Behaviour of bacterial division protein FtsZ under a monolayer with phospholipid domains

Assembly of the tubulin-like protein FtsZ at or near the cytoplasmic membrane is one of the earliest steps in division of bacteria such as Escherichia coli. Exactly what constitutes the site at which FtsZ acts is less clear. To investigate the influence of the membrane phospholipids on FtsZ localization and assembly, we have elaborated with the Langmuir technique a two-lipid monolayer made of dilauryl-phosphatidylethanolamine (DLPE) and dipalmitoyl-phosphatidylglycerol (DPPG). This monolayer comprised stable condensed domains in an expanded continuous phase. In the presence of GTP, FtsZ assembly disrupts the condensed domains within 5 min. After several hours, with or without GTP, FtsZ assembled into large aggregates at the domain interface. We suggest that the GTP-induced polymerization of FtsZ is coupled to the association of FtsZ protofilaments with domain interfaces.     

Serum α-tocopherol and selenium in belgian infants and children

Previous studies showed low selenium (Se) concentrations in Belgian children. Serum α-tocopherol, retinol, total cholesterol, high-density lipoprotein and low-density lipoprotein cholesterol, selenium (Se), and thiobarbituric acid-reactive substances were examined. In order to obtain further information on the Se status in Belgian children, Se, α-tocopherol, retinol, and lipid concentrations were examined and signs of peroxidative lipid damage were evaluated in a subgroup. The study was performed in 524 children (0–14 yr old) during vaccination campaigns. Three age groups were analyzed: 0–1, 1–4, and 4–14 yr. In 87 of them, where sufficient amounts of serum were available, analysis of thiobarbituric acid-reactive substances was done. Infants have high serum α-tocopherol concentrations: (23.2 μmol/L [median and interquartile range: 18.6–30.2]) and low Se concentrations (0.37 mol/L [0.27–0.47]). Se concentrations rise significantly during the first 4 yr (p < 0.0001) (Mann-Whitney U-test, tied p-values): 0.70 μmol/L (0.59–0.82); in the 4–14 yr olds, it was 0.75 μmol/L (0.67–0.86). These values remain low compared to results coming from other parts of the world. α-Tocopherol concentrations decrease significantly after infancy (p < 0.0001). The ratio α-tocopherol/total cholesterol is higher in infants. This is induced by the high vitamin E content of infant formulas. Signs of serum lipid peroxidation could not be detected by analysis of serum malondialdehyde concentrations. High α-tocopherol concentrations, as those observed in infant serum lipids, could be one of the protective mechanisms from the peroxidative lipid damages, sometimes observed in a low-Se status.     

Radiologic diagnosis of urinary tract disorders in genital endometriosis]

The paper is concerned with the results of combined investigation of 157 women suffering from genital endometriosis, operated on for this disease. The involvement of the lower parts of the ureters in a pathological process was detected in 48 patients before operation and in 6 patients at operation. Comparison of x-ray and operative findings with histology specimens made it possible to define endogenous and exogenous forms of ureteral endometriosis and three variants of an x-ray picture in this pathology (the absence of abnormal changes of the urinary tract, hydroureter is over the narrowed part of the ureter, hydroureteronephrosis). The authors have shown a high informative value of combined x-ray investigation and the effectiveness of its use in clinical practice.     

Complete genome sequence of the sulfur compounds oxidizing chemolithoautotroph Sulfuricurvum kujiense type strain (YK-1(T))

Sulfuricurvum kujiense Kodama and Watanabe 2004 is the type species of the monotypic genus Sulfuricurvum, which belongs to the family Helicobacteraceae in the class Epsilonproteobacteria. The species is of interest because it is frequently found in crude oil and oil sands where it utilizes various reduced sulfur compounds such as elemental sulfur, sulfide and thiosulfate as electron donors. Members of the species do not utilize sugars, organic acids or hydrocarbons as carbon and energy sources. This genome sequence represents the type strain of the only species in the genus Sulfuricurvum. The genome, which consists of a circular chromosome of 2,574,824 bp length and four plasmids of 118,585 bp, 71,513 bp, 51,014 bp, and 3,421 bp length, respectively, harboring a total of 2,879 protein-coding and 61 RNA genes and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.     

