Exploring the roles of Mus81-Eme1/Mms4 at perturbed replication forks.
Cells of all living organisms have evolved complex mechanisms that serve to stabilise, repair and restart stalled, blocked and broken replication forks. The heterodimeric Mus81-Eme1/Mms4 structure-specific endonuclease appears to play an important role(s) in homologous recombination-mediated processing of such perturbed forks. This enzyme has been implicated in the cleavage of stalled and blocked replication forks to initiate recombination, as well as in the processing of recombination intermediates that result from repairing damaged forks. In this review we assess the biochemical and genetic evidence for the mitotic role of Mus81-Eme1/Mms4 at replication forks and in repairing post-replication DNA damage. Mus81 appears to act when replication is impeded by genotoxins or by impairment of the replication machinery, or when arrested replication forks are not adequately protected. We discuss how its action is regulated by the S-phase cell cycle checkpoint, depending on the nature of the stalled or damaged fork. We also present a new way in which Mus81 may limit crossing over during the repair of post-replication gaps, and explore Mus81's interplay with other components of the recombination machinery, including the RecQ helicases that also play important roles in processing replication and recombination intermediates.


Effectiveness of weight management interventions in children: a targeted systematic review for the USPSTF.
Targeted systematic review to support the updated US Preventive Services Task Force (USPSTF) recommendation on screening for obesity in children and adolescents. To examine the benefits and harms of behavioral and pharmacologic weight-management interventions for overweight and obese children and adolescents. Our data sources were Ovid Medline, PsycINFO, the Education Resources Information Center, the Database of Abstracts of Reviews of Effects, the Cochrane databases, reference lists of other reviews and trials, and expert recommendations. After 2 investigators reviewed 2786 abstracts and 369 articles against inclusion/exclusion criteria, we included 15 fair- to good-quality trials in which the effects of treatment on weight, weight-related comorbidities, and harms were evaluated. Studies were quality rated by 2 investigators using established criteria. Investigators abstracted data into standard evidence tables. In the available research, obese (or overweight) children and adolescents aged 4 to 18 years were enrolled, and no studies targeted those younger than 4 years. Comprehensive behavioral interventions of medium-to-high intensity were the most effective behavioral approach with 1.9 to 3.3 kg/m(2) difference favoring intervention groups at 12 months. More limited evidence suggests that these improvements can be maintained over the 12 months after the end of treatments and that there are few harms with behavioral interventions. Two medications combined with behavioral interventions resulted in small (0.85 kg/m(2) for orlistat) or moderate (2.6 kg/m(2) for sibutramine) BMI reduction in obese adolescents on active medication; however, no studies followed weight changes after medication use ended. Potential adverse effects were greater than for behavioral interventions alone and varied in severity. Only 1 medication (orlistat) has been approved by the US Food and Drug Administration for prescription use in those aged > or =12 years. Over the past several years, research into weight management in obese children and adolescents has improved in quality and quantity. Despite important gaps, available research supports at least short-term benefits of comprehensive medium- to high-intensity behavioral interventions in obese children and adolescents.


Characteristics and prognosis of Japanese female breast cancer patients: The BioBank Japan project.
Breast cancer is currently the most common type of cancer in Japanese females. Unlike most other types of cancer, breast cancer develops more frequently in middle-aged females than in elderly females. Of all Japanese female breast cancer patients aged ≥20 years whom the BioBank Japan Project originally enrolled between 2003 and 2008, 2034 were registered within 90 days after their diagnosis. We described the lifestyle and clinical characteristics of these patients at study entry. Furthermore, we examined the effect of these characteristics on all-cause mortality. In the female patients registered within 90 days after diagnosis, the frequency of stage 0 or unclassified, stage I, II, III and IV were 11.4%, 47.9%, 37.0%, 2.9% and 0.8%, respectively. The proportion of histological types was 12.9% for non-invasive carcinoma (ductal carcinoma and lobular carcinoma), 81.0% for invasive carcinoma (papillotubular carcinoma, solid tubular carcinoma, scirrhous carcinoma and special types), 0.2% for Paget's diseases and 5.8% for others. Those positive for the estrogen and progesterone receptors accounted for 75.8% and 62.1% of all patients, respectively. Among 1860 female participants registered within 90 days, 218 participants died during 144,54 person-years of follow-up. More advanced stage, elevation of serum carcinoembryonic antigen and carbohydrate antigen 15-3 levels and absence of the estrogen receptor at study entry were crudely associated with an increased risk of all-cause mortality after adjustment for age. This study showed the association of several clinical characteristics with all-cause mortality in female breast cancer patients.


Anatomy of extrahepatic portosystemic shunts in dogs as determined by computed tomography angiography.
Congenital extrahepatic portosystemic shunts are anomalous vessels joining portal and systemic venous circulation. These shunts are often diagnosed sonographically, but computed tomography (CT) angiography produces high-resolution images that give a more comprehensive overview of the abnormal portal anatomy. CT angiography was performed on 25 dogs subsequently proven to have an extrahepatic portosystemic shunt. The anatomy of each shunt and portal tributary vessels was assessed. Three-dimensional images of each shunt type were created to aid understanding of shunt morphology. Maximal diameter of the extrahepatic portosystemic shunt and portal vein cranial and caudal to shunt origin was measured. Six general shunt types were identified: splenocaval, splenoazygos, splenophrenic, right gastric-caval, right gastric-caval with a caudal shunt loop, and right gastric-azygos with a caudal shunt loop. Slight variations of tributary vessels were seen within some shunt classes, but were likely clinically insignificant. Two shunt types had large anastomosing loops whose identification would be important if surgical correction were attempted. A portal vein could not be identified cranial to the shunt origin in two dogs. In conclusion, CT angiography provides an excellent overview of extrahepatic portosystemic shunt anatomy, including small tributary vessels and loops. With minor variations, most canine extrahepatic portosystemic shunts will likely be one of six general morphologies.


The emerging role of electronic medical records in pharmacogenomics.
Health-care information technology and genotyping technology are both advancing rapidly, creating new opportunities for medical and scientific discovery. The convergence of these two technologies is now facilitating genetic association studies of unprecedented size within the context of routine clinical care. As a result, the medical community will soon be presented with a number of novel opportunities to bring functional genomics to the bedside in the area of pharmacotherapy. By linking biological material to comprehensive medical records, large multi-institutional biobanks are now poised to advance the field of pharmacogenomics through three distinct mechanisms: (i) retrospective assessment of previously known findings in a clinical practice-based setting, (ii) discovery of new associations in huge observational cohorts, and (iii) prospective application in a setting capable of providing real-time decision support. This review explores each of these translational mechanisms within a historical framework.


Anticariogenic effects of green tea.
The effects of green tea extract on caries inhibition of hamsters and on acid resistance of human tooth enamel have been investigated. Both in vivo and in vitro experiments showed that original extract of green tea had the significant effects on these points. The dialyzed tea solution in which the fluoride was removed almost completely, also showed the remarkable effects both in vivo and in vitro experiment as is similar to the original tea extract. The results obtained from this study suggested that fluoride in green tea may play a role to increase the cariostatic action in cooperation with other components in tea. However, the action of fluoride does not seem to be so important, because its concentration is very low. The effect of green tea on caries inhibition as well as on the increment of acid resistance appears to be more correlative with the non-dialysable substances in tea.


Regulation of neurite outgrowth and SNAP-25 gene expression by the Brn-3a transcription factor.
SNAP-25 is a presynaptic nerve terminal protein which is also essential for the process of neurite outgrowth in vivo and in vitro. However the processes regulating its expression have not been characterized previously. We show that the gene encoding this protein, SNAP, is strongly activated by the Brn-3a POU (Pit-Oct-Unc) family transcription factor. Expression of both Brn-3a and SNAP-25 increases when ND7 neuronal cells are induced to extend neurite processes by serum removal. Inhibition of Brn-3a expression in these cells inhibits SNAP-25 expression and abolishes the neurite outgrowth that normally occurs in response to serum removal. These results identify Brn-3a as the first transcription factor having a role in process outgrowth in neuronal cells acting, at least in part, via the activation of SNAP-25 gene expression.


Challenges of a community based pragmatic, randomised controlled trial of weight loss maintenance.
Randomised controlled trials (RCTs) have a reputation for being inherently difficult to deliver as planned and often face unforeseen challenges and delays, particularly in relation to organisational and governance difficulties, participant interest, constraints due to allocation of costs, local investigator interest and lengthy bureaucracy. Recruitment is often difficult and the challenges faced often impact on the cost and delivery of a successful trial within the funded period. This paper reflects upon the challenges faced in delivering a pragmatic RCT of weight loss maintenance in a community setting and suggests some potential solutions. The weight loss maintenance in adults trial aimed to evaluate the impact of a 12 month, individually tailored weight maintenance intervention on BMI 3 years from randomisation. Participants were recruited primarily from participant identification centres (PICs)-GP surgeries, exercise on referral schemes and slimming world. The intervention was delivered in community settings. A recruitment strategy implementation plan was drafted to address and monitor poor recruitment. Delays in opening and recruitment were experienced early on. Some were beyond the control of the study team such as; disagreement over allocation of national health service costs and PIC classification as well as difficulties in securing support from research networks. That the intervention was delivered in community settings was often at the root of these issues. Key items to address at the design stage of future trials include feasibility of eligibility criteria. The most effective element of the recruitment implementation plan was to refocus sources of recruitment and target only those who could fulfil the eligibility criteria immediately. Learnings from this trial should be kept in mind by those designing similar studies in the future. Considering potential governance, cost and research network support implications at the design stage of pragmatic trials of any community-based complex intervention is paramount. The appropriateness and viability of inclusion criteria also require careful consideration as does use of a targeted advertising strategy. ISRCTN35774128, 12/01/2010.


Reflex control of facial flushing during body heating in man.
Facial temperature and amplitude of capillary pulsations from the forehead and cheeks were measured shortly before and after pharmacological blockade of the stellate ganglion in 9 patients with reflex sympathetic dystrophy of an upper limb, and in 1 other patient with erythromelalgia of all four limbs. Patients were then heated to determine the effect of sympathetic blockade on mediation of thermoregulatory facial flushing. Release of vasoconstrictor tone following stellate ganglion blockade was invariably followed by an increase in orbital and cheek temperature, and by similar but less consistent increases in temperature of the forehead, lips and chin. The amplitude of capillary pulsations recorded from the forehead and cheeks also increased on the side of stellate ganglion blockade. Flushing of the forehead and cheek on the sympathetically intact side during body heating far outweighed the extent of flushing after release of vasoconstrictor tone. The thermoregulatory response was prevented by sympathetic blockade, indicating that an active sympathetic vasodilator pathway had been interrupted. In contrast, the temperature of the orbit during body heating was not influenced by sympathetic blockade. Asymmetry of forehead temperature was detected before sympathetic blockade in 8 of 9 patients with reflex sympathetic dystrophy, suggesting that autonomic disturbances in this condition may influence cervical sympathetic outflow. The results indicate that sympathetic vasodilator fibres passing through the stellate ganglion mediate thermoregulatory facial flushing, and that release of vasoconstrictor tone has only a minor influence on this response in most areas of the face.


