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1.
Chemosphere ; 362: 142433, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38815812

RESUMO

Pesticides are becoming more prevalent in agriculture to protect crops and increase crop yields. However, nearly all pesticides used for this purpose reach non-target crops and remain as residues for extended periods. Contamination of soil by widespread pesticide use, as well as its toxicity to humans and other living organisms, is a global concern. This has prompted us to find solutions and develop alternative remediation technologies for sustainable management. This article reviews recent technological developments for remediating pesticides from contaminated soil, focusing on the following major points: (1) The application of various pesticide types and their properties, the sources of pesticides related to soil pollution, their transport and distribution, their fate, the impact on soil and human health, and the extrinsic and intrinsic factors that affect the remediation process are the main points of focus. (2) Sustainable pesticide degradation mechanisms and various emerging nano- and bioelectrochemical soil remediation technologies. (3) The feasible and long-term sustainable research and development approaches that are required for on-site pesticide removal from soils, as well as prospects for applying them directly in agricultural fields. In this critical analysis, we found that bioremediation technology has the potential for up to 90% pesticide removal from the soil. The complete removal of pesticides through a single biological treatment approach is still a challenging task; however, the combination of electrochemical oxidation and bioelectrochemical system approaches can achieve the complete removal of pesticides from soil. Further research is required to remove pesticides directly from soils in agricultural fields on a large-scale.


Assuntos
Agricultura , Biodegradação Ambiental , Recuperação e Remediação Ambiental , Praguicidas , Poluentes do Solo , Solo , Poluentes do Solo/análise , Praguicidas/análise , Praguicidas/química , Agricultura/métodos , Recuperação e Remediação Ambiental/métodos , Solo/química , Humanos
2.
Food Chem ; 443: 138520, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38290296

RESUMO

Present study reports fabrication of a low cost and eco-friendly formaldehyde nanosensor based on green magnetite nanoparticles synthesized using Mango (Mangifera indica L.) tree leaves extract. The formaldehyde is found in air, water and food. When inhaled or consumed formaldehyde has carcinogenic effects on human health. In this study the cyclic voltammetry technique was used to characterize the performance of the nanosensor. The green nanosensor fabricated in this study, to detect formaldehyde, demonstrated good sensitivity (193.4 µA mg-1 Lcm-2) in linearity range 0.03-0.5 mg/L with low threshold detection limit (0.05 mg/L). The green nanosensor also showed shelf life of four weeks without considerable change in the initial peak oxidation current. The real sample analysis was performed in various fruits and vegetables (Litchi chinensis, Syzygium cumini, Solanum lycopersicum and Cucumis sativus). The recovery rates were more than 93 % in sample extracts for formaldehyde detection. The comparison of the nanosensor for detection of formaldehyde with the colorimetric sensor revealed that the green nanosensor reproducibility (RSD = 1.8 %) is better than colorimetric sensor (RSD = 3.23 %). The results from the comparative studies of green nanosensor with colorimetric sensor established the potential of the green nanosensor as a forefront technology for futuristic smart detection of formaldehyde.


Assuntos
Frutas , Verduras , Humanos , Frutas/química , Reprodutibilidade dos Testes , Colorimetria/métodos , Formaldeído/análise
3.
Environ Toxicol Pharmacol ; 88: 103740, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34506906

RESUMO

The accumulation of heavy metals (HMs) in soil is presently a significant threat to the environment. The Cu, Mn, Zn, Cd, Pb, Ni, and Co concentrations were assessed in the agricultural soil samples. The results of various contamination indices including contamination factor, geo-accumulation indices, and ecological risk indices revealed that Cd is responsible for moderate to high contamination of soil. The multivariate statistical analyses including PCA, HCA, and correlation matrix suggested the mixed origin of HMs in the soil. Ingestion was found to be a primary route of exposure while dermal and inhalation exposure was negligible. Overall, the non-carcinogenic health risks were well within the safe limit to human health. However, healthwise, children were likely to be at greater risk compared with adults, due to their generally increased exposure to toxic agents through hand/mouth ingestion. Moreover, no carcinogenic risks were determined through the inhalation exposure of Cd, Ni, and Co.


