Population structure of Uncancylus concentricus (d'Orbigny, 1835) (Ancylidae, Pulmonata, Basommatophora) in the Multiple Use Reserve Martín García Island, Upper Río de la Plata, Argentina.

The present work analyzes the population dynamics of Uncancylus concentricus in natural conditions in the northeastern coastal area of the Multiple Use Natural Reserve Isla Martín García (Beach of Basural), Buenos Aires, Argentina. Martín García Island is located in the Upper Río de La Plata, to the south of the mouth of the Uruguay River (34° 11' 25" S and 58° 15'38" W). Monthly collections were made from August 2005 to December 2006. The size frequency of the U. concentricus population throughout the sampling period ranges from 1.2 to 8.3 m. The changes in the size frequencies throughout the months surveyed indicate a variable distribution pattern because of the increase and decrease in water flow, but we observed a peak in the frequency of individuals within the size range 3.5 to 4.5 mm in the population. It can be concluded that this species lives for at least a year in the wild since the juveniles are highly abundant in spring and summer.

Mollusks (Gastropoda and Bivalvia) of the Multiple-Use Reserve Martín García Island, Río de la Plata River: biodiversity and ecology.

The Island of Martin Garcia is located in the Upper Río de la Plata, to the south of mouth the Uruguay River. The aim of the present study was to analyse the biodiversity of the island freshwater mollusks and their relationships to environmental variables. Twelve sampling sites were selected, five were along the littoral section of the island and seven were Inland ponds. Seven major environmental variables were measured: water and air temperature, percentage of oxygen saturation, dissolved oxygen, electrical conductivity, total dissolved solids and pH. Twenty-seven mollusk species were found, Antillorbis nordestensis, Biomphalaria tenagophila tenagophila , B. t. guaibensis, B. straminea, B. peregrina, Drepanotrema kermatoides, D. cimex, D. depressissimum, Chilina fluminea, C. rushii, C. megastoma, Uncancylus concentricus, Hebetancylus moricandi, Stenophysa marmorata, Heleobia piscium, H. parchappii, Potamolithus agapetus, P. buschii, P. lapidum, Pomacea canaliculata, P. megastoma, Asolene platae, Corbicula fluminea, Eupera platensis, Pisidium sterkianum, P. taraguyense and Limnoperna fortunei. UPGMA clustering of species based on their occurrence in different ecological conditions revealed two main species groups. The Canonical Correspondence Analysis suggests that the species distribution is related to the physico-chemical condition of water. Axis two of the ordination diagram displayed the approximately 95.6% of the correlation between species and environmental variables. Dissolved oxygen, conductivity, water temperature and pH showed the highest fluctuations during the sampling period. The species richness (S) showed relationships mainly with water temperature and conductivity. The biodiversity of the gastropods and bivalves from Martín García Island amounts to up to 26 species. Among the Gastropoda, the Planorbidae family made the most sizeable contribution. The Lithogliphidae P. agapetus (26.28%) and P. buschii (9.50%) showed the highest relative frequencies of occurrence within the littoral environments, while the Planorbidae D. cimex (23.83%) and D. kermatoides (11.59%) likewise did so in the inland ponds.

2-deoxyglucose-induced toxicity is regulated by Bcl-2 family members and is enhanced by antagonizing Bcl-2 in lymphoma cell lines.

