Pattern of nucleolar activity during spermiogenesis in triatomines (Heteroptera, Reduviidae) as analysed by silver staining

Nucleoli are the sites of biosynthesis of ribosomal precursors. In this work the nucleolar activity at interphase and the meiotic cells of the testis in five species of triatomines were analysed by means of silver staining. Several nucleolar blocks in the polyploid nuclei of testicular tubules were observed, whereas only one nucleolar body could be seen in the spermatogonial nuclei of all five species. A single nucleolar body was evident in the 'confused stage' of Triatoma brasiliensis, T. delpontei, T. lecticularia and T. rubrovaria, while T. sordida presented two nucleolar dots. The existence of small, silver-stained dots in some metaphase I chromosomes of T. brasiliensis and T. sordida is reported. The number of nucleolar dots present in spermatids of each species varied within and among species. It is suggested that in addition to providing information on rRNA biosynthesis, studies of nucleolar organizing activity can also be important sources of data on differentiation patterns and species development.

The applicability of hematoxylin-eosin staining plus fluorescence or confocal laser scanning microscopy to the study of elastic fibers in cartilages

This study focuses on the use of hemotoxylin-eosin staining plus fluorescence microscopy for the investigation of elastic fibers in some elastic cartilages. We have observed that elastic fibers are consistently imaged by the proposed procedure and the resolution attained is similar to that obtained with the classical Weigert's fuchsin-resorcin. The results also demonstrate that elastin autofluorescence gives little or no contribution to the final fluorescence and that the use of the confocal laser scanning microscope adds to the resolution, permits the use of thicker sections and reveals of minute structural features. We conclude that this is a relevant tool in elastin research.

Picrosirius-polarization staining method as an efficient histopathological tool for collagenolysis detectin in vesical prolapse lesions

The Picrosirius-polarization method has been indicated as a selective histochemical stain for collagen detection in tissue sections. This method can also be of value for studying collagen degradation given that, under polarized light, collagen displays birefringence due to its molecular order. The aim of this study is to highlight this staining method as an additional instrument for a rapid and excellent confirmatory diagnosis of the presence of collagenolysis in connective tissue in the vaginal wall with vesical prolapse lesion, in tissue sections. Dramatic changes in collagen morphology were found in vaginal mucosa in vesical prolapse disorder: they were weakly stained by Sirius red and under polarized light appeared as thin, pale (weakly birefringent), greenish, and with fibers more scattered, while the histoarchitecture of the organ showed a disrupted appearance. Thus, in the present study, we showed in vaginal mucosa in the vesicle prolapse that corroded collagenous framework appears as fragmentary and irregularly separated collagenous structures, that are weakly birefringent, corresponding to a molecular disorganization of these fibers caused by collagenolysis. (C) 2006 Elsevier Ltd. All rights reserved.

A COMPARATIVE-STUDY OF 4 DIFFERENT STAINING METHODS FOR ESTIMATION OF LIVE YEAST FORM CELLS OF PARACOCCIDIOIDES-BRASILIENSIS

A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis was carried out. The staining methods used were fluorescent staining, vital dye exclusion tests with erythrosin B and by Janus green and lactophenol cotton blue staining. Colony forming units (cfu) of the yeast form of eight P. brasiliensis isolates on brain heart infusion agar (BHIA) supplemented with 4% horse serum plus 5% P. brasiliensis cell extract (BHIA + HS + EXT) were examined for reliability of staining in determining the number of live fungal units in eight different isolates. Cfu on BHIA + HS + EXT plates showed an excellent plating efficiency over 96% in all isolates tested. The percentage of the live cells indicated by fluorescent staining (FL) or vital dye exclusion test with erythrosin B (EB) or Janus green (JG-1) was lower than that of cfu. By contrast, the percentage due to modified dye exclusion test with Janus green (JG-2) and that due to lactophenol cotton blue staining (LPCB) showed a close correration to that of cfu. Our results indicate that the modified dye exclusion test with Janus green and lactophenol cotton blue staining are useful for estimating cell viability of yeast form cells of P. brasiliensis.

DISTRIBUTION OF THE NUMBER OF EURYTREMA SP EGGS PER GRAM OF FECES IN NATURALLY INFECTED CATTLE

Eurytrema sp. egg counts (epg) in the feces of naturally infected cattle were performed and the technique employed showed 94.2% probability of detecting positive cases of the infection with a single examination independently of the host parasite burden. It was also demonstrated that the epg of Eurytrema sp. follows a negative binomial distribution model and is characterized by its small magnitude.

Complete staining of nerve fiber and myoneural junctions with acetylthiocholine and silver

We describe a combined stain for simultaneous demonstration of the preterminal axons and cholinesterase activity at myoneural junctions of mammalian muscles. This technique employs acetylthiocholine iodide as the substrate for cholinesterase activity and silver nitrate impregnation of preterminal axons. The procedure is rapid, simple and uses fresh muscles. Intramuscular nerves, preterminal axons and myoneural junctions are stained simultaneously brown or black with minimal background staining of connective tissue and muscle fibers.

