In vitro activity of cefotetan, a new cephamycin derivative, compared with that of other beta-lactam compounds.
The in vitro activity of cefotetan, a new cephamycin, was compared with the activities of cefoxitin, cefuroxime, moxalactam, ceftazidime, and piperacillin against 273 recent clinical isolates. The minimum inhibitory concentrations of cefotetan for 90% of Enterobacteriaceae, Haemophilus influenzae, and Neisseria gonorrhoeae were between 0.12 and 2 micrograms/ml; for 90% of Staphylococcus aureus, the minimum inhibitory concentration was 8 micrograms/ml, and for 90% of Bacteroides fragilis, Pseudomonas aeruginosa, and Lancefield group D streptococci it was 128 micrograms/ml or more. The activity of cefotetan against Enterobacteriaceae is comparable to the activities of ceftazidime and moxalactam and four to eight times greater than the activities of cefoxitin and cefuroxime. Cefotetan was approximately one-half as active as cefoxitin and cefuroxime against S. aureus. Although cefotetan was 87% bound to serum protein, serum had little effect on the in vitro antimicrobial activity of this agent.


α2δ-1 gene deletion affects somatosensory neuron function and delays mechanical hypersensitivity in response to peripheral nerve damage.
The α2δ-1 subunit of voltage-gated calcium channels is upregulated after sensory nerve injury and is also the therapeutic target of gabapentinoid drugs. It is therefore likely to play a key role in the development of neuropathic pain. In this study, we have examined mice in which α2δ-1 gene expression is disrupted, to determine whether α2δ-1 is involved in various modalities of nociception, and for the development of behavioral hypersensitivity after partial sciatic nerve ligation (PSNL). We find that naive α2δ-1(-/-) mice show a marked behavioral deficit in mechanical and cold sensitivity, but no change in thermal nociception threshold. The lower mechanical sensitivity is mirrored by a reduced in vivo electrophysiological response of dorsal horn wide dynamic range neurons. The CaV2.2 level is reduced in brain and spinal cord synaptosomes from α2δ-1(-/-) mice, and α2δ-1(-/-) DRG neurons exhibit lower calcium channel current density. Furthermore, a significantly smaller number of DRG neurons respond to the TRPM8 agonist menthol. After PSNL, α2δ-1(-/-) mice show delayed mechanical hypersensitivity, which only develops at 11 d after surgery, whereas in wild-type littermates it is maximal at the earliest time point measured (3 d). There is no compensatory upregulation of α2δ-2 or α2δ-3 after PSNL in α2δ-1(-/-) mice, and other transcripts, including neuropeptide Y and activating transcription factor-3, are upregulated normally. Furthermore, the ability of pregabalin to alleviate mechanical hypersensitivity is lost in PSNL α2δ-1(-/-) mice. Thus, α2δ-1 is essential for rapid development of mechanical hypersensitivity in a nerve injury model of neuropathic pain.


Performance of delayed-enhancement magnetic resonance imaging with gadoversetamide contrast for the detection and assessment of myocardial infarction: an international, multicenter, double-blinded, randomized trial.
The identification and assessment of myocardial infarction (MI) are important for therapeutic and prognostic purposes, yet current recommended diagnostic strategies have significant limitations. We prospectively tested the performance of delayed-enhancement magnetic resonance imaging (MRI) with gadolinium-based contrast for the detection of MI in an international, multicenter trial. Patients with their first MI were enrolled in an acute (< or = 16 days after MI; n=282) or chronic (17 days to 6 months; n=284) arm and then randomized to 1 of 4 doses of gadoversetamide: 0.05, 0.1, 0.2, or 0.3 mmol/kg. Standard delayed-enhancement MRI was performed before contrast (control) and 10 and 30 minutes after gadoversetamide. For blinded analysis, precontrast and postcontrast MRIs were randomized and then scored for enhanced regions by 3 independent readers not associated with the study. The infarct-related artery perfusion territory was scored from x-ray angiograms separately. In total, 566 scans were performed in 26 centers using commercially available scanners from all major US/European vendors. All scans were included in the analysis. The sensitivity of MRI for detecting MI increased with rising dose of gadoversetamide (P<0.0001), reaching 99% (acute) and 94% (chronic) after contrast compared with 11% before contrast. Likewise, the accuracy of MRI for identifying MI location (compared with infarct-related artery perfusion territory) increased with rising dose of gadoversetamide (P<0.0001), reaching 99% (acute) and 91% (chronic) after contrast compared with 9% before contrast. For gadoversetamide doses > or = 0.2 mmol/kg, 10- and 30-minute images provided equal performance, and peak creatine kinase-MB levels correlated with MRI infarct size (P<0.0001). Gadoversetamide-enhanced MRI using doses of > or = 0.2 mmol/kg is effective in the detection and assessment of both acute and chronic MI. This study represents the first multicenter trial designed to evaluate an imaging approach for detecting MI.


Detection of transient in vivo interactions between substrate and transporter during protein translocation into the endoplasmic reticulum.
The split-ubiquitin technique was used to detect transient protein interactions in living cells. Nub, the N-terminal half of ubiquitin (Ub), was fused to Sec62p, a component of the protein translocation machinery in the endoplasmic reticulum of Saccharomyces cerevisiae. Cub, the C-terminal half of Ub, was fused to the C terminus of a signal sequence. The reconstitution of a quasi-native Ub structure from the two halves of Ub, and the resulting cleavage by Ub-specific proteases at the C terminus of Cub, serve as a gauge of proximity between the two test proteins linked to Nub and Cub. Using this assay, we show that Sec62p is spatially close to the signal sequence of the prepro-alpha-factor in vivo. This proximity is confined to the nascent polypeptide chain immediately following the signal sequence. In addition, the extent of proximity depends on the nature of the signal sequence. Cub fusions that bore the signal sequence of invertase resulted in a much lower Ub reconstitution with Nub-Sec62p than otherwise identical test proteins bearing the signal sequence of prepro-alpha-factor. An inactive derivative of Sec62p failed to interact with signal sequences in this assay. These in vivo findings are consistent with Sec62p being part of a signal sequence-binding complex.


[The precise characterization and the crucial mechanism of NO-induced cytotoxicity].
NO is believed to be involved in neurotoxicity after various neuronal stresses. NO donors are toxic and cause changes in cellular morphology such as condensed and fragmented chromatin, shriveled nuclei, apoptotic bodies and membrane blebbing. These observations are consistent with the overall description of apoptosis. The crucial mechanism of NO-induced cytotoxicity is still unclear. Several mechanisms for NO-induced cytotoxicity in neurons have been proposed. It has been reported that NO enhances ADP-ribosylation or S-nitrosylation of an increasing number of proteins, and two of these proteins were identified as NO-target proteins. One is glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a key enzyme of glycolytic conversion, which is S-nitrosylated by NO inhibiting the enzyme activity. Hence, inhibition of GAPDH activity by NO would decrease the amount of ATP. NO also activates poly (ADP-ribose) polymerase (PARP) in the presence of DNA damage. The activation of PARP results in depletion of NAD and ATP. The energy depletion by NO could cause cell death. Recently, several factors such as Fas, the caspases (interleukin-1 beta-converting enzyme (ICE)-like proteases), Bcl-2 and the tumor suppressor gene product p53 have been shown to be involved in apoptotic cell death. We here discuss the crucial mechanisms of NO-induced cytotoxicity and also discuss recent findings about the protective effect of NO on cell death.


Aromaticity as the driving force for the stability of non-IPR endohedral metallofullerene Bingel-Hirsch adducts.
We have studied the relative stabilities of Bingel-Hirsch non-IPR endohedral metallofullerene monoadducts having one, two, or three adjacent pentagon pairs. The most stable addition always leads to an open adduct and never occurs on [5,5] bonds. Our results show that the thermodynamics of the addition is governed by the additive local aromaticity of the rings of the final adducts.


Genotyping of single nucleotide polymorphisms using the SNP-RFLP method.
Genetic polymorphisms, including single nucleotide polymorphisms (SNPs), are responsible for inter-individual variability in susceptibility to cancer and other disorders. Both environmental factors (e.g., smoking or carcinogen exposure) and genetic variation underlie the development of cancer; however, studies of twins suggest that genetic variation is more important. Hence, the identification of SNPs makes an important contribution to cancer research. In this study, 13 SNPs in 12 genes were genotyped in HEK 293 and HeLa cells using the simple and inexpensive SNP-RFLP method. Sanger sequencing was performed for one SNP to validate the SNP-RFLP results. Of the 13 SNPs, 10 were homozygous and three were heterozygous (rs10937405, rs12296850, and rs3814113) in HEK 293 cells, while 12 were homozygous and one was heterozygous (rs995030) in HeLa cells. The cells carried eight disease-associated risk alleles (32% of typed alleles), including rs2853677, rs995030, rs2736100, and rs6010620 in HEK 293 cells, and rs10937405, rs3814113, rs4767364, and rs6010620 in HeLa cells. Four SNP loci were homozygous for different alleles in each cell line, with HEK 293 cells having a CC genotype at rs2853677, GG at rs2736100 and rs4767364, and TT at rs3819197, while HeLa cells had TT genotypes at rs2853677 and rs2736100, AA at rs4767364, and CC at rs3819197. In conclusion, these results are potentially applicable for testing of novel gene therapeutic approaches in future experiments where the non-risk alleles of the eight identified risk alleles are substituted into HEK 293 or HeLa cells.


IndOR: a new statistical procedure to test for SNP-SNP epistasis in genome-wide association studies.
Epistasis is often cited as the biological mechanism carrying the missing heritability in genome-wide association studies. However, there is a very few number of studies reported in the literature. The low power of existing statistical methods is a potential explanation. Statistical procedures are also mainly based on the statistical definition of epistasis that prevents from detecting SNP-SNP interactions that rely on some classes of epistatic models. In this paper, we propose a new statistic, called IndOR for independence-based odds ratio, based on the biological definition of epistasis. We assume that epistasis modifies the dependency between the two causal SNPs, and we develop a Wald procedure to test such hypothesis. Our new statistic is compared with three statistical procedures in a large power study on simulated data sets. We use extensive simulations, based on 45 scenarios, to investigate the effect of three factors: the underlying disease model, the linkage disequilibrium, and the control-to-case ratio. We demonstrate that our new test has the ability to detect a wider range of epistatic models. Furthermore, our new statistical procedure is remarkably powerful when the two loci are linked and when the control-to-case ratio is higher than 1. The application of our new statistic on the Wellcome Trust Case Control Consortium data set on Crohn's disease enhances our results on simulated data. Our new test, IndOR, catches previously reported interaction with more power. Furthermore, a new combination of variant has been detected by our new test as significantly associated with Crohn's disease.


