Proteins of Leishmania (Viannia) shawi confer protection associated with Th1 immune response and memory generation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spermatogenesis and nucleolar behavior in males of aquatic Heteroptera

Aspects of spermatogenesis and nucleolar behavior were analyzed in Brachymetra albinerva, Cylindrostethus palmaris, Halobatopsis platensis, Limnogonus aduncus (Gerridae), Martarega sp (Notonectidae), Rhagovelia whitei, and Rhagovelia sp (Veliidae). The testicles are rounded (Veliidae), elongated (Gerridae) or spiral (Notonectidae) and have a transparent membrane covering them. The complement chromosome was 2n = 23 (22A + X0, L. aduncus and Rhagovelia sp), 25 (24A + X0, B. albinerva and H. platensis), 26 (22A + 2m + XY, Martarega sp), 29 (28A + X0, C. palmaris), or 39 (38A + X0, R. whitei) chromosomes, and the only species with a different sex chromosome system was Martarega sp, which showed an XY system and m-chromosomes. The meiotic behavior of all species was similar: holocentric chromosomes and heteropyknotic material at prophase, interstitial and/or terminal chiasmata, and first reductional division for the autosomes and the reverse for the sex chromosomes. The only difference observed was related to the very large size of Martarega sp cells in all stages of spermatogenesis. With regard to nucleolar behavior, the species did not show differences, except for Martarega sp with larger nucleoli than the other species. The only species in which it was clearly possible to identify the nucleolar organizer region was L. aduncus, in the region of a terminal autosome. It was also confirmed that the telomeric associations do not occur at random. In the other species, specific staining was very discrete, and the nucleolar organizer region location was not at all evident.

Patterns of nucleolar activity during spermatogenesis of two triatomines, Panstrongylus megistus and P. herreri

The present work analyzed spermatogenesis in two species of triatomines (genus Panstrongylus) using silver-ion impregnation. The sex chromosomes of P. megistus and P. herreri had nucleolar organizing activity and became strongly impregnated during the phases of meiotic prophase I. Fragmentation of the nucleolus occurred in both species during the meiotic cycle. The nucleolar region could be observed up to diakinesis in meiotic prophase after which only nucleolar bodies and fragments were seen. Postmeiotic reactivation of rRNA synthesis occurred in these two species and was probably related to cell differentiation.

Invited review Giardia duodenalis: Inter-strain variability of proteins, antigens, proteases, isoenzymes and nucleic acids

Giardia duodenalis isolates from asymptomatic or symptomatic patients and from animals present similarities and differences in the protein composition, antigenic profile, pattern of proteases and isoenzymes, as well as in nucleic acids analysis. In the present overview, these differences and similarities are reviewed with emphasis in the host-parasite interplay and possible mechanisms of virulence of the protozoon.

Argyrophilic nucleolar organizer regions (AgNORs) in odontogenic myxoma (OM) and ameloblastic fibroma (AF)

Ten cases of odontogenic myxoma (OM) and six cases of ameloblastic fibroma (AF) were subjected to comparative analysis by the AgNOR technique, in order to determine a possible difference in cell proliferation index between these lesions. The mean AgNOR number of the mesenchymal component of AF was compared with its epithelial component and the difference was not found to be statistically significant. The mean AgNOR index of the AF group was significantly higher than that of the OM group. Moreover, the mesenchymal component of AF demonstrated increased AgNOR numbers compared with that of OM (P<0.05). These results suggest that the epithelial and mesenchymal components of AF may have similar cell proliferative activity. However, the cell proliferative index of this lesion seems to be higher than that of OM.

Nucleolar changes in malpighian tubules cells of Rhodnius prolixus during starvation

This study focused on nuclear and nucleolar area changes in Malpighian tubule cells of Rhodnius prolixus, an hematophagous insect, vector of Chagas' disease. Male and female adult insects were dissected after a 28-day starvation period, as well as insects which had been fed again after a 30-day starvation period. Malpighian tubules were fixed and silver stained. In both, males and females, nucleolar fusions and regions of nucleolar corpuscles were observed to be differentially impregnated by silver during feeding stress. In the males and females that were fed again, nucleolar corpuscles were partially fusioned, indicating a slight recovery upon the 30-day starvation period. The changes observed in the nucleolar phenotype and in both nuclear and nucleolar areas indicated that, as a result of stress, a more intense, compensatory activity occurred to supply the metabolic rate. The mechanism of this phenomenon may be associated to the decondensation and activation of chromatin that carries rDNA in order to increase rRNA transcription, and,consequently, protein synthesis.

Juvenile hormone promotes changes in the expression of hypopharyngeal gland proteins of worker Apis mellifera (Hymenoptera: Apidae)

The present investigation compares the protein electrophoreses profiles of the hypopharyngeal glands of 12 and 25 day old Apis mellifera workers, some of which were experimentally treated with an analogue of juvenile hormone in the moment of the emergence while others were not treated. According to the evaluation of the presented variations by four main bands, it is concluded that the analogue juvenile hormone changes the glandular genetic expression pattern, promoting the disappearance of two from the four main bands in 25 day old workers. The effect of this hormone is discussed as an hypopharyngeal maturation inductor, in synergetic action with the bee age acting early in the glandular cycle.

