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1.
Vet Med Sci ; 9(5): 2132-2143, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37466043

RESUMO

CONTEXT: The nutritional value of enzyme-supplemented wheat-contained diets can be influenced by interactions between wheat cultivar and exogenous enzyme, which in turn cause various effects on the performance of poultry. AIMS: The effects of diets including various wheat cultivars with or without enzyme on productive performance and egg quality characteristics were evaluated. METHODS: A total of 270 58-week-old Lohmann LSL-Lite were allocated to 9 experimental dietary groups with 5 replicate cages and 6 birds per each cage. Based on a 4 × 2 factorial arrangement of treatment, nine iso-caloric and iso-nitrogenous experimental diets, including four wheat cultivars (Sardari, Azar 2, Pishgam and Sirvan) with and without enzymes plus a corn-based control diet, were tested in a completely randomized design during an 8-week trial period. KEY RESULTS: From weeks 58 to 63 and 63 to 67 of age, the hens fed with Sardari-included diet showed the lowest feed intake compared to other groups (p < 0.05). Egg production (EP), feed conversion ratio (FCR) and egg mass (EM) were not influenced by the experimental diets. At the age of 62 weeks, the effect of adding enzyme on egg shape index was significant. Moreover, in sampling week 67, increased eggshell weight (%) was detected in enzyme-supplemented diets compared to diets with no enzyme (10.03 vs. 9.37, respectively). Besides, the shell thickness was the highest in the group fed with Azar 2, and the lowest in the group fed with Sirvan at the age of 67 weeks (p < 0.05). Albumen weights (%) were also significant in some cases. The highest albumen weight was observed in Sirvan-included diet without enzyme on week 62. Reduced yolk weight was detected in enzyme-supplemented Pishgam-included diet on 62 weeks of age (p < 0.05). CONCLUSIONS: The inclusion of the diets of laying hens with the four tested Iranian wheat cultivars has no negative impact on most productive performance and egg quality indices. Moreover, supplementing the wheat-included diets with non-starch polysaccharidase enzyme (beta-endopower) improved eggshell weight on week 67.


Assuntos
Galinhas , Triticum , Animais , Feminino , Irã (Geográfico) , Óvulo , Dieta/veterinária
2.
Anim Biotechnol ; 34(4): 1209-1213, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34928775

RESUMO

This study aimed to investigate the association between polymorphisms of ND1 and CYTB genes and in vitro early embryo development of Sanjabi sheep. Blood and ovarian samples were collected from a local slaughterhouse. The cumulus-oocyte complexes with a diameter greater than 3 mm were aspirated from follicles, and in vitro maturation (IVM) and in vitro culture (IVC) rates of them were recorded. A respective 1200 bp and 980 bp fragments of ND1 and CYTB genes were genotyped using a modified single strand conformation polymorphism (SSCP) method. The results of this study revealed that four different patterns, named as A, B, C, and D were observed for both ND1 and CYTB genes. The ND1 gene polymorphisms had significant effects on the IVM and IVC rate (p < 0.05). The pattern C of the ND1 gene significantly increased the IVM rate compared to the patterns A, B and D. For the IVC, the highest and lowest means were related to the C and B patterns, respectively. The CYTB gene polymorphisms also had significant effects on IVC (p < 0.01), but the IVM did not affected (p = 0.07). Here, the pattern D had the highest and the pattern C had the lowest means for both IVM and IVC rates.


