Penicillin tolerance among Beta-hemolytic streptococci and production of the group carbohydrates, hemolysins, hyaluronidases and deoxyribonucleases

Penicillin tolerance among 67 strains of beta-hemolytic streptococci was examined by determining the ratio of the minimal bactericidal concentration to the minimal inhibitory concentration as 32 or greater. Tolerance was demonstrated in 15 group A strains and in 11,7, and 4 of groups B, C and G, respectively. Thereafter the effects of a subminimal inhibitory concentration (1/2MIC) of penicillin on the bacterial products of four tolerant and four nontolerant strains (two of each Lancefield group) were analyzed and compared. The antibiotic caused a marked increase in the expression of the group carbo-hydrates for strains of group B. Penicillin was found to reduce the cell-bound hemolysin activities of the four tolerant strains and to increase the activity of the other (free) form of nontolerant groups A, C and G hemolysins. Penicillin caused an increase in the extracellular hyaluronidase activities of one group A and groups B, C and G streptococci. With added antibiotic the production of deoxyribonuclease by tolerant groups A, C and G was greatly enhanced and that of the group B streptococcus was arrested.

Inbreeding effects on progeny sex ratio and gender variation in the gynodioecious Silene vulgaris (Caryophyllaceae).

In gynodioecious species, sex expression is generally determined through cytoplasmic male sterility genes interacting with nuclear restorers of the male function. With dominant restorers, there may be an excess of females in the progeny of self-fertilized compared with cross-fertilized hermaphrodites. Moreover, the effect of inbreeding on late stages of the life cycle remains poorly explored. Here, we used hermaphrodites of the gynodioecious Silene vulgaris originating from three populations located in different valleys in the Alps to investigate the effects of two generations of self- and cross-fertilization on sex ratio and gender variation. We detected an increase in females in the progeny of selfed compared with outcrossed hermaphrodites and inbreeding depression for female and male fertility. Male fertility correlated positively with sex ratio differences between outbred and inbred progeny, suggesting that dominant restorers are likely to influence male fertility qualitatively and quantitatively in S. vulgaris. We argue that the excess of females in the progeny of selfed compared with outcrossed hermaphrodites and inbreeding depression for gamete production may contribute to the maintenance of females in gynodioecious populations of S. vulgaris because purging of the genetic load is less likely to occur.

Exploration of the contribution of isotope ratio mass spectrometry to the investigation of explosives: A study of black powders and ammonium nitrate fertilisers

