher frequencies of sexual dysfunctions were low sexual desire and erectile disorder. The unmarried, higher age at onset, and longer duration of diseases were positively correlated with increased sexual dysfunctions. Also high 5-HT levels were positively correlated with increased sexual dysfunctions. This means that studies of plasma 5-HT levels, albeit questionable indicators of central 5-HT function, offer some additional support for the association of sexual dysfunction with excess 5-HT activity as primary pathology of schizophrenia. Our findings suggest that excess 5-HT activity seems to affect the patient's sexual function.

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