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Biomedical subjects

A P Fernandes

Publications and source records attributed to A P Fernandes.

At least 19 recordsLinked to original sources
PubMed2005

Phenotypic heterogeneity in biochemical parameters correlates with mutations in AGPAT2 or Seipin genes among Berardinelli-Seip congenital lipodystrophy patients.

The Berardinelli-Seip congenital lipodystrophy (BSCL) syndrome is characterized by a near-total congenital absence of fat and predisposition to develop diabetes mellitus. We have previously reported that 22 patients from 16 consanguineous pedigrees living in the northeastern region of Brazil had a homozygous 669insA mutation in the Seipin gene (BSCL2 locus), while all of the 10 investigated subjects from the southeastern region were homozygous for a 1036 bp deletion in the AGPAT2 gene (BSCL1 locus). In this study, we compared the serum insulin and insulin resistance (HOMA), leptin, triglyceride and fasting glucose levels in individuals of these two genetically distinct clusters of BSCL subjects. The onset of diabetes was also estimated. The fasting glucose and triglyceride levels were not significantly different in these groups. Significant differences were detected for leptin, insulin and insulin resistance. BSCL1 patients presented lower serum leptin levels compared to BSCL2 patients. BSCL2 subjects had earlier onset of diabetes and higher insulin levels. In agreement, BSCL2 patients were more insulin resistant, as detected by HOMA. These results indicate phenotypic heterogeneity between BSCL1 and BSCL2 Brazilian subjects.

Adolescent↗
PubMed2005

Immunocytochemical localization of tubulins in spermatids and spermatozoa of Euptoieta hegesia (Lepidoptera: Nymphalidae).

A comparative analysis of the distribution of tubulin types in apyrene and eupyrene sperm of Euptoieta hegesia butterflies was carried out, also verifying the presence of tubulin in lacinate appendages of the eupyrene sperm. Ultrathin sections of LR White embedded spermatids and spermatozoa were labeled for alpha, beta, gamma, alpha-acetylated and alpha-tyrosinated tubulins. Apyrene and eupyrene spermatids show the same antibody recognition pattern for tubulins. All tubulin types were detected in axonemal microtubules. Alpha and gamma tubulins were also detected on the cytoplasmic microtubules. However, for beta and tyrosinated tubulins only scattered labeling was detected on cytoplasmic microtubules and acetylated tubulin was not detected. In apyrene and eupyrene spermatozoa only the axoneme labeling was analyzed since cytoplasmic microtubules no longer exist in these cells. Alpha, beta and tyrosinated tubulins were easily detected on the apyrene and eupyrene axoneme; gamma tubulin was strongly marked on eupyrene axonemes but was scattered on the apyrene ones. Acetylated tubulin appeared with scattered labeling on the axoneme of both sperm types. Our results demonstrate significant differences in tubulin distribution in apyrene and eupyrene axonemal and cytoplasmic microtubules. Extracellular structures, especially the lacinate appendages, were not labeled by antibodies for any tubulin.

Animals↗
PubMed2003

Wortmannin enhances the induction of micronuclei by low and high LET radiation.

In mammalian cells, the repair of DNA double-strand breaks (DSBs) is mainly mediated by DNA non-homologous end joining. DNA-dependent protein kinase (DNA-PK), a nuclear serine-threonine kinase and a member of the phosphaditylinositol-3 kinase-related kinase family that is activated by DSBs, is a key component of this pathway. Wortmannin (WM) is known to be an irreversible and potent inhibitor of DNA-PK and has thus been proposed as an effective sensitizer for ionizing radiation and for radiomimetic compounds. The present study, using the cytokinesis block micronucleus assay, reports on the differential effect of WM on the repair of the DNA damage induced by low LET ((60)Co gamma-radiation) and high LET radiation by the boron neutron capture reaction (alpha and Li particles) in V79 Chinese hamster cells. Significant increases in the number of micronuclei per binucleated cell as well as in the frequency of micronucleated binucleated cells were observed in the presence of different concentrations of WM for high LET radiation from the boron neutron capture reaction. The increases observed reached a maximum of approximately 2-fold in comparison with the respective controls. WM, however, had a more pronounced effect on (60)Co gamma-radiation-induced micronuclei, increasing the genotoxic damage from this radiation by approximately 3- to 4-fold. These results are in general in agreement with the concept that DSBs induced by high LET radiation are not a more suitable substrate for the end joining processes mediated by DNA-PK, yet they do not preclude a role for DNA-PK in high LET-induced damage repair.