The increasing number of bomb attacks involving improvised explosive devices, as well as the nature of the explosives, give rise to concern among safety and law enforcement agencies. The substances used in explosive charges are often everyday products diverted from their primary licit applications. Thus, reducing or limiting their accessibility for prevention purposes is difficult. Ammonium nitrate, employed in agriculture as a fertiliser, is used worldwide in small and large homemade bombs. Black powder, dedicated to hunting and shooting sports, is used illegally as a filling in pipe bombs causing extensive damage. If the main developments of instrumental techniques in explosive analysis have been constantly pushing the limits of detection, their actual contribution to the investigation of explosives in terms of source discrimination is limited. Forensic science has seen the emergence of a new technology, isotope ratio mass spectrometry (IRMS), that shows promising results. Its very first application in forensic science dates back to 1979. Liu et al. analysed cannabis plants coming from different countries [Liu et al. 1979]. This preliminary study highlighted its potential to discriminate specimens coming from different sources. Thirty years later, the keen interest in this new technology has given rise to a flourishing number of publications in forensic science. The countless applications of IRMS to a wide range of materials and substances attest to its success and suggest that the technique is ready to be used in forensic science. However, many studies are characterised by a lack of methodology and fundamental data. They have been undertaken in a top-down approach, applying this technique in an exploratory manner on a restricted sampling. This manner of procedure often does not allow the researcher to answer a number of questions, such as: do the specimens come from the same source, what do we mean by source or what is the inherent variability of a substance? The production of positive results has prevailed at the expense of forensic fundamentals. This research focused on the evaluation of the contribution of the information provided by isotopic analysis to the investigation of explosives. More specifically, this evaluation was based on a sampling of black powders and ammonium nitrate fertilisers coming from known sources. Not only has the methodology developed in this work enabled us to highlight crucial elements inherent to the methods themselves, but also to evaluate both the longitudinal and transversal variabilities of the information. First, the study of the variability of the profile over time was undertaken. Secondly, the variability of black powders and ammonium nitrate fertilisers within the same source and between different sources was evaluated. The contribution of this information to the investigation of explosives was then evaluated and discussed. --------------------------------------------------------------------------------------------------- Le nombre croissant d'attentats à la bombe impliquant des engins explosifs artisanaux, ainsi que la nature des charges explosives, constituent une préoccupation majeure pour les autorités d'application de la loi et les organismes de sécurité. Les substances utilisées dans les charges explosives sont souvent des produits du quotidien, détournés de leurs applications licites. Par conséquent, réduire ou limiter l'accessibilité de ces produits dans un but de prévention est difficile. Le nitrate d'ammonium, employé dans l'agriculture comme engrais, est utilisé dans des petits et grands engins explosifs artisanaux. La poudre noire, initialement dédiée à la chasse et au tir sportif, est fréquemment utilisée comme charge explosive dans les pipe bombs, qui causent des dommages importants. Si les développements des techniques d'analyse des explosifs n'ont cessé de repousser les limites de détection, leur contribution réelle à l'investigation des explosifs est limitée en termes de discrimination de sources. Une nouvelle technologie qui donne des résultats prometteurs a fait son apparition en science forensique: la spectrométrie de masse à rapport isotopique (IRMS). Sa première application en science forensique remonte à 1979. Liu et al. ont analysé des plants de cannabis provenant de différents pays [Liu et al. 1979]. Cette étude préliminaire, basée sur quelques analyses, a mis en évidence le potentiel de l'IRMS à discriminer des spécimens provenant de sources différentes. Trente ans plus tard, l'intérêt marqué pour cette nouvelle technologie en science forensique se traduit par un nombre florissant de publications. Les innombrables applications de l'IRMS à une large gamme de matériaux et de substances attestent de son succès et suggèrent que la technique est prête à être utilisée en science forensique. Cependant, de nombreuses études sont caractérisées par un manque de méthodologie et de données fondamentales. Elles ont été menées sans définir les hypothèses de recherche et en appliquant cette technique de façon exploratoire sur un échantillonnage restreint. Cette manière de procéder ne permet souvent pas au chercheur de répondre à un certain nombre de questions, tels que: est-ce que deux spécimens proviennent de la même source, qu'entend-on par source ou encore quelle est l'intravariabilité d'une substance? La production de résultats positifs a prévalu au détriment des fondamentaux de science forensique. Cette recherche s'est attachée à évaluer la contribution réelle de l'information isotopique dans les investigations en matière d'explosifs. Plus particulièrement, cette évaluation s'est basée sur un échantillonnage constitué de poudres noires et d'engrais à base de nitrate d'ammonium provenant de sources connues. La méthodologie développée dans ce travail a permis non seulement de mettre en évidence des éléments cruciaux relatifs à la méthode d'analyse elle-même, mais également d'évaluer la variabilité de l'information isotopique d'un point de vue longitudinal et transversal. Dans un premier temps, l'évolution du profil en fonction du temps a été étudiée. Dans un second temps, la variabilité du profil des poudres noires et des engrais à base de nitrate d'ammonium au sein d'une même source et entre différentes sources a été évaluée. La contribution de cette information dans le cadre des investigations d'explosifs a ensuite été discutée et évaluée.

Connexin40 regulates renin production and blood pressure.

Renin secretion is regulated by coordinated signaling between the various cells of the juxtaglomerular apparatus. The renin-secreting cells (RSC), which play a major role in the control of blood pressure, are coupled to each other and to endothelial cells by Connexin40 (Cx40)-containing channels. In this study, we show that Cx40 knockout (Cx40-/-) mice, but not their heterozygous littermates, are hypertensive due to the increase in the number of RSC, renin biosynthesis, and plasma renin. Treatment with the angiotensin II receptor AT1 antagonist candesartan or the angiotensin II-converting enzyme inhibitor ramipril reduced the blood pressure of the Cx40-/- mice to the same levels seen in wild-type (WT) mice. The elevated blood pressure of the knockout mice was not affected by clipping one renal artery (2K1C, renin-dependent model of hypertension) or after a high salt diet. Under these conditions, however, Cx40-/- mice showed an altered production and release of renin. The renin mRNA ratio between the clipped and the non-clipped kidney was lower in the knockout than in the WT 2K1C mice. This indicates that the response to a change in blood pressure was altered. The RSC of the Cx40-/- mice did not have a compensatory increase in the levels of either Cx43 or Cx37. Our data show that renin secretion is dependent on Cx40 and suggest the Cx40-/- mice may be a genetic model of renin-dependent hypertension.

