Isolation and characterization of Bordetella pertussis phenotype variants capable of growing on nutrient agar: comparison with phases III and IV.
Unsupplemented nutrient agar (NA) was used to select spontaneous phenotype variants (PVs) of Bordetella pertussis Tohama I and 3779 which, by their growth on NA, could possibly be considered equivalent to phase IV in the system of Leslie and Gardner (P.H. Leslie and A.D. Gardner, J. Hyg. 31:423-434, 1931) or phase III in the system of Kasuga et al. (T. Kasuga, Y. Nakase, K. Ukishima, and K. Takatsu, Kitasato Arch. Exp. Med. 26:121-134, 1953). NA growers (Gna+) were selected from the flat, nonhemolytic, non-NA grower (Dom- Hly- Gna-) PV of both strains at a rate of between 10(-7) to 10(-8) per cell per generation. When cultured on Bordet-Gengou agar (BGA), more than one colony type was observed in strain 3779; these all retained the Gna+ characteristic during 10 to 30 passages on BGA. Analysis of 125I-surface-labeled whole cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed no major changes between the Dom- Hly- Gna+ PV and their Dom- Hly- Gna- PV parents in polypeptide profile (by Coomassie stain), in surface exposure of proteins (by autoradiography), or in lipopolysaccharide profile (by silver stain). Increased resistance to oleic acid, tetracycline, erythromycin, rifampin, and penicillin G, however, was characteristic for the Dom- Hly- Gna+ PV. Five phase IV strains and a phase III B. pertussis strain had similar antibiotic and oleic acid sensitivity profiles as the Dom- Hly- Gna+ isolates and plated with similar efficiency on NA, despite heterogeneity in BGA colonial morphology and lipopolysaccharide profile.


Musical expertise affects neural bases of letter recognition.
It is known that early music learning (playing of an instrument) modifies functional brain structure (both white and gray matter) and connectivity, especially callosal transfer, motor control/coordination and auditory processing. We compared visual processing of notes and words in 15 professional musicians and 15 controls by recording their synchronized bioelectrical activity (ERPs) in response to words and notes. We found that musical training in childhood (from age ~8 years) modifies neural mechanisms of word reading, whatever the genetic predisposition, which was unknown. While letter processing was strongly left-lateralized in controls, the fusiform (BA37) and inferior occipital gyri (BA18) were activated in both hemispheres in musicians for both word and music processing. The evidence that the neural mechanism of letter processing differed in musicians and controls (being absolutely bilateral in musicians) suggests that musical expertise modifies the neural mechanisms of letter reading.


Delayed diagnosis of large cerebello-pontine angle tumors, despite hearing conservation training.
Recently, at the National Naval Medical Center in Bethesda, Maryland, we have encountered several patients with large cerebello-pontine angle tumors. All were active duty and had evaded the military's stringent hearing screening program until the tumors were of life-threatening size. Early detection of these tumors can reduce overall morbidity and improve the patient's chances of returning to a normal lifestyle. We have analyzed each of these cases and discovered weaknesses in the program. The primary difficulty was in accurately following the algorithms set forth by military regulations. To that end we recommend: (1) establishing a cutoff for identification of those who should be evaluated after they obtain their enlistment/annual physical examinations; (2) specific training for hearing conservation program managers in the algorithms for hearing screening; (3) establishment of specific guidelines for primary care physicians for further evaluation of abnormal audiograms; and (4) reevaluation of form DD 2216.


Novel treatments for head and neck squamous cell carcinoma: preclinical identification and clinical investigation.
Head and neck squamous cell carcinoma (HNSCC) is a common cancer worldwide. Classically, it is a disease related to tobacco and alcohol use; an increasing number of patients are being diagnosed with HNSCC caused by infection with the human papillomavirus. New deep-sequencing techniques have confirmed the importance of p53 and EGF receptor in HNSCC development, and have identified pathways of critical importance, such as PI3K/mTOR and NOTCH. Increasing knowledge of key molecular features has lead to new therapeutic avenues for HNSCC. Novel therapies under investigation in HNSCC include antibody and small molecule inhibitors of EGF receptor and its family members, PI3K inhibitors, antiangiogenic agents, immunotherapies and agents interacting with early developmental pathways such as Hedgehog.


Sequence- and chain-length-specific complementary double-helix formation.
The artificial sequential strands consisting of two, three, or four m-terphenyl groups joined by diacetylene linkers with complementary binding sites, either the chiral amidine (A) or achiral carboxyl (C) group, were synthesized in a stepwise manner. Using circular dichroism and (1)H NMR spectroscopies along with liquid chromatography, we showed that, when three dimeric molecular strands (AA, CC, and AC) or six trimeric molecular strands (AAA, CCC, AAC, CCA, ACA, and CAC) were mixed in solution, the complementary strands were sequence-specifically hybridized to form one-handed double-helical dimers AA.CC and (AC) 2 or trimers AAA.CCC, AAC.CCA, and ACA.CAC, respectively, through complementary amidinium-carboxylate salt bridges. Upon the addition of CCA to a mixture of AAA, AAC, and ACA, the AAC.CCA double helix was selectively formed and then isolated from the mixture by chromatography. Moreover, the homo-oligomer mixtures of amidine or carboxylic acid from the monomers to tetramers (A, AA, AAAA, C, CC, and CCCC) assembled with a precise chain length specificity to form A.C, AA.CC, and AAAA.CCCC, which were separated by chromatography.


