Sequence effects on RNA bulge-induced helix bending and a conserved five-nucleotide bulge from the group I introns.
Bulge loops introduce bends in RNA double helices. Thus, a role for bulge loops in the tertiary folding of RNA is to orient helical elements. The location, size, and sequence of a five-nucleotide bulge are conserved in many of the self-splicing group I introns. We have used gel electrophoretic analysis of helix bending to test the hypothesis that this bulge loop is conserved to control the angle between the flanking helices. Interruption of an RNA duplex by the five-nucleotide bulge of the group I intron from Tetrahymena thermophila results in an electrophoretically retarded species, indicative of bending by the bulge. However, mutation of conserved bases in the bulge has a small effect on the retardation, suggesting that the average induced bend angle is not strongly dependent on the conserved sequence. Electrophoretic analysis of a mixture of bulged duplexes containing all five-nucleotide bulges reveals that most five-nucleotide bulge sequences induce bends that are similar to the bend induced by the conserved bulge. We have calibrated relative electrophoretic mobilities with bends of known magnitude, and characterized the distribution of bulge sequences among bend angles. Though the entire range of bend angles induced by different five-nucleotide bulges is from approximately 45 degrees to 75 degrees, most ( > 85%) five-nucleotide bulge loops induce bends between 65 degrees and 75 degrees. We have identified several of the anomalous five-nucleotide bulge sequences that induce bends of magnitude smaller than 65 degrees. They are generally, though not universally, pyrimidine-rich.


A new mechanism shapes the naïve CD8+ T cell repertoire: the selection for full diversity.
During thymic T cell differentiation, TCR repertoires are shaped by negative, positive and agonist selection. In the thymus and in the periphery, repertoires are also shaped by strong inter-clonal and intra-clonal competition to survive death by neglect. Understanding the impact of these events on the T cell repertoire requires direct evaluation of TCR expression in peripheral naïve T cells. Several studies have evaluated TCR diversity, with contradictory results. Some of these studies had intrinsic technical limitations since they used material obtained from T cell pools, preventing the direct evaluation of clonal sizes. Indeed with these approaches, identical TCRs may correspond to different cells expressing the same receptor, or to several amplicons from the same T cell. We here overcame this limitation by evaluating TCRB expression in individual naïve CD8+ T cells. Of the 2269 Tcrb sequences we obtained from 13 mice, 99% were unique. Mathematical analysis of the data showed that the average number of naïve peripheral CD8+ T cells expressing the same TCRB is 1.1 cell. Since TCRA co-expression studies could only increase repertoire diversity, these results reveal that the number of naïve T cells with unique TCRs approaches the number of naïve cells. Since thymocytes undergo multiple rounds of divisions after TCRB rearrangement and 3-5% of thymocytes survive thymic selection events the number of cells expressing the same TCRB was expected to be much higher. Thus, these results suggest a new repertoire selection mechanism, which strongly selects for full TCRB diversity.


[Progress in the application of exposomics in risk assessment of environmental chemicals].
With the increasing number of environmental chemicals, the risk assessment system based on animal toxicity testing is difficult to satisfy the current demands and needs to be reformed urgently. With the wide application of high-throughput screening technology, the advantage of exposomics in risk assessment is gradually emerging. Exposomics aims to study the health effects of mixed environmental exposure at different stages of life (especially in the early stage of life) through the dynamic monitoring of external and internal exposure levels. Exposomics is introduced into risk assessment to systematically study gene-environment interaction so as to provide a reliable basis for the construction of exposure-biomarker-disease network. This review summarized the importance and application prospect of exposomics in risk assessment of environmental chemicals in terms of the research progress, application in risk assessment and existing problems, opportunities and challenges.


Comparative proteomics analyses reveal the virB of B. melitensis affects expression of intracellular survival related proteins.
Brucella melitensis is a facultative, intracellular, pathogenic bacterium that replicates within macrophages. The type IV secretion system encoded by the virB operon (virB) is involved in Brucella intracellular survival. However, the underlying molecular mechanisms, especially the target proteins affected by the virB, remain largely unclear. In order to define the proteins affected by virB, the proteomes of wild-type and the virB mutant were compared under in vitro conditions where virB was highly activated. The differentially expressed proteins were identified by MALDI-TOF-MS. Forty-four down-regulated and eighteen up-regulated proteins which exhibited a 2-fold or greater change were identified. These proteins included those involved in amino acid transport and metabolism, lipid metabolism, energy production, cell membrane biogenesis, translation, post-translational modifications and protein turnover, as well as unknown proteins. Interestingly, several important virulence related proteins involved in intracellular survival, including VjbR, DnaK, HtrA, Omp25, and GntR, were down-regulated in the virB mutant. Transcription analysis of virB and vjbR at different growth phase showed that virB positively affect transcription of vjbR in a growth phase dependent manner. Quantitative RT-PCR showed that transcription of these genes was also affected by virB during macrophage cell infection, consistent with the observed decreased survival of the virB mutant in macrophage. These data indicated that the virB operon may control the intracellular survival of Brucella by affecting the expression of relevant proteins.


Integration of mental health into primary care in Kenya.
Integration of mental health into primary care is essential in Kenya, where there are only 75 psychiatrists for 38 million population, of whom 21 are in the universities and 28 in private practice. A partnership between the Ministry of Health, the Kenya Psychiatric Association and the World Health Organization (WHO) Collaborating Centre, Institute of Psychiatry, Kings College London was funded by Nuffield Foundation to train 3,000 of the 5,000 primary health care staff in the public health system across Kenya, using a sustainable general health system approach. The content of training was closely aligned to the generic tasks of the health workers. The training delivery was integrated into the normal national training delivery system, and accompanied by capacity building courses for district and provincial level staff to encourage the inclusion of mental health in the district and provincial annual operational plans, and to promote the coordination and supervision of mental health services in primary care by district psychiatric nurses and district public health nurses. The project trained 41 trainers, who have so far trained 1671 primary care staff, achieving a mean change in knowledge score of 42% to 77%. Qualitative observations of subsequent clinical practice have demonstrated improvements in assessment, diagnosis, management, record keeping, medicine supply, intersectoral liaison and public education. Around 200 supervisors (psychiatrists, psychiatric nurses and district public health nurses) have also been trained. The project experience may be useful for other countries also wishing to conduct similar sustainable training and supervision programmes.


Replication-Deficient Lymphocytic Choriomeningitis Virus-Vectored Vaccine Candidate for the Induction of T Cell Immunity against Mycobacterium tuberculosis.
Mycobacterium tuberculosis (Mtb) represents a major burden to global health, and refined vaccines are needed. Replication-deficient lymphocytic choriomeningitis virus (rLCMV)-based vaccine vectors against cytomegalovirus have proven safe for human use and elicited robust T cell responses in a large proportion of vaccine recipients. Here, we developed an rLCMV vaccine expressing the Mtb antigens TB10.4 and Ag85B. In mice, rLCMV elicited high frequencies of polyfunctional Mtb-specific CD8 and CD4 T cell responses. CD8 but not CD4 T cells were efficiently boosted upon vector re-vaccination. High-frequency responses were also observed in neonatally vaccinated mice, and co-administration of rLCMV with Expanded Program of Immunization (EPI) vaccines did not result in substantial reciprocal interference. Importantly, rLCMV immunization significantly reduced the lung Mtb burden upon aerosol challenge, resulting in improved lung ventilation. Protection was associated with increased CD8 T cell recruitment but reduced CD4 T cell infiltration upon Mtb challenge. When combining rLCMV with BCG vaccination in a heterologous prime-boost regimen, responses to the rLCMV-encoded Mtb antigens were further augmented, but protection was not significantly different from rLCMV or BCG vaccination alone. This work suggests that rLCMV may show utility for neonatal and/or adult vaccination efforts against pulmonary tuberculosis.


Role of circulating immune complexes and platelet aggregation in the development of experimentally induced atherosclerosis in monkeys with normal intact vas deferens and following vasectomy.
The study was undertaken in normal and vasectomized monkeys to elucidate the relationship of circulating immune complexes and platelet aggregability with experimental aortic and coronary atherosclerosis. Four groups of animals, viz. sham-vasectomized stock diet fed, vasectomized stock diet fed, sham-vasectomized atherogenic diet fed, were studied for a period of 1 year. An increased incidence of atherosclerosis was noted with high levels of circulating immune complexes in vasectomized monkeys. Platelets obtained from atherogenic diet fed monkeys had a clear tendency of increased aggregation and agglutination in the presence of ADP, epinephrine and ristocetin respectively. Vasectomy produced a significant enhancement in platelet aggregation response with ADP and epinephrine in atherogenic diet fed animals. The aortic and coronary atherosclerosis was also significantly increased in atherogenic diet fed groups both with and without vasectomy.


Prevalence of Cytauxzoon felis Infection-Carriers in Eastern Kansas Domestic Cats.
Cytauxzoon felis is a hemoprotozoal tick-transmitted pathogen of felids. Felids that survive acute disease often remain infected and serve as reservoirs for subsequent tick transmission to other susceptible felines. States adjacent to Kansas have identified C. felis-domestic cat carriers while statewide awareness and concern of cytauxzoonosis have increased. The objective of this study was to determine the prevalence of C. felis-carriers in the eastern Kansas domestic cat population using a sensitive quantitative PCR assay targeting the C. felis Cox3 mitochondrial gene. An overall C. felis infection prevalence of 25.8% was determined for asymptomatic domestic cats in eastern Kansas. Significantly more C. felis-carrier cats were identified in spring and fall, suggesting a seasonal fluctuation of survivors. Additionally, a greater percentage of feral and owned cats were positive for C. felis compared to rescue/rescinded cats. This study demonstrates that C. felis-domestic cat carriers are common among cats that spend at least a portion of time outdoors in eastern Kansas, and that more cats likely survive cytauxzoonosis than expected. Understanding the role of domestic cat carriers of C. felis is essential in developing cytauxzoonosis mitigation strategies, including recommending year-round use of acaricide products for all cats that spend any time outdoors.


Do dispositional rumination and/or mindfulness moderate the relationship between life hassles and psychological dysfunction in adolescents?
This study examined the moderating effects of dispositional rumination and mindfulness on the relationship between recent life hassles and adolescent mental health (operationalized as symptoms of depression, anxiety and stress). Data collected from a sample of 317 Australian high school students comprised an inventory of recent life hassles, measures of dispositional rumination and dispositional mindfulness and an assessment of current symptoms of depression, anxiety and stress. An increased incidence of recent life hassles was reliably associated with increased depressive symptoms, anxiety and stress. However, moderation analyses revealed that dispositional rumination exacerbated the relationship between life hassles and symptoms of depression and anxiety, whereas dispositional mindfulness attenuated the relationship between life hassles and symptoms of depression, anxiety and stress. Interventions to increase dispositional mindfulness in childhood are proposed as a method of protecting the psychological well-being of adolescents confronted by inevitable everyday life stress.