Brief report: genetics of alcoholic cirrhosis-GenomALC multinational study.
The risk of alcohol-related liver cirrhosis increases with increasing alcohol consumption, but many people with very high intake escape from liver disease. We postulate that susceptibility to alcoholic cirrhosis has a complex genetic component and propose that this can be dissected through a large and sufficiently powered genomewide association study (GWAS). The GenomALC Consortium comprises researchers from Australia, France, Germany, Switzerland, United Kingdom, and United States, with a joint aim of exploring the genetic and genomic basis of alcoholic cirrhosis. For this National Institutes of Health/National Institute on Alcohol Abuse and Alcoholism funded study, we are recruiting high-risk drinkers who are either cases (with alcoholic cirrhosis) or controls (drinking comparable amounts over similar time, but free of significant liver disease). Extensive phenotypic data are obtained using semistructured interviews and patient records, and blood samples are collected. We have successfully recruited 859 participants including 538 matched case-control samples as of September 2014, using study-specific inclusion-exclusion criteria and data collection protocols. Of these, 580 are cases (442 men and 138 women) and 279 are controls (205 men and 74 women). Duration of excessive drinking was slightly greater in cases than controls and was significantly less in women than men. Cases had significantly lower lifetime alcohol intake than controls. Both cases and controls had a high prevalence of reported parental alcohol problems, but cases were significantly more likely to report that a father with alcohol problems had died from liver disease (odds ratio 2.53, 95% confidence interval 1.31 to 4.87, p = 0.0055). Recruitment of participants for a GWAS of alcoholic cirrhosis has proved feasible across countries with multiple sites. Affected patients often consume less alcohol than unaffected ones, emphasizing the existence of individual vulnerability factors. Cases are more likely to report liver disease in a father with alcohol problems than controls, consistent with a potential genetic component to the risk of alcoholic cirrhosis.


Estimation of the force of infection and infectious period of skin sores in remote Australian communities using interval-censored data.
Prevalence of impetigo (skin sores) remains high in remote Australian Aboriginal communities, Fiji, and other areas of socio-economic disadvantage. Skin sore infections, driven primarily in these settings by Group A Streptococcus (GAS) contribute substantially to the disease burden in these areas. Despite this, estimates for the force of infection, infectious period and basic reproductive ratio-all necessary for the construction of dynamic transmission models-have not been obtained. By utilising three datasets each containing longitudinal infection information on individuals, we estimate each of these epidemiologically important parameters. With an eye to future study design, we also quantify the optimal sampling intervals for obtaining information about these parameters. We verify the estimation method through a simulation estimation study, and test each dataset to ensure suitability to the estimation method. We find that the force of infection differs by population prevalence, and the infectious period is estimated to be between 12 and 20 days. We also find that optimal sampling interval depends on setting, with an optimal sampling interval between 9 and 11 days in a high prevalence setting, and 21 and 27 days for a lower prevalence setting. These estimates unlock future model-based investigations on the transmission dynamics of skin sores.


Exploring New Graduate Nurses' Perceptions of Factors Influencing Job Satisfaction and Resignation Ideation During COVID-19.
Using an observational design, new graduate nurses' perceptions of job satisfaction and resignation ideation during COVID-19 were explored. Higher job satisfaction was associated with higher self-confidence, lower stress, and working in first choice of unit. A higher likelihood of resignation ideation was associated with coworker incivility, higher stress, and not working in first choice unit. New graduate nurses need support from nursing professional development practitioners for challenges faced throughout the transition-to-practice year.


Patterns of Self-care in Adults With Heart Failure and Their Associations With Sociodemographic and Clinical Characteristics, Quality of Life, and Hospitalizations: A Cluster Analysis.
Self-care is important in heart failure (HF) treatment, but patients may have difficulties and be inconsistent in its performance. Inconsistencies in self-care behaviors may mirror patterns of self-care in HF patients that are worth identifying to provide interventions tailored to patients. The aims of this study are to identify clusters of HF patients in relation to self-care behaviors and to examine and compare the profile of each HF patient cluster considering the patient's sociodemographics, clinical variables, quality of life, and hospitalizations. This was a secondary analysis of data from a cross-sectional study in which we enrolled 1192 HF patients across Italy. A cluster analysis was used to identify clusters of patients based on the European Heart Failure Self-care Behaviour Scale factor scores. Analysis of variance and χ test were used to examine the characteristics of each cluster. Patients were 72.4 years old on average, and 58% were men. Four clusters of patients were identified: (1) high consistent adherence with high consulting behaviors, characterized by younger patients, with higher formal education and higher income, less clinically compromised, with the best physical and mental quality of life (QOL) and lowest hospitalization rates; (2) low consistent adherence with low consulting behaviors, characterized mainly by male patients, with lower formal education and lowest income, more clinically compromised, and worse mental QOL; (3) inconsistent adherence with low consulting behaviors, characterized by patients who were less likely to have a caregiver, with the longest illness duration, the highest number of prescribed medications, and the best mental QOL; (4) and inconsistent adherence with high consulting behaviors, characterized by patients who were mostly female, with lower formal education, worst cognitive impairment, worst physical and mental QOL, and higher hospitalization rates. The 4 clusters identified in this study and their associated characteristics could be used to tailor interventions aimed at improving self-care behaviors in HF patients.


Alcohol and other drug use at school leavers' celebrations.
A significant proportion of adolescents who attend celebratory events often engage in substantial alcohol and other drug use. We examined patterns, influences and impacts of drug consumption at an end of schooling life celebration. Seventeen- to 18-year-old Australians who intended to attend (n = 541) and who attended the celebration (n = 405), respectively, completed pre- and post-event surveys. Males consumed 18.44 and females 13.24 Australian standard drinks on an average day during the school leavers' event. Compared with their last social event, there was greater alcohol (P < 0.0005) and ecstasy use (P < 0.046 for Day 1 and P < 0.008 on Day 3). However, the number of drinks consumed per hour appeared to be similar across contexts. Most (87%) experienced at least one negative outcome attributed to alcohol and other drug use. Safety strategies were frequently used and appeared to be protective against some of the most common harms (hangover, vomiting, black out and unprotected sex). The use of alcohol and other drugs at this celebratory event appears to be reflective of the greater than usual number of drinking hours that are available to participants. The use of safety strategies can be successful in mitigating some of the most common drug-related harms.


Cervical spinal cord multiple sclerosis: evaluation with 2D multi-echo recombined gradient echo MR imaging.
The two-dimensional multi-echo recombined gradient echo (MERGE) technique automatically acquires and sums multiple gradient echoes at various echo times in cervical spine magnetic resonance (MR) imaging. This technique increases the grey-white matter contrast within the spinal cord and should also improve the depiction of cervical cord lesions. The aim of this study was to qualitatively and quantitatively evaluate MERGE imaging compared with T2-weighted fast spin-echo (T2WFSE) imaging for depicting multiple sclerosis (MS) lesions in the cervical cord. Nineteen consecutive patients (10 males and 9 females; age range 22-62 years, mean age 43.6 years) with clinically diagnosed MS were examined with cervical spinal cord MR imaging at 3 T including both MERGE and T2WFSE imaging. Qualitative evaluation for MS lesion conspicuity was performed. The quantitative criterion utilized to compare MERGE imaging with T2WFSE imaging was the lesion-to-background contrast-to-noise ratio (CNR). MERGE imaging showed 79 lesions and missed 1 that was depicted on T2WFSE imaging. T2WFSE imaging showed 46 lesions and missed 34 that were depicted on MERGE imaging. MERGE imaging was markedly superior to T2WFSE imaging in rendering greater lesion conspicuity. In the quantitative evaluation, the lesion-to-background CNR upon MERGE imaging was significantly higher than that upon T2WFSE imaging (P < 0.001, paired t-test). MERGE imaging in the cervical spinal cord increases detection and conspicuity of MS lesions. Strong consideration should be given to utilizing axial MERGE images in the diagnosis and follow-up study of cervical cord MS.


The role of coenzyme Q10 in statin-associated myopathy: a systematic review.
Statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) are currently the most effective medications for reducing low-density lipoprotein cholesterol concentrations. Although generally safe, they have been associated with a variety of myopathic complaints. Statins block production of farnesyl pyrophosphate, an intermediate in the synthesis of ubiquinone or coenzyme Q10 (CoQ10). This fact, plus the role of CoQ10 in mitochondrial energy production, has prompted the hypothesis that statin-induced CoQ10 deficiency is involved in the pathogenesis of statin myopathy. We identified English language articles relating statin treatment and CoQ10 levels via a PubMed search through August 2006. Abstracts were reviewed and articles addressing the relationship between statin treatment and CoQ10 levels were examined in detail. Statin treatment reduces circulating levels of CoQ10. The effect of statin therapy on intramuscular levels of CoQ10 is not clear, and data on intramuscular CoQ10 levels in symptomatic patients with statin-associated myopathy are scarce. Mitochondrial function may be impaired by statin therapy, and this effect may be exacerbated by exercise. Supplementation can raise the circulating levels of CoQ10, but data on the effect of CoQ10 supplementation on myopathic symptoms are scarce and contradictory. We conclude that there is insufficient evidence to prove the etiologic role of CoQ10 deficiency in statin-associated myopathy and that large, well-designed clinical trials are required to address this issue. The routine use of CoQ10 cannot be recommended in statin-treated patients. Nevertheless, there are no known risks to this supplement and there is some anecdotal and preliminary trial evidence of its effectiveness. Consequently, CoQ10 can be tested in patients requiring statin treatment, who develop statin myalgia, and who cannot be satisfactorily treated with other agents. Some patients may respond, if only via a placebo effect.


