<Emphasis Type="Italic">Ctenomys lami</Emphasis>: The highest chromosome variability in<Emphasis Type="Italic">Ctenomys</Emphasis> (Rodentia,Ctenomyidae) due to a centric fusion/fission and pericentric inversion system

Ctenomys lami Freitas, 2001 is an endemic species of rodent inhabiting the Coastal Plain of southern Brazil, along a narrow line of old dunes formed in the Pleistocene. This species has five different diploid numbers (2n=54, 55a, 55b, 56a, 56b, 57 and 58) and ten different autosomal fundamental numbers (FNa=74, 75, 76, 77, 78, 79, 80, 81, 82, and 84). In a sample of 102 specimens, the combined 2n and FNa formed 26 different karyotypes. The diploid number variation was due to Robertsonian rearrangements that occur in pairs 1 and 2, and the variation of NFas was due to pericentric inversions. The distribution of diploid number variation along the 78 km line of collection sites reveals four population blocks: block A with 2n=54, 55a, and 56a; block B with 2n=57 and 58; block C with 2n=54 and 55a; and block D with 2n=56b and 55b. The inversion system lacks geographic structure with a random distribution of inversions along the population blocks. A very narrow hybrid zone is hypothesized between blocks A and B. Blocks B and C are separated by a geographic barrier, and another hybrid zone is found between blocks C and D. My findings suggest that this species is undergoing a process of speciation due to geographic isolation.     

Halothane Increases Non-vesicular [<Superscript>3</Superscript>H]dopamine Release from Brain Cortical Slices

Experimental data suggest that halothane anesthesia is associated with significant changes in dopamine (DA) concentration in some brain regions but the mechanism of this effect is not well known. Rat brain cortical slices were labeled with [³H]DA to further characterize the effects of halothane on the release of this neurotransmitter from the central nervous system. Halothane induced an increase on the release of [³H]DA that was dependent on incubation time and anesthetic concentration (0.012, 0.024, 0.048, 0.072 and 0.096 mM). This effect was independent of extracellular or intracellular calcium. In addition, [³H]DA release evoked by halothane was not affected by TTX (blocker of voltage-dependent Na⁺ channels) or reserpine (a blocker of vesicular monoamine transporter). These data suggest that [³H]DA release induced by halothane is non-vesicular and would be mediated by the dopamine transporter (DAT) and norepinephrine transporter (NET). GBR 12909 and nomifensine, inhibitors of DAT, decreased the release of [³H]DA evoked by halothane. Nisoxetine, a blocker of NET, reduced the release of [³H]DA induced by halothane. In addition, GBR 12909, nisoxetine and, halothane decrease the uptake of [³H]DA into rat brain cortical slices. A decrease on halothane-induced release of [³H]DA was also observed when the brain cortical slices were incubated at low temperature and low extracellular sodium, which are known to interfere with the carrier-mediated release of the neurotransmitter. Ouabain, a Na⁺/K⁺ ATPase pump inhibitor, which induces DA release through reverse transport, decreased [³H]DA release induced by halothane. It is suggested that halothane increases [³H]DA release in brain cortical slices that is mediated by DAT and NET present in the plasma membrane.     

Irradiated cationic mesoporphyrin induces larger damage to isolated rat liver mitochondria than the anionic form

The action of irradiated cationic Fe(III)TMPyP and anionic Fe(III)TPPS4 forms of mesoporphyrins on mitochondrial functions was investigated using experimental conditions that caused minimal effects on mitochondria in the dark. Treatment of mitochondria with 1 microM Fe(III)TMPyP for 2 min decreased the respiratory control by 3% in the dark and 28% after irradiation. Fe(III)TPPS4 (1 microM) had no significant effect on respiratory control under any of the above conditions. Both porphyrins increased the mitochondrial production of reactive oxygen species in the presence of Ca2+; however, the effect of Fe(III)TMPyP was significantly stronger. In both cases, this overproduction was associated with membrane lipid peroxidation. It was also observed that the association constant of Fe(III)TMPyP with mitochondria was 11 times higher than that of Fe(III)TPPS4. In conclusion, the damage to isolated mitochondria induced by Fe(III)TMPyP under illumination was larger than by Fe(III)TPPS4, probably because its cationic charge favors association with the mitochondrial membrane. This is supported by the decrease in the association constant of Fe(III)TMPyP with mitochondria in higher salt medium.     