Testing and extending the triple match principle in the nursing profession: a generational perspective on job demands, job resources and strain at work.
The Triple Match Principle offers insight into the interactive interplay between job demands and job resources in the prediction of work-related strain. The aim of this article was to examine the interplay among job demands, job resources and strain in the nursing profession (the Triple Match Principle) and to gain insight into potential generational differences by investigating generation as a moderator of that interplay. No research has been done to evaluate generational differences in the Triple Match Principle. In a context of nursing shortages, it seems important to examine the relevance of the Triple Match Principle with respect to different generations of nurses. Cross-sectional study. A total of 1254 public healthcare sector nurses in Quebec, Canada, completed a questionnaire in the autumn of 2010. The questionnaire was used to assess cognitive, emotional and physical job demands and resources; psychological distress; psychosomatic complaints; and turnover intention. The results supported the Triple Match Principle and showed that job resources were more likely to buffer the effect of job demands on strain as the degree of match in qualitative dimension among demands, resources and strain increased (33·3% of triple-match interactions, 22·22% of double-match interactions and 16·67% non-match interactions were significant). Moreover, generation played a key role in this interplay, as it increased the number of significant qualitative interactions among job demands, job resources and strain. The results underscore the necessity of providing adequate job resources tailored to the specific job demands nurses face, to counteract the negative effects of those demands.


The α4β1/EMILIN1 interaction discloses a novel and unique integrin-ligand type of engagement.
EMILIN1, a homo-trimeric adhesive ECM glycoprotein, interacts with the α4β1 integrin through its gC1q domain. Uniquely among the C1q family members, the EMILIN1 gC1q presents only nine-stranded β-sandwich fold and the missing strand is substituted by a disordered 19-residue long segment spanning from Y927 to G945 at the apex of the gC1q domain. This unstructured loop exposes to the solvent the acidic residue E933, which plays a key role in the α4β1 integrin mediated interaction. Here, we experimentally determined that the three E933 residues (one from each monomer) are all required for ligand binding. By docking the NMR structure of the gC1q to a virtual α4β1 crystal structure based on the known structures of α4β7 and α5β1 integrins we built a model of α4β1-gC1q complex where three E933 residues are smoothly forced to coordinate the Mg2+ ion at the βI MIDAS site of the integrin. By bringing the three E933 close in space, the trimeric supramolecular organization of gC1q allows the formation of a proper 3D geometry and suggests a quaternary-structure-dependent mode of interaction. Furthermore, we experimentally identified R904 as a synergistic residue for cell adhesion. Accordingly, the model showed that this residue is able to form potential stabilizing intra-chain salt bridges with residues E928 and E930. This mode of interaction likely accounts for a more stable and durable α4β1-gC1q interaction in comparison with the prototypic CS1 ligand. To our knowledge, this is the first report describing the simultaneous involvement of all the three acidic residues of a trimeric ligand in the formation of a dimeric complex with the integrin βI domain.


New Findings on Unconsented Intimate Exams Suggest Racial Bias and Gender Parity.
Testimony from hundreds of medical students and numerous physicians and scholars suggests that unconsented intimate exams (UIEs) are unlikely to be rare, isolated incidents. However, much is unknown about the frequency of these exams and the circumstances in which they take place. The Community Bioethics Forum, founded and chaired by one of the authors of this commentary, is a consultative group of diverse community members who provide insights on law and policy to policy-makers and medical associations. Connecticut legislators asked the CBF to provide their views on proposed "explicit consent" legislation, and during those discussions, concerning narratives emerged about members' (and their loved ones') personal experiences with UIEs. To gain greater clarity on the demographic patterns and frequency of UIEs, we conducted the first national survey on UIEs. Data from this survey suggest that UIEs may occur under a broader range of circumstances than addressed by most law and policy. The survey resulted in nearly the exact same rate of affirmative responses between males and females in answer to whether they had received a UIE within the past five years. The survey results also showed evidence of racial disparity. Additional research is needed to understand the nature of UIEs.


Pilot randomized controlled trial of a hypnosis intervention for women with bladder pain syndrome.
To evaluate the feasibility and acceptability of a randomized controlled trial of a hypnosis intervention for the treatment of bladder pain syndrome/interstitial cystitis (BPS/IC) in women. We conducted a parallel arm, non-blinded, pilot randomized controlled trial of standardized hypnosis sessions including a hypnosis web tool versus usual care in adult women with BPS/IC. Pilot study outcomes included feasibility domains: process, resources and management, safety, and acceptability. Clinical outcomes of lower urinary tract symptoms and quality of life were measured using validated questionnaires at baseline and at the end of the 4-week intervention. We randomized 29 out of 30 (96.7%) eligible women. In the hypnosis group, 12 of 15 (80.0%) subjects completed the 4-week intervention and follow up, and 13 of 14 (92.9%) in the usual care group. In the hypnosis group, adherence to the standardized sessions was 80% and participants used the web-based tool for an average of 5.6 ± 2.7 times per week. Scores for emotional distress, relaxation, pain severity and expected bladder symptoms significantly improved during the first two of three planned hypnosis sessions (all p < 0.05). Improvement in quality of life scores was greater in the hypnosis group than the usual care group (-2.6 ± 2.3 vs. -0.9 ± 1.1, p = 0.04). There were no significant between-group differences in urinary symptoms or bladder pain. No adverse events were reported. A hypnosis intervention for the treatment of bladder pain syndrome/interstitial cystitis is feasible, acceptable, safe, and may improve quality of life.