Assuntos
Técnicas de Maturação in Vitro de Oócitos , Oócitos , Feminino , Animais , Ovinos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Desenvolvimento Embrionário/genética , Ovário , Polimorfismo Genético/genética
3.
Vet Med Sci ; 8(4): 1603-1610, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35582985

RESUMO

CONTEXT: Full fat soybean (FFSB) can be used in poultry diets as a source of fat and protein, without extra cost for oil extraction; however, raw FFSB contains several anti-nutritional factors. AIMS: Investigating the effects of replacing dietary soybean meal (SBM) with processed (roasted or autoclaved) FFSB on the performance of laying hens and egg quality traits. METHODS: A total of 324 Bovans White strain laying hens were randomly selected and distributed in 54 replicate cages. Based on a completely randomized design with a 2 × 4 factorial arrangement of treatments plus a control group, nine diets in which SBM were replaced with 25%, 50%, 75% and 100℅ roasted or autoclaved FFSB, with six replicates (with six hens each), were evaluated during an 8-week trial period (60-68 weeks). KEY RESULTS: Significantly increased feed intake (FI) was observed in hens fed diets including autoclaved FFSB (p < 0.05). The interactions between SBM replacement level and processing on feed conversion ratio (FCR), egg production (EP) and egg mass (EM) were significant (p < 0.01). Hens fed the diet with 100% roasted FFSB instead of SBM showed poor performance in terms of FCR, EP and EM during the experimental period. The main effect of SBM-replacing level on Haugh unit was significant (p < 0.05). The main effect of processing on shell thickness was significant (p < 0.05), so autoclaved FFSB caused higher eggshell thickness than control and roasted FFSB. CONCLUSIONS: Replacement of dietary SBM with autoclaved FFSB can improve laying hens' performance in terms of FCR, EP, EM and eggshell quality and yolk colour. IMPLICATIONS: The use of autoclaved FFSB to replace commercial SBM and its oil in the diet of laying hens is recommended. Feed factories can formulate the diets of laying hens with autoclaved FFSB without extra cost due to oil extraction soybeans.


Assuntos
Ração Animal , Glycine max , Ração Animal/análise , Animais , Galinhas/metabolismo , Dieta/veterinária , Feminino , Óvulo
4.
Zygote ; : 1-3, 2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34588087

RESUMO

The aim of this study was to investigate mitochondrial ND5 gene polymorphisms and their relationship with in vitro maturation (IVM) and in vitro culture (IVC) of Sanjabi sheep. Blood and ovarian samples of adult ewes were obtained from a local slaughterhouse. For each ovarian sample, cumulus-oocyte complexes larger than 3 mm in diameter were aspirated from follicles, and their IVM and IVC rates were recorded. A 666-bp fragment of the ND5 gene was amplified using the polymerase chain reaction. The samples were genotyped using a modified single-stranded conformation polymorphism (SSCP) method, and an association study was conducted with IVM and IVC rates. Six different SSCP patterns, designated A, B, C, D, E and F with respective frequencies of 8, 47, 4, 4, 32 and 5%, respectively, were observed. According to the results of association analysis, there was no significant association between the ND1 gene polymorphisms and the IVM and IVC rates (P > 0.05).

5.
Trop Anim Health Prod ; 52(5): 2283-2290, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32144657

RESUMO

Genetic structure and genetic diversity levels of indigenous Iranian sheep breeds are not clear, despite the interest this region has in itself as an important center for domestication of livestock. Early population genetic studies have reported high levels of diversity among Iranian sheep breeds until recently, when high admixture levels and genetic homogeneity have been detected. The rapid reduction of diversity observed in Iranian breeds might be due to an increasing trend of intensive crossbreeding practices or even total replacement of native breeds by highly specialized and productive ones. From a conservative perspective, this situation is highly concerning; thus, it might be wise to consider a conservation program in Iran to preserve the original genetic diversity in native sheep breeds. In this study, a total of 1065 animals with the purest morphological features representing 24 Iranian indigenous sheep breeds were sampled, corresponding to ancestral breed diversity. These samples were genotyped for 17 microsatellite loci in order to (1) determine the native ancestral diversity of Iranian breeds, (2) define the degree of genetic relationship among studied breeds, and (3) assess conservation priorities among defined groups. Our results showed no recent loss of diversity, but high genetic diversity levels for indigenous sheep breeds in Iran. Indeed, the analysis of conservation priorities pointed out the importance of 8 breeds for maintaining Iranian sheep breeds' maximum genetic diversity. Thus, under a genetic perspective, these 8 breeds should be the ones included into conservation programs for restocking endangered areas.