CXC chemokine receptor 4 is expressed in uveal malignant melanoma and correlates with the epithelioid-mixed cell type

PURPOSE: Although relatively rare, uveal melanoma is the most common ocular tumor of adults. Up to half of uveal melanoma patients die of metastatic disease. CXCR4, a chemokine receptor, is a prognostic factor in cutaneous melanoma involved in angiogenesis and metastasis formation. The aim of this study was to evaluate the expression of CXCR4 in uveal melanoma. METHODS: CXCR4 was detected by immunohistochemistry in 44 samples of uveal melanoma. Staining was categorized into three semiquantitative classes based on the rate of stained (positive) tumor cells: absence of staining, <50% of cell (+) and >50% (++). Correlations between CXCR4 expression, data on patient and tumor features were studied by contingency tables and the chi2 test. Time-to-event curves were studied using the Kaplan-Meier method. Univariate analysis was performed using the log-rank test. Ninety-five percent confidence intervals (95% CI) of hazard ratios were also reported. RESULTS: Staining for CXCR4 protein was absent in 18 tumors (40.9%), present in <50% of cells in 19 (43.2%) and in >50% of cells in 7 (15.9%) tumors. CXCR4 expression correlated to the epithelioid-mixed cell type (P=0.030). No statistically significant relation emerged between CXCR4 expression, largest tumor diameter (LTD) and extracellular matrix patterns as evaluated through histological patterns stained with periodic acid-Schiff (PAS). Events occurred in 2 out of 18 patients (11.1%) with negative tumors (2 deaths), in 3 out of 19 patients (15.8%) with <50% of positive tumor cells (2 deaths and 1 occurrence of metastases) and in 1 out of 7 patients (14.3%) with >50% of positive tumor cells (1 occurrence of metastases). The cell type (P=0.0457) but not CXCR4 showed prognostic value at univariate analysis. CONCLUSION: This study shows that CXCR4 is commonly expressed in uveal melanoma and correlates with cell type a well-established prognostic factor.     

Spatial distribution and habitat of the Anavilhanas Archipelago bird community in the Brazilian Amazon

This study is the first to present a quantitative survey of bird species occurring in the archipelago of Anavilhanas, located in the Rio Negro in the Brazilian Amazon and is part of the Anavilhanas Ecological Station. We asked whether bird community composition is similar among the islands, and between islands and areas dominated by the surrounding upland terra firme forest on the left (east) margin of the Rio Negro. The surveys were conducted in November and December of 1988, using two complementary methods with mist nets and boat transects. A total of 232 bird species was found for Anavilhanas including a survey done in 1998. The families Tyrannidae and Thamnophilidae showed the highest number of species (16.4% and 9.0% of the total respectively). Some species not well known or having limited distributions are relatively frequently encountered in the archipelago, such as Spizastur melanoleucus, Mitu tomentosa, Phaethornis rupurumii, Xiphorhynchus kienerii, Thamnophilus nigrocinereus, Myrmotherula klagesi, Myrmoborus lugubris, Pipra filicauda, and Cephalopterus ornatus. Hybrid Multidimensional Scaling (HMDS) ordination analysis indicated that the bird community composition is similar among islands. However, the bird community composition on the islands was significantly different from that in sites of terra firme forest at Rio Negro margins. Anavilhanas is a unique ecological system in the Amazon and has it own avifauna.     

CD16 promotes Escherichia coli sepsis through an FcR gamma inhibitory pathway that prevents phagocytosis and facilitates inflammation

Sepsis, a leading cause of death worldwide, involves proinflammatory responses and inefficient bacterial clearance. Phagocytic cells play a crucial part in the prevention of sepsis by clearing bacteria through host innate receptors. Here we show that the FcRgamma adaptor, an immunoreceptor tyrosine-based activation motif (ITAM)-bearing signal transduction subunit of the Fc receptor family, has a deleterious effect on sepsis. FcRgamma(-/-) mice show increased survival during peritonitis, owing to markedly increased E. coli phagocytosis and killing and to lower production of the proinflammatory cytokine tumor necrosis factor (TNF)-alpha. The FcRgamma-associated receptor that inhibits E. coli phagocytosis is FcgammaRIII (also called CD16), and its absence protects mice from sepsis. FcgammaRIII binds E. coli, and this interaction induces FcRgamma phosphorylation, recruitment of the tyrosine phosphatase SHP-1 and phosphatidylinositide-3 kinase (PI3K) dephosphorylation. Decreased PI3K activity inhibits E. coli phagocytosis and increases TNF-alpha production through Toll-like receptor 4. We identified the phagocytic receptor negatively regulated by FcRgamma on macrophages as the class A scavenger receptor MARCO. E. coli-FcgammaRIII interaction induces the recruitment of SHP-1 to MARCO, thereby inhibiting E. coli phagocytosis. Thus, by binding FcgammaRIII, E. coli triggers an inhibitory FcRgamma pathway that both impairs MARCO-mediated bacterial clearance and activates TNF-alpha secretion.     