The effects of two bouts of high- and low-volume resistance exercise on glucose tolerance in normoglycemic women

The purpose of the study was to determine the efficacy of a low-volume, moderate-intensity bout of resistance exercise (RE) on glucose, insulin, and C-peptide responses during an oral glucose tolerance test (OGTT) in untrained women compared with a bout of high-volume RE of the same intensity. Ten women (age 30.1 ± 9.0 years) were assessed for body composition, maximal oxygen uptake, and 1-repetition maximum (1RM) before completing 3 treatments administered in random order: 1 set of 10 REs (RE1), 3 sets of 10 REs (RE3), and no exercise (C). Twenty-four hours after completing each treatment, an OGTT was performed after an overnight fast. Glucose area under the curve response to an OGTT was reduced after both RE1 (900 ± 113 mmol·L(-1)·min(-1), p = 0.056) and RE3 (827.9 ± 116.3, p = 0.01) compared with C (960.8 ± 152.7 mmol·L(-1)·min(-1)). Additionally, fasting glucose was significantly reduced after RE3 (4.48 ± 0.45 vs. 4.90 ± 0.44 mmol·L(-1), p = 0.01). Insulin sensitivity (IS), as determined from the Cederholm IS index, was improved after RE1 (10.8%) and after RE3 (26.1%). The reductions in insulin and C-peptide areas after RE1 and RE3 were not significantly different from those in the C treatment. In conclusion, greater benefits in glucose regulation appear to occur after higher volumes of RE. However, observed reductions in glucose, insulin, C-peptide areas after RE1 suggest that individuals who may not well tolerate high-volume RE protocols may still benefit from low-volume RE at moderate intensity (65% 1RM).     

Alterations in glycopeptides associated with herceptin treatment of human breast carcinoma mcf-7 and T-lymphoblastoid cells

The therapeutic humanized monoclonal antibody IgG1 known as Herceptin^® has shown remarkable antitumor effects. Although this type of therapy has increased the cancer-free survival of patients, not all tumors respond to this treatment and cancers often develop resistance to the antibody. Despite the fact that Herceptin function has been extensively studied, the precise mechanism underlying its antitumor activity still remains incompletely defined. We previously demonstrated on human breast MCF-7 carcinoma and T-lymphoblastoid CEM cells that monoclonal antibody in combination with Lipoplex consisting of Lipofectamine mixed with plasmid DNA showed a more profound effect on cancer cell viability than antibody alone. The analyses of N-glycans isolated from cancer cells showed dramatic differences in profiles when cells were exposed to Herceptin. Moreover, the investigation of glycosylated peptides from the same cancer cell models after treatment revealed further alterations in the post-translational modifications. Tandem mass spectra obtained from the samples treated confirmed the presence of a series of glycopeptides bearing characteristic oligosaccharides as described in IgG1. However some of them differed by mass differences that corresponded to peptide backbones not described previously and more of them were detected from Herceptin treated samples than from cells transfected with Heceptin/Lipoplex. The results indicate that the presence of Lipoplex prevents antibody transformation and elongates its proper function. The better understanding of the multipart changes described in the glycoconjugates could provide new insights into the mechanism by which antibody induces regression in cancers.Glycosylation of proteins is a ubiquitous type of post-translational modification in living systems. Variations in oligosaccharide structures are associated with many normal and pathological events such as cellular growth, host-pathogen interaction, differentiation, migration, cell trafficking, or tumor invasion (1, 2). Targeted glycosylation research has become important in the area of developing novel therapeutic approaches (3–5). The structures of asparagine-linked oligosaccharides in the conserved C_H2 region of the constant Fc domain of human immunoglobulin-γ (IgG1) have been shown to affect the pharmacokinetics, antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity (6, 7). In the last decade, many recombinant antibody molecules have been licensed for the treatment of a variety of cancers and chronic diseases (8). Herceptin, also known as Trastuzumab, marketed by Genentech Inc. is one example of therapeutic IgG1 antibody. It is produced from mammalian cell culture using Chinese hamster ovary cells (9). The main oligosaccharide forms found in this polypeptide chain in the Fc domain at asparagine 297 are biantennary core-fucosylated complex type structures with variable terminal galactosylation (zero, one, or two galactose residues) on their nonreducing termini (10, 11). This humanized monoclonal antibody is known to effectively target breast cancer cells overexpresing the human epidermal growth factor receptor HER2/neu (12). HER2 is a cell membrane surface-bound receptor tyrosine kinase and is normally involved in the signal transduction pathways leading to cell growth and differentiation. It can be found overexpressed in a variety tumors'' cells of epithelial origin and hematological malignancies, including acute lymphoblastic leukemia (13). When antibody binds to defective HER2 protein, this protein no longer causes cells to reproduce uncontrollably. This increases the survival of people with cancer. However, cancers usually develop resistance to trastuzumab. Unfortunately, only 25–30% of patients with HER2/neu positive breast cancer respond to this antibody (14–17). Therefore search for the potential biomarkers that could predict the efficacy of clinical outcomes is needed. More precise investigation on cellular and molecular level might provide many exciting insights in understanding of mechanism resistance cancer cells to the antibody, so that antibody-based therapies can be optimized more individually (18).We recently demonstrated how the carbohydrate moieties of two cancer cell models were affected during treatment with antibody (19). The detailed glycans profiles studied by means of mass spectrometry (MS) from the two most common cancer cell lines—human breast MCF-7 carcinoma and T-lymphoblastoid CEM cells before and after treatment with Herceptin showed significant differences. Dominant high-mannose structures analyzed in both original cancer cells were suppressed after treatment and instead, complex bi- and triantennary glycans were the major structures found in the treated samples. Their ratio or occurrence varied with conditions and time of exposure of the cancer cells to the antibody. The results provided very good evidence for involvement of glycosylation during treatment. In this regard, continuous work presented here on this subject has been aimed to the MS investigation of glycosylated peptides generated by proteolytic digestions of the cancer cells before and after exposure to Herceptin or Herceptin/Lipoplex. Direct analysis of glycopeptides by tandem MS has been shown as one of the most sensitive and fast methods for a site-specific characterization of glycosylation. It can provide information on glycan composition, glycan attachment site with determination of peptide sequence (20–28), and may offer more specific biomarkers to monitor changes in the post-translational modification at the onset, during cancer progression or during treatment.     