Control of brood male production in the Argentine ant Iridomyrmex humilis (Mayr)

The influence of various social factors on the production of males was investigated in the Argentine ant, Iridomyrmex humilis. In this polygynous species, the workers which are monomorphic are unable to lay reproductive eggs, so all the males are the progeny of the queens. Although male eggs appear to be laid by mated queens throughout the year, in large stock colonies males are reared periodically (every 3 or 4 months); males develop from brood taken from these colonies at any point in the cycle and given queenless or queenright (1 to 5 queens) units. This is in striking contrast to many other species of ants where it is generally assumed that male eggs are laid seasonnally. Comparative experiments suggest that several related factors influence the rearing of males as far as the pupal stage. Worker/larva ratio: The proportion of male larvae developing in standardized units in which the worker/larva ratio was varied from 0.25 to 25 demonstrated that low ratios inhibit male production. Queen influence: In standardized units where the worker/larva ratio was high the presence of queens did not inhibit the rearing of males suggesting that there is no queen inhibitory pheromone controlling male experimental production. Data suggest evidence that queens prevent male production by means of appropriation of food. Diet: Male larvae failed to pupate in experimental societies deprived of protein. Thus, the production of males appears to be controlled by the amount of food available to larvae. This depends on foraging activity, the quantity of brood in relation to the number of workers and the number of queens in the society.

CO2 production of the chick embryo during the first day of post-laying development.

The carbon dioxide production of the chick embryo cultured in vitro has been determined during the first 24 h of post-laying development using a non-invasive conductometric microtechnique. The mean CO2 production of the whole blastoderm (1) increased from 16 nmol/h at laying to 231 nmol/h at early neurulation, (2) became dependent on exogenous glucose and (3) was closely linked to mechanical tension generated in the blastoderm (loosening from vitelline membrane resulted in a decrease of 56%). In our experimental conditions, no significant influence of carbonic anhydrase on the CO2 production has been detected. The value of the respiratory exchange ratio varied from about 3 at pregastrular stages to 1 at neurula stage and CO2 was produced transiently in presence of antimycin A. Such results indicate that the source of CO2 is not exclusively mitochondrial and that the relative proportions of mitochondrial and non-mitochondrial CO2 productions might vary significantly throughout the early development. Our findings confirm that the metabolism of the chick embryo becomes more and more oxidative from laying onwards and suggest that the modifications of metabolism observed during the studied period of development could be associated with functional differentiation.

Queen-worker conflict over sex ratio: a comparison of primary and secondary sex ratios in the Argentine ant,

We compare the primary sex ratio (proportion o haploid eggs laid by queens) and the secondary sex ratio (proportion of male pupae produced) in the Argentine ant Iridomyrmex humilis with the aim of investigating whether workers control the secondary sex ratio by selectively eliminating male brood. The proportion of haploid eggs produced by queens was close to 0.5 in late winter, decreased to less than 0.3 in spring and summer, and increased again to a value close to 0.5 in fall. Laboratory experiments indicate that temperture is a proximate factor influencing the primary sex ratio with a higher proportion of haploid eggs being laid at colder temperatures. Production of queen pupae ceased in mid-June, about three weeks before that of male pupae. After this time only worker pupae were produced. During the period of production of sexuals, the proportion of male pupae ranged from 0.30 to 0.38. Outside this period no males were reared although haploid eggs were produced all the year round by queens. Workers thus exert a control on the secondary sex ratio by eliminating a proportion of the male brood during the period of sexual production and eliminating all the males during the remainder of the cycle. These data are consistent with workers preferring a more female-biased sex ratio than queens. The evolutionary significance of the production of male eggs by queens all the year round is as yet unclear. It may be a mechanism allowing queen replacement in the case of the death of the queens in the colony.

Patterns of split sex ratio in ants have multiple evolutionary causes based on different within-colony conflicts.

Split sex ratio-a pattern where colonies within a population specialize in either male or queen production-is a widespread phenomenon in ants and other social Hymenoptera. It has often been attributed to variation in colony kin structure, which affects the degree of queen-worker conflict over optimal sex allocation. However, recent findings suggest that split sex ratio is a more diverse phenomenon, which can evolve for multiple reasons. Here, we provide an overview of the main conditions favouring split sex ratio. We show that each split sex-ratio type arises due to a different combination of factors determining colony kin structure, queen or worker control over sex ratio and the type of conflict between colony members.