The influence of legal status on the labor market impact of immigration.
This article explores US labor market changes that would take place as a result of an amnesty that would regularize the status of undocumented workers without changing the total size of the alien workforce. The theoretical analysis suggests that the influence of legal status on market wage rates and on minimum wage enforcement is weak and that to the extent that there is an effect, it depends on particular institutional arrangements. Although data are not adequate for a definite measurement of these effects, those data that are available support this conclusion. It appears that the presence of undocumented as opposed to resident aliens can weaken union organizing efforts.


D-ribose-5-phosphate isomerase from spinach: heterologous overexpression, purification, characterization, and site-directed mutagenesis of the recombinant enzyme.
A cDNA encoding spinach chloroplastic ribose-5-phosphate isomerase (RPI) was cloned and overexpressed in Escherichia coli, and a purification scheme for the recombinant enzyme was developed. The purified recombinant RPI is a homodimer of 25-kDa subunits and shows kinetic properties similar to those of the homodimeric enzyme isolated from spinach leaves (A. C. Rutner, 1970, Biochemistry 9, 178-184). Phosphate, used as a buffer in previous studies, is a competitive inhibitor of RPI with a K(i) of 7.9 mM. D-Arabinose 5-phosphate is an effective inhibitor, while D-xylulose-5 phosphate is not, indicating that the configuration at carbon-3 contributes to substrate recognition. Although D-arabinose 5-phosphate binds to RPI, it is not isomerized, demonstrating that the configuration at carbon-2 is crucial for catalysis. Alignment of RPI sequences from diverse sources showed that only 11 charged amino acid residues of the 236-residue subunit are conserved. The possible function of four of these residues was examined by site-directed mutagenesis. D87A, K100A, and D90A mutants show greatly diminished k(cat) values (0. 0012, 0.074, and 0.38% of the wild type, respectively), while E91A retains substantial activity. Only insignificant or moderate changes in K(m) of D-ribose 5-phosphate are observed for D87A, K100A, and D90A, indicating a direct or indirect catalytic role of the targeted residues.


Intracellular and in vitro-translated 27-kDa proteins contain the 3C-like proteinase activity of the coronavirus MHV-A59.
The coronavirus mouse hepatitis virus-A59 (MHV-A59) encodes a serine-like proteinase (3C-like proteinase or 3CLpro) in ORF 1a of gene 1 between nucleotides 10,209 and 11,114. We previously have demonstrated that proteins expressed in vitro from a cDNA clone of the 3CLpro region possess proteinase activity, and that the proteinase is able to cleave substrate in trans. We sought to determine if the 27-kDa in vitro cleavage product (p27) was an active form of the 3CLpro and whether this was consistent with the 3CLpro expressed in virus-infected cells. Antibodies directed against the 3CLpro domain detected 27-kDa MHV proteins in vitro and in MHV-A59-infected cells. The 27-kDa proteins were able to cleave substrate in trans without other protein cofactors or supplemental membranes, and the p27 proteinase activity was retained after purification by immunoprecipitation and gel electrophoresis. When p27 was expressed in vitro with portions of the amino-and carboxy-terminal flanking domains (MP1 and MP2), p27 was not liberated by cls cleavage. The proteolytic activity of the 27-kDa proteins was inhibited by a variety of cysteine and serine proteinase inhibitors, and was eliminated by the cysteine proteinase inhibitor E64d. These results indicate that the 27-kDa protein is a mature proteinase in MHV-A59-infected cells, and that appropriate processing of this molecule occurs in vitro.


Inflammatory cytokine TNF-α inhibits Na(+)-glutamine cotransport in intestinal epithelial cells.
Glutamine (Gln), a preferred fuel source for enterocytes, is critical for intestinal epithelial cell integrity and barrier function. Chronic enteritis inhibits apical Na(+)-Gln cotransport. It is not known whether inflammatory cytokines that are secreted during inflammation inhibit Na(+)-Gln cotransport. Thus, this study aimed to examine whether TNF-α would affect apical Na(+)-Gln cotransport in intestinal epithelial cells. In this study, the presence of Na(+)-Gln cotransport was established by measuring Gln uptake in 10 days postconfluent IEC-6 cells grown on transwell plates. Cation, amino acid specificity, and siRNA transfection studies established that Na(+)-Gln cotransport is mediated via B(0)AT1. Immunoblotting and immunofluorescence studies established the apical membrane localization of B(0)AT1 in IEC-6 cells. Tumour necrosis factor α (TNF-α) inhibited Na(+)-Gln cotransport in a concentration- and time-dependent manner with an inhibitory concentration of 1.53 nmol·L(-1). Quantitative real-time PCR and Western blot analyses indicated that TNF-α did not alter B(0)AT1-specific transcripts or protein expression level. Kinetic studies revealed that TNF-α inhibited Na(+)-Gln cotransport by reducing the affinity of the cotransporters for Gln, and this effect was antagonized by genistein. Thus, we conclude that the TNF-α inhibition of Na(+)-Gln cotransport occurs at the post-translational level, and that the IEC-6 cell line is an excellent system to study the role of cytokines in Na(+)-Gln cotransport.


Role of the somersault rearrangement in the oxidation step for flavin monooxygenases (FMO). A comparison between FMO and conventional xenobiotic oxidation with hydroperoxides.
Model quantum mechanical calculations presented for C-4a-flavin hydroperoxide (FlHOOH) at the B3LYP/6-311+G(d,p) level suggest a new mechanism for flavoprotein monooxygenase (FMO) oxidation involving a concerted homolytic O-O bond cleavage in concert with hydroxyl radical transfer from the flavin hydroperoxide rather than an S(N)2-like displacement by the substrate on the C-4a-hydroperoxide OOH group. Homolytic O-O bond cleavage in a somersault-like rearrangement of hydroperoxide C-4a-flavinhydroperoxide (1) (FLHO-OH → FLHO···HO) produces an internally hydrogen-bonded HO(•) radical intermediate with a classical activation barrier of 27.0 kcal/mol. Model hydroperoxide 1 is used to describe the transition state for the key oxidation step in the paradigm aromatic hydroxylase, p-hydroxybenzoate hydroxylase (PHBH). A comparison of the electron distribution in the transition structures for the PHBH hydroxylation of p-hydroxybenzoic acid (ΔE(‡) = 23.0 kcal/mol) with that of oxidation of trimethylamine (ΔE(‡) = 22.3 kcal/mol) and dimethyl sulfide (ΔE‡ = 14.1 kcal/mol) also suggests a mechanism involving a somersault mechanism in concert with transfer of an HO(•) radical to the nucleophilic heteroatom center with a hydrogen transfer back to the FLH-O residue after the barrier is crossed to produce the final product, FLH-OH. In each case the hydroxylation barrier was less than that of the O-O rearrangement barrier in the absence of a substrate supporting an overall concerted process. All three transition structures bear a resemblance to the TS for the comparable hydroxylation of isobutane (ΔE(‡) = 29.2 kcal/mol) and for simple Fenton oxidation by aqueous iron(III) hydroperoxides. To our surprise the oxidation of N- and S-nucleophiles with conventional oxidants such as alkyl hydroperoxides and peracids also proceeds by HO(•) radical transfer in a manner quite similar to that for tricyclic hydroperoxide 1. Stabilization of the developing oxyradical produced by somersault rearrangement for concerted enzymatic oxidation with tricyclic hydroperoxide 1 results in a reduced overall activation barrier.


The effect of a multicomponent dual-modality exercise program targeting osteoporosis on bone health status and physical function capacity of postmenopausal women.
Exercise is important for the prevention of osteoporosis and the reduction of fracture risk because it improves muscle mass and strength, besides improving balance. We evaluated the effect of a specific exercise program on bone mass and quality and physical function capacity in postmenopausal women with low bone mineral density. Participants (N = 125) underwent a bone mass (Dual X-ray Absorptiometry), bone quality (osteosonography), and physical functional capacity assessment. Fifty-eight of them took part in an 11-month exercise program (E), that included a multicomponent (strength, aerobic capacity, balance, joint mobility) dual-modality (on ground and in the water; alternating group and home-based exercise periods) exercise regimen. The others represented a control group (C) that did not exercise. After the exercise program all participants were reevaluated. After the training program: femoral neck T-score significantly improved in E; C significantly decreased all bone quality (osteosonography) parameters, whereas E showed no differences; E significantly improved all the physical function capacity parameters, while most of them decreased or did not change in C. A specific exercise program targeting osteoporosis improves physical function capacity, reduces physiological bone loss, and maintains bone quality in low bone mineral density postmenopausal women.


Epidural perfusion cooling protection against protracted spinal cord ischemia in rabbits.
The protective effect of a modified epidural cooling technique was assessed in a rabbit spinal cord ischemia model. The epidural space around the lumbar segments with induced ischemia was continually perfused with cold (5 degrees C) isotonic saline via two communicating spinal canal openings. This procedure allowed the spinal cord to be kept deeply hypothermic (< 15 degrees C within central gray matter) during the ischemic period. The animals were subjected to either normothermic ischemia (Group A) or hypothermic ischemia (Group B). Each group contained three subgroups of animals undergoing 20, 40, or 60 minutes of aortic ligation. Their neurological outcomes were evaluated up to 48 hours postischemia, and the intergroup differences were compared. Two days postischemia, all of the animals were sacrificed by transcardial perfusion-fixation and their lumbar segments were processed for histopathological examination. In addition, in animals with 60-minute ischemia, spinal somatosensory evoked potentials were recorded during surgical intervention and again after 48 hours. In the normothermic animals, a high incidence of paraplegia was detected: in 40% after 20 minutes of ischemia, in 75% after 40 minutes, and in 100% after 60 minutes. In contrast, all of the hypothermic animals exhibited full neurological recovery even after 60 minutes of ischemia. Both electrophysiological and histological observations clearly correlated with the neurological findings. The results suggest that deep spinal cord hypothermia produced by epidural perfusion cooling provides effective protection against protracted spinal cord ischemia in rabbits.


[High expression of TCF-4 positively correlated with lung cancer progression.].
TCF-4 is an important downstream molecule of Wnt signaling pathway, but studies on the expression level and significance of TCF-4 in lung cancer were still limited. The aim of this study is to examine the expression level of TCF-4 in lung cancer tissues and cell lines, and analyze its relationship with clinicopathologic characteristics and biological behavior of lung cancers. TCF-4 expression was examined in 120 lung cancer specimens and 10 corresponding normal lung specimens using immunohistochemistry (S-P method). Immunofluorescence was used to examine the TCF-4 protein expression level and subcellular localization in lung cancer cells and HBE (human normal bronchi epithelium) cells. Expression levels of TCF-4 mRNA were examined using transcription-polymerase chain reaction (RT-PCR) in 9 lung cancer cell lines. Among 120 lung cancer specimens, 96 samples showed high expression of TCF-4 (80.0%), including 37 samples showed TCF-4 expression both in cytoplasm and nuclei (30.8%),TCF-4 were not detected in 10 corresponding normal lung specimens. TCF-4 expression level was positively correlated with TNM stages (P =0.022). The fluorescence signal of TCF-4 protein was conspicuous in lung cancer cells, primarily in cell nuclei, but extremely low in HBE. TCF-4 mRNA expression levels were incompatible in different histology types, the expression level of TCF-4 was relatively high in giant cell carcinoma cells, but low in lung adencarcinoma cells. High expression of TCF-4 positively correlated with lung cancer progression.


