• Title/Summary/Keyword: Experimental Kit

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Differences in 25-hydroxy vitamin D and vitamin D-binding protein concentrations according to the severity of endometriosis

  • Baek, Jong Chul;Jo, Jae Yoon;Lee, Seon Mi;Cho, In Ae;Shin, Jeong Kyu;Lee, Soon Ae;Lee, Jong Hak;Cho, Min-Chul;Choi, Won Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.3
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    • pp.125-131
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    • 2019
  • Objective: To investigate serum 25-hydroxyl vitamin D (25(OH)D) and vitamin D-binding protein (VDBP) concentrations in women with endometriosis according to the severity of disease. Methods: Women with mild endometriosis (n = 9) and advanced endometriosis (n = 7), as well as healthy controls (n = 16), were enrolled in this observational study. Serum total 25(OH)D concentrations were analyzed using the Elecsys vitamin D total kit with the Cobas e602 module. Concentrations of bioavailable and free 25(OH)D were calculated. Concentrations of VDBP were measured using the Human Vitamin D BP Quantikine ELISA kit. Variables were tested for normality and homoscedasticity using the Shapiro-Wilk test and Leven F test, respectively. Correlation analysis was used to identify the variables related to total 25(OH)D and VDBP levels. To assess the effects of total 25(OH)D and VDBP levels in the three groups, multivariate generalized additive modeling (GAM) was performed. Results: Gravidity and parity were significantly different across the three groups. Erythrocyte sedimentation rate (ESR) and CA-125 levels increased as a function of endometriosis severity, respectively (p= 0.051, p= 0.004). The correlation analysis showed that total 25(OH)D levels were positively correlated with gravidity (r = 0.59, p< 0.001) and parity (r = 0.51, p< 0.003). Multivariate GAM showed no significant relationship of total 25(OH)D levels with EMT severity after adjusting for gravidity and ESR. However, the coefficient of total 25(OH)D levels with gravidity was significant (1.87; 95% confidence interval, 0.12-3.63; p= 0.040). Conclusion: These results indicate that vitamin D and VDBP levels were not associated with the severity of endometriosis.

Sperm chromatin structure assay versus sperm chromatin dispersion kits: Technical repeatability and choice of assisted reproductive technology procedure

  • Laxme B, Vidya;Stephen, Silviya;Devaraj, Ramyashree;Mithraprabhu, Sridurga;Bertolla, Ricardo P.;Mahendran, Tara
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.4
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    • pp.277-283
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    • 2020
  • Objective: The sperm DNA fragmentation index (DFI) guides the clinician's choice of an appropriate assisted reproductive technology (ART) procedure. The DFI can be determined using commercially available methodologies, including sperm chromatin dispersion (SCD) kits and sperm chromatin structure assay (SCSA). Currently, when DFI is evaluated using SCD kits, the result is analyzed in reference to the SCSA-derived threshold for the choice of an ART procedure. In this study, we compared DFI values obtained using SCSA with those obtained using SCD and determined whether the difference affects the choice of ART procedure. Methods: We compared SCSA to two SCD kits, CANfrag (n=36) and Halosperm (n=31), to assess the DFI values obtained, the correlations between tests, the technical repeatability, and the impact of DFI on the choice of ART. Results: We obtained higher median DFI values using SCD kits than when using SCSA, and this difference was significant for the CANfrag kit (p<0.001). The SCD kits had significantly higher coefficients of variation than SCSA (p<0.001). In vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) would be chosen for a significantly higher proportion of patients if a decision were made based on DFI derived from SCD rather than DFI determined using SCSA (p=0.003). Conclusion: Our results indicate that SCD kit-specific thresholds should be established in order to avoid the unnecessary use of IVF/ICSI based on sperm DNA damage for the management of infertility. Appropriate measures should be taken to mitigate the increased variability inherent to the methods used in these tests.

Experimental Studies on the Hair Growth Activity of Fractions and Extract of Arisaematis Rhizoma in C57B/6N Mice (C57BL/6N 생쥐에서 천남성 추출물과 분획물의 발모효과에 대한 실험적 연구)