Influence of the application of juvenile hormone on the haemolymph total proteins and electrophoretic pattern of worker larvae of Apis mellifera

The aim of the present work was to verify the influence of the juvenile hormone (JH) applied on worker larvae of Apis mellifera 2 to 5 days old over the haemolymph total protein and electrophoretic pattern. Each larvae received topical applications of 1 ml of a solution of JH in hexane (1 μg/ml) on their 2 nd, 3 rd 4 th and 5 th day after hatching and had the amount and electrophoretic pattern of proteins from the haemolymph analyzed during the remaining days of their life. As a control, haemolymph of larvae of the same age that did not receive any kind of treatment was analyzed. The results show that the application of JH on larvae 3 or more days old affect the amount and electrophoretic pattern of the proteins, with this effect lasting through the subsequent days.

The effect of glycerol on signal supression during electrospray ionization analysis of proteins

The effect of salts, detergents and chaotropic agents on mass spectrometric analysis are relatively well understood, mainly due to their actions decreasing the performance of ESI interface in mass spectrometric analysis. However, there are few studies in the literature characterizing the effect of protein stabilization by glycerol, followed in some circumstances by the suppression of protein signal when ESI interface is used. The aim of the present research was to investigate in details the mass spectrometric behavior of some proteins in presence of high levels of glycerol during ESI-MS analysis. Thus, horse heart myoglobin and chicken ovalbumin were used as standard proteins. It was demonstrated that the presence of 1% (v/v) glycerol suppressed the signal of these proteins during the ESI-MS analysis, even when the sample nozzle potential was scanned from 28 to 80 V. However, when the glycerol concentration was decreased to 0.5% (v/v) and the sample cone voltage adjusted to 50 V, a perfect envelope of peaks was observed, allowing the spectrum deconvolution and the molecular mass determination with mass accuracy lower than 0.01% in each situation. A molecular explanation for this suppressive effect and for the analytical overcoming of this difficult is proposed.

Description of the nucleolar activity and karyotype in germinative cell lines of Rhodnius domesticus (Triatominae, Heteroptera)

The purpose of this work was to study the karyotype, spermatogenesis and nucleolar activity at meiosis, in the species Rhodnius domesticus (Heteroptera, Triatominae). The testicular tubules were cytologically prepared by the conventional method of cell crushing and subsequent application of cytogenetic staining techniques with lacto-acetic orcein and silver-ion impregnation. The species under study presented karyotype 2n= 20A+XY, the modal number of the subfamily Triatominae. The chromosomes presented no primary constriction and were therefore characterized as holocentric. It was observed that the sex chromosomes sometimes were located at the periphery, close to the ring formed by autosomes, at first meiotic division. At metaphases II, sex chromosomes were positioned in the center of the autosomal ring, thus evidencing a postreductional behavior. These same chromosomes showed late migration at anaphases and were clearly impregnated with silver-ions, suggesting they bore Nucleolar Organizer Regions. Dispersed nucleolar corpuscles in cytoplasm until telophase II and small dots in spermatids strongly impregnated with silver, could be seen. Thus, it may be inferred that, in triatomines, the nucleolus does not completely disappear but remains in the form of small corpuscles that have a role in cell differentiation.

Cytogenetic study on three species of the genus Triatoma (Heteroptera: Reduviidae) with emphasis on nucleolar organizer regions

The use of banding techniques allows the recognition of chromosomal pairs and karyotypical arrangements. However, its application in Heteroptera holocentric chromosomes is limited. Thus, little is known about their structure, specially their Nucleolar Organizer Regions (NORs). A comparative analysis of the nucleolar characteristics present during spermatogenesis in Triatoma platensis, Triatoma protacta and Triatoma tibiamaculata seems to indicate that in this group of insects nucleolar fragmentation occurs after prophase I. The study of chromosomal structure of these triatomines indicates that NORs are located at some telomeric and interstitial autosome regions and at sexual chromosomes (X/X1X2).

Solubilization of proteins from human lymph node tissue and two-dimensional gel storage.

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT (dithiothreitol) and 0.2% carrier ampholytes; (b) 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate), 40 mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7 M urea, 2 M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

New evidence for nucleolar dominance in hybrids of Drosophila arizonae and Drosophila mulleri

Drosophila mulleri (MU) and D. arizonae (AR) are cryptic species of the mulleri complex, mulleri subgroup, repleta group. Earlier cytogenetic studies revealed that these species have different regulatory mechanisms of nucleolar organizing activity. In these species, nucleolar organizing regions are found in both the X chromosome and the microchromosome. In the salivary glands of hybrids between MU females and AR males, there is an interspecific dominance of the regulatory system of the D. arizonae nucleolar organizer involving, in males, amplification and activation of the nucleolar organizer from the microchromosome. The authors who reported these findings obtained hybrids only in that cross-direction. More recently, hybrids in the opposite direction, i.e., between MU males and AR females, have been obtained. The purpose of the present study was to evaluate, in these hybrids, the association of the nucleoli with the chromosomes inherited from parental species in order to cytogenetically confirm the dominance patterns previously described. Our results support the proposed dominance of the AR nucleolar organizer activity over that of MU, regardless of cross-direction. ©FUNPEC-RP.