Randomized Controlled Trial Investigating the Effects of a Low-Glycemic Index Diet on Pregnancy Outcomes in Women at High Risk of Gestational Diabetes Mellitus: The GI Baby 3 Study.
Dietary interventions can improve pregnancy outcomes in women with gestational diabetes mellitus (GDM). We compared the effect of a low-glycemic index (GI) versus a conventional high-fiber (HF) diet on pregnancy outcomes, birth weight z score, and maternal metabolic profile in women at high risk of GDM. One hundred thirty-nine women [mean (SD) age 34.7 (0.4) years and prepregnancy BMI 25.2 (0.5) kg/m(2)] were randomly assigned to a low-GI (LGI) diet (n = 72; target GI ∼50) or a high-fiber, moderate-GI (HF) diet (n = 67; target GI ∼60) at 14-20 weeks' gestation. Diet was assessed by 3-day food records and infant body composition by air-displacement plethysmography, and pregnancy outcomes were assessed from medical records. The LGI group achieved a lower GI than the HF group [mean (SD) 50 (5) vs. 58 (5); P < 0.001]. There were no differences in glycosylated hemoglobin, fructosamine, or lipids at 36 weeks or differences in birth weight [LGI 3.4 (0.4) kg vs. HF 3.4 (0.5) kg; P = 0.514], birth weight z score [LGI 0.31 (0.90) vs. HF 0.24 (1.07); P = 0.697], ponderal index [LGI 2.71 (0.22) vs. HF 2.69 (0.23) kg/m(3); P = 0.672], birth weight centile [LGI 46.2 (25.4) vs. HF 41.8 (25.6); P = 0.330], % fat mass [LGI 10 (4) vs. HF 10 (4); P = 0.789], or incidence of GDM. In intensively monitored women at risk for GDM, a low-GI diet and a healthy diet produce similar pregnancy outcomes.


[The liver herniation through four diaphragmatic defects, simulating lobulated diaphragmatic tumor].
A 47 year-old woman, 153 cm in height and 64 kg in weight, was admitted for investigation of mass like shadow of the right lower lung field without past history of trauma and surgical treatment While magnetic resonance imaging (MRI) showed a mass 3 cm in diameter having the running vessel from the liver, computed tomography (CT) scan showed that the mass was lobulated like a tumor. We therefore performed video-assisted thoracic surgery (VATS) for diagnosis, which revealed the 4 lobulated liver tissues protruding through 4 defects of the diaphragm. The herniated parts of the liver were replaced in the abdominal cavity and diaphragmatic defects repaired by VATS. At histological features of diaphragm, thinning muscle layer was observed without any specific findings. We speculated that weakening of diaphragm was a cause of herniation. Liver herniation through multiple defects is sometimes difficult to be differentiated from diaphragmatic tumors. VATS is not only useful to diagnose it but also to repair the defects.


MacAB-TolC Contributes to the Development of Acinetobacter baumannii Biofilm at the Solid-Liquid Interface.
Acinetobacter baumannii has emerged as one of the most problematic bacterial pathogens responsible for hospital-acquired and community infections worldwide. Besides its high capacity to acquire antibiotic resistance mechanisms, it also presents high adhesion abilities on inert and living surfaces leading to biofilm development. This lifestyle confers additional protection against various treatments and allows it to persist for long periods in various hospital niches. Due to their remarkable antimicrobial tolerance, A. baumannii biofilms are difficult to control and ultimately eradicate. Further insights into the mechanism of biofilm development will help to overcome this challenge and to develop novel antibiofilm strategies. To unravel critical determinants of this sessile lifestyle, the proteomic profiles of two A. baumannii strains (ATTC17978 and SDF) grown in planktonic stationary phase or in mature solid-liquid (S-L) biofilm were compared using a semiquantitative proteomic study. Of interest, among the 69 common proteins determinants accumulated in the two strains at the S-L interface, we sorted out the MacAB-TolC system. This tripartite efflux pump played a role in A. baumannii biofilm formation as demonstrated by using ΔmacAB-tolC deletion mutant. Complementary approaches allowed us to get an overview of the impact of macAB-tolC deletion in A. baumannii physiology. Indeed, this efflux pump appeared to be involved in the envelope stress response occurring in mature biofilm. It contributes to maintain wild type (WT) membrane rigidity and provides tolerance to high osmolarity conditions. In addition, this system is probably involved in the maintenance of iron and sulfur homeostasis. MacAB-TolC might help this pathogen face and adapt to deleterious conditions occurring in mature biofilms. Increasing our knowledge of A. baumannii biofilm formation will undoubtedly help us develop new therapeutic strategies to tackle this emerging threat to human health.


Regulation of vascular smooth muscle cell calcification by syndecan-4/FGF-2/PKCα signalling and cross-talk with TGFβ.
Vascular calcification is a major cause of morbidity and mortality. Fibroblast growth factor-2 (FGF-2) plays an instructive role in osteogenesis and bone development, but its role in vascular calcification was unknown. Therefore, we investigated the involvement of FGF-2 in vascular calcification and determined the mechanism by which it regulates this process. We demonstrate that FGF-2 expression is increased in vascular smooth muscle cells (VSMCs) induced to deposit a mineralized matrix by incubation with β-glycerophosphate. FGF-2 is also localized to sites of calcification within human atherosclerotic plaques. The expression of syndecan-4, a heparan sulfate proteoglycan which regulates FGF-2 signalling, is also increased in mineralizing VSMCs and co-localizes with FGF-2 in human calcified atherosclerotic plaques. Exogenous FGF-2 inhibits VSMC mineralization, and this inhibition is reduced when syndecan-4 expression is knocked-down using siRNA. Biochemical inhibition of FGFR signalling using a pan FGFR inhibitor (BGJ398) or knocking-down syndecan-4 expression in VSMCs using siRNA increases VSMC mineralization. These increases are prevented by inhibiting transforming growth factor-β (TGFβ) signalling with SB431542, suggesting cross-talk between FGF-2 and TGFβ signalling is crucial for the regulation of VSMC mineralization. Syndecan-4 can also regulate FGF-2 signalling directly via protein kinase Cα (PKCα) activation. Biochemical inhibition of PKCα activity using Gö6976, or siRNA-mediated suppression of PKCα expression increases VSMC mineralization; this increase is also prevented with SB431542. Finally, the ability of FGF-2 to inhibit VSMC mineralization is reduced when PKCα expression is knocked-down. This is the first demonstration that syndecan-4 promotes FGF-2 signalling, and in turn, suppresses VSMC mineralization by down-regulating TGFβ signalling. Our discoveries that FGF-2 and syndecan-4 expression is increased in mineralizing VSMCs and that PKCα regulates FGF-2 and TGFβ signalling in VSMCs suggests that the syndecan-4/FGF-2/TGFβ signalling axis could represent a new therapeutic target for vascular calcification.


Interaction of multiwalled carbon nanotubes with supported lipid bilayers and vesicles as model biological membranes.
The influence of solution chemistry on the kinetics and reversibility of the deposition of multiwalled carbon nanotubes (MWNTs) on model biological membranes was investigated using a quartz crystal microbalance with dissipation monitoring (QCM-D). Supported lipid bilayers (SLBs) comprised of zwitterionic 1,2-dioleoyl-sn-glyero-3-phosphocholine (DOPC), as well as DOPC vesicles, were used as model cell membranes. Under neutral pH conditions, the deposition kinetics of MWNTs on SLBs increased with increasing electrolyte (NaCl and CaCl2) concentrations. In the presence of NaCl, favorable deposition was not achieved even at a concentration of 1 M, which is attributed to the presence of strong repulsive hydration forces due to the highly hydrophilic headgroups of SLBs. Conversely, favorable deposition was observed at CaCl2 concentrations above 0.5 mM when the charge of SLBs was reversed from negative to positive through the binding of Ca(2+) cations to the exposed phosphate headgroups. Favorable nanotube deposition was also observed at pH 2, at which the DOPC SLBs exhibited positive surface charge, since the isoelectric point of DOPC is ca. 4. When MWNTs on SLBs were rinsed with low ionic strength solutions at pH 7.3, only ca. 20% of deposited nanotubes were released, indicating that nanotube deposition was mostly irreversible. The deposition of MWNTs on DOPC vesicles under favorable deposition conditions did not result in any detectable leakage of solution from the vesicles, indicating that MWNTs did not severely disrupt the DOPC bilayers upon attachment.


[Construction, expression and identification of recombinant plasmid encoding bifunctional protein sflk1-IFN-gamma].
To construct, express and identify the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma encoding bifunctional protein sflk1-IFN-gamma (soluble fetal liver kinase 1 and interferon-gamma). sflk1 and IFN-gamma gene fragments were cloned by RT-PCR, and then inserted into pcDNA3.1(+) plasmid between BamHI-EcoRI and XhoI-XbaI restriction sites to form the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma. The recombinant sflk1-IFN-gamma transiently expressed in COS-7 cells was detected by ELISA and Western blotting. Bioactivities of sflk1-IFN-gamma fusion protein were identified by proliferation inhibition assay with H5V cells and NK activity assay. pcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in COS-7 cells. Concentrations of sflk1 and IFN-gamma in culture supernatants of pcDNA3.1(+)/sflk1-IFN-gamma transfected COS-7 cells were (20.85+/-2.48) ng/ml and (1.08+/-0.09) ng/ml, respectively. Western blotting showed that the molecular weight of sflk1-IFN-gamma fusion protein was about 130 kDa, while that of sflk1 was 115 kDa. The supernatants of transfected cells significantly inhibited the proliferation of H5V cells stimulated by mouse VEGF 164 and enhanced the NK activity of splenocytes, demonstrating that sflk1-IFN-gamma fusion protein possessed the bioactivities of both sflk1 and IFN-gamma. The constructed plasmid pcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in eukaryotes. The expressed sflk1-IFN-gamma fusion protein has the biological activities of both sflk1 and IFN-gamma.


