Clustering of Schistosoma mansoni mRNA sequences and analysis of the most transcribed genes: implications in metabolism and biology of different developmental stages

The study of the Schistosoma mansoni genome, one of the etiologic agents of human schistosomiasis, is essential for a better understanding of the biology and development of this parasite. In order to get an overview of all S. mansoni catalogued gene sequences, we performed a clustering analysis of the parasite mRNA sequences available in public databases. This was made using softwares PHRAP and CAP3. The consensus sequences, generated after the alignment of cluster constituent sequences, allowed the identification by database homology searches of the most expressed genes in the worm. We analyzed these genes and looked for a correlation between their high expression and parasite metabolism and biology. We observed that the majority of these genes is related to the maintenance of basic cell functions, encoding genes whose products are related to the cytoskeleton, intracellular transport and energy metabolism. Evidences are presented here that genes for aerobic energy metabolism are expressed in all the developmental stages analyzed. Some of the most expressed genes could not be identified by homology searches and may have some specific functions in the parasite.

Basic biology of Pneumocystis carinii: a mini review

Basic aspects of cell biology of Pneumocystis carinii are reviewed with major emphasis on its life cycle and the structural organization of the trophozoites and cyst forms. Initially considered as a protozoan it is now established that Pneumocystis belongs to the Fungi Kingdom. Its life cycle includes two basic forms: (a) trophozoites, which are haploid cells that divide by binary fission and may conjugate with each other forming an early procyst and (b) cysts where division takes place through a meiotic process with the formation of eight nuclei followed by cytoplasmic delimitation and formation of intracystic bodies which are subsequently released and transformed into trophozoites. Basic aspects of the structure of the two developmental stages of P. carinii are reviewed.

Brazilian contribution for a better knowledge on the biology of Toxoplasma gondii

Historically, scientists in Brazil has significantly contributed to the biology, cultivation and structural organization of the pathogenic protozoan Toxoplasma gondiiand its interaction with host cells, starting with the description of the protozoan by Splendore in 1908. The intracellular and extracellular corpuscoli observed in rabbits, corresponded to what we now as tachyzoites. Later on, a pioneering method to grow T. gondii in tissue cultures was developed by Guimarães and Meyer, 1942. They also observed for the first time T. gondii by transmission electron microscopy and made the initial description of the cytoskeleton of T. gondii by observing negatively stained cells. In the 1980's, the relation of the cytoskeleton with the sub-pellicular microtubules was reveled by freeze-fracture. More recently, several Brazilian groups have analyzed in detail basic aspects of the early interaction of the protozoan with the host cell, such as the role of protein phosphorylation, transfer of host cell surface components to the protozoan and genesis and organization of the parasitophorous vacuole. Tachyzoites strategically inhibit nitric oxide production during active invasion of activated macrophages. In vitro studies on the sexual cycle of T. gondii using primary cultures of cat enterocytes and the egress from host cells are being carried out. Perspectives are that the contribution of Brazilian science to the knowledge on T. gondii biology will continue to flourish in years to come.

Nesting biology of the trap-nesting Neotropical wasp Trypoxylo n(Trypargilum) aurifrons Shuckard (Hymenoptera, Crabronidae)

The present study was carried out in three localities of the state of São Paulo, Brazil: Araras (Dec/03-Dec/06), São Carlos (Nov/04-Nov/06) and Rifaina (Jul/04-Dec/06). Trap-nests were distributed among sites in the sampling areas and were collected every 35 days. Data from 295 nests indicate that T. aurifrons is a multivoltine species, with higher rates of nest building and cell production in the warm, rainy season. The trap-nests used by the females ranged from 117 to 467 mm in length and 3.1 to 16.6 mm in diameter. All nests showed deep plugs and a vestibular cell was found in 37% of the complete nests. The number of cells per nest ranged from one to 12. Females were larger than males, emerged from longer cells and their cocoons were significantly larger. A secondary 1:1 sex ratio was found in Araras and Rifaina. No correlation was observed between the diameter of the trap-nest and sex ratio. Males were usually oviposited in the first brood cells. Male and female developmental time from egg to adult was longer in the cold, dry season. Trypoxylon aurifrons provisioned their nests mainly with orb-spiders from the family Araneidae. The most important mortality factor was the death of immature forms, probably due to development failure. The most important parasitoid was Melittobia sp.

Reproductive biology and cytology of Hypericum brasiliense Choisy (Hypericaceae)

This is the first study of reproductive biology and cytology carried out with Hypericum brasiliense, a species with medicinal properties and potential agronomic interest. Three populations of H. brasiliense collected at Southeastern Brazil were studied. The results indicate that H. brasiliense is preferentially allogamous, self-compatible, facultative apomitic and anemophilous. Male sterility was observed in about 50% of individuals from the three populations. Anatomical studies evidenced structural abnormalities in anthers of male sterile flowers, showing enlarged tapetal cells and thick secretion deposits on the tapetal cell surfaces that may cause nutritional deficit for pollen mother cells. In cytogenetic studies several haploid chromosome numbers were observed like n = 4, 8, 9, 11, 16 and 17, including the presence of multivalents and micronuclei in tetrads, indicating the occurrence of abnormalities in the meiotic process of H. brasiliense. Despite these meiotic abnormalities the pollen viability and in vitro pollen germination rate observed in fertile flowers may be considered high. The diploid chromosome number 2n = 16 was observed, and the chromosomes in metaphase were small and similar. Fluorochrome staining techniques using DAPI and CMA3 were applied, with no positive bands observed.