Sodium niobate based hierarchical 3D perovskite nanoparticle clusters.
We report a microwave assisted synthesis of NaNbO3 perovskite mesocrystals with a hierarchical morphology formed by the self-assembly of nanoparticles in particle clusters. The synthesis method combines non-aqueous sol-gel synthesis and microwave heating in a single step process that allows us to isolate crystalline single phase NaNbO3 in few minutes. A detailed investigation of the effect of the reaction temperature on the crystallinity and morphology of the product was conducted. The synthesis stabilizes the unusual orthorhombic phase Pmma, a property that can be ascribed to the crystal size (24 nm). TEM and SAED analyses show that the hierarchical polycrystalline particles behave as single crystals, a feature related to a non-classical crystallization mechanism. Moreover, the optical bandgap of this NaNbO3 phase was estimated for the first time. The results suggest the potential of this synthetic procedure for the fast production of high quality tertiary oxide nanocrystals.


Controlled trial of pretest education approaches to enhance informed decision-making for BRCA1 gene testing.
In response to the isolation of the BRCA1 gene, a breast-ovarian cancer-susceptibility gene, biotechnology companies are already marketing genetic tests to health care providers and to the public. Initial studies indicate interest in BRCA1 testing in the general public and in populations at high risk. However, the optimal strategies for educating and counseling individuals have yet to be determined. Our goal was to evaluate the impact of alternate strategies for pretest education and counseling on decision-making regarding BRCA1 testing among women at low to moderate risk who have a family history of breast and/or ovarian cancer. A randomized trial design was used to evaluate the effects of education only (educational approach) and education plus counseling (counseling approach), as compared with a waiting-list (control) condition (n = 400 for all groups combined). The educational approach reviewed information about personal risk factors, inheritance of cancer susceptibility, the benefits, limitations, and risks of BRCA1 testing, and cancer screening and prevention options. The counseling approach included this information, as well as a personalized discussion of experiences with cancer in the family and the potential psychological and social impact of testing. Data on knowledge of inherited cancer and BRCA1 test characteristics, perceived risk, perceived benefits, limitations and risks of BRCA1 testing, and testing intentions were collected by use of structured telephone interviews at baseline and at 1-month follow-up. Provision of a blood sample for future testing served as a proxy measure of intention to be tested (in the education and counseling arms of the study). The effects of intervention group on study outcomes were evaluated by use of hierarchical linear regression modeling and logistic regression modeling (for the blood sample outcome). All P values are for two-sided tests. The educational and counseling approaches both led to significant increases in knowledge, relative to the control condition (P < .001 for both). The counseling approach, but not the educational approach, was superior to the control condition in producing significant increases in perceived limitations and risks of BRCA1 testing (P < .01) and decreases in perceived benefits (P < .05). However, neither approach produced changes in intentions to have BRCA1 testing. Prior to and following both education only and education plus counseling, approximately one half of the participants stated that they intended to be tested; after the session, 52% provided a blood sample. Standard educational approaches may be equally effective as expanded counseling approaches in enhancing knowledge. Since knowledge is a key aspect of medical decision-making, standard education may be adequate in situations where genetic testing must be streamlined. On the other hand, it has been argued that optimal decision-making requires not only knowledge, but also a reasoned evaluation of the positive and negative consequences of alternate decisions. Although the counseling approach is more likely to achieve this goal, it may not diminish interest in testing, even among women at low to moderate risk. Future research should focus on the merits of these alternate approaches for subgroups of individuals with different backgrounds who are being counseled in the variety of settings where BRCA1 testing is likely to be offered.


Synthesis of Terminal Ribose Analogues of Adenosine 5'-Diphosphate Ribose as Probes for the Transient Receptor Potential Cation Channel TRPM2.
TRPM2 (transient receptor potential cation channel, subfamily M, member 2) is a nonselective cation channel involved in the response to oxidative stress and in inflammation. Its role in autoimmune and neurodegenerative diseases makes it an attractive pharmacological target. Binding of the nucleotide adenosine 5'-diphosphate ribose (ADPR) to the cytosolic NUDT9 homology (NUDT9 H) domain activates the channel. A detailed understanding of how ADPR interacts with the TRPM2 ligand binding domain is lacking, hampering the rational design of modulators, but the terminal ribose of ADPR is known to be essential for activation. To study its role in more detail, we designed synthetic routes to novel analogues of ADPR and 2'-deoxy-ADPR that were modified only by removal of a single hydroxyl group from the terminal ribose. The ADPR analogues were obtained by coupling nucleoside phosphorimidazolides to deoxysugar phosphates. The corresponding C2″-based analogues proved to be unstable. The C1″- and C3″-ADPR analogues were evaluated electrophysiologically by patch-clamp in TRPM2-expressing HEK293 cells. In addition, a compound with all hydroxyl groups of the terminal ribose blocked as its 1″-β- O-methyl-2″,3″- O-isopropylidene derivative was evaluated. Removal of either C1″ or C3″ hydroxyl groups from ADPR resulted in loss of agonist activity. Both these modifications and blocking all three hydroxyl groups resulted in TRPM2 antagonists. Our results demonstrate the critical role of these hydroxyl groups in channel activation.


High-dose ifosfamide is associated with severe, reversible cardiac dysfunction.
To determine the incidence and characterize the occurrence of cardiac toxicity with high-dose ifosfamide. Retrospective chart review. Biomedical research referral center. Fifty-two consecutive patients with advanced lymphoma or carcinoma enrolled in phase I trials of high-dose ifosfamide as part of combination chemotherapy with autologous bone marrow transplantation. Patients were given escalating doses (10 to 18 g/m2) of ifosfamide in combination with carboplatin and etoposide or with lomustine and vinblastine. The chart review focused on clinical, radiographic, or electrocardiographic evidence of cardiovascular dysfunction. Data from invasive hemodynamic monitoring, radionuclide cineangiography, and echocardiography were also reviewed. Nine of the patients treated with ifosfamide developed congestive heart failure (17%; 95% Cl, 8% to 30%). Eight of these patients, experiencing dyspnea, tachycardia, weight gain, and signs of pulmonary edema, required admission to an intensive care unit. Left ventricular contractility was found to be depressed when evaluated by radionuclide cineangiography, echocardiography, or both. Most patients responded to diuretic, vasodilator, and inotropic therapies. Two patients developed malignant ventricular arrhythmias. One patient died of intractable cardiogenic shock. Five patients died of multiorgan failure, despite showing improvement in left ventricular ejection fraction. Three patients survived and regained baseline left ventricular ejection fraction. High-dose ifosfamide is associated with severe but usually reversible myocardial depression and malignant arrhythmias.


Safety and convenience of once-weekly somapacitan in adult GH deficiency: a 26-week randomized, controlled trial.
Somapacitan is a reversible albumin-binding growth hormone (GH) derivative, developed for once-weekly administration. This study aimed to evaluate the safety of once-weekly somapacitan vs once-daily Norditropin®. Local tolerability and treatment satisfaction were also assessed. 26-week randomized, controlled phase 3 safety and tolerability trial in six countries (Nbib2382939). Male or female patients aged 18-79 years with adult GH deficiency (AGHD), treated with once-daily GH for ≥6 months, were randomized to once-weekly somapacitan (n = 61) or once-daily Norditropin (n = 31) administered subcutaneously by pen. Both treatments were dose titrated for 8 weeks to achieve insulin-like growth factor I (IGF-I) standard deviation score (SDS) levels within the normal range, and then administered at a fixed dose. Outcome measures were adverse events (AEs), including injection site reactions; occurrence of anti-somapacitan/anti-GH antibodies and change in treatment satisfaction, assessed using the Treatment Satisfaction Questionnaire for Medication-9 (TSQM-9). Mean IGF-I SDS remained between 0 and 2 SDS throughout the trial in both groups. AEs were mostly mild or moderate and transient in nature. The most common AEs were nasopharyngitis, headache and fatigue in both groups. More than 1500 somapacitan injections were administered and no clinically significant injection site reactions were reported. No anti-somapacitan or anti-GH antibodies were detected. The TSQM-9 score for convenience increased significantly more with somapacitan vs Norditropin (P = 0.0171). In this 26-week trial in patients with AGHD, somapacitan was well tolerated and no safety issues were identified. Once-weekly somapacitan was reported to be more convenient than once-daily Norditropin.


Segmentation and visualization of a large, high-resolution micro-CT data of mice.
High-resolution large datasets were acquired to improve the understanding of murine bone physiology. The purpose of this work is to present the challenges and solutions in segmenting and visualizing bone in such large datasets acquired using micro-CT scan of mice. The analyzed dataset is more than 50 GB in size with more than 6,000 2,048 × 2,048 slices. The study was performed to automatically measure the bone mineral density (BMD) of the entire skeleton. A global Renyi entropy (GREP) method was initially used for bone segmentation. This method consistently oversegmented skeletal region. A new method called adaptive local Renyi entropy (ALREP) is proposed to improve the segmentation results. To study the efficacy of the ALREP, manual segmentation was performed. Finally, a specialized high-end remote visualization system along with the software, VirtualGL, was used to perform remote rendering of this large dataset. It was determined that GREP overestimated the bone cross-section by around 30 % compared with ALREP. The manual segmentation process took 6,300 min for 6,300 slices while ALREP took only 150 min for segmentation. Automatic image processing with ALREP method may facilitate BMD measurement of the entire skeleton in a significantly reduced time, compared with manual process.


The Streptococcus mutans IrvR repressor is a CI-like regulator that functions through autocleavage and Clp-dependent proteolysis.
Previous work has shown that irvR is required for the proper regulation of genetic competence and dextran-dependent aggregation due to its ability to repress the transcription regulator irvA. In this study, we determined the mechanism used to relieve the repression of irvA. We demonstrate that IrvR is a "LexA-like" protein with four conserved amino acid residues likely required for IrvR autocleavage activity. Furthermore, recombinant IrvR protein purified from Escherichia coli was competent to undergo autocleavage in vitro. Using several truncated IrvR constructs, we show that the amino acids adjacent to the autocleavage site are essential for relieving irvA repression and engaging the irvA-dependent regulatory pathway primarily through the ClpXP and ClpCP proteases. By extending the IrvR C terminus with an epitope derived from the autocleavage site, we were also able to create a constitutive Clp-dependent degradation of the full-length IrvR protein. This suggests that the derepression of irvA occurs through a two-step mechanism involving the initial autocleavage of IrvR and exposure of a proteolytic degradation sequence followed by Clp-dependent degradation of the IrvR DNA binding domain. Thus, irvA derepression is highly analogous to the genetic switch mechanism used to regulate lysogeny in bacteriophages.


