RESUMEN
Echinococcosis caused by the larval form of Echinococcus granulosus (E. granulosus) is known as an important zoonotic disease in various parts of the world, including Iran. The genetic diversity of this parasite is very high, particularly in areas where the disease is endemic. It has been suggested in the literature from different parts of the world that diverse factors, such as parasite life cycle, transmission pathways, pathologic disease, immunization, and disease control can be affected by the genetic diversity of the parasite. Various studies indicated sheep strain G1 as the most common genotype throughout the world. This strain is commonly found in the liver and lung repeatedly causing echinococcosis in humans, sheep, and cattle. The present study was conducted to determine the genetic affinity between the protoscolex of E. granulosus in humans and sheep in East Azerbaijan province, Iran for the first time. A total of 120 hydatid cyst samples were collected, 60 of which were from people who referred to the hospitals of East Azerbaijan and 60 were from the sheep slaughtered in Tabriz slaughterhouse. Following DNA extraction, certain regions of the cox1 gene were amplified and evaluated by the polymerase chain reaction. The replicated parts in all isolates had the same size of 450 bp. Electrophoresis was followed by selecting a total of 60 suitable samples, including 30 human samples and 30 sheep samples and sending them for genome sequencing. The overlap of the samples was investigated using the BLAST software. The results of BLAST, sequencing, and overlap demonstrated a genetic linkage of approximately 91.76% between the protoscolex of E. granulosus in human and sheep.
Asunto(s)
Equinococosis/parasitología , Equinococosis/veterinaria , Echinococcus granulosus/genética , Enfermedades de las Ovejas/parasitología , Animales , Echinococcus granulosus/crecimiento & desarrollo , Humanos , Irán , Larva/genética , OvinosRESUMEN
Embryo transfer is a reproductive technique that has a major impact on the dissemination of economically important genes and the rate of genetic gain in breeding schemes. In recent years, there has been increasing interest in the use of sexed and genotyped embryos in commercial embryo transfer programs. Marker/gene assisted selection (MAS/GAS) projects can be performed in the pre-implantation stage through mass production of characterized embryos. Biopsy of a few cells in the morulla stage is essential for pre-implantation genetic diagnosis (PGD), in which sex determination, evaluation of disease genes, and genotyping for candidate genes are performed. Limited quantity of cells and low amount of DNA restrict the use of multiple molecular analyses in PGD programs. Recently, whole genome amplification (WGA) techniques promise to overcome this problem by providing sufficient input DNA for analysis. Among several techniques proposed for WGA, the primer extension pre-amplification (PEP) and the improved-primer extension pre-amplification (I-PEP) methods are the most commonly used. However, these methods are time-consuming and need more than 12 h amplification cycles. Since the time is a critical parameter in the successful characterized embryo transfer, the shortening of diagnosis time is highly desirable. In this study, we developed a short and simple I-PEP procedure (~3 h) and evaluated its performance for the amplification of bovine genomic DNA. We assessed short WGA procedure by polymerase chain reaction (PCR) amplification of 7 specific loci. The results indicated that the short procedure possesses enough sensitivity for the molecular genetic analysis of 1 input cell. Although the efficiency of the method was 100%, there was an inconsistency between genomic DNA (gDNA) and whole genome amplification product (wgaDNA) genotypes for kappa-casein locus; that is, however, most likely due to allele drop-out (ADO) or false homozigocity. The results of this study indicate that with the application of reliable methods, WGA-amplified bovine DNA will be a useful source for sexing and genotyping bovine embryos in several quantitative trait locus (QTL) markers.