Molecular biology of the kallikrein-kinin system: from structure to function

The participation of the kallikrein-kinin system, comprising the serine proteases kallikreins, the protein substrates kininogens and the effective peptides kinins, in some pathological processes like hypertension and cardiovascular diseases is still a matter of controversy. The use of different experimental set-ups in concert with the development of potent and specific inhibitors and antagonists for the system has highlighted its importance but the results still lack conclusivity. Over the last few years, transgenic and gene-targeting technologies associated with molecular biology tools have provided specific information about the elusive role of the kallikrein-kinin system in the control of blood pressure and electrolyte homeostasis. cDNA and genomic sequences for kinin receptors B2 and B1 from different species were isolated and shown to encode G-protein-coupled receptors and the structure and pharmacology of the receptors were characterized. Transgenic animals expressing an overactive kallikrein-kinin system were established to study the cardiovascular effects of these alterations and the results of these investigations further corroborate the importance of this system in the maintenance of normal blood pressure. Knockout animals for B2 and B1 receptors are available and their analysis also points to the role of these receptors in cardiovascular regulation and inflammatory processes. In this paper the most recent and relevant genetic animal models developed for the study of the kallikrein-kinin system are reviewed, and the advances they brought to the understanding of the biological role of this system are discussed.

Biology and clinical utilization of mesenchymal progenitor cells

Within the complex cellular arrangement found in the bone marrow stroma there exists a subset of nonhematopoietic cells referred to as mesenchymal progenitor cells (MPC). These cells can be expanded ex vivo and induced, either in vitro or in vivo, to terminally differentiate into at least seven types of cells: osteocytes, chondrocytes, adipocytes, tenocytes, myotubes, astrocytes and hematopoietic-supporting stroma. This broad multipotentiality, the feasibility to obtain MPC from bone marrow, cord and peripheral blood and their transplantability support the impact that the use of MPC will have in clinical settings. However, a number of fundamental questions about the cellular and molecular biology of MPC still need to be resolved before these cells can be used for safe and effective cell and gene therapies intended to replace, repair or enhance the physiological function of the mesenchymal and/or hematopoietic systems.

Indirect effect of neem oil on Podisus nigrispinus (Hemiptera, Pentatomidae): biology and predatory capacity

This study evaluated the effects on the development and predatory capacity of Podisus nigrispinus fed on Spodoptera frugiperda that have ingested different concentrations of neem oil. The predatory capacity of Podisus nigrispinus was assessed, separating nymphs (fourth instar) and adults (males and females). The treatments consisted of S. frugiperda larvae reared in neem oil aqueous solutions (0.077, 0.359 and 0.599%), deltamethrin EC 25 (0.100%) and control arranged in a completely randomized design, with ten replicates. Insects were offered three larval densities (one, three and six), in the third or fourth instars. The predated larvae were examined at 24 and 48 hours after the beginning of the experiment. Biological parameters of Podisus nigrispinus were evaluated in groups of ten second-instar nymphs transferred to pots, in five replicates. Insects were offered 2-6 third and/or fourth-instar larvae reared in the same neem oil concentrations in a completely randomized design. The following parameters were evaluated: duration of each nymph stage (days), nymph mortality (%), weight of fifth-instar nymphs (mg), sex ratio, weight of males and females (mg) and longevity of unfed adults (days). The predatory capacity of nymphs and adults of Podisus nigrispinus was influenced by the neem oil at the concentrations of 0.359% and 0.599% in the highest density. The concentration of 0.359% lengthened the nymphal stage and the concentration of 0.599% reduced the weight of males.

New Strategies on Molecular Biology Applied to Microbial Systematics

Systematics is the study of diversity of the organisms and their relationships comprising classification, nomenclature and identification. The term classification or taxonomy means the arrangement of the organisms in groups (rate) and the nomenclature is the attribution of correct international scientific names to organisms and identification is the inclusion of unknown strains in groups derived from classification. Therefore, classification for a stable nomenclature and a perfect identification are required previously. The beginning of the new bacterial systematics era can be remembered by the introduction and application of new taxonomic concepts and techniques, from the 50’s and 60’s. Important progress were achieved using numerical taxonomy and molecular taxonomy. Molecular taxonomy, brought into effect after the emergence of the Molecular Biology resources, provided knowledge that comprises systematics of bacteria, in which occurs great evolutionary interest, or where is observed the necessity of eliminating any environmental interference. When you study the composition and disposition of nucleotides in certain portions of the genetic material, you study searching their genome, much less susceptible to environmental alterations than proteins, codified based on it. In the molecular taxonomy, you can research both DNA and RNA, and the main techniques that have been used in the systematics comprise the build of restriction maps, DNA-DNA hybridization, DNA-RNA hybridization, sequencing of DNA sequencing of sub-units 16S and 23S of rRNA, RAPD, RFLP, PFGE etc. Techniques such as base sequencing, though they are extremely sensible and greatly precise, are relatively onerous and impracticable to the great majority of the bacterial taxonomy laboratories. Several specialized techniques have been applied to taxonomic studies of microorganisms. In the last years, these have included preliminary electrophoretic analysis of soluble proteins and isoenzymes, and subsequently determination of deoxyribonucleic acid base composition and assessment of base sequence homology by means of DNA-RNA hybrid experiments beside others. These various techniques, as expected, have generally indicated a lack of taxonomic information in microbial systematics. There are numberless techniques and methodologies that make bacteria identification and classification study possible, part of them described here, allowing establish different degrees of subspecific and interspecific similarity through phenetic-genetic polymorphism analysis. However, was pointed out the necessity of using more than one technique for better establish similarity degrees within microorganisms. Obtaining data resulting from application of a sole technique isolatedly may not provide significant information from Bacterial Systematics viewpoint