• Title/Summary/Keyword: live cell

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Differential Parental Transmission of Markers in BCL3 among Korean Cleft Case-parent Trios

  • Park, Beyoung-Yun;Sull, Jae-Woong;Park, Jung-Yong;Jee, Sun-Ha;Beaty, Terri H.
    • Journal of Preventive Medicine and Public Health
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    • v.42 no.1
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    • pp.1-4
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    • 2009
  • Objectives : Isolated cleft lip with or without cleft palate(CL/P) is among the most common human birth defects, with a prevalence of approximately 1 in 700 live births. The B-Cell Leukemia/lymphoma 3(BCL3) gene has been suggested as a candidate gene for CL/P based on association and linkage studies in some populations. This study tests for an association between markers in BCL3 and isolated, non-syndromic CL/P using a case-parent trio design, while considering parent-of-origin effects. Methods : Forty case-parent trios were genotyped for two single nucleotide polymorphisms(SNPs) in the BCL3 gene. We performed a transmission disequilibrium test(TDT) on individual SNPs, and the FAMHAP package was used to estimate haplotype frequencies and to test for excess transmission of multi-SNP haplotypes. Results : The odds ratio for transmission of the minor allele, OR(transmission), was significant for SNP rs8100239(OR=3.50, p=0.004) and rs2965169(OR=2.08, p=0.027) when parent-of-origin was not considered. Parentspecific TDT revealed that SNP rs8100239 showed excess maternal transmission. Analysis of haplotypes of rs2965169 and rs8100239 also suggested excess maternal transmission. Conclusions : BCL3 appears to influence risk of CL/P through a parent-of-origin effect with excess maternal transmission.

Prevalence of autoantibodies that bind to kidney tissues in cats and association risk with antibodies to feline viral rhinotracheitis, calicivirus, and panleukopenia

  • Songaksorn, Nisakorn;Petsophonsakul, Wilaiwan;Pringproa, Kidsadagon;Lampang, Kannika Na;Sthitmatee, Nattawooti;Srifawattana, Nuttawan;Piyarungsri, Kakanang;Thongkorn, Kriangkrai
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.38.1-38.17
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    • 2021
  • Background: The feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine, prepared from viruses grown in the Crandell-Rees feline kidney cell line, can induce antibodies to cross-react with feline kidney tissues. Objectives: This study surveyed the prevalence of autoantibodies to feline kidney tissues and their association with the frequency of FVRCP vaccination. Methods: Serum samples and kidneys were collected from 156 live and 26 cadaveric cats. Antibodies that bind to kidney tissues and antibodies to the FVRCP antigen were determined by enzyme-linked immunosorbent assay (ELISA), and kidney-bound antibody patterns were investigated by examining immunofluorescence. Proteins recognized by antibodies were identified by Western blot analysis. Results: The prevalences of autoantibodies that bind to kidney tissues in cats were 41% and 13% by ELISA and immunofluorescence, respectively. Kidney-bound antibodies were observed at interstitial cells, apical border, and cytoplasm of proximal and distal tubules; the antibodies were bound to proteins with molecular weights of 40, 47, 38, and 20 kDa. There was no direct link between vaccination and anti-kidney antibodies, but positive antibodies to kidney tissues were significantly associated with the anti-FVRCP antibody. The odds ratio or association in finding the autoantibody in cats with the antibody to FVRCP was 2.8 times higher than that in cats without the antibody to FVRCP. Conclusions: These preliminary results demonstrate an association between anti-FVRCP and anti-cat kidney tissues. However, an increase in the risk of inducing kidney-bound antibodies by repeat vaccinations could not be shown directly. It will be interesting to expand the sample size and follow-up on whether these autoantibodies can lead to kidney function impairment.