Assuntos
Variação Genética , Ovinos/genética , Animais , Conservação dos Recursos Naturais , Genótipo , Hibridização Genética , Irã (Geográfico) , Repetições de Microssatélites
6.
J Anim Physiol Anim Nutr (Berl) ; 102(6): 1521-1532, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30255521

RESUMO

The effects of dietary supplemental lysophospholipids (LPLs) and vitamin C (VC) on performance, activity of antioxidant enzymes, and thyroid hormones of broiler chickens reared under thermoneutral and high ambient temperatures were evaluated. A total of 1,680 broiler chicks (Cobb 500) in finishing rearing period (days 21-38 of age) were allotted to two groups: thermoneutral (TN) and heat stress (HS). In the TN group, 480 chicks were subjected to four treatments with four replicates (n = 30) and maintained in usual ambient temperature (24 ± 1°C). In HS group, the remaining 1,200 chicks were subjected to four treatments with 10 replicates (n = 30) and exposed to high ambient temperature (34 ± 1°C for 8 hr daily). In both groups, four iso-caloric and iso-nitrogenous experimental diets based on a 2 × 2 factorial arrangements including supplemental LPLs (0 or 1,000 mg/kg) and VC (0 or 500 mg/kg) were formulated and used. Supplemental LPLs decreased (p < 0.05) body weight gain and increased FCR in the TN and HS groups. In the TN group, increased (p < 0.05) serum glucose was observed in chickens fed with dietary supplemental VC. In the HS group, decreased (p < 0.05) total protein concentration was detected in birds fed with supplemental LPLs. In both TN and HS groups, decreased (p < 0.05) uric acid concentration was detected in chicks fed with the VC-supplemented diets. A significant (p < 0.05) interaction between LPLs and VC on lactate concentration in the TN group was observed. In the HS group, decreased breast malondialdehyde concentration was detected in birds fed with the VC-supplemented diet. In the TN group, increased serum total antioxidant status was detected in birds fed with the LPLs-supplemented diet. In conclusion, LPLs improved oxidative stability and increased the antioxidant capacity of the serum. In addition, vitamin C modified heat stress and reduced lipid peroxidation.


Assuntos
Antioxidantes/metabolismo , Ácido Ascórbico/farmacologia , Galinhas/sangue , Temperatura Alta , Peroxidação de Lipídeos/efeitos dos fármacos , Lisofosfolipídeos/farmacologia , Criação de Animais Domésticos , Animais , Ácido Ascórbico/administração & dosagem , Galinhas/fisiologia , Suplementos Nutricionais , Feminino , Lisofosfolipídeos/administração & dosagem , Masculino , Malondialdeído/sangue , Malondialdeído/metabolismo , Músculo Esquelético/metabolismo , Distribuição Aleatória , Hormônios Tireóideos/sangue
8.
Anim Reprod Sci ; 179: 49-56, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28214066

RESUMO

The purpose of this study was to investigate leptin gene polymorphisms and their relationships with the characteristics of sperm quality and testicular dimensions. Semen samples were collected from 96 Sanjabi rams during autumn and spring seasons over two years. Simultaneously, the dimensions of length, width and scrotal circumference were measured. Blood samples were taken from the jugular vein to extract DNA. PCR was performed to amplify a 463bp fragment including exon 3 of leptin gene. PCR products were digested by Bcnl and Cail restriction enzymes to identify 170G>A and 332G>A mutations in exon 3, respectively. Leptin gene polymorphism in 170G>A locus had an effect on individual motility trait, water test and scrotal circumference (P<0.05) and animals with the AA genotype had the highest individual motility compared with the GG and GA genotypes (P<0.05). The AG genotypes had the highest water test compared with the GG and AA genotypes (P<0.05) but GG genotype had higher scrotal circumference than that of GA and AA genotypes (P<0.05). The results showed that polymorphism in 332G>A locus had a significant effect on viability trait, water test and scrotal circumference as GA genotypes had the highest amounts for these traits compared with GG genotypes (P<0.05). Based on our knowledge, the current study is the first report on the association of leptin gene polymorphisms with sperm fertility and testicular dimensions in sheep, which suggests leptin gene as a potential gene to be used in breeding programs in order to improve fertility in herds.