Targeting altered cancer cell metabolism with the glycolysis inhibitor, 2-deoxyglucose (2DG), is a viable therapeutic strategy, but the effects of 2DG on lymphoma cells and the mechanism of action are unknown. Five T-cell lymphoma lines and two B-cell lymphoma lines were shown to be highly sensitive to 2DG. Examination of the cell death pathway demonstrated pro-apoptotic protein Bax 'activation' and caspase cleavage in 2DG-treated cells. However, Q-VD-OPh, a potent inhibitor of caspase activity provided minimal protection from death. In contrast, overexpressing the anti-apoptotic protein Bcl-2 dramatically enhanced the survival of 2DG-treated cells that was negated by a Bcl-2 antagonist. BH3-only members, Bim and Bmf, were upregulated by 2DG, and shRNAs targeting Bim protected from 2DG toxicity demonstrating that Bim is a critical mediator of 2DG toxicity. 2DG also induced GADD153/CHOP expression, a marker of endoplasmic reticulum (ER) stress and a known activator of Bim. Mannose, a reagent known to alleviate ER stress, transiently protected from 2DG-induced cell death. Examination of the effects of 2DG on energy metabolism showed a drop in ATP levels by 30 min that was not affected by either Bcl-2 or mannose. These results demonstrate that ER stress appears to be rate limiting in 2DG-induced cell death in lymphoma cells, and this cell killing is regulated by the Bcl-2 family of proteins. Bcl-2 inhibition combined with 2DG may be an effective therapeutic strategy for lymphoma.

Biodiversity and ecology of Hirudinea (Annelida) from the Natural Reserve of Isla Martín García, Río de la Plata, Argentina.

The Island of Martin Garcia lies at the confluence of the Uruguay and Paraná Rivers (upper Río de la Plata). This island is an outcrop of the crystalline basement. Due to basalt exploitation the island exhibits several ponds covered by carpets of free-floating macrophytes. Seven major environmental variables were measured: water and air temperature, percentage of oxygen saturation, dissolved oxygen, electrical conductivity, total dissolved solids and pH. Eleven leech species were found, Helobdella striata, H. diploides, H. adiastola and H. hyalina were new records. UPGMA clustering of species based on their occurrence in different ecological conditions revealed three main species groups. Canonical Correspondence Analysis (CCA) explained 97.2% of the correlation between species and environmental variables. H. triserialis shows the widest range of tolerance, H. hyalina shows positive relationships to conductivity; H. diploides shows a high correlation with dissolved oxygen, H. striata, H. lineata, and S. similis are negatively correlated with water temperature, and H. simplex is positively correlated with pH. Relationships between the species richness (S) and the sampling sites were negatively correlated with water temperature and positively correlated with dissolved oxygen. Leech biodiversity from the water bodies of Martín García Island, shows a great diversity of species and a wide plasticity regarding the characteristics of the environmental factors considered.

Distribution of Deroceras reticulatum (Müller, 1774) (Pulmonata Stylommatophora) in Argentina with first record of the Reserva de Usos Múltiples Isla Martin Garcia, Río de la Plata superior.

Deroceras reticulatum is a misanthropic European species spread widely throughout South America. At the moment this species is considered a 'pest' in direct sowing such as maize, soybean, sunflower, wheat, alfalfa and clovers, among others. The aim of this paper is to report the first record of D. reticulatum in the Reserva de Usos Multiples Isla Martín García, Buenos Aires province and to provide information about this species distribution in five Argentina provinces.

Individual growth of Heleobia piscium in natural populations (Gastropoda: Cochliopidae) from the multiple use natural Reserve Isla Martin Garcia, Buenos Aires, Argentina.

The present work analyses the individual growth of Heleobia piscium in natural conditions in coastal drainage channels of the Multiple Use Natural Reserve Isla Martín García, Buenos Aires, Argentina. Isla Martín García is located in the Upper Río de la Plata, to the south of the mouth of the Uruguay river (34 degrees 11' 25" S and 58 degrees 15' 38" W). Monthly collections were made from July 2005 to July 2006 in the eastern part of the island (Arena Beach). The population of H. piscium showed a complex and dynamic structure of sizes during a long period of the annual cycle. Two cohorts could be detected. The Bertalanffy growth equation was: Lt = 6 (1-e -1.85 (t+0.38)) and Lt = 3.9 (1-e -0.19 (t+4.84)) for cohorts 1 and 2, respectively. The pattern of population growth displayed a staggered model, where the greatest growth is observed during the summer. The reproductive period occurred during six months, from the beginning of summer to middle of fall. Based on only one reproductive effort, this pattern is not similar to that of other cogeneric species already studied.

