RESUMO
Allergic conjunctivitis (AC) is the most common form of allergic eye disease and an increasingly prevalent condition. Topical eye drop treatments are the usual approach for managing AC, although their impact on the ocular surface is not frequently investigated. The aim of this study was to perform a comparative physicochemical characterization, and in vitro biological evaluations in primary conjunctival and corneal epithelial cells of the new multidose preservative-free bilastine 0.6% and main commercially available eye drops. MTT assay was used to measure cell viability; oxidative stress was analyzed with a ROS-sensitive probe; and apoptosis was evaluated monitoring caspase 3/7 activation. Differences in pH value, osmolarity, viscosity and phosphate levels were identified. Among all formulations, bilastine exhibited pH, osmolarity and viscosity values closer to tear film (7.4, 300 mOsm/l and ~ 1.5-10 mPa·s, respectively), and was the only phosphates-free solution. Single-dose ketotifen did not induce ROS production, and single-dose azelastine and bilastine only induced a mild increase. Bilastine and single-dose ketotifen and azelastine showed high survival rates attributable to the absence of preservative in its formulation, not inducing caspase-3/7-mediated apoptosis after 24 h. Our findings support the use of the new bilastine 0.6% for treating patients with AC to preserve and maintain the integrity of the ocular surface.
Assuntos
Apoptose , Benzimidazóis , Caspase 3 , Sobrevivência Celular , Soluções Oftálmicas , Conservantes Farmacêuticos , Soluções Oftálmicas/farmacologia , Humanos , Conservantes Farmacêuticos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Benzimidazóis/farmacologia , Benzimidazóis/química , Caspase 3/metabolismo , Apoptose/efeitos dos fármacos , Piperidinas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Túnica Conjuntiva/efeitos dos fármacos , Túnica Conjuntiva/metabolismo , Túnica Conjuntiva/patologia , Caspase 7/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Conjuntivite Alérgica/tratamento farmacológico , Conjuntivite Alérgica/patologia , Conjuntivite Alérgica/metabolismo , Ftalazinas/farmacologia , Concentração Osmolar , Epitélio Corneano/efeitos dos fármacos , Epitélio Corneano/metabolismo , Células Cultivadas , ViscosidadeRESUMO
Enzootic bovine leukosis (EBL) is a chronic infectious disease caused by the bovine leukosis virus (BLV), a Deltaretrovirus. Bovine viral diarrhea (BVD) is an infectious disease caused by a pestivirus. Bovine neosporosis is caused by the obligate intracellular parasite Neospora caninum (Nc). These pathogens can have horizontal (postnatal) or vertical (transplacental) transmissions and affect the productive and reproductive performance of infected bovines. This work aimed to detect BLV, BVD, and Nc seroprevalence in specialized dairy cattle from the north, east, and Aburrá Valley regions of the Department of Antioquia, the highest in milk production regions in Antioquia. A total of 599 blood samples, obtained from 53 specialized dairy cattle herds, were evaluated by the ELISA test. The results revealed a seroprevalence of 41.13% for BLV (242/599), 28.48% (163/599) for Nc, and 22.7% (132/599) for BVD. Regarding the regional seroprevalence evaluation, BLV was found in 47.02% of the samples from the east, 36.87% from the north, and 46.02% from the Aburrá Valley. Nc was found in 31.03% of the samples from the east, 24.26% from the north, and 36.63% from Aburrá Valley. BVD was found in 21.62% of the samples from the east, 25.03% from the north region, and 10.39% of the samples from the Aburrá Valley. It is highlighted by these results that the north region, with the highest milk production in Antioquia, had the lowest BLV and Nc seroprevalences but the highest seroprevalence of BVD. BLV has increased in Antioquia in recent years, and as an immunosuppressive infection, opportunities for other pathogens are created by it. A significant statistical difference was found in the average prevalence of the pathogens according to the municipality, cattle breed, and region of origin of the sample. The seroprevalence of these pathogens in specialized dairy herds in Antioquia can be classified as medium-low. However, it is recommended that biosecurity practices should be maximized to avoid the spread of these pathogens due to the variability detected in the region, municipality, breed group, and herd age. The rapid and efficient diagnosis of these three pathogens through reliable methodologies will allow for the control of dissemination in dairy herds.
