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Publication Open Access Quantitative evaluation of cellular intensity in cytologic staining over difference time period of post air-dried smear in canine mammary gland tumor(2018) Jeerasak sri-in; Pattita Ruayaree; Panpaga Sangsuriya; Panop wilainam; Parin Suwannaprapha; จีรศักดิ์ ศรีอินทร์; ปทิตตา รวยอารี; พรรณพงา เเสงสุริยะ; ภานพ วิไลนาม; ปริญ สุวรรณประภา; จุฬาลงกรณ์มหาวิทยาลัย. คณะเภสัชศาสตร์; มหาวิทยาลัยมหิดล. คณะสัตวแพทยศาสตร์. ภาควิชาปรีคลินิกและสัตวศาสตร์ประยุกต์Diagnostic cytology is an initial laboratory testing to diagnose various types of cancer, infections and inflammatory diseases. The collection and processing of cytological specimens are very important to produce good quality specimensPublication Open Access Quality and quantity of Asian elephant (Elephas maximus) peripheral blood mononuclear cells isolated by polymer of sucrose or colloidal silica solution gradient centrifugation(2008) Porntep Gittipongsiri; Somphon Makpun; Teeraphong Phansod; Dulyatad Gronsang; Roschong Boonyarittichaikij; Witthawat Wiriyarat; Mahidol University. Faculty of Veterinary SciencePeripheral blood mononuclear cells (PBMCs) are useful in cellular immunology, infectious disease diagnosis and molecular genetics research. This study was aimed to determine an appropriated procedure for PBMCs isolation from an Asian elephant (Elephas maximus) by using gradient centrifugations method. The efficacy of four different density solutions including polymer of sucrose solution (Ficoll-Hypaque mixture) of density 1.077 and colloidal silica solution (Percoll) of density 1.077, 1.078 and 1.079 g/cm3 were compared in elephant PBMC separation. Quality and quantity of isolated PBMCs were estimated by using five parameters including cell purity, cell recovery rate, cell viability, viability of PBMCs after 5 days cultivation and total cell count. The results showed that the total cell count was significantly different among 4 treatments, in which Percoll with density 1.079 g/cm3 yielded the highest PBMCs (average = 3.94 X 107 cells/cm3) and highest PBMCs recovery rate (average = 82.61%). However, the purity, viability and viability of PBMCs after 5 days cultivation were not significantly different (P<0.05). In conclusion, Percoll with a density of 1.079 g/cm3 is the most suitable density gradient treatment for Asian elephant PBMCs isolation in both quantity and quality aspects.Publication Open Access Effects of the three flavonoids; kaempferol, quercetin, and myricetin on Baby hamster kidney (BHK-21) cells and Human hepatocellular carcinoma cell (HepG2) cells proliferations and total Erk1/2 protein expression(2017) Sookruetai Boonmasawai; Arpron Leesombun; Kridsada Chaichoun; Jarupha Taowan; Ladawan Sariya; Orathai Thongjuy; Mahidol University. Faculty of Veterinary ScienceKaempferol, quercetin, and myricetin were regarded as the potential therapeutic flavonoids in several types of cancers. And the cancer cell proliferations that were directly induced by MAPK/Erk signaling pathway had been the important target of cancer treatment. Thus, this study was investigated the anti-proliferative effects of these three flavonoids together with total Erk1/2 protein expressions on Human hepatocellular carcinoma cell (HepG2) and Baby hamster kidney cells (BHK-2). The data showed the cytotoxic effects of 100 μM kaempferol, 5 μM myricetin and 50 μM quercetin on BHK-21 cells at 24h. After same incubation time, kaempferol (5 μM), myricetin (1 μM) and quercetin (1 μM) could significantly inhibit HepG2 cell proliferations. By western blot analysis, kaempferol and myricetin could not affect Total Erk1/2 proteins expression. But quercetin obviously suppressed total Erk1/2 protein expression in HepG2 cells at 24h. In conclusion, all three flavonoids had significant inhibitory effects on Hepatocellular carcinoma cells without cytotoxic effects on normal fibroblast. And inhibitory effects of quercetin in cancer cell proliferation related to total Erk1/2 protein reduction.Publication Open Access การพัฒนา Diploid cells จากเซลล์เหงือกของปลาคาร์พ เพื่อใช้ในการแยกเชื้อไวรัสคอยเฮอร์ปีส์(2555) น้ำอ้อย เถาวัลย์; นัทธภัทร เกตุฉิม; สมจิตร ใช้วัฒนรุ่งเรืองไพศาล; ปรุศก์ สุกใส; แพรวพร ไทยจงรักษ์; กฤษฎา ใจชื้น; มหาวิทยาลัยมหิดล. คณะสัตวแพทยศาสตร์. ศูนย์เฝ้าระวังและติดตามโรคจากสัตว์ป่า สัตว์ต่างถิ่นและสัตว์อพยพ; มหาวิทยาลัยมหิดล. คณะสัตวแพทยศาสตร์. ภาควิชาเวชศาสตร์คลินิกและการสาธารณสุข; มหาวิทยาลัยมหิดล. คณะสัตวแพทยศาสตร์. ภาควิชาปรีคลินิกและสัตวศาสตร์ประยุกต์. In this study, KHV was cultured consecutively in koi gill (KG) cells of passage 29 - 30 and 33 - 34. The fluid from each passage was tested for KHV by polymerase chain