Staphylococcus aureus forms spreading dendrites that have characteristics of active motility

Staphylococcus aureus is historically regarded as a non-motile organism. More recently it has been shown that S. aureus can passively move across agar surfaces in a process called spreading. We re-analysed spreading motility using a modified assay and fo- cused on observing the formation of dendrites: branching structures that emerge from the central colony. We discovered that S. aureus can spread across the surface of media in struc- tures that we term ‘comets’, which advance outwards and precede the formation of dendrites. We observed comets in a diverse selection of S. aureus isolates and they exhibit the following behaviours: (1) They consist of phenotypically distinct cores of cells that move forward and seed other S. aureus cells behind them forming a comet ‘tail’; (2) they move when other cells in the comet tail have stopped moving; (3) the comet core is held together by a matrix of slime; and (4) the comets etch trails in the agar as they move forwards. Comets are not con- sistent with spreading motility or other forms of passive motility. Comet behaviour does share many similarities with a form of active motility known as gliding. Our observations therefore suggest that S. aureus is actively motile under certain conditions

Bt protein rhizosecreted from transgenic maize does not accumulate in soil

The persistence of CryIAb protein rhizosecreted in soil is important in the assessment of its environmental risk. Here we report that CryIAb protein from transgenic maize does not accumulate at high levels in soils. Levels of CryIAb protein rhizosecreted by three maize transgenic events (BT11, MON810 and 176) were studied in hydroponic cultures and found only in the MON810 and BT11 events but not in event 176 or control plants. Under field conditions, the cryIAb gene and a basal level of CryIAb protein was detected in soils from plots cultivated with transgenic and non-transgenic maize, possibly from Bacillus thuringiensis present in the soils

Small molecule binding sites on the Ras:SOS complex can be exploited for inhibition of Ras activation.

Constitutively active mutant KRas displays a reduced rate of GTP hydrolysis via both intrinsic and GTPase-activating protein-catalyzed mechanisms, resulting in the perpetual activation of Ras pathways. We describe a fragment screening campaign using X-ray crystallography that led to the discovery of three fragment binding sites on the Ras:SOS complex. The identification of tool compounds binding at each of these sites allowed exploration of two new approaches to Ras pathway inhibition by stabilizing or covalently modifying the Ras:SOS complex to prevent the reloading of Ras with GTP. Initially, we identified ligands that bound reversibly to the Ras:SOS complex in two distinct sites, but these compounds were not sufficiently potent inhibitors to validate our stabilization hypothesis. We conclude by demonstrating that covalent modification of Cys118 on Ras leads to a novel mechanism of inhibition of the SOS-mediated interaction between Ras and Raf and is effective at inhibiting the exchange of labeled GDP in both mutant (G12C and G12V) and wild type Ras

Defining motility in the Staphylococci

The ability of bacteria to move is critical for their survival in diverse environments and multiple ways have evolved to achieve this. Two forms of motility have recently been described for Staphylococcus aureus, an organism previously considered to be non-motile. One form is called spreading, which is a type of sliding motility and the second form involves comet formation, which has many observable characteristics associated with gliding motility. Darting motility has also been observed in Staphylococcus epidermidis. This review describes how motility is defined and how we distinguish between passive and active motility. We discuss the characteristics of the various forms of Staphylococci motility, the molecular mechanisms involved and the potential future research directions

Adenocarcinoma in Caroli's Disease Treated by Liver Transplantation

Caroli's disease is characterized by congenital cystic dilatation of the intrahepatic bile ducts. In 7% of casea a malignant tumor develops complicating the course of the disease

El acondicionamiento con agua caliente afecta diferencialmente el metabolismo de carbohidratos y la producción de volátiles en la poscosecha de ‘Murcott’ y ‘W-Murcott’ (Citrus reticulata blanco)