Complement inhibitor of C5 activation from the soft tick Ornithodoros moubata.
Blood-feeding ticks must control C activation or be damaged by the host inflammatory response. We report the characterization and expression of a novel, relatively small, broad-acting C inhibitory protein (termed OmCI) from the soft tick Ornithodoros moubata. The native 17-kDa nonglycosylated protein inhibits both human and guinea pig classical and alternative C activation pathways. The IC50 values for each pathway were 12 and 27 nM, respectively, in hemolytic assays using human serum diluted 40-fold. The cDNA encodes a protein of 168 aa, including an 18-aa secretion signal sequence that is absent in the mature form. The inhibitor has 46% amino acid identity with moubatin, a platelet aggregation inhibitor also from O. moubata that is an outlying member of the lipocalin family. Native OmCI had no inhibitory effect on the addition of C8 and C9 to preformed C5b-C7 and C5b-C8 to form the membrane attack complex and no effect on the rate of C3a production by the C3 convertase enzymes C4bC2a, C3(H2O)Bb, or C3bBb. Both recombinant and native OmCI abolish production of C5a by human classical (C4bC3bC2a) and alternative (C3bC3bBb) C5 convertases. Addition of excess C5 but not C3 competes away the inhibitory activity of OmCI, indicating that OmCI targets C5 itself rather than inhibiting the C5 convertase C4bC3bC2a itself. Direct binding of OmCI to C5 was demonstrated by Western blotting and gel filtration chromatography using 125I-labeled proteins. OmCI is the first lipocalin family member shown to inhibit C and also the first natural inhibitor that specifically targets the C5 activation step.


Knowledge to practice in developmental coordination disorder: utility of an evidence-based online module for physical therapists.
Developmental coordination disorder (DCD) is a chronic condition with potential negative health consequences. Therapists working with children with DCD need access to tailored, synthesized, evidence-based information; however a knowledge-to-practice gap exists. The aim of this study was to develop and evaluate the utility of an evidence-based online DCD module tailored to physical therapists' (PTs) needs. Guided by the Knowledge to Action framework, we interviewed PTs working with children with DCD (n = 9) to identify their information needs. Their recommendations, along with synthesized DCD research evidence, informed module development. These PTs as well as others (n = 50) evaluated the module's usefulness. The module incorporated important content areas including: (1) identification; (2) planning interventions and goals; (3) evidence-based practice; (4) management; and (5) resources. Case scenarios, clinical applications, interactive media, links to resources, and interactive learning opportunities were also embedded. PTs perceived the module to be comprehensive and useful and provided feedback to improve module navigation. Involving end-users throughout the development and evaluation of an online PT DCD module contributed to its relevance, applicability, and utility. It will be important to evaluate whether use of this module improves the quality of services provided by PTs.


The National Psychiatric Morbidity surveys of Great Britain--initial findings from the household survey.
This paper describes the Household Survey from the National Survey of Psychiatric Morbidity. This covered a sample drawn at random from the population of Britain, with the exception of the Highlands and Islands of Scotland. The Postcode Address file was used as the sampling frame. Nearly 13000 adults aged 16-65 were selected for interview, of whom 10108 (79.4%) were successfully interviewed. Eight per cent could not be contacted and 13% refused interview. Psychiatric assessment was carried out by lay interviewers using the CIS-R. Subjects were also screened for psychosis, and screen-positive individuals were examined by psychiatrists using SCAN. Sixteen per cent of subjects scored above the standard cut-off of 12 on the CIS-R. The overall 1-week prevalence of neurotic disorder was 12.3% in males and 19.5% in females. Unmarried and post-marital groups had high rates of disorder, as did single parents and people living on their own. Respondents in Social Class I had notably lower rates of neurotic disorder than the remainder of the sample. Unemployment was strongly associated with disorder. Subjects living in urban areas had a higher overall prevalence, but there was no significant variation by region. Black respondents had higher rates of disorders that were entirely explained by their age, family type and social class. Individual neurotic disorders were all significantly commoner in women, with the exception of panic disorder. The 1-year prevalence of functional psychoses was 4 per 1000, with no sex difference. Alcohol and drug dependence was considerably more prevalent in men. For the first time, the survey provides data on the prevalence and correlates of psychiatric disorder on a nationwide sample that can be used to inform equitable and effective national psychiatric services.


7201 carcinoids: increasing incidence overall and disproportionate mortality in the elderly.
The aim of the study was to determine outcomes for respiratory and gastrointestinal carcinoid tumors utilizing a large cancer registry. Cases of respiratory and gastrointestinal carcinoid from the Florida Cancer Data System (FCDS) from 1981 to 2001 were reviewed. Descriptive statistics, age-adjusted tumor incidence, and survival rates were determined. A total of 7201 cases of malignant carcinoid were identified. Pulmonary and gastrointestinal carcinoid tumors comprised 82% of all carcinoids encountered. The mean age was 64.4 +/- 0.15 years. Stratified by location, there were 3000 (51.4%) foregut carcinoids (including those found in the respiratory tree-2325 in the lung), 2130 (36.5%) midgut carcinoids, and 712 (12.2%) hindgut carcinoids. Second, distinct malignancies were observed in 23% of cases. The total age-adjusted incidence rate has increased from 0.62 per 100,000 in 1980 to 5.17 per 100,000 in 2000. Overall median survival was 21.97 months. The median survival was 19.0 months for foregut carcinoids (excluding those arising in the respiratory tract); 33.9 months for midgut tumors; and 22.7 months for hindgut carcinoids. There was a statistically significant better survival for those with midgut tumors than for those in the other groups (P < 0.001). Age < 60 years, white race, and female sex were all associated with better survival (P < 0.01). The incidence of pulmonary and gastrointestinal carcinoids has dramatically increased since 1981. Tumor location and age > or = 60 years are the strongest predictors of mortality.


The impact of platelet-rich plasma on postoperative outcomes after spinal fusion: a systematic review and meta-analysis.
Platelet-rich plasma (PRP) is a biological agent obtained by centrifuging a sample of blood and retrieving a high concentration of platelets and plasma components. The concentrate is then stimulated for platelet secretion of various growth factors and cytokines. Although it is not widely used in clinical practice, its role in augmenting bony union among patients undergoing spinal fusion has been assessed in several clinical studies. The objective of this study was to perform a systematic review and meta-analysis of the existing literature to determine the efficacy of PRP use in spinal fusion procedures. A comprehensive literature search was conducted using PubMed, Scopus, and EMBASE for studies from all available dates. From eligible studies, data regarding the fusion rate and method of assessing fusion, estimated blood loss (EBL), and baseline and final visual analog scale (VAS) scores were collected as the primary outcomes of interest. Patients were grouped by those undergoing spinal fusion with PRP and bone graft (PRP group) and those only with bone graft (graft-only group). The literature search resulted in 207 articles. Forty-five full-text articles were screened, of which 11 studies were included, resulting in a meta-analysis including 741 patients. Patients without PRP were more likely to have a successful fusion at the last follow-up compared with those with PRP in their bone grafts (OR 0.53, 95% CI 0.34-0.84; p = 0.006). There was no statistically significant difference with regard to change in VAS scores (OR 0.00, 95% CI -2.84 to 2.84; p > 0.99) or change in EBL (OR 3.67, 95% CI -67.13-74.48; p = 0.92) between the groups. This study found that the additional use of PRP was not associated with any significant improvement in patient-reported outcomes and was actually found to be associated with lower fusion rates compared with standard grafting techniques. Thus, PRP may have a limited role in augmenting spinal fusion.


Gastrointestinal Hemorrhage Secondary to Duodenal Cystic Dystrophy in Heterotopic Pancreas.
Cystic dystrophy of the duodenal wall (CDDW) is a complication of heterotopic pancreatic tissue located in the wall of the gastrointestinal tract, characterized by the presence of multiple small cysts, usually found in the wall of the second part of the duodenum. Gastrointestinal hemorrhage due to CDDW is a rare complication. We report the case of a 50-year-old man who was admitted to our hospital for persistent vomiting. The imaging tests confirmed the diagnosis of CDDW. During his stay in hospital, the patient had a gastrointestinal hemorrhage secondary to this disorder, which made it necessary to perform a Roux-en-Y gastrojejunostomy (Billroth III).


Trends in Advance Care Planning in Patients With Cancer: Results From a National Longitudinal Survey.
Advance care planning (ACP) may prevent end-of-life (EOL) care that is nonbeneficial and discordant with patient wishes. Despite long-standing recognition of the merits of ACP in oncology, it is unclear whether participation in ACP by patients with cancer has increased over time. To characterize trends in durable power of attorney (DPOA) assignment, living will creation, and participation in discussions of EOL care preferences and to explore associations between ACP subtypes and EOL treatment intensity as reflected in EOL care decisions and terminal hospitalizations. We analyzed prospectively collected survey data from 1985 next-of-kin surrogates of Health and Retirement Study (HRS) participants with cancer who died between 2000 and 2012, including data from in-depth "exit" interviews conducted with the surrogates after the participant's death. The HRS is a nationally representative, biennial, longitudinal panel study of US residents older than 50 years. Trends in ACP subtypes were tested, and multivariable logistic regression models examined for associations between ACP subtypes and measures of treatment intensity. Trends in the surrogate-reported frequency of DPOA assignment, living will creation, and participation in discussions of EOL care preferences; associations between ACP subtypes and both surrogate-reported EOL care decisions and terminal hospitalizations. From 2000 to 2012, there was an increase in DPOA assignment (52% to 74%, P = .03), without significant change in use of living wills (49% to 40%, P = .63) or EOL discussions (68% to 60%, P = .62). Surrogate reports that patients received "all care possible" at EOL increased during the period (7% to 58%, P = .004), and rates of terminal hospitalizations were unchanged (29% to 27%, P = .70). Limiting or withholding treatment was associated with living wills (adjusted odds ratio [AOR], 2.51; 95% CI, 1.53-4.11; P < .001) and EOL discussions (AOR, 1.93; 95% CI, 1.53-3.14; P = .002) but not with DPOA assignment. Use of DPOA increased significantly between 2000 and 2012 but was not associated with EOL care decisions. Importantly, there was no growth in key ACP domains such as discussions of care preferences. Efforts that bolster communication of EOL care preferences and also incorporate surrogate decision makers are critically needed to ensure receipt of goal-concordant care.