  • Kwon, Kyung-Suk;Lee, Moon-Won;Jeong, Il-Kook;Jeong, Han-So;Song, Beom-Yong;Song, Jeong-Mo;Lee, Chang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.619-630
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    • 2009
  • To investigate the hair growth activity of fractions and extract of Arisaematis Rhizoma in the hair removed skin of normal and spontaneous alopecia areata model in C57B/6N mice. These experiments were performed with the macroscopic, microscopic, immunohistochemical(VEGF, c-kit, PKC-${\alpha}$, TGF and FGF) and RT-PCR(TGF-${\beta}$, IGF, prolactin and placenta lactogen) methods. The results were as follows: Macroscopic observation after topical application of vehicle, 50% EtOH as control and extract of Arisaematis Rhizoma to the hair removed skin of C57BL/6N mice on the 9th, 11th and 15th day. Extensive hair growth activity was observed in treated group with extract of Arisaematis Rhizoma on the 9th, 11th and 15th day. In Arisaematis Rhizoma extracts treated group, hair follicles of middle stage of anagen was observed and it were grown down to subcutaneous tissue of skin in all the normal mice on 15th day. But in control group, most of hair follicles of telogen phase was observed in skin. The treatment of extract of Arisaematis Rhizoma increased expression of IGF(145%) and placenta lactogen(108%) in the skin of normal C57BL/6N mice on the 11th day compared to control group(100%). But expression of TGF-${\beta}$(90%) and prolactin(91%) decreased in the skin of normal C57B/6N mice on the 11th day compared to control group(100%). After application of fractions(chloroform, ethyl acetate and water fractions) of Arisaematis Rhizoma extract for 9th day, hair growth effect was observed in whole skin area in 50% of normal mice. But in control group, hair growth effect was not observed in whole skin area of normal mice. Immunoreactive density of VEGF, c-kit, PKC-${\alpha$ and FGF in skin of fractions of Arisaematis Rhizoma extracts was strongly stained in epidermis, bulge, secondary hair germ cells, cutaneous trunci m., subcutaneous tissue, root sheath compare to control group on the 9th day. In spontaneous alopecia areata model, The hair growth activity of Arisaematis Rhizoma extrat treated group(75%) was observed to be strong compared to control group(O%) on 7th day. These experiments suggest that fractions and extracts of Arisaematis Rhizoma may stimulate the topical hair growth activity. Thus it can be useful for treatment of alopecia areata.

mRNA Expression of the Regulatory Factors for the Early Folliculogenesis in vitro (체외배양 중인 생쥐 난소에서 초기난포 조절인자의 발현)

  • Yoon, Se-Jin;Kim, Ki-Ryeong;Chung, Hyung-Min;Yoon, Tae-Ki;Cha, Kwang-Yul;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.3
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    • pp.207-216
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    • 2005
  • Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

Strengthening of non-seismically designed beam-column joints by ferrocement jackets with chamfers

  • Li, Bo;Lam, Eddie Siu-Shu;Cheng, Yuk-Kit;Wu, Bo;Wang, Ya-Yong
    • Earthquakes and Structures
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    • v.8 no.5
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    • pp.1017-1038
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    • 2015
  • This paper presents a strengthening method that involves the use of ferrocement jackets and chamfers to relocate plastic hinge for non-seismically designed reinforced concrete exterior beam-column joints. An experimental study was conducted to assess the effectiveness of the proposed strengthening method. Four half-scale beam-column joints, including one control specimen and three strengthened specimens, were prepared and tested under quasi-static cyclic loading. Strengthening schemes include ferrocement jackets with or without skeleton reinforcements and one or two chamfers. Experimental results have indicated that the proposed strengthening method is effective to move plastic hinge from the joint to the beam and enhance seismic performance of beam-column joints. Shear stress and distortion within the joint region are also reduced significantly in strengthened specimens. Skeleton reinforcements in ferrocement provide limited improvement, except on crack control. Specimen strengthened by ferrocement jackets with one chamfer exhibits slight decrease in peak strength and energy dissipation but with increase in ductility as compared with that of two chamfers. Finally, a method for estimating moment capacity at beam-column interface for strengthened specimen is developed. The proposed method gives reasonable prediction and can ensure formation of plastic hinge at predetermined location in the beam.

Curcumin Induces Caspase Mediated Apoptosis in JURKAT Cells by Disrupting the Redox Balance

  • Gopal, Priya Kalyan;Paul, Mausumi;Paul, Santanu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.93-100
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    • 2014
  • Background: Curcumin has has been reported to exert anti-inflammatory, anti-oxidation and anti-angiogenic activity in various types of cancer. It has also been shown to induce apoptosis in leukemia cells. We aimed to unravel the role of the redox pathway in Curcumin mediated apoptosis with a panel of human leukemic cells. Materials and Methods: In this study in vitro cytotoxicity of Curcumin was measured by MTT assay and apoptotic effects were assessed by annexin V/PI, DAPI staining, cell cycle analysis, measurement of caspase activity and PARP cleavage. Effects of Curcumin on intracellular redox balance were assessed using fluorescent probes like $H_2DCFDA$, JC1 and an ApoGSH Glutathione Detection Kit respectively. Results: Curcumin showed differential anti-proliferative and apoptotic effects on different human leukemic cell lines in contrast to minimal effects on normal cells. Curcumin induced apoptosis was associated with the generation of intracellular ROS, loss of mitochondrial membrane potential, intracellular GSH depletion, caspase activation. Conclusions: As Curcumin induces programmed cell death specifically in leukemic cells it holds a great promise as a future therapeutic agent in the treatment of leukemia.

