27 research outputs found

    Synthesis, characterization and application of ion imprinted poly(vinylimidazole) for zinc ion extraction/preconcentration with faas determination

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    In this paper, we describe the synthesis of an ion imprinted polymer (IIP) by homogeneous polymerization and its use in solid-phase to extract and preconcentrate zinc ions. Under optimal conditions (pH 5.0, preconcentration flow rate of 12.0 mL min-1, and eluted with 1.0 mol L-1 HNO3) this procedure allows the determination of zinc with an enrichment factor of 10.2, and with limits of detection and quantification of 1.5 and 5.0 µg L-1, respectively. The accuracy of our results was confirmed by analysis of tap water and certified reference materials: NIST 1570a (Spinach leaves) and NIST 1515 (Apple leaves)371636

    A New and Fast Technique to Generate Offspring after Germ Cells Transplantation in Adult Fish: The Nile Tilapia (Oreochromis niloticus) Model

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    Background: Germ cell transplantation results in fertile recipients and is the only available approach to functionally investigate the spermatogonial stem cell biology in mammals and probably in other vertebrates. In the current study, we describe a novel non-surgical methodology for efficient spermatogonial transplantation into the testes of adult tilapia (O. niloticus), in which endogenous spermatogenesis had been depleted with the cytostatic drug busulfan. Methodology/Principal Findings: Using two different tilapia strains, the production of fertile spermatozoa with donor characteristics was demonstrated in adult recipient, which also sired progeny with the donor genotype. Also, after cryopreservation tilapia spermatogonial cells were able to differentiate to spermatozoa in the testes of recipient fishes. These findings indicate that injecting germ cells directly into adult testis facilitates and enable fast generation of donor spermatogenesis and offspring compared to previously described methods. Conclusion: Therefore, a new suitable methodology for biotechnological investigations in aquaculture was established, with a high potential to improve the production of commercially valuable fish, generate transgenic animals and preserv

    Preparation of a new restricted access molecularly imprinted hybrid adsorbent for the extraction of folic acid from milk powder samples

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    Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)A new hybrid molecularly imprinted polymer (MIP) combined with restricted access (RAM) has been synthesized based on random free-radical polymerization and a sol-gel process. From competitive adsorption studies by using the RAM-MIP and RAM-NIP with folic acid and structurally similar molecules (caffeine, 4-aminobenzamide or paracetamol), relative selectivity coefficients (k) higher than one unit were achieved, indicating good recognition selectivity for folic acid. The percentage of BSA protein exclusion was higher for the RAM-MIP (55.3 + 2.0) as compared to the MIP (35.9 + 2.5%). The solid phase extraction (SPE) procedure was performed by loading the acid extract from milk powder samples previously submitted to saponification and acidification with 10% trichloroacetic acid (TCA) until pH 1.5, through 100 mg of RAM-MIP packed into SPE cartridge. The elution step was carried out by using a mixture of acetonitrile : 0.266 mol L-1 acetate buffer at pH 5.7 (15 : 85, v/v), the same composition of the mobile phase of HPLC. The intra-day precision (n = 10) of the procedure assessed as relative standard deviation (RSD) was 4.7 and 4.1% for the respective concentrations of 20.0 and 150.0 mu g L-1. The applicability of the method was attested by analysis of different brands of milk powder samples fortified with folic acid, as well as by high recovery percentages 95.0-108.4% obtained upon addition and recovery tests. The cleanup process accomplished by RAM-MIP was so efficient that very few remaining matrix components were detected in the eluate by high performance liquid chromatography.A new hybrid molecularly imprinted polymer (MIP) combined with restricted access (RAM) has been synthesized based on random free-radical polymerization and a sol–gel process. From competitive adsorption studies by using the RAM-MIP and RAM-NIP with folic83656665COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO ARAUCÁRIA DE APOIO AO DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO DO PARANÁ - FACONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)sem informaçãosem informaçãosem informaçãoWe acknowledge the CAPES, INCT-BIOANALITICA, FUNDAÇÃO ARAUCÁRIA do PARANÁ, SETI do Paraná and CNPq for financial support. The authors also would like to thank all the anonymous reviewers for their valuable comments and suggestions to improve the quality

    Duration of spermatogenesis in the bullfrog (Lithobates catesbeianus)