Regulation of meiotic entry and gonadal sex differentiation in the human: normal and disrupted signaling.
Meiosis is a unique type of cell division that is performed only by germ cells to form haploid gametes. The switch from mitosis to meiosis exhibits a distinct sex-specific difference in timing, with female germ cells entering meiosis during fetal development and male germ cells at puberty when spermatogenesis is initiated. During early fetal development, bipotential primordial germ cells migrate to the forming gonad where they remain sexually indifferent until the sex-specific differentiation of germ cells is initiated by cues from the somatic cells. This irreversible step in gonadal sex differentiation involves the initiation of meiosis in fetal ovaries and prevention of meiosis in the germ cells of fetal testes. During the last decade, major advances in the understanding of meiosis regulation have been accomplished, with the discovery of retinoic acid as an inducer of meiosis being the most prominent finding. Knowledge about the molecular mechanisms regulating meiosis signaling has mainly been established by studies in rodents, while this has not yet been extensively investigated in humans. In this review, the current knowledge about the regulation of meiosis signaling is summarized and placed in the context of fetal gonad development and germ cell differentiation, with emphasis on results obtained in humans. Furthermore, the consequences of dysregulated meiosis signaling in humans are briefly discussed in the context of selected pathologies, including testicular germ cell cancer and some forms of male infertility.


Comparison of repositioning maneuvers for benign paroxysmal positional vertigo of posterior semicircular canal: advantages of hybrid maneuver.
The prevalence of benign paroxysmal positional vertigo (BPPV) is becoming more frequent in elderly population. The presence of comorbid factors has to be considered before assessment as well as before commencing any repositioning treatment. Our aims were evaluation of the maneuvers efficacy and evaluation of the applicability of hybrid maneuver (HM) in patients with physical limitation. This is a randomized study in 2 tertiary referral centers. This is a therapeutic intervention. All consecutive patients with diagnosis of BPPV of posterior canal matching the inclusion criteria were enrolled. Patients underwent treatment soon after the initial diagnosis in all cases with a repositioning maneuver. The maneuver was casually selected among Semont, Epley, and hybrid. Patients were divided into 3 groups according to the maneuver adopted. Eighty-eight patients with posterior canal BPPV were enrolled for treatment. Fisher exact test showed that no statistical differences exist between HM and other maneuvers in terms of efficacy. Latency of repositioning nystagmus appeared longer in HM in comparison with other maneuvers (P < .05). Efficacy of maneuvers used for BPPV decreases in case of cupulolithiasis (P < .0001). We found no relationship between age, sex, and length of disturbance on response to maneuvers. All maneuvers evaluated demonstrated similar efficacy. The HM, as our data showed, allows us to obtain a good percentage of success similar to most maneuvers used. It is also more comfortable for the patients with hip or neck functional limitation allowing an effective treatment of the posterior canal BPPV.


Towards understanding the tandem mass spectra of protonated oligopeptides. 1: mechanism of amide bond cleavage.
The mechanism of the cleavage of protonated amide bonds of oligopeptides is discussed in detail exploring the major energetic, kinetic, and entropy factors that determine the accessibility of the b(x)-y(z) (Paizs, B.; Suhai, S. Rapid Commun. Mass Spectrom. 2002, 16, 375) and "diketopiperazine" (Cordero, M. M.; Houser, J. J.; Wesdemiotis, C. Anal. Chem. 1993, 65, 1594) pathways. General considerations indicate that under low-energy collision conditions the majority of the sequence ions of protonated oligopeptides are formed on the b(x)-y(z) pathways which are energetically, kinetically, and entropically accessible. This is due to the facts that (1).the corresponding reactive configurations (amide N protonated species) can easily be formed during ion excitation, (2). most of the protonated nitrogens are stabilized by nearby amide oxygens making the spatial arrangement of the two amide bonds (the protonated and its N-terminal neighbor) involved in oxazolone formation entropically favored. On the other hand, formation of y ions on the diketopiperazine pathways is either kinetically or energetically or entropically controlled. The energetic control is due to the significant ring strain of small cyclic peptides that are co-formed with y ions (truncated protonated peptides) similar in size to the original peptide. The entropy control precludes formation of y ions much smaller than the original peptide since the attacking N-terminal amino group can rarely get close to the protonated amide bond buried by amide oxygens. Modeling the b(x)-y(z) pathways of protonated pentaalanine leads for the first time to semi-quantitative understanding of the tandem mass spectra of a protonated oligopeptide. Both the amide nitrogen protonated structures (reactive configurations for the amide bond cleavage) and the corresponding b(x)-y(z) transition structures are energetically more favored if protonation occurs closer to the C-terminus, e.g., considering these points the Ala(4)-Ala(5) amide bond is more favored than Ala(3)-Ala(4), and Ala(3)-Ala(4) is more favored than Ala(2)-Ala(3). This fact explains the increasing ion abundances observed for the b(2)/y(3), b(3)/y(2), and b(4)/y(1) ion pairs in the metastable ion and low-energy collision induced mass spectra (Yalcin, T.; Csizmadia, I. G.; Peterson, M. B.; Harrison, A. G. J. Am. Soc. Mass Spectrom. 1996, 7, 233) of protonated pentaalanine. A linear free-energy relationship is used to approximate the ratio of the b(x) and y(z) ions on the particular b(x)-y(z) pathways. Applying the necessary proton affinities such considerations satisfactorily explain for example dominance of the b(4) ion over y(1) and the similar b(3) and y(2) ion intensities observed for the metastable ion and low-energy collision induced mass spectra.


The orphan receptor Rev-ErbA alpha activates transcription via a novel response element.
Rev-ErbA alpha (Rev-Erb) is a nuclear hormone receptor-related protein encoded on the opposite strand of the alpha-thyroid hormone receptor (TR) gene. This unusual genomic arrangement may have a regulatory role, but the conservation of human and rodent Rev-Erb amino acid sequences suggests that the protein itself has an important function, potentially as a sequence-specific transcriptional regulator. However, despite its relationship to the TR, Rev-Erb bound poorly to TR binding sites. To determine its DNA-binding specificity in an unbiased manner, Rev-Erb was synthesized in Escherichia coli, purified, and used to select specific binding-sites from libraries of random double-stranded DNA sequences. We found that Rev-Erb binds to a unique site consisting of a specific 5-bp A/T-rich sequence adjacent to a TR half-site. Rev-Erb contacts this entire asymmetric 11-bp sequence, which is the longest nonrepetitive element specifically recognized by a member of the thyroid/steroid hormone receptor superfamily, and mutations in either the A/T-rich or TR half-site regions abolished specific binding. The binding specificity of wild-type Rev-Erb was nearly identical to that of C- and N-terminally truncated forms. This binding was not enhanced by retinoid X receptor, TR, or other nuclear proteins, none of which formed heterodimers with Rev-Erb. Rev-Erb also appeared to bind to the selected site as a monomer. Furthermore, Rev-Erb activates transcription through this binding site even in the absence of exogenous ligand. Thus, Rev-Erb is a transcriptional activator whose properties differ dramatically from those of classical nuclear hormone receptors, including the TR encoded on the opposite strand of the same genomic locus.


Haemostatic dressings in prehospital care.
Massive haemorrhage still accounts for up to 40% of mortality after traumatic injury. The importance of limiting blood loss after injury in order to prevent its associated complications has led to rapid advances in the development of dressings for haemostatic control. Driven by recent military conflicts, there is increasing evidence to support their role in the civilian prehospital care environment. This review aims to summarise the key characteristics of the haemostatic dressings currently available on the market and provide an educational review of the published literature that supports their use. Medline and Embase were searched from start to January 2012. Other sources included both manufacturer and military publications. Agents not designed for use in prehospital care or that have been removed from the market due to significant safety concerns were excluded. The dressings reviewed have differing mechanisms of action. Mineral based dressings are potent activators of the intrinsic clotting cascade resulting in clot formation. Chitosan based dressings achieve haemostasis by adhering to damaged tissues and creating a physical barrier to further bleeding. Acetylated glucosamine dressings work via a combination of platelet and clotting cascade activation, agglutination of red blood cells and local vasoconstriction. Anecdotal reports strongly support the use of haemostatic dressings when bleeding cannot be controlled using pressure dressings alone; however, current research focuses on studies conducted using animal models. There is a paucity of published clinical literature that provides an evidence base for the use of one type of haemostatic dressing over another in humans.


Duplicated genes evolve slower than singletons despite the initial rate increase.
Gene duplication is an important mechanism that can lead to the emergence of new functions during evolution. The impact of duplication on the mode of gene evolution has been the subject of several theoretical and empirical comparative-genomic studies. It has been shown that, shortly after the duplication, genes seem to experience a considerable relaxation of purifying selection. Here we demonstrate two opposite effects of gene duplication on evolutionary rates. Sequence comparisons between paralogs show that, in accord with previous observations, a substantial acceleration in the evolution of paralogs occurs after duplication, presumably due to relaxation of purifying selection. The effect of gene duplication on evolutionary rate was also assessed by sequence comparison between orthologs that have paralogs (duplicates) and those that do not (singletons). It is shown that, in eukaryotes, duplicates, on average, evolve significantly slower than singletons. Eukaryotic ortholog evolutionary rates for duplicates are also negatively correlated with the number of paralogs per gene and the strength of selection between paralogs. A tally of annotated gene functions shows that duplicates tend to be enriched for proteins with known functions, particularly those involved in signaling and related cellular processes; by contrast, singletons include an over-abundance of poorly characterized proteins. These results suggest that whether or not a gene duplicate is retained by selection depends critically on the pre-existing functional utility of the protein encoded by the ancestral singleton. Duplicates of genes of a higher biological import, which are subject to strong functional constraints on the sequence, are retained relatively more often. Thus, the evolutionary trajectory of duplicated genes appears to be determined by two opposing trends, namely, the post-duplication rate acceleration and the generally slow evolutionary rate owing to the high level of functional constraints.