Assuntos
Metais Pesados/análise , Poluentes do Solo/análise , Adulto , Agricultura , Criança , Cidades , Monitoramento Ambiental , Humanos , Índia , Neoplasias , Medição de Risco
4.
Trop Anim Health Prod ; 53(2): 220, 2021 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-33751251

RESUMO

Limited information exists on the application of human chorionic gonadotropin (hCG) post insemination in the Indian crossbred dairy cows. This study aimed to evaluate the effect of four hCG administration protocols on luteal characteristics and pregnancy outcomes following artificial insemination (AI). Using block randomization, 50 healthy lactating crossbred cows were allocated in equal numbers (n = 10 cows per group) to receive either 5 mL normal saline (control) or 1500 IU hCG on the day of AI (hGG-0), day 7 post AI (hCG-7), day 14 post AI (hCG-14), or days 0, 7, and 14 post AI (hCG-0,7,14). All cows were scanned using sequential transrectal ultrasound examinations to evaluate primary luteal parameters, development of accessory corpora lutea, and pregnancy. Serial blood samples were collected to measure plasma progesterone concentrations. Data were analyzed using repeated measures ANOVA and Fisher's exact tests. The mean primary luteal area, total luteal area, and total luteal diameter values were significantly greater in the hCG-treated cows. Compared to the control, the hCG-14 group had a significantly higher percentage of cows with an accessory corpora luteum. However, there were no significant differences in the mean progesterone concentrations or the first service conception rates between any of the groups. Overall, the results of this study indicate that while hCG administration post AI in healthy Indian crossbred cows may enhance primary luteal dimensions or induce the formation of accessory corpora lutea, it does not appear to have any beneficial effect on luteal function or pregnancy.


Assuntos
Corpo Lúteo , Lactação , Animais , Bovinos , Gonadotropina Coriônica/farmacologia , Protocolos Clínicos , Sincronização do Estro , Feminino , Hormônio Liberador de Gonadotropina , Inseminação Artificial/veterinária , Plasma , Gravidez , Progesterona
5.
Indian J Palliat Care ; 25(4): 567-574, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31673214

RESUMO

CONTEXT: Inadequate training of medical practitioners is a key factor responsible for inappropriate use of opioid analgesics. AIMS: We assessed the current knowledge, attitude, prescribing practices, and barriers perceived by the Indian medical practitioners in three tertiary care hospitals toward the use of opioid analgesics. SUBJECTS AND METHODS: Web-based survey of registered medical practitioner employed at three chosen tertiary health care institutions in New Delhi. STATISTICAL ANALYSIS USED: Descriptive analysis of survey responses was carried out. Comparative analysis was done using Chi-square test, independent samples t-test, and Pearson correlation coefficient. RESULTS: The response rate was 10.4% (n = 308). Two-thirds of the participants (61.7%) had never received formal pain management training, and 86.7% participants would like further training. Most participants (71.1%) agreed that opioids should be prescribed in cancer pain, while 26.3% agreed that opioids should be prescribed in noncancer pain. Half of the participants agreed that SOS (if necessary) dosing schedule (48.4%), low dosage (61.7%), and short duration of use (51.4%) could decrease the harmful effect of opioids. Lack of information about opioid-related policies and addiction potential were identified as the most common barriers to prescribing opioids. Those seeing more patients with chronic noncancer pain come across opioid misuse and diversion more often (P = 0.02). Those who understood addiction were more likely to agree that patients of chronic cancer pain with substance use disorders should be prescribed opioid analgesics (P < 0.01). CONCLUSIONS: Indian medical practitioners felt the need for formal pain management training. There is a lack of consensus on how to manage the pain using opioid analgesics. Tough regulations on medical and scientific use of opioids are the most commonly reported barrier to prescribing them.