HFE C282Y homozygotes have reduced low-density lipoprotein cholesterol: the Atherosclerosis Risk in Communities (ARIC) Study.
Recent studies have raised questions about the long-term health risks for individuals with mutations in the HFE gene, although previous studies may have been plagued by selection bias or lack of population-based comparison groups. We examined cardiovascular disease risk factors and iron and liver biomarkers, as well as morbidity and mortality associated with the C282Y and H63D variants of HFE in the Atherosclerosis Risk in Communities (ARIC) study, which is a population-based cohort of nearly 16,000 U.S. white and black men and women who were 45-64 years old at baseline. Subjects were followed for an average of 15 years for death, incident coronary heart disease, stroke, and heart failure, and an average of 8 years for incident diabetes. The prevalence of C282Y homozygosity was 0.42% (45/10,800) in whites, which is similar to other North American population-based studies. C282Y homozygotes had significantly lower mean low-density lipoprotein (LDL) cholesterol and fibrinogen as well as higher mean levels of iron (ferritin, transferrin saturation) and liver biomarkers (alanine aminotransferase, Hepascore) compared with HFE wild-type subjects. Rates of all-cause mortality, cardiovascular disease, and diabetes were similar across HFE genotypes. These prospective, population-based data indicate higher serum iron indices and possible mild liver dysfunction or disease in some C282Y homozygotes, but they provide little evidence that HFE C282Y or H63D mutations are related to all-cause mortality, cardiovascular disease, or diabetes. Reduced LDL in C282Y homozygotes may be because of effects of excess iron on cholesterol metabolism and lipoprotein formation in the liver.


Genes and loci involved in febrile seizures and related epilepsy syndromes.
Epilepsy is a paroxysmal disorder with a cumulative incidence of about 3%. About 13% of patients with epilepsy have a history of febrile seizures (FS). Generalized epilepsy with FS plus (GEFS+) is a familial epilepsy syndrome in which patients can have classic FS, FS that persist beyond the age of 5 years (i.e., FS+), and/or epilepsy. Both genetic and environmental factors have been shown to contribute to the pathogenesis of FS and GEFS+. During the past 10 years, molecular genetic studies have contributed a great deal to the identification of genetic factors involved in FS and GEFS+. In this study we aimed to provide a comprehensive review of currently known genes for FS and GEFS+, and the methods and approaches used to identify them. We also discuss the knowledge we currently have and hypotheses regarding the effect of the mutations on their respective protein functions.


Rural nurses' antiretroviral prescribing practices in children, Limpopo province, South Africa.
Errors in antiretroviral therapy (ART) use are common in children living with HIV (CLHIV), but there is limited evidence from rural primary healthcare (PHC) facilities where trained professional nurses initiate and manage most CLHIV. To assess antiretroviral prescribing practices of trained professional nurses in Mopani District's rural facilities and compare them to the 2015 national consolidated guidelines to evaluate the appropriateness of ART use. A four-year (2015-2018) retrospective cross-sectional medical record review was conducted of CLHIV in 94 rural PHC facilities of Mopani District. Inclusion criteria were: age under 15 years, initiated on ART by nurses in 2015 and virally unsuppressed (viral load ≥ 400 copies/mL) by the end of December 2018. A total of 16 669 antiretrovirals were prescribed from 7035 clinic visits. A correct ART regimen and dosage form was prescribed in 7045 (96%) and 15 502 (93%) of the cases. However, errors were common: 2928 (23%) incorrect doses, 15 502 (93%) incorrect dosing frequencies, and 4122 (61%) incorrectly dispensed antiretrovirals, and 3636 (28%) incorrect dosing frequencies. Antiretroviral prescribing errors in the form of drug omissions in ART regimens, incorrect dosing and dosing frequencies, lack of formulation considerations, and inadequate monthly supplies of antiretrovirals were commonly observed in this review. Antiretroviral stewardship programmes should be considered to develop and establish a fundamental strategy for improving quality in managing CLHIV.


PNA-functionalized magnetic microbeads as substrates for enzyme-labelled voltammetric genoassay for DNA sensing applied to identification of GMO in food.
A novel enzyme-labelled voltammetric magnetogenoassay for DNA sensing based on the use of carboxyl-surface coated magnetic microbeads functionalized with PNA probes and subsequent read-out on screen-printed electrode (SPE) substrates was developed. The assay was validated for determination of non-amplified genomic DNA from genetically modified Roundup Ready soy. Outstanding performance with respect to other genoassays requiring preliminary amplification of target DNA via PCR was demonstrated. The analytical performance was also improved compared to previous methods based on the immobilization of the same PNA probes on SPE substrates, since the method was found capable of achieving LOD and LOQ of 415 fM and 995 fM, respectively. The ability of the magnetogenoassay to detect the presence of Roundup Ready soy DNA sequence was tested on genomic DNA extract from European Reference Material soy flours, demonstrating the capability of the method to match the European Union regulation for labelling of food containing a percentage of GM products greater than 0,9%.