Production of Pseudomonas aeruginosa Intercellular Small Signaling Molecules in Human Burn Wounds

Pseudomonas aeruginosa has developed a complex cell-to-cell communication system that relies on low-molecular weight excreted molecules to control the production of its virulence factors. We previously characterized the transcriptional regulator MvfR, that controls a major network of acute virulence functions in P. aeruginosa through the control of its ligands, the 4-hydroxy-2-alkylquinolines (HAQs)-4-hydroxy-2-heptylquinoline (HHQ) and 3,4-dihydroxy-2-heptylquinoline (PQS). Though HHQ and PQS are produced in infected animals, their ratios differ from those in bacterial cultures. Because these molecules are critical for the potency of activation of acute virulence functions, here we investigated whether they are also produced during human P. aeruginosa acute wound infection and whether their ratio is similar to that observed in P. aeruginosa-infected mice. We found that a clinically relevant P. aeruginosa isolate produced detectable levels of HAQs with ratios of HHQ and PQS that were similar to those produced in burned and infected animals, and not resembling ratios in bacterial cultures. These molecules could be isolated from wound tissue as well as from drainage liquid. These results demonstrate for the first time that HAQs can be isolated and quantified from acute human wound infection sites and validate the relevance of previous studies conducted in mammalian models of infection.

Carbon isotope stratigraphy and carbonate production during the Early-Middle Jurassic: examples from the Umbria-Marche-Sabina Apennines (Central Italy)

The carbon isotopic signature of carbonates depends on secular variations of organic carbon and carbonate carbon production/burial rates. A decrease in carbonate productivity makes the organic/carbonate carbon ratio unstable up to the point that even minor variations in the organic carbon reservoirs can provoke carbon isotopic shifts. The delta(13)C positive shifts of the middle Carixian (early Pliensbachian) and the early Bajocian recorded in the Umbria-Marche-Sabina domain represent a good example of this mechanism. Both sedimentology and lithostratigraphy of pelagic platform-basin carbonate systems in this area show that important changes in the source of carbonates correspond to the observed isotopic shifts. The middle Carixian event is in fact well correlatable to the drastic reduction of benthic carbonate production on rift-related intrabasinal highs, which then became pelagic carbonate platforms. The early Bajocian event is concomitant with the beginning of a long hiatus on the pelagic carbonate platforms and with a drop of the biodiversity of calcareous organisms followed by the onset of biosiliceous sedimentation in basins. (C) 2002 Elsevier Science B.V. All rights reserved.

Cholesterol metabolism is associated with soluble amyloid precursor protein production in Alzheimer's disease.

Disturbances of the cholesterol metabolism are associated with Alzheimer's disease (AD) risk and related cerebral pathology. Experimental studies found changing levels of cholesterol and its metabolites 24S-hydroxycholesterol (24S-OHC) and 27-hydroxycholesterol (27-OHC) to contribute to amyloidogenesis by increasing the production of soluble amyloid precursor protein (sAPP). The aim of this study was to evaluate the relationship between the CSF and circulating cholesterol 24S-OHC and 27-OHC, and the sAPP production as measured by CSF concentrations of sAPP forms in humans. The plasma and the CSF concentrations of cholesterol, 24S-OHC and 27-OHC, and the CSF concentrations of sAPPα, sAPPβ, and Aß1-42 were assessed in subjects with AD and controls with normal cognition. In multivariate regression tests including age, gender, albumin ratio, and apolipoprotein E (APOE)ε4 status CSF cholesterol, 24S-OHC, and 27-OHC independently predicted the concentrations of sAPPα and sAPPβ. The associations remained significant when analyses were separately performed in the AD group. Furthermore, plasma 27-OHC concentrations were associated with the CSF sAPP levels. The results suggest that high CSF concentrations of cholesterol, 24S-OHC, and 27-OHC are associated with increased production of both sAPP forms in AD.

Adjuvants that improve the ratio of antigen-specific effector to regulatory T cells enhance tumor immunity.

Antitumor immunity is strongly influenced by the balance of tumor antigen-specific effector and regulatory T cells. However, the impact that vaccine adjuvants have in regulating the balance of antigen-specific T cell populations is not well understood. We found that antigen-specific T regulatory cells (Treg) were induced following subcutaneous vaccination with either OVA or melanoma-derived peptides, with a restricted expansion of effector T cells. Addition of the adjuvants CpG-ODN or Poly(I:C) preferentially amplified effector T cells over Tregs, dramatically increasing the antigen-specific T effector:Treg ratios and inducing polyfunctional effector cells. In contrast, two other adjuvants, imiquimod and Quil A saponin, favored an expansion of antigen-specific Tregs and failed to increase effector T cell:Treg ratios. Following therapeutic vaccination of tumor-bearing mice, high ratios of tumor-specific effector T cells:Tregs in draining lymph nodes were associated with enhanced CD8+ T cell infiltration at the tumor site and a durable rejection of tumors. Vaccine formulations of peptide+CpG-ODN or Poly(I:C) induced selective production of pro-inflammatory Type I cytokines early after vaccination. This environment promoted CD8+ and CD4+ effector T cell expansion over that of antigen-specific Tregs, tipping the effector T cell to Treg balance to favor effector cells. Our findings advance understanding of the influence of different adjuvants on T cell populations, facilitating the rational design of more effective cancer vaccines.

