Patologia do Coração na AIDS. Estudo de 73 Necropsias Consecutivas

Purpose - To study the incidence and the etiology of the cardiac lesions in AIDS patients. Methods - The autopsy protocols and the filled slides of the heart from 73 consecutive AIDS patients were reviewed. There were, at least, 2 slides of each heart stained by haematoxylin-eosin; when indicated, Ziehl-Nielsen, Gram and Gomori Grocott stains were used. Results - No cause of death was assigned to the heart. There was involvement of the heart in 66 (90%) cases. Marked atrophy of cardiac fibers with or without lipomatosis was observed in 38 patients. Interstitial infiltrates of myocardium were present in 38 necropsies and in 13 of these cases a probable pathogen was demonstrated: cryptococcus neoforms in three cases and mycobacteria tuberculosis, atypical mycobacteria, toxoplasma gondii, trypanosoma cruzi and cytomegalovirus in two cases each. Bacterial endocarditis was found in 4 autopsies and Kaposi sarcome in one. The pericardium was involved in 22 cases; in 12 there was only non specific mononuclear infiltration. Conclusion - Autopsy examination of the heart from AIDS patients revealed frequent pathologic involvement.

Solid mass falling into the reservoirs shallow water equations validity from the engineering point of view

Water waves generated by a solid mass is a complex phenomenon discussed in this paper by numerical and experimental approaches. A model based on shallow water equations with shocks (Saint Venant) has developed. It can reproduce the amplitude and the energy of the wave quite well, but because it consistently generates a hydraulic jump, it is able to reproduce the profile, in the case of high relative thickness of slide, but in the case of small relative thickness it is unable to reproduce the amplitude of the wave. As the momentum conservation is not verified during the phase of wave creation, a second technique based on discharge transfer coefficient α, is introduced at the zone of impact. Numerical tests have been performed and validated this technique from the experimental results of the wave's height obtained in a flume.

Kariological study of five Arecoideae palmae species

Kariological data for 5 species of palm subfamily Arecoideae are presented based on somatic methapase chromosomes obtained from seeds germinated on Sphagnum, and using standard methodology for the slides preparation. From the kariogram of Aiphanes. acanthophylla (2n=30),A. caryotaefolia (2n=30), Syagrus coronata (2n=32), S. quinquifaria (2n=32) and Scheelea lauromuelleriana (2n=32) the relative centromere position (short arm x 100/total chromosome length) and the relative length of each chromosome (average chromosome length/total chromosome length) were calculated.

Methodology for kariological study of Brazilian Palms

In the laboratory of cytogenetics of the DBAA-UNESP we are studying the karyotipe of some Brazilian Palms. To determine the best protocol, methods of seed germination, inhibition of mitosis, time to pick up the roots and staining were analyzed. The results shown that the seed germination in sphagnum is effective to achieve good roots. The best time to collect the root tips is between 11 to 12 AM., when there are more cell metaphases. The inhibition of mitosis cycle at metaphases may be effective both with 8-hydroxiquinoleine (0,03% -5 hours) or with cold water (0°C - 18-20 hours). The staining with Giemsa 2% showed the best chromosome figures in the metaphases. Now, to get good metaphases slides the method in use in the lab is: 1) seed germination in sphagnum at room temperature and high humidity; 2) The major roots are cut maintaining at least 5 cm, because this technic allows new emergence of roots, increasing the number of roots collected per germinated seed, that is very important in some species with poor germination rates; 3) To get the mitosis inhibition we are using cold water (0°C) treatment for 18-20 hours, following the standard protocols for conservation and hydrolysis; or enzyme digestion with pectinasecellulase 4) the staining procedures are made using Giemsa 2%. The Brazilian palms species studied and their respective chromosome number were: Aiphanes acanthophylla (2n=30), A. caryotaefolia (2n=30), Syagrus quinquifaria (2n=32), S. coronata (2n=32), S. romanzoffiana (2n=32), Euterpe edulis (2n=36), E. oleracea (2n=36), Copernicia prunifera (2n=36), Scheelea lauromuelleriana (2n=32) and Bactris gasipaes (2n=30).

Nucleotide sequence of 5S rDNA and localization of the ribosomal RNA genes to metaphase chromosomes of the Tilapiine cichlid fish, Oreochromis niloticus

In this study, we report the cloning and nucleotide sequence of PCR-generated 5S rDNA from the Tilapiine cichlid fish, Oreochromis niloticus. Two types of 5S rDNA were detected that differed by insertions and/or deletions and base substitutions within the non-transcribed spacer (NTS). Two 5S rDNA loci were observed by fluorescent in situ hybridization (FISH) in metaphase spreads of tilapia chromosomes. FISH using an 18S rDNA probe and silver nitrate sequential staining of 5S-FISH slides showed three 18S rDNA loci that are not syntenic to the 5S rDNA loci.