Alpha Particles↗
PubMed2001

Combined interleukin-12 and topical chemotherapy for established Leishmaniasis drastically reduces tissue parasitism and relapses in susceptible mice.

The efficacy of the association of paromomycin sulfate (PA) with recombinant (r) interleukin (IL)-12 was investigated by topical treatment of BALB/c mice infected with Leishmania major that displayed fully developed cutaneous lesions. Although healing was observed in PA-treated groups, lesions recurred in 100% of these animals 70 days after treatment. In contrast, lesions were absent in a high proportion of PA- and rIL-12-treated mice 120 days after treatment. The PA/rIL-12-treated mice had a switch in cytokine response, from high IL-4 and low interferon (IFN)-gamma levels to low IL-4 and high IFN-gamma levels, and reductions in parasite load, dissemination of parasites, and inflammation. Thus, the association of rIL-12 to topical chemotherapy for leishmaniasis may be an important strategy for increasing cure rates and decreasing the incidence of relapse.

Administration, Cutaneous↗
PubMed2001

Brazilian population database for the 13 STR loci of the AmpFlSTR Profiler Plus and Cofiler multiplex kits.

Allele frequencies for the 13 STR core loci (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, CSF1PO, TPOX, THO1 and D16S539) included in the AmpFlSTR((R)) Profiler Plus and AmpFlSTR((R)) Cofiler kits were obtained for a sample of 700-800 genetically unrelated Brazilians. The expected performance of these loci for personal identification and paternity testing in the Brazilian population was estimated.

Alleles↗
PubMed2001

Nuclear changes and acrosome formation during spermiogenesis in Euchistus heros (Hemiptera: Pentatomidae).

Ultrastructural and cytochemical studies were carried out on nuclear changes and acrosome formation during the spermiogenesis of the phytophagous bug Euchistus heros. The development of the nucleus involves changes in the shape and in degree of chromatin condensation: initially it is dispersed and with a low-electron density, then assumes a fibrillar arrangement and finally compacts in an electron-dense material. The acrosome is formed by the Golgi complex and presents unusual morphological features during its development. The reaction product of acid phosphatase, glucose-6-phosphatase and thiamine pyrophosphatase activities were detected during various stages of acrosome development. In contrast, residues of alpha-N-acetylgalactosamine and basic proteins were only reported in the intermediate and late stages of the differentiation process, respectively.

Acid Phosphatase↗
PubMed2001

Evaluation of the genotoxic effects of the boron neutron capture reaction in human melanoma cells using the cytokinesis block micronucleus assay.

The present work reports on the genotoxicity of the boron neutron capture (BNC) reaction in human metastatic melanoma cells (A2058) assessed by the cytokinesis block micronucleus assay (CBMN) using p-borono-L-phenylalanine (BPA) as the boron delivery agent. Different concentrations of BPA (0.48, 1.2 and 2.4 mM) and different fluences of thermal neutrons were studied. Substantial genotoxic potential of alpha and lithium particles generated inside or near the malignant cell by the BNC reaction was observed in a dose-response manner as measured by the frequency of micronucleated binucleated melanoma cells and by the number of micronuclei (MN) per binucleated cell. The distribution of the number of MN per micronucleated binucleated cell was also studied. The BNC reaction clearly modifies this distribution, increasing the frequency of micronucleated cells with 2 and, especially, > or =3 MN and conversely decreasing the frequency of micronucleated cells with 1 MN. A decrease in cell proliferation was also observed which correlated with MN formation. A discrete genotoxic and anti-proliferative contribution from both thermal neutron irradiation and BPA was observed and should be considered secondary. Additionally, V79 Chinese hamster cells (chromosomal aberrations assay) and human lymphocytes (CBMN assay) incubated with different concentrations of BPA alone did not show any evidence of genotoxicity. The presented results reinforce the usefulness of the CBMN assay as an alternative method for assessment of the deleterious effects induced by high LET radiation produced by the BNC reaction in human melanoma cells.

Animals↗
PubMed2001

Spermiogenesis in Melanophryniscus cambaraensis (Amphibia, Anura, Bufonidae): ultrastructural and cytochemical studies of carbohydrates using lectins.