ICON: Learning Regular Maps Through Inverse Consistency.
Learning maps between data samples is fundamental. Applications range from representation learning, image translation and generative modeling, to the estimation of spatial deformations. Such maps relate feature vectors, or map between feature spaces. Well-behaved maps should be regular, which can be imposed explicitly or may emanate from the data itself. We explore what induces regularity for spatial transformations, e.g., when computing image registrations. Classical optimization-based models compute maps between pairs of samples and rely on an appropriate regularizer for well-posedness. Recent deep learning approaches have attempted to avoid using such regularizers altogether by relying on the sample population instead. We explore if it is possible to obtain spatial regularity using an inverse consistency loss only and elucidate what explains map regularity in such a context. We find that deep networks combined with an inverse consistency loss and randomized off-grid interpolation yield well behaved, approximately diffeomorphic, spatial transformations. Despite the simplicity of this approach, our experiments present compelling evidence, on both synthetic and real data, that regular maps can be obtained without carefully tuned explicit regularizers, while achieving competitive registration performance.


Challenges to the provision of diabetes care in first nations communities: results from a national survey of healthcare providers in Canada.
Aboriginal peoples globally, and First Nations peoples in Canada particularly, suffer from high rates of type 2 diabetes and related complications compared with the general population. Research into the unique barriers faced by healthcare providers working in on-reserve First Nations communities is essential for developing effective quality improvement strategies. In Phase I of this two-phased study, semi-structured interviews and focus groups were held with 24 healthcare providers in the Sioux Lookout Zone in north-western Ontario. A follow-up survey was conducted in Phase II as part of a larger project, the Canadian First Nations Diabetes Clinical Management and Epidemiologic (CIRCLE) study. The survey was completed with 244 healthcare providers in 19 First Nations communities in 7 Canadian provinces, representing three isolation levels (isolated, semi-isolated, non-isolated). Interviews, focus groups and survey questions all related to barriers to providing optimal diabetes care in First Nations communities. the key factors emerging from interviews and focus group discussions were at the patient, provider, and systemic level. Survey results indicated that, across three isolation levels, healthcare providers' perceived patient factors as having the largest impact on diabetes care. However, physicians and nurses were more likely to rank patient factors as having a large impact on care than community health representatives (CHRs) and physicians were significantly less likely to rank patient-provider communication as having a large impact than CHRs. Addressing patient factors was considered the highest impact strategy for improving diabetes care. While this may reflect "patient blaming," it also suggests that self-management strategies may be well-suited for this context. Program planning should focus on training programs for CHRs, who provide a unique link between patients and clinical services. Research incorporating patient perspectives is needed to complete this picture and inform quality improvement initiatives.


Laboratory cultivation of widespread and previously uncultured soil bacteria.
Most soil bacteria belong to family-level phylogenetic groups with few or no known cultivated representatives. We cultured a collection of 350 isolates from soil by using simple solid media in petri dishes. These isolates were assigned to 60 family-level groupings in nine bacterial phyla on the basis of a comparative analysis of their 16S rRNA genes. Ninety-three (27%) of the isolates belonged to 20 as-yet-unnamed family-level groupings, many from poorly studied bacterial classes and phyla. They included members of subdivisions 1, 2, 3, and 4 of the phylum Acidobacteria, subdivision 3 of the phylum Verrucomicrobia, subdivision 1 of the phylum Gemmatimonadetes, and subclasses Acidimicrobidae and Rubrobacteridae of the phylum ACTINOBACTERIA: In addition, members of 10 new family-level groupings of subclass Actinobacteridae of the phylum Actinobacteria and classes Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria of the phylum Proteobacteria were obtained. The high degree of phylogenetic novelty and the number of isolates affiliated with so-called unculturable groups show that simple cultivation methods can still be developed further to obtain laboratory cultures of many phylogenetically novel soil bacteria.


Evidence of genotypic adaptation to the exposure to volcanic risk at the dopamine receptor DRD4 locus.
Humans have colonized and adapted to extremely diverse environments, and the genetic basis of some such adaptations, for example to high altitude, is understood. In some cases, local or regional variation in selection pressure could also cause behavioural adaptations. Numerous genes influence behaviour, such as alleles at the dopamine receptor locus D4 (DRD4), which are associated with attitude toward risk in experimental settings. We demonstrate genetic differentiation for this gene, but not for five unlinked microsatellite loci, between high- and low risk environments around Mount Merapi, an active volcano in Java, Indonesia. Using a behavioural experiment, we further show that people inhabiting the high risk environment are significantly more risk averse. We provide evidence of a genetic basis for this difference, showing that heterozygotes at the DRD4 locus are more risk averse than either homozygotes. In the high risk environment, allele frequencies are equilibrated, generating a high frequency of heterozygotes. Thus it appears that overdominance (i.e. selective advantage of heterozygotes) generates negative frequency dependent selection, favouring the rarer allele at this locus. Our results therefore provide evidence for adaptation to a marginal habitat through the selection of a neurocognitive trait with a genetic basis.


Apoptosis-induced CXCL5 accelerates inflammation and growth of prostate tumor metastases in bone.
During tumor progression, immune system phagocytes continually clear apoptotic cancer cells in a process known as efferocytosis. However, the impact of efferocytosis in metastatic tumor growth is unknown. In this study, we observed that macrophage-driven efferocytosis of prostate cancer cells in vitro induced the expression of proinflammatory cytokines such as CXCL5 by activating Stat3 and NF-κB(p65) signaling. Administration of a dimerizer ligand (AP20187) triggered apoptosis in 2 in vivo syngeneic models of bone tumor growth in which apoptosis-inducible prostate cancer cells were either coimplanted with vertebral bodies, or inoculated in the tibiae of immunocompetent mice. Induction of 2 pulses of apoptosis correlated with increased infiltration of inflammatory cells and accelerated tumor growth in the bone. Apoptosis-induced tumors displayed elevated expression of the proinflammatory cytokine CXCL5. Likewise, CXCL5-deficient mice had reduced tumor progression. Peripheral blood monocytes isolated from patients with bone metastasis of prostate cancer were more efferocytic compared with normal controls, and CXCL5 serum levels were higher in metastatic prostate cancer patients relative to patients with localized prostate cancer or controls. Altogether, these findings suggest that the myeloid phagocytic clearance of apoptotic cancer cells accelerates CXCL5-mediated inflammation and tumor growth in bone, pointing to CXCL5 as a potential target for cancer therapeutics.


Complications of endoscopy of the upper gastrointestinal tract: a single-center experience.
To evaluate prospectively the complications that occurred during consecutive endoscopies of the upper gastrointestinal tract. We evaluated all endoscopies of the upper gastrointestinal tract (except endoscopic retrograde cholangiopancreatography and endosonography) performed at the Ambulatory Surgical Center at the Mayo Clinic in Jacksonville, Fla, between January 1999 and June 2002. A staff gastroenterologist with or without a trainee performed these procedures. Therapeutic procedures included esophageal band ligation, injection sclerotherapy, botulinum toxin injection, extended upper endoscopy, pneumatic balloon dilation, endoscopic mucosal resection, and endoscopic ablation using thermal laser, argon beam coagulator, or photodynamic therapy. All complications were tabulated prospectively as per mandatory state licensure reporting. Complications after diagnostic endoscopy of the upper gastrointestinal tract were related to anesthesia in 2 of the 12,841 patients. Perforations in 5 patients were associated with esophageal dilation (2), resection of duodenal lesions (2), or passage of a side-viewing instrument into the duodenum (1). No deaths occurred. Diagnostic endoscopy of the upper gastrointestinal tract is safe, with a complication rate of less than 1 per 5000 cases. Therapeutic endoscopy increases the risk of complications. Compared with complication rates published previously, our results from a single center indicate a favorable reduction in complications related to endoscopy of the upper gastrointestinal tract.


A comparative assessment of mandible shape in a consomic strain panel of the house mouse (Mus musculus)--implications for epistasis and evolvability of quantitative traits.
Expectations of repeatedly finding associations between given genes and phenotypes have been borne out by studies of parallel evolution, especially for traits involving absence or presence of characters. However, it has rarely been asked whether the genetic basis of quantitative trait variation is conserved at the intra- or even at the interspecific level. This question is especially relevant for shape, where the high dimensionality of variation seems to require a highly complex genetic architecture involving many genes. We analyse here the genetic effects of chromosome substitution strains carrying M. m. musculus chromosomes in a largely M. m. domesticus background on mandible shape and compare them to the results of previously published QTL mapping data between M. m. domesticus strains. We find that the distribution of genetic effects and effect sizes across the genome is consistent between the studies, while the specific shape changes associated with the chromosomes are different. We find also that the sum of the effects from the different M. m. musculus chromosomes is very different from the shape of the strain from which they were derived, as well as all known wild type shapes. Our results suggest that the relative chromosome-wide effect sizes are comparable between the long separated subspecies M. m. domesticus and M. m. musculus, hinting at a relative stability of genes involved in this complex trait. However, the absolute effect sizes and the effect directions may be allele-dependent, or are context dependent, i.e. epistatic interactions appear to play an important role in controlling shape.


Psychosocial issues in space: results from Shuttle/Mir.
Important psychosocial issues involving tension, cohesion, leader support, and displacement of negative emotions were evaluated in a 4 1/2-year study involving five U.S. and four Russian Shuttle/Mir space missions. Weekly mood and group climate questionnaires were completed by five U.S. astronauts, eight Russian cosmonauts, and 42 U.S. and 16 Russian mission control subjects. There were few findings that supported our hypothesized changes in tension, cohesion, and leader support in crew and ground subjects using various time models, although crewmembers reported decreasing leader support in the 2nd half of the missions, and astronauts showed some evidence of a novelty effect in the first few weeks. There was no evidence suggesting a 3rd quarter effect among crewmembers on any of the 21 subscales evaluated. In contrast, there was strong evidence to support the hypothesized displacement of tension and negative emotions from crewmembers to mission control personnel and from mission control personnel to management. There were several significant differences in response between Americans vs. Russians, crewmembers vs. mission control personnel, and subjects in this study vs. people in comparable groups on Earth. Subject responses before, during, and after the missions were similar, and we did not find evidence for asthenia in space. Critical incidents that were reported generally dealt with events on-board the Mir and interpersonal conflicts, although most of the responses were from a relatively small number of subjects. Our findings have implications for future training and lead to a number of countermeasures.