Predicting Progression to Advanced Age-Related Macular Degeneration from Clinical, Genetic, and Lifestyle Factors Using Machine Learning.
Current prediction models for advanced age-related macular degeneration (AMD) are based on a restrictive set of risk factors. The objective of this study was to develop a comprehensive prediction model applying a machine learning algorithm allowing selection of the most predictive risk factors automatically. Two population-based cohort studies. The Rotterdam Study I (RS-I; training set) included 3838 participants 55 years of age or older, with a median follow-up period of 10.8 years, and 108 incident cases of advanced AMD. The Antioxydants, Lipids Essentiels, Nutrition et Maladies Oculaires (ALIENOR) study (test set) included 362 participants 73 years of age or older, with a median follow-up period of 6.5 years, and 33 incident cases of advanced AMD. The prediction model used the bootstrap least absolute shrinkage and selection operator (LASSO) method for survival analysis to select the best predictors of incident advanced AMD in the training set. Predictive performance of the model was assessed using the area under the receiver operating characteristic curve (AUC). Incident advanced AMD (atrophic, neovascular, or both), based on standardized interpretation of retinal photographs. The prediction model retained (1) age, (2) a combination of phenotypic predictors (based on the presence of intermediate drusen, hyperpigmentation in one or both eyes, and Age-Related Eye Disease Study simplified score), (3) a summary genetic risk score based on 49 single nucleotide polymorphisms, (4) smoking, (5) diet quality, (6) education, and (7) pulse pressure. The cross-validated AUC estimation in RS-I was 0.92 (95% confidence interval [CI], 0.88-0.97) at 5 years, 0.92 (95% CI, 0.90-0.95) at 10 years, and 0.91 (95% CI, 0.88-0.94) at 15 years. In ALIENOR, the AUC reached 0.92 at 5 years (95% CI, 0.87-0.98). In terms of calibration, the model tended to underestimate the cumulative incidence of advanced AMD for the high-risk groups, especially in ALIENOR. This prediction model reached high discrimination abilities, paving the way toward making precision medicine for AMD patients a reality in the near future.


Abnormalities of carbohydrate and lipid metabolism in Dahl rats.
Plasma glucose, insulin, and triglyceride concentration, blood pressure, and insulin action on isolated adipocytes were determined in weight-matched Sprague-Dawley, Dahl salt-resistant, and Dahl salt-sensitive rats. Blood pressure and plasma glucose concentrations were not significantly different in the three groups. However, Dahl salt-sensitive rats had significantly higher plasma insulin (39 +/- 2 microunits/ml) and triglyceride (213 +/- 11 mg/dl) concentrations than did Sprague-Dawley rats (27 +/- 2 microunits/ml and 101 +/- 6 mg/dl, respectively). Values for insulin (34 +/- 4 microunits/ml) and triglyceride (159 +/- 11 mg/dl) were intermediate in Dahl salt-resistant rats. In contrast, maximal insulin-stimulated glucose transport was significantly lower in adipocytes isolated from Dahl salt-sensitive as compared with Sprague-Dawley rats (400 +/- 16 versus 523 +/- 14 fl/cell/sec), with Dahl salt-resistant rats again having intermediate values. However, the ability of insulin to maximally inhibit catecholamine-stimulated lipolysis was similar in all three groups, averaging approximately 20% of the activity present in the absence of insulin. All of these differences were seen when the rats were eating conventional chow and did not change in Dahl rats after 2 weeks of an 8% NaCl diet. On the other hand, the predicted rise in blood pressure took place in Dahl salt-sensitive rats, increasing from 147 +/- 4 to 181 +/- 6 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS)


SU-E-T-239: In Vivo Dosimetry with Surface Diodes during Total Body Irradiation: A Patient Thickness Factor to Correct Midline Dose.
In vivo dosimetry (IVD) assessment of treatment dose is important when delivering total body irradiation (TBI). One method is to average AP and PA surface diode measurements and compare them to prescribed midline doses. We designed phantom studies to examine the impact of patient thickness on surface IVD measurements under TBI conditions. Phantom studies were designed to assess the effects of patient thickness on diode IVD. Sun Nuclear QED diodes with inherent buildup were placed on anterior and posterior surfaces of a solid water phantom. Phantom thickness was varied between 20 and 40 cm. A PTW farmer chamber was inserted in the center of the phantom at 425 SSD to reflect prescribed midline dose, and 50 cGy was delivered to midline with 18 MV photons. Averaged entrance and exit diode doses were then compared to farmer chamber measurements of phantom midline dose. A trend of increased deviation with increasing umbilicus thickness was observed between averaged surface diodes and midline farmer chamber measurements. Averaged surface diode dose ranged from 49.6 cGy (20 cm thickness) to 52.1 cGy (40 cm thickness). Interpolation of diode measurements to midline resulted in linear overestimation of delivered dose relative to farmer chamber measurements at midline, up to 6.8% at 40 cm umbilicus thickness. Accurate in vivo dosimetry at time of patient TBI is important to allow individual correction of MU exposure and tissue compensation. Without patient thickness correction, overresponse of surface diodes may lead to unnecessary clinical intervention to treatment MU or compensation and insufficient midline dose. Additionally, SAD setup is preferable to SSD setup to minimize thickness non-linearity. In conclusion, thickness correction factors should be used to generate expected diode readings for patients with thickness greater than 30 cm.


A third class of avian sarcoma viruses, defined by related transformation-specific proteins of Yamaguchi 73 and Esh sarcoma viruses.
The gag-linked transformation-specific protein (polyprotein) p80 of Esh avian sarcoma virus (ESV) has been compared by tryptic peptide mapping with the homologous protein p90 of Yamaguchi 73 avian sarcoma virus (Y73). p80 of ESV and p90 of Y73 were found to share all four of their major nonstructural, transformation-specific, methionine-containing peptides and to have at least seven cysteine-containing transformation-specific peptides in common. Two nonstructural cysteine-containing peptides unique for ESV p80 and three specific for Y73 p90 were also identified. None of these peptides were found in the transforming gene product pp60src of Rous sarcoma virus (RSV) or in the transformation-specific polyproteins p105 of avian sarcoma virus PRCII (PRCII) or p140 of Fujinami sarcoma virus (FSV). ESV p80 and Y73 p90 are phosphorylated, and their tryptic phosphopeptides appear to be identical. In each polyprotein two major phosphopeptides were demonstrated, one containing phosphoserine, the other phosphotyrosine. The latter serves as phosphoacceptor for the protein kinase activities (ATP:protein phosphotransferase, EC 188.8.131.52) associated with p80 and p90. These protein kinase activities were found to be functionally indistinguishable but could be easily distinguished from the activities associated with PRCII p105 and FSV p140 on the basis of their cation requirement and target site specificity. On that basis also, p80/p90-associated protein kinases were found to be more similar to the enzymatic activity of pp60src than to those associated with the PRCII and FSV transformation-specific polyproteins. These results document a close genetic relationship between the two independently isolated sarcoma viruses Y73 and ESV. On the basis of the relatedness of transformation-specific proteins, ESV and Y73 constitute class III of avian sarcoma viruses, with class I containing the various strains of RSV and class II encompassing FSV and PRCII.


Resistive simulated weightbearing exercise with whole body vibration reduces lumbar spine deconditioning in bed-rest.
Randomized controlled trial. Determine the effectiveness a resistive exercise countermeasure with whole-body vibration in relation to lumbo-pelvic muscle and spinal morphology changes during simulated spaceflight (bed-rest). Spinal lengthening, flattening of the spinal curves, increases in disc size, and muscle atrophy are commonly seen in spaceflight simulation. This may represent a risk for low back injury. Consideration of exercise countermeasures against these changes is critical for success of long-term spaceflight missions. Twenty healthy male subjects underwent 8-weeks of bed-rest with 6-months follow-up and were randomly allocated to an inactive control or countermeasure exercise group. Magnetic resonance imaging of the lumbo-pelvic region was conducted at regular time-points during and after bed-rest. Using uniplanar images at L4, cross-sectional areas of the multifidus, lumbar erector spinae, quadratus lumborum, psoas, anterolateral abdominal, and rectus abdominis muscles were measured. Sagittal scans were used to assess lumbar spine morphology (length, sagittal disc area and height, and intervertebral angles). The countermeasure group exhibited less multifidus muscle atrophy (P = 0.024) and its atrophy did not persist long-term as in the control group (up to 3-months; P < 0.006). Spinal lengthening (P = 0.03) and increases in disc area (P = 0.041) were also reduced. Significant partial correlations (P < 0.001) existed between spinal morphology and muscle cross-sectional area changes. The resistive vibration exercise countermeasure reduced, but did not entirely prevent, multifidus muscle atrophy and passive spinal tissue deconditioning during bed-rest. Atrophy of the multifidus muscles was persistent long-term in the inactive subjects. Future work could consider closer attention to spinal posture during exercise and optimizing exercise dose.


Influence of epidermal growth factor on bovine pancreatic duct cell bicarbonate.
Epidermal growth factor (EGF) is secreted in pancreatic juice and its receptor is expressed on pancreatic duct cells (PDCs), suggesting a physiological role which has yet to be defined. Here we examined the effects of EGF on bicarbonate production and carbonic anhydrase (CA) activity in a PDC explant model. Bovine main PDCs were prepared and maintained in culture as explants. Levels of CA expression, phosphorylation, and enzymatic activity were measured in resting cells and compared to that of cells exposed to 10 nM secretin, 10 nM EGF, or both. Bicarbonate production was measured using the autoburette pH titration technique. CA protein levels were unchanged with any treatment, but enzyme activity increased by 180% with secretin treatment and was reduced by 54% with EGF. The combination treatment led to a synergistic increase 240% above basal. EGF alone did not affect bicarbonate secretion, but the normal increase observed with secretin stimulation (1.3 +/- 0.4 to 2.9 +/- 0.6 micromol/h/cm(2)) was abolished by acute EGF pretreatment. On the other hand, EGF pretreatment for 24 h significantly increased basal and stimulated secretion (2.2 +/- 0.5 and 3.8 +/- 0.5, respectively) compared to controls. EGF exerts a regulatory role on bicarbonate secretion by the pancreatic duct epithelium, independent of its effect on CA activity. Its inhibition of stimulated bicarbonate secretion could play a protective role in the setting of pancreatic inflammation, where increased levels of EGF are associated with reduced pancreatic juice production.