Assuntos
Doenças dos Bovinos , Doenças Transmissíveis , Leucose Enzoótica Bovina , Vírus da Leucemia Bovina , Neospora , Animais , Bovinos , Colômbia , Leucose Enzoótica Bovina/epidemiologia , Estudos Soroepidemiológicos , Doenças Transmissíveis/veterinária , Diarreia/veterináriaRESUMO
In diabetes, chronic hyperglycemia, dyslipidemia, inflammation and oxidative stress contribute to the progression of macro/microvascular complications. Recently, benefits of the use of flavonoids in these conditions have been established. This study investigates, in two different mouse models of diabetes, the vasculoprotective effects of the synthetic flavonoid hidrosmin on endothelial dysfunction and atherogenesis. In a type 2 diabetes model of leptin-receptor-deficient (db/db) mice, orally administered hidrosmin (600 mg/kg/day) for 16 weeks markedly improved vascular function in aorta and mesenteric arteries without affecting vascular structural properties, as assessed by wire and pressure myography. In streptozotocin-induced type 1 diabetic apolipoprotein E-deficient mice, hidrosmin treatment for 7 weeks reduced atherosclerotic plaque size and lipid content; increased markers of plaque stability; and decreased markers of inflammation, senescence and oxidative stress in aorta. Hidrosmin showed cardiovascular safety, as neither functional nor structural abnormalities were noted in diabetic hearts. Ex vivo, hidrosmin induced vascular relaxation that was blocked by nitric oxide synthase (NOS) inhibition. In vitro, hidrosmin stimulated endothelial NOS activity and NO production and downregulated hyperglycemia-induced inflammatory and oxidant genes in vascular smooth muscle cells. Our results highlight hidrosmin as a potential add-on therapy in the treatment of macrovascular complications of diabetes.
RESUMO
Validated biomarkers to be used as biological tools for managing ocular surface diseases (OSDs) are still an unmet need in daily clinical practice. Many studies have contributed to the already extensive list of candidate biomarkers for these disorders. Dry eye (DE) and ocular allergy (OA) are complex and multifactorial diseases, often coexisting and with overlapping symptoms. The purpose of this review is to present a comprehensive updated revision of the most relevant biomarkers of DE and OA, with an emphasis on quantitative analyses and correlations with clinical parameter data. Analysis of biomarkers common for these pathologies has highlighted an important physiological process. Namely, the interleukin proteins (IL-1α, IL-1ß and IL-17), tumour necrotic factor (TNFα) and interferon gamma (IFNγ; Th1-Th7 pathway) and IL-4, IL-5 and IL-13 (Th2 pathway) seem to represent similar inflammatory mechanisms. Moreover, changes in the levels of mucins (MUC1, MUC2, MUC4, MUC5 and MUC16) are common alterations in the tear film mucous layer. We also examine the current state of medical devices and the main limitations to their use in clinical practice. Translational research in biomarkers for clinical practice depends on a feasible transition from the laboratory to the point-of-care. This requires large-scale, coordinated clinical validation campaigns to select the biomarkers with the highest specificity and sensitivity and significant correlation with clinical parameters. Moreover, technical limitations of multiplexed quantitation systems must be overcome to detect and measure the levels of several biomarkers in very small samples. To ensure the future of biomarker research, significant progress is necessary in a number of fields. There is an urgent need for global unification of clinical classification and diagnostics criteria. Widespread integration of proteomic and transcriptomic data is paramount for performing meaningful analyses using appropriate bioinformatics tools and artificial intelligence systems.