An improved three-dimensional model for growth of oxide films induced by laser heating

In this paper we consider a three-dimensional heat diffusion model to explain the growth of oxide films which takes place when a laser beam is shined on and heats a metallic layer deposited on a glass substrate in a normal atmospheric environment. In particular, we apply this model to the experimental results obtained for the dependence of the oxide layer thickness on the laser density power for growth of TiO2 films grown on Ti-covered glass slides. We show that there is a very good agreement between the experimental results and the theoretical predictions from our proposed three-dimensional model, improving the results obtained with the one-dimensional heat diffusion model previously reported. Our theoretical results also show the occurrence of surface cooling between consecutive laser pulses, and that the oxide track surface profile closely follows the spatial laser profile indicating that heat diffusive effects can be neglected in the growth of oxide films by laser heating. © 2001 Elsevier Science B.V.

The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR)

The objective of this study was to compare the different methods of detecting Toxoplasma gondii in sheep tissue, tested serologically positive by the indirect immunofluorescent antibody test (IFAT). Brain, diaphragm, and blood samples were collected from 522 sheep slaughtered at the São Manuel abattoir, São Paulo State, Brazil. Brain and diaphragm samples from IFAT seropositive animals were digested by both trypsin and pepsin and then injected into mice. Part of the digested samples was used to prepare slides for Giemsa staining and in the polymerase chain reaction (PCR). Tissue fragments were fixed in formalin and examined using hematoxilin-eosin (HE). Forty of the sheep (7.7%) were IFAT positive. T. gondii was isolated in 23 (59.0%) of the 39 mice with pepsin-digested brain samples and in 27 (69.0%) of the 39 with trypsin-digested brain samples. Injection of diaphragm samples led to T. gondii isolation in 26 (66.7%) of the 39 pepsin-digested samples and 21 (53.8%) of the 39 trypsin-digested samples. Cytological and hystopathological examination of both brains and diaphragms was negative in all examined sheep. PCR was positive in 7 (17.9%) of the trypsin and 2 (5.1%) of the pepsin-digested samples, while 9 (23.1%) of the trypsin and 3 (7.7%) of the pepsin-digested samples showed T. gondii DNA. T. gondii isolation rate in mice (n = 34; 85.0%) was significantly higher than detection by PCR (n = 15; 37.5%). © 2001 Elsevier Science B.V.

Evolução no diagnóstico citológico do câncer de colo uterino - Novas tecnologias para o rastreamento citológico cervical

The cancer of the uterine cervix remains a serious public health problem in developing countries with low social-economic conditions. Papanicolaou smear has been used to screen women for cervical cancer since 1940, and it is widely recognized as an effective method for preventing cervical cancer. In Brazil, conventional Papanicolaou is the most common method used in diagnosis cytopathological routine, involving the morphological cell analysis by microscopic examination of cervical samples smeared on glass slides. On the other hand, the liquid cytology is an advantageous method whose specimen collected is put into a fluid alcohol based to guarantee the potential use of residual cellular material in cytobrush. Moreover, this method permits additional diagnostic studies, preserving the morphological cell features as the conventional one does. The aim of the present study was report the historical evolution of different cervical cytologic tests, emphasizing the advantages and disadvantages between conventional Papanicolaou method and new technologies in liquid based cytology.

Mutagenic potential evaluation of the water of a river that receives tannery effluent using the Allium cepa test system

Several studies are being conducted to assess the toxicity and cytotoxicity of water bodies receiving industrial and domestic effluents, using the Allium cepa test. To assess the toxicity and mutagenicity of water possibly contaminated with chromium, derived from tannery activities, seasonal water samplings were performed in 2001 and 2002 at five different sites along the Sapucaizinho river, Municipality of Patrocinio Paulista, State of São Paulo, Brazil. A. cepa seeds were used as the test material and were submitted to germination in waters from the different collection sites, in Milli-Q water (negative control) and in aqueous solution of chromium (positive control). For the determination of cell division rates and mitotic irregularities, slides were prepared with root tip cells according to the standard Feulgen methodology. The results showed that the collection sites most heavily compromised by chromium emission presented low mitotic indices and a higher frequency of mitotic changes such as irregular anaphases (disorganized, multipolar, laggard), cells with chromosomal adherences, cells with micronuclei, and binucleate and/or multinucleate cells. © 2004 The Japan Mendel Society.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Decrease of the DNA damage levels after sperm preparation by layering method

