The Sodium/Proline Transporter PutP of Helicobacter pylori

Helicobacter pylori is cause of chronic gastritis, duodenal ulcer and gastric carcinoma in humans. L-proline is a preferred energy source of the microaerophilic bacterium. Previous analyses revealed that HpputP and HpputA, the genes that are predicted to play a central role in proline metabolism as they encode for the proline transporter and proline dehydrogenase, respectively, are essential for stomach colonization. Here, the molecular basis of proline transport in H. pylori by HpPutP was investigated experimentally for the first time. Measuring radiolabeled substrate transport in H. pylori and E. coli heterologously expressing HpputP as well as in proteoliposomes reconstituted with HpPutP, we demonstrate that the observed proline transport in H. pylori is mediated by HpPutP. HpPutP is specific and exhibits a high affinity for L-proline. Notably, L-proline transport is exclusively dependent on Na⁺ as coupling ion, i.e., Na⁺/L-proline symport, reminiscent to the properties of PutP of E. coli even though H. pylori lives in a more acidic environment. Homology model-based structural comparisons and substitution analyses identified amino acids crucial for function. HpPutP-catalyzed proline uptake was efficiently inhibited by the known proline analogs 3,4-dehydro-D,L-proline and L-azetidine-2-carboxylic acid.     

The thermoregulatory benefits of the communal nest of sociable weavers Philetairus socius are spatially structured within nests

Structures built by animals, such as nests, mounds and burrows, are often the product of cooperative investment by more than one individual. Such structures may be viewed as a public good, since all individuals that occupy them share the benefits they provide. However, access to the benefits generated by the structure may vary among individuals and is likely to be an important determinant of social organisation. Here we use the massive, communal nests of sociable weavers Philetairus socius, to investigate whether their thermoregulatory function varies in relation to the size of communal nests, and the position of individual nest chambers within the communal structure. We then examine whether this spatial variation in thermoregulatory function predicts the social organisation of colonies. First, we show that the sociable weavers’ communal nests buffer variation in ambient temperature, and reduce temperature variability within nest chambers. The extent of this buffering effect depends significantly on the position of nest chambers within the communal structure, and on the depth to which chambers are embedded within the nest mass. We detected no effect of nest volume on thermoregulatory benefits, suggesting that there are likely to be additional, non‐thermoregulatory benefits leading to communal nests. Finally, our results indicate that there may be competition for access to the benefits of the public good, since older birds occupied the chambers with the highest thermoregulatory benefits, where breeding activity was also more common. We discuss how the spatial structure of the benefits of the public good might influence social organisation in the unique communal lifestyle of sociable weavers.     

C-kit mutations and PKC crosstalks: PKC translocates to nucleous only in cells HMC⁵⁶⁰,⁸¹⁶

The human mast cell lines HMC‐1⁵⁶⁰ and HMC‐1^560,816 were used to study histamine release, Ca²⁺ signaling and protein kinase C (PKC) localization and expression, with phorbol 12‐myristate 13‐acetate (PMA). Both sublines carry activating mutations in the proto‐oncogene of c‐kit that cause autophosphorylation and permanent c‐kit tyrosine kinase activation. Both have the Gly‐560 → Val mutation but only the second carries the Asp‐816 → Val mutation. In this study, it was observed that the stimulation of PKC has different effects in HMC‐1⁵⁶⁰ and HMC‐1^560,816 and this would be related to the difference in activating mutations in both mast cell lines. PKC activation increases ionomycin‐induced histamine release in HMC‐1⁵⁶⁰. This article demonstrates an opposite histamine response in HMC‐1^560,816 cells, even though classical PKCs are the family of isozymes responsible for this effect in both cellular lines. Furthermore, it can be observed that upon cell stimulation with PMA, primarily cytosolic PKC translocates to the nucleous in HMC‐1^560,816 cells, but not in HMC‐1⁵⁶⁰ cell line. J. Cell. Biochem. 112: 2637–2651, 2011. © 2011 Wiley‐Liss, Inc.     

Caveolin-1 regulates cell polarization and directional migration through Src kinase and Rho GTPases

Development, angiogenesis, wound healing, and metastasis all involve the movement of cells in response to changes in the extracellular environment. To determine whether caveolin-1 plays a role in cell migration, we have used fibroblasts from knockout mice. Caveolin-1–deficient cells lose normal cell polarity, exhibit impaired wound healing, and have decreased Rho and increased Rac and Cdc42 GTPase activities. Directional persistency of migration is lost, and the cells show an impaired response to external directional stimuli. Both Src inactivation and p190RhoGAP knockdown restore the wild-type phenotype to caveolin-1–deficient cells, suggesting that caveolin-1 stimulates normal Rho GTP loading through inactivation of the Src–p190RhoGAP pathway. These findings highlight the importance of caveolin-1 in the establishment of cell polarity during directional migration through coordination of the signaling of Src kinase and Rho GTPases.     