High seasonal variation in entomologic inoculation rates in Eritrea, a semi-arid region of unstable malaria in Africa.
Entomologic studies were conducted in eight villages to investigate the patterns of malaria transmission in different ecologic zones in Eritrea. Mosquito collections were conducted for 24 months between September 1999 and January 2002. The biting rates of Anopheles arabiensis were highly seasonal, with activity concentrated in the wet season between June and October in the highlands and western lowlands, and between December and March in the coastal region. The biting rates in the western lowlands were twice as high as in the western escarpment and 20 times higher than in the coastal region. Sporozoite rates were not significantly different among villages. The risk of infection ranged from zero on the coast to 70.6 infective bites per year in the western lowlands. The number of days it would take for an individual to receive an infective bite from an infected An. arabiensis was variable among villages (range = 2.8-203.1 days). The data revealed the presence of only one main malaria transmission period between July and October for the highlands and western lowlands. Peak inoculation rates were recorded in August and September (range = 0.29-43.6 infective bits/person/month) at all sites over the two-year period. The annual entomologic inoculation rates (EIRs) varied greatly depending on year. The EIR profiles indicated that the risk of exposure to infected mosquitoes is highly heterogeneous and seasonal, with high inoculation rates during the rainy season, and with little or no transmission during the dry season. This study demonstrates the need to generate spatial and temporal data on transmission intensity on smaller scales to guide targeted control of malaria operations in semi-arid regions. Furthermore, EIR estimates derived in the present study provide a means of quantifying levels of exposure to infected mosquitoes in different regions of the country and could be important for evaluating the efficacy of vector control measures, since Eritrea has made significant steps in reducing the burden of malaria based on the Roll Back Malaria initiative of the World Health Organization.


Research ethics committees: Need for harmonization at the national level, the global and Indian perspective.
Ethics committee (EC) organization and standardization is an important aspect of clinical research. There is a healthy trend worldwide to register and/or accredit research ECs reviewing clinical research. This article tries to focus on the existing model of ECs worldwide, as against the Indian backdrop. The article reviews literature, journals, websites, and studies conducted in 10 different countries and outlines the working model of ECs in these countries. The challenges faced during the ethical review, especially in case of multicenter trials, have been identified. A solution has been suggested to overcome these challenges, and to ensure the overall smooth functioning of clinical trials. The article proposes the development of national and regional central ECs to counter the current drawbacks in the ethical review mechanisms in India.


CenteringPregnancy: an integrative literature review.
A critical analysis of the CenteringPregnancy model of care research has not been conducted across studies. The aims of this project were to describe 1) the conceptual components of the CenteringPregnancy practice model, 2) the characteristics of the CenteringPregnancy literature, and 3) the research methods and outcomes across the CenteringPregnancy research literature. An integrative literature review methodology was used to assess the CenteringPregnancy literature published between 1994 and 2009. The sample consisted of 26 articles (14 narrative descriptions, 10 quantitative studies, 1 mixed methods study, and 1 qualitative study) published predominantly in nursing and midwifery journals. The majority of quantitative studies were exploratory or quasi-experimental and longitudinal designs with the exception of 1 large, federally funded, randomized, controlled trial. Variables were individual and community level outcomes, which were not uniformly operationally defined or measured consistently across studies. Significant results for prenatal care attendance rates, adequate prenatal care, and breastfeeding rates were reported in a number of studies. Other results were often mixed or nonsignificant. Positive outcomes for the CenteringPregnancy model have been suggested by research findings to date but require more systematic approaches to developing knowledge about the factors inherent in this model that promote participant behavior changes, which lead to better perinatal outcomes as well as circumstances that maximize the effectiveness of this model. Areas for improving the strength of the evidence are discussed. Building consistently demonstrated relationships across studies is essential for creating the evidence base on which practice decisions can be made. Further research is required for developing clarity and understanding about this model's effectiveness.


Identification of Piccolo as a regulator of behavioral plasticity and dopamine transporter internalization.
Dopamine transporter (DAT) internalization is a mechanism underlying the decreased dopamine reuptake caused by addictive drugs like methamphetamine (METH). We found that Piccolo, a presynaptic scaffolding protein, was overexpressed in the nucleus accumbens (NAc) of the mice repeatedly administrated with METH. Piccolo downexpression by antisense technique augmented METH-induced behavioral sensitization, conditioned reward and synaptic dopamine accumulation in NAc. Expression of Piccolo C2A domain attenuated METH-induced inhibition of dopamine uptake in PC12 cells expressing human DAT. Consistent with this, it slowed down the accelerated DAT internalization induced by METH, thus maintaining the presentation of plasmalemmal DAT. In immunostaining and structural modeling Piccolo C2A domain displays an unusual feature of sequestering membrane phosphatidylinositol 4,5-bisphosphate, which may underlie its role in modulating DAT internalization. Together, our results indicate that Piccolo upregulation induced by METH represents a homeostatic response in the NAc to excessive dopaminergic transmission. Piccolo C2A domain may act as a cytoskeletal regulator for plasmalemmal DAT internalization, which may underlie its contributions in behavioral plasticity.


Crystal structure of HIV-1 gp41 including both fusion peptide and membrane proximal external regions.
The HIV-1 envelope glycoprotein (Env) composed of the receptor binding domain gp120 and the fusion protein subunit gp41 catalyzes virus entry and is a major target for therapeutic intervention and for neutralizing antibodies. Env interactions with cellular receptors trigger refolding of gp41, which induces close apposition of viral and cellular membranes leading to membrane fusion. The energy released during refolding is used to overcome the kinetic barrier and drives the fusion reaction. Here, we report the crystal structure at 2 A resolution of the complete extracellular domain of gp41 lacking the fusion peptide and the cystein-linked loop. Both the fusion peptide proximal region (FPPR) and the membrane proximal external region (MPER) form helical extensions from the gp41 six-helical bundle core structure. The lack of regular coiled-coil interactions within FPPR and MPER splay this end of the structure apart while positioning the fusion peptide towards the outside of the six-helical bundle and exposing conserved hydrophobic MPER residues. Unexpectedly, the section of the MPER, which is juxtaposed to the transmembrane region (TMR), bends in a 90 degrees-angle sideward positioning three aromatic side chains per monomer for membrane insertion. We calculate that this structural motif might facilitate the generation of membrane curvature on the viral membrane. The presence of FPPR and MPER increases the melting temperature of gp41 significantly in comparison to the core structure of gp41. Thus, our data indicate that the ordered assembly of FPPR and MPER beyond the core contributes energy to the membrane fusion reaction. Furthermore, we provide the first structural evidence that part of MPER will be membrane inserted within trimeric gp41. We propose that this framework has important implications for membrane bending on the viral membrane, which is required for fusion and could provide a platform for epitope and lipid bilayer recognition for broadly neutralizing gp41 antibodies.


Recommendations for collection and handling of specimens from group breast cancer clinical trials.
Recommendations for specimen collection and handling have been developed for adoption across breast cancer clinical trials conducted by the Breast International Group (BIG)-sponsored Groups and the National Cancer Institute (NCI)-sponsored North American Cooperative Groups. These recommendations are meant to promote identifiable standards for specimen collection and handling within and across breast cancer trials, such that the variability in collection/handling practices that currently exists is minimized and specimen condition and quality are enhanced, thereby maximizing results from specimen-based diagnostic testing and research. Three working groups were formed from the Cooperative Group Banking Committee, BIG groups, and North American breast cancer cooperative groups to identify standards for collection and handling of (1) formalin-fixed, paraffin-embedded (FFPE) tissue; (2) blood and its components; and (3) fresh/frozen tissue from breast cancer trials. The working groups collected standard operating procedures from multiple group specimen banks, administered a survey on banking practices to those banks, and engaged in a series of discussions from 2005 to 2007. Their contributions were synthesized into this document, which focuses primarily on collection and handling of specimens to the point of shipment to the central bank, although also offers some guidance to central banks. Major recommendations include submission of an FFPE block, whole blood, and serial serum or plasma from breast cancer clinical trials, and use of one fixative and buffer type (10% neutral phosphate-buffered formalin, pH 7) for FFPE tissue across trials. Recommendations for proper handling and shipping were developed for blood, serum, plasma, FFPE, and fresh/frozen tissue.


Eicosapentaenoic acid and docosahexaenoic acid from fish oils: differential associations with lipid responses.
Fish-oil supplementation can reduce circulating triacylglycerol (TG) levels and cardiovascular risk. This study aimed to assess independent associations between changes in platelet eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and fasting and postprandial (PP) lipoprotein concentrations and LDL oxidation status, following fish-oil intervention. Fifty-five mildly hypertriacylglycerolaemic (TG 1.5-4.0 mmol/l) men completed a double-blind placebo controlled cross over study, where individuals consumed 6 g fish oil (3 g EPA+DHA) or 6 g olive oil (placebo)/d for two 6-week intervention periods, with a 12-week wash-out period in between. Fish-oil intervention resulted in a significant increase in the platelet phospholipid EPA (+491 %, P<0.001) and DHA (+44 %, P<0.001) content and a significant decrease in the arachidonic acid (-10 %, P<0.001) and gamma-linolenic acid (-24 %, P<0.001) levels. A 30 % increase in ex vivo LDL oxidation (P<0.001) was observed. In addition, fish oil resulted in a significant decrease in fasting and PP TG levels (P<0.001), PP non-esterified fatty acid (NEFA) levels, and in the percentage LDL as LDL-3 (P=0.040), and an increase in LDL-cholesterol (P=0.027). In multivariate analysis, changes in platelet phospholipid DHA emerged as being independently associated with the rise in LDL-cholesterol, accounting for 16 % of the variability in this outcome measure (P=0.030). In contrast, increases in platelet EPA were independently associated with the reductions in fasting (P=0.046) and PP TG (P=0.023), and PP NEFA (P=0.015), explaining 15-20 % and 25 % of the variability in response respectively. Increases in platelet EPA+DHA were independently and positively associated with the increase in LDL oxidation (P=0.011). EPA and DHA may have differential effects on plasma lipids in mildly hypertriacylglycerolaemic men.