Dynamics of pneumococcal nasopharyngeal carriage in healthy children attending a day care center in northern Spain. Influence of detection techniques on the results.
Pneumococcal nasopharyngeal carriage precedes invasive infection and is the source for dissemination of the disease. Differences in sampling methodology, isolation or identification techniques, as well as the period (pre -or post-vaccination) when the study was performed, can influence the reported rates of colonization and the distribution of serotypes carried. To evaluate the prevalence and dynamics of pneumococcal nasopharyngeal colonization in healthy children aged 6-34 months attending a day care center with a high level of hygiene and no overcrowding. The study was performed 3-4 years after the 7-valent pneumococcal vaccine was introduced, using multiple methodologies to detect and characterize the isolates. Over 12 months, 25 children were sampled three times, 53 children twice and 27 children once. Three Streptococcus pneumoniae typing techniques were used: Quellung, Pneumotest-Latex-kit and multiplex-polymerase chain reaction (PCR). The similarity of isolates of the same serotype was established by pulsed field gel electrophoresis (PFGE) and occasionally the multilocus sequence type (ST) was also determined. Overall pneumococcal carriage and multiple colonization rates were 89.5% (94/105) and 39%, respectively. Among 218 pneumococci detected, 21 different serotypes and 13 non-typeable isolates were found. The most prevalent serotypes were 19A, 16F and 15B. Serotypes 15B, 19A and 21 were mainly found as single carriage; in contrast serotypes 6B, 11A and 20, as well as infrequent serotypes, were isolated mainly as part of multiple carriage. Most 19A isolates were ST193 but most serotypes showed high genetic heterogeneity. Changes in the pneumococci colonizing each child were frequent and the same serotype detected on two occasions frequently showed a different genotype. By multiplex-PCR, 100% of pneumococci could be detected and 94% could be serotyped versus 80.3% by the Quellung reaction and Pneumotest-Latex in combination (p < 0.001). Rates of S. pneumoniae carriage and multiple colonization were very high. Prevalent serotypes differed from those found in similar studies in the pre-vaccination period. In the same child, clearance of a pneumococcal strain and acquisition of a new one was frequent in a short period of time. The most effective technique for detecting pneumococcal nasopharyngeal carriers was multiplex-PCR.


Activation of protein kinase in the bovine corpus luteum by phospholipid and Ca2+.
A new species of protein kinase has been identified in cytosol preparations from bovine corpora lutea. Enzyme activity required the simultaneous presence of Ca2+ and phospholipid, and was also enhanced by glyceryl dioleate. Phosphatidylserine was the most effective phospholipid for stimulating histone phosphorylation. Other phospholipids capable of supporting enzymic activity were, in order of decreasing activity, phosphatidylinositol, phosphatidic acid, cardiolipin and phosphatidylglycerol. Several other phospholipids tested were ineffective. A cyclic AMP-dependent protein kinase was also present in the luteal cytosol. This enzyme activity was eliminated by protein kinase inhibitor without affecting the Ca2+- and phospholipid-stimulated activity. Lysine-rich histone (IIIS) was a much better substrate than type-IIA histone for Ca2+- and phospholipid-dependent phosphorylation. Ca2+ and phospholipid also enhanced phosphorylation of endogenous luteal cytosol protein. Calmodulin, alone or in the presence of Ca2+, was unable to increase phosphorylation. Trifluoperazine inhibited protein kinase activity stimulated by Ca2+ and phospholipid. These data suggest that a phospholipid-sensitive, Ca2+-dependent protein kinase may provide an important link between hormonally-induced changes in phospholipid metabolism and corpus-luteum function.


Absence of translationally selected synonymous codon usage bias in Helicobacter pylori.
Synonymous codon usage in the complete genome of Helicobacter pylori was investigated. The moderate A+T-richness of the genome (G+C=39 mol%) is reflected in the overall synonymous codon usage but the frequencies of some codons cannot be explained by simple mutational biases. A low level of heterogeneity among genes was observed, but this does not appear to be due to varying mutational bias or translational selection. Some of the heterogeneity was due to amino acid composition variation among the encoded proteins, and some may be attributable to recent acquisition of genes from other species. Since Hel. pylori codon usage is not dominated by biased mutation patterns, the absence of evidence for translationally mediated selection among synonymous codons is striking. This has implications with regard to the life history of this species, and in particular suggests that Hel. pylori strains are not subject to periods of competitive exponential growth. Despite the lack of selected codon usage, base composition immediately after the translation initiation site is skewed, consistent with selection against secondary structure formation in this region.


Dietary intake of vitamins B6, B12 and folate in relation to homocysteine serum concentration in the adult Polish population - WOBASZ Project.
Increased homocysteine (Hcy) levels predispose to atherosclerosis. Vitamin B6, B12 and folate play an important role in Hcy metabolism. To assess vitamin B6, B12 and folate intake and describe the relationship between these vitamins intake and Hcy level in the Polish population. Within the framework of the National Multicentre Health Survey (WOBASZ), a representative sample of the whole Polish population aged 20-74 was screened in the years 2003-2005. In each province of Poland 6 communities were randomly selected and in each of them a sample of 100 men and 100 women was randomly selected from the personal identification number (PESEL) database. In a sample of 50% of subjects (3004 men and 3401 women) Hcy level and nutrients were assessed according to the study protocol. Average intake of vitamin B6 was 2.26 mg/day in men and 2.03 mg/day in women, of vitamin B12 - 5.85 microg/day and 3.69 microg/day, and folate 258 microg/day and 211 microg/day, respectively. The recommended levels of intake were not achieved by 16% of men and 36% of women for vitamin B6, 32% and 51% for vitamin B12, and 78% and 90% for folate, respectively. The Hcy level (after adjustment for age, smoking, coffee and alcohol consumption) and prevalence of hyper-Hcy (> or = 12 micromol/l) decreased with increasing number of quartile of vitamins B6 and B12 in both genders and folate in men. In multivariable linear regression analysis an inverse association between the Hcy level and intake of vitamin B6 and folate in both genders, and vitamin B12 in men, was demonstrated. In the Polish population insufficient folate intake was common (deficiency was noted in nearly 80-90% of the population) and despite the correct average, a high proportion of subjects did not fulfil B6 and B12 intake recommendations; an inverse association between vitamins B6, B12 and folate consumption and Hcy concentration and prevalence of hyper-Hcy was observed