Individual growth of Heleobia piscium in natural populations (Gastropoda: Cochliopidae) from the multiple use natural Reserve Isla Martin Garcia, Buenos Aires, Argentina / Crescimento individual de Heleobia piscium em populações naturais (Gastropoda: Cochliopidae) da Reserva Natural de usos múltiplos Ilha Martín García, Buenos Aires, Argentina

The present work analyses the individual growth of Heleobia piscium in natural conditions in coastal drainage channels of the Multiple Use Natural Reserve Isla Martín García, Buenos Aires, Argentina. Isla Martín García is located in the Upper Río de la Plata, to the south of the mouth of the Uruguay river (34° 11' 25" S and 58° 15' 38" W). Monthly collections were made from July 2005 to July 2006 in the eastern part of the island (Arena Beach). The population of H. piscium showed a complex and dynamic structure of sizes during a long period of the annual cycle. Two cohorts could be detected. The Bertalanffy growth equation was: Lt = 6 (1-e -1.85 (t+0.38)) and Lt = 3.9 (1-e -0.19 (t+4.84)) for cohorts 1 and 2, respectively. The pattern of population growth displayed a staggered model, where the greatest growth is observed during the summer. The reproductive period occurred during six months, from the beginning of summer to middle of fall. Based on only one reproductive effort, this pattern is not similar to that of other cogeneric species already studied.

O presente trabalho analisa o crescimento individual de Heleobia piscium em condições naturais em poças costeiras da Reserva Natural de Usos Múltiplos Ilha Martín García, Buenos Aires, Argentina. A Ilha Martín García está localizada no Rio da Prata superior, ao sul da desembocadura do rio Uruguai (34° 11' 25" S e 58°15' 38" W). Amostras mensais foram analisadas entre os meses de julho de 2005 e julho de 2006, no setor este da ilha (Praia de Arena). A população de Heleobia piscium se caracterizou por uma complexa e dinâmica estrutura de talhas ao longo de grande parte do ciclo anual. Duas cohortes puderam ser detectadas. A equação de von Bertalanffy para a cohorte 1 foi: Lt = 6 (1-e -1.85 (t+0.38)). Para a cohorte 2: Lt = 3.9 (1-e -0.19 (t+4.84)). O padrão de desenvolvimento da população mostra um modelo escalonado, estendendo-se o período de maior crescimento durante toda a estação do verão. A temporada reprodutiva se manifestou durante seis meses, desde o princípio do verão até meados do outono. Este padrão baseado em um único esforço reprodutivo não se assemelha ao de outras espécies congenéricas já estudadas.

Radular ultrastructure of South American Ampullariidae (Gastropoda: Prosobranchia)

The radula of five species of South American Ampullariidae was analysed by Scanning Electron Microscope (SEM) with the purpose of enlarging new studies on the systematic of this family. The studied species were Pomacea -canaliculata (Lamarck, 1822), Pomacea scalaris (d'Orbigny, 1835), Pomella (P.) megastoma (Gray, 1847), Asolene (A.) platae (Maton, 1809) and Felipponea neritiniformis (Dall, 1919). The central tooth shows different attributes which provide the means for generic determination; the analysis of the central tooth, the lateral and marginal ones by SEM adds further information for species differentiation.

A rádula de cinco espécies sul-americanas de Ampullariidae é analisada ao microscópio eletrônico de varredura com objetivo de fornecer novos subsídios à sistemática desta família. As espécies estudadas são Pomacea canaliculata (Lamarck, 1822), Pomacea scalaris (d'Orbigny, 1835), Pomella (P.) megastoma (Gray, 1847), Asolene (A.) platae (Maton, 1809) e Felipponea neritiniformis (Dall, 1919). O dente central apresenta novos subsídios à determinação genérica, o estudo ao microscópio eletrônico os dentes central, laterais e marginais acrescentam novos subsídios na determinação das espécies.

