The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.

Central nervous system of Rhipicephalus sanguineus ticks (Acari: Ixodidae): an ultrastructural study

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

Inner satellites of Neptune: I the disturbing function

In this work, we focus our attention to the expansion of the disturbing function (R) which governs the dynamics of a satellite (natural or artificial) in the Neptune-Triton system. What makes this problem quite unusual, is the fact that a small inner satellite can be strongly disturbed by Triton which is moving in a highly inclined and retrograde orbit. These features are unique in our solar system. Although a lot of retrograde satellites are currently known, all of them have negligible mass and the), do not offer almost any perturbation on the others satellites. However, in the case of the inner satellites of Neptune, Triton is an interesting exception. In a highly inclined orbit, the perturbation it exerts on the neighbouring satellites of Neptune cannot be ignored even for the present scenario. However, in the future, this perturbation will be much more important because due to the tides, the orbit of Triton is contracting, whereas the semi major axes of the remaining inner satellites of Neptune will remain almost unaffected by the tides. In this work we first obtain the disturbing function in the retrograde case. After that, we generalize R for arbitrary inclination. Several numerical tests are presented and a possible future case of resonant configuration is briefly discussed as well. (c) 2005 COSPAR. Published by Elsevier Ltd. All rights reserved.

Disturbances in beta-cell function in impaired fasting glycemia

In a cross-sectional study, we assessed beta-cell function and insulin sensitivity index (ISI) with hyperglycemic clamps (10 mmol/l) in 24 subjects with impaired fasting glycemia (IFG, fasting plasma glucose [FPG] between 6.1 and 7.0 mmol/l), 15 type 2 diabetic subjects (FPG >7.0 mmol/l), and 280 subjects with normal fasting glycemia (NFG, FPG <6.1 mmol/l). First-phase insulin release (0-10 min) was lower in IFG (geometric mean 541 pmol/l (.) 10 min; 95% confidence interval [CI] 416-702 pmol/l (.) 10 min) and in type 2 diabetes (geometric mean 376 pmol/l (.) 10 min; 95% CI 247-572 pmol/l (.) 10 min) than NFG (geometric mean 814 pmol/l (.) 10 min; 95% CI 759-873 pmol/l (.) 10 min) (P < 0.001). Second-phase insulin secretion (140-180 min) was also lower in IFG (geometric mean 251 pmol/l; 95% CI 198-318 pmol/l; P = 0.026) and type 2 diabetes (geometric mean 157 pmol/l; 95% CI 105-235 pmol/l; P < 0.001) than NFG (geometric mean 295 pmol/l; 95% CI 276-315 pmol/l): IFG and type 2 diabetic subjects had a lower ISI (0.15 +/- 0.02 and 0.16 +/- 0.02 mumol/kg fat-free mass [FFM]/min/ pmol/l, respectively) than NFG (0.24 +/- 0.01 mumol/kg FFM/min/pmol/l, P < 0.05). We found a stepwise decline in first-phase (and second-phase) secretion in NFG subjects with progressive decline in oral glucose tolerance (P < 0.05). IFG subjects with impaired glucose tolerance (IGT) had lower first-phase secretion than NFG subjects with IGT (P < 0.02), with comparable second-phase secretion and ISI. NFG and IFG subjects with a diabetic glucose tolerance (2-h glucose >11.1 mmol/l) had a lower ISI than their respective IGT counterparts (P < 0.05). We conclude that the early stages of glucose intolerance are associated with disturbances in beta-cell function, while insulin resistance is seen more markedly in later stages.

Tracing heme in a living cell: hemoglobin degradation and heme traffic in digest cells of the cattle tick Boophilus microplus

Heme is present in all cells, acting as a cofactor in essential metabolic pathways such as respiration and photosynthesis. Moreover, both heme and its degradation products, CO, iron and biliverdin, have been ascribed important signaling roles. However, limited knowledge is available on the intracellular pathways involved in the flux of heme between different cell compartments. The cattle tick Boophilus microplus ingests 100 times its own mass in blood. The digest cells of the midgut endocytose blood components and huge amounts of heme are released during hemoglobin digestion. Most of this heme is detoxified by accumulation into a specialized organelle, the hemosome.We followed the fate of hemoglobin and albumin in primary cultures of digest cells by incubation with hemoglobin and albumin labeled with rhodamine. Uptake of hemoglobin by digest cells was inhibited by unlabeled globin, suggesting the presence of receptor-mediated endocytosis. After endocytosis, hemoglobin was observed inside large digestive vesicles. Albumin was exclusively associated with a population of small acidic vesicles, and an excess of unlabeled albumin did not inhibit its uptake. The intracellular pathway of the heme moiety of hemoglobin was specifically monitored using Palladium-mesoporphyrin IX (Pd-mP) as a fluorescent heme analog. When pulse and chase experiments were performed using digest cells incubated with Pd-mP bound to globin (Pd-mP-globin), strong yellow fluorescence was found in large digestive vesicles 4 h after the pulse. By 8 h, the emission of Pd-mP was red-shifted and more evident in the cytoplasm, and at 12 h most of the fluorescence was concentrated inside the hemosomes and had turned green. After 48 h, the Pd-mP signal was exclusively found in hemosomes. In methanol, Pd-mP showed maximal emission at 550 nm, exhibiting a red-shift to 665 nm when bound to proteins in vitro.The red emission in the cytosol and at the boundary of hemosomes suggests the presence of heme-binding proteins, probably involved in transport of heme to the hemosome. The existence of an intracellular heme shuttle from the digestive vesicle to the hemosome acting as a detoxification mechanism should be regarded as a major adaptation of ticks to a blood-feeding way of life. To our knowledge, this is the first direct observation of intracellular transport of heme in a living eukaryotic cell. A similar approach, using Pd-mP fluorescence, could be applied to study heme intracellular metabolism in other cell types.

