93 research outputs found
Use of Primary Quail Embryo Fibroblast cells for propagation and assay of Avian viruses
No full textA primary fi broblast cells from embryos of brown quail Coturnix ypsilophora has been established and partially characterized. The cells were maintained in Modifi ed Eagle’s medium (MEM) supplemented with 10% fetal bovine serum. The cells were able to grow at temperatures between 35°C and 38°C with optimum temperature of 37°C. The growth rate of primary quail fi broblast cells increased as the FBS proportion increased from 5% to 20% at 37°C with optimum growth at the concentrations of 10% or 15% FBS. The cells showed no microbial contamination throughout the period of experiment and the total chromosome number of a diploid cell was 78, according to karyotyping and chromosome analysis. The susceptibility of quail primary cells for avian viruses was investigated in this study after inoculation with ND and IB viruses. Both viruses showed a satisfactory CPE development and the infectivity were assayed by virus titration (TCID₅₀). This suggests that the quail primary cells can be used for isolation of various avian viruses with further steps of infectivity confi rmation in the future.Mayada F. Hasoon, Majed H. Mohammed and Jaladet M.S. Jubrael (Mauida, Al Khallawi
A study of the bone marrow derived mesenchymal stromal cells in rats - proliferation and immunophynotypic markers
No full textMesenchymal stem cells
derived from bone marrow (BM-MSCs) have
the ability to differentiate into multiple cell
lineages. Although the cultivation of these
cells has led to a number of characterisation
studies, some significant morphological
and immunohistochemical properties
are still lacking. In this study, isolation of
BM-MSCs, morphological features, cell
viability, immunophenotypic properties
and cryopreservation of BM-MSCs were
examined in detail. The results demonstrate
that the cells isolated from BM-MSCs were
plastic adherent and had fibroblastic spindle
shape after three passages and get confluent
monolayer cells 70-80% after 4-7 days
post-subculture. Based on the cell viability
analysis, the BM-MSCs showed an increase
in cell viability starting from passage 1 until
passage 10. Immunophenotypic analysis
demonstrated that BM-MSCs were positive
for CD44 and CD105 and negative for CD34.
Functional analysis of cryopreservation of
BM-MSCs from P6 after 6 months expressed
good proliferation rate and cell viability
Isolation and characterization of viral nervous necrosis virus on newly established marine fish cell line and its pathogenicity in non-marine host
Get PDFIn Southeast Asia, the intensification of aquaculture industry has led to the occurrence of various infectious diseases. Virus diseases in particular, affecting especially the larval and juvenile stages, are the most suppressing factors for sustainable production of high value fish species such as seabass (Lates ca/carifer). Inevitable, because of lack of sensitive methods that could be used for diagnosis and the inadequacy of pathogen control, the aquaculture industry will continue to be plagued by these diseases. Currently, viral nervous necrosis (VNN) is one of fish diseases listed by the Office International des Epizooties (OlE) as a notifiable disease in the production of marine fish worldwide. Thus the main objective of the present study was to determine the role of VNN virus (VNNv) as a pathogen in the Malaysian aquaculture industry, with emphasis on virus isolation in susceptible cell line, virus identification using polymerase chain reaction (PCR) and identification of characteristic microscopy lesions in VNNv infections. A new cell line designated as Asian Seabass Brain (ASBB-1) was derived from the brain tissue of seabass cultured in Malaysia. This cell line was maintained in Leibovitz L-15 media supplemented with 10% fetal bovine serum (FBS). The ASBB-1 cell line was sub-cultured more than 60 times over a period of 15 months. This cell line consists predominantly of fibroblastic-like cells, which are able to grow at temperatures between 20°C and 30°C with an optimum temperature of 25°C. At 25°C, the growth rate of these cells increased as the proportion of FBS increased from 5 to 20% with optimum growth at 15% or 10% FBS. The ASBB-1 cell line was characterized by karyotyping, and chromosome number distributions were different with subcultures P20 and P50 producing 48 and 46 chromosome peaks respectively. The ASBB-1 cells showed 90% viability after recovery from one year storage in liquid nitrogen. The ASBB-1 cells up on characterization by peR using primer sets of microsatellite markers of Asian seabass (Lates ca/carifer) were shown to be similar to the cells from tissues of seabass at 250 bp. The VNNv isolate derived from seabass was tested positive for VNN by the 102000 Kit®. The isolate designated UPM08-1 M was used in the cell line infectivity study. The susceptibilities of ASBB-1 cell line isolate and the highly permissive commercial SSN-1 cells to UPM08-1 M were compared. The results showed that ASBB-1 cell line was susceptible to VNNv (RGNNV genotype) with typical cytopathic effect (CPE) manifesting mainly as rounding-up at1 day post infection (dpi), severevacuolationwithin3-5dpi and complete detachment within 7dpi. The VNNv-induced CPE was further elucidated by electron microscopy (EM). Under EM, the ASBB-1cellsexhibitedvacuolated degeneration with presence of viral inclusion-like bodies. The CPE of VNNv on ASBB-1cells was producedat a virus titer of 109 . 