Transcribed and non-transcribed regions of Tetrahymena ribosomal gene chromatin have different accessibilities to micrococcal nuclease.
DNA renaturation kinetics was used to examine the relative accessibility of various regions of the Tetrahymena ribosomal RNA gene (rDNA) chromatin to micrococcal nuclease. In nuclei from cells active in rRNA transcription, the transcribed region of the rDNA chromatin was as much as 5-fold more accessible than the average of the total chromatin. As few as 20% inactive genes in the population could have accounted for all of the hybridization, so the transcribed region of the active units may be totally unprotected from nuclease degradation. The terminal non-transcribed spacer downstream from the transcription unit was also preferentially digested, but to a smaller degree. The central non-transcribed spacer was degraded to the same extent as total chromatin after a high degree of nuclease digestion. In nuclei from starved cells, which have 96% reduced rRNA transcription, the transcribed and terminal spacer regions of the rDNA were again more accessible than the total chromatin from the same nuclei, but the difference did not exceed 2-fold. We conclude that transcriptional activation is accompanied by major changes in the structure of the ribosomal gene chromatin, and that the extent and/or type of structural alteration differs in each functionally defined region of the rDNA.


Giving offspring a healthy start: parents' experiences of health promotion and lifestyle change during pregnancy and early parenthood.
There are good opportunities in Sweden for health promotion targeting expectant parents and parents of young children, as almost all are reached by antenatal and child health care. In 2005, a multisectoral child health promotion programme (the Salut Programme) was launched to further strengthen such efforts. Between June and December 2010 twenty-four in-depth interviews were conducted separately with first-time mothers and fathers when their child had reached 18 months of age. The aim was to explore their experiences of health promotion and lifestyle change during pregnancy and early parenthood. Qualitative manifest and latent content analysis was applied. Parents reported undertaking lifestyle changes to secure the health of the fetus during pregnancy, and in early parenthood to create a health-promoting environment for the child. Both women and men portrayed themselves as highly receptive to health messages regarding the effect of their lifestyle on fetal health, and they frequently mentioned risks related to tobacco and alcohol, as well as toxins and infectious agents in specific foods. However, health promotion strategies in pregnancy and early parenthood did not seem to influence parents to make lifestyle change primarily to promote their own health; a healthy lifestyle was simply perceived as 'common knowledge'. Although trust in health care was generally high, both women and men described some resistance to what they saw as preaching, or very directive counselling about healthy living and the lack of a holistic approach from health care providers. They also reported insufficient engagement with fathers in antenatal care and child health care. Perceptions about risks to the offspring's health appear to be the primary driving force for lifestyle change during pregnancy and early parenthood. However, as parents' motivation to prioritise their own health per se seems to be low during this period, future health promoting programmes need to take this into account. A more gender equal provision of health promotion to parents might increase men's involvement in lifestyle change. Furthermore, parents' ranking of major lifestyle risks to the fetus may not sufficiently reflect those that constitute greatest public health concern, an area for further study.


Effects of Recent Concussion and Injury History on Instantaneous Relative Risk of Lower Extremity Injury in Division I Collegiate Athletes.
Growing evidence suggests that concussion increases the risk of lower extremity (LE) musculoskeletal injury. However, it is unclear to how the effect of concussion on LE injury risk may be influenced by previous injuries. This study sought to examine the association between concussion, previous LE injuries, and the risk LE injury to the same previously injured limb (ipsilateral) or the opposite limb (contralateral). This retrospective study examined medical records from 110 concussed athletes and 110 matched controls for LE injuries in the 365 days before and after the concussion event. The effect of concussion on time to injury was assessed with a Cox proportional hazard model after adjusting for injury history. Fine and Gray subdistribution models assessed the cumulative risk of ipsilateral and contralateral injury by group. Concussion was associated with an increased instantaneous relative risk of LE injury when adjusting for LE injury history [hazard ratio (HR) = 1.67, 95% confidence interval (CI) = 1.11-2.53], agreeing with previous results. Among individuals who had a history of LE injuries before the concussion event, a nonsignificant yet moderate effect of concussion on the instantaneous relative risk of ipsilateral injuries was found after adjusting for the competing risk of contralateral injuries and censored values (HR = 1.85, 95% CI = 0.76-4.46). This study provides independent confirmation of previous studies, reporting an association between concussion and LE injury risk. Furthermore, this study suggests that future large-scale studies should consider the competing risk of ipsilateral, contralateral, and new injuries in populations with an injury history.