The aim of this prospective study was to determine the DNA fragmentation levels before and after sperm preparation by layering method. A total of 78 patients submitted to assisted reproduction technology (ART) for infertility treatment were evaluated. Ejaculated spermatozoa were obtained by masturbation on the day of ART procedure. The evaluation of DNA fragmentation was performed in the fresh semen and after preparation by a layering method, respectively. After washing with PBS, the sperm pellets were smears and then processed for the terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) assay that was performed using a Cell Death Detection Kit with tetramethylrhodamine-labelled dUTP. For quantitative evaluation, 200 spermatozoa in randomly selected areas on microscope slides were evaluated and the percentage of TUNEL positive spermatozoa was determined. If ≥20% of selected sperm were TUNEL positive, the exam was considered abnormal. The mean percentage of DNA sperm fragmentation before sperm preparation was 17±8.3% and after 7.8±6.5% (p<0.0001). The exam was considered normal in 49 patients before preparation and in 73 patients after (p<0.0001). The sperm preparation with a layering method for the ART procedure is effective to select sperm with a significant decrease of the DNA damage.

The effects of male age on sperm DNA damage in an infertile population

The objective was to investigate the influence of age on sperm DNA damage. Semen samples were collected from 508 men in an unselected group of couples attending infertility investigation and treatment. DNA fragmentation in spermatozoa was measured by TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay; at least 200 spermatozoa in randomly selected areas of microscope slides were evaluated using a fluorescent microscope and the percentage of TUNEL positive spermatozoa was determined. The number of cells with red fluorescence (TUNEL positive) was expressed as a percentage of the total sample [DNA fragmentation index (DFI)]. Age was treated as a continuous variable for regression and correlation analysis. The following male age groups were used: Group I: ≤35 years, Group II: 36-39 years, and Group III: ≥40 years. DFI was significantly lower in Group I than in Group II (P = 0.034) or III (P = 0.022). There was no difference in DFI between Groups II and III. In addition, regression analysis demonstrated a significant increase in sperm DFI with age (P = 0.02). TUNEL assay clearly demonstrates an increase in sperm DNA damage with age. © 2007 Published by Reproductive Healthcare Ltd.

Description and morphometry of the sting apparatus in the three worker casts of Acromyrmex subterraneus brunneus (Formicidae, Attini)

We studied the morphometry of the sting apparatus of Acromyrmex subterraneus brunneus workers by dissection and mounting of the structures on slides, and present here a brief description and illustrations. Sting apparatus examinations showed differences in size, but without morphological differences. The seven morphometric variables studied permitted grouping of the Ac. subterraneus brunneus workers into the following three groups: cluster 1 - consisting of 85, 39 large, 45 medium and 1 small workers; cluster 2 - consisting of 44 small workers, and cluster 3 - consisting of 6 large workers. This study demonstrated that all worker castes follow the same morphological pattern, although functional characteristics of the sting apparatus should be highlighted.

Cryptosporidium SP in HIV-infected individuals attending a Brazilian university hospital

The aim of the current work was to evaluate the occurrence of Cryptosporidium sp in AIDS patients in a region of São Paulo State, Brazil. Patients were divided into groups according to CD4+ T lymphocyte count and use of potent antiretroviral treatment. Two hundred and ten fecal samples from 105 patients were fixed in 10% formalin and subjected to centrifuge formol-ether sedimentation. Slides were stained with auramine and confirmed by modified Ziehl-Neelsen. Cryptosporidiosis occurrence was 10.5% with no relationship among gender, age or the presence of diarrhea. The number of oocysts in all samples was small, independent of CD4+ T lymphocyte count, HIV plasma viral load, and presence of diarrhea. These results may be due to the reduced prevalence of opportunistic infections in AIDS individuals after the advent of highly active antiretroviral therapy.

Optimal clone identifier for genomic shotgun libraries: OC Identifier tool

In DNA microarray experiments, the gene fragments that are spotted on the slides are usually obtained by the synthesis of specific oligonucleotides that are able to amplify genes through PCR. Shotgun library sequences are an alternative to synthesis of primers for the study of each gene in the genome. The possibility of putting thousands of gene sequences into a single slide allows the use of shotgun clones in order to proceed with microarray analysis without a completely sequenced genome. We developed an OC Identifier tool (optimal clone identifier for genomic shotgun libraries) for the identification of unique genes in shotgun libraries based on a partially sequenced genome; this allows simultaneous use of clones in projects such as transcriptome and phylogeny studies, using comparative genomic hybridization and genome assembly. The OC Identifier tool allows comparative genome analysis, biological databases, query language in relational databases, and provides bioinformatics tools to identify clones that contain unique genes as alternatives to primer synthesis. The OC Identifier allows analysis of clones during the sequencing phase, making it possible to select genes of interest for construction of a DNA microarray. ©FUNPEC-RP.