Radular ultrastructure of South American Ampullariidae (Gastropoda: Prosobranchia).

The radula of five species of South American Ampullariidae was analysed by Scanning Electron Microscope (SEM) with the purpose of enlarging new studies on the systematic of this family. The studied species were Pomacea canaliculata (Lamarck, 1822), Pomacea scalaris (d'Orbigny, 1835), Pomella (P.) megastoma (Gray, 1847), Asolene (A.) platae (Maton, 1809) and Felipponea neritiniformis (Dall, 1919). The central tooth shows different attributes which provide the means for generic determination; the analysis of the central tooth, the lateral and marginal ones by SEM adds further information for species differentiation.

Implementing a network for electronic surveillance reporting from public health reference laboratories: an international perspective.

Electronic data reporting from public health laboratories to a central site provides a mechanism for public health officials to rapidly identify problems and take action to prevent further spread of disease. However, implementation of reference laboratory systems is much more complex than simply adopting new technology, especially in international settings. We describe three major areas to be considered by international organizations for successful implementation of electronic reporting systems from public health reference laboratories: benefits of electronic reporting, planning for system implementation (e.g., support, resources, data analysis, country sovereignty), and components of system initiation (e.g., authority, disease definition, feedback, site selection, assessing readiness, problem resolution). Our experience with implementation of electronic public health laboratory data management and reporting systems in the United States and working with international organizations to initiate similar efforts demonstrates that successful reference laboratory reporting can be implemented if surveillance issues and components are planned.

Peptide length variants p2Ca and QL9 present distinct conformations to L(d)-specific T cells.

Recent advances have provided insights into how the TCR interacts with MHC/peptide complexes and a rationale to predict optimal epitopes for MHC binding and T cell recognition. For example, peptides of nine residues are predicted to be optimal for binding to H2-L(d), although 8 mer epitopes have also been identified. It has been predicted that 8 mer and 9 mer length variant peptides bound to L(d) present identical epitopes to T cells. However, in contrast to this prediction, we demonstrate here that the 8 mer peptide p2Ca and its 9 mer length variant QL9, extended by an N-terminal glutamine, assume distinct conformations when bound to L(d). We generated self-L(d)-restricted CTL clones specific for p2Ca that recognize L(d)/QL9 poorly if at all. This result is in sharp contrast to what has been observed with L(d)-alloreactive T cells that possess a much higher affinity for L(d)/QL9 than for L(d)/p2Ca. Alanine substitutions of the N-terminal residues of the QL9 peptide rescue detection by these self-L(d)/p2Ca-specific T cells, but decrease recognition by the L(d)-alloreactive 2C T cell clone. In addition, 2C T cell recognition of the p2Ca peptide is affected by different alanine substitutions compared with 2C T cell recognition of the QL9 peptide. These data clearly demonstrate that the p2Ca and QL9 peptides assume distinct conformations when bound to L(d) and, furthermore, demonstrate that there is flexibility in peptide binding within the MHC class I cleft.

Testosterone delivery using glutamide-based complex high axial ratio microstructures.

Complex high axial ratio microstructures (CHARMs) were evaluated for delivery of testosterone in vivo. Methods to incorporate testosterone included noncovalent mixing and covalent attachment of testosterone to the lipid to form a prodrug monomer. When prepared by covalent attachment, testosterone-loaded CHARMs were resistant to in vitro spontaneous hydrolysis; when injected into rats, testosterone was released with biphasic kinetics consisting of a burst followed by a much slower phase. Some CHARM material associated with testosterone persisted at the site of injection for at least 9 days.

Novel reduced benzo[j]fluranthen-3-ones from Cladosporium cf. cladosporioides with cytokine production and tyrosine kinase inhibitory properties.