Familial abnormality of erythrocyte cation transport in essential hypertension.
Cation transport across the red-cell membrane was studied in subjects with essential hypertension and their relatives using rubidium-86 as an analogue of potassium. The activity of the ouabain-sensitive sodium-potassium pump was significantly greater in patients with untreated essential hypertension than in controls (p less than 0.001). No clear separation was seen between normotensive and hypertensive subjects. Activity of the sodium-potassium pump was also increased in a proportion of normotensive relatives of subjects with essential hypertension. Rubidium uptake was significantly lower in normotensive black subjects than in normotensive whites, the difference being in a ouabain-resistant pathway of cation transport. These results provide further evidence that a defect in membrane cation transport contributes to the pathogenesis of essential hypertension.


Enhanced antitumor efficacy by the combination of emodin and gemcitabine against human pancreatic cancer cells via downregulation of the expression of XIAP in vitro and in vivo.
XIAP and NF-κB play an important role in chemotherapy resistance in pancreatic cancer. The purpose of this study was to explore the role of XIAP and NF-κB in potentiating the antitumor effect of gemcitabine by emodin in pancreatic cancer. SW1990 cells were treated by sodium chloride, gemcitabine, emodin or their combination (gemcitabine plus emodin). Cellular proliferation and apoptosis were detected by Cell Counting kit-8 (CCK-8) assay and flow cytometry in vitro. The combination therapy more significantly inhibited SW1990 cell growth and induced a higher percentage of apoptosis than monotherapy. Gemcitabine upregulated the expression of XIAP and NF-κB, while emodin or emodin plus gemcitabine downregulated them compared to the control group in vitro. SW1990 cells were used to establish orthotopic pancreatic tumor models in nude mice. Tumor-bearing mice were treated with sodium chloride, emodin, gemcitabine or their combination. After being treated for 4 weeks, the nude mice were imaged with high-resolution positron emission tomography (microPET) and fluorine-18-labeled fluorodeoxyglucose (18F-FDG) to detect the tumor/non-tumor ratio (T/NT ratio) and standard uptake value (SUV). The mice were sacrificed to determine tumor weight. The combination of emodin and gemcitabine showed more significant reduction in the T/NT ratio, SUV and tumor weight compared to monotherapy. The mRNA levels and the protein expression of XIAP and NF-κB were upregulated in the gemcitabine group, while they were downregulated in the emodin group and the combination group in vivo. Ki-67 prolif-eration index and TUNEL assay results also showed that emodin enhanced tumor apoptosis induced by gemcitabine in vivo. This study suggests that emodin enhances the antitumor effect of gemcitabine in SW1990 pancreatic cancer in vitro and in vivo, which may be via the downregulation of NF-κB expression, thus inhibiting the expression of XIAP.


[Finite element analysis of male lower urinary tract based on the collodion slice images].
Males typically have high rates of morbidity of primary bladder neck obstruction, while the existing urodynamic examination is invasive and more likely to cause false diagnosis. To build a non-invasive biomechanical detecting system for the male lower urinary tract, a finite element model for male lower urinary tract based on the collodion slice images of normal male lower urinary tract was constructed, and the fluid-structure interaction of the lower urinary tract was simulated based on the real urination environment. The finite element model of the lower urinary tract was validated by comparing the clinical experiment data with the simulation result. The stress, flow rate and deformation of the lower urinary tract were analyzed, and the results showed that the Von Mises stress and the wall shear stress at the membrane sphincter in the normal male lower urinary tract model reached a peak, and there was nearly 1 s delay than in the bladder pressure, which helped to validate the model. This paper lays a foundation for further research on the urodynamic response mechanism of the bladder pressure and flow rate of the lower urinary tract obstruction model, which can provide a theoretical basis for the research of non-invasive biomechanical detecting system.


Fluid-Structure Interaction Study of Transcatheter Aortic Valve Dynamics Using Smoothed Particle Hydrodynamics.
Computational modeling of heart valve dynamics incorporating both fluid dynamics and valve structural responses has been challenging. In this study, we developed a novel fully-coupled fluid-structure interaction (FSI) model using smoothed particle hydrodynamics (SPH). A previously developed nonlinear finite element (FE) model of transcatheter aortic valves (TAV) was utilized to couple with SPH to simulate valve leaflet dynamics throughout the entire cardiac cycle. Comparative simulations were performed to investigate the impact of using FE-only models vs. FSI models, as well as an isotropic vs. an anisotropic leaflet material model in TAV simulations. From the results, substantial differences in leaflet kinematics between FE-only and FSI models were observed, and the FSI model could capture the realistic leaflet dynamic deformation due to its more accurate spatial and temporal loading conditions imposed on the leaflets. The stress and the strain distributions were similar between the FE and FSI simulations. However, the peak stresses were different due to the water hammer effect induced by the fluid inertia in the FSI model during the closing phase, which led to 13-28% lower peak stresses in the FE-only model compared to that of the FSI model. The simulation results also indicated that tissue anisotropy had a minor impact on hemodynamics of the valve. However, a lower tissue stiffness in the radial direction of the leaflets could reduce the leaflet peak stress caused by the water hammer effect. It is hoped that the developed FSI models can serve as an effective tool to better assess valve dynamics and optimize next generation TAV designs.


Attractiveness of blonde women in evolutionary perspective: studies with two Polish samples.
An experimental study was undertaken to assess the phenomenon of male preference for blondes. In the first study, 360 Polish men ages 18 to 46 years were asked to assess the attractiveness of the presented stimuli using a 9-point scale. Stimuli were 9 different pictures of the same women whose ages (about 20, 30, and 40 years old) and hair colors (blonde, brown, and brunette) were manipulated. Pictures of blonde-haired women were generally rated as younger than the others. The attractiveness ratings of female faces changed with age and hair color. Still, only the 30-yr.-old woman with blonde hair was rated as significantly more attractive than those with brown or brunette hair. In a second study (the analysis of 500 Internet advertisements) mature women dyed their hair blonde more frequently. These results are analyzed with regard to the evolutionarily formed male preference for younger females.


Thermal reactivity of neutral and oxidized ferrocenyl-substituted enediynes.
The coupling of two equivalents of ethynylferrocene (2) with one equivalent of 1,2-diiodocyclohexene (1) and 1,2-diiodobenzene (4) using Sonogashira cross-coupling conditions led to 1,2-bis(ferrocenylethynyl)cyclohexene (3) and 1,2-bis(ferrocenylethy-nyl)benzene (5), respectively. At high temperatures enediynes 3 and 5 showed exothermic signals in differential scanning calorimetry (DSC) measurements, suggestive of intramolecular diradicaloid ring formation (Bergman (C1-C6) or Schreiner-Pascal (C1-C5) cyclizations). The oxidation of 3 and 5 to the mono-oxidized enediynes 3+ and 5+ decreased the onset temperatures drastically. Equally, 1-ferrocenylethynyl-2-(p-nitro-phenyl)ethynylbenzene (8) displayed a significant decrease in the onset temperature after oxidation to 8+. Because the insoluble nature of the polymeric material formed in the thermolysis of the oxidized enediynes prevented characterization, the origin of this drastic effect was studied by DFT. Contrary to expectations, one-electron oxidation does not lower the barrier for intramolecular cyclization. Rather, the computations suggest that the polymerization is initiated by a bimolecular process.


Double-blind controlled trial of lecithinized superoxide dismutase in patients with idiopathic interstitial pneumonia - short term evaluation of safety and tolerability.
Idiopathic interstitial pneumonias such as idiopathic pulmonary fibrosis or fibrotic nonspecific interstitial pneumonia are irreversible progressive pulmonary diseases that often have fatal outcomes. Although the etiology of idiopathic interstitial pneumonias is not yet fully understood, anti-fibrotic and anti-inflammatory agents have shown limited therapeutic effectiveness. Reactive oxygen species and their cytotoxic effects on the lung epithelial cells have been reported to participate in the pathophysiology of the disease. Because superoxide dismutase catalyzes the detoxification of reactive oxygen species, we developed lecithinized superoxide dismutase for the treatment of patients with idiopathic interstitial pneumonias. A multicenter, randomized, placebo-controlled trial was conducted as a pilot study to investigate the safety and effectiveness of 40 or 80 mg lecithinized superoxide dismutase in patients with progressive idiopathic interstitial pneumonias who presented with either idiopathic pulmonary fibrosis or corticosteroid-resistant fibrotic nonspecific interstitial pneumonia and showed arterial oxygen tension compatible with stage III or IV on the Japanese severity grading scale for idiopathic interstitial pneumonias. Before and following infusion of lecithinized superoxide dismutase for 28 days, the primary endpoint of forced vital capacity and the secondary endpoints of lactate dehydrogenase, surfactant protein-A, surfactant protein-D and Krebs von den Lungen-6 levels were measured in the serum. The primary endpoint of forced vital capacity did not improve significantly in the lecithinized superoxide dismutase groups in comparison with the placebo group. The secondary endpoints of lactate dehydrogenase and surfactant protein-A levels were significantly attenuated by 28 days in the higher-dose (80 mg) group. However, these changes returned to the baseline levels by 56 days after the cessation of lecithinized superoxide dismutase. Adverse events and mortality in the drug-treated groups did not differ from those in the placebo group. Treatment with lecithinized superoxide dismutase is safe and improves the levels of serum markers such as lactate dehydrogenase and surfactant protein-A in patients with advanced idiopathic interstitial pneumonias with severe respiratory dysfunction. Considering the results of the current study, further investigations into the effects and treatment potential of long-term administration of lecithinized superoxide dismutase may be warranted. University hospital Medical Information Network (UMIN) clinical trials registry no. 000000752.


Quantum-dot-decorated robust transductable bioluminescent nanocapsules.
Bioluminescence, due to its high sensitivity, has been exploited in various analytical and imaging applications. In this work, we report a highly stable, cell-transductable, and wavelength-tunable bioluminescence system achieved with an elegant and simple design. Using aqueous in situ polymerization on a bioluminescent enzyme anchored with polymerizable vinyl groups, we obtained nanosized core-shell nanocapsules with the enzyme as the core and a cross-linked thin polymer net as the shell. These nanocapsules possess greatly enhanced stability, retained bioactivity, and a readily engineered surface. In particular, by incorporating polymerizable amines in the polymerization, we endowed the nanocapsules with efficient cell-transduction and sufficient conjugation sites for follow-up modification. Following in situ polymerization, decorating the polymer shell with fluorescent quantum dots allowed us to access a continuous tunable wavelength, which extends the application of such bioluminescent nanocapsules, especially in deep tissue. In addition, the unique core-shell structure and adequate conjugation sites on surface enabled us to maximize the BRET efficiency by adjusting the QD/enzyme conjugation ratio.