6.
Reumatol Clin ; 13(4): 224-226, 2017.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-27174399

RESUMO

A 24-year-old female presented with catatonia and symptoms suggestive of Depressive Disorder. She also gave history of undocumented low grade irregular fever. The patient was worked up to rule out any organic cause or psychiatric illness. However, further investigations revealed immunological profile diagnostic of Systemic Lupus Erythematosus (SLE) with CNS involvement (CNS lupus). The diagnosis of SLE in this patient presenting with catatonia was of practical importance because catatonia as one of the manifestations of SLE or as a standalone presenting symptom is extremely rare. Hence, clinicians should be aware of this rarity so that diagnosis of Neuropsychiatric SLE (NPSLE) or catatonia as a presenting feature of SLE is never missed.


Assuntos
Catatonia/etiologia , Lúpus Eritematoso Sistêmico/diagnóstico , Feminino , Humanos , Lúpus Eritematoso Sistêmico/complicações , Adulto Jovem
7.
J Ultrasound ; 18(1): 67-70, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25767638

RESUMO

Several studies in dairy cattle have investigated the final outcome of different treatment regimens in follicular cyst condition. However, sequential monitoring of the response of follicular cysts to these treatments is rather scanty. In this paper, we present the response of a large follicular cyst in a pluriparous crossbred dairy cow with prolonged conception failure to human chorionic gonadotropin, hCG (3,000 IU; day 0) and cloprostenol (500 µg; day 9) treatment. Using transrectal ultrasonography (USG), reproductive tract was imaged daily beginning day 0 until day 11. The follicular cyst showed a consistent regression to a very small anechoic area on day 7 and was undetectable thereafter. Concurrently, there was development of a new dominant follicle that was first detected on day 4 and showed progressive growth to preovulatory stage. The cow was inseminated and ovulation occurred, as diagnosed by the presence of a corpus luteum (CL) 7 days later, but conception did not occur. The animal was re-inseminated after estrus detection in the estrous cycle that immediately followed. Pregnancy diagnosis was performed on 30 and 60 days post-insemination (DPI) and the cow was confirmed to be pregnant. This paper underscores the importance of diagnostic ultrasound in veterinary medicine, especially in the management of reproductive problems.

8.
Vaccine ; 32(1): 11-8, 2013 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-24211168

RESUMO

Inhibin and follistatin are known to reduce fecundity by inhibiting the actions of activin and FSH. Thus, the immunoneutralization of these hormones is a rational proposal for augmenting reproductive performance. The present study describes a comprehensive computational methodology comprising of a consensus approach of several B- and Th-cell epitope prediction tools for the identification of epitopic regions within the structure of these hormones that can be incorporated into a poly-epitope fecundity vaccine. The proposed peptide (RGD-WSPAALRLLQRPPEEPA-KK-YSFPISSILE) should be effective in multiple animal species, generating good immunological memory.


Assuntos
Epitopos/imunologia , Fertilidade/imunologia , Gado/imunologia , Vacinas/imunologia , Sequência de Aminoácidos , Animais , Bovinos , Galinhas , Mapeamento de Epitopos/métodos , Epitopos/química , Epitopos de Linfócito B/química , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/química , Epitopos de Linfócito T/imunologia , Folistatina/química , Folistatina/imunologia , Cavalos , Inibinas/química , Inibinas/imunologia , Camundongos , Modelos Moleculares , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/imunologia , Conformação Proteica , Ratos , Ovinos , Sus scrofa , Vacinas de Subunidades Antigênicas/química , Vacinas de Subunidades Antigênicas/imunologia
9.
Vet Res Forum ; 4(2): 85-9, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-25653777