La producción citrícola es una de las actividades comerciales más relevantes de nuestro país y del mundo. La conservación a bajas temperaturas para evitar su decaimiento, suele provocar mermas en su calidad, por lo cual es frecuente la aplicación de diversos tratamientos de acondicionamiento. El objetivo del trabajo fue comparar la incidencia del tratamiento con agua caliente sobre la producción de volátiles y la actividad de las principales enzimas del metabolismo de carbohidratos en dos variedades de mandarinas, ‘Murcott’ y ‘W-Murcott’, así como su impacto en la poscosecha. Las mandarinas recién cosechadas fueron tratadas con agua caliente (HWT) a 50 °C por 3 min, mientras que la fruta control (C) se sumergió 3 min en agua a 15 °C. Se almacenaron a 5 °C durante 33 días, a continuación, se mantuvieron por 7 días más a 20 °C, para simular el período de comercialización. Se tomaron muestras antes de tratar (T0) y a 1, 9 y 33 + 7 días. Se realizó la determinación de acetaldehído, etanol y pérdida de peso por métodos estándar. En muestras de flavedo se determinó el contenido de metabolitos primarios por GC-MS y las actividades de las enzimas ATP-PFK; PPi-PFK; PK; MDH; AlaT; HK; PGI; PEPC y Glu6-P DH. La disminución de peso seco, la producción de etanol y de acetaldehído fueron mayores en ‘W-Murcott’, siendo menor la pérdida de peso durante la poscosecha en frutas tratadas. En estas mismas muestras, los niveles de ambos volátiles superaron el valor tolerado (1500-2000 ppm) y los de citrato fueron menores. El análisis de las variaciones en los metabolitos y las enzimas, evidenció un mayor efecto general del tratamiento en ‘W-Murcot’, mientras ‘Murcott’ se vio más afectada por el almacenamiento. La conversión de fru-6-P a fru-1,6-BP es catalizada en plantas por la ATP-PFK (irreversible) y la PPi-PFK (reversible). La PPi-PFK se induce por falta de Pi y ATP, y fue más alta en ‘W-Murcott’, sin efecto del tratamiento. En esta variedad, los bajos niveles de citrato y de actividades HK y PGI, favorecerían la gluconeogénesis en las frutas tratadas. Las fluctuaciones analizadas de algunas enzimas y metabolitos, más notorias en la fruta tratada de ‘W-Murcott’, son compatibles con diversos destinos del carbono que favorecerían el mantenimiento de la calidad y la protección frente a diversos tipos de estrés en esta variedad. ‘W- Murcott’ parece sufrir mayor estrés durante la poscosecha que ‘Murcott’, incrementado por el tratamiento térmico en cuanto a ciertos parámetros. Las actividades de muchas de las enzimas determinadas mostraron mayor influencia de la variedad que del tratamiento

Group B Streptococcus vaccine development: present status and future considerations, with emphasis on perspectives for low and middle income countries.

Globally, group B Streptococcus (GBS) remains the leading cause of sepsis and meningitis in young infants, with its greatest burden in the first 90 days of life. Intrapartum antibiotic prophylaxis (IAP) for women at risk of transmitting GBS to their newborns has been effective in reducing, but not eliminating, the young infant GBS disease burden in many high income countries. However, identification of women at risk and administration of IAP is very difficult in many low and middle income country (LMIC) settings, and is not possible for home deliveries. Immunization of pregnant women with a GBS vaccine represents an alternate pathway to protecting newborns from GBS disease, through the transplacental antibody transfer to the fetus in utero. This approach to prevent GBS disease in young infants is currently under development, and is approaching late stage clinical evaluation. This manuscript includes a review of the natural history of the disease, global disease burden estimates, diagnosis and existing control options in different settings, the biological rationale for a vaccine including previous supportive studies, analysis of current candidates in development, possible correlates of protection and current status of immunogenicity assays. Future potential vaccine development pathways to licensure and use in LMICs, trial design and implementation options are discussed, with the objective to provide a basis for reflection, rather than recommendations

Proteomic and metabolomic approaches unveil relevant biochemical changes in carbohydrate and cell wall metabolisms of two blueberry (Vaccinium corymbosum) varieties with different quality attributes

Quality maintenance in rapidly decaying fruit such as blueberries (Vaccinium corymbosum) is of essential importance to guarantee the economic success of the crop. Fruit quality is a multifaceted subject that encompasses flavor, aroma, visual and physical issues as main factors. In this paper we report an ample characterization of different biochemical and physical aspects in two varieties (O'Neal and Emerald) of blueberries that differ in firmness, aspect, flavor and harvesting times, at two different phenological stages (fruit set vs. ripe), with the intention of unveiling how the metabolic signature of each contributes to their contrasting quality. To this effect a metabolomic, ionomic and proteomic approach was selected. The results presented here show marked differences in several variables at the two stages and between varieties. Emerald is an early variety with a large, good taste and firm fruit, while O'Neal is soft, medium sized and very sweet. Proteomic data comparison between both cultivars showed that, at fruit set, processes related with the response to inorganic compounds and small molecule metabolisms are relevant in both varieties. However, solute accumulation (mainly amino acids and organic acids), enzymes related with C: N balance, water transport and cell wall recycling are enhanced in Emerald. In ripe fruit, Emerald showed an enrichment of proteins associated with TCA, nitrogen, small molecules and cell wall in muro recycling processes, while mannitol and fatty acid metabolism were enhanced in the soft variety. The measured variation in metabolite levels gave strong support to the precedent results. This study suggests that at fruit set, a composite scenario of active metabolic recycling of the cell wall, improved C: N balance and solute accumulation give place to a more efficient carbon and water resource management. During the ripe stage, an increased and efficient in muro and metabolic recycling of the cell wall, added to enhanced inositol and secondary metabolism may be responsible for a best turgor conservation in Emerald. These findings may yield clues for improvements in fertilization practices, as well as to assist the guided development of new varieties based on biochemical quality.Fil: Montecchiarini, Marina Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Margarit, Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Morales, Luisina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Rivadeneira, María Fernanda. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concordia; ArgentinaFil: Bello, Fernando. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concordia; ArgentinaFil: Gollán, A.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concordia; ArgentinaFil: Vázquez, Daniel E.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concordia; ArgentinaFil: Podesta, Florencio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Tripodi, Karina Eva Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentin

Light inhibition of foliar respiration in response to soil water availability and seasonal changes in temperature Mediterranean holm oak. (quercus ilex forest)

In the present study we investigated variations in leaf respiration in darkness (RD) and light (RL), and associated traits in response to season, and along a gradient of soil moisture, in Mediterranean woodland dominated by holm oak (Quercus ilex L.) in central and north-eastern Spain respectively. On seven occasions during the year in the central Spain site, and along the soil moisture gradient in north-eastern Spain, we measured rates of leaf RD, RL (using the Kok method), light-saturated photosynthesis (A) and related light response characteristics, leaf mass per unit area (MA) and leaf nitrogen (N) content. At the central Spain site, significant seasonal changes in soil water content and ambient temperature (T) were associated with changes in MA, foliar N, A and stomatal conductance. RD measured at the prevailing daily T and in instantaneous R-T responses, displayed signs of partial acclimation and was not significantly affected by time of year. RL was always less than, and strongly related to, RD, and RL/RD did not vary significantly or systematically with seasonal changes in T or soil water content. Averaged over the year, RL/RD was 0.66 ± 0.05 s.e. (n = 14) at the central Spain site. At the north-eastern Spain site, the soil moisture gradient was characterised by increasing MA and RD, and reduced foliar N, A, and stomatal conductance as soil water availability decreased. Light inhibition of R occurred across all sites (mean RL/RD = 0.69 ± 0.01 s.e. (n = 18)), resulting in ratios of RL/A being lower than for RD/A. Importantly, the degree of light inhibition was largely insensitive to changes in soil water content. Our findings provide evidence for a relatively constrained degree of light inhibition of R (RL/RD ~ 0.7, or inhibition of ~30%) across gradients of water availability, although combined impacts of seasonal changes in both T and soil water content increase the range of values expressed. The findings thus have implications in terms of the assumptions made by predictive models that seek to account for light inhibition of R, and for our understanding of how environmental gradients impact on leaf trait relationships in Mediterranean plant communities

Genomic and expression analysis of multiple Sry loci from a single Rattus norvegicus Y chromosome

BACKGROUND: Sry is a gene known to be essential for testis determination but is also transcribed in adult male tissues. The laboratory rat, Rattus norvegicus, has multiple Y chromosome copies of Sry while most mammals have only a single copy. DNA sequence comparisons with other rodents with multiple Sry copies are inconsistent in divergence patterns and functionality of the multiple copies. To address hypotheses of divergence, gene conversion and functional constraints, we sequenced Sry loci from a single R. norvegicus Y chromosome from the Spontaneously Hypertensive Rat strain (SHR) and analyzed DNA sequences for homology among copies. Next, to determine whether all copies of Sry are expressed, we developed a modification of the fluorescent marked capillary electrophoresis method to generate three different sized amplification products to identify Sry copies. We applied this fragment analysis method to both genomic DNA and cDNA prepared from mRNA from testis and adrenal gland of adult male rats. RESULTS: Y chromosome fragments were amplified and sequenced using primers that included the entire Sry coding region and flanking sequences. The analysis of these sequences identified six Sry loci on the Y chromosome. These are paralogous copies consistent with a single phylogeny and the divergence between any two copies is less than 2%. All copies have a conserved reading frame and amino acid sequence consistent with function. Fragment analysis of genomic DNA showed close approximations of experimental with predicted values, validating the use of this method to identify proportions of each copy. Using the fragment analysis procedure with cDNA samples showed the Sry copies expressed were significantly different from the genomic distribution (testis p < 0.001, adrenal gland p < 0.001), and the testis and adrenal copy distribution in the transcripts were also significantly different from each other (p < 0.001). Total Sry transcript expression, analyzed by real-time PCR, showed significantly higher levels of Sry in testis than adrenal gland (p, 0.001). CONCLUSION: The SHR Y chromosome contains at least 6 full length copies of the Sry gene. These copies have a conserved coding region and conserved amino acid sequence. The pattern of divergence is not consistent with gene conversion as the mechanism for this conservation. Expression studies show multiple copies expressed in the adult male testis and adrenal glands, with tissue specific differences in expression patterns. Both the DNA sequence analysis and RNA transcript expression analysis are consistent with more than one copy having function and selection preventing divergence although we have no functional evidence