Overexpression of fucosyltransferase IV promotes A431 cell proliferation through activating MAPK and PI3K/Akt signaling pathways.
Lewis Y (LeY) is a carbohydrate tumor-asssociated antigen. The majority of cancer cells derived from epithelial tissue express LeY type difucosylated oligosaccharide. Fucosyltransferase IV (FUT4) is an essential enzyme that catalyzes the synthesis of LeY oligosaccharide. Our previous studies have shown that FUT4 overexpression promotes A431 cell proliferation, but the mechanism is still largely unknown. Herein, we investigated the role of the mitogen-activated protein kinases (MAPKs) and phosphoinositide-3 kinase (PI3K)/Akt signaling pathways on FUT4-induced cell proliferation. Results show that overexpression of FUT4 increases the phosphorylation of ERK1/2, p38 MAPK, and PI3K/Akt. Inhibitors of PI3K (LY294002 and Wortmannin) prevented the phosphorylation of ERK1/2, p38 MAPK, and Akt PI3K). Moreover, phosphorylation of Akt is abolished by inhibitors of ERK1/2 (PD98059) and p38 MAPK (SB203580). These data suggested that FUT4 not only activates MAPK and PI3K/Akt signals, but also promotes the crosstalk among these signaling pathways. In addition, FUT4-induced stimulation of cell proliferation correlates with increased cell cycle progression by promoting cells into S-phase. The mechanism involves in increased expression of cyclin D1, cyclin E, CDK 2, CDK 4, and pRb, and decreased level of cyclin-dependent kinases inhibitors p21 and p27, which are blocked by the inhibitors of upstream signal molecules, MAPK and PI3K/Akt. In conclusion, these studies suggest that FUT4 regulates A431 cell growth through controlling cell cycle progression via MAPK and PI3K/Akt signaling pathways.


Environmental correlates of physical activity and dietary behaviours among young people: a systematic review of reviews.
An extensive body of research exists on environmental influences on weight-related behaviours in young people. Existing reviews aimed to synthesize this body of work, but generally focused on specific samples, behaviours or environmental influences and integration of findings is lacking. Hereto, we reviewed 18 reviews representing 671 unique studies, aiming to identify what environmental factors do and do not affect physical activity and dietary behaviours in children and adolescents. Eleven reviews focused exclusively on physical activity, six on diet, and one review focused on both physical activity and dietary behaviours with only small overlap in included studies. Physical activity was more consistently related to school and neighbourhood characteristics than to interpersonal and societal environments. In contrast, interpersonal factors played a pronounced role in dietary behaviours; no school, neighbourhood or societal factors were consistently related to dietary behaviours. This review of reviews adds to the literature by providing a comprehensive synthesis of factors related to physical activity and dietary behaviours that could be targeted in interventions. Moreover, by identifying factors that are unrelated to physical activity and dietary behaviours, this review may help to narrow the scope of future studies and environmental interventions.


Energy expenditure and ventilatory responses during Virasana--a yogic standing posture.
Energy expenditure and ventilatory responses to yogic standing posture of Virasana were studied on 10 healthy men (25-37 years of age). The results of various responses respectively to the horizontal supine, Chair-sitting and Virasana were: Minute Ventilation (VE) 7.64, 8.61 and 18.67 L/min; Respiratory Frequency (FR) 15.71, 15.70 and 21.45 Breath/min; Tidal Volume (VT) 0.496, 0.544 and 0.827 L/min; Oxygen consumption (VO2) 0.127, 0.234 and 0.573 L/min; Carbondioxide Elimination (VCO2) 0.127, 0.134 and 0.420 L/min; Respiratory Exchange Ratio (RER) 0.58, 0.57 and 0.69; Heart Frequency (FH) 65.2, 74.5 and 104.4 beats/min; Oxygen Pulse (O2P) 3.32, 3.17 and 5.45 ml/beat; Ventilatory Equivalent (VE-EQ) 36.78, 37.12 and 33.85; Multiple of Resting VO2 (METS) 0.96, 1.05 and 2.53 and Metabolic Cost (MC) 1.04, 1.13 and 2.76 Cal/min. Virasana posture was characterised by higher VE, FR, VT, VO2, VCO2, FH and O2P with lesser VE-EQ. The observations suggest that Virasana induces temporarily a hypermetabolic state characterised by enhanced sympathetic nervous system activity which gets inhibited during the adoption of resting supine shavasana posture.


Structure-Guided Identification of Black Cohosh (Actaea racemosa) Triterpenoids with In Vitro Activity against Multiple Myeloma.
Black cohosh is a well-established medicinal plant and preparations of its rootstock are used for the treatment of mild climacteric complaints. The compounds considered responsible for the therapeutic effect are triterpene glycosides, characterized by a cycloartane scaffold and a pentose moiety. Because some of these triterpenoids were found to exhibit relevant cytotoxic effects against human breast cancer cells, we decided to investigate their activity on multiple myeloma cell lines NCI-H929, OPM-2, and U266. In a systematic approach, we initially tested three known cytotoxic compounds of three different triterpenoid types, revealing the cimigenol-type triterpenoid as the most active constituent. In a second round, seven naturally occurring cimigenol derivatives were compared with respect to their sugar moiety and their substitution pattern at position C-25, leading to 25-O-methylcimigenol-3-O-α-L-arabinopyranoside as the most potent candidate. Interestingly, not only the methyl group at position C-25 increased the cytotoxic effect but also the arabinose moiety at position C-3 had an impact on the activity. The variety of cimigenol derivatives, moreover, allowed a detailed discussion of their structure-activity relationships, not only for their effect on multiple myeloma cells but also with regard to previous studies on the cytotoxicity of black cohosh triterpenoids.


Retrospective exposure assessment and quality control in an international multi-centre case-control study.
The paper presents the exposure assessment method and quality control procedure used in an international, multi-centre case-control study within a joint Nordic and Italian cohort. This study was conducted to evaluate whether occupational exposure to carcinogens influenced the predictivity of high frequency of chromosomal aberrations (CA) in peripheral lymphocytes for increased cancer risk. Occupational hygienists assessed exposures in each participating country: Denmark, Finland, Italy, Norway and Sweden. The exposure status to a carcinogen or a clastogen was coded in the cohort according to the original CA studies at the time of CA testing, but not for the whole work life. An independent occupational hygienist coordinated harmonization of the assessment criteria and the quality control procedure. The reliability of the exposure assessments was calculated as deviation from the majority of the assessors, as Cohen's kappa and as overall proportion of the agreements. The reassessment of the exposures changed the exposure statuses significantly, when compared with the original cohort. Harmonization of the exposure criteria increased the conformity of the assessments. The prevalence of exposure was higher among the original assessors (the assessor from the same country as the subject) than the average prevalence assessed by the other four in the quality control round. The original assessors classified more job situations as exposed than the others. Several reasons for this are plausible: real country-specific differences, differences in information available to the home assessor and the others and misunderstandings or difficulties in translation of information. To ensure the consistency of exposure assessments in international retrospective case-control studies it is important to have a well-planned study protocol. Due to country-specific environments a hygienist from each participating country is necessary. A quality control study is recommended, to be performed as described, combined with round-table meetings to minimize information bias between the assessors.


A comparison of the concept of patient participation and patients' descriptions as related to healthcare definitions.
PURPOSE. To depict what patients describe as patient participation and whether descriptions of patient participation are affected by gender, age, healthcare contact, and duration of disease. DATA SOURCES. Current patients (n= 362) responded to a questionnaire on participation. DATA SYNTHESIS. Patients' descriptions focused on having knowledge, rather than being informed, and on interacting with health professionals, rather than merely partaking in decision making. CONCLUSIONS. Patients' descriptions of participation correspond with the International Classification of Functioning, Disability and Health's definition, which includes "being involved in a life situation." Healthcare legislation and professionals employ a narrower concept of patient participation as defined by, e.g., Medical Subject Headings. PRACTICE IMPLICATIONS. Findings suggest that health professionals need to embrace what patients describe as participation.


Modeling contribution of shallow groundwater to evapotranspiration and yield of maize in an arid area.
Capillary rise from shallow groundwater can decrease the need for irrigation water. However, simple techniques do not exist to quantify the contribution of capillary flux to crop water use. In this study we develop the Agricultural Water Productivity Model for Shallow Groundwater (AWPM-SG) for calculating capillary fluxes from shallow groundwater using readily available data. The model combines an analytical solution of upward flux from groundwater with the EPIC crop growth model. AWPM-SG was calibrated and validated with 2-year lysimetric experiment with maize. Predicted soil moisture, groundwater depth and leaf area index agreed with the observations. To investigate the response of model, various scenarios were run in which the irrigation amount and groundwater depth were varied. Simulations shows that at groundwater depth of 1 m capillary upward supplied 41% of the evapotranspiration. This reduced to 6% at groundwater depth of 2 m. The yield per unit water consumed (water productivity) was nearly constant for 2.3 kg/m3. The yield per unit water applied (irrigation water productivity) increased with decreasing irrigation water because capillary rise made up in part for the lack of irrigation water. Consequently, using AWPM-SG in irrigation scheduling will be beneficial to save more water in areas with shallow groundwater.


Risk factors for falls with severe fracture in elderly people living in a middle-income country: a case control study.
Fracture after falling has been identified as an important problem in public health. Most studies of risk factors for fractures due to falls have been carried out in developed countries, although the size of the elderly population is increasing fast in middle income countries. The objective of this paper is to identify risk factors for fall related to severe fractures in those aged 60 or more in a middle-income country. A case-control study was carried out in Rio de Janeiro-Brazil based general hospitals between 2002-2003. Two hundred-fifty hospitalised cases of fracture were matched with 250 community controls by sex, age group and living area. Data were collected for socio-demographic variables, health status and drugs used before the fall. A conditional logistic regression model was fitted to identify variables associated with the risk of fall related severe fracture. Low body mass index, cognitive impairment, stroke and lack of urine control were associated with increased risk of severe fall related fractures. Benzodiazepines and muscle relaxants were also related to an increased risk of severe fractures while moderate use of alcohol was associated with reduced risk. Although the association between benzodiazepines and fractures due to fall has been consistently demonstrated for old people, this has not been the case for muscle relaxant drugs. The decision to prescribe muscle relaxants for elderly people should take into account the risk of severe fracture associated with these drugs.


