Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery     

Nora Denk  Vikram Misra  Lynne S. Sandmeyer  Bianca B. Bauer  Jaswant Singh  George W. Forsyth  Bruce H. Grahn 《Canadian journal of veterinary research》2015,79(1):31-38

The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.  相似文献   

Comparative Expression Analysis of Gametogenesis‐Associated Genes in Foetal and Adult Bubaline (Bubalus bubalis) Ovaries and Testes          下载免费PDF全文

SM Shah  N Saini  S Ashraf  M Zandi  MK Singh  RS Manik  SK Singla  P Palta  MS Chauhan 《Reproduction in domestic animals》2015,50(3):365-377

This study was conducted to identify and analyse the expression of gametogenesis‐associated genes and proteins in foetal and adult buffalo gonads of both the sexes. Relative quantification of the genes was determined by qPCR and Western blotting. Immunohistochemistry was also performed for various gametogenesis‐associated proteins in foetal and adult gonads of both the sexes. We observed significantly (p < 0.05) increased expression of primordial germ cell‐specific, meiotic as well as genes associated with oocyte maturation and development in foetal ovaries as compared to the adult ones. However, significantly (p < 0.05) increased expression of proteins associated with oocyte maturation like GDF9 and ZP4 was found in adult ovaries, indicating temporal regulation of mRNA translation during oogenesis. Meiotic genes showed significantly (p < 0.05) increased expression in adult testes as compared to foetal testes and ovaries, indicating onset of meiosis at a later stage in spermatogenesis. In general, the expression of primordial germ cell‐associated as well as meiotic genes was higher in adult testes, indicating the increased biological activity in the organ. Immunohistochemistry revealed localized expression of gametogenesis‐associated proteins in ovarian follicles and seminiferous tubules of testes, while the surrounding somatic tissues were devoid of these proteins. The study gives an understanding of the sequential and temporal events of gene expression as well as mRNA translation during male and female gametogenesis. It could also be concluded that follicles and seminiferous tubules are the functional units of the female and male gonads, respectively, and their function could be enhanced by appropriate chemical and genetic intervention of the somatic tissue immediately surrounding them. This assumes importance in the context that buffalo attains sexual maturity at an older age of 2–3 years and have smaller ovaries with lesser number of primordial follicles in comparison with cattle, which is suggested to be the main reason of their poor breeding performance.  相似文献   

Baicalein induction of hydroxyl radical formation via 12-lipoxygenase in human platelets: an ESR study     

Chou DS  Lee JJ  Hsiao G  Hsieh CY  Tsai YJ  Chen TF  Sheu JR 《Journal of agricultural and food chemistry》2007,55(3):649-655

The pro-oxidant activities of baicalein, morin, myricetin, quercetin, and rutin were examined in various cell-containing systems including human platelets, rat vascular smooth muscle cells, human umbilical vein endothelial cells (HUVECs), human THP-1 cells, and fibroblast cells. Electron spin resonance (ESR) results showed that only baicalein generated hydroxyl radicals in a resting human platelet suspension, whereas the other flavonoids showed no effects on any of the resting cell systems. A low concentration of arachidonic acid (AA) increased the intensity of hydroxyl radicals, but a high concentration inhibited it. Collagen and thrombin, platelet aggregatory agents that can cause the release of AA by platelets, enhanced baicalein-induced hydroxyl radical formation, whereas ADP and U44619 showed no significant effects. Quinacrine and 5,8,11,14-eicosatetraenoic trifluoromethyl ketone, both PLA2 inhibitors, significantly attenuated baicalein-induced hydroxyl radical formation. These results suggest that baicalein-induced hydroxyl radical formation is associated with AA metabolite enzymes in human platelets. The formation of hydroxyl radicals was significantly inhibited by lipoxygenase inhibitors including nordihydroguaiaretic acid, (-)-epicatechin, (-)-epicatechin gallate, and hinokitiol, but was not affected by desferroxamine or the heme protein inhibitors KCN and NaN3. On the other hand, semiquinone free radicals were generated when baicalein was incubated with horseradish peroxidase/H2O2 or platelets/AA. The semiquinone radicals formed in the platelets/AA system could be extensively inhibited by desferroxamine, diethylenetriaminepentaacetic acid, KCN, and NaN3, indicating that prostaglandin H synthase (PGHS)-peroxidase may be involved. The results of this study led to the proposal that baicalein induces hydroxyl radical formation via 12-lipoxygenase and induces semiquinone radical formation via PGHS-peroxidase in human platelets.  相似文献   