Characterization of the Salmonella typhi Outer Membrane Protein C

  • Toobak, Hoda;Rasooli, Iraj;Gargari, Seyed Latif Mousavi;Jahangiri, Abolfazl;Nadoushan, Mohammadreza Jalali;Owlia, Parviz;Astaneh, Shakiba Darvish Alipour
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.128-134
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    • 2013
  • Salmonella enterica serovar typhi, a Gram-negative food-borne pathogen, causes typhoid fever in humans. OmpC is an outer membrane porin of S. typhi expressed throughout the infection period. OmpC is potentially an attractive antigen for multivalent vaccines and diagnostic kit designs. In this study we combined in silico, in vitro and in vivo approaches to analyze various aspects of OmpC's antigenic properties. The conserved region, in addition to secondary and tertiary structures, and linear B cell epitopes, were predicted. A number of results obtained from in silico analyses were validated by experimental studies. OmpC was amplified, cloned and then expressed, with the recombinant protein then being purified. BALB/c mice were immunized by purified denatured OmpC. The titer of antibody was raised. Results of challenges with the pathogen revealed that the immunity is non-protective. Most of the theoretical and experimental results were in consensus. Introduced linear B cell epitopes can be employed for the design of diagnostic kits based on antigen-antibody interactions.

Large eddy simulation of turbulent flow using the parallel computational fluid dynamics code GASFLOW-MPI

  • Zhang, Han;Li, Yabing;Xiao, Jianjun;Jordan, Thomas
    • Nuclear Engineering and Technology
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    • v.49 no.6
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    • pp.1310-1317
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    • 2017
  • GASFLOW-MPI is a widely used scalable computational fluid dynamics numerical tool to simulate the fluid turbulence behavior, combustion dynamics, and other related thermal-hydraulic phenomena in nuclear power plant containment. An efficient scalable linear solver for the large-scale pressure equation is one of the key issues to ensure the computational efficiency of GASFLOW-MPI. Several advanced Krylov subspace methods and scalable preconditioning methods are compared and analyzed to improve the computational performance. With the help of the powerful computational capability, the large eddy simulation turbulent model is used to resolve more detailed turbulent behaviors. A backward-facing step flow is performed to study the free shear layer, the recirculation region, and the boundary layer, which is widespread in many scientific and engineering applications. Numerical results are compared with the experimental data in the literature and the direct numerical simulation results by GASFLOW-MPI. Both time-averaged velocity profile and turbulent intensity are well consistent with the experimental data and direct numerical simulation result. Furthermore, the frequency spectrum is presented and a -5/3 energy decay is observed for a wide range of frequencies, satisfying the turbulent energy spectrum theory. Parallel scaling tests are also implemented on the KIT/IKET cluster and a linear scaling is realized for GASFLOW-MPI.

Development of Chemiluminescence Immunoassay for Progesterone in Serum (혈청내의 Progesterone 측정을 위한 Chemiluminescence Immunoassay의 개발에 관한 연구)

  • Lee, K.S.;Suh, B.H.;Lee, J.H.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.87-91
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    • 1990
  • Development of a solid-phase chemiluminescence immunoassay for the detection of progesterone in serum extract was described. The chemiluminescence immunoassay was establised utilizing anti-progesterone monoclonal antibody that coated on polystyrene tubes and progesterone-ABEI conjugate as tracer. The light yield generated from antibody bound conjugate was counted on clinilumat luminometer by oxidation with microperoxidase and peroxide. The chemiluminescence immunoassay was high specific and accurate and detects as little as 3.9ng/ml of progesterone. The intra-assay CV ranged from 6% to 11.5% and inter-assay CV ranged from 13.6% to 18.7%. This assay system was good correlated with conventional kit radioimmunoassay system (r=0.98).

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Mutagenecity Test of SDK (SDK시제품(가칭)에 대한 변이원성시험)

  • 정지윤;이원우;임종희;남정석;제정환;이광훈;강병철;이병희;박재학
    • Toxicological Research
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    • v.14 no.2
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    • pp.211-216
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    • 1998
  • In order to evaluate the mutagenic potential of SDK(skin decontamination kit) produced by Agency for Defense Development(ADD), were performed Salmonella typhimurium reversion assay, chromosomal aberration test on chinese hamster ovarian cells and in vivo micronucleus assay using mouse bone marrow cells according to the established regulation of Korean Food and Drug Administration. In the reverse mutation test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 did not in-crease the number of revertant at any of the concentration tested in this study. SDK did not increase the number of cells having structural or numerical chromosome aberration in cytogenetic test. In mouse micronucleus test, no significant increase in the occurrence oj micro nucleated polychromatic erythrocytes were observed in ICR male mice intraperitoneally administered with SDK. These results indicate that SDK has no mutagenic effects under these experimental conditions.

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