reaction technique and positive result was found in both passages. Thus, KG cells canPublication Open Access Using of CRFK and HeLa Dual Cell-Culture System to Isolate Canine Distemper Virus(2020) Jarupha Taowan; Pruksa Julapanthong; Wanvisa Surarith; Natthapat Ketchim; Chalisa Mongkolphan; Kridsada Chaichoun; Mahidol University. Faculty of Veterinary Science; Mahidol University. Faculty of Veterinary Science. The Monitoring and Surveillance Center for Zoonotic Disease in Wildlife and Exotic Animals; Mahidol University. Faculty of Veterinary Science. Internal Medicine Unit Prasu Arthorn Animal HospitalCanine distemper virus (CDV) is a common infectious agent that affects respiratory and nervous systems in canine, pet and wildlife worldwide. The virus isolation using cell culture is one of laboratory diagnostic methods for detection of virus infections. Many different types of cultured cells can be used for isolation of distemper virus, which provides the different outcome. This study aimed to evaluate the efficacy of dual cell culture system, Crandell-Rees feline kidney (CRFK) and HeLa cells for virus isolation. A total of 13 clinical specimens were taken by nasal and oropharynx swab from dogs with clinical signs compatible with CDV infection. These specimens were positively confirmed CDV by using Reverse transcriptase - polymerase chain reaction (RT-PCR) for viral genome detection. The CDV RT-PCR positive specimens were then taken to isolate in both cell-lines for five passages, and were confirmed by RT-PCR again. By using CRFK and HeLa cells, the successful viral isolation with CDV positive specimens was recorded at 7/13 (53.84%) in CRFK and 6/13 (46.15%) in HeLa cells, respectively. The dual cell-culture system showed an increase of the successful isolation at 8/13 (61.53%). Our results suggested that the application of dial cell-culture system was more effective than using single cell-culture for isolation of canine distemper virus.Publication Open Access Loop-mediated Isothermal Amplification (LAMP): An Alternative Molecular Diagnosis(2011) Dusit Laohasinnarong; Mahidol University. Faculty of Veterinary Science. Clinical Sciences and Public Health DepartmentPublication Open Access Urinary markers in Babesia canis vogeli-infected dogs(2016) Sivapong Sungpradit; Rapeeporn Pikhroh; Wanvisa Thanasaksakul; Mahidol University. Faculty of Veterinary Science. Department of Pre-clinic and Applied Animal ScienceCanine babesiosis, a life-threatening tick-borne blood parasitic disease in dogs, caused by Babesia canis vogeli, is a health problem in companion animals. The disease causes febrile illness, hemolytic anemia, pre-hepatic jaundice, and thrombocytopenia. Moreover, renal dysfunction from babesiosis has been reported. The purpose of this study was to investigate urinary markers that might be sensitive and specific for the early detection of renal dysfunction in B. c. vogeli-infected dogs. Blood and urine samples were collected from 11 dogs. The blood and urine samples were divided into two groups. B. c. vogeli-infected dogs group including six infected dogs as confirmed by microscopic examination and multiplex polymerase chain reaction. Non-infected dogs group included five healthy dogs. Blood samples were subjected to hematology and biochemistry analysis while urine samples were stored at -80oC until analyzed. Three candidate urinary markers (urinary immunoglobulin G, uIgG; urinary C-reactive protein, uCRP; and urinary retinol-binding protein, uRBP) were examined using commercial enzyme-linked immunosorbent assays (ELISA); two additional candidate markers, aspartate aminotransferase to alanine aminotransferase ratio (AST/ALT) and urinary creatinine to serum creatinine ratio (UCr/SCr) were also studied. The results demonstrated that hemoglobin, red blood cell count, and hematocrit were significantly different between B. c. vogeli-infected dogs and non-infected dogs while the candidate markers were not. In conclusion, the selected candidate markers could not be used as urinary markers for renal dysfunction in B. c. vogeli-infected dogs. However, further study should investigate other urinary markers such as albumin, tubular enzymes, and tubular proteins as well as high-throughput technologies such as the proteomic approach.Publication Open Access Development of specific activity of amylolytic and proteolytic enzymes in duodenum and jejunum of the small intestinal tract of meat-type ducks during 1-42 days of age(2017) Kriengkrai Prahkarnkaeo; Kiattawee Choowongkomon; Boonorm Chomtee; Choawit Rakangthong; Chaiyapoom Bunchasak; Mahidol University. Faculty of Veterinary Science. Department of Clinical Science and