The expression of REG 1A and REG 1B is increased during acute amebic colitis.
Entamoeba histolytica, a protozoan parasite, is an important cause of diarrhea and colitis in the developing world. Amebic colitis is characterized by ulceration of the intestinal mucosa. We performed microarray analysis of intestinal biopsies during acute and convalescent amebiasis in order to identify genes potentially involved in tissue injury or repair. Colonic biopsy samples were obtained from 8 patients during acute E. histolytica colitis and again 60 days after recovery. Gene expression in the biopsies was evaluated using microarray, and confirmed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). REG 1A and REG 1B were the most up-regulated of all genes in the human intestine in acute versus convalescent E. histolytica disease: as determined by microarray, the levels of induction were 7.4-fold and 10.7 fold for REG 1A and B; p=0.003 and p=0.006 respectively. Increased expression of REG 1A and REG 1B protein in the colonic crypt epithelial cells during acute amebiasis was similarly observed by immunohistochemistry. Because REG 1 protein is anti-apoptotic and pro-proliferative, and since E. histolytica induces apoptosis of the intestinal epithelium as part of its disease process, we next tested if REG 1 might be protective during amebiasis by preventing parasite-induced apoptosis. Intestinal epithelial cells from REG 1-/- mice were found to be more susceptible to spontaneous, and parasite-induced, apoptosis in vitro (p=0.03). We concluded that REG 1A and REG 1B were upregulated during amebiasis and may function to protect the intestinal epithelium from parasite-induced apoptosis.


A comparison of two teaching simulations in preclinical operative dentistry.
The purpose of this study was to determine whether a ceramic tooth would be as effective as a natural tooth in teaching preclinical operative skills. Subjects consisted of all second-year students enrolled in a preclinical inlay course. Students were classified into high- and low-dexterity groups, randomly assigned to one of the tooth simulators, taught the same four procedures, and asked to keep a log of the amount of time spent on each. All students participated in a practical examination that consisted of preparing a cavity for a gold casting using a resin tooth. There were no statistically significant differences between students who prepared natural teeth and those who prepared ceramic teeth for either the qualitative practical examination score or the amount of preparation time required. High-dexterity students achieved significantly higher scores on the practical than did low-dexterity students, whether practicing on natural or ceramic teeth. The MOD onlay required significantly more time to complete than did any of the other preparations on both natural and ceramic tooth simulators. Written evaluations of the ceramic simulators revealed that students perceived ceramic teeth to be as good or better than natural teeth because of their anatomic uniformity.


The progression of vesicoureteral reflux nephropathy.
The relations among renal function, proteinuria, and glomerular lesions were studied in 54 patients with reflux nephropathy. The clinical course to end-stage renal disease was not appreciably altered by late surgical correction of the reflux, occurrence of urinary tract infection, or hypertension. All patients with progressive renal disease had significant proteinuria. Mesangial glomerular lesions can occur in the absence of proteinuria detectable by routine analysis, whereas lesions similar to those seen in idiopathic focal sclerosing glomerulopathy were present in the renal biopsies from proteinuric patients. Deposition of immunoproteins was limited to glomeruli undergoing sclerosis. Similarly, electron-dense deposits were confined to areas of mesangial alterations. Our results suggest that mesangial alterations occur early in the course of reflux nephropathy and may lead to the development of focal sclerosis. At later stages, counterproductive mechanisms of adaptation to the loss of viable nephrons might result in an acceleration of the clinical course to renal failure.


Vaccination against Strongyloides venezuelensis with homologue antigens using new immunomodulators.
Control of Strongyloides stercoralis infection is based on antihelminthic treatment. However, the development of an efficient vaccine is an important goal for more effectively managing this disease. Different Strongyloides spp. antigen preparations have been previously tested but without satisfactory results. In the present study, we evaluated whether the doxycholic acid extract of Strongyloides venezuelensis stage 3 larvae was able to protect CD1 mice against a homologue infection. Moreover, we included saponins from Quillaja saponaria (Qs) and immunomodulatory substances, i.e., Phlebodium pseudoaureum hydroalcoholic extract (PAL) or the amino alcohol AA0029, which has an aliphatic chain of 14 carbon atoms with a hydrophilic amino alcohol head, where the amino group is protected with a butoxycarbonyl group. The DOCSv antigen, together with the adjuvant components, was emulsified in a non-mineral water/oil emulsion. We performed duplicate experiments with each of the 2 immunomodulators. The DOCSv antigen used with Qs+PAL induced high levels of protection in terms of fecal egg count reduction (93.2-99.4%), 86-88% in adult worm recovery, and reduction in swelling of the small intestine. Mice vaccinated with the DOCSv antigen, together with Qs+AA0029, showed a 73.7-97.0% reduction in fecal egg count, with reduction in inflammation of the small intestine. High levels of IgG and IgG2a were observed in mice vaccinated with Qs+AA0029+DOCSv, indicating a Th1 immune response. Also, vaccinated mice recognized bands of 34, 39, 43, 95, and 170 kDa in a Western blot.


Towards Automated Infographic Design: Deep Learning-based Auto-Extraction of Extensible Timeline.
Designers need to consider not only perceptual effectiveness but also visual styles when creating an infographic. This process can be difficult and time consuming for professional designers, not to mention non-expert users, leading to the demand for automated infographics design. As a first step, we focus on timeline infographics, which have been widely used for centuries. We contribute an end-to-end approach that automatically extracts an extensible timeline template from a bitmap image. Our approach adopts a deconstruction and reconstruction paradigm. At the deconstruction stage, we propose a multi-task deep neural network that simultaneously parses two kinds of information from a bitmap timeline: 1) the global information, i.e., the representation, scale, layout, and orientation of the timeline, and 2) the local information, i.e., the location, category, and pixels of each visual element on the timeline. At the reconstruction stage, we propose a pipeline with three techniques, i.e., Non-Maximum Merging, Redundancy Recover, and DL GrabCut, to extract an extensible template from the infographic, by utilizing the deconstruction results. To evaluate the effectiveness of our approach, we synthesize a timeline dataset (4296 images) and collect a real-world timeline dataset (393 images) from the Internet. We first report quantitative evaluation results of our approach over the two datasets. Then, we present examples of automatically extracted templates and timelines automatically generated based on these templates to qualitatively demonstrate the performance. The results confirm that our approach can effectively extract extensible templates from real-world timeline infographics.