Genome-wide analysis of Polycomb targets in Drosophila melanogaster.
Polycomb group (PcG) complexes are multiprotein assemblages that bind to chromatin and establish chromatin states leading to epigenetic silencing. PcG proteins regulate homeotic genes in flies and vertebrates, but little is known about other PcG targets and the role of the PcG in development, differentiation and disease. Here, we determined the distribution of the PcG proteins PC, E(Z) and PSC and of trimethylation of histone H3 Lys27 (me3K27) in the D. melanogaster genome. At more than 200 PcG target genes, binding sites for the three PcG proteins colocalize to presumptive Polycomb response elements (PREs). In contrast, H3 me3K27 forms broad domains including the entire transcription unit and regulatory regions. PcG targets are highly enriched in genes encoding transcription factors, but they also include genes coding for receptors, signaling proteins, morphogens and regulators representing all major developmental pathways.


The redox properties of ascorbate peroxidase.
Reduction potentials for the catalytic compound I/compound II and compound II/Fe3+ redox couples, and for the two-electron compound I/Fe3+ redox couple, have been determined for ascorbate peroxidase (APX) and for a number of site-directed variants. For the wild type enzyme, the values are E degrees '(compound I/compound II) = 1156 mV, E degrees '(compound II/Fe3+) = 752 mV, and E degrees '(compound I/Fe3+) = 954 mV. For the variants, the analysis also includes determination of Fe3+/Fe2+ potentials which were used to calculate (experimentally inaccessible) E degrees '(compound II/Fe3+) potentials. The data provide a number of new insights into APX catalysis. The measured values for E degrees '(compound I/compound II) and E degrees '(compound II/Fe3+) for the wild type protein account for the much higher oxidative reactivity of compound I compared to compound II, and this correlation holds for a number of other active site and substrate binding variants of APX. The high reduction potential for compound I also accounts for the known thermodynamic instability of this intermediate, and it is proposed that this instability can account for the deviations from standard Michaelis kinetics observed for most APX enzymes during steady-state oxidation of ascorbate. This study provides the first systematic evaluation of the redox properties of any ascorbate peroxidase using a number of methods, and the data provide an experimental and theoretical framework for accurate determination of the redox properties of Fe3+, compound I, and compound II species in related enzymes.


Correlates of proximal premeditation among recently hospitalized suicide attempters.
Different conceptualizations of an impulsive suicide attempt (ISA) have not been studied systematically and there is no standard assessment of an ISA. This lack of clarity hinders the advancement of suicidological research and knowledge. The aim of the current study was to examine clinical correlates of different facets of an ISA (reduced proximal contemplation, planning, and decision to act) across divergent methodologies. Participants included 212 recent suicide attempters presenting to a Level 1 trauma hospital. The Suicide Intent Scale and the Timeline Follow-Back Interview for suicide attempts were used to assess different facets of an ISA and their associations with other attempt characteristics, and proximal and distal clinical correlates. A large percentage of patients had an ISA using facets of varying severity (ranging from 42% [contemplation] to 85% [decision]). Multivariate analyses revealed unique associations between a particular ISA facet and the following: hopelessness and depressive symptoms (contemplation), subjective expectation of fatality (planning), and acute negative life events (decision). Validated self-report screening measures were used to assess current psychopathology and future studies should include structured interviews to assess diagnostic features. Our results suggest that a fine-grained approach is needed for furthering our understanding of the ISA construct. In light of the current findings, ISA should be seriously considered given clinicians' task of determining whether a particular patient is at imminent risk for suicide.


The lone-pair-electron-driven phase transition and order-disorder processes in thermochromic (2-MIm)SbI4 organic-inorganic hybrid.
The easy to prepare and stable in air (2-methylimidazolium) tetraiodoantimoniate(iii) single-crystals with optical band gap of 2.17(1) eV at room temperature have been synthesized. The crystal structure features one-dimensional [SbI4]-n anionic chains, which are intercepted with stacks of 2MIm+ ions. At 294/295 K, it undergoes a structural phase transition to an incommensurately modulated phase as a result of subtle, lone-pair-electron-driven distortions of the anions. Separately from the anion displacements, the ordering of 2MIm+ countercations takes place over a wide temperature range of the modulated phase. The disorder changes from dynamic to static around 200 K, which affects the crystal structure leading to discontinuities and step-like contraction of the lattice parameters. The material is thermochromic with prominent color changes, from raspberry to yellow at low temperatures. The calculated electronic structures and observed optical properties signify its semiconducting character.


Targeting tumour hypoxia: shifting focus from oxygen supply to demand.
Tumour hypoxia is a well-recognised barrier to anti-cancer therapy and represents one of the best validated targets in oncology. Previous attempts to tackle hypoxia have focussed primarily on increasing tumour oxygen supply; however, clinical studies using this approach have yielded only modest clinical benefit, with often significant toxicity and practical limitations. Therefore, there are currently no anti-hypoxia treatments in widespread clinical use. As an emerging alternative strategy, we discuss the relevance of inhibiting tumour oxygen metabolism to alleviate hypoxia and highlight recently initiated clinical trials using this approach.


Asymptomatic Cardiac Ischemia Pilot (ACIP) Study. Relationship between exercise-induced and ambulatory ischemia in patients with stable coronary disease.
We investigated whether the presence and frequency of asymptomatic ischemic episodes recorded during ambulatory ECG (AECG) monitoring could be predicted on the basis of clinical characteristics or exercise treadmill test (ETT) performance in patients with stable coronary disease and whether the estimate of ischemia severity was similar between the AECG and ETT. Patients screened for the Asymptomatic Cardiac Ischemia Pilot (ACIP) study were selected for the current analysis if data were available from 48-hour AECG monitoring as well as from an ETT during which the patient developed > or = 1-mm ST-segment depression. Exercise ECG data were available for 143 of the 910 patients without ischemic episodes and for 659 of the 910 patients with ischemic episodes during AECG monitoring. Angina was more frequent among patients with ambulatory ischemic episodes than among patients without such ischemia (P < .001). Patients with AECG ischemia had a consistently more marked ischemic response on the ETT than patients without AECG ischemia; patients likely to have AECG ischemia could be predicted on the basis of ETT performance characteristics. However, the correlation coefficients between the severity of ischemia estimated by ETT and by AECG were small. There are significant relations between ischemia detected by AECG monitoring and by ETT, but the relations are limited, indicating that the two tests are not redundant to characterize coronary patients. A larger study investigating the prognostic significance of the ischemia identified by each modality, with follow-up for clinical events, will be necessary to determine the most appropriate methods to evaluate patients with stable coronary disease.


State-dependent mechanisms of LTP expression revealed by optical quantal analysis.
The expression mechanism of long-term potentiation (LTP) remains controversial. Here we combine electrophysiology and Ca(2+) imaging to examine the role of silent synapses in LTP expression. Induction of LTP fails to change p(r) at these synapses but instead mediates an unmasking process that is sensitive to the inhibition of postsynaptic membrane fusion. Once unmasked, however, further potentiation of formerly silent synapses leads to an increase in p(r). The state of the synapse thus determines how LTP is expressed.


Sediment quality in freshwater impoundments at Savannah National Wildlife Refuge.
Freshwater impoundments at Savannah National Wildlife Refuge (NWR), South Carolina, provide an important habitat for wildlife species, but degraded sediment quality in the Savannah River downstream of the discharge from two impoundments have caused concern about potential contaminant problems within the impoundments. The quality of sediments from five impoundments (impoundments no. 1, 2, 6, 7, and 17) on the NWR was evaluated using physical and chemical characterization, contaminant concentrations (metals, organochlorine pesticides, polychlorinated biphenyls, and polycyclic aromatic hydrocarbons), and toxicity testing. Survival of Hyalella azteca (freshwater amphipod) exposed for 28 days to solid-phase sediments was not significantly different from controls, but growth was significantly decreased at several sites. Survival in 96-hour exposures to sediment pore water was significantly decreased at most sites. Factors contributing to the toxic responses were low pH (3.7 to 4.1), ammonia (20 mg/L), and increased concentrations of cations in the pore water. The excess of simultaneously extracted metals over the acid volatile sulfides in the sediments was also typical of sites displaying decreased sediment quality. Elemental concentrations in pore water were negatively correlated with pH, and the highest concentrations were observed in impoundment no. 7. The acidic nature of the sediment in this impoundment was exacerbated by recent draining, burning, and disking, which allowed oxidation of the previously anoxic wetland sediment. Sediment disturbance and mixing of vegetation into the sediments by disking may also have contributed to the formation of ammonia caused by microbial decomposition of the fragmented organic matter. Contaminants were not detected in sediments from the impoundments, but releases of acidic water with increased levels of sediment cations from the impoundments may have contributed to the degraded sediment conditions previously observed in the river. The practice of dewatering sediments for vegetation control may exacerbate the acidification of vulnerable sediments within impoundments of this NWR.


Synaptic shot noise and conductance fluctuations affect the membrane voltage with equal significance.
The subthreshold membrane voltage of a neuron in active cortical tissue is a fluctuating quantity with a distribution that reflects the firing statistics of the presynaptic population. It was recently found that conductance-based synaptic drive can lead to distributions with a significant skew. Here it is demonstrated that the underlying shot noise caused by Poissonian spike arrival also skews the membrane distribution, but in the opposite sense. Using a perturbative method, we analyze the effects of shot noise on the distribution of synaptic conductances and calculate the consequent voltage distribution. To first order in the perturbation theory, the voltage distribution is a gaussian modulated by a prefactor that captures the skew. The gaussian component is identical to distributions derived using current-based models with an effective membrane time constant. The well-known effective-time-constant approximation can therefore be identified as the leading-order solution to the full conductance-based model. The higher-order modulatory prefactor containing the skew comprises terms due to both shot noise and conductance fluctuations. The diffusion approximation misses these shot-noise effects implying that analytical approaches such as the Fokker-Planck equation or simulation with filtered white noise cannot be used to improve on the gaussian approximation. It is further demonstrated that quantities used for fitting theory to experiment, such as the voltage mean and variance, are robust against these non-Gaussian effects. The effective-time-constant approximation is therefore relevant to experiment and provides a simple analytic base on which other pertinent biological details may be added.


Picoliter droplet formation on thin optical fiber tips.
In this paper, we present experimental results on how minute droplets are formed on fiber optic end faces. Results show that reproducible picoliter volumes can be generated when fibers are retracted from an aqueous phase contained under an inert fluorinated immiscible liquid, with a coefficient of variation (CV) of 0.7-2.3%. The droplet formation was analyzed as a function of the fiber diameter, retraction speed, and wettability. Experiments reveal a volume-determining critical equilibrium contact angle between 60 degrees and 75 degrees , defining the onset of fiber end-face dewetting. The dynamics of the droplet snap-off progression was characterized using high-speed imaging in order to explain the observed wettability-volume dependency.