Calcifications in a clear cell mucoepidermoid carcinoma: a case report with histological and immunohistochemical findings

The aim of this study was to report an unusual case of mucoepidermoid carcinoma (MEC) in a 39-year-old woman. The tumor showed a prominent population of clear and intermediate basal cells. Clear cells rarely predominate over other cell types. Such cases are called clear cell variant of MEC. The case also revealed a variable amount of calcified material in the tumor mass. Calcifications are rare in clear cell MEC. These structures were periodic acid- Schiff positive and diastase resistant, excluding glycogen origin. Immunohistochemistry was performed, and the epidermoid component was positive for cytokeratin (CK) 7, CK13, CK14, and CK19. The mucous and clear cells presented mild staining for CK7. Cytokeratins 7, 13, and 19 stained luminal cells, and intermediate cells exhibited positivity for CK7, CK14, and vimentin. The origin of the calcifications is speculated to be the result of dystrophic calcification of the amorphous eosinophilic material secreted by intermediate basal cells.

Cell detection at far ranges: supporting T(b) - Z relationships

The Bauru radar detects inner cores of deep convective storm cells at long range, but relevant cell areas are left undetected. A procedure is in development to generate a more complete three-dimensional representation of the cell structure for cost-beneficial hydrological use at larger distances. A set of satellite microwave brightness temperature (T(b)) - radar reflectivity (Z) relationships, basic to the retrieval procedure, has been derived. Copyright (C) 2005 Royal Meteorological Society

Development of Intra- and Interxylary Secondary Phloem in Coccinia indica (Cucurbitaceae)

Stern anatomy and the development of intraxylary phloem were investigated in six to eight years old Coccinia indica L. (Cucurbitaceae). Secondary growth in the stems was achieved by the normal cambial activity. In the innermost part of the thicker stems, xylem parenchyma and pith cells dedifferentiated into meristematic cells at several points. In some of the wider rays, ray cells dedifferentiate and produce secondary xylem and phloem with different orientations and sometimes a complete bicollateral vascular bundle. The inner cambial segments of the bicollateral vascular bundle (of primary growth) maintained radial arrangement even in the mature stems but in most places the cambia were either inactive or showed very few cell divisions. Concomitant with the obliteration and collapse of inner phloem (of bicollateral vascular bundles), parenchyma cells encircling the phloem became meristematic forming a circular sheath of internal cambia. These internal cambia produce only intraxylary secondary phloem centripetally and do not produce any secondary xylem. In the stem, secondary xylem consisted mainly of axial parenchyma, small strands of thick-walled xylem derivatives, i.e. vessel elements and fibres embedded in parenchymatous ground mass, wide and tall rays along with exceptionally wide vessels characteristic of lianas. In thick stems, the axial parenchyma de-differentiated into meristem, which later re-differentiated into interxylary phloem. Fibre dimorphism and pseudo-vestured pits in the vessels are also reported.

Juxtaglomerular cell tumor as a rare cause of hypertension in adults

The juxtaglomerular cell tumor is a cause of secondary hypertension in adults. A 35-year-old female patient suffering from hypertension and low serum potassium had a 3 × 3 cm solid mass at the lower pole of left kidney diagnosed by abdominal sonography. Partial nephrectomy was performed and the postoperatory was uneventful. Normalization of blood pressure was observed within the first month.