Assuntos
Síndromes do Olho Seco , Oftalmopatias , Hipersensibilidade , Biomarcadores/análise , Biomarcadores/metabolismo , Túnica Conjuntiva/metabolismo , Síndromes do Olho Seco/diagnóstico , Síndromes do Olho Seco/metabolismo , Oftalmopatias/diagnóstico , Oftalmopatias/metabolismo , Humanos , Hipersensibilidade/diagnóstico , Hipersensibilidade/metabolismo , Hipersensibilidade/patologia , Interferon gama/metabolismo , Mucinas/metabolismo , Proteômica , Lágrimas/metabolismoRESUMO
Diabetes mellitus (DM) is a high-impact disease commonly characterized by hyperglycemia, inflammation, and oxidative stress. Diabetic nephropathy (DN) is a common diabetic microvascular complication and the leading cause of chronic kidney disease worldwide. This study investigates the protective effects of the synthetic flavonoid hidrosmin (5-O-(beta-hydroxyethyl) diosmin) in experimental DN induced by streptozotocin injection in apolipoprotein E deficient mice. Oral administration of hidrosmin (300 mg/kg/day, n = 11) to diabetic mice for 7 weeks markedly reduced albuminuria (albumin-to-creatinine ratio: 47 ± 11% vs. control) and ameliorated renal pathological damage and expression of kidney injury markers. Kidneys of hidrosmin-treated mice exhibited lower content of macrophages and T cells, reduced expression of cytokines and chemokines, and attenuated inflammatory signaling pathways. Hidrosmin treatment improved the redox balance by reducing prooxidant enzymes and enhancing antioxidant genes, and also decreased senescence markers in diabetic kidneys. In vitro, hidrosmin dose-dependently reduced the expression of inflammatory and oxidative genes in tubuloepithelial cells exposed to either high-glucose or cytokines, with no evidence of cytotoxicity at effective concentrations. In conclusion, the synthetic flavonoid hidrosmin exerts a beneficial effect against DN by reducing inflammation, oxidative stress, and senescence pathways. Hidrosmin could have a potential role as a coadjutant therapy for the chronic complications of DM.
RESUMO
Understanding the regulation of cellular AA uptake as protein supply changes is critical for predicting milk component yields because intracellular supplies partly regulate protein synthesis. Our objective was to evaluate cellular uptake and kinetic behavior of individual AA when cells are presented with varying extracellular AA supplies. Bovine primary mammary epithelial cells were grown to confluency and transferred to medium with an AA profile and concentration similar to that of plasma from dairy cows for 24 h. Treatments were 4 AA concentrations, 0.36, 2.30, 4.28, and 6.24 mM, which represented 16, 100, 186, and 271% of typical plasma AA concentrations, respectively, in lactating dairy cows. Twenty-four plates of cells (89.4 × 19.2 mm) were assigned to each treatment. Cells were first subjected to treatment medium enriched with 15N-labeled AA for 24 h and then incubated with treatment medium enriched with 13C-labeled AA for 0, 15, 60, 300, 900, 1,800, and 3,600 s. Intracellular free AA, intracellular protein-bound AA, and extracellular medium free AA were analyzed for concentrations and isotopic enrichment using gas chromatography-mass spectrometry. A dynamic, 12-pool model was fitted to the data for 14 AA to derive unidirectional uptake and efflux, protein turnover, transamination, oxidation, and synthesis. The derived concentration for half the maximal uptake (km) indicated no saturation of AA uptake at typical in vivo concentrations for 11 of the 14 AA. Arginine, Pro, and Val appeared to exhibit saturation kinetics. Net uptake of all essential AA except Phe was positive across treatments. Most nonessential AA exhibited negative net uptake values. Efflux of AA was quite high, with several AA exhibiting greater than 100% efflux of the respective influx. Intracellular pool turnover was rapid for most AA (e.g., 2 min for Arg), demonstrating plasticity in matching needs for protein translation to supplies. Intracellular AA concentrations increased linearly in response to treatment for most AA, whereas 9 AA exhibited quadratic responses. Amino acid uptake is responsive to varying extracellular supplies to maintain homeostasis. No saturation of uptake was evident for most AA, indicating that transporter capacity is likely not a limitation for most AA except possibly Arg, Val, and Pro in mammary epithelial cells.