Reduction of the bull: cow ratio in the Brazilian Pantanal

This work was conducted to study alternatives for reduction of the bull:cow ratio in the Brazilian lowland and, therefore, lower the production costs for the local beef cattle industry. The ratios 1:10, 1:25, and 1:40 were used in native pastures with a mean stocking rate of 0.27 mature animal unit per hectare over two consecutive breeding seasons. Statistical analysis did not show any effect (P>0.05) of year (P = 0.2097), animal category (P = 0.0773), bull:cow ratio (0.8134) on reproductive performance. However, the pregnancy rate in a multiple bull system was higher (P = 0.0228) than in the individual bull system. An evaluation of the economic impact of this management system in the extensive Lowland herds showed that at the ratio of 1:10 the bulls were sub utilized.

Production of nanometric particles in radio frequency glow discharges in mixtures of silane and methane

The formation of silicon particles in rf glow discharges has attracted attention due to their effect as a contaminant during film deposition or etching. However, silicon and silicon alloy powders produced by plasma¿enhanced chemical vapor deposition (PECVD) are promising new materials for sintering ceramics, for making nanoscale filters, or for supporting catalytic surfaces. Common characteristics of these powders are their high purity and the easy control of their stoichiometry through the composition of the precursor gas mixture. Plasma parameters also influence their structure. Nanometric powders of silicon¿carbon alloys exhibiting microstructural properties such as large hydrogen content and high surface/volume ratio have been produced in a PECVD reactor using mixtures of silane and methane at low pressure (-1 Torr) and low frequency square¿wave modulated rf power (13.56 MHz). The a¿Si1¿xCx:H powders were obtained from different precursor gas mixtures, from R=0.05 to R=9, where R=[SiH4]/([SiH4]+[CH4]). The structure of the a¿Si1¿xCx:H powder was analyzed by several techniques. The particles appeared agglomerated, with a wide size distribution between 5 and 100 nm. The silane/methane gas mixture determined the vibrational features of these powders in the infrared. Silicon-hydrogen groups were present for every gas composition, whereas carbon¿hydrogen and silicon¿carbon bonds appeared in methane¿rich mixtures (R-0.6). The thermal desorption of hydrogen revealed two main evolutions at about 375 and 660¿°C that were ascribed to hydrogen bonded to silicon and carbon, respectively. The estimated hydrogen atom concentration in the sample was about 50%.

Effects of nutrients and turbulence on the production of transparent exopolymer particles: a mesocosm study

The production of transparent exopolymer particles (TEP) in response to several environmental variables was studied in 2 mesocosm experiments. The first (Expt 1) examined a gradient of 4 nutrient levels; the second (Expt 2) examined different conditions of silicate availability and zooplankton presence. Tanks were separated in 2 series, one subjected to turbulence and the other not influenced by turbulence. In tanks with nutrient addition, TEP were rapidly formed, with net apparent production rates closely linked to chl a growth rates, suggesting that phytoplankton cells were actively exuding TEP precursors. High nutrient availability increased the absolute concentration of TEP; however, the relative quantity of TEP produced was found to be lower, as TEP concentration per unit of phytoplankton biomass was inversely related to the initial nitrate dose. In Expt 1, an increase in TEP volume (3 to 48 µm equivalent spherical diameter) with nutrient dose was observed; in Expt 2, both silicate addition and turbulence enhanced TEP production and favored aggregation to larger TEP (>48 µm). The presence of zooplankton lowered TEP concentration and changed the size distribution of TEP, presumably by grazing on TEP or phytoplankton. For lower nutrient concentrations, the ratio of particulate organic carbon (POC) to particulate organic nitrogen (PON) followed the Redfield ratio. At higher nutrient conditions, when nutrients were exhausted during the post-bloom, a decoupling of carbon and nitrogen dynamics occurred and was correlated to TEP formation, with a large flow of carbon channeled toward the TEP pool in turbulent tanks. TEP accounted for an increase in POC concentration of 50% in high-nutrient and turbulent conditions. The study of TEP dynamics is crucial to understanding the biogeochemical response of the aquatic system to forcing variables such as nutrient availability and turbulence intensity.