Asunto(s)
Blastocisto/fisiología , Bovinos/genética , Genoma , Genómica/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Animales , Bovinos/embriología , ADN/química , Cartilla de ADN , Electroforesis en Gel de Agar , Transferencia de Embrión , Femenino , Marcadores Genéticos/genética , Masculino , Reacción en Cadena de la PolimerasaRESUMEN
INTRODUCTION: Acute diarrhea disease is the second cause of death among all infectious diseases in children younger than 5 years of age worldwide. The aim of this study was to employ a combination of biochemical, microbiological and molecular diagnostic techniques to investigate the stools of Iranian children with acute diarrhea for bacterial enteropathogens. METHOD: Diarrheagenic Escherichia coli, Shigella spp., Salmonella spp., Campylobacter spp. and Yersinia spp., were investigated from June 2003 to June 2005, in 1087 children less than 5 years old with acute diarrhea. Stool specimens from children were studied for enteropathogens both by standard culturing and molecular methods. This study was designed on hospital based. RESULT: The highest incidence values were found in the summer and in children less than 1-year-old (42.7%). The Pathogenic bacteria recovered out from fecal samples of 555 (55.1%) patients had the following profile: Shigella spp. (26.7%) was the most prevalent bacterial pathogen and Shiga-like toxin producing E. coli (STEC) and Enteroaggregative E. coli (EAEC) 105 (18.9%) and 92 (16.6%) had the second and third highest prevalence, respectively. Enteropathogenic E. coli (EPEC), Campylobacter, Salmonella and Enterotoxigenic E. coli (ETEC) were found in 70 (12.6%), 60 (10.8%), 42 (7.6%), and 38 (6.8%) positive samples, respectively. In this study neither Yersinia nor E. coli O157:H7 were found. Of the 30 co-infections detected, Shigella flexneri and Campylobacter jejuni accounted for more than 50%. CONCLUSION: Information about the prevalence of wide-range Shigella and STEC may facilitate the control and management of infant diarrhea diseases in Iran. The results of this study suggest that comprehensive surveys are needed in different parts of the country in order to identify the incidence of different enteropathogenic diarrhea, especially diarrheagenic E. coli in children in Iran.
Asunto(s)
Infecciones por Campylobacter/epidemiología , Diarrea/epidemiología , Diarrea/microbiología , Infecciones por Enterobacteriaceae/epidemiología , Infecciones por Campylobacter/microbiología , Preescolar , Enterobacteriaceae/aislamiento & purificación , Infecciones por Enterobacteriaceae/microbiología , Heces/microbiología , Hospitales , Humanos , Incidencia , Lactante , Recién Nacido , Irán/epidemiología , Prevalencia , Estaciones del AñoRESUMEN
National registries provide valuable bases for comparisons and aid innovative policies. Iran, with 70 million inhabitants, has a governmentally supported system of organ and tissue procurement and transplantation. By two acts of legislation, first in 1997 regarding governmental rewarding living kidney donation and second in 2000 regarding cadaveric transplantation, this field has had remarkable growth in the last decade, under the supervision of the Ministry of Health. Since the beginning of each organ or tissue transplantation activity up to 2006, 21,359 kidneys, 338 livers, 122 hearts, 20 lungs, 7 pancreas, 1898 bone marrows, 28,838 corneas, 1380 heart valves, and 1366 bones were transplanted in our country. Increases in the number of live kidney transplantations are taking place, but cannot keep pace with the increasing number of patients. It is highly recommended that we try to increase cadaveric transplantation in coming years.
Asunto(s)
Trasplante de Riñón/estadística & datos numéricos , Trasplante de Órganos/estadística & datos numéricos , Cadáver , Trasplante de Corazón/estadística & datos numéricos , Humanos , Irán , Trasplante de Hígado/estadística & datos numéricos , Donadores Vivos/estadística & datos numéricos , Trasplante de Pulmón/estadística & datos numéricos , Sistema de Registros , Donantes de Tejidos/estadística & datos numéricosRESUMEN
The impact of the neutron activation procedure, i.e. incorporation of samarium oxide (Sm(2)O(3)) and neutron irradiation, on the compression properties (including the crushing strength) and in vitro dissolution of potential colonic delivery systems based on matrix tablets of amidated pectin (Am.P) or two types of hydroxypropyl methylcellulose (HPMC) was investigated. The neutron activation factors did not influence the compression properties of the tablets. Replacement of magnesium stearate with samarium stearate in directly compressed Am.P tablets to achieve both radiolabelling and lubrication resulted in a greater extent of concentration-dependent reduction of the crushing strength. Dissolution tests demonstrated that irradiation increased the release of the model drug ropivacaine from the tablets. The extent of this increase was unexpectedly low considering the previously observed degradation of the polymer expressed as an irradiation-induced viscosity reduction in solutions prepared from the polymers. Delayed-release coating with Eudragit L 100 protected the HPMC tablets against the release-increasing effect of irradiation until the late phases of release. Sm(2)O(3) retarded the release to a varying extent depending on particle characteristics. Incorporation of Sm(2)O(3) in the coating layer did not influence the release. However, one-third of the radioactivity leached from the coating within 60 min in 0.1 M HCl.