Improved HPLC-MS/MS method for determination of isoxaflutole (balance) and its metabolites in soils and forage plants     

Lin CH  Lerch RN  Garrett HE  Li YX  George MF 《Journal of agricultural and food chemistry》2007,55(10):3805-3815

A robust multi-residue procedure is needed for the analysis of the pro-herbicide isoxaflutole and its degradates in soil and plant materials at environmentally relevant (<1 microg kg-1) levels. An analytical method using turbo-spray and heat-nebulizer high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the analysis of isoxaflutole (IXF) and its two metabolites, diketonitrile (DKN) and the benzoic acid metabolite (BA), at sub-microgram per kilogram levels in soil and plant samples. The average recoveries of the three compounds in spiked soil and plant samples ranged from 84 to 110% and 94 to 105%, respectively. The limits of quantification were validated at 0.06 microg kg-1 for soil and 0.3 microg kg-1 for plant samples. The limits of detection (LOD) for soil analysis were 0.01, 0.002, and 0.01 microg kg-1 for IXF, DKN, and BA, respectively. Corresponding LOD for the plant analysis method were 0.05, 0.01, and 0.05 microg kg-1. The developed method was validated using forage grass and soil samples collected from a field lysimeter study in which IXF was applied to each of four forage treatments. Forage plants and soils were sampled for analyses 25 days after IXF application to the soil. In soils, IXF was not detected in any treatment, and DKN was the predominant metabolite found. In forage plants, the concentrations of DKN and BA were 10-100-fold higher than that in soil samples, but IXF was not detected in any forage plants. The much higher proportion of BA to DKN in plant tissues (23-53%), as compared to soils (0-5%), suggested that these forages were capable of detoxifying DKN. The developed methods provided LODs at sub-microgram per kilogram levels to determine the fate of IXF and its metabolites in soils and forage plants, and they also represent considerable improvements in extraction recovery rates and detection sensitivity as compared to previous analytical methods for these compounds.  相似文献   

Effect of dips in a 1-methylcyclopropene-generating solution on 'Harrow Sun' plums stored under different temperature regimes     

Manganaris GA  Vicente AR  Crisosto CH  Labavitch JM 《Journal of agricultural and food chemistry》2007,55(17):7015-7020

The effect of postharvest dips in a 1-methylcyclopropene-generating solution of the formulation AFxRD-038 (Rohm & Haas) on plum fruit (Prunus salicina Lindell cv. 'Harrow Sun') quality and ripening during storage was determined. Fruit weight loss, tissue firmness, soluble solids content (SSC), titratable acidity (TA), ethylene production, respiration, and the activities of the cell wall modifying enzymes polygalacturonase (PG), 1,4-beta-D-glucanase/glucosidase (EGase), beta-galactosidase (beta-gal), and pectin methylesterase (PME) were measured. Fruit reddening, anthocyanin content, and phenylalanine ammonia-lyase (PAL) activity were also analyzed. The 1-MCP-treated fruit showed reduced ethylene production and respiration rate and delayed softening, which was associated with the reduction in the activity of PG, EGase, and beta-gal. The immersion in 1-MCP-generating solutions also decreased weight and acidity loss without modifying the fruit SSC. The immersion treatment was particularly effective in the fruit stored at 5 degrees C, keeping higher overall quality, maintaining lower levels of anthocyanins and PAL activity, and preventing flesh reddening. The present data show that beneficial effects in delaying plum fruit ripening and controlling chilling injury can be obtained by dipping the fruits in a solution of this novel 1-MCP-generating formulation.  相似文献   

Determination of anabolic steroids in muscle tissue by liquid chromatography-tandem mass spectrometry   总被引：1，自引：0，他引：1  

Kaklamanos G  Theodoridis G  Papadoyannis IN  Dabalis T 《Journal of agricultural and food chemistry》2007,55(21):8325-8330