Thermodynamic analysis of direct steam reforming of ethanol in molten carbonate fuel cell

Fuel cell as MCFC (molten carbonate fuel cell) operate at high temperatures, and due to this issue, cogeneration processes may be performed, sending heat for own process or other purposes as steam generation in an industry. The use of ethanol for this purpose is one of the best options because this is a renewable and less environmentally offensive fuel, and cheaper than oil-derived hydrocarbons (in the case of Brazil). In the same country, because of technical, environmental and economic advantages, the use of ethanol by steam reforming process have been the most investigated process. The objective of this study is to show a thermodynamic analysis of steam reforming of ethanol, to determine the best thermodynamic conditions where are produced the highest volumes of products, making possible a higher production of energy, that is, a most-efficient use of resources. To attain this objective, mass and energy balances are performed. Equilibrium constants and advance degrees are calculated to get the best thermodynamic conditions to attain higher reforming efficiency and, hence, higher electric efficiency, using the Nernst equation. The advance degree of reforming increases when the operation temperature also increases and when the operation pressure decreases. But at atmospheric pressure (1 atm), the advance degree tends to the stability in temperatures above 700°C, that is, the volume of supplemental production of reforming products is very small for the high use of energy resources necessary. Reactants and products of the steam-reforming of ethanol that weren't used may be used for the reforming. The use of non-used ethanol is also suggested for heating of reactants before reforming. The results show the behavior of MCFC. The current density, at same tension, is higher at 700°C than other studied temperatures as 600 and 650°C. This fact occurs due to smaller use of hydrogen at lower temperatures that varies between 46.8 and 58.9% in temperatures between 600 and 700°C. The higher calculated current density is 280 mA/cm 2. The power density increases when the volume of ethanol to be used also increases due to higher production of hydrogen. The highest produced power at 190 mW/cm 2 is 99.8, 109.8 and 113.7 mW/cm2 for 873, 923 and 973K, respectively. The thermodynamic efficiency has the objective to show the connection among operational conditions and energetic factors, which are some parameters that describes a process of internal steam reforming of ethanol.

Fine needle aspiration biopsy of gastrointestinal stromal tumor presenting as an umbilical mass (Sister Mary Joseph's nodule)

The Sister Mary Joseph (SMJ) nodule is a clinical sign of metastatic cancer involving the umbilicus. The vast majority of these instances represent adenocarcinomas arising from ovarian or colorectal primaries. We present a patient who presented with ascites and the SMJ lesion that turned out to be a metastatic gastrointestinal stromal tumor after fine needle aspiration biopsy was performed. The lesion was subsequently histologically confirmed. Gastrointestinal stroma tumor involving the umbilicus is exceedingly uncommon and only rarely presents in this fashion. The cytomorphological features, differential diagnosis, and comparison with the tissue specimen are made. © 2006 Elsevier Inc. All rights reserved.

Renal cell carcinoma with cutaneous metastasis in a dog

The present report describes the occurrence of skin metastasis in a dog with primary renal carcinoma. The clinical findings included progressive weight loss, inappetence, palpable abdominal mass and cutaneous nodules. The histopathological examination was carried out after unilateral nephrectomy and was confirmed by immunohistochemistry. The immunohistochemical and histopathological findings characterized this case as renal cell carcinoma with skin metastasis.

Visceral mast cell tumor and mastocythemia in a dog

Mast cell tumor manifests as a localized proliferation of mast cells in the skin, or less frequently as a systemic disorder, which may be accompanied by the presence of neoplastic mast cells in the peripheral blood (mastocythemia). In some cases, the neoplastic circulating mast cells originate in the bone marrow, designated as mast cell leukemia, rarely observed in dogs, or the cells may arise from visceral mast cell tumors, characterizing systemic mastocytosis. The aim of this report was to describe a case of a six-year-old female German shepherd dog presenting with history of anorexia, hematemesis and diarrhea. The blood work revealed intense mastocythemia (43%), with degranulated mast cells, and anisocytosis. At necropsy, white nodular lesions in the thymic region and an infiltrative mass in mesenteric and abdominal lymph nodes were observed. Those lymph nodes were enlarged and off-white. Histopathological examination revealed neoplastic mast cells in the liver, spleen, lymph nodes, kidneys, lungs, gastric and enteric mucosae, and adrenal glands. The clinical, hematological and histopathological findings were compatible with mastocythemia, associated with a moderately differentiated visceral mast cell tumor.

Optimal single-embryo mass spectrometry fingerprinting

In pre-implantation embryos, lipids play key roles in determining viability, cryopreservation and implantation properties, but often their analysis is analytically challenging because of the few picograms of analytes present in each of them. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) allows obtaining individual phospholipid profiles of these microscopic organisms. This technique is sensitive enough to enable analysis of individual intact embryos and monitoring the changes in membrane lipid composition in the early stages of development serving as screening method for studies of biology and biotechnologies of reproduction. This article introduces an improved, more comprehensive MALDI-MS lipid fingerprinting approach that considerably increases the lipid information obtained from a single embryo. Using bovine embryos as a biological model, we have also tested optimal sample storage and handling conditions before the MALDI-MS analysis. Improved information at the molecular level is provided by the use of a binary matrix that enables phosphatidylcholines, sphingomyelins, phosphatidylserines, phosphatidylinositols and phosphoethanolamines to be detected via MALDI(±)-MS in both the positive and negative ion modes. An optimal MALDI-MS protocol for lipidomic monitoring of a single intact embryo is therefore reported with potential applications in human and animal reproduction, cell development and stem cell research. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.