Asunto(s)
Colon/metabolismo , Neutrones , Comprimidos/efectos de la radiación , Amidas/administración & dosificación , Amidas/farmacocinética , Análisis de Varianza , Fenómenos Químicos , Química Física , Composición de Medicamentos , Excipientes , Dureza/efectos de la radiación , Lactosa/análogos & derivados , Metilcelulosa/análogos & derivados , Microscopía Electrónica de Rastreo , Oxazinas , Pectinas , Ácidos Polimetacrílicos , Ropivacaína , Samario/administración & dosificación , Samario/farmacocinética , Solubilidad , Estearatos/administración & dosificación , Estearatos/farmacocinéticaRESUMEN
The objective of this work was to develop pectin-based matrix tablets for colonic delivery of the model drug ropivacaine, with the future perspective of radiolabelling the system by neutron activation technique for a gamma-scintigraphic study. The aim was to investigate some formulation factors that could reduce the release of the drug in the simulated gastric and intestinal fluids, increase the release in the simulated cecal fluid (with pectinolytic enzymes) and improve the poor compactibility of pectins. For dissolution studies, the flow-through apparatus with sequential dissolution liquids simulating the mouth-to-colon conditions was used. The effect of two pectin types, the incorporation of ethylcellulose as a dry matrix-additive and water or ethanol as granulation liquids were investigated in a study designed as a D-optimal mixture. Amidated pectin (Am.P) produced harder tablets than the calcium salt of pectin (Ca.P) and was more susceptible to enzymatic degradation. Addition of ethylcellulose increased the tablet strength and the dissolution rate. Furthermore, directly compressed Am.P tablets were produced by addition of coarse or micronised qualities of ethylcellulose. The latter improved the crushing strength markedly imposing a marginal release-reducing effect. Coating this formulation with Eudragit((R)) L 100 reduced the release in the simulated upper GI conditions without interference with the subsequent enzymatic activity.
Asunto(s)
Amidas/farmacocinética , Anestésicos Locales/farmacocinética , Sistemas de Liberación de Medicamentos , Pectinas/farmacocinética , Amidas/administración & dosificación , Anestésicos Locales/administración & dosificación , Química Farmacéutica , Colon , Tamaño de la Partícula , Pectinas/química , Ropivacaína , Comprimidos RecubiertosRESUMEN
The effect of the neutron activation factors, i.e., admixture of samarium oxide (Sm2O3) and irradiation time, on the physico-chemical properties of the raw materials and the in vitro dissolution and disintegration of hydrophilic and lipophilic suppositories was investigated. It was possible to expose the pure bases and the model drugs (5-aminosalicylic acid [5-ASA] and ropivacaine hydrochloride) to 1 min of neutron irradiation in a flux of 1.1.1013 n cm-2s-1. The dissolution and disintegration of the corresponding suppositories showed that the physico-chemical properties and the fraction of incorporated drug together with the lipophilic/hydrophilic nature of the base were important factors. Sm2O3 increased the disintegration time of hydrophilic suppositories containing 5-ASA, while the dissolution of both drugs from these formulations remained unchanged. Sm2O3 did not alter the disintegration time of the lipophilic formulations, but it reduced the dissolution of both drugs from these suppositories. Irradiation induced different behaviour in the different bases.