RESUMO

Four crossbred cows with mummified fetus were utilized for the study. The cows were subjected to gynecological examination and based on the findings the cases were diagnosed as mummified fetus. The cows were treated with 2 mg estradiol valerate and 500 µg cloprostenol and were examined every 12 hr after 24 hr of the treatment for cervical dilatation and other signs related to fetal expulsion. The time duration between treatment and starting of cervical dilatation ranged from 48 to 58h (53.00 ± 2.08 hr). Complete dilatation of cervix was observed after 70.00 ± 2.94 hr post treatment (Range = 64-76 hr). The mean fetal crown-rump length (CRL) was 31.5 cm, which ranged from 27.5 to 38 cm. The number of cotyledons in pregnant horn also showed wide variation (Range 24-38 numbers) with mean ± SE of 30.3 ± 3.07 numbers. In the placenta of three animals irregular shaped large adventitious cotyledons were observed in the inter-cotyledonary areas. Out of the four animals treated, three animals were conceived within three estrous cycles and one animal had cystic ovary in the next cycle and was not conceived even after four cycles. It was concluded that the estradiol and prostaglandin F2α (PGF2α) combination therapy was effective for expulsion of mummified fetus in crossbred cows without affecting much on future fertility.

10.
Trop Anim Health Prod ; 44(7): 1697-702, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22476734

RESUMO

The present study was undertaken to evaluate the effect of L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor on serum nitric oxide, progesterone, estradiol profiles and estrous cycle length in buffaloes. Murrah buffaloes (n = 16) exhibiting regular estrous cycles were randomly allocated to two groups of eight animals. In the treatment group, buffaloes were administered 400 mg/h L-NAME over 2 h (total dose = 800 mg) via the coccygeal artery and the aorta abdominalis on day 15 of the estrous cycle. In the control group, normal saline was infused on the same day of the cycle by the same route. Blood samples were collected every 4 h on days 15 and 16, and once daily from days 17 to 21 of the estrous cycle for the assay of progesterone, estradiol and nitric oxide. L-NAME treatment significantly (p < 0.05) reduced serum nitric oxide concentration from 4 h of day 15 until day 20 of the cycle. Serum progesterone concentration increased significantly (p < 0.05) between 0 and 20 h post treatment on day 15. The estrous cycle length was 19.8 ± 0.36 and 23.6 ± 0.17 days for control and treated group buffalo (p < 0.05), respectively. It was concluded that treatment of buffalo with L-NAME in the late luteal phase of the estrous cycle inhibited serum nitric oxide concentration resulting in increased progesterone production and extension of the effective life of the corpus luteum, thus prolonging the estrous cycle length.


Assuntos
Búfalos/fisiologia , Corpo Lúteo/efeitos dos fármacos , Inibidores Enzimáticos/administração & dosagem , Estradiol/sangue , Ciclo Estral/efeitos dos fármacos , NG-Nitroarginina Metil Éster/administração & dosagem , Óxido Nítrico/sangue , Progesterona/sangue , Animais , Dinoprosta/administração & dosagem , Feminino , Infusões Intra-Arteriais/veterinária , Fatores de Tempo
11.
Appl Biochem Biotechnol ; 166(7): 1814-30, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22350939

RESUMO

The present study characterized the glycoproteins synthesized by buffalo oviduct. Scanning electron microscopy analyses of the ampullary and isthmic segments of cyclic and acyclic buffaloes showed ultrastructural variations in ciliated and nonciliated cells. Mucosal proteins were extracted by scrapping of different segments of oviduct and, after centrifugation, the remainder tissues were subjected to establish primary cell culture system of oviduct epithelial cells and conditioned media were prepared. Time- and concentration-dependent effects of trypsinization on the establishment of primary monolayer culture showed that 0.25% trypsin for 1-2 min at 37 °C were the optimal conditions. Total protein content in oviductal tissues and conditioned media was quantified and analyzed by SDS-PAGE which showed marked variation in different segments of the oviduct. Western blot analysis revealed five major oviduct-specific glycoproteins (OGPs) in cyclic oviduct (ampulla and isthmus) with Mw 180, 95, 75, 66 and 35 kDa in the oviduct extract and two glycoproteins with Mw 95 and 66 kDa in conditioned media. However, in acyclic oviduct (ampulla and isthmus), three glycoproteins were immunostained with Mw 180, 95 and 66 kDa in the oviduct extract and one glycoprotein with Mw 66 kDa in conditioned media. Indirect Enzyme-Linked Immuno Sorbent Assay (ELISA) results showed significant differences of OGPs in different segments of cyclic and acyclic buffaloes and, thus, indicative of segmental variation in the synthesis and secretion of glycoproteins. Oviductal extract secretes more amounts of OGPs as compared to the conditioned medium. The role of these OGPs may be defined and exploited for influencing the fertilization process and/or subsequent embryonic development.