Effects of winter and spring housing on growth performance and blood metabolites of Pengbo semi-wool sheep in Tibet

  • Jin, Yan Mei;Zhang, Xiao Qing;Badgery, Warwick B;Li, Peng;Wu, Jun Xi
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1630-1639
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    • 2019
  • Objective: Sixty Pengbo semi-wool sheep ewes (approximately 1.5-years-old; $31.33{\pm}0.43kg$) were randomly assigned to two groups, either grazing (G) or dry lot feeding (D), to examine the effects of traditional daily grazing and dry lot feeding on performance and blood metabolites during the cold season in Tibetan Plateau. Methods: The ewes in the G group were grazed continuously each day and housed in one shed each evening, while the ewes in the D group were housed in another shed all day. All animals were fed 400 g/d of commercial concentrate, and grass hay was available freely throughout the experimental period. Results: Compared with the G group, the ewes in the D group had higher (p<0.05) live weight and weight gain. The D group ewes had greater (p<0.05) numbers of white blood cells and platelets, while they had lower (p<0.05) platelet-large cell ratios, cholesterol, high-density lipoprotein cholesterol and glutathione peroxidase, as compared with the G group ewes. Additionally, three serum metabolites, abscisic acid, xanthoxin and 3,4-dihydroxy-5-polypren, were upregulated (p<0.05) in the G group in comparison with the D group. Conclusion: In conclusion, a dry lot feeding regime during the winter and spring period will increase the productivity of sheep and improve blood physiological and biochemical profiles.

In-vitro meat: a promising solution for sustainability of meat sector

  • Kumar, Pavan;Sharma, Neelesh;Sharma, Shubham;Mehta, Nitin;Verma, Akhilesh Kumar;Chemmalar, S;Sazili, Awis Qurni
    • Journal of Animal Science and Technology
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    • v.63 no.4
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    • pp.693-724
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    • 2021
  • The in-vitro meat is a novel concept in food biotechnology comprising field of tissue engineering and cellular agriculture. It involves production of edible biomass by in-vitro culture of stem cells harvested from the muscle of live animals by self-organizing or scaffolding methodology. It is considered as efficient, environmental friendly, better ensuring public safety and nutritional security, as well as ethical way of producing meat. Source of stem cells, media ingredients, supply of large size bioreactors, skilled manpower, sanitary requirements, production of products with similar sensory and textural attributes as of conventional meat, consumer acceptance, and proper set up of regulatory framework are challenges faced in commercialization and consumer acceptance of in-vitro meat. To realize any perceivable change in various socio-economic and environmental spheres, the technology should be commercialized and should be cost-effective as conventional meat and widely accepted among consumers. The new challenges of increasing demand of meat with the increasing population could be fulfill by the establishment of in-vitro meat production at large scale and its popularization. The adoption of in-vitro meat production at an industrial scale will lead to self-sufficiency in the developed world.

Magnetic-activated cell sorting improves high-quality spermatozoa in bovine semen

  • de Assumpcao, Teresinha Ines;Severo, Neimar Correa;Zandonaide, Joao Pedro Brandao;Macedo, Gustavo Guerino
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.2
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    • pp.91-98
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    • 2021
  • The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 106 cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 106 cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.

Methanol extracts of Humulus japonicus induced apoptosis in human FaDu hypopharynx squamous carcinoma cells

  • Jang, Ji Yeon;Park, Bo-Ram;Lee, Seul Ah;Choi, Mi Suk;Kim, Chun Sung
    • International Journal of Oral Biology
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    • v.47 no.1
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    • pp.9-15
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    • 2022
  • Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.

Proliferation of Mouse Prostate Cancer Cells Inflamed by Trichomonas vaginalis

  • Kim, Sang-Su;Kim, Kyu-Shik;Han, Ik-Hwan;Kim, Yeseul;Bang, Seong Sik;Kim, Jung-Hyun;Kim, Yong-Suk;Choi, Soo-Yeon;Ryu, Jae-Sook
    • Parasites, Hosts and Diseases
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    • v.59 no.6
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    • pp.547-556
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    • 2021
  • Our objective was to investigate whether inflammatory microenvironment induced by Trichomonas vaginalis infection can stimulate proliferation of prostate cancer (PCa) cells in vitro and in vivo mouse experiments. The production of CXCL1 and CCL2 increased when cells of the mouse PCa cells (TRAMP-C2 cell line) were infected with live T. vaginalis. T. vaginalis-conditioned medium (TCM) prepared from co-culture of PCa cells and T. vaginalis increased PCa cells migration, proliferation and invasion. The cytokine receptors (CXCR2, CCR2, gp130) were expressed higher on the PCa cells treated with TCM. Pretreatment of PCa cells with antibodies to these cytokine receptors significantly reduced the proliferation, mobility and invasiveness of PCa cells, indicating that TCM has its effect through cytokine-cytokine receptor signaling. In C57BL/6 mice, the prostates injected with T. vaginalis mixed PCa cells were larger than those injected with PCa cells alone after 4 weeks. Expression of epithelial-mesenchymal transition markers and cyclin D1 in the prostate tissue injected with T. vaginalis mixed PCa cells increased than those of PCa cells alone. Collectively, it was suggested that inflammatory reactions by T. vaginalis-stimulated PCa cells increase the proliferation and invasion of PCa cells through cytokine-cytokine receptor signaling pathways.