[Occurrence of a hepatitis A outbreak in three neighborhoods of the city of Vitória, Espírito Santo State, and its relation with the quality of the water for human consumption].
This work had as purpose to analyze the possible factors related with the occurrences of Hepatitis in the neighborhoods of Grande Vitória, Estrelinha and Inhanguetá located in the city of Vitória, State of the Espírito Santo, from April the June, 2006, and the quality of the water consumed by the population. The indicators for the water quality used were the Free Residual Chlorine concentrations (FRC) and the presence of thermotolerant coliforms. The microbiological analyses performed in the water consumed for the studied communities did not presented results indicating contamination by thermotolerants coliforms and the percentage of FRC determined between January and June were always above of the minimum limit demanded by the legislation (Act 518/2004). The results showed that the occurrence of a Hepatitis A outbreak can be related with the absence of physical barriers that impede the contact of people with the contaminated water and the practical deficiencies in the hygienic practices adopted, individually, by the local population.


A new cell division operon in Escherichia coli.
At 76 min on the E. coli genetic map there is a cluster of genes affecting essential cellular functions, including the heat shock response and cell division. A combination of in-vivo and in-vitro genetic analysis of cell division mutants suggests that the cell division gene fts E is the second gene in a 3 gene operon. A cold-sensitive mutant, defective in the third gene, is also unable to divide at the restrictive temperature, and we designate this new cell division gene fts X. Another cell division gene, fts S, is very close to, but distinct from, the 3 genes of the operon. The fts E product is a 24.5 Kd polypeptide which shows strong homology with a small group of proteins involved in transport. Both the fts E product and the protein coded by the first gene (fts Y) in the operon have a sequence motif found in a wide range of heterogeneous proteins, including the Ras proteins of yeast. This common domain is indicative of a nucleotide-binding site.


Development and validation of a symptom assessment tool for postmicturition dribble: A prospective, multicenter, observational study in Korea.
Postmicturition dribble (PMD) is a very common symptom in males with lower urinary tract symptoms (LUTS) worldwide, but there is no adequate questionnaire to assess it. Therefore, we developed a questionnaire named the Hallym Post Micturition Dribble Questionnaire (HPMDQ) to assess PMD, and the aim of this study is to validate it. A series of consecutive male patients newly diagnosed with LUTS and over 40 years of age who visited any of 5 medical institutions were included. LUTS were assessed in all patients using the International Prostate Symptom Score (IPSS), and PMD was assessed using the HPMDQ. In total, 2134 male patients aged 40 to 91 years were included in this study. Of these patients, 1088 (51.0%) reported PMD. In the PMD group, the mean values for HPMDQ-Q1, HPMDQ-Q2, HPMDQ-Q3 and HPMDQ total score were 1.39, 1.10, 1.76 and 4.25, respectively. In the non-PMD group, the mean values of these scores were 0, 0.18, 1.52 and 1.58, respectively. The difference in HPMDQ scores between the 2 groups was statistically significant. PMD was significantly associated with the voiding symptoms of LUTS, prostate size and postvoid residual but not with storage symptoms. The HPMDQ, which consists of 5 questions (frequency, severity, bother, quality of life and response to treatment for PMD), was developed, and its use for assessing PMD is validated in this study. It may be a useful tool for further research and in clinical practice for PMD.


Quorum sensing in Vibrio anguillarum: characterization of the vanI/vanR locus and identification of the autoinducer N-(3-oxodecanoyl)-L-homoserine lactone.
Certain gram-negative pathogens are known to control virulence gene expression through cell-cell communication via small diffusible signal molecules termed autoinducers. This intercellular signal transduction mechanism termed quorum sensing depends on the interaction of an N-acylhomoserine lactone (AHL) auto-inducer molecule with a receptor protein belonging to the LuxR family of positive transcriptional activators. Vibrio anguillarum is a gram-negative pathogen capable of causing a terminal hemorrhagic septicemia known as vibriosis in fish such as rainbow trout. In this study, we sought to determine whether V. anguillarum employs AHLs to regulate virulence gene expression. Spent V. anguillarum culture supernatants stimulated bioluminescence in a recombinant lux-based Escherichia coli AHL biosensor strain, whereas they both stimulated and inhibited AHL-mediated violacein pigment production in Chromobacterium violaceum. This finding suggested that V. anguillarum may produce multiple AHL signal molecules. Using high-performance liquid chromatography and high-resolution tandem mass spectrometry, we identified the major V. anguillarum AHL as N-(3-oxodecanoyl)-L-homoserine lactone (ODHL), a structure which was unequivocally confirmed by chemical synthesis. The gene (vanI) responsible for ODHL synthesis was cloned and sequenced and shown to belong to the LuxI family of putative AHL synthases. Further sequencing downstream of vanI revealed a second gene (vanR) related to the LuxR family of transcriptional activators. Although deletion of vanI abolished ODHL synthesis, no reduction of either metalloprotease production or virulence in a fish infection model was observed. However, the vanI mutant remained capable of weakly activating both bioluminescence and violacein in the E. coli and C. violaceum biosensors, respectively, indicating the existence of additional layers of AHL-mediated regulatory complexity.


Treatment of deep venous thrombosis with low-molecular-weight heparins. A meta-analysis.
An intravenous course of unfractionated heparin adjusted on the basis of the activated partial thromboplastin time is the initial treatment of choice for most patients with venous thromboembolism. Recently introduced low-molecular-weight heparin preparations can be administered subcutaneously, once or twice daily, without laboratory monitoring. We quantitatively assessed the relative efficacy and safety of low-molecular-weight heparin vs standard heparin for the initial treatment of deep venous thrombosis. English-language reports of randomized trials were identified through a MEDLINE search (1984 through 1994) and a complementary extensive manual search. Reasons for exclusion from the analysis were no heparin dosage adjustments, the lack of use of objective tests for deep venous thrombosis, duplicate reports, preliminary reports of data later presented in full, dose-ranging studies that used higher doses of low-molecular-weight heparin than are currently in use, and the failure to provide blind end-point assessment. We assessed the incidence of symptomatic recurrent venous thromboembolic disease, the incidence of clinically important bleeding, and mortality. Ten of the 19 identified trials satisfied the predetermined criteria. The relative risk reductions for symptomatic thromboembolic complications (53% [95% confidence interval, 18% to 73%]), clinically important bleeding (68% [95% confidence interval, 31% to 85%]), and mortality (47% [95% confidence interval, 10% to 69%]) were all statistically significantly in favor of low-molecular-weight heparin. Low-molecular-weight heparins administered subcutaneously in fixed doses adjusted for body weight and without laboratory monitoring are more effective and safer than adjusted-dose standard heparin. Since low-molecular-weight heparins may not be interchangeable and the conclusions of our meta-analysis are based mainly on the findings of three trials that used two different low-molecular-weight heparins, definitive randomized controlled trials for the other low-molecular-weight heparins are required.


Loss of heterozygosity on chromosomes 3, 9, 13, and 17, including the retinoblastoma locus, in uveal melanoma.
To identify tumor-suppressor loci that may contribute to the pathogenesis of uveal melanoma. Multiplex fluorescence microsatellite assays were performed on 27 uveal melanomas using markers at 3p25-p26, 3p14.2, 9p21-p23, 13q14, 13q12.3-q13, and 17p13, close to or within the von Hippel Lindau (VHL), fragile histidine triad (FHIT), p16/cyclin-dependent kinase inhibitor 2 (CDKN2A), retinoblastoma (RB1), breast cancer 2 (BRCA2), and p53 tumor suppressor loci, respectively. Further markers on chromosomes 3 and 9 were analyzed individually. Loss of heterozygosity (LOH) was identified in 63% of tumors, most frequently on chromosome 3 (52%), in association with epithelioid cells (P = 0.0002) and microvascular loops (P = 0.0008). In the majority of cases, LOH on chromosome 3 was detected at all informative markers. The second most common alteration was LOH at an RB1 intragenic marker (21% tumors), with retention of a more centromeric 13q marker (near BRCA2). The pattern of LOH on chromosome 9p was consistent with the involvement of a region telomeric to CDKN2A. LOH at TP53 was infrequent. In the majority of cases, chromosome 3 LOH involves an entire chromosome homologue, which hampers identification of the relevant suppressor loci. This LOH correlates with the presence of microvascular loops and epithelioid cells, two of the recognized histologic indicators of poor prognosis. Data for chromosomes 13 and 9 support a role for RB1 in the pathogenesis of uveal melanoma but also raise the possibility of the involvement of additional loci close to RB1 and CDKN2A.


Relation of self-concept in kindergarten and first grade to school adjustment.
The present aim was to investigate changes in self-concept and self-concept stability during children's transition from kindergarten to primary school, and the relationship between children's self-concept and their adjustment to elementary school. 346 children took part in this 2-yr. longitudinal study. The Pictorial Scale of Perceived Competence and Social Acceptance for Young Children was used to assess children's self-concept. Children's school adjustment was assessed using the schools' academic, behavioral, and social-emotional procedures. Children's self-concept appeared relatively stable and the decline in scores appeared to be practically unimportant. Analysis showed that children's school adjustment mainly affects the academic dimension of self-concept.


Isolated and combined risks for anemia in children attending the nurseries of daycare centers.
To identify and quantify isolated and combined risk factors for anemia, providing a comprehensive view of the likelihood of its occurrence. Cross-sectional study with 482 children aged 4 to 29 months attending the nurseries of philanthropic and public daycare centers in the city of São Paulo, Brazil, who participated in two surveys (2004 and 2007). Mothers were interviewed, blood was collected using digital puncture, and anthropometry was performed. Anemia was characterized by hemoglobin levels below 11 g/dL. Unconditional logistic regression was adjusted for anemia risk factors. A value of p < or = 0.05 indicated statistically significant associations. Post-test odds and likelihood ratios were calculated to define post-test probabilities. Epi-Info 2000 and Stata 10.0 software packages were used for statistical analysis. Prevalence of anemia was 43.6% (95%CI 39.1-48.1). The final logistic model included five categorical variables: mother's age less than 28 years (OR = 1.50; p = 0.041), per capita income below half a minimum wage (OR = 1.56; p = 0.029), exclusive breastfeeding less than 2 months (OR = 1.71; p = 0.009), decrease in weight/age z score from birth to survey (OR = 1.47; p = 0.050), and age less than 17 months (OR = 2.44; p < 0,001). Post-test probabilities in the associations of isolated and combined risk factors for anemia ranged from 54.5 to 100%. The probability of anemia progressively increased as the identified risk factors were added. This calculation provides a simple and rapid tool for suspicion of anemia in children both in clinical practice and population screening.