Assuntos
Células Epiteliais/citologia , Glicoproteínas/isolamento & purificação , Mucosa/citologia , Oviductos/citologia , Animais , Western Blotting , Búfalos/fisiologia , Sobrevivência Celular , Microambiente Celular , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/química , Células Epiteliais/fisiologia , Feminino , Fertilidade/fisiologia , Glicoproteínas/química , Ciclo Menstrual/fisiologia , Microscopia Eletrônica de Varredura , Peso Molecular , Mucosa/fisiologia , Oviductos/fisiologia , Gravidez , Cultura Primária de Células , Tripsina/metabolismo
12.
J Mol Neurosci ; 43(3): 428-42, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20953735

RESUMO

Preconditioning and postconditioning are mild ischemic exposures before or after severe injurious ischemia, respectively, that elicit endogenous neuroprotective responses. Molecular mechanisms of neuroprotection through preconditioning and postconditioning are not completely understood. Here we optimized the in vitro oxygen and glucose deprivation (OGD) models of preconditioning and postconditioning in primary cortical neuron cultures that allow the studies of the corresponding molecular mechanisms of neuroprotection. We found that the cortical cells preconditioned with a single 45-min OGD treatment administered 24 h prior to injurious 2 h OGD were robustly protected after both 3 h and 16 h of reperfusion. For the postconditioning treatment, we found that three cycles of 15 min OGD followed by 15 min reperfusion, applied immediately after injurious 2 h OGD and prior to complete reperfusion, resulted in effective neuroprotection at both 3 h and 16 h of reperfusion. Using real-time RT-PCR arrays focused on genes of the apoptosis and PI3K-Akt pathways, we found that injurious OGD mainly induced apoptosis-related and repressed PI3K-Akt pathway-related genes after either 3 h or 16 h of reperfusion. Preconditioning treatment resulted in the activation of both pro-survival and anti-apoptotic pathways after 3 h of reperfusion and mainly anti-apoptotic pathway after 16 h of reperfusion. In contrast, the activation of PI3K-Akt pathway mainly contributed to the neuroprotective effect by the postconditioning treatment after 3 h of reperfusion, but differential gene expression likely contributed minimally, if at all, to the neuroprotection observed after 16 h of reperfusion. Among the novel markers of neuroprotection, Nol3 gene upregulation was observed after 3 h of reperfusion following either preconditioning or postconditioning treatments and after 16 h of reperfusion following preconditioning treatment.


Assuntos
Apoptose/fisiologia , Precondicionamento Isquêmico , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Animais , Biomarcadores/metabolismo , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Células Cultivadas , Córtex Cerebral/citologia , Perfilação da Expressão Gênica , Glucose/metabolismo , Neurônios/citologia , Neurônios/fisiologia , Oxigênio/metabolismo , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Ratos Wistar
13.
Invest Ophthalmol Vis Sci ; 51(2): 1151-63, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19797223