Microbial thiamin metabolism in the rumen simulating fermenter (RUSITEC): the effect of acidogenic conditions, a high sulfur level and added thiamin.
The effects of acidogenic conditions, a high S level and the addition of thiamin on the rumen microbial metabolism of thiamin were investigated in vitro in a semi-continuous fermenter (RUSITEC), using a factorial design. Acidogenic conditions were obtained by simultaneously increasing the starch: cellulose ratio and the amount of solid substrate fed, and by decreasing the buffering capacity of the liquid phase of the fermenter. S in the form of sulfate was supplied at two levels, one corresponding to a control amount of S (2 g/kg dietary DM), the second to an excess (5 g/kg DM) which is sufficient to trigger cerebrocortical necrosis (CCN) when used in vivo. Acidogenic conditions decreased the pH of the fermenters, CH4 production and cellulose digestibility, increased the short-chain fatty acid production, but had no effect on thiamin production. The high S level enhanced the production of sulfide considerably, had no effect ont he microbial metabolism of energy and N, and decreased thiamin production (326 v. 266 nmol/d). The added thiamin was rapidly converted into phosphorylated compounds which largely decreased the apparent synthesis of this vitamin by the rumen microflora. The total thiamin flow was increased by added thiamin. In no case was thiaminase activity in the fermenter liquid phase significantly modified. The high level of S induced only a limited decrease of total thiamin flow. Consequently, it is unlikely that the investigated factors could be considered to be high risk factors for the thiamin-dependent CCN.


Lumbosacral hemivertebrae resection by combined approach: medium- and long-term follow-up.
Retrospective review of patients records with clinical and radiographic assessment. To evaluate the long-term result of lumbosacral hemivertebrae resection using a combined approach. Lumbosacral hemivertebrae resection by a combined posterior and anterior approach have been previously described, but only one study is concerning such a large series of hemivertebrae. From 1980 to 2001, a consecutive series of 17 congenital scolioses due to lumbosacral hemivertebrae were managed by hemivertebrae resection using a combined approach and short anterior and posterior convex fusion. The mean age at surgery was 4.9 years. The mean follow-up period was 7.6 years. One postoperative complication was encountered: a patient had motor deficit and recovery was incomplete. The mean segmental scoliosis curve was 27.5 degrees before surgery, 10.7 degrees after surgery, and 8.9 degrees at the latest follow-up assessment. This represents a mean improvement of 61.1% after surgery and of 67.6% at the latest follow-up. The total main scoliosis curve improved from 28.4 degrees to 12.7 degrees and 11.4 degrees , respectively. It represents a mean improvement of 55.3% and 59.9%, respectively. The mean final lordosis was within normal values. This procedure offers a persistent correction with a short-segment fusion. The early surgery is able to avert severe local deformities and prevent secondary structural deformities in order to avoid extensive fusions.


Development of multiplex reverse transcription-polymerase chain reaction for differentiation of strains of infectious bursal disease virus and primary screening of the virus in Thailand.
A new set of primers (400 base pairs partial of VP2) was designed and used for the infectious bursal disease virus (IBDV) screening test. Using this new primer set, the enzymes MboI and BstNI were unable to differentiate the field and vaccine strains. As a result, a new simple, cheap, and appropriate tool for strain differentiation is required. The objective of this study was to develop the appropriate restriction fragment length polymorphism (RFLP) and multiplex reverse transcription-polymerase chain reaction (RT-PCR) for the differentiation of classic IBDV (cIBDV) strains and very virulent IBDV (vvIBDV) strains in Thailand. Ninety seven bursa of Fabricius from 16 farms were collected from farms in the eastern and central regions of Thailand. RT-PCR screening showed that 82 samples were positive for IBDV and 15 samples were negative. Then, selected samples were sequenced from each farm with a positive test. The sequencing results showed that samples from six of the farms were vvIBDV and samples from the other six farms were cIBDV. Although the whole genome sequencing was incomplete, both the sequencing results of segment A and segment B showed high similarity between cIBDV and vvIBDV. Restriction enzyme cutting site and primers for multiplex RT-PCR were hard to design. An RT-PCR-RFLP method was developed, but it failed to differentiate IBDV strains. However, the multiplex RT-PCR was able to differentiate cIBDV from vvIBDV. Four primers were used in the multiplex RT-PCR. These four primers were used together in one reaction at an annealing temperature of 45°C. Therefore, multiplex RT-PCR is a less complicated, cheaper, and less time-consuming method for the differentiation of cIBDV and vvIBDV strains.


Pacing strategy and the occurrence of fatigue in 4000-m cycling time trials.
The present study was designed to examine the role of central and peripheral fatigue on 4000-m cycling time trial performance by comparing changes in power output and integrated electromyography (iEMG) in differently paced maximal efforts. Eight well-trained men performed three randomly ordered time trials with different pacing strategies, in which the first 2000 m were manipulated to evoke an increasing, even, and decreasing power output profile (SUB, EVEN, and SUPRA, respectively). Subjects were instructed to finish the last 2000 m of all trials in the shortest time possible. iEMG of the rectus femoris (RF), vastus lateralis (VL), and biceps femoris (BF) muscle, mechanical power output, and gas exchange variables were measured. Anaerobic and aerobic contributions to mechanical power output were calculated from gas exchange data. The increase in mechanical power output during the SUB time trials was always associated with an increase in iEMG in all muscles. A decrease in mechanical power output near the end of the time trials was also marked by an increase in iEMG for all muscles, except for the RF. Comparing the last 2000-m interval with the first, aerobic power output increased for all strategies. Anaerobic power output increased in SUB and decreased in EVEN and SUPRA. The relationship between iEMG and mechanical power output pattern was consistent with peripheral fatigue rather than central downregulation of mechanical power output. Specifically, anaerobic energy resources seem to be important in regulating pacing strategy.


Cutaneous Filariasis in Free-Ranging Rothschild's Giraffes (Giraffa camelopardalis rothschildi) in Uganda.
Across Africa, wild giraffes suffer from a variety of skin disorders of mostly unknown etiology. With their populations already threatened from anthropogenic factors, it is important to understand infectious disease risks to giraffes. Here we describe filarid parasites and a portion of their genetic sequence associated with skin disease in Rothschild's giraffes (Giraffa camelopardalis rothschildi) in Uganda.


[Thromboembolic complications in day case surgery].
Prevention of venous thrombosis in surgical hospital patients hasgained attention due to the fact that in spite of therapy, deep vein thrombosis and pulmonary embolism can cause long-term impediment and even mortality. In day case surgery, the threat of venous thrombosis has been considered low and few studies on the subject are available. On the other hand, increasingly diseased and aged patients are operated on day case surgery basis and more demanding procedures are performed for patients being discharged from the hospital during the same day. In addition, more information is constantly obtained concerning the significance of risk factors.


Vasorelaxant effects of and receptors for atrial natriuretic peptides in the mesenteric artery and aorta of the rat.
Vasorelaxant effects of different atrial natriuretic peptides (ANP) were measured on rat aortic strips and mesenteric artery rings. These results were compared with the potency of the same peptides to displace 125I-labelled ANP (101-126) on membrane preparations of aorta and of mesenteric vascular bed. In aortic strips and mesenteric artery rings precontracted with phenylephrine (3 X 10(-8) and 10(-6) M, respectively), the order of potency of ANP was as follows: ANP (99-126) greater than ANP (101-126) greater than ANP (103-126) = ANP (103-125) much greater than ANP (103-123). In the displacement binding assays, the order of potency of ANP peptides was similar to that of the relaxation experiments: ANP (99-126) = ANP (101-126) greater than ANP (103-126) = ANP (103-125) much greater than ANP (103-123). When the vessels were precontracted by a smaller concentration of phenylephrine (10(-7) M in mesenteric artery and 10(-8) M in aorta), the IC50 of ANP (101-126) was significantly lower than when the higher concentration of phenylephrine was used. These results show that ANP receptors in the mesenteric artery and in the aorta have similar structural requirements, according to the order of potency of different length ANP, both for binding and myotropic responses.


Measuring Process Capability in a Hospital by Using Lean Six Sigma Tools-A Case Study in Bangladesh.
Bangladesh's government had been constantly conscious of its health-care system. As a result, the health-care system had seen a significant rise in quality in the past few years. The Lean Six Sigma concept is used to boost up the productivity as well as the quality of products or services. The purpose of this article was to measure the process capability in the health-care system by lean tools. In this study, we selected the dialysis unit as our investigation area where we applied the Six Sigma DMAIC (Define, Measure, Analysis, Improve, and Control) model which had been structured step by step, respectively. Furthermore, different tools of Total Quality Management, Statistical Quality Control and Lean manufacturing like SIPOC (Supplier, Input, Process, Output, and Customer) diagram, P Control chart, Fishbone diagram, and Pareto analysis had been performed in different phases of the DMAIC model. Measuring process capability using the DMAIC model helped to identify the problems associated with the dialysis unit and also recommendations were developed based on investigation and analysis. These recommendations will help the hospital management to overcome all problems and increase service rate and patient safety in the dialysis unit.


Gas-uptake pharmacokinetics and metabolism of 2-chloro-1,1,1,2-tetrafluoroethane (HCFC-124) in the rat, mouse, and hamster.
Gas-uptake pharmacokinetics and metabolism of the chlorofluorocarbon replacement 2-chloro-1,1,1,2-tetrafluoroethane (HCFC-124) were investigated in rats, mice, and hamsters. Species differences in the rate of uptake of HCFC-124 and urinary excretion of trifluoroacetic acid were observed. In rats and mice, the uptake of HCFC-124 was described by both saturable and first-order components, whereas in the hamster only first-order uptake was observed. The in vivo metabolic rate constants obtained from computer simulation of the gas-uptake data were: for rats-KM = 1.2 mg liter-1 (8.79 mmol liter-1, Vmaxc = 0.35 +/- 0.01 mg kg-1 hr-1 (2.56 +/- 0.01 mmol kg-1 hr-1), and kfc = 1.25 +/- 0.01 hr-1 kg231; for mice-KM = 1.2 mg liter-1 (8.79 mmol liter-1), Vmaxc = 1.78 +/- 0.01 mg kg-1 hr-1 (13.0 +/- 0.007 mmol kg-1 hr-1), and kfc = 4.08 +/- 0.01 hr-1 kg-1; and for hamsters-kfc = 1.47 +/- 0.02 hr-1 kg-1. The production and excretion of trifluoroacetic acid, the major urinary metabolite of HCFC-124, were also simulated in rats and mice, but not in hamsters, by the physiologically based pharmacokinetic model when the in vivo metabolic rate constants obtained in the gas-uptake simulation studies were used. The blood:air partition coefficient of HCFC-124 in the hamster was lower than in the rat or mouse. A low blood:air partition coefficient may limit the pulmonary uptake of volatile chemicals.(ABSTRACT TRUNCATED AT 250 WORDS)


Journey Toward Improved Long-Term Outcomes After Norwood-Sano Procedure: Focus on the Aortic Arch Reconstruction.
The disadvantage of right ventricle-to-pulmonary artery (RV-PA) shunt is the need for more unplanned interventions to address stenosis in the shunt or branch pulmonary arteries, as compared to the modified Blalock-Taussig shunt group. Ring-enforced RV-PA PTFE conduit and dunk technique minimized these complications and right ventricle (RV) damage. Aortic arch obstruction increases afterload and leads to ventricular dysfunction and tricuspid regurgitation; therefore, most surgeons prefer to use homograft, autologous pericardium, or bovine pericardium to reconstruct the neoaorta. Artificial materials decrease the elastic properties, increase wall stiffness, and decrease the distensibility of the aorta; and as a result, RV function gradually deteriorates. This inelastic reconstructed aorta may be one of the reasons why long-term outcomes after the Fontan procedure are worse in hypoplastic left heart syndrome (HLHS) patients, in comparison to non-HLHS. Reconstruction of the neoaorta without any patch materials, or at least techniques that largely minimize the use of non-autologous materials, will offer a further refinement of our ability to optimize ventriculoarterial coupling and thereby long-term RV function.