Apoptosis induced by water extracts of Nypa fruticans wurmb via a mitochondria-dependent pathway in human FaDu hypopharyngeal squamous carcinoma cells

  • Lee, Seul Ah;Choi, Mi Suk;Park, Bo-Ram;Kim, Jin-Soo;Kim, Chun Sung
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.160-167
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    • 2021
  • Nypa fruticans Wurmb (NFW) contains a large amount of phenolic acid and flavonoids, and is popular as a superfood in Myanmar. NFW has various biological activities, such as anti-inflammatory, anti-oxidant, and neuroprotective properties; however, the anti-cancer effect of NFW have not been reported. In this study, we investigated the anticancer activity of water extracts of NFW (WeNFW) and the underlying mechanism in human FaDu hypopharyngeal squamous carcinoma cells. The WeNFW inhibited FaDu cell growth in a dose-dependent manner without affecting normal cells (L929), as determined by an MTT assay and Live and Dead assay. In addition, the concentrations of WeNFW without cytotoxicity (0.025, 0.05, and 0.1 mg/mL) inhibited wound healing and colony formation. Furthermore, WeNFW significantly induced apoptosis through the proteolytic cleavage of caspase-3 and -9, poly (ADP-ribose) polymerase, and downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by DAPI staining, FACS analysis, and western blot analysis. Taken together, these results suggest that WeNFW exhibits potent anti-cancer effects by suppressing the growth of oral cancer cells, wound healing and colony formation activity. Via mitrochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, WeNFW can provide a natural chemotherapeutic drug for oral cancer in humans.

Limonium Tetragonum Enhances Osteoblastogenesis while Suppressing the Adipocyte Differentiation

  • Kim, Jung-Ae;Ahn, Byul-Nim;Oh, Jung Hwan;Karadeniz, Fatih;Lee, Jung Im;Seo, Youngwan;Kong, Chang-Suk
    • Ocean and Polar Research
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    • v.44 no.1
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    • pp.29-38
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    • 2022
  • Halophytes are plants that live in harsh environments in coastal regions and are known for their diverse chemical compositions. Limonium tetragonum, a halophyte endemic to Korean shores, is known for its bioactive compounds and is utilized in folk medicine. In this study L. tetragonum extract (LHE) was used to determine and evaluate its anti-osteoporotic properties. Pre-adipocyte and pre-osteoblasts were induced to differentiate along with LHE treatment, and their differentiation was evaluated using differentiation markers. LHE treatment decreased lipid accumulation in 3T3-L1 preadipocytes during adipogenesis. Results indicated that the LHE treatment also decreased the levels of key adipogenic transcription factors: PPARγ, SREBP1c, and C/EBPα. Enhancing osteoblastogenesis by LHE treatment was confirmed in osteoblastogenesis-induced MC3T3-E1 pre-osteoblasts. Cells treated with LHE resulted in increased calcification and alkaline phosphatase (ALP) activity compared with osteoblasts without LHE treatment. Pro-osteogenic and anti-adipogenic effects were also confirmed in D1 murine mesenchymal stromal cells which are capable of differentiation into both adipocytes and osteoblasts. LHE hindered adipogenesis and enhanced osteoblastogenesis in D1 MSCs in a similar fashion. In conclusion, L. tetragonum is believed to possess the potential to be utilized as a nutraceutical ingredient against osteoporotic conditions.

Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments

  • Lewis-Lujan, Lidianys Maria;Rosas-Burgos, Ema Carina;Ezquerra-Brauer, Josafat Marina;Burboa-Zazueta, Maria Guadalupe;Assanga, Simon Bernard Iloki;del Castillo-Castro, Teresa;Penton, Giselle;Plascencia-Jatomea, Maribel
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.989-1002
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    • 2022
  • Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.