RESUMO

PURPOSE: Age-related macular degeneration (AMD) is a common cause of irreversible vision loss in the elderly. The hypothesis was that in vitro stimulation of RPE cells with Abeta(1-40), a constituent of drusen, promotes changes in gene expression and cellular pathways associated with the pathogenesis of AMD, including oxidative stress, inflammation, and angiogenesis. METHODS: Confluent human RPE cells were stimulated with Abeta(1-40), or the reverse peptide Abeta(40-1), and genome wide changes in gene expression were studied with gene microarrays. Selected genes were verified by qRT-PCR and ELISA. Pathway analysis with gene set enrichment analysis (GSEA) and ingenuity revealed top functional pathways in RPE after Abeta(1-40) stimulation. RESULTS: RPE cells stimulated with Abeta(1-40) (0.3 microM) for 24 hours resulted in 63 upregulated and 22 downregulated previously known genes. The upregulated genes were predominantly in inflammatory and immune response categories, but other categories were also represented, including apoptosis, cell signaling, cell proliferation, and signal transduction. Categories of downregulated genes included immune response, transporters, metabolic functions and transcription factors. ELISA confirmed that secreted levels of IL-8 were two times higher than control levels. GSEA and ingenuity analysis confirmed that the top affected pathways in RPE cells after Abeta(1-40) stimulation were inflammation and immune response related. Surprisingly, few angiogenic pathways were activated at the doses and exposure times studied. CONCLUSIONS: Abeta(1-40) promotes RPE gene expression changes in pathways associated with immune response, inflammation, and cytokine and interferon signaling pathways. Results may relate to in vivo mechanisms associated with the pathogenesis of AMD.


Assuntos
Peptídeos beta-Amiloides/farmacologia , Indutores da Angiogênese/metabolismo , Citocinas/genética , Proteínas do Olho/genética , Regulação da Expressão Gênica/fisiologia , Fragmentos de Peptídeos/farmacologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Proteínas do Olho/metabolismo , Perfilação da Expressão Gênica , Humanos , Immunoblotting , Microscopia de Força Atômica , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Anim Reprod Sci ; 113(1-4): 44-50, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18620823

RESUMO

This study examined the effect of including one of two growth factors (100 ng/ml IGF-1 or 20 ng/ml EGF) in combination with one of two antioxidants (50 microM cysteamine or 50 microM beta-mercaptoethanol) in maturation, fertilization and subsequent development of buffalo oocytes. The oocytes were recovered by in vivo ovum pick-up technique from six Murrah buffalo heifers twice a week over a period of 16 weeks. Immediately after ovum pick-up oocytes recovered from six donors were allocated randomly to five different maturation treatments. The control treatment was the basic maturation medium (MM; TCM-199 supplemented with 10% FBS, 10 IU/ml LH, 0.5 microg/ml FSH, 1 microg/ml estradiol-17beta and 50 microg/ml gentamicin). The other four treatments consisted of the control maturation medium (MM) plus one combination of a growth factor and an antioxidant viz. IGF-1+cysteamine; IGF-1+beta-ME; EGF+cysteamine or EGF+beta-ME. The total number of oocytes assigned to each maturation treatment ranged from 31 to 66. After maturation in different maturation medium, media used for in vitro fertilization and subsequent development of embryo was same for all groups. Data were analysed using Chi-square test. The maturation rate observed for the growth factor plus antioxidant treatments was similar to that for the control (90.4%). The highest cleavage rate recorded in the IGF-1+cysteamine treatment (71.9%) which was significantly higher (P<0.05) than the IGF-1+beta-ME (45.2%) and EGF+beta-ME (46.4%) treatments, but not significantly differ from the control (63.8%) and EGF+cysteamine treatment (60.7%). The proportion of cleaved oocytes those developed to blastocyst stage was significantly higher in the IGF-1+cysteamine treatment (52.2%; P<0.05) than in the control (23.3%), the EGF+cysteamine (13.5%) or the EGF+beta-ME (7.7%) treatments, but did not differ significantly from the IGF-1+beta-ME (28.6%) treatment. Following non-surgical transfer of 15 embryos to 14 synchronized recipients, four became pregnant and only one recipient sustained the pregnancy as long as 4.5 months when spontaneous abortion occurred. It was concluded that supplementing the maturation medium with IGF-1+cysteamine improved the production of buffalo embryos significantly in vitro culture.