A pilot study of the effect of the 5-HT-uptake inhibitor, zimelidine, on phobic anxiety.
A new antidepressant, zimelidine, which is a selective inhibitor of 5-HT-uptake, was tested in an open study of 13 patients, in each of whom the principal clinical diagnosis was phobic neurosis. The dose varied from 200 to 300 mg daily. After 6 weeks of treatment, 6 patients dropped out of the study as they were not appreciably improved, but 7 patients definitely improved and completed a treatment course of at least 12 weeks. In these cases it was possible to observe subjective relief of symptoms, improved social function, increased working capacity and a reduced need for anxiolytics. The effect appeared similar to the previously described effects of clomipramine treatment. The incidence of side effects was low and those that occurred were mild. Tolerance was assessed as very good by the great majority of patients. The study suggests that zimelidine has a favourable effect on mild, moderate, and even in some severe phobic conditions. Some comparisons with MAO-inhibitors were made. The results are sufficiently encouraging to suggest a controlled study.


Renal Allograft Torsion: US and CT Imaging Findings of a Rare Posttransplant Complication.
Vascular torsion is a rare renal transplant complication which requires prompt diagnosis and surgery to salvage allograft function. We report here a case of renal allograft torsion with interesting imaging findings on unenhanced CT and color Doppler ultrasound. A 60-year-old woman with a history of pancreas and kidney transplant presented to the emergency room with nausea, vomiting, abdominal pain, and minimal urine output. Unenhanced CT of the abdomen demonstrated an enlarged and malrotated renal allograft with moderate hydronephrosis. Color Doppler ultrasound demonstrated lack of vascularity within the allograft. The patient was taken urgently to the operating room where the renal allograft was found twisted 360 degrees around the vascular pedicle. After the allograft was detorsed, the color of the kidney returned and the Doppler signals for arterial flow improved. Intraoperative biopsy showed no evidence of infarct or acute cellular rejection. The detorsed kidney was surgically fixed in position in its upper and lower poles. Follow-up ultrasound 1 day later demonstrated normal blood flow to the renal allograft and the serum level of creatinine returned to normal.


Appendicitis epiploicae: An unusual cause of acute abdomen in children.
Appendicitis epiploicae is described as a rare entity in an 8-year-old boy presented with the features of acute abdomen simulating acute appendicitis. Surgical exploration revealed a torsion of appendices epiploicae of the cecum. The excision of infarcted epiploicae with seromuscular inversion resulted in satisfactory recovery. Authors describe this uncommon cause of pediatric acute abdomen along with the review of pertinent literature.


Testing a core emotion-regulation prediction: does early attentional persistence moderate the effect of infant negative emotionality on later development?
To test the hypothesis that early attentional persistence will moderate the effect of infant negative emotionality on social competence, problem behavior, and school readiness at age 3, data collected as part of the NICHD Study of Early Child Care were subject to structural equation modeling analyses (N = 1,038). Consistent with Eisenberg et al.'s data on older children, high levels of negative emotionality were associated with low levels of social competence only when attentional persistence was poor. No such moderating effects of attentional persistence emerged in the case of behavior problems. And in the case of school readiness, findings indicated that high levels of negative emotionality predicted high levels of school readiness when attentional persistence was high, a result opposite to that found with respect to the prediction of social competence.


Rifaximin alters intestinal bacteria and prevents stress-induced gut inflammation and visceral hyperalgesia in rats.
Rifaximin is used to treat patients with functional gastrointestinal disorders, but little is known about its therapeutic mechanism. We propose that rifaximin modulates the ileal bacterial community, reduces subclinical inflammation of the intestinal mucosa, and improves gut barrier function to reduce visceral hypersensitivity. We induced visceral hyperalgesia in rats, via chronic water avoidance or repeat restraint stressors, and investigated whether rifaximin altered the gut microbiota, prevented intestinal inflammation, and improved gut barrier function. Quantitative polymerase chain reaction (PCR) and 454 pyrosequencing were used to analyze bacterial 16S ribosomal RNA in ileal contents from the rats. Reverse transcription, immunoblot, and histologic analyses were used to evaluate levels of cytokines, the tight junction protein occludin, and mucosal inflammation, respectively. Intestinal permeability and rectal sensitivity were measured. Water avoidance and repeat restraint stress each led to visceral hyperalgesia, accompanied by mucosal inflammation and impaired mucosal barrier function. Oral rifaximin altered the composition of bacterial communities in the ileum (Lactobacillus species became the most abundant) and prevented mucosal inflammation, impairment to intestinal barrier function, and visceral hyperalgesia in response to chronic stress. Neomycin also changed the composition of the ileal bacterial community (Proteobacteria became the most abundant species). Neomycin did not prevent intestinal inflammation or induction of visceral hyperalgesia induced by water avoidance stress. Rifaximin alters the bacterial population in the ileum of rats, leading to a relative abundance of Lactobacillus. These changes prevent intestinal abnormalities and visceral hyperalgesia in response to chronic psychological stress.


Transcatheter electrical AV junction ablation: predictors of success.
The initial and long-term results of transcatheter electrical ablation in 29 patients with drug-refractory supraventricular tachyarrhythmias were analyzed. Ablation was immediately successful (defined as induction of chronic complete heart block) in 25 patients (86.2%). Among the group in whom ablation was unsuccessful, there were more patients with ectopic atrial tachycardia and a higher incidence of narrow QRS escape rhythm following the initial ablation. A His amplitude equal to or greater than 0.3 mV was correlated with success. Complications of ablation included deep venous thrombosis and ventricular arrhythmias. Post-ablation stress testing was superior to ambulatory monitoring in identifying early return of atrioventricular conduction.


The work hardening program and subsequent return to work of a client with low back pain.
Billions of dollars are spent annually for the costs associated with workers' compensation cases. Chronic low back injuries have been traditionally the most costly cases. In addition to the physical problems, psychosocial issues emerge which confound conventional treatment and prolong resolution. Work hardening programs were developed to address the many issues of the occupationally injured chronic patient. This case study examines the treatment of a 54-year-old male who sustained an injury to his lumbar spine during employment as a boiler mechanic and subsequently had been out of work for 8 months. The article describes the interdisciplinary work hardening treatment (5-week duration) and the progressive resistive work circuit which preceded the client's return to work. It is concluded that work hardening programs can be cost-effective for insurers, as in this case, saving an estimated $44,000. An aggressive return to work program should also be considered in earlier stages of injury as a way of preventing the development of chronic disability for which the human costs are immeasurable.


Nanocomposite ZnO-SnO2 Nanofibers Synthesized by Electrospinning Method.
We report the characterization of mixed oxides nanocomposite nanofibers of (1 - x) ZnO-(x)SnO2 (x ≤ 0.45) synthesized by electrospinning technique. The diameter of calcined nanofibers depends on Sn content. Other phases like SnO, ZnSnO3, and Zn2SnO4 were absent. Photoluminescence studies show that there is a change in the blue/violet luminescence confirming the presence of Sn in Zn-rich composition. Present study shows that the crystalline nanocomposite nanofibers with stoichiometry of (1 - x)ZnO-(x)SnO2 (x ≤ 0.45) stabilize after the calcination and possess some morphological and optical properties that strongly depend on Sn content.


Telehealth Utilization in Oral Medicine and Oral and Maxillofacial Surgery.
Objectives: The objectives of this retrospective study were to analyze telehealth utilization for two specialty care practices: oral medicine (OM) and oral and maxillofacial surgery (OMFS) during the first 2 years of the pandemic, its impact as a new treatment modality and on participating providers, as well as identify the type of patient visit that most readily adopted telehealth. Methods: Retrospective study of patients who sought specialty services, OM and OMFS, at an outpatient clinic in a university health system setting between March 1, 2019, and February 28, 2022. Source data were obtained from Epic, an electronic medical record application. Data were graphed using Tableau and Microsoft Excel software. Statistical analysis was performed utilizing chi-squared test and analysis of variance (ANOVA). Results: OMFS utilized telehealth 12% of the time, and OM 8% of the time. The majority (87%) of telehealth visits were for return patients (RPs). Compared with the first year of the pandemic, there was a decrease in the number of telehealth visits in the second year (p = 0.0001). As of August 2022, new patient (NP) telehealth encounters have largely returned to prepandemic levels (0-1.5%), whereas RP telehealth visits remained at an average level of 11.4% (9.4-12.4%). Surveyed providers consider telehealth as an effective complement to in-person care and will continue its use (4.2/5 Likert scale). Conclusions: Telehealth has become a viable pathway of care for OM and OMFS who previously did not utilize the remote platform to deliver healthcare. As a new treatment modality, telehealth is perceived as impactful in increasing access to specialty care by participating providers. NP visits are now almost completely in person, but telehealth continues for RPs. Ongoing demand for telehealth highlights urgency to develop appropriate standards and effective remote diagnostic/monitoring tools to maximize telehealth's capability to leverage finite health care resources and increase access to specialty care.