Assuntos
Aminoácidos , Lactação , Animais , Bovinos , Células Epiteliais , Feminino , Glândulas Mamárias Animais , Leite , Proteínas do LeiteRESUMO
Understanding uptake of AA by mammary tissue as supply varies is critical for predicting milk component production. Our objective was to develop an in vitro method to quantify cellular uptake, efflux, and intracellular metabolism of individual AA that could be implemented for evaluating these factors when AA supply and profile are varied. Bovine primary mammary epithelial cells were grown to confluency and exposed to medium with an AA profile and concentration similar to lactating dairy cow plasma for 24 h. Cells were then preloaded in medium enriched with 15N-labeled AA for 24 h followed by removal of the 15N-labeled medium and incubation with medium enriched with 13C-labeled AA for 0, 15, 60, 300, 900, 1,800, and 3,600 s. Extracellular free AA and intracellular free and protein-bound AA were analyzed for concentrations and isotopic enrichment by gas chromatography-mass spectrometry. A dynamic, 12-pool model was constructed representing extracellular and intracellular free and protein-bound pools of an AA, and their respective 15N and 13C isotopes. Markov chain Monte Carlo simulation (n = 5,000) was conducted to evaluate prediction errors by deriving standard errors and posterior distributions for rate constants, fluxes, and pools. Cellular Ala influx and efflux were higher than Leu, reflecting Ala role in driving system L transport and the high capacity of sodium-dependent transport. The Ala and Leu turnover rates were 181 and 95, 580 and 857, and 74 and 157% per hour for extracellular, intracellular, and fast protein-bound pools, respectively. The intracellular and extracellular Ala to Leu ratios were quite different, meaning the blood AA profile is not the AA profile provided for protein translation. The high level of exchange and rapid turnover of pools provide a mechanism for matching the AA supplies to the precision necessary for translation. This also understates the importance of using experimental medium similar to what is observed in vivo given that some AA depend on other AA for influx (exchange driven). The average root mean squared prediction error across the isotope enrichments, pools, and concentrations was 9.7 and 14.1% for Ala and Leu, respectively, and collinearity among parameters was low, indicating adequate fit and identifiability. The described model provides insight on individual AA transport kinetics and a method for future evaluation of AA transport and intracellular metabolism when subjected to varying AA supplies.
Assuntos
Aminoácidos/metabolismo , Bovinos , Células Epiteliais/metabolismo , Glândulas Mamárias Animais/citologia , Alanina/metabolismo , Aminoácidos/sangue , Animais , Transporte Biológico , Feminino , Técnicas In Vitro/veterinária , Marcação por Isótopo/veterinária , Cinética , Lactação , Leucina/metabolismo , Glândulas Mamárias Animais/metabolismo , Proteínas do Leite/metabolismoRESUMO
Scalp eschar and neck lymphadenopathy after a tick bite (SENLAT) is an emerging non-pathogen-specific syndrome characterized by scalp eschar and occipital and/or cervical lymph node enlargement following a tick bite. We report a case of SENLAT syndrome in an Argentinian patient after exposure to ticks during outdoor work in the Paraná River Delta region.
Assuntos
Linfadenopatia/etiologia , Dermatoses do Couro Cabeludo/etiologia , Picadas de Carrapatos/complicações , Adulto , Animais , Antibacterianos/uso terapêutico , Argentina , Doxiciclina/uso terapêutico , Humanos , Linfadenopatia/tratamento farmacológico , Masculino , Pescoço , Fotografação , Infecções por Rickettsia/diagnóstico , Dermatoses do Couro Cabeludo/tratamento farmacológico , Dermatoses do Couro Cabeludo/patologia , Picadas de Carrapatos/tratamento farmacológicoRESUMO
Biocrusts are key drivers of the structure and functioning of drylands and are very sensitive to disturbance, including atmospheric nitrogen (N) deposition. We studied the impacts of simulated N deposition on biocrust community composition and soil photosynthetic and photoprotective pigment content after five years of N application in a European semiarid Mediterranean shrubland. The experiment consisted in six experimental blocks with four plots, each receiving 0, 10, 20, or 50 kg NH4NO3-N ha-1 year-1 + 6-7 kg N ha-1 year-1 background. After 5 years of N application, total lichen cover decreased up to 50% compared to control conditions and these changes were only clearly evident when evaluated from a temporal perspective (i.e. as the percentage of change from the first survey in 2008 to the last survey in 2012). In contrast, moss cover did not change in response to N, suggesting that biocrust community alterations operate via species- and functional group-specific effects. Interestingly, between-year variations in biocrust cover tracked variations in autumnal precipitation, showing that these communities are more dynamic than previously thought. Biocrust species alterations in response to N were, however, often secondary when compared to the role of ecologically relevant drivers such as soil pH and shrub cover, which greatly determined the composition and inter-annual dynamics of the biocrust community. Similarly, cyanobacterial abundance and soil pigment concentration were greatly determined by biotic and abiotic interactions, soil pH for pigments, and organic matter content and shrub cover for cyanobacteria. Biocrusts, and particularly the lichen component, are highly sensitive to N deposition and their responses to pollutant N can be best understood when evaluated from a temporal and multivariate perspective, including impacts mediated by interactions with biotic and abiotic drivers.