Assuntos
Leptina/metabolismo , Polimorfismo Genético/genética , Análise do Sêmen/veterinária , Ovinos/fisiologia , Testículo/anatomia & histologia , Animais , Genótipo , Leptina/genética , Masculino , Reação em Cadeia da Polimerase , Polimorfismo Genético/fisiologia , Estações do Ano , Ovinos/anatomia & histologia , Espermatogênese
9.
South Med J ; 107(7): 437-42, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25010586

RESUMO

BACKGROUND: Primary care physicians (PCPs) are an important part of the decision-making process in the care of patients with cancer. The survey discussed herein evaluates what percentage of academic and community PCPs recognize benefits from systemic therapy in lung and breast cancers. METHODS: PCPs were surveyed regarding their beliefs toward systemic therapy in early- and late-stage lung and breast cancers and were asked to rate the importance of specific factors that influence their referral decisions. RESULTS: A total of 3444 surveys were distributed, and 316 physicians (9.1%) responded: 89 academic physicians (28%) and 227 nonacademic physicians (72%). The rate of returned surveys was equal by specialty. A total of 57%, 42.1% in lung cancer and 72.6 % in breast cancer (P < 0.001) of PCPs, believe in the curative effect of systemic therapy in early stages. Sixty-six percent (58.2% in lung cancer and 75.5% in breast cancer [P < 0.001]) believe in improved disease-free survival. Although 82% believe that systemic therapy can prevent symptoms and prolong life in advanced asymptomatic disease, half (lung cancer 50.8%, breast cancer 53.1% [P = 0.5]) of the PCPs would refer symptomatic patients with advanced disease to palliative care before referral to an oncologist. The type and stage of cancer, as well as the patient's desire or reluctance to be referred to an oncologist were rated by PCPs as the most important reasons to refer patients to an oncologist (P < 0.0001). CONCLUSIONS: Although a majority of PCPs in academia and the community acknowledge the positive effect of chemotherapy, the benefit of systemic therapy for early-stage lung cancer is less appreciated as compared with breast cancer. Patients' preferences influence PCPs significantly in the decision to refer patients to an oncologist.


Assuntos
Antineoplásicos/uso terapêutico , Atitude do Pessoal de Saúde , Neoplasias da Mama/tratamento farmacológico , Competência Clínica , Neoplasias Pulmonares/tratamento farmacológico , Atenção Primária à Saúde , Centros Médicos Acadêmicos , Adulto , Neoplasias da Mama/terapia , Centros Comunitários de Saúde , Medicina de Família e Comunidade , Feminino , Pesquisas sobre Atenção à Saúde , Humanos , Medicina Interna , Kentucky , Neoplasias Pulmonares/terapia , Masculino , Oncologia , Pessoa de Meia-Idade , Encaminhamento e Consulta
10.
Int J Biometeorol ; 58(5): 741-52, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23525898