A series of reduced benzo[j]fluoranthen-3-ones (1-4) was isolated from fermentations of a fungal strain CBUK20700 (CBS 100220), classified as Cladosporium cf. cladosporioides, during a microbial extract screening programme to identify inhibitors of anti-CD28-induced interleukin-2 (IL-2) production by Jurkat E6-1 cells as potential immunosuppressive agents. These compounds were also found to be tyrosine kinase inhibitors. The structures of compounds 1-4 were elucidated by spectroscopic methods including the HMQC, HMBC and NOESY NMR experiments. The most potent compound in the series, (6bS,7R,8S)-7-methoxy-4,8,9-trihydroxy-1,6b,7,8-tetrahydro-2H-benzo[j]fluoranthen-3-one (1) inhibited anti-CD28-induced IL-2 production and Abl tyrosine kinase with IC50 values of 400 and 60 nM respectively. The 6b-stereoisomeric 2 was a moderate inhibitor of both IL-2 production and Abl tyrosine kinase while the 8-oxo derivative 3 was inactive in both assays. The 8-O-methyl ether 4 was a moderate inhibitor of IL-2 production but exhibited potent inhibition of Abl tyrosine kinase with an IC50 of 45 nM.

Nitric oxide synthase inhibitors decrease coronary sinus-free radical concentration and ameliorate myocardial stunning in an ischemia-reperfusion model.

OBJECTIVES: Our objective was to determine the effect of a nitric oxide synthase inhibitor, NG-nitro-L-arginine (L-NNA) on free radical generation and myocardial contractility after ischemia-reperfusion. BACKGROUND: Cardiotoxic free radicals are generated by ischemia-reperfusion sequences. Nitric oxide reacts with superoxide radical to form peroxynitrite, which generates additional free radicals. Our hypothesis was that by inhibiting NO production, free radical formation will be diminished, which should be cardioprotective. METHODS: We studied 32 dogs. Coronary occlusion-reperfusion (20 min each) sequences were created by intracoronary balloon angioplasty inflation-deflation. Using electron paramagnetic resonance, we monitored the coronary sinus concentration of ascorbate free radical (Asc*-), a measure of total oxidative flux. The L-NNA (4.8 mg/kg total) was infused intravenously during occlusion-reperfusion; control dogs received saline. Immunohistochemical staining demonstrated the peroxynitration product nitrotyrosine. RESULTS: In the control dogs Asc*- rose from 3.2 +/- SD 0.5 nmol/l to 4.8 +/- 1.1 nmol/l with reperfusion, a 50% rise. With L-NNA the Asc*- rose from 3.2 +/- 0.9 nmol/l to 4.0 +/- 1.2 nmol/l, a 25% rise (p < 0.01, L-NNA vs. control). Echocardiographic left ventricular fractional area shortening (FAS) in the control dogs declined from 38 +/- 19% (baseline) to 26 +/- 14% (ischemia), and to 22 +/- 11% with reperfusion (p < 0.01 vs. baseline). With L-NNA, FAS declined from 36 +/- 13% (baseline) to 27 +/- 12% (ischemia) but then rose to 33 +/- 14 with reperfusion (p = NS vs. baseline). Nitrotyrosine was present in the myocardium subjected to ischemia-reperfusion, but almost absent in dogs receiving L-NNA. Myocardial perfusion was not altered by L-NNA. CONCLUSIONS: The NO synthase inhibitors decrease coronary sinus free radical concentration and ameliorate myocardial stunning after ischemia-reperfusion.

The generality of architectural isomerism in designer inclusion frameworks.