Important Details in Performing and Interpreting the Scratch Collapse Test.
The utility of the scratch collapse test has been demonstrated in examination of patients with carpal and cubital tunnel syndromes and long thoracic and peroneal nerve compressions. In the authors' clinic, this lesser known test plays a key role in peripheral nerve examination where localization of the nerve irritation or injury is not fully understood. Test utility and accuracy in patients with more challenging presentations likely correlate with tester understanding and experience. This article offers a clear outline of all stages of the test to improve interrater reliability. The nuances of test performance are described, including a description of situations where the scratch collapse test is deemed inappropriate. Four clinical scenarios where the scratch collapse test may be useful are included. Corresponding video content is provided to improve performance and interpretation of the scratch collapse test. Diagnostic, V.


Insights into the bovine rumen plasmidome.
Plasmids are self-replicating genetic elements capable of mobilization between different hosts. Plasmids often serve as mediators of lateral gene transfer, a process considered to be a strong and sculpting evolutionary force in microbial environments. Our aim was to characterize the overall plasmid population in the environment of the bovine rumen, which houses a complex and dense microbiota that holds enormous significance for humans. We developed a procedure for the isolation of total rumen plasmid DNA, termed rumen plasmidome, and subjected it to deep sequencing using the Illumina paired-end protocol and analysis using public and custom-made bioinformatics tools. A large number of plasmidome contigs aligned with plasmids of rumen bacteria isolated from different locations and at various time points, suggesting that not only the bacterial taxa, but also their plasmids, are defined by the ecological niche. The bacterial phylum distribution of the plasmidome was different from that of the rumen bacterial taxa. Nevertheless, both shared a dominance of the phyla Firmicutes, Bacteroidetes, and Proteobacteria. Evidently, the rumen plasmidome is of a highly mosaic nature that can cross phyla. Interestingly, when we compared the functional profile of the rumen plasmidome to two plasmid databases and two recently published rumen metagenomes, it became apparent that the rumen plasmidome codes for functions, which are enriched in the rumen ecological niche and could confer advantages to their hosts, suggesting that the functional profiles of mobile genetic elements are associated with their environment, as has been previously implied for viruses.


Desiccation tolerance in streptophyte algae and the algae to land plant transition: evolution of LEA and MIP protein families within the Viridiplantae.
The present review summarizes the effects of desiccation in streptophyte green algae, as numerous experimental studies have been performed over the past decade particularly in the early branching streptophyte Klebsormidium sp. and the late branching Zygnema circumcarinatum. The latter genus gives its name to the Zygenmatophyceae, the sister group to land plants. For both organisms, transcriptomic investigations of desiccation stress are available, and illustrate a high variability in the stress response depending on the conditions and the strains used. However, overall, the responses of both organisms to desiccation stress are very similar to that of land plants. We highlight the evolution of two highly regulated protein families, the late embryogenesis abundant (LEA) proteins and the major intrinsic protein (MIP) family. Chlorophytes and streptophytes encode LEA4 and LEA5, while LEA2 have so far only been found in streptophyte algae, indicating an evolutionary origin in this group. Within the MIP family, a high transcriptomic regulation of a tonoplast intrinsic protein (TIP) has been found for the first time outside the embryophytes in Z. circumcarinatum. The MIP family became more complex on the way to terrestrialization but simplified afterwards. These observations suggest a key role for water transport proteins in desiccation tolerance of streptophytes.


The "syringe" technique: a hands-free approach for the reduction of acute nontraumatic temporomandibular dislocations in the emergency department.
The traditional intraoral manual reduction of temporomandibular joint (TMJ) dislocations is time consuming, difficult, and at times ineffective, and commonly requires conscious sedation. We describe a novel technique for the reduction of acute nontraumatic TMJ dislocations in the emergency department (ED). This study was a prospective convenience sample population during a 3-year period at two university teaching-hospital EDs where acute nontraumatic TMJ dislocations were reduced utilizing our syringe technique. Demographics, mechanism, duration of dislocation, and reduction time were collected. Briefly, the "syringe" technique is a hands-free technique that requires a syringe to be placed between the posterior molars as they slide over the syringe to glide the anteriorly displaced condyle back into its normal anatomical position. Procedural sedation or intravenous analgesia is not required. Of the 31 patients, the mean age was 38 years. Thirty patients had a successful reduction (97%). The majority of dislocations were reduced in <1 min (77%). The two most common mechanisms for acute TMJ dislocations were due to chewing (n = 19; 61%) and yawning (n = 8; 29%). There were no recurrent dislocations at 3-day follow-up. We describe a novel technique for the reduction of the acutely nontraumatic TMJ dislocation in the ED. It is simple, fast, safe, and effective.