RESUMO

A study was conducted using 360 broiler chickens to evaluate the effects of dietary vitamin E (0, 125 and 250 mg/kg), selenium (Se, 0, 0.5 and 1 mg/kg), or their different combinations on immune response and blood biological parameters of broilers raised under either thermoneutral (TN, 23.9 °C constant) or heat stress (HS, 23.9 to 37 °C cycling) conditions. Humoral immunity was assessed by intravenous injection of 7% sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Heterophil to lymphocyte (H/L) ratio also determined as an indicator of stress. Furthermore, at the end of the experiment, birds were bled for determination of some biological parameters. There was a significant reduction in body weight and feed intake, but the feed conversion ratio increased when the birds were exposed to HS (P<0.05). Body weight and feed intake were not influenced significantly by dietary vitamin E and Se (P>0.05), whereas feed conversion was improved significantly by 125 mg/kg vitamin E (P<0.05). The liver and lymphoid organ weights as well as IgM and IgG, antibody titers for primary and secondary antibody responses to SRBC were reduced significantly under HS (P<0.05). Heat stress also resulted in a significant increase in H/L ratio (P<0.05). Dietary vitamin E resulted in improvement of primary and secondary antibody responses both in TN and HS broilers (P<0.05). The HS birds also showed an improved antibody titer in secondary response with high concentration of Se (P<0.05). Vitamin E and Se had interactive effects on anti-SRBC titers; however, no consistent differences were found between dietary levels during the study. The H/L ratio decreased by feeding vitamin E at both levels either under HS or TN conditions (P < 0.05). The serum concentrations of glucose, triglycerides, total cholesterol, and LDL-cholesterol were increased but serum HDL-cholesterol decreased in HS broilers (P<0.05).


Assuntos
Antioxidantes/farmacologia , Transtornos de Estresse por Calor/sangue , Transtornos de Estresse por Calor/imunologia , Selênio/farmacologia , Vitamina E/farmacologia , Ração Animal , Animais , Anticorpos/imunologia , Glicemia/análise , Galinhas , Dieta , Ingestão de Alimentos , Eritrócitos/imunologia , Transtornos de Estresse por Calor/veterinária , Imunidade Humoral/efeitos dos fármacos , Lipídeos/sangue , Fígado/efeitos dos fármacos , Fígado/crescimento & desenvolvimento , Linfócitos/imunologia , Tamanho do Órgão/efeitos dos fármacos , Ovinos , Baço/efeitos dos fármacos , Baço/crescimento & desenvolvimento , Temperatura , Timo/efeitos dos fármacos , Timo/crescimento & desenvolvimento
11.
Theriogenology ; 80(3): 193-8, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23726295

RESUMO

The objectives of this study were to compare the following: (1) hatchability, (2) chicks' body and internal organs weights, and (3) plasma gonadotropin levels of hatchlings after in ovo administration of royal jelly (RJ) on Day 7 of incubation. Fertile eggs (n = 270) were injected into the air sac or yolk sac with 0.5 mL normal saline solution consisting of four formulations (normal saline solution with antibiotics, ultrafiltrate RJ, pure RJ, and RJ with antibiotics). The eggs were randomly divided into nine groups of 30 eggs each: (i) C: the control eggs received no injection, (ii) ASA: air sac-injected eggs received normal saline solution with antibiotics, (iii) ARJ: air sac-injected eggs received pure RJ, (iv) ARJA: air sac-injected eggs received RJ with antibiotics, (v) ARJF: air sac-injected eggs received RJ ultrafiltrate solution, (vi) YSA: yolk sac-injected eggs received normal saline solution with antibiotics, and (vii) YRJ: yolk sac-injected eggs received pure RJ, (viii) YRJA: yolk sac-injected eggs received RJ with antibiotics, and (ix) YRJF: yolk sac-injected eggs received ultrafiltrate RJ solution. Hatchability rate was lower in ARJ (46.7%), ARJA (43.3%), ARJF (43.3%), and YRJF (46.7%) groups than in the control (80.0%; P < 0.05). Hatchability rate in ASA (70.0%), YSA (66.7%), YRJ (66.7%), and YRJA (63.3%) groups were comparable to the control (P > 0.05). In ovo injection of RJ and or RJ with antibiotics in both sacs increased chicks' body weight (CWT), heart weight (HWT), and liver weight (LWT) and FSH and LH levels compared with control (P < 0.05). CWT in YRJ (37.02 g), YRJA (37.03 g), ARJ (36.82 g), and ARJA (36.89 g) groups were higher than control (34.9 g; P < 0.05). Similarly, HWT significantly increased in YRJ (0.22 g), YRJA (0.21 g), ARJ (0.20 g), and ARJA (0.20 g) in comparison to control (0.18 g; P < 0.05). In addition, LWT were higher in YRJ (0.83 g), YRJA (0.82g), ARJ (0.81g), and ARJA (0.81g) than control (0.72 g; P < 0.05). Six hours post-hatch, the mean plasma FSH and LH levels in ARJ (1.13 and 2.80 mIu/mL), YRJ (1.32 and 3.36 mIu/mL), ARJA (1.23 and 2.95 mIu/mL), and YRJA (1.31 and 3.28 mIu/mL) groups were higher than in the control (0.56 and 1.48 mIu/mL, P < 0.05). We concluded that in ovo administration of RJ or RJ with antibiotics might be an effective method to increase CWT, chicks' internal organs weights, and LH and FSH secretion rate without deleterious effect on hatchability. However, further research should be conducted to determine the putative endocrine disruptive effects of RJ and its byproducts.