5 TCID50/ml. This indicated that the ASBB-1 cell line is highly susceptible for use in the isolation of VNNv. Biophysical and biochemical characterization of VNNv isolate was determined by heat treatment, UV irradiationand the stability under effect of chemical disinfectants. The VNNv isolate showed susceptibility to heat treatment at 60°C within 30 minutes with no viable virus after 1 hour. UV irradiation at an intensity of 440 mWrcm2 resulted in a reduction in virus titer after 8 minutes. The virus appeared relatively resistant to changes in pH ranging between 2 to 11 after 1 hour incubation at 25°C. Treatment with 2% formalin was not totally effective even after 6 hours. Iodine did not inactivate the virus. Molecular characterization of the tissue culture-propagated virus using RT-PCR and nested peR, showed positive amplifications of genome size of 460 bp and 220 bp respectively. The 220 bp nucleotides from the T4 region of the coat protein gene was sequenced and the phylogenetic analysis resultsshowed close resemblance between the UPM08-1 M strain andSBNNV and GPNNV2strains of Malaysianisolates.The local isolate of UPM08-1 M also showed phylogenetic similarity with other strains from South East Asia, while showed relatively distant similarity with other VNNv strains from the Middle East. Cold water VNNv strains isolated from Norway, Canada and USA were distant phylogenetic relations to the UPM08-1 M strain. Experimental infectivity study was also performed using fresh water guppy fish (Poecilia reticulata) as a model. Guppies were intra nasally infected with 0.1 ml of the UPM08-1M strain at a titre of 109.5 TCID5010.1ml. Although the artificial infection resulted in slight mortality (11%) occurring within 14 days post inoculation, the VNN-specific lesions such as necrosis and vacuolation in the target organs of brain and retina were evident. The presence of virus in infected brain and retina tissues was confirmed by transmission electron microscopy (TEM). The VNNv from experimentally infected guppy was successfully reisolated in ASBB-1 cells inoculated with homogenate of brain and retinas of the survivors. There was gradual increment of virus titer in the guppy throughout the experimental period. The results indicated that guppy fish could be infected with VNNv isolated from diseased seabass. In summary, the study successfully isolated VNNv using the ASBB-1cell line. Complete CPE of this cell line was observed within 5 - 7 dpi after infection with VNNv at producing considerable higher titer of 1095TCID5010.1ml. Thus ASBB-1 cells can be recommended for VNNv (RGNNV genotype) isolation and will be an important tool for future conduct of fish health assurance programs in the aquaculture industry. The study also showed that RT-PCR followed by nested- PCR is a sensitive technique for identification of VNNv isolated from sea bass. It was also shown that guppy fish could be experimentally infected with VNNv, thus can be used as a virus-infection model
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Surfactant-Assisted Growth of CdS Thin Films for Photovoltaic Applications
Get PDFA common non-ionic surfactant, Triton X-100, was used to modify the chemical bath deposition (CBD) of CdS "buffer" layers on Cu(In,Ga)Se2 (CIGS) thin films. X-ray photoelectron spectroscopy (XPS) and Auger electron spectroscopy (AES) data demonstrate that films produced with the surfactant have about the same levels of impurities as films grown without it. For thin, ~130 ..ANG.. CdS layers and relative to devices made without the surfactant, average absolute cell efficiencies were increased from 10.5% to 14.8%, or by a relative 41%. Visual inspection of the CdS depositions reveals one possible mechanism of the surfactant's effects: bubbles that form and adhere to the CIGS surface during the CBD reaction are almost completely eliminated with the addition of the TX-100. Thus, pinholes and thin areas in the CdS layers caused by poor wetting of the substrate surface are sharply reduced, leading to large increases in the open circuit voltage in devices produced with the surfactant
Performance evaluation and optimization of dual-stage L-band EDFA utilizing short gain medium
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Rimegepant for the treatment of migraine
Get PDFMigraine is a common form of primary headache, affecting up to 1 in every 6 Americans. The pathophysiology is an intricate interplay of genetic factors and environmental influence and is still being elucidated in ongoing studies. The trigeminovascular system is now known to have a significant role in the initiation of migraines, including the release of pain mediators such as CGRP and substance P. Traditional treatment of migraine is usually divided into acute and preventive treatment. Acute therapy includes non-specific therapy, such as NSAIDs and other analgesics, which may provide relief in mild to moderate migraines. 5-HT1 agonists may provide relief in severe migraine, but are not universally effective and carry a significant side-effect profile with frequent redosing requirement. Prophylactic therapy may reduce the occurrence of acute migraine attacks in selected patients, but does not completely eliminate it. More recently, CGRP antagonism has been studied and shown to be effective in both abortion and prevention of migraine. Novel medications, targeting CGRP, divide into CGRP antibodies and receptor antagonists (gepants). Rimegepant, a second-generation gepant, has shown efficacy in several clinical trials in treating acute migraine. Ongoing trials are also evaluating its role in migraine prophylaxis, and results are promising. It is also generally safer for use than existing options, does not appear to increase the chance of developing chronic migraines, and carries a very tolerable side effects profile. It is a part of a growing arsenal in migraine treatment, and may present the silver bullet for treatment of this disease
The Characterization Of Chemical Bath Deposited Cds On Single Crystal InP Substrates
No full textAbstractWe have investigated CdS thin films as possible passivating window layers for InP. The films were deposited on single crystal InP by chemical bath deposition (CBD). The film thickness, as optically determined by ellipsometry, was varied from 500 to 840Å. The film morphology was investigated by high resolution scanning electron microscopy (SEM), whereas the film microstructure was studied by X-ray diffraction (XRD) and cross-sectional transmission electron microscopy (TEM). Most of the films were fine-grained polycrystalline CdS, with some deposition conditions resulting in epitaxial growth. Cross-sectional TEM examination revealed the presence of interface contaminants. The effect of such contaminants on the film morphology and microstructure was studied, and various approaches for InP surface cleaning/treatment were investigated. The epitaxial films were determined to be hexagonal on both the (111) and (100) InP substrates; however, they were heavily faulted.</jats:p
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