In vitro and in vivo effects of the carbon monoxide-releasing molecule, CORM-3, in the xenogeneic pig-to-primate context.
Carbon monoxide (CO) interferes with inflammatory and apoptotic processes associated with ischemia-reperfusion injury and graft rejection. Here, the in vitro effects of carbon monoxide releasing molecule-3 (CORM-3), a novel water-soluble carbonyl CO carrier, have been investigated on porcine aortic endothelial cells (PAEC) and primate peripheral blood mononuclear cells (PBMC). Furthermore, the pharmacodynamics and pharmacotolerance of CORM-3 after administration of single and multiple doses in the primate have been assessed in view of its potential application in pig-to-primate xenotransplantation models. For in vitro studies, PAEC and primate PBMC were exposed for 24, 48 and 72 h to CORM-3 (20 to 1000 microm) and viability was measured using an MTS assay. PAEC and primate PBMC proliferation after exposure to CORM-3 was assessed by CFSE labelling. Proliferation of primate PBMC against irradiated pig lymphocytes was also assessed. Tumor necrosis factor alpha (TNF-alpha) production and Caspase-3 and -7 activity in Concanavalin A (conA)-stimulated primate PBMC were measured following treatment with CORM-3. In vivo, CORM-3 was administered i.v. to cynomolgus monkeys at 4 mg/kg, as single or multiple doses for up to 30 days. The effect of CORM-3 was evaluated by the assessment of production of TNF-alpha and interleukin 1beta following PBMC stimulation with LPS by species-specific ELISA. Complete hematologic and biochemical analyses were routinely performed in treated primates. At concentrations <500 microm, CORM-3 did not alter the viability of PAEC or primate PBMC cultures in vitro, nor did it induce significant levels of apoptosis or necrosis. Interestingly, at concentrations of 300 and 500 microm, significant PAEC proliferation was observed, whilst concentrations > or =50 microm inhibited conA-activated primate lymphocyte proliferation (IC(50) of 345.8 +/- 51.9 microm) and the primate xenogeneic response against pig PBMC. Such responses were demonstrated to be CO-dependent. In addition, CORM-3 significantly inhibited caspase-3 and -7 activity at concentrations between 200 and 500 microm and caused a significant reduction in TNF-alpha production (IC(50) 332.8 +/- 33.9 microm). In vivo, following the administration of multiple doses, TNF-alpha production was significantly reduced in comparison to pre-treatment responses, with decreased levels maintained throughout the study. Moreover, a slight and transient increase in transaminases and bilirubin was observed in animals exposed to multiple doses of CORM-3. These studies suggest that CORM-3 has anti-inflammatory and immunomodulatory properties in primates that may result in clinical benefit to allo- and xenografted organs.


Diagnostic and prognostic significance of chromosome abnormalities in childhood acute lymphoblastic leukemia.
Current intensive chemotherapies cure about 70% of the children with ALL. On the other hand a significant number of the children are not cured despite intensive treatment. At the same time some highly curable patients are treated too intensively and suffer from unnecessary side effects of the chemo- and radiotherapy applied. In order to further improve the therapeutic results in this disease, we have to distinguish between the cases with a better and a worse prognosis. The initial karyotype (both numerical and structural chromosome abnormalities) proved to be one of the most reliable prognostic parameters, leading to the suggestion of developing genotype-specific therapies. Although the prognosis in patients with pseudodiploid karyotype is usually unfavorable, a significantly better prognosis can be observed in those with more than 50 chromosomes. Because the latter patients can achieve remission on a metabolite-based therapy, the toxic effects of more aggressive chemotherapy with anthracyclines and genotoxic agents can be avoided; thus, the reliable and accurate identification of patients with > 50 chromosomes is of particular importance. For this purpose three methods: chromosome analysis, DNA flow cytometry, and fluorescence in situ hybridization can be used. In 1993 it was decided to develop a comprehensive nationwide project in order to perform the initial genetic analysis of all ALL children diagnosed in the hematological/oncological centers of Hungary. Here the data obtained on 187 ALL patients diagnosed in the period from 1993 to 1995 are presented. In about 75% of patients (in 140 of 187) chromosome analysis was performed, in 78 cases (55.7%) successfully. The proportion of patients with abnormal karyotype was 36 of 78 (46.1%), and hyperdiploidy with more than 50 chromosomes was detected in 13 of 78 (16.6%) children. The lower ratio of hyperdiploid cases in our patients as compared to the data in the literature may be due to technical difficulties and the small number of patients studied, but it may reflect real geographic characteristics. Using flow cytometry, seven of 31 patients investigated (22.5%) proved to be hyperdiploid with a DNA index above 1.16. A higher ratio of hyperdiploid patients in this study calls attention to the significance of simultaneous application of the two methods. Taken together, 16 of 80 (20.0%) successfully studied patients proved to be hyperdiploid (> 50 chromosomes and/or DNA index above 1.16). The pattern of chromosome involvement in our study determined by chromosome analysis and/or FISH technique proved also to be different from the data of large international series. In addition to trisomies of chromosomes 4, 6, 10, 14, 17, 18, 21, and X, which are known to be the most frequently involved chromosomes, trisomies of chromosomes 3, 8, 11, and 13 were also observed with a high frequency. Comparison of survival curves of various cytogenetic subgroups showed a significant difference between diploid-pseudodiploid and diploid-hyperdiploid A (with 47-50 chromosomes) subgroups. No favorable prognosis of hyperdiploid patients (> 50 chromosomes) could be proved. Because of the small number of patients studied, prognostic differences of cytogenetic subgroups need further confirmation. The clinical and genetic differences observed, however, call attention to the necessity for further genetic studies of ALL patients in Hungary, because these differences may reflect real geographic characteristics and may be related to different environmental mutagen/carcinogen effects of the given geographic area. It is essential to determine whether or not these differences really exist and if they do to reveal the causes leading to these differences. In our view this is one of the routes by which the therapeutic results in childhood ALL can be further improved simultaneously with the avoidance of early and late toxicity of chemotherapy.


Mechanism of a Synergistic Effect of Kinetin on Auxin-induced Ethylene Production: Suppression of Auxin Conjugation.
In hypocotyl segments of mung bean (Phaseolus mungo L.) seedlings, exogenously supplied indoleacetic acid was rapidly conjugated mainly into indoleacetylaspartic acid, which was inactive in inducing ethylene production. Kinetin is known to stimulate indoleacetic acid-induced ethylene production. The mechanism of kinetin action on indoleacetic acid-induced ethylene production by hypocotyl segments of mung bean seedlings was studied in relation to indoleacetic acid uptake and indoleacetic acid metabolism. Kinetin enhanced indoleacetic acid uptake during the initial 2-hour incubation and markedly suppressed the conversion of indoleacetic acid to indoleacetic acid conjugates throughout the whole 7-hour incubation. As a result, there was more free indoleacetic acid and less conjugated indoleacetic acid in the segments treated with kinetin than in those receiving no kinetin. A close relationship was demonstrated between the rate of ethylene production and the level of free indoleacetic acid, which was regulated by kinetin.


Serum-free adapted Drosophila S2R+ line is amenable to RNA interference.
We have previously adapted a select number of Drosophila cell lines to grow in serum-free media supplemented with fly extract. This condition is arguably more representative of a native growth environment. Here, we validated that the fly extract adapted line, S2R+ (FEx 2.5%) is amenable to RNAi. RNAi against Rho1 in both S2R+ and S2R+ (FEx 2.5%) produced phenotypes similar to ones previously described in Drosophila S2 cells.


Pressure probe technique for measuring water relations of cells in higher plants.
A new method is described for continuously measuring cell turgor pressure (P), hydraulic conductivity (L(p)), and volumetric elastic modulus (epsilon) in higher plant cells, using a pressure probe. This technique permits volume changes, DeltaV, and turgor pressure changes, DeltaP, to be determined with an accuracy of 10(-5) to 10(-6) mul and 3 to 5.10(-2) bar, respectively.The main principle of the new method is the same as the pressure probe developed by Zimmermann and Steudle in which pressure is transmitted to a pressure transducer by means of an oil-filled capillary introduced into the cell. In order to use the pressure probe for small tissue cells, the effective compressible volume of the apparatus has to be sufficiently small in comparison to the volume of the cell itself. This is achieved by accurately fixing the oil/cell sap boundary in the very tip of the microcapillary by means of an electronic feedback mechanism, so that the effective volume of the apparatus is reduced to about 2 to 10% of the cell volume. In this way also, errors arising from compressibility of the apparatus and temperature fluctuations can be excluded.Measurements on tissues cells of Capsicum annuum fruits yield epsilon values of 2 to 25 bar. Furthermore, epsilon can be shown to be a function of both cell turgor pressure and cell volume; epsilon increases with increasing turgor pressure and is higher in larger cells.


Secreted or nonsecreted forms of acidic fibroblast growth factor produced by transfected epithelial cells influence cell morphology, motility, and invasive potential.
Addition of exogenous acidic fibroblast growth factor (aFGF) to NBT-II epithelial carcinoma cells results in fibroblastic transformation and cell motility. We have generated aFGF-producing NBT-II cells by transfection with recombinant expression vectors containing human aFGF cDNA, or the human aFGF cDNA coupled to a signal peptide (SP) sequence. The effects of the nonsecreted and the secreted 16-kDa growth factor on the morphology, motility, and cell invasive potential (gelatinase activity) were compared. aFGF coupled to a SP was actively secreted out of the producing cells. The secretion of aFGF was not necessary for induction of gelatinase activity, as this was observed in NBT-II cells producing aFGF with or without SP. Production of aFGF, whether secreted or not secreted, resulted in increased in vitro motility of most isolated clones; however, there was no correlation between aFGF level and motility rate. The data suggest that expression of aFGF in NBT-II cells induces metastatic potential through an autocrine or intracrine mechanism.


Frailty and Nutrition: What We Have Learned from Research and Clinical Practice on the Mini Nutritional Assessment.
In this short communication, we review the relationship between frailty and malnutrition risk in the elderly. Frailty is a term used for elderly at increased risk of adverse outcomes, including disability, falls, hospitalization, need for long-term care, and mortality. The Mini Nutritional Assessment (MNA) was designed and validated in a series of studies to assess nutritional status of elderly, as integral part of the comprehensive geriatric assessment, with a 2-steps screening process; when the MNA-SF classify a person at risk, the full MNA should be completed. The MNA and MNA-SF are sensitive, specific, and accurate in identifying nutrition risk. Increased risk of malnutrition, a common condition in the elderly, is closely associated with many potential contributors of frailty. The maintenance of optimal physical and cognitive performances depends on the early screening of critical conditions to develop preventive targeted interventions; the MNA supports such preventive action.


Epidemiological features of women with trichomoniasis in Auckland sexual health clinics: 1998-99.
To determine some epidemiological features of female sexually transmitted disease (STD) clinic attendees with Trichomonas vaginalis infection. A retrospective audit of patient charts was performed on all cases of T. vaginalis infection diagnosed in female patients in an 18-month period from January 1998 to June 1999 (n=88). Descriptive features of these cases were collated. The ethnicity of female patients with T. vaginalis was compared to that of all other female attendees within the same period. The incidence of T. vaginalis infection was 2.2% in 1998 1999. The mean age of patients was 26.5 years. Maori and Polynesian women were over-represented. Overall, patients were more likely to be symptomatic at presentation (78%) and to have abnormal findings on clinical examination (81%). 28% had co-existing chlamydia infection and 10% had co-existing gonorrhoea infection. The mean age of 26.5 years is lower than that quoted in other studies. The majority of patients had more than one condition at diagnosis. The rate of co-infection with chlamydia is high and some consideration could be given to empirical treatment of chlamydia in patients diagnosed with T. vaginalis infection.