Interaction with both ZNRF3 and LGR4 is required for the signalling activity of R‐spondin

R‐spondin proteins sensitize cells to Wnt signalling and act as potent stem cell growth factors. Various membrane proteins have been proposed as potential receptors of R‐spondin, including LGR4/5, membrane E3 ubiquitin ligases ZNRF3/RNF43 and several others proteins. Here, we show that R‐spondin interacts with ZNRF3/RNF43 and LGR4 through distinct motifs. Both LGR4 and ZNRF3 binding motifs are required for R‐spondin‐induced LGR4/ZNRF3 interaction, membrane clearance of ZNRF3 and activation of Wnt signalling. Importantly, Wnt‐inhibitory activity of ZNRF3, but not of a ZNRF3 mutant with reduced affinity to R‐spondin, can be strongly suppressed by R‐spondin, suggesting that R‐spondin primarily functions by binding and inhibiting ZNRF3. Together, our results support a dual receptor model of R‐spondin action, where LGR4/5 serve as the engagement receptor whereas ZNRF3/RNF43 function as the effector receptor.     

Rapid Growth Mutants of Escherichia Coli

If rapid growth (rap) mutants of Escherichia coli could be obtained, these might prove a valuable contribution to fields as diverse as growth rate control, biotechnology and the regulation of the bacterial cell cycle. To obtain rap mutants, a dnaQ mutator strain was grown for four and a half days continuously in batch culture. At the end of the selection period, there was no significant change in growth rate. This result means that selecting rap mutants may require an alternative strategy and a number of such alternatives are discussed.     

Anti-Predator Benefits of Mixed-Species Groups of Cowtail Stingrays (Pastinachus sephen) and Whiprays (Himantura uarnak) at Rest

Heterospecific grouping can sometimes provide greater antipredator benefits to individuals than grouping with conspecifics. We explored the potential benefits of mixed‐species group resting in the cowtail stingray, Pastinachus sephen, and the reticulate whipray, Himantura uarnak, in Shark Bay, Western Australia. From focal follow data on individual resting choice, we first ascertained that cowtails preferred to rest with heterospecifics, as they chose to settle next to whiprays more often than to pass them (with the opposite trend observed for conspecifics). In addition, we determined from filmed boat transects that cowtails formed larger hetero‐ than monospecific groups despite the low density of whiprays. Possible benefits accrued by the cowtail were investigated in terms of predator protection. Whiprays responded earlier than cowtails to a mock predator (boat), and were most frequently the first to respond when in a mixed group. Thus, cowtails may benefit from grouping with heterospecifics by receiving earlier warning of a predator's approach. A decoy experiment using model whiprays demonstrated that cowtails were more willing to rest with models with relatively longer tails (controlled for body size). Ray tails, which are equipped with a mechanoreceptor capable of detecting predators, may constitute an important secondary means of predator detection aside from early warning. This contention is supported by the observation that stingrays mainly form resting groups when their visual ability is likely to be impaired by environmental conditions, and that tail length is negatively allometric with body size, suggesting its importance in vulnerable early life stages. If the efficacy of the mechanoreceptor increases with tail length, then cowtails may have further improved their likelihood of detecting predators by grouping with longer‐tailed heterospecifics.