Assuntos
Antioxidantes/farmacologia , Búfalos , Meios de Cultura/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Oócitos/efeitos dos fármacos , Oogênese/efeitos dos fármacos , Animais , Búfalos/fisiologia , Meios de Cultura/química , Técnicas de Cultura Embrionária/métodos , Feminino , Fertilização in vitro/veterinária , Recuperação de Oócitos/veterinária , Oócitos/fisiologia , Gravidez , Taxa de Gravidez
15.
Invest Ophthalmol Vis Sci ; 46(4): 1508-15, 2005 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15790923

RESUMO

PURPOSE: To examine the effect of loss of cell-cell contacts on the gene expression of vascular endothelial growth factor (VEGF) and other factors in primary culture of human retinal pigment epithelial (RPE) cells with real-time reverse transcription-PCR. METHODS: The dissociation of postconfluent RPE cells was induced by calcium chelation, low-calcium medium, anti-E-cadherin, and anti-N-cadherin antibodies. Total RNA was isolated from the cultured RPE cells and reverse transcribed to cDNA. VEGF was quantified by real-time PCR with a fluorescence detector. VEGF isoforms were differentially measured by specific exon-spanning primers. Besides VEGF, the gene expression levels of some other growth factors were also examined in calcium-mediated dissociation. RESULTS: Disruption of cell-cell contacts of RPE cells was induced by calcium chelation and low-calcium medium, but not by anti-E-cadherin and anti-N-cadherin antibodies. Calcium-mediated dissociation of RPE cells significantly increased the gene expression levels of VEGF. The mRNA levels of VEGF increased by 6.3-fold on treatment with EGTA and by 4.7-fold in the low-calcium medium at 6 hours. Splice variants of VEGF showed the differential pattern of gene expression. Whereas the expression of VEGF(121) and VEGF(165) was upregulated on calcium-induced dissociation of RPE cells, that of VEGF(145) and VEGF(189) was unchanged. VEGF(206) was not detected. On calcium-induced dissociation, bFGF, IL-6, matrix metalloproteinase (MMP)-1, and placental growth factor (PlGF) were upregulated, whereas acidic (a)FGF and pigment epithelium-derived factor (PEDF) were both downregulated. CONCLUSIONS: The results show that loss of intercellular contacts promotes increased gene expression of VEGF and other angiogenic factors in human RPE cells.


Assuntos
Cálcio/farmacologia , Expressão Gênica/efeitos dos fármacos , Epitélio Pigmentado Ocular/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genética , Actinas/metabolismo , Caderinas/metabolismo , Adesão Celular/efeitos dos fármacos , Comunicação Celular/efeitos dos fármacos , Células Cultivadas , Quelantes/farmacologia , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Epitélio Pigmentado Ocular/metabolismo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/metabolismo
16.
Exp Eye Res ; 79(2): 209-21, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15325568

RESUMO

We examined the gene expression profiles of retinal pigment epithelial (RPE) cells which were aged in vitro by repeated passage. RPE cells from human eyes were cultured to passage 3-5 (early passage) or 19-21 (late passage) and used to study gene expression profiles by cDNA microarray. Results from microarray analysis were further confirmed by real-time PCR. Microarray analysis showed gene expression changes among 588 known genes. The expression levels of 15 genes (2.6%) increased in late passage RPE cells, while 43 genes (7.3%) decreased using a two-fold criterion. These differentially expressed genes encompassed many functional classes. A small number of stress genes, such as clusterin, replication protein A and Ku80, were up-regulated. The down-regulated genes included many enzymes of energy and biomolecule metabolism as well as cell cycle proteins and cell adhesion proteins. Results from real-time PCR were generally consistent with microarray findings. The expression levels of the examined angiogenic factors were either unchanged or down-regulated. Comparing early (p=3-5) and late (p=9-12) passage RPE cells, several categories of differentially expressed genes were identified. However, there was no enhanced expression of known angiogenic factors.


Assuntos
Senescência Celular/genética , Regulação da Expressão Gênica , Degeneração Macular/metabolismo , Epitélio Pigmentado Ocular/citologia , Indutores da Angiogênese/metabolismo , Células Cultivadas , Perfilação da Expressão Gênica/métodos , Humanos , Degeneração Macular/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Epitélio Pigmentado Ocular/metabolismo , Reação em Cadeia da Polimerase/métodos , beta-Galactosidase/metabolismo
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