We describe herein new structural isomers of a lamellar host system based on organodisulfonate "pillars" that connect opposing hydrogen-bonded sheets, consisting of topologically complementary guanidinium (G) ions and sulfonate (S) groups, to generate inclusion cavities between the sheets. These new isomers-zigzag brick, double brick, V-brick, and crisscross bilayer-expand significantly on our earlier report of architectural isomerism displayed by the discrete bilayer and simple brick forms. We demonstrate here that the discrete bilayer-simple brick isomerism, which was limited to several host-guest combinations based on the G(2)(4,4'-biphenyldisulfonate) host and one pair of compounds based on the G(2)(2,6-naphthalenedisulfonate), can be generalized to other organodisulfonate pillars. Furthermore, in many cases the selectivity toward the different framework isomers reflects a rather systematic templating role of the guest molecules and host-guest recognition during assembly of the lattice. We also describe a convenient approach to identifying and classifying the innumerable possible host architectures based upon the pillar projection topologies for the GS sheets and the intersheet connectivities. The discovery of these new architectures reveals a structural versatility for this class of materials that exceeds initial expectations and observations. Each topology produces different connectivities between the sheets in the third dimension that endows each framework isomer with uniquely shaped and sized inclusion cavities, enabling this host system to conform readily to different guests. The unlimited number of architectures available, combined with the inherent conformational softness and structural tunability of these host lattices, suggests a near universality for the GS system with respect to guest inclusion.

Development of intestinal intraepithelial lymphocytes, NK cells, and NK 1.1+ T cells in CD45-deficient mice.

The transmembrane protein tyrosine phosphatase CD45 is differentially required for the development and function of B, T, and NK cells, with mice partially deficient for CD45 having a significant inhibition of T cell, but not NK or B cell, development. CD45-mediated signaling has also been implicated in the development of intrathymic, but not extrathymic, intestinal intraepithelial T lymphocytes (iIELs) in the CD45ex6(-/-) mouse. As NK1.1(+) CD3(+) (NK-T) cells can also develop through extrathymic pathways, we have investigated the role of CD45 in NK-T cell development. In mice with a complete absence of CD45 expression (CD45ex9(-/-)) the NK-T cell population was maintained in the iIEL compartment, but not in the spleen. Functionally, CD45-deficient NK-T cells were unable to secrete IL-4 in response to TCR-mediated signals, a phenotype similar to that of CD45-deficient iIELs, in which in vitro cytokine production was dramatically reduced. Using the CD45ex9(-/-) mouse strain, we have also demonstrated that only one distinct population of NK-T cells (CD8(+)) appears to develop normally in the absence of CD45. Interestingly, although an increase in cytotoxic NK cells is seen in the absence of CD45, these NK calls are functionally unable to secrete IFN-gamma. In the absence of CD45, a significant population of extrathymically derived CD8alphaalpha(+) iIELs is also maintained. These results demonstrate that in contrast to conventional T cells, CD45 is not required during the development of CD8(+) NK-T cells, NK cells, or CD8alphaalpha(+) iIELs, but is essential for TCR-mediated function and cytokine production.

Detailed methods for the quantification of nitric oxide in aqueous solutions using either an oxygen monitor or EPR.

The interest in nitric oxide has grown with the discovery that it has many biological functions. This has heightened the need for methods to quantify nitric oxide. Here we report two separate methods for the quantification of aqueous stock solutions of nitric oxide. The first is a new method based on the reaction of nitric oxide with oxygen in liquid phase (*NO + O2 + 2H2O --> 4HNO2); an oxygen monitor is used to measure the consumption of oxygen by nitric oxide. This method offers the advantages of being both simple and direct. The presence of nitrite or nitrate, frequent contaminants in nitric oxide stock solutions, does not interfere with the quantification of nitric oxide. Measuring the disappearance of dissolved oxygen, a reactant, in the presence of known amounts of nitric oxide has provided verification of the 4:1 stoichiometry of the reaction. The second method uses electron paramagnetic resonance spectroscopy (EPR) and the nitric oxide trap [Fe2+-(MGD)2], (MGD = N-methyl-D-glucamine dithiocarbamate). The nitrosyl complex is stable and easily quantitated as a room temperature aqueous solution. These two methods are validated with Sievers 280 Nitric Oxide Analyzer and cross-checked with standards using UV-Vis spectroscopy. The practical lower limits for measuring the concentration of nitric oxide using the oxygen monitor approach and EPR are approximately 3 microM and 500 nM, respectively. Both methods provide straightforward approaches for the standardization of nitric oxide in solution.