Assuntos
Briófitas/metabolismo , Líquens/metabolismo , Nitrogênio/química , Solo/química , Cianobactérias/metabolismo , Ecossistema , Nitrogênio/metabolismo , PigmentaçãoRESUMO
Sirtuin 2 (SIRT2) is a member of a family of NAD+ -dependent histone deacetylases (HDAC) that play diverse roles in cellular metabolism and especially for aging process. SIRT2 is located in the nucleus, cytoplasm, and mitochondria, is highly expressed in the central nervous system (CNS), and has been reported to regulate a variety of processes including oxidative stress, genome integrity, and myelination. However, little is known about the role of SIRT2 in the nervous system specifically during aging. Here, we show that middle-aged, 13-month-old mice lacking SIRT2 exhibit locomotor dysfunction due to axonal degeneration, which was not present in young SIRT2 mice. In addition, these Sirt2-/- mice exhibit mitochondrial depletion resulting in energy failure, and redox dyshomeostasis. Our results provide a novel link between SIRT2 and physiological aging impacting the axonal compartment of the central nervous system, while supporting a major role for SIRT2 in orchestrating its metabolic regulation. This underscores the value of SIRT2 as a therapeutic target in the most prevalent neurodegenerative diseases that undergo with axonal degeneration associated with redox and energetic dyshomeostasis.
Assuntos
Axônios/metabolismo , Locomoção/fisiologia , Sirtuína 2/deficiência , Envelhecimento/metabolismo , Envelhecimento/patologia , Animais , Axônios/patologia , Cognição/fisiologia , DNA Mitocondrial/genética , DNA Mitocondrial/metabolismo , Metabolismo Energético , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/metabolismo , Doenças Neurodegenerativas/patologia , Oxirredução , Sirtuína 2/metabolismoRESUMO
PURPOSE: To assess the effects of autologous serum eye drops on conjunctival expression of the mucin gene MUC5AC by means of quantitative reverse transcription polymerase chain reaction (RT-qPCR). METHODS: A prospective and comparative interventional case series study of 38 eyes of 19 patients with different ocular surface disorders was performed before and 6 weeks after the treatment with autologous serum eye drops. All patients underwent a complete ophthalmic assessment, including evaluation of the tear film, ocular surface exploration, conjunctival impression cytology (IC), and MUC5AC detection by reverse transcription polymerase chain reaction. RESULTS: A total of 34 eyes were studied by IC and MUC5AC quantitative reverse transcription polymerase chain reaction before and after treatment with autologous serum. This treatment improved breakup time, conjunctival squamous metaplasia, goblet cell density, and subjective perception in 76.2%, 70.6%, 55.9% and 73.5% of eyes, respectively. Treatment with autologous serum enhanced conjunctival expression of MUC5AC (P = 0.001), although these differences were not statistically significant if data are analyzed patient by patient (P = 0.09). In 13 of 34 eyes (38.2%), we found increased expression of MUC5AC; in 12 eyes (35.3%), no significant changes were found; and in 9 eyes (26.5%), a decreased expression was found. The MUC5AC gene upregulation was related to the conjunctival involvement before treatment and with the improvement in the degree of squamous metaplasia and the increase in the number of goblet cells in IC after treatment (P = 0.001). CONCLUSIONS: Treatment with autologous serum enhances the conjunctival expression of MUC5AC by increasing the density of goblet cells, mainly in patients with severe conjunctival involvement.