Relapses in patients with a systemic vasculitis.
Treatment with cyclophosphamide and steroids has greatly improved survival in patients with systemic necrotizing vasculitis but does not always provide a complete cure. There are as yet few data on the incidence, pattern and outcome of relapses in these diseases. We studied relapses in 150 consecutive patients with an idiopathic necrotizing vasculitis: 12 with classical polyarteritis (CPAN); 95 with microscopic polyarteritis (MPA); 28 with Wegener's granulomatosis (WG); and 15 with limited Wegener's granulomatosis (LWG). The relapse rates and median time to relapse in months were: CPAN, 41.7%/33; MPA, 25.4%/24; WG, 44%/42; LWG, 52%/18. The clinical features of relapse were similar to or more aggressive than those of the original presentation in CPAN and LWG and included renal disease for the first time, but in MPA and WG, relapse involved less renal involvement in the majority of cases. Laboratory tests, although often positive at relapse, were unhelpful in its prediction. The considerable accumulative non-fatal relapse rate contrasts with the very good long-term survival rates, and confirms the importance of long-term follow-up in systemic vasculitis.


Endogenous viruses: Connecting recent and ancient viral evolution.
The rapid rates of viral evolution allow us to reconstruct the recent history of viruses in great detail. This feature, however, also results in rapid erosion of evolutionary signal within viral molecular data, impeding studies of their deep history. Thus, the further back in time, the less accurate the inference becomes. Furthermore, reconstructing complex histories of transmission can be challenging, especially where extinct viral lineages are concerned. This problem has been partially solved by the discovery of viruses embedded in host genomes, known as endogenous viral elements (EVEs). Some of these endogenous viruses are derived from ancient relatives of extant viruses, allowing us to better examine ancient viral host range, geographical distribution and transmission routes. Moreover, our knowledge of viral evolutionary timescales and rate dynamics has also been greatly improved by their discovery, thereby bridging the gap between recent and ancient viral evolution.


Electrophysiological heterogeneity of spinally projecting serotonergic and nonserotonergic neurons in the rostral ventromedial medulla.
This study examined the passive membrane and action potential properties of serotonergic and nonserotonergic neurons in the rostral ventromedial medulla (RVM) of the rat using whole cell patch-clamp recording techniques in the slice. Serotonergic neurons were identified by immunoreactivity for tryptophan hydroxylase (TrpH). Spinally projecting neurons were retrogradely labeled with 1'-dioactadecyl-3,3,3',3'-tetramethylindocarbodyanine perchlorate (DiI). Three types of neurons were identified within both spinally projecting serotonergic and nonserotonergic populations. Type 1 neurons exhibited irregular or sporadic spontaneous activity interspersed with periods of quiescence. Type 2 neurons were not spontaneously active and were additionally discriminated by a more negative resting membrane potential and a larger-amplitude action potential. Type 3 neurons fired repetitively without pause. Serotonergic neurons had a higher membrane resistance and greater action potential half-width than their nonserotonergic counterparts and rarely exhibited a fast afterhyperpolarization. Serotonergic type 3 neurons also fired more slowly and regularly than nonserotonergic type 3 neurons. Comparison of electrophysiological and immunohistochemical characteristics suggested that the smallest type 3 serotonergic neurons had an increased risk of immunohistochemical "misclassification" due to failure to detect TrpH, possibly due to more complete dialysis of intracellular contents during lengthy recordings. This risk was minimal for type 1 or 2 serotonergic neurons. The three different types of spinally projecting serotonergic neurons also differed markedly in their responsiveness to the mu opioid receptor agonist D-Ala2, NMePhe4, Gly5-ol]enkephalin. These results provide important new electrophysiological and pharmacological evidence for a significant heterogeneity among spinally projecting serotonergic RVM neurons. They may also provide a basis for resolving the controversy concerning the role of serotonergic RVM neurons in opioid analgesia.


Factors obstetricians, family physicians and midwives consider when counselling women about a trial of labour after caesarean and planned repeat caesarean: a qualitative descriptive study.
Repeat caesarean sections (CSs) are major contributors to the high rate of CS in Canada and globally. Women's decisions to have a planned repeat CS (PRCS) or a trial of labour after CS (TOLAC) are influenced by their maternity care providers. This study explored factors maternity care providers consider when counselling pregnant women with a previous CS, eligible for a TOLAC, about delivery method. A qualitative descriptive design was implemented. Semi-structured, one-to-one in-depth telephone interviews were conducted with 39 maternity care providers in Ontario, Canada. Participants were recruited at 2 maternity care conferences and with the use of snowball sampling. Interviews were audio recorded and transcribed verbatim. Data were uploaded into the data management software, NVIVO 10.0 and analyzed using qualitative content analysis. Participants consisted of 12 obstetricians, 13 family physicians and 14 midwives. Emergent themes, reflecting the factors maternity care providers considered when counselling on mode of delivery, were organized under the categories clinical/patient factors, health system factors and provider preferences. Maternity care providers considered clinical/patient factors, including women's choice … with conditions, their assessment of women's chances of a successful TOLAC, their perception of women's risk tolerance, women's preferred delivery method, and their perception of women's beliefs and attitudes about childbirth. Additionally, providers considered health system factors which included colleague support for TOLAC and time needed to mount an emergency CS. Finally, provider factors emerged as considerations when counselling. They included provider preference for PRCS or TOLAC, provider scope of practice, financial incentives and convenience related to PRCS, past experiences with TOLAC and PRCS and providers' perspectives on risk of TOLAC. The findings highlight the multiplicity of factors maternity care providers consider when counselling women. Effectively addressing clinical, health care system and personal factors that influence counselling may help decrease non-medically indicated PRCS.


A computationally efficient method for automatic registration of orthogonal x-ray images with volumetric CT data.
The paper presents a computationally efficient 3D-2D image registration algorithm for automatic pre-treatment validation in radiotherapy. The novel aspects of the algorithm include (a) a hybrid cost function based on partial digitally reconstructed radiographs (DRRs) generated along projected anatomical contours and a level set term for similarity measurement; and (b) a fast search method based on parabola fitting and sensitivity-based search order. Using CT and orthogonal x-ray images from a skull and a pelvis phantom, the proposed algorithm is compared with the conventional ray-casting full DRR based registration method. Not only is the algorithm shown to be computationally more efficient with registration time being reduced by a factor of 8, but also the algorithm is shown to offer 50% higher capture range allowing the initial patient displacement up to 15 mm (measured by mean target registration error). For the simulated data, high registration accuracy with average errors of 0.53 mm +/- 0.12 mm for translation and 0.61 +/- 0.29 degrees for rotation within the capture range has been achieved. For the tested phantom data, the algorithm has also shown to be robust without being affected by artificial markers in the image.


Hormonal regulation of fruitlet abscission induced by carbohydrate shortage in citrus.
The hormonal signals controlling fruitlet abscission induced by sugar shortage in citrus were identified in Satsuma mandarin, Citrus unshiu (Mak.) Marc, cv. Clausellina and cv. Okitsu. Sugar supply, hormonal responses and fruitlet abscission were manipulated through full, partial or selective leaf removals at anthesis and thereafter. In developing fruitlets, defoliations reduced soluble sugars (up to 98%), but did not induce nitrogen and water deficiencies. Defoliation-induced abscission was preceded by rises (up to 20-fold) in the levels of abscisic acid (ABA) and 1-aminocyclopropane-1-carboxylic acid (ACC) in fruitlets. Applications to defoliated plants showed that ABA increased ACC levels (2-fold) and accelerated fruitlet abscission, whereas norflurazon and 2-aminoethoxyvinyl glycine reduced ACC (up to 65%) and fruitlet abscission (up to 40%). Only the full defoliation treatment reduced endogenous gibberellin A1 (4-fold), whereas exogenous gibberellins had no effect on abscission. The data indicate that fruitlet abscission induced by carbon shortage in citrus is regulated by ABA and ACC originating in the fruits, while gibberellins are apparently implicated in the maintenance of growth. In this system, ABA may act as a sensor of the intensity of the nutrient shortage that modulates the levels of ACC and ethylene, the activator of abscission. This proposal identifies ABA and ACC as components of the self-regulatory mechanism that adjusts fruit load to carbon supply, and offers a physiological basis for the photoassimilate competition-induced abscission occurring under natural conditions.


Phosphorylation of the F(1)F(o) ATP synthase beta subunit: functional and structural consequences assessed in a model system.
We previously discovered several phosphorylations to the beta subunit of the mitochondrial F(1)F(o) ATP synthase complex in isolated rabbit myocytes on adenosine treatment, an agent that induces cardioprotection. The role of these phosphorylations is unknown. The present study focuses on the functional consequences of phosphorylation of the ATP synthase complex beta subunit by generating nonphosphorylatable and phosphomimetic analogs in a model system, Saccharomyces cerevisiae. The 4 amino acid residues with homology in yeast (T58, S213, T262, and T318) were studied with respect to growth, complex and supercomplex formation, and enzymatic activity (ATPase rate). The most striking mutant was the T262 site, for which the phosphomimetic (T262E) abolished activity, whereas the nonphosphorylatable strain (T262A) had an ATPase rate equivalent to wild type. Although T262E, like all of the beta subunit mutants, was able to form the intact complex (F(1)F(o)), this strain lacked a free F(1) component found in wild-type and had a corresponding increase of lower-molecular-weight forms of the protein, indicating an assembly/stability defect. In addition, the ATPase activity was reduced but not abolished with the phosphomimetic mutation at T58, a site that altered the formation/maintenance of dimers of the F(1)F(o) ATP synthase complex. Taken together, these data show that pseudophosphorylation of specific amino acid residues can have separate and distinctive effects on the F(1)F(o) ATP synthase complex, suggesting the possibility that several of the phosphorylations observed in the rabbit heart can have structural and functional consequences to the F(1)F(o) ATP synthase complex.