Spermiogenesis in the bufonid Melanophryniscus cambaraensis was analysed ultrastructurally. The nucleus of spermatids undergoes morphological modifications divided into five distinct stages based on chromatin condensation. The acrosomal vesicle differentiates into an acrosome and a subacrosomal cone, which cap the anterior region of the nucleus. The spermatozoa consist of the head with a nucleus and acrosomal complex and the tail, with an axoneme, an axial rod and an undulating membrane. The participation of several carbohydrate moieties during this differentiation process was detected using various gold-labelled lectins.

Animals↗
PubMed2001

Detection of calcium and calmodulin during spermiogenesis of phytophagous bugs (Hemiptera: Pentatomidae).

The ultrastructural detection of calcium using pyroantimonate, and the immunocytochemical localization of calmodulin using monoclonal antibody were carried out during the spermiogenesis of phytophagous bugs. The presence of calcium was observed on the Golgi apparatus during the initial phases of spermiogenesis. In the other stages the calcium was observed in association with the nucleus and in some regions of acrosome. Indeed, it was detected surrounding the mitochondrial derivatives and specific axonemal microtubules on the tail region. The immunocytochemical detection of calmodulin showed the presence of this protein approximately in the same regions where the calcium was detected, indicating that calcium and calmodulin could work together during spermiogenesis of this phytophagous bugs, suggesting their involvement on the regulation of flagellar beating, nuclear compactation and acrosome formation.

Animals↗
PubMed2000

Glucose-mediated Ca(2+) signalling in single clonal insulin-secreting cells: evidence for a mixed model of cellular activation.

Using clonal insulin-secreting BRIN-BD11 cells, we have assessed whether the graded response of the whole cell population to glucose can be accounted for by a dose-dependent recruitment of individual cells, an amplification of the response of the recruited cells or both. Cytosolic free Ca(2+) concentration ([Ca(2+)](i)) is an established index of beta-cell function. We used fura-2 microfluorescence techniques to assess the [Ca(2+)](i) responsiveness of single BRIN-BD11 cells to glucose and other secretagogues. Glucose (1-16.7 mM) evoked oscillatory [Ca(2+)](i) rises in these cells resembling those found in parental rat pancreatic beta-cells. The percentage of glucose-responsive cells was 11% at 1 mM and increased to 40-70% at 3-16.7 mM glucose, as assessed by a single-stimulation protocol. This profile was unrelated to possible differences in the cell cycle, as inferred from experiments where the cultured cells were synchronized by a double thymidine block protocol. Individual cells exhibited variable sensitivities to glucose (threshold range: 1-5 mM) and a variable dose-dependent amplification of the [Ca(2+)](i) responses (EC(50) range: 2-10 mM), as assessed by a multiple-stimulation protocol. Glyceraldehyde and alpha-ketoisocaproic acid had glucose-like effects on [Ca(2+)](i). The data support a mixed model for the activation of insulin-secreting cells. Specifically, the graded secretory response of the whole cell population is likely to reflect both a recruitment of individual cells with different sensitivities to glucose and a dose-dependent amplification of the response of the recruited cells.

Animals↗
PubMed2000

Ultrastructural and cytochemical studies of the spermatozoa of Acrosternum aseadum (Hemiptera: Pentatomidae) after copulation.

Beside the morphofunctional modifications undergone during spermiogenesis, the spermatozoon could undergo other modifications after copulation. Since no structural modification occurs in the spermatozoon of Acrosternum aseadum after copulation, we used cytochemical studies to show the enzymatic activities variations of acid phosphatase, thiamine pyrophosphatase, glucose-6-phosphatase and cytochrome C oxidase, when the spermatozoon passes through the spermatheca. The enzymatic activity, few hours after copulation, is strong and specifically located. However, 40 h after copulation, there is considerable loss of enzymatic activity, with the exception of thiamine pyrophosphatase, which shows the same activity. This result indicates that the spermatozoon of A. aseadum undergoes physiological modifications when passing through the spermatheca and that these modifications may be involved with survival in this organ as well as with the fertilization process.

Acid Phosphatase↗
PubMed1999

Modulation of glucose-induced insulin secretion by cytosolic redox state in clonal beta-cells.