Assuntos
Túnica Conjuntiva/metabolismo , Doenças da Túnica Conjuntiva/terapia , Mucina-5AC/metabolismo , Soluções Oftálmicas/uso terapêutico , Soro , Adulto , Idoso , Túnica Conjuntiva/efeitos dos fármacos , Doenças da Túnica Conjuntiva/metabolismo , Doenças da Túnica Conjuntiva/patologia , Feminino , Células Caliciformes/efeitos dos fármacos , Humanos , Masculino , Metaplasia , Pessoa de Meia-Idade , Soluções Oftálmicas/farmacologia , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Lágrimas/efeitos dos fármacos , Lágrimas/metabolismoRESUMO
PURPOSE: To evaluate the sensitivity and specificity of a PCR-strip system based on reverse dot blot for detection of MUC5AC mRNA in corneal epithelium samples from patients with limbal stem cell deficiency (LSCD), and to determine the correlation with clinical diagnosis. METHODS: We obtained 87 corneal impression cytology (IC) samples from 55 subjects (37 patients clinically diagnosed with LSCD and 18 control subjects). Total RNA was extracted from each IC sample and retrotranscribed to cDNA. MUC5AC mRNA transcript was amplified by a customized RT-PCR assay and detected in PCR strips based on reverse dot blot hybridization and in agarose gels. Conjunctival IC samples were used as positive controls. RESULTS: Forty-four of 45 corneal IC samples obtained from patients clinically diagnosed with LSCD were positive for MUC5AC, whereas 34 of 42 corneal ICs from healthy subjects were negative for MUC5AC. Four healthy corneas were found MUC5AC positive, and four rendered inconclusive results. A correlation of 91.4% (P < 0.001) between molecular testing and clinical diagnosis was found. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the PCR-strip system were 98%, 89%, 92%, and 97%, respectively. CONCLUSIONS: Corneal epithelium MUC5AC transcript detection by reverse dot blot PCR-strip is a highly sensitive technique for LSCD diagnosis. The test results strongly correlate with clinical diagnosis of characterized LSCD cases. The PCR-strip system may be used for early detection, and for the detection of mild cases of LSCD, and constitutes an objective clinical tool for the monitoring of treatments and surgical decisions.
Assuntos
Doenças da Córnea/diagnóstico , Epitélio Corneano/patologia , Limbo da Córnea/patologia , Mucina-5AC/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Células-Tronco/patologia , Adulto , Doenças da Córnea/genética , Técnicas de Diagnóstico Oftalmológico , Epitélio Corneano/fisiologia , Feminino , Testes Genéticos/métodos , Testes Genéticos/normas , Humanos , Limbo da Córnea/fisiologia , Masculino , RNA Mensageiro/genética , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Sensibilidade e Especificidade , Células-Tronco/fisiologiaRESUMO
PURPOSE: To investigate the effect of poly (lactic-co-glycolic acid) (PLGA) implants loaded with mitomycin C (MMC) and with different adjuvant treatments after glaucoma filtration surgery (GFS), in comparison to standard treatments. METHODS: Forty-two New Zealand White rabbits underwent bilateral GFS and received different treatments: topical MMC (group 1); topical 5-fluorouracil (5-FU; group 2); PLGA implant (group 3); MMC-loaded and -coated PLGA implant (group 4); MMC-loaded and 5-FU-coated PLGA implant (group 5); subconjunctival bevacizumab (group 6); MMC-loaded PLGA implant and subconjunctival bevacizumab (group 7); and no treatment (right eye of all animals; control group). Intraocular pressure (IOP) and filtering bleb were evaluated on different days after GFS. Histology was performed to examine the conjunctiva, sclerotomy, filtering bleb, and persistence of the implant. RESULTS: The best hypotensive results were achieved in the MMC-loaded and -coated PLGA implant group, which presented the lowest IOP values on days 1, 5, 7, 14, and 28 after GFS. Excluding the implant groups, in which the bleb could not be properly measured, bleb survival was superior to controls in groups 1, 2 and lower in group 6. Group 7 presented greater extension, height, and vascularization of the bleb. Epithelial thinning and lymphoplasmacytic infiltrate were observed in groups 1, 2, 4, 5, and 7. The rates of closure of the sclerotomy and bleb were 100% and 76%, respectively, and implant persistence was 95%. CONCLUSIONS: MMC-loaded and -coated implants have optimal surgical results, followed by topical MMC application. In this experimental model, bevacizumab could interact with MMC.