Infecção experimental pelo Encephalitozoon cuniculi em camundongos imunossuprimidos com dexametasona

Objective Microsporidian Encephalitozoon cuniculi has been recognized as an opportunistic pathogen in immunosuppressed individuals, such as AIDS patients. The objective of the study was to develop pharmacologically immunosuppressed animals as a model of the natural occurring E. cuniculi infection.Methods Distint groups of adult Balb-C mice were immunosuppressed with different doses of dexamethasone (Dx, 3 or 5 mg/kg/day, intraperitoneal route - IP) and inoculated with E. cuniculi spores by IP route intraperitoneally. Control groups (inoculated animals but non-immunosuppressed and non-inoculated animals but immunosuppressed) were also used. The spores of E. cuniculi were previously cultivated in MDCK cells. The animals were sacrificed and necropsied at 7, 14, 21, 28 and 35 days post-inoculation. Tissue fragments were collected and processed for light microscopy studies, using Gram-chromotrope and hematoxylin-eosin staining techniques.Results In all immunosupressed and inoculated inoculated immunosuppressed mice,specially in those that received 5 mg/kg/day of dexamethasone, the most prominent necropsy findings were hepatomegaly and splenomegaly. The experimental inoculation resulted in a disseminated non-lethal infection, characterized by granulomatous lesions in several organs (liver lungs, kidneys, gut and brain) but notably in the hepatic tissue. Spores of E. cuniculi were only seen in few animals treated with 5 mg/kg/day of Dx at 35 days post-infection.Conclusions Microsporidiosis in Dx-immunosuppressed mice provides a useful model for studies of the microsporidial infection, resembling that one naturally occurring in immunodeficient individuals with AIDS.

Chromosomal studies on five species of the genus Leptodactylus Fitzinger, 1826 (Amphibia, Anura) using differential staining

Cytogenetic studies were carried out on five species of Leptodactylus, namely L. fuscus, L, notoaktites, L. labyrinthicus, L. ocellatus, and L. podicipinus, after standard staining, Ag-NOR and C-banding as well as BrdU incorporation for three of them. The species had 2n = 22 chromosomes and two basic karyotype patterns. Chromosome 8 was a marker bearing a secondary constriction. In all species, this secondary constriction corresponded to the Ag-NOR site. The species had centromeric C-bands in all chromosomes of the complement, but some interstitial or telomeric bands seemed to differentiate some karyotypes, either at the species or the population level. In L. ocellatus, the C-banding pattern confirmed the occurrence of a heteromorphic pericentric inversion in chromosome 8 in specimens from one of the populations. The BrdU incorporation technique showed no detectable difference in the replication patterns of the major bands in the chromosomes of L. noroaktites, L. labyrinthicus, and L. ocellatus.

Evaluation of the Possibility of Removing Staining by Repolishing Composite Resins Submitted to Artificial Aging

This in vitro research verified the possibility of eliminating staining caused by coffee and red wine in five composite resins, after being submitted to thermal cycling. Thirty-six specimens were prepared and immersed in water at 37 degrees C for 24 hours. After polishing, specimen color was measured in a spectrophotometer Cintra 10 UV (Visible Spectrometer, GBC, Braeside, VIC, Australia). All specimens were submitted to thermal cycling at temperatures of 5 and 55 degrees C with a dwell time of 1 minute, for 1,000 cycles in a 75% ethanol/water solution. After thermal cycling, the specimens were immersed in water at 37 degrees C until 7 days had elapsed from the time the specimens were prepared. All specimens were then taken to the spectrophotometer for color measurement. The specimens were divided into three groups (N = 12): distilled water (control), coffee, and red wine. For the staining process to occur on only one surface, all the sides, except one, of the surfaces were isolated with white wax. The specimens were immersed in one of the solutions at 37 degrees C for 14 days. The specimens were dried and taken to the spectrophotometer for color measurement. After this, the specimens were submitted to 20 mu m wear three times, and the color was measured after each one of the wear procedures. Calculation of the color difference was made using CIEDE2000 formula. According to the methodology used in this research, it was concluded that the staining caused by coffee and red wine was superficial and one wear of 20 mu m was sufficient to remove the discoloration.

Characteristics of gram-stained cervical smear from patients with Chlamydia trachomatis infection.

Cervical discharges from 142 women attending the Public Gynecologic Service of Araraqura (SESA), Brazil were cultured for Chlamydia trachomatis. Gram-smears and plating on semiquantitative sheep blood agar and chocolate agar were also carried out. An isolation rate of 18% was reported. The presence of purulent cervical secretion was observed in 8 (32%) out of 25 women. It was also observed that a substantial proportion of culture-positive women had no symptoms. Our data demonstrate that screening tests should be based on specific diagnostic techniques for Chlamydia trachomatis since the majority of infected women we examined were asymptomatic.

Cytogenetic analysis of Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) with Xyp sex determination system using standard staining, C-bands, NOR and synaptonemal complex microspreading techniques

The mitotic and meiotic chromosomes of the beetles Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) were analysed using standard staining, C-banding and silver impregnation techniques. We determine the diploid and haploid chromosome numbers, the sex determination system and describe the chromosomal morphology, the C-banding pattern and the chromosome(s) bearing NORs (nucleolar organizer regions). Both species shown 2n = 20 chromosomes, the chromosomal meioformula 9 + Xyp, and regular chromosome segregation during anaphases I and II. The chromosomes of E. atomaria are basically metacentric or submetacentric and P. dermestoides chromosomes are submetacentric or subtelocentric. In both beetles the constitutive heterochromatin is located in the pericentromeric region in all autosomes and in the Xp chromosome; additional C-bands were observed in telomeric region of the short arm in some autosomes in P. dermestoides. The yp chromosome did not show typical C-bands in these species. As for the synaptonemal complex, the nucleolar material is associated to the 7th bivalent in E. atomaria and 3rd and 7th bivalents in P. dermestoides. Strong silver impregnated material was observed in association with Xyp in light and electron microscopy preparations in these species and this material was interpreted to be related to nucleolar material.