Label-free based comparative proteomic analysis of whey proteins between different milk yields of Dezhou donkey.
Donkey milk, similar to human milk in compositions, has been suggested as the best potential hypoallergenic replacement diet for babies suffering from cow milk protein allergens and a promising nutraceutical for aged people. In this study, label-free mass spectrometry analysis was conducted to quantitatively identify the whey proteins differentially expressed in high-milk-yield samples compared with low-milk-yield samples. A total of 216 whey proteins were identified, and 19 of them showed significant differences in high-milk-yield samples. Of these proteins, 16 were upregulated and 3 were downregulated. Differentially expressed proteins (DEPs) were subjected to intensive bioinformatic analysis. Results revealed that the majority of DEPs participated in protein processing in endoplasmic reticulum, estrogen signaling pathway, progesterone-mediated oocyte maturation, and PI3K-Akt signaling pathway. Functional protein analysis suggested that proteins functioned in binding, catalytic activity, molecular function regulation, structural molecule activity, and transporter activity. Our study was the first to analyze the whey protein profile of different samples of donkey milk and to identify candidate proteins that could be used to explore the molecular mechanism related to the yield traits of Dezhou donkey milk. This study provided the biomarkers for the selection of high-milk-yielding donkey and obtained valuable information for future studies.


Neuromedin U directly stimulates growth of cultured rat calvarial osteoblast-like cells acting via the NMU receptor 2 isoform.
The neuromedin U (NMU) system is composed of NMU, neuromedin S (NMS) and their receptors NMUR1 and NMUR2. This system is involved in the regulation of energy homeostasis, neuroendocrine functions, immune response, circadian rhythm and spermatogenesis. The present study aimed to investigate the possible role of the NMU system in regulating functions of cultured rat calvarial osteoblast-like (ROB) cells. By using QPCR, high expression of NMU mRNA was found in freshly isolated ROB cells while after 7, 14, and 21 days of culture, expression of the studied gene was very low. In contrast, NMUR2 mRNA expression in freshly isolated ROB cells was negligible and very high in cultured cells. The highest NMUR2 mRNA expression was observed at day 7, and was followed by lower levels at days 14 and 21 of culture. Neither NMS nor NMUR1 mRNA was found in studied cells. Exposure of cultured ROB cells to NMU8 at concentrations 10(-6) to 10(-10) M had no effect on expression levels of the genes. During the entire culture period, NMU8 did not affect osteocalcin production, but stimulated proliferative activity of ROB cells at days 14 and 21 of culture. Thus, we demonstrated that cultured rat calvarial osteoblast-like cells are provided with NMUR2, the receptor isoform typical for the central nervous system. Acting via this receptor NMU8 stimulates proliferation of cultured cells and has no effect on their differentiated function (osteocalcin secretion).


Approach to a Standardized Injury Prevention Coordinator Training Curriculum.
The effectiveness of trauma center-based injury prevention programs is constrained by a number of variables. These include the wide range of injury types, the variety of possible interventions, institutional support and funding, and the knowledge and experience of the persons responsible for these programs. As the field of injury prevention has increased in complexity, so must the role and professional development of these injury prevention professionals responsible for these programs. Trauma center-based injury prevention coordinators are a diverse group with variable education and professional background especially related to public health, advocacy, epidemiology, biostatistics, and research. Furthermore, inconsistencies exist with their job titles, responsibilities, accountability, and authority, as well as the associated professional resources available to them. The American Trauma Society, with facilitation by the member organizations of the Trauma Prevention Coalition, has addressed the need to standardize the educational foundation for injury prevention coordinators by providing the basis of core competencies that are necessary to successfully oversee an American College of Surgeon's Committee on Trauma-verified trauma center's injury prevention program. This inaugural Injury Prevention Coordinators Course was launched in conjunction with the Society of Trauma Nurses 2015 annual conference in Jacksonville, FL, with 7 additional courses having been held through March 2016, comprising 150 participants. The goal of this 2-day, formal trauma center-based, course is to address and standardize key educational segments, including impact of trauma, program development, program evaluation, public health models, injury and data analysis, epidemiology, advocacy, building partnerships and coalitions, and the use of media promotion to ensure consistency throughout the industry.


Stabilizing Cu2+ Ions by Solid Solutions to Promote CO2 Electroreduction to Methane.
Copper is the only metal catalyst that can perform the electrocatalytic CO2 reduction reaction (CRR) to produce hydrocarbons and oxygenates. Its surface oxidation state determines the reaction pathway to various products. However, under the cathodic potential of CRR conditions, the chemical composition of most Cu-based catalysts inevitably undergoes electroreduction from Cu2+ to Cu0 or Cu1+ species, which is generally coupled with phase reconstruction and the formation of new active sites. Since the initial Cu2+ active sites are hard to retain, there have been few studies about Cu2+ catalysts for CRR. Herein we propose a solid-solution strategy to stabilize Cu2+ ions by incorporating them into a CeO2 matrix, which works as a self-sacrificing ingredient to protect Cu2+ active species. In situ spectroscopic characterization and density functional theory calculations reveal that compared with the conventionally derived Cu catalysts with Cu0 or Cu1+ active sites, the Cu2+ species in the solid solution (Cu-Ce-Ox) can significantly strengthen adsorption of the *CO intermediate, facilitating its further hydrogenation to produce CH4 instead of dimerization to give C2 products. As a result, different from most of the other Cu-based catalysts, Cu-Ce-Ox delivered a high Faradaic efficiency of 67.8% for CH4 and a low value of 3.6% for C2H4.


Seroepidemiology of the human T-cell leukaemia/lymphoma viruses in Europe. The HTLV European Research Network.
An extensive collaboration of laboratories and investigators has been developed to define the seroprevalence of human T-cell leukaemia/ lymphoma virus type I and II (HTLV-I and -II) infection in Europe. An algorithm for serological screening for HTLV-I and -II infection has been established by consensus. Data from screening almost 4 million subjects, including many unpublished studies, which conform to this algorithm are presented. In extensive studies the seroprevalence of HTL.V-I/II in blood donors is low, ranging from < 1 in 100,000 to 30 in 100,000 donors and is due predominantly to HTLV-I. In antenatal clinics in France and the United Kingdom the seroprevalence of HTLV-I is > 0.2%, but surveillance in this setting has been limited and extensive study of the seroprevalence of HTLV-I/II infection in pregnant women in Europe is urgently required to determine the need for HTLV-I/II antenatal screening. HTLV-I is present in populations who have immigrated to Europe from endemic areas and is spreading into indigenous European populations, particularly through sexual transmission to females. HTLV-II infection is present predominantly amongst IVDU and is usually a coinfection with HIV-I. There are considerable regional differences in HTLV-II seroprevalence.


Green synthesis strategy for producing doped and undoped ZnO nanoparticles: their photocatalytic studies for industrial dye degradation.
A facile and green method for synthesizing Zinc Oxide nanoparticles (ZnO NPs) was successfully carried out using unutilized sweet lime; i.e., Citrus Limetta rind pulp (U-CLRP) extract. The structural, morphological and optical studies were elucidated to confirm the crystallinity, size, and shape of the synthesized NPs. Copper doping on ZnO NPs (CZnO NPs) was carried out to enhance the optical properties. The as-prepared and doped nanoparticle's potential for efficient degradation of a commercial dye, Methylene Blue (MB), was tested under Ultraviolet (UV) and visible light radiation. ZnO NPs showed promising results for dye degradation while an improved result was witnessed for CZnO NPs. ZnO NPs showed 74% of degradation of MB dye under UV irradiation and 57% degradation under visible light radiation. CZnO NPs presented 85 and 90% degradation in the UV and visible radiation, respectively. This green reduction method utilizing biological waste sources shows a promising path for photocatalysts to be developed economically as well as efficiently in the future. These CZnO/ ZnO NPs proved their potential for embedding them as efficient catalysts for wastewater treatment plants of textile industries.


Physiological response of Pinus halepensis needles under ozone and water stress conditions.
The aim of this study was to evaluate how physiological processes of potted Pinus halepensis plants, grown under controlled conditions, were affected by ozone (O3) and/or water stress, integrating the gas exchange and biochemical data with fluorescence OJIP polyphasic transient data. Plants submitted to only water stress (T1) and with ozone (T3) showed a strong decrease in stomatal conductance and gas exchange, coinciding with a reduction of maximum yield of photochemistry (varphipo) and very negative values of leaf water potential. Simultaneously, a great increase of both PSII antenna size, indicated by absorption per reaction centre, and electron transport per reaction centre were found. The reduction of photosynthesis in the O3-treated plants (T2) by a slowing down of the Calvin cycle was supported by the increase of related fluorescence parameters such as relative variable fluorescence, heat de-excitation constant, energy de-excitation by spillover, and the decrease of varphipo. We suggest an antagonistic effect between the two stresses to explain the delayed ozone-induced decrease of stomatal conductance values for T3 with respect to T1 plants, by an alteration of the physiological mechanisms of stomatal opening, which involve the increase of intra-cellular free-calcium induced by ABA under co-occurring water shortage. We emphasise the importance of considering the intensity of the individual stress factor in studies concerning the interaction of stresses.


Endophilin B1 regulates EGFR endocytic degradation in prostate cancer cell.
Prostate cancer (Pca) is one of the most common types of cancer for elder men. Aberrant expression of epidermal growth factor receptor (EGFR) and EGFR downstream signaling have been known to contribute to disease progression in prostate cancer. EGF-stimulated EGFR is internalized and the process of endocytic degradation of EGFR mediates its signaling which is frequently dysregulated in many kinds of cancer. In the present study, we demonstrated that endophilin B1 expression was inhibited and EGFR expression was significantly increased in prostate cancer cell lines. We demonstrated that suppression of endophilin B1 increased EGFR levels via delaying EGFR internalization triggered by EGF and its intracellular degradation. Endophilin B1 decreased also sustained EGFR downstream signaling such as Erk1/2 phosphorylation in response to EGF stimulation and promoted prostate cancer cell proliferation which is EGF independent. Our data indicated that endophilin B1 mediated the biological function of EGFR in cancer cell proliferation through regulating the EGFR endocytic trafficking and downstream signaling.


Salinimonas lutimaris sp. nov., a polysaccharide-degrading bacterium isolated from a tidal flat.
A Gram-negative, non-motile, non-endospore-forming bacterial strain, designated DPSR-4(T), was isolated from a tidal flat sediment on the southern coast of Korea. Strain DPSR-4(T) grew optimally at 25-30°C, at pH 7.0-7.5 and in the presence of 2% (w/v) NaCl. A Neighbour-Joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain DPSR-4(T) clustered with Salinimonas chungwhensis BH030046(T) by a high bootstrap resampling value of 99.7%. Strain DPSR-4(T) exhibited 96.2% 16S rRNA gene sequence similarity to that of S. chungwhensis BH030046(T) and 93.7-96.6% sequence similarity to the sequences of type strains of Alteromonas species. Strain DPSR-4(T) contained Q-8 as the predominant ubiquinone and iso-C(15:0) 2-OH and/or C(16:1) ω7c, C(16:0) and C(18:1) ω7c as the major fatty acids. The major polar lipids detected in strain DPSR-4(T) and S. chungwhensis KCTC 12239(T) were phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. The DNA G+C content was 53.4 mol%. Differential phenotypic properties and phylogenetic distinctiveness of strain DPSR-4(T) demonstrated that this strain is distinguishable from the sole recognized species of the genus Salinimonas, S. chungwhensis. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain DPSR-4(T) is considered to represent a novel species of the genus Salinimonas, for which the name Salinimonas lutimaris sp. nov. is proposed. The type strain is DPSR-4(T) (KCTC 23464(T), CCUG 60743(T)).