Asunto(s)
Excipientes/efectos de la radiación , Neutrones , Óxidos , Samario , Supositorios/efectos de la radiación , Amidas/química , Amidas/efectos de la radiación , Excipientes/química , Reología , Ropivacaína , Ácido Salicílico/química , Ácido Salicílico/efectos de la radiación , Solubilidad , Supositorios/química , Factores de TiempoRESUMEN
Different excipients, which are currently being studied for colon delivery systems, were examined with respect to their stability toward neutron irradiation as a potential method of radiolabeling the formulations for gamma-scintigraphic studies. Three different pectin and four different hydroxypropyl methylcellulose (HPMC) types, in addition to two types of polymethacrylate films, were exposed to 1, 2, and 3 min of thermal neutron irradiation in a flux of 1.1 x 10(13) n cm-2 s-1. The physicochemical characteristics of pectins and HPMCs and the mechanical properties of the polymethacrylate films were examined after the radioactivity of the samples had declined to background levels. Methods included ultraviolet (UV) and Fourier transform infrared (FTIR) spectroscopy, pH measurements, loss on drying, thermogravimetric analysis (TGA), viscosimetry, gas chromatographic (GC) analysis of pectin monosaccharides, and tensile strength testing of the films. The results suggest that pectins and HPMCs undergo degradation, as expressed by a significant reduction in the dynamic and intrinsic viscosities of the samples. Generally, HPMCs were more sensitive than pectins to neutron irradiation. However, calcium pectinate proved to be the most sensitive among all the investigated polymers. Both polymethacrylate films (Eudragit L and S) resisted loss of mechanical properties following 1 and 2 min of neutron irradiation, whereas irradiation for 3 min implied significant changes in the appearance and the mechanical properties of Eudragit L films. As a conclusion, neutron irradiation results in dose-dependent degradation of the investigated polysaccharides and polymethacrylates. The consequences on the in vitro behavior of a formulation containing such polymers are discussed.
Asunto(s)
Excipientes/efectos de la radiación , Lactosa/análogos & derivados , Metilcelulosa/análogos & derivados , Neutrones , Pectinas/efectos de la radiación , Ácidos Polimetacrílicos/efectos de la radiación , Cromatografía de Gases , Estabilidad de Medicamentos , Concentración de Iones de Hidrógeno , Lactosa/efectos de la radiación , Metilcelulosa/efectos de la radiación , Oxazinas , Pectinas/análisis , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Resistencia a la Tracción , TermogravimetríaRESUMEN
A on the polymerase chain reaction (PCR) method based was developed for detection of Listeria monocytogenes in milk samples after enrichment culture. It consists of culturing samples in Listeria enrichment broth, followed by DNA extraction and detection of the organism using PCR. Dilutions of L. monocytogenes in milk were subjected to PCR amplification after enrichment culture. When determining the sensitivity of the method, it was found to be possible to detect 37 CFU (colony forming unit gl/ml) of the bacterium in milk. The method was assessed as a sensitive, specific, times-saving and practical way of detecting L. monocytogenes in milk samples.
Asunto(s)
Listeria monocytogenes/aislamiento & purificación , Leche/microbiología , Reacción en Cadena de la Polimerasa/métodos , Animales , Bovinos , ADN Bacteriano/análisis , Listeria monocytogenes/genética , Sensibilidad y EspecificidadRESUMEN
One hundred raw milk samples from different regions of Anatolia and 20 pasteurized milk samples from three different manufacturers in Ankara were analyzed for the presence of Listeria spp. L. monocytogenes was found in 1% of the raw milk samples and in 5% (1/20) of the pasteurized milk samples. L. innocua and L. seeligeri were found in 8 and 2% of the raw milk samples, respectively. No other species of Listeria was found. The overall incidence of Listeria spp. was 10% in the raw milk samples and 5% in the pasteurized milk samples.