Blood and inflammatory cells of the lungfish Lepidosiren paradoxa

A special interest exists concerning lungfish because they may possess characteristics of the common ancestor of land vertebrates. However, little is known about their blood and inflammatory cells; thus the fine structure, cytochemistry and differential cell counts of coelomic exudate and blood leucocytes were studied in Lepidosiren paradoxa. Blood smear analyses revealed erythrocytes, lymphocytes, monocytes, polymorphonuclear agranulocytes, thrombocytes and three different granulocytes. Blood monocytes and lymphocytes had typical vertebrate morphology. Thrombocytes had large vacuoles filled with a myelin rich structure. The polymorphonuclear agranulocyte had a nucleus morphologically similar to the human neutrophil with no apparent granules. Types I and II granulocytes had eosinophilic granules. Type I granulocytes had round or elongated granules heterogeneous in size, while type II had granules with an electron dense core. Type III granulocyte had many basophilic granules. The order of frequency was: type I granulocyte, followed by lymphocyte, type II granulocyte, monocyte, polymorphonuclear agranulocyte and type III granulocyte. Peroxidase localized mainly at the periphery of the granules from type II granulocytes, while no peroxidase expression was detected in type I granulocytes. Alkaline phosphatase was localized in the granules of type II granulocyte and acid phosphatase cytochemistry also labelled a few vacuoles of polymorphonuclear agranulocyte. About 85% of the coelomic inflammatory exudate cell population was type II granulocyte, 10% polymorphonuclear agranulocyte and 5% macrophages as judged by the nucleus and granule morphology. These results indicate that this lungfish utilises type II granulocytes as its main inflammatory granulocytes and that the polymorphonuclear agranulocyte may also be involved in the inflammatory response. The other two granulocytes appear similar to the mammalian eosinophil and basophil. In summary, this lungfish appears to possess the typical inflammatory granulocytes of teleosts, however, further functional studies are necessary to better understand the polymorphonuclear agranulocyte.     

Intermittent exercise as a conditioning activity to induce postactivation potentiation

Postactivation potentiation (PAP) is defined as a short-term increase in voluntary muscle activation following a previous conditioning activity (CA). Controversy about PAP is mostly attributed to the characteristics of the CA and the training status of the subjects. While some studies have found that PAP can be induced by series of 5-10 second maximal voluntary isometric contractions or near maximal dynamic contractions (e.g., 3-5 repetition maximum), others have failed to do so. On the other hand, some studies suggest that intermittent contractions can also induce PAP. However, even though PAP was observed, its duration was not taken into account, leaving ground for further investigations. The purpose of this study was threefold: (a) to verify if PAP can progressively enhance performance of voluntary actions throughout a set of intermittent contractions; (b) to verify PAP duration when induced by an intermittent contractions protocol; and (c) to verify if PAP effects were reproducible in different sessions when induced by intermittent contractions. Ten physically active men, not engaged in strength training, underwent 5 randomized experimental sessions, during which they performed a set of 10 unilateral knee extensions (KE) (1 every 30 seconds) at 60 degrees x s(-1) in an isokinetic dynamometer. Peak torque was evaluated over the 10 unilateral KE and at the randomized intervals of 4, 6, 8, 10, and 12 minutes post CA. Peak torque was potentiated 1.3 (+/-0.79) N x m per unilateral KE, and the potentiation effect persisted for 12 minutes after the last contraction. These findings were reproduced in all 5 experimental sessions. Thus, intermittent conditioning activities seem to be an effective way to produce PAP. However, these activities should be tested in a more real world situation to verify the applicability as a warm-up routine.     

Molecular evolution of PIII-SVMP and RGD disintegrin genes from the genus Crotalus

Several types of disintegrins have been isolated from Crotalus spp rattlesnakes, including RGD disintegrins, and PIII-SVMPs. We isolated six cDNAs from snake venom glands using RT-PCR. Three RGD disintegrins (atroxatin, mojastin, and viridistatin) and three PIII-SVMPs (catroriarin, scutiarin, and viristiarin) cDNAs were isolated from the rattlesnakes Crotalus atrox, Crotalus scutulatus scutulatus, and Crotalus viridis viridis, respectively. Atroxatin and Viridistatin shared 90% amino acid identity to each other, and 87% identity to Mojastin. Scutiarin and Viristiarin were identical. All PIII-SVMPs isolated in this study shared the highest amino acid identity with Catrocollastatin. cDNA and protein sequences for RGD disintegrins, one MVD disintegrin, and PIII-SVMPs of the genus Crotalus (present in the NCBI database), were used in phylogenetic analysis. Neighbor-joining analysis of PIII-SVMP and RGD/MVD disintegrin-coding DNA sequences showed that these groups of genes separate into separate clades. A Phi(ST) pairwise comparison and Analysis of Molecular Variance (AMOVA) between PIII-SVMPs and RGD/MVD disintegrins showed significant genetic differences. Mutations observed in ten of the cDNAs analyzed did not affect Cys-coding sequences. Our K(A)/K(S) data suggest that rapid evolution occurred between the genes coding for PIII-SVMPs resulting, in the production of RGD disintegrin-coding genes. However, once these genes diverged, mutations in the PIII-SVMP-coding genes were accumulated less frequently.