Influence of storage conditions of extracted teeth on dentine removal by a standardised method of filing.
Extracted teeth are often stored in a variety of solutions for different lengths of time prior to use in studies on the efficacy of root canal preparation. The aim of this study was to evaluate the effect of duration (12 h, 1, 4 and 15 weeks) and type of storage medium (Saline, Formal-saline, Sodium Hypochlorite and Dry conditions) on dentine removal by standardised filing. Apparatus was constructed to standardise variables inherent in filing. These included the file, the rate, duration and amplitude of its push-pull motion, its alignment with a flat dentine surface, the interfacial force (80 gms) and rate of irrigation. Freshly extracted teeth were immediately stored in one of the 13 combinations of storage media and duration. The teeth were prepared into test specimens by splitting the roots and flattening the root canal surface. Twenty specimens were designated to each combination. Dentine removal was quantified by profilometry using maximum depth at 8 selected points. Statistical analysis of the mean values showed highly significant differences (p<0.001) between the media at 1, 4 and 15 weeks.When comparisons of mean values were made within the storage groups, there were highly significant differences (p<0.001) in dentine removal between 0, 1, 4 and 15 weeks of storage in sodium hypochlorite and saline. There were no significant differences found for storage in formal-saline. In dry conditions, the differences were significant (p<0.01) between 0, 1, 4 and 15 weeks of storage, but were not significant when the 4 weeks values were excluded.


Dinuclear oxidative addition of N-H and S-H bonds at chromium. Reaction of (*)Cr(CO)(3)(C(5)Me(5)) with [o-(HA)C(6)H(4)S-Cr(CO)(3)(C(5)Me(5))] (A = S, NH) yielding [eta(2)-o-(mu-A)C(6)H(4)S-Cr(C(5)Me(5))](2) and H-Cr(CO)(3)(C(5)Me(5)).
Reaction of the 17-electron radical (*)Cr(CO)(3)Cp* (Cp* = C(5)Me(5)) with 0.5 equiv of 2-aminophenyl disulfide [(o-H(2)NC(6)H(4))(2)S(2)] results in rapid oxidative addition to form the initial product (o-H(2)N)C(6)H(4)S-Cr(CO)(3)Cp*. Addition of a second equivalent of (*)Cr(CO)(3)Cp* to this solution results in the formation of H-Cr(CO)(3)Cp* as well as (1)/(2)[[eta(2)-o-(mu-NH)C(6)H(4)S]CrCp*](2). Spectroscopic data show that (o-H(2)N)C(6)H(4)S-Cr(CO)(3)Cp* loses CO to form [eta(2)-(o-H(2)N)C(6)H(4)S]Cr(CO)(2)Cp*. Attack on the N-H bond of the coordinated amine by (*)Cr(CO)(3)Cp* provides a reasonable mechanism consistent with the observation that both chelate formation and oxidative addition of the N-H bond are faster under argon than under CO atmosphere. The N-H bonds of uncoordinated aniline do not react with (*)Cr(CO)(3)Cp*. Reaction of the 2 mol of (*)Cr(CO)(3)Cp* with 1,2-benzene dithiol [1,2-C(6)H(4)(SH)(2)] yields the initial product (o-HS)C(6)H(4)S-Cr(CO)(3)Cp and 1 mol of H-Cr(CO)(3)Cp*. Addition of 1 equiv more of (*)Cr(CO)(3)Cp to this solution also results in the formation of 1 equiv of H-Cr(CO)(3)Cp*, as well as the dimeric product (1)/(2)[[eta(2)-o-(mu-S)C(6)H(4)S]CrCp*](2). This reaction also occurs more rapidly under Ar than under CO, consistent with intramolecular coordination of the second thiol group prior to oxidative addition. The crystal structures of [[eta(2)-o-(mu-NH)C(6)H(4)S]CrCp*](2) and [[eta(2)-o-(mu-S)C(6)H(4)S]CrCp*](2) are reported.


Decoding ubiquitin for mitosis.
Conjugation of ubiquitin (ubiquitination) to substrate proteins is a widespread modification that ensures fidelity of many cellular processes. During mitosis, different dynamic morphological transitions have to be coordinated in a temporal and spatial manner to allow for precise partitioning of the genetic material into two daughter cells, and ubiquitination of key mitotic factors is believed to provide both directionality and fidelity to this process. While directionality can be achieved by a proteolytic type of ubiquitination signal, the fidelity is often determined by various types of ubiquitin conjugation that does not target substrates for proteolysis by the proteasome. An additional level of complexity is provided by various ubiquitin-interacting proteins that act downstream of the ubiquitinated substrate and can serve as "decoders" for the ubiquitin signal. They may, specifically reverse ubiquitin attachment (deubiquitinating enzymes, DUBs) or, act as a receptor for transfer of the ubiquitinated substrate toward downstream signaling components and/or subcellular compartments (ubiquitin-binding proteins, UBPs). In this review, we aim at summarizing the knowledge and emerging concepts about the role of ubiquitin decoders, DUBs, and UBPs that contribute to faithful regulation of mitotic division.


Plant-Produced Chimeric VHH-sIgA Against Enterohemorrhagic E. coli Intimin Shows Cross-Serotype Inhibition of Bacterial Adhesion to Epithelial Cells.
Enterohemorrhagic Escherichia coli (EHEC) has consistently been one of the foremost foodborne pathogen threats worldwide based on the past 30 years of surveillance. EHEC primarily colonizes the bovine gastrointestinal (GI) tract from which it can be transmitted to nearby farm environments and remain viable for months. There is an urgent need for effective and easily implemented pre-harvest interventions to curtail EHEC contamination of the food and water supply. In an effort to address this problem, we isolated single-domain antibodies (VHHs) specific for intimin, an EHEC adhesin required for colonization, and designed chimeric VHH fusions with secretory IgA functionality intended for passive immunotherapy at the mucosal GI surface. The antibodies were produced in leaves of Nicotiana benthamiana with production levels ranging between 1 and 3% of total soluble protein. in vivo assembly of all subunits into a hetero-multimeric complex was verified by co-immunoprecipitation. Analysis of multivalent protection across the most prevalent EHEC strains identified one candidate antibody, VHH10-IgA, that binds O145:Hnm, O111:Hnm, O26:H11, and O157:H7. Fluorometric and microscopic analysis also indicated that VHH10-IgA completely neutralizes the capacity of the latter three strains to adhere to epithelial cells in vitro. This study provides proof of concept that a plant-produced chimeric secretory IgA can confer cross-serotype inhibition of bacterial adhesion to epithelial cells.


Percutaneous closure of post-traumatic and congenital muscular ventricular septal defects with the Amplatzer Muscular VSD Occluder.
Muscular ventricular septal defects (VSD) are an important and difficult surgical problem. In the last few years a new alternative has emerged - possibility of VSD closure using percutaneous approach. To present our experience in percutaneous closure of congenital muscular and one posttraumatic VSD. We treated 10 patients - 7 children (age 0.8-7 years) and 2 adults (43 and 46 years) with congenital VSD, and one 18-year-old patient with posttraumatic VSD (knife stab). All the patients had a large haemodynamic shunt (Qp:Qs 1.9) and in all cases percutaneous closure attempt with an Amplatzer Muscular VSD Occluder (MVSDO) implant was undertaken. Five of 6 children with multiple muscular VSDs had in infancy previous pulmonary artery banding and one patient had complex heart disease: transposition of great arteries (dTGA), pulmonary stenosis (PS) and perimembranous VSD. All procedures were performed using the standard technique. Eleven procedures were performed in 10 patients (one child had 2 attempts). Seven procedures were successful. In all cases a considerable reduction in flow or complete closure was achived. In one case, despite multiple attempts, VSD caniulation was ineffective and the procedure was abandoned. The patient had oblique VSD - morphology confirmed was later by the operating surgeon. The reason for the other 3 failures was early embolisation to the left ventricle and aorta. This complication was noted in 2 adult patients - one with congenital and one with post-traumatic VSD. In both cases the interventricular septum was thick (10 and 11 mm) and implants were removed with a bioptome or vascular lasso. Another embolisation occurred in a child with TGA - in this case the cardiac surgeon removed the implant from the aortic arch during Rastelli operation. Our experience acquired during muscular VSD closure with MVSDO indicates that the method is useful in children with isolated defects. Adult patients and children with a complex form of congenital defects should have morphology of MVSDO carefully evaluated and width of the interventricular septum measured to avoid potential implant embolisation.


Magnetic resonance microscopy of the barramundi (Lates calcarifer) brain.
Magnetic resonance imaging (MRI) is an established technique for morphological imaging of the central nervous system. Despite its prevalent use in a range of taxa, few studies exist on the brains of teleosts. In this study, we present a detailed analysis of a teleost brain using high-resolution MRI. Images were acquired from a Bruker 16.4 T vertical magnet with a three-dimensional flash T(2)*-weighted image sequence. High contrast was obtained using Magnevist® and the following imaging parameters: a flip angle of 30°, a repetition time of 50 ms, an echo time of 14 ms, and an image matrix of 1024 × 400 × 400. The resulting isotropic resolution of 30 μm allowed us to thoroughly describe the architecture of the barramundi (Lates calcarifer) brain, including descriptions of nuclei, fiber tracts, and cellular layers. A good correspondence, both in contrast and morphology, was found between magnetic resonance images and Nissl-stained brain sections, allowing for an analysis of the benefits and drawbacks of MRI and conventional histology.


Histidine-tagging and purification of tobacco etch potyvirus helper component protein.
The coding sequence for a series of six histidines (his-tag) was inserted near the 5' terminus of the helper component (HC) coding region of tobacco etch potyvirus (TEV). Full length genomic clones containing the his-tag coding sequence were infectious and produced symptoms in tobacco (Nicotiana tabacuma) similar to those induced by wild-type TEV. The modified virus was genetically stable and the his-tag sequence was maintained through at least four cycles of aphid transmission. A protocol for purifying rapidly the HC protein, based on the affinity of its his-tag for Ni(2+)-charged resin, yielded large amounts of his-tagged HC protein that was fully functional as demonstrated by aphid transmission experiments.