Identification and characterization of repetitive extragenic palindromes (REP)-associated tyrosine transposases: implications for REP evolution and dynamics in bacterial genomes.
Bacterial repetitive extragenic palindromes (REPs) compose a distinct group of genomic repeats. They usually occur in high abundance (>100 copies/genome) and are often arranged in composite repetitive structures - bacterial interspersed mosaic elements (BIMEs). In BIMEs, regularly spaced REPs are present in alternating orientations. BIMEs and REPs have been shown to serve as binding sites for several proteins and suggested to play role in chromosome organization and transcription termination. Their origins are, at present, unknown. In this report, we describe a novel class of putative transposases related to IS200/IS605 transposase family and we demonstrate that they are obligately associated with bacterial REPs. Open reading frames coding for these REP-associated tyrosine transposases (RAYTs) are always flanked by two REPs in inverted orientation and thus constitute a unit reminiscent of typical transposable elements. Besides conserved residues involved in catalysis of DNA cleavage, RAYTs carry characteristic structural motifs that are absent in typical IS200/IS605 transposases. DNA sequences flanking rayt genes are in one third of examined cases arranged in modular BIMEs. RAYTs and their flanking REPs apparently coevolve with each other. The rayt genes themselves are subject to rapid evolution, substantially exceeding the substitution rate of neighboring genes. Strong correlation was found between the presence of a particular rayt in a genome and the abundance of its cognate REPs. In light of our findings, we propose that RAYTs are responsible for establishment of REPs and BIMEs in bacterial genomes, as well as for their exceptional dynamics and species-specifity. Conversely, we suggest that BIMEs are in fact a special type of nonautonomous transposable elements, mobilizable by RAYTs.


Clinical spectrum and molecular characteristics of Klebsiella pneumoniae causing community-acquired extrahepatic abscess.
Klebsiella pneumoniae causes a wide spectrum of infections, including abscess and non-abscess formation. This study investigated the clinical spectrum and molecular characteristics of community-acquired Klebsiella infection with primary extrahepatic abscess. From April 2004 through March 2007, a total of 18 strains of K. pneumoniae, 11 from blood and 7 from focal purulent specimens, were recovered from a medical center in southern Taiwan. The clinical data were collected from medical records. Hypermucoviscosity phenotype was defined as positive string test. The virulence genes, including rmpA (regulator of mucoid phenotype), magA (specific to K1 capsule serotype), k2A (specific to K2 capsule serotype), and kfu (an iron uptake system) were amplified by polymerase chain reaction using specific primers. Twelve men and 6 women with ages ranging from 37 to 74 years were enrolled. Fifteen patients had underlying diabetes mellitus. The duration of hospitalization ranged from 1 to 96 days. Three patients died by the end of treatment. All of the K. pneumoniae strains carried rmpA and 16 strains showed the hypermucoviscosity phenotype. Of the 18 strains, 7 strains were positive for k2A and 4 strains carried magA. kfu was identified in 4 magA-positive strains and 2 magA-negative/k2A-negative strains. Diabetes mellitus was the most frequent underlying disease among our patients. The rmpA and/or hypermucoviscosity phenotype were the most common virulence factors in K. pneumoniae isolates causing extrahepatic abscesses, among which K2 capsule serotype (k2A+) was more prevalent than K1 capsule serotype (magA+).


Crystal structures of the binary and ternary complexes of 7 alpha-hydroxysteroid dehydrogenase from Escherichia coli.
7 alpha-Hydroxysteroid dehydrogenase (7 alpha-HSDH;1 EC 188.8.131.52) is an NAD+-dependent oxidoreductase belonging to the short-chain dehydrogenase/reductase (SDR) 1 family. It catalyzes the dehydrogenation of a hydroxyl group at position 7 of the steroid skeleton of bile acids. The crystal structure of the binary (complexed with NAD+) complex of 7 alpha-HSDH has been solved at 2.3 A resolution by the multiple isomorphous replacement method. The structure of the ternary complex [the enzyme complexed with NADH, 7-oxoglycochenodeoxycholic acid (as a reaction product), and possibly partially glycochenodeoxycholic acid (as a substrate)] has been determined by a difference Fourier method at 1.8 A resolution. The enzyme 7 alpha-HSDH is an alpha/beta doubly wound protein having a Rossmann-fold domain for NAD (H) binding. Upon substrate binding, large conformation changes occur at the substrate binding loop (between the beta F strand and alpha G helix) and the C-terminal segment (residues 250-255). The variable amino acid sequences of the substrate-binding loop appear to be responsible for the wide variety of substrate specificities observed among the enzymes of the SDR family. The crystal structure of the ternary complex of 7 alpha-HSDH, which is the only structure available as the ternary complex among the enzymes of the SDR family, indicates that the highly conserved Tyr159 and Ser146 residues most probably directly interact with the hydroxyl group of the substrates although this observation cannot be definite due to an insufficiently characterized nature of the ternary complex. The strictly conserved Lys163 is hydrogen-bonded to both the 2'- and 3'-hydroxyl groups of the nicotinamide ribose of NAD(H). We propose a new catalytic mechanism possibly common to all the enzymes belonging to the SDR family in which a tyrosine residue (Tyr159) acts as a catalytic base and a serine residue (Ser146) plays a subsidiary role of stabilizing substrate binding.


Reevaluation of autoantibodies to islet cell membrane in IDDM. Failure to detect islet cell surface antibodies using human islet cells as substrate.
Since their demonstration in 1975, ICSAs have been proposed as serological markers and pathogenic elements in IDDM. ICSAs are detected in the sera of most newly diagnosed IDDM patients by indirect IFL that uses viable preparations of rat islet or insulinoma cells as substrate, but they also can be detected by using human insulinoma or fetal islet cells. We have tried to demonstrate ICSAs in the sera of 31 newly diagnosed diabetic patients, including 6 positive samples on human fetal islet cells, which used their natural target for the first time: normal human islet cells. In spite of using different types of preparations of these cells (i.e., freshly dispersed cell suspensions, monolayer cultures, or dispersed islets after culture), ICSAs could not be detected by IFL under the UV microscope, nor by flow cytometry. In contrast, 9 of 29 of the sera gave a positive staining on the RIN rat insulinoma cells. In an attempt to establish whether the putative ICSA autoantigen is present in the surface of human islet cells in the diabetic pancreas, the insulitis microenvironment was emulated by exposing the islets to three types of stress: 1) cytokines (IFN-gamma and TNF-alpha); 2) heat shock; and 3) hyperglycemia. However, diabetic sera failed again to recognize membrane antigens on the islet cells after either of these treatments. Neither were islet cells from a newly diagnosed diabetic patient stained by its autologous serum (ICA titer > 80 JDF U). These results suggest that ICSA autoantigen is not expressed in the membrane of human islet cells and therefore raises doubts about their proposed pathogenic role.


Mid-term survivorship of Mini-keel™ versus Standard keel in total knee replacements: Differences in the rate of revision for aseptic loosening.
To reduce the size of the surgical incision, modular mini-keel tibial components have been developed with or without extensions for the Nexgen™ MIS Tibial Component. Although a smaller component could theoretically result in defective fixation, this has never been evaluated in a large comparative series. Thus, we performed the following case control study to: (1) evaluate intermediate-term survival of a modular "mini-keel" tibial component compared to a reference standard keel component from the same line of products (Nexgen LPS-Flex Tibial Component, Zimmer); (2) to identify any eventual associated factors if the frequency of loosening was increased. The rate of revision for aseptic tibial loosening is comparable for both components. This comparative, retrospective, single center series of 459 consecutive total knee arthroplasties (TKA) was performed between 2007 and 2010: with 212 modular "mini-keel" (MK) tibial components and 247 "standard" (S) components. Survival, rate of revision for aseptic tibial loosening and identification of a radiolucent line were analyzed at the final follow-up. After a median follow-up of 5years, the rate of revision for tibial aseptic loosing was significantly higher in the MK group with 12 cases (5.7%) and 4 cases in the S group (1.6%) (P=0.036). The use of the MK component appears to be a prognostic factor for surgical revision (hazard ratio=3.86 (1.23-11.88), P=0.02) but not for the development of a radiolucent line (HR=1.75 (0.9-3.4), P=0.097). The mean delay before revision was 38months (8-64) in the MK group and 15.2months (8-22) in the S group (P=0.006). Individual factors, such as gender, body mass index (BMI) and pre- or postoperative alignment were not prognostic factors for revision or radiolucent lines. The modular "mini-keel" tibial component was associated with a greater risk of revision for tibial component loosening. Case control study, III.


Modeling the next generation sequencing read count data for DNA copy number variant study.
As one of the most recent advanced technologies developed for biomedical research, the next generation sequencing (NGS) technology has opened more opportunities for scientific discovery of genetic information. The NGS technology is particularly useful in elucidating a genome for the analysis of DNA copy number variants (CNVs). The study of CNVs is important as many genetic studies have led to the conclusion that cancer development, genetic disorders, and other diseases are usually relevant to CNVs on the genome. One way to analyze the NGS data for detecting boundaries of CNV regions on a chromosome or a genome is to phrase the problem as a statistical change point detection problem presented in the read count data. We therefore provide a statistical change point model to help detect CNVs using the NGS read count data. We use a Bayesian approach to incorporate possible parameter changes in the underlying distribution of the NGS read count data. Posterior probabilities for the change point inferences are derived. Extensive simulation studies have shown advantages of our proposed methods. The proposed methods are also applied to a publicly available lung cancer cell line NGS dataset, and CNV regions on this cell line are successfully identified.


Exploring structural features of the interaction between the scorpion toxinCnErg1 and ERG K+ channels.
The gamma-KTx-type scorpion toxins specific for K+ channels were found to interact with ERG channels on the turret region, while alpha-KTx3.2 Agitoxin-2 binds to the pore region of the Shaker K+ channel, and alpha-KTx5.3 BmP05 binds to the intermediate region of the small-conductance calcium-activated K-channel (SK(Ca)). In order to explore the critical residues for gamma-KTx binding, we determined the NMR structure of native gamma-KTx1.1 (CnErg1), a 42 amino acid residues scorpion toxin isolated from the venom of the Mexican scorpion Centruroïdes noxius Hoffmann, and we used computational evolutionary trace (ET) analysis to predict possible structural and functional features of interacting surfaces. The 1H-NMR three-dimensional solution structure of native ergtoxin (CnErg1) was solved using a total of 452 distance constraints, 13 3J(NH-Halpha) and 10 hydrogen bonds. The structure is characterized by 2 segments of alpha-helices and a triple-stranded antiparallel beta-sheet stabilized by 4 disulfide bridges. The ET and structural analysis provided indication of the presence of two important amino acid residue clusters, one hydrophobic and the other hydrophilic, that should be involved in the surface contact between the toxin and the channel. Some features of the proposed interacting surface are discussed.