Nutrient stimulation of pancreatic beta-cells increases the cellular reduced pyridine nucleotide content, but the specific role of cytosolic redox state in glucose-induced insulin release (GIIR) remains undetermined. The role of cytosolic redox state has been assessed (as reflected by the lactate/pyruvate ratio) in nutrient- and non-nutrient-induced insulin release using a recently established glucose-sensitive clonal beta-cell line (BRIN-BD11). Long-term exposure to the NAD+ precursor vitamin nicotinic acid (NA, 100 microM) was used to promote a more oxidized state in the cytosol. Glucose (2-16 mM) evoked a dose-dependent rise in the cytosolic NADH/NAD+ ratio which was linearly related to the extent of GIIR. NA suppressed the glucose-induced rise in the NADH/NAD+ ratio and concomitantly reduced GIIR by 44%. It also inhibited, by 47%, the average glucose-induced rise in cytosolic free Ca2+ concentration ([Ca2+]i, assessed by fura-2 microfluorometry from single cells). The latter effect was not accounted for by a reduction in the activity of voltage-sensitive Ca2+ channels, inasmuch as both high K+- and tolbutamide-induced [Ca2+]i rises remained insensitive to NA exposure. NA did not affect insulin release evoked by any of the depolarizing agents, indicating that steps in the stimulus-secretion coupling cascade distal to Ca2+ influx are insensitive to changes in the cytosolic redox state. It is concluded that GIIR is partially controlled by the cytosolic redox state. Moreover, the impairment in GIIR, caused by a shift toward a more oxidized state in the cytosol, originates from an attenuated [Ca2+]i response. The latter is likely mediated by the influence of cytosolic redox state on specific metabolic pathways (NADH shuttle systems and/or the malonyl-CoA pathway), leading ultimately to enhancement of the activity of ATP-sensitive K+ channels.

Animals↗
PubMed1999

An ultrastructural study of spermiogenesis in three species of Physalaemus (Anura, Leptodactylidae).

Ultrastructural aspects of spermiogenesis and testicular spermatozoa of three anuran amphibians, Physalaemus biligonigerus, P. fuscomaculatus and P. gracilis, were investigated by electron microscopy. The nuclei, middle pieces and tails of the three species were similar. In all cases, the nuclei were elongated and the acrosome consisted of a cone-shaped cap. The connecting pieces located in the flagellum implantation zone had transverse striations. The tails had a 9 + 2 axial filament pattern, a juxtaxonemal fiber and an undulating membrane. In contrast to other Leptodactylidae spermatozoa, no axial rod were observed in these Physalaemus species.

Animals↗
PubMed1998

Periodontology's application of multimedia.

This article makes an approach about the use of multimedia resources applied in odontology computer science educational, especially in periodontia. It highlights the importance of the use of these resources, as well as the one that than there is of more recent in this area.

Computer-Assisted Instruction↗
PubMed1998

Cytochemical localization of phosphatases in the germ- and Sertoli cells of Odontophrynus cultripes (Amphibia, Anura, Leptodactylidae).

Ultrastructural cytochemical techniques were used for the localization of phosphatases in spermatid and spermatozoon, as well as in Sertoli cells of Odontophrynus cultripes (Amphibia, Anura, Leptodactylidae). Acid phosphatase was found in the acrosome. Thiamine pyrophosphatase was observed in the Golgi cisternae and in the tail spermatozoon surface. Glucose-6-phosphatase was located in the membrane complex of the acrosomal region. Already, in the Sertoli cells acid phosphatase was located in the lysosomes and glucose-6-phosphatase was observed in association with the endoplasmic reticulum and Golgi complex. These observations support the idea that various phosphatases may play some role in spermatid differentiation and in the interactions germ cells--Sertoli cells during spermiogenesis process.

Acrosome↗
PubMed1998

Spermiogenesis in phytophagous bug (Hemiptera, Pentatomidae): an ultrastructural study.

Ultrastructural and immunocytochemical studies were carried out on spermiogenesis in two species of phytophagous bug, Acrosternum aseadum and Nezara viridula. The nucleus development involved changes in the shape and in the degree of chromatin condensation, with specific aggregation patterns of DNA-histone complex occurring during this process. The acrosome showed a three-layered and the acrosomal content a paracrystalline arrangement. The axoneme presented a 9 + 9 + 2 microtubule pattern and bridges occurred between axonemal microtubules 1 and 5, and mitochondrial derivatives. Two paracrystalline structures embedded in amorphous regions were observed in the mitochondrial derivatives. The use of the negative staining technique shows a zig-zag profile in the mitochondria due to infolding to the cristae, regularly spaced with approximately 40 nm. An electron dense rod was observed near the centriolar adjunct; it presented labelling for tubulin suggesting that this structure may be involved in the microtubule organization during spermiogenesis of these insects.

Animals↗