Assuntos
Embrião de Galinha/efeitos dos fármacos , Galinhas/sangue , Ácidos Graxos/administração & dosagem , Ácidos Graxos/farmacologia , Gonadotropinas/sangue , Animais , Embrião de Galinha/crescimento & desenvolvimento , Feminino , Injeções/veterinária
12.
Trop Anim Health Prod ; 44(8): 1865-71, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22528535

RESUMO

The objective of this study was to evaluate if treatment of equine chorionic gonadotrophin (eCG)-superovulated Sanjabi ewes with repeated administration of human chorionic gonadotropin (hCG) would increase the number of normal corpus luteum (CL) and serum progesterone concentrations and decrease the number of persistent follicles. The superovulated ewes were divided into four groups on day 0 (day of sponge removal); the ewes were treated by an intramuscular administration of 500 IU hCG on day 0 (Group I: n = 10), on days 0 and 1 (Group II: n = 10), or on days 0, 1, and 2 (Group III: n = 10) and no treatment for control group (n = 10). Blood samples were collected on days 0, 1, 2, 5, and 8 (day of slaughter), and serum progesterone concentrations were determined. According to progesterone concentrations, 50 (4/8) and 0 % of the ewes underwent premature luteal regression in the control group and the hCG groups, respectively. There were more CLs in Group III than in Group II and the control group. Ewes treated with hCG had a greater number of normal-looking CL. CL diameter was significantly greater in Group II and Group III than other groups. Total CL weight was significantly (P < 0.05) higher in Group III than in Group I and the control group. Number of persistent follicle and persistent follicle diameter were lower in control group compared to the other groups. Eight days after sponge removal, serum progesterone concentration was significantly higher in Group III than in Group I and the control group. The present results indicate that repeated administration of hCG supported CL formation, increased serum progesterone concentration, and prevented premature luteal regression in eCG-superovulated Sanjabi ewes.


Assuntos
Gonadotropina Coriônica/administração & dosagem , Corpo Lúteo/efeitos dos fármacos , Luteólise/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Progesterona/sangue , Carneiro Doméstico/fisiologia , Animais , Corpo Lúteo/citologia , Corpo Lúteo/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Gonadotropinas Equinas/administração & dosagem , Cavalos , Humanos , Irã (Geográfico) , Modelos Logísticos , Folículo Ovariano/citologia , Folículo Ovariano/crescimento & desenvolvimento , Superovulação/efeitos dos fármacos
13.
Asian-Australas J Anim Sci ; 25(4): 447-51, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25049584