Assuntos
Antimitóticos/administração & dosagem , Túnica Conjuntiva/cirurgia , Glaucoma/cirurgia , Ácido Láctico , Ácido Poliglicólico , Complicações Pós-Operatórias/tratamento farmacológico , Próteses e Implantes , Trabeculectomia/métodos , Animais , Materiais Revestidos Biocompatíveis , Modelos Animais de Doenças , Portadores de Fármacos , Feminino , Glaucoma/fisiopatologia , Pressão Intraocular , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Coelhos , Resultado do TratamentoRESUMO
Cytogenetic characterization of the neotropical fish Brycon henni (Pisces: Characidae). Brycon henni, is a native fish from Western Colombia is important as food and in sport fishing, and could be cultured in waters between 18 and 28ºC. A previous cytogenetic study in branchial cells indicated different chromosomal complements. Total blood samples were seeded in 4 ml Ham-F12 culture medium, supplemented with 0.5 ml Bovine Fetal Serum and 0.7 ml Phyitohemaglutinin M (Gibco®) during 96 hr at 28ºC; an antimitotic factor (100µl Colcemid® 10%) was added. Cells were incubated in 10 ml KCl hypotonic solution (0.56%) for 24 min at 37ºC, fixed in methanol; acetic acid (3: 1) three times, separated by centrifugations, extended drip, and the chromosomes slides were stained with 5% Giemsa (5%). The best metaphases (6 males and 8 females) were photographed. There was a diploid number of 50 chromosomes: Metacentrics (26M), Submetacentrics (16SM) and Subtelocentrics (8ST). The fundamental number is 100 arms. The evolutionary tendency is type B with no supernumerary chromosomes; a first pair of big metacentric chromosomes is characteristic for Brycon, while no chromosomic sexual heteromorfism was detected. Rev. Biol. Trop. 56 (4): 1619-1628. Epub 2008 December 12.
La sabaleta Brycon henni, es un pez nativo del occidente colombiano importante para la seguridad alimentaria de las poblaciones ribereñas y para la pesca deportiva por sus característica combativas. El desarrollarse entre los 18 y 28ºC la convierte en una especie de cultivo promisoria. Un estudio citogenético a partir de células branquiales indicó diferentes complementos cromosómicos. Este trabajo aplicó la técnica de cultivo de linfocitos a muestras de sangre de adultos. Las muestras de sangre total fueron sembradas en 4 ml de medio de cultivo Ham-F12, suplementado con 0.5 ml de Suero Fetal Bovino y 0.7 ml del mitógeno Fitohemaglutinina M (Gibco®) durante 96 hr a 28ºC; 1.5 hr antes de la cosecha agregamos 100µl de Colcemid al 10% como factor antimitótico. Cada cultivo se incubó con 10 ml de solución hipotónica KCl 0.56% por 24 min a 37ºC, para continuar con tres fijaciones sucesivas, centrifugaciones y tinción Giemsa al 5%. Las mejores metafases fueron fotografiadas en microscopio, correspondientes a 6 machos y 8 hembras, indicando un número diploide de 50 cromosomas, clasificados en Metacéntricos (26M), Submetacéntricos (16SM) y Subtelocéntricos (8ST), para un número fundamental (NF) de 100 brazos. La tendencia evolutiva hallada fue de tipo B; no se encontraron cromosomas supernumerarios pero sí un primer par de cromosomas metacéntricos grandes, considerado marcador para el género Brycon, no determinante de heteromorfismo sexual. Estos resultados coinciden con los demás bricónidos investigados, en donde se podría considerar un ancestro común con un número cromosómico básico y cariotipos simétricos.
Assuntos
Animais , Feminino , Masculino , Análise Citogenética/métodos , Peixes/genética , Colômbia , Cariotipagem/métodosRESUMO
Brycon henni, is a native fish from Western Colombia is important as food and in sport fishing, and could be cultured in waters between 18 and 28 degrees C. A previous cytogenetic study in branchial cells indicated different chromosomal complements. Total blood samples were seeded in 4 ml Ham-F12 culture medium, supplemented with 0.5 ml Bovine Fetal Serum and 0.7 ml Phyitohemaglutinin M (Gibco) during 96 hr at 28 degrees C; an antimitotic factor (100 microl Colcemid 10%) was added. Cells were incubated in 10 ml KCl hypotonic solution (0.56%) for 24 min at 37 degrees C, fixed in methanol; acetic acid (3: 1) three times, separated by centrifugations, extended drip, and the chromosomes slides were stained with 5% Giemsa (5%). The best metaphases (6 males and 8 females) were photographed. There was a diploid number of 50 chromosomes: Metacentrics (26M), Submetacentrics (16SM) and Subtelocentrics (8ST). The fundamental number is 100 arms. The evolutionary tendency is type B with no supernumerary chromosomes; a first pair of big metacentric chromosomes is characteristic for Brycon, while no chromosomic sexual heteromorfism was detected.