Genetic variability of Phytophthora sojae isolates from Argentina

Phytophthora sojae causes root and stem rot, one of the most important diseases of soybean worldwide. Genetic diversity of 32 Phytophthora sojae isolates of different geographic origin from Argentina was evaluated with RAPD markers. The isolates were collected from diseased soybean plants and soil samples from Santa Fe, Buenos Aires, C6rdoba and Entre Rios provinces, in the Pampeana Region. DNA was amplified with 20 decanucleotides primers. Seven primers amplified 49 fragments, of which 35 were polymorphic, indicating high variability. RAPD analysis detected intraspecific variability even among isolates of the same geographic origin.     

Interannual variations in feeding frequencies and food quality of greater flamingo chicks (Phoenicopterus roseus): evidence from plasma chemistry and effects on body condition

Greater flamingos in southern Spain foraged in areas distant from a breeding site, spending 4-6 days in foraging areas between successive visits to the colony to feed their chicks. During four years, we took blood samples from chicks to ascertain whether there were interannual variations in several blood parameters, indicative of food quality and feeding frequencies. When the chicks were captured, 20-31% of them had their crops empty, indicating that not all chicks were fed daily. Additional evidence of variations in feeding frequencies was obtained from a principal component analysis (PCA) on plasma chemistry values, which also indicated that there were annual variations in the quality of food received by chicks. The association of cholesterol and glucose with some PC axes indicated that some chicks were experiencing fasting periods. Of all plasma metabolites considered, cholesterol was the best one to predict body condition. Greater flamingo chicks experiencing longer fasting intervals, as suggested by higher plasma levels of cholesterol, were in lower body condition.     

Guidelines for diagnosis and treatment of Hunter Syndrome for clinicians in Latin America

This review aims to provide clinicians in Latin America with the most current information on the clinical aspects, diagnosis, and management of Hunter syndrome, a serious and progressive disease for which specific treatment is available. Hunter syndrome is a genetic disorder where iduronate-2-sulfatase (I2S), an enzyme that degrades glycosaminoglycans, is absent or deficient. Clinical manifestations vary widely in severity and involve multiple organs and tissues. An attenuated and a severe phenotype are recognized depending on the degree of cognitive impairment. Early diagnosis is vital for disease management. Clinical signs common to children with Hunter syndrome include inguinal hernia, frequent ear and respiratory infections, facial dysmorphisms, macrocephaly, bone dysplasia, short stature, sleep apnea, and behavior problems. Diagnosis is based on screening urinary glycosaminoglycans and confirmation by measuring I2S activity and analyzing I2S gene mutations. Idursulfase (recombinant I2S) (Elaprase^®, Shire) enzyme replacement therapy (ERT), designed to address the underlying enzyme deficiency, is approved treatment and improves walking capacity and respiratory function, and reduces spleen and liver size and urinary glycosaminoglycan levels. Additional measures, responding to the multi-organ manifestations, such as abdominal/inguinal hernia repair, carpal tunnel surgery, and cardiac valve replacement, should also be considered. Investigational treatment options such as intrathecal ERT are active areas of research, and bone marrow transplantation is in clinical practice. Communication among care providers, social workers, patients and families is essential to inform and guide their decisions, establish realistic expectations, and assess patients’ responses.     

Immune Suppressive Effect of Cinnamaldehyde Due to Inhibition of Proliferation and Induction of Apoptosis in Immune Cells: Implications in Cancer

Background

Besides its anti-inflammatory effects, cinnamaldehyde has been reported to have anti-carcinogenic activity. Here, we investigated its impact on immune cells.

Methods

Activation of nuclear factor-κB by cinnamaldehyde (0–10 µg/ml) alone or in combination with lipopolysaccharide was assessed in THP1XBlue human monocytic cell line and in human peripheral blood mononuclear cells (PBMCs). Proliferation and secretion of cytokines (IL10 and TNFα) was determined in primary immune cells and the human cell lines (THP1, Jurkat E6-1 and Raji cell lines) stimulated with cinnamaldehyde alone or in conjunction with lipopolysaccharide. Nitric oxide was determined in mouse RAW264.7 cells. Moreover, different treated PBMCs were stained for CD3, CD20 and AnnexinV.

Results

Low concentrations (up to 1 µg/ml) of cinnamaldehyde resulted in a slight increase in nuclar factor-kB activation, whereas higher concentrations led to a dose-dependent decrease of nuclear factor-kB activation (up to 50%) in lipopolysachharide-stimulated THP1 cells and PBMCs. Accordingly, nitric oxide, interleukin 10 secretion as well as cell proliferation were reduced in lipopolysachharide-stimulated RAW264.7 cells, PBMCs and THP1, Raji and Jurkat-E6 immune cells in the presence of cinnamaldehyde in a concentration-dependent manner. Flow cytometric analysis of PBMCs revealed that CD3+ were more affected than CD20+ cells to apopotosis by cinnamaldehyde.

Conclusion

We attribute the anti-inflammatory properties of cinnamaldehyde to its ability to block nuclear factor-κB activation in immune cells. Treatment with cinnamaldehyde led to inhibition of cell viability, proliferation and induced apoptosis in a dose-dependent manner in primary and immortalized immune cells. Therefore, despite its described anti-carcinogenic property, treatment with cinnamaldehyde in cancer patients might be contraindicated due to its ability to inhibit immune cell activation.