RESUMO

The objective of this research was to determine the prevalence, risk factors and consequent effect of dystocia on lactation performance in Holstein dairy cows in Iran. The data set consisted of 55,577 calving records on 30,879 Holstein cows in 30 dairy herds for the period March 2000 to April 2009. Factors affecting dystocia were analyzed using multivariable logistic regression models through the maximum likelihood method in the GENMOD procedure. The effect of dystocia on lactation performance and factors affecting calf birth weight were analyzed using mixed linear model in the MIXED procedure. The average incidence of dystocia was 10.8% and the mean (SD) calf birth weight was 42.13 (5.42) kg. Primiparous cows had calves with lower body weight and were more likely to require assistance at parturition (p<0.05). Female calves had lower body weight, and had a lower odds ratio for dystocia than male calves (p<0.05). Twins had lower birth weight, and had a higher odds ratio for dystocia than singletons (p<0.05). Cows which gave birth to a calf with higher weight at birth experienced more calving difficulty (OR (95% CI) = 1.1(1.08-1.11). Total 305-d milk, fat and protein yield was 135 (23), 3.16 (0.80) and 6.52 (1.01) kg less, in cows that experienced dystocia at calving compared with those that did not (p<0.05).

14.
BMC Biotechnol ; 10: 9, 2010 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-20144189

RESUMO

BACKGROUND: The delivery of DNA into human cells has been the basis of advances in the understanding of gene function and the development of genetic therapies. Numerous chemical and physical approaches have been used to deliver the DNA, but their efficacy has been variable and is highly dependent on the cell type to be transfected. RESULTS: Studies were undertaken to evaluate and compare the transfection efficacy of several chemical reagents to that of the electroporation/nucleofection system using both adherent cells (primary and transformed airway epithelial cells and primary fibroblasts as well as embryonic stem cells) and cells in suspension (primary hematopoietic stem/progenitor cells and lymphoblasts). With the exception of HEK 293 cell transfection, nucleofection proved to be less toxic and more efficient at effectively delivering DNA into the cells as determined by cell proliferation and GFP expression, respectively. Lipofectamine and nucleofection of HEK 293 were essentially equivalent in terms of toxicity and efficiency. Transient transfection efficiency in all the cell systems ranged from 40%-90%, with minimal toxicity and no apparent species specificity. Differences in efficiency and toxicity were cell type/system specific. CONCLUSIONS: In general, the Amaxa electroporation/nucleofection system appears superior to other chemical systems. However, there are cell-type and species specific differences that need to be evaluated empirically to optimize the conditions for transfection efficiency and cell survival.


Assuntos
DNA/química , Transfecção/métodos , Animais , Linhagem Celular , Sobrevivência Celular , Eletroporação , Células-Tronco Embrionárias/metabolismo , Células Epiteliais/metabolismo , Fibroblastos/metabolismo , Humanos , Lipídeos/química , Camundongos , Polietilenoimina/química , Coelhos , Suínos
15.
Oligonucleotides ; 16(3): 213-24, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16978085

RESUMO

An ultimate goal of gene therapy is the development of a means to correct mutant genomic sequences in the cells that give rise to pathology. A number of oligonucleotide-based gene-targeting strategies have been developed to achieve this goal. One approach, small fragment homologous replacement (SFHR), has previously demonstrated disease-specific genotypic and phenotypic modification after introduction of small DNA fragments (SDFs) into somatic cells. To validate whether the gene responsible for sickle cell anemia (beta-globin) can be modified by SFHR, a series of studies were undertaken to introduce sickle globin sequences at the appropriate locus of human hematopoietic stem/progenitor cells (HSPCs). The characteristic A two head right arrow T transversion in codon 6 of the beta-globin gene was indicated by restriction fragment length polymorphic (RFLP) analysis of polymerase chain reaction (PCR) products generated by amplification of DNA and RNA. At the time of harvest, it was determined that the cells generally contained

Assuntos
Marcação de Genes , Globinas/genética , Células-Tronco Hematopoéticas/metabolismo , Oligodesoxirribonucleotídeos/metabolismo , Células Cultivadas , Sangue Fetal/citologia , Genoma Humano , Humanos , Oligodesoxirribonucleotídeos/genética
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