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    The bullfrog (Lithobates catesbeianus) has substantial economic importance and has also been used as an experimental model for biological studies in the fields of pharmcology, medicine, and reproductive biology, especially studies addressing gametogenesis. However, there is a lack of comprehensive information in the literature regarding testis structure and function in this amphibian. The main objective of the current study was to estimate the duration of the various phases of spermatogenesis in this vertebrate. Sixteen sexually mature bullfrogs received an intracoelomic administration of tritiated thymidine. Testes were analyzed at various times between I h and 33 d after administration to detect the most advanced germ cell types labeled at each interval, as well as labeled preleptotene spermatocytes, which presumably originated from spermatogonial stem cells. The duration of the spermatogonial, spermatocytic, and spermiogenic phases of spermatogenesis in the bullfrog were approximately 18, 14, and 8 d, respectively. Thus, the total duration of the spermatogenesis process from early spermatogonia through to spermatozoa was 40 d in this species, similar to that of most previously investigated mammalian species. To our knowledge, this is the first reliable report on the duration of the full spermatogenic process in any amphibian species. These findings will be very useful for tracking the pace of germ cells in studies involving spermatogonial transplantation in lower vertebrates. (C) 2009 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

    Ultrastructure of the urethra of the Mongolian gerbil

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    Objective: the urethra is the main port of entry of sexually transmitted pathogens. However, papers on the morphology of the urethra are scarce. The Mongolian gerbil is a rodent native of the Mongolia and China and has been utilized as a laboratory animal since the 1960s. This work describes the ultrastructure of the urethra of the Mongolian gerbil to provide data for future experimental studies. Methods: the urethra of ten adult male gerbils was studied by transmission electron microscopy. Results: the epithelium of the pelvic urethra possesses two cell types: I and II, without the formation of cellular layers, while the penile urethra possesses cellular layers: basal, intermediate and superficial. The urethra presents neurosecretory cells belonging to the amine precursor uptake and decarboxylation system. Conclusions: the urethral epithelium of the gerbil is a neurosecretory epithelium, part of the amine precursor uptake and decarboxylation system

    Ultrastructural study of acrosome formation in mongolian gerbil (Meriones unguiculatus)

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    Spermatogenesis is a complex and very well organized process lasting from 30 to 75 days in mammals. The spermatogenic process has been described mainly in laboratory mammals, such as the rat, while correspondent studies in wild animals are scarce. The gerbil (Meriones unguiculatus) is a small rodent native of the arid regions of Mongolia and China. Few reports are available on reproduction in the male Mongolian gerbil. The present study provides the first description of the ultrastructural alterations in spermatid cytoplasm and nucleus, with particular reference to acrosome formation in gerbils. The testes were processed by conventional transmission electron microscopy technique. Based on the development of the acrosomal system and changes in nuclear morphology, the transformation of spermatids in spermatozoon was divided into 15 steps. There were four phases in the spermiogenesis process in the gerbil: Golgi, cap, acrosomal and maturation phases. This provides the foundation for a variety of future studies of the spermiogenesis of this animal. (C) 2000 Harcourt Publishers Ltd

    Spermatogenic cycle length and spermatogenic efficiency in the gerbil (Meriones unguiculatus)

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    The gerbil (Meriones unguiculatus) is a rodent native of the and regions of Mongolia and China. Because the gerbil can be easily bred in laboratory conditions, this species has been largely used as an experimental model in biomedical research. However, there is still little information concerning the testis structure and function in the gerbil. In this regard, we performed a detailed morphofunctional analysis of the gerbil testis and estimated the spermatogenic cycle length utilizing H-3-thymidine as a marker for germ cell progression during their evolution through the spermatogenic process. The stage frequencies of the XII stages characterized according to the acrosome formation and development were (I-XII) 13.8, 10.1, 8.1, 7.8, 4.0, 11.2, 7.5, 7.1, 5.9, 7.6, 8.1, and 8.9. The mean duration of each seminiferous epithelium cycle was determined to be 10.6 +/- 1.0 days and the total duration of spermatogenesis, based on 4.5 cycles, was approximately 47.5 days. The volume density of tubular and interstitial compartments was approximately 92% and 8%, respectively. Based on the volume occupied by seminiferous tubules in the testis and the tubular diameter, about 9 and 18 m of seminiferous tubules were found per testis and per gram of testis, respectively. Twelve primary spermatocytes were formed from each type A1 spermatogonia. The meiotic index was 2.8, indicating that 30% of cell loss occurs during meiosis. The number of Leydig and Sertoli cells per gram of the testis was 28 million and each Sertoli cell was able to support approximately 13 spermatids. The daily sperm production per gram of testis (spermatogenic efficiency) was 33 million. Taken together, these data indicate that, mainly due to the high seminiferous tubule volume density and Sertoli cell support capacity for germ cells, the gerbil presents high spermatogenic efficiency compared with other mammalian species already investigated. The data obtained in the present study might provide the basis for future research involving the reproductive biology in this species
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