Role of energy systems in two intermittent field tests in women field hockey players.
The energetics of 2 field tests that reflect physical performance in intermittent sports (i.e., the Interval Shuttle Sprint Test [ISST] and the Interval Shuttle Run Test [ISRT]) were examined in 21 women field hockey players. The ISST required the players to perform 10 shuttle sprints starting every 20 seconds. During the ISRT, players alternately ran 20-m shuttles for 30 seconds and walked for 15 seconds with increasing speed. Anaerobic and aerobic power tests included Wingate cycle sprints and a .V(O2)max cycle test, respectively. Based on correlation and regression analyses, it was concluded that for the ISST, anaerobic energetic pathways contribute mainly to energy supply for peak sprint time, while aerobic energetic pathways also contribute to energy supply for total sprint time. Energy during the ISRT is supplied mainly by the aerobic energy system. Depending on the aspect of physical performance a coach wants to determine, the ISST or ISRT can be used.


Increased HIV testing will modestly reduce HIV incidence among gay men in NSW and would be acceptable if HIV testing becomes convenient.
Determine the acceptability and epidemiological impact of increases in HIV testing in gay men in New South Wales (NSW), Australia- particularly pertinent when considering treatment as prevention and the need to reduce undiagnosed infections. We conducted an online survey and focus groups to assess whether increases in HIV testing would be acceptable to gay men in NSW. In parallel, we assessed the potential impact of increases in testing coverage and/or frequency using an individual-based model of HIV transmission. If sexual practices and the rate of initiating HIV treatment are unchanged then increasing HIV testing reduces infections. Increasing testing frequency has the largest impact, with a 13.8% reduction in HIV infections over 10 years if the ∼55-75% of men who test at least once per year increased their testing frequency to four times per year. If testing levels decrease from current levels then we expect an increase in HIV infections with a sharply rising trend over time. Increasing HIV testing would be acceptable if testing was more convenient. However, only ∼25% of men surveyed were 'very likely' to increase their level of HIV testing. Men delayed or avoided testing due to the slowness in obtaining results and if they believed they had not put themselves at risk. An increase in HIV testing alone is unlikely to reduce HIV incidence substantially in NSW gay men- however, the relatively high testing levels need to continue to prevent an increase in HIV infections. In jurisdictions with lower levels of HIV testing, increases in testing coverage and frequency are likely to have a larger impact. Successful treatment as prevention interventions will require increases in testing rates; such increases would be acceptable to gay men in NSW but only if more convenient testing and rapid communication of results were available.


Effects of Helicobacter hepaticus on the proteome of HEp-2 cells.
Helicobacter hepaticus infects the bowel and biliary tree of several animals, producing inflammation. Colonisation of mouse livers can induce hepatocellular carcinomas. The effects of H. hepaticus on the proliferation and global protein expression of human HEp-2 cells were studied by examining the changes in the protein profiles of cells exposed to the bacterium. HEp-2 cells were grown for four days under a microaerobic atmosphere or under the same conditions in co-cultures with H. hepaticus at various inoculum densities. Enlargement, distension and elongation of HEp-2 cells were observed in co-cultures with H. hepaticus. The number of live cells declined by only an order of magnitude at bacterial inocula of approximately 10(9)cfu/ml, but were reduced to less than 10(3)cells/ml at approximately 10(10)cfu/ml bacteria inocula. Protein expression by HEp-2 cells was investigated employing two-dimensional gel electrophoresis. In cells grown with or without bacteria, 17 differentially expressed proteins were identified by tandem mass spectrometry. These proteins participated in several biological functions including amino acid metabolism, cell growth and proliferation, stress response, protein translation and modification, etc. The onset of a catastrophic killing of HEp-2 cells at a bacterial density of approximately 10(9)cfu/ml suggested a multimodal action for H. hepaticus infection, and the modulation of the expression of proteins involved in different biological functions showed that the presence of H. hepaticus has broad effects on the physiology of HEp-2 cells.


Structural characterization of HPM-7, a more ordered than expected germanosilicate zeolite.
HPM-7, a germanosilicate synthesized using long imidazolium-based dications with two different linkers, is shown to possess the POS topology, although disorder may exist but it is very difficult to discern. First, three simple ordered polymorphs (POS-A to POS-C) with very similar energies and structural motifs could give rise to intergrowths that would be very difficult to recognize by powder X-ray diffraction, according to DIFFaX simulations. Another four structures (POS-D to POS-G) can be derived from POS by changing the orientation of two single four rings within the structure, possibly providing an additional source of disorder. While 3D EDT strongly suggests that HPM-7 basically possesses the POS-A (i.e. POS) topology, a detailed HR-STEM study demonstrates the rare existence of some disorder compatible with the polymorph POS-D. The general avoidance of polymorphs with very similar structural motifs and comparable energies points to a rather specific structure-direction by the organic dications used.


A glycomimetic compound inhibits DC-SIGN-mediated HIV infection in cellular and cervical explant models.
Dendritic cell-specific intercellular adhesion molecule (ICAM)-3 grabbing nonintegrin (DC-SIGN) participates in the initial stages of sexually transmitted HIV-1 infection by recognizing highly mannosylated structures presented in multiple copies on HIV-1 gp120 and promoting virus dissemination. Inhibition of HIV interaction with DC-SIGN thus represents a potential therapeutic approach for viral entry inhibition at the mucosal level. Herein we evaluate the efficacy in inhibiting HIV-1 infection and the potential toxicity of a multimeric glycomimetic DC-SIGN ligand (Dendron 12). The ability of Dendron 12 to block HIV-1 infection was assessed in cellular and human cervical explant models. Selectivity of Dendron 12 towards DC-SIGN and langerin was evaluated by surface plasmon resonance studies. β chemokine production following stimulation with Dendron 12 was also analyzed. Toxicity of the compound was evaluated in cellular and tissue models. Dendron 12 averted HIV-1 trans infection of CD4(+) T lymphocytes in presence of elevated viral loads and prevented HIV-1 infection of human cervical tissues, under conditions mimicking compromised epithelial integrity, by multiple clades of R5 and X4 tropic viruses. Treatment with Dendron 12 did not interfere with the activity of langerin and also significantly elicited the production of the β chemokines MIP-1α, MIP-1β and RANTES. Dendron 12 thus inhibits HIV-1 infection by competition with binding of HIV to DC-SIGN and stimulation of β-chemokine production. Dendron 12 represents a promising lead compound for the development of anti-HIV topical microbicides.


Implementation of a workflow system with electronic verification for preparation of oral syringes.
The implementation of a commercially available, Web-based, barcode-enabled workflow management system for filling and checking patient-specific oral syringes at the Cleveland Clinic is described. Historically, the process for preparing oral syringes has been a manual, repetitive procedure at Cleveland Clinic. Within this process, paper logs are the sole source of information. These logs can be difficult to locate or interpret. It remains a rote process with numerous manual steps offering opportunity for human error. Traditionally, automation, such as barcode scanning, has not been available during oral syringe preparation and checking. Based on increasing regulatory demands and documentation gaps identified, solutions were explored in the pharmacy's nonsterile compounding environment. An electronic verification workflow system was developed, implemented, and evaluated with a focused assessment of throughput and patient safety relative to oral syringe preparation. This was a retrospective study conducted in an academic, tertiary, acute care medical center. Analysis was completed at 3 months to evaluate efficiencies. Data regarding experiences at the Cleveland Clinic shows promising benefits with regards to United States Pharmacopeia chapter 795 compliance, the pursuit of incorporating advanced informatics and automation in manual processes, and standardization of workflow within an enterprise setting. Implementation of an electronic workflow verification system that integrates barcode verification and image-capture capabilities has maintained regulatory compliance in the nonsterile compounding environment including preparation of oral syringe doses without impacting throughput.


Genetic and intermediate phenotypic susceptibility markers of gastric cancer in Hispanic Americans: a case-control study.
Hispanics are the largest nonwhite ethnic group in the US population, and they have higher incidence and mortality rates for gastric cancer (GC) than whites and Asians. Studies have identified several genetic susceptibility loci and intermediate phenotypic biomarkers for GC in whites and Asians. No studies have evaluated genetic susceptibility and intermediate phenotypic biomarkers in Hispanics. In a case-control study of 132 Hispanic patients with GC (cases) and a control group of 125 Hispanics (controls), the authors evaluated the association of 5 single nucleotide polymorphisms (SNPs) that predispose whites and/or Asians to GC and of 2 intermediate phenotypic markers in peripheral blood leukocytes, ie, telomere length and mitochondrial DNA (mtDNA) copy number, with the GC risk. The variant C allele of the reference SNP rs2294008 in the PSCA gene was associated with a significantly reduced risk of GC (per allele-adjusted odds ratio [aOR], 0.51; 95% confidence interval [CI], 0.33-0.77; P = .002). Leukocyte mtDNA copy numbers were significantly lower in GC cases (mean ± standard deviation, 0.91 ± 0.28) than in controls (1.29 ± 0.42; P < .001). When individuals were dichotomized into high and low mtDNA copy number groups based on the median mtDNA copy number value in the controls, those who had a low mtDNA copy number had a significantly increased risk of GC (aOR, 11.00; 95% CI, 4.79-25.23; P < .001) compared with those who had a high mtDNA copy number. Telomere length was not associated significantly with the risk of GC (aOR, 1.21; 95% CI, 0.65-2.27; P = .551). Hispanics share certain genetic susceptibility loci and intermediate phenotypic GC biomarkers with whites and Asians and may also have distinct genetic susceptibility factors.


Sleep-onset insomniacs have delayed temperature rhythms.
It was predicted from free running and ultradian cycle studies that sleep-onset insomniacs would have endogenous circadian rhythms that were phase delayed compared to good sleepers. Thirteen sleep-onset insomniacs and nine good sleepers were selected to differ only in their sleep-onset latencies as confirmed by polysomnography. their rectal temperatures were measured over a 26-h constant routine and analyzed with best-fit Fourier curves including 24-h fundamental and 12-h harmonic components. The temperature rhythm markers of the insomniacs' rhythms were approximately 2.5 h later than the respective phases of the good sleepers. The usual bedtimes of the insomniacs fell within the "wake maintenance zone" of their delayed temperature rhythm. The good sleepers had typical bedtimes several hours after their "wake maintenance zone" and closer to their body temperature minimum. It was suggested that manipulations to phase advance the insomniacs' rhythms would reduce their sleep-onset latencies. It was also predicted that early morning insomnia results from phase advanced circadian rhythms and that sleep maintenance insomnia results from an abnormal phase relationship between the 24-h temperature rhythm and 12-h sleep-alert rhythm.