Public HealthThis experiment was conducted to elucidate the pattern of amylolytic and proteolytic digestive enzyme activities in meat-type ducks during 1-42 days of age. Twenty-eight male meat type ducks (Cherry Valley strain) were used, and diets based on corn-soybean meal were offered ad lib throughout the experimental period. At duodenal and jejunal segments, the specific activity (SA) of amylolytic (amylase) and proteolytic (trypsin, chymotrypsin and total proteases) enzymes were determined at 1, 7, 14, 21, 28, 35 and 42 days of age. Maximal or nearly maximal SA of all enzymes at first day after hatch (1 day of age) and the activity declined on 7 day of age was found (P<0.05). After 7 days of age, all enzymes tended to reach a nadir at day 14 or day 21, then the SA were significantly increased from 21 to 42 days of age (P<0.05). Therefore, the pattern of SA of all enzymes at duodenal and jejunal segments were changed in a cubic trend (P< 0.01) with age. In ducks, it is indicated that the embryonic enzyme reserve may be a reason of high SA of amylolytic and proteolytic enzymes at 1 day of age, then the SA are activated again after 21 days of age by intestinal development processes and/or increasing amount of feed intake.Publication Open Access Effects of scabraside D on cell viability inhibition and apoptosis promotion of human hepatocellular carcinoma(2018) Rassameepen Phonarknguen; Thanakorn Rawangchue; Orathai Thongjuy; Kanjana Assawasuparerk; รัศมีเพ็ญ โพธิ์นาคเงิน; ธนกร ระวังชื่อ; อรทัย ทองจุ้ย; กาญจนา อัศวศุภฤกษ์; Mahidol University. Faculty of Veterinary Science. The Monitoring and Surveillance Center for Zoonotic Disease in Wildlife and Exotic Animals; Mahidol University. Faculty of Veterinary Science. The Center for Veterinary Diagnosis; Mahidol University. Faculty of Veterinary Science. Department of Pre-Clinical and Appiled Animal ScienceScabraside D, derived from the sea cucumber Holothuria scabra is a sulfated triterpene glycoside which possess in various biological activities. We assessed the activity of scabraside D and their effects on cell viability and apoptosis on human hepatocellular carcinoma (HepG2) cell lines by MTT assay and staining with Hoechst 33342. The 25 to 100 μg/mL dose of scabraside D significantly decreased the viability of HepG2 cells, in a dose-dependent manner. The treatment with scabraside D at dose 50 and 100 μg/mL significantly also induces morphological changes of apoptotic cell, including cell shrinkage, nuclear chromatin condensation and nuclear fragmentation. Quantitative real-time PCR shows that scabraside D up-regulated Bax and Caspase-3 while down-regulated Bcl-2 expressions in the HepG2 cells in dose-dependent manner. In conclusion, scabraside D can inhibit cell viability and induce apoptosis in HepG2 cells. This study show that scabraside D may be used as a new therapeutic agent for human hepatocellular carcinomaPublication Open Access The Effect of adding L-Cysteine in Modified Kenny's extender on the quality of chilled stallion semen at 5oC(2013) Panithi Sukho; Hiran Juwarahawong; Slitwan Sangiam; Kampon Kaeoket; Mahidol University. Faculty of Veterinary Science. Dapartment of Clinical Sciences and Public Health DepartmentThe aim of this study was to investigate the effect of supplementation of L-Cysteine in Modified Kenny's extender on chilled stallion semen quality at 5oC. Semen samples were collected from 3 stallions by artificial vagina and centrifugation to remove seminal plasma and diluted with Modified Kenny's extender to 50 x 106 sperm/ml. Semen samples (n=10 ejaculates) were divided into 3 portions, the first portion as the control (KE), the second portion was added with L-Cysteine 200 μmol/l (KEC200) and added L-Cysteine 400 μmol/l in the last portion (KEC400). In all samples, the percentage of progressive motility, viability and acrosome integrity were evaluated at 0, 24, 48 and 72 h after storage at 5oC. There were a significant (P < 0.05) lower percentage of dead spermatozoa in group KE200 than other groups at 24 h. There was a significantly lower percentage of progressive motility in KEC 400 compared toother groups at 48 h (P < 0.05). In addition, there was a tendency (P = 0.1) of lower percentage of dead spermatozoa in group KE200 than other groups at 48 h. At 72 h, comparing all groups, a higher percentage of viability in groups KEC200 and KEC400 was found, but not statistically significant. In conclusion, Addition of L-cysteine at a concentration of 200 μmol/l to the Modified Kenney's extender tend to improve the stallion chilled semen progressive motility, dead/alive, viability and acrosome integrity during chilled storage at 5oC for 48 h.