A specific and sensitive method based on liquid chromatography-tandem mass spectrometry using atmospheric pressure chemical ionization (LC-APCI-MS/MS) has been developed for the determination of four anabolic steroids [trenbolone, methylboldenone, methyltestosterone, and norethandrolone] in bovine muscle. Methyltestosterone- d 3 was used as internal standard. The procedure involved enzymatic hydrolysis, extraction with tert-butyl methyl ether, defattening, and final cleanup with solid-phase extraction with Oasis HLB cartridges. The analytes were analyzed by reversed-phase LC-MS/MS, acquiring two diagnostic product ions from the chosen precursor [M + H] (+) for the unambiguous confirmation of hormones. The method was validated according to the European Commission Decision 2002/657/EC for the detection and confirmation of residues in products of animal origin. The limits of detection (LOD) and limits of quantitation (LOQ) were found to be 0.3 ng/g and 1.0 ng/g, respectively. The accuracy and precision have been determined, with recoveries ranging from 83% to 104% and the CV factor not exceeding the value of 7%. The decision limits CCalpha were calculated and ranged from 0.05 to 0.15 ng/g while the detection capabilities CCbeta ranged from 0.09 to 0.25 ng/g. The method proved to be sensitive and reliable and thus renders an appropriate means for residue analysis studies.  相似文献   

Autoimmune hepatitis-specific antibodies against soluble liver antigen and liver cytosol type 1 in patients with chronic viral hepatitis     

Eirini I Rigopoulou  Maria Mytilinaiou  Ourania Romanidou  Christos Liaskos  George N Dalekos 《Journal of Autoimmune Diseases》2007,4(1):2

  相似文献   

Effect of protein source and level on growth and performance of the captive freshwater prawn,Macrobrachium rosenbergii     

George H. Balazs  Ernest Ross 《Aquaculture (Amsterdam, Netherlands)》1976,7(4):299-313

Growth, feed conversion and survival were determined for juvenile Macrobrachium rosenbergii held in tanks under semi-controlled environmental conditions. Feeding trials incorporated water-stable diets at three levels of protein (15, 25 and 35%). The principal protein source combinations consisted either of soybean and tuna meal, or of soybean, tuna and shrimp meal. In a 244-day comparison of these diets, higher protein content produced larger prawns (P < 0.01), but differences between sources were not significant. No significant differences existed between feed conversion ratios (range 1.36–1.72) or percentage survival (range 90.3–93.6%). Trials of several other diets were also conducted, including soybean and Tilapia meal, and copra and Tilapia meal (25% protein level) as principal protein source combinations. After 167 days on these diets, growth was inferior to that obtained with soybean and tuna meal or soybean, tuna and shrimp meal combinations. No significant differences existed between feed conversion ratios or percentages of survival.For the 244 days, a control group of prawns received no formulated diet. Growth and survival in this group during the first 110 days suggested that naturally occurring algae contributed substantially to the prawns' nutrition.Mean prawn length after 244 days on the best diet (35% protein from soybean and tuna meal) was 73 mm, and growth rate was equivalent to that achieved under pilot pond conditions.  相似文献   

The Chinese grass carp, Ctenopharyngodon idella, its biology,introduction, control of aquatic macrophytes and breeding in the Sudan     

T.T. George 《Aquaculture (Amsterdam, Netherlands)》1982,27(3):317-327

The paper describes the distribution and biology of the grass carp following its introduction into Sudan. It provides aspects on the culture of the grass carp and its breeding under Sudanese conditions with emphasis on future research.  相似文献   

VH1-44 gene usage defines a subset of canine B-cell lymphomas associated with better patient survival     

Hsiao-Wei Chen  George W. Small  Alison Motsinger-Reif  Steven E. Suter  Kristy L. Richards 《Veterinary immunology and immunopathology》2014,157(3-4):125-130

The use of specific immunoglobulin heavy chain variable region (VH) genes has been associated with increased patient survival in human B-cell lymphomas (hBCL). Given the similarity of human and canine BCL (cBCL) in morphology and clinical treatment, we examined the choice of VH in cBCL and determined whether VH gene selection was a distinct feature associated with survival time in dogs. VH gene selection and mutational status in 52 cBCL, including 29 diffuse large B-cell lymphomas (cDLBCL, the most common subtype of cBCL), were analyzed by comparison with the 80 published canine germline VH gene sequences. We further examined the prognostic impact of the subgroups defined by these features on canine survival. We found that VH1-44 was preferentially expressed in the majority of the 52 cBCLs (60%) as well as in the majority of the cDLBCL subset (59%). VH1-44 gene expression was associated with a statistically better overall survival (p = 0.039) in cBCL patients, as well as in the cDLBCL subset of patients (p = 0.038). These findings suggest that VH gene selection in cBCL is not random and may therefore have functional implications for cBCL lymphomagenesis, in addition to being a useful prognostic biomarker.  相似文献