A Practice-Inspired Mindset for Researching the Psychophysiological and Medical Health Effects of Recreational Dance (Dance Sport).
"Dance" has been associated with many psychophysiological and medical health effects. However, varying definitions of what constitute "dance" have led to a rather heterogenous body of evidence about such potential effects, leaving the picture piecemeal at best. It remains unclear what exact parameters may be driving positive effects. We believe that this heterogeneity of evidence is partly due to a lack of a clear definition of dance for such empirical purposes. A differentiation is needed between (a) the effects on the individual when the activity of "dancing" is enjoyed as a dancer within different dance domains (e.g., professional/"high-art" type of dance, erotic dance, religious dance, club dancing, Dance Movement Therapy (DMT), and what is commonly known as hobby, recreational or social dance), and (b) the effects on the individual within these different domains, as a dancer of the different dance styles (solo dance, partnering dance, group dance; and all the different styles within these). Another separate category of dance engagement is, not as a dancer, but as a spectator of all of the above. "Watching dance" as part of an audience has its own set of psychophysiological and neurocognitive effects on the individual, and depends on the context where dance is witnessed. With the help of dance professionals, we first outline some different dance domains and dance styles, and outline aspects that differentiate them, and that may, therefore, cause differential empirical findings when compared regardless (e.g., amount of interpersonal contact, physical exertion, context, cognitive demand, type of movements, complexity of technique and ratio of choreography/improvisation). Then, we outline commonalities between all dance styles. We identify six basic components that are part of any dance practice, as part of a continuum, and review and discuss available research for each of them concerning the possible health and wellbeing effects of each of these components, and how they may relate to the psychophysiological and health effects that are reported for "dancing": (1) rhythm and music, (2) sociality, (3) technique and fitness, (4) connection and connectedness (self-intimation), (5) flow and mindfulness, (6) aesthetic emotions and imagination. Future research efforts might take into account the important differences between types of dance activities, as well as the six components, for a more targeted assessment of how "dancing" affects the human body.


Clinical Evaluation of the Safety, Efficacy and Tolerability of Lanthanum Carbonate in the Management of Hyperphosphatemia in Patients with End-Stage Renal Disease.
Patients with progressive chronic kidney disease (CKD) commonly develop mineral and bone abnormalities and extraskeletal calcifications with following increased cardiovascular risk. A key pathophysiological role is played by hyperphosphatemia. Since diet and dialysis are often insufficient to control serum phosphorus levels, many patients require treatment with phosphate binders. Among them is lanthanum carbonate, an aluminum-free non-calcium-based compound. The present review summarizes the most recent literature data concerning the safety, efficacy and tolerability of lanthanum carbonate in patients with end-stage renal disease and hyperphosphatemia. The drug is taken orally as chewable tablets or powder with only minimal gastrointestinal absorption and resulting reduced risk of tissue deposition and systemic drug interactions. The dissociation of the drug in the acid environment of the upper gastrointestinal tract induces the release of lanthanum ions, which bind to dietary phosphate forming insoluble complexes then excreted in the feces. Even though there is no clear evidence that lowering serum phosphorus levels can improve patient-centered outcomes, a mortality benefit with all phosphate binders, especially non-calcium containing ones, is not excluded. Lanthanum carbonate has been suggested to decrease all-cause mortality but not cardiovascular event rate compared to other phosphate binders. It induces a lower suppression of bone turnover than calcium carbonate and calcium acetate and may improve systolic function and cardiac dimension compared to calcium carbonate. Moreover, the use of lanthanum carbonate has been associated with better nutritional status compared to other phosphate binders, lower risk for hypercalcemia than calcium-containing binders, and amelioration of mild metabolic acidosis contrary to sevelamer hydrochloride. Main adverse effects include nausea, alkaline gastric reflux, gastric deposition of lanthanum, gastrointestinal obstruction, subileus, ileus, perforation, fecal impaction, and reduction of gastrointestinal absorption of some drugs including statins, angiotensin-converting enzyme inhibitors and some antibiotics such as fluoroquinolones or tetracyclines.


Noncanonical NF-κB activation mediates STAT3-stimulated IDO upregulation in myeloid-derived suppressor cells in breast cancer.
Immunotherapy for cancer treatment is achieved through the activation of competent immune effector cells and the inhibition of immunosuppressive cells, such as myeloid-derived suppressor cells (MDSCs). Although MDSCs have been shown to contribute to breast cancer development, the mechanism underlying MDSC-mediated immunosuppression is unclear. We have identified a poorly differentiated MDSC subset in breast cancer-suppressing T cell function through STAT3-dependent IDO upregulation. In this study we investigated the mechanisms underlying aberrant expression of IDO in MDSCs. MDSCs were induced by coculturing human CD33(+) myeloid progenitors with MDA-MB-231 breast cancer cells. Increased STAT3 activation in MDSCs was correlated with activation of the noncanonical NF-κB pathway, including increased NF-κB-inducing kinase (NIK) protein level, phosphorylation of cytoplasmic inhibitor of NF-κB kinase α and p100, and RelB-p52 nuclear translocation. Blocking STAT3 activation with the small molecule inhibitor JSI-124 significantly inhibited the accumulation of NIK and IDO expression in MDSCs. Knockdown of NIK in MDSCs suppressed IDO expression but not STAT3 activation. RelB-p52 dimers were found to directly bind to the IDO promoter, leading to IDO expression in MDSCs. IL-6 was found to stimulate STAT3-dependent, NF-κB-mediated IDO upregulation in MDSCs. Furthermore, significant positive correlation between the numbers of pSTAT3(+) MDSCs, IDO(+) MDSCs, and NIK(+) MDSCs was observed in human breast cancers. These results demonstrate a STAT3/NF-κB/IDO pathway in breast cancer-derived MDSCs, which provides insight into understanding immunosuppressive mechanisms of MDSCs in breast cancer.


Variational mean-field algorithm for efficient inference in large systems of stochastic differential equations.
This work introduces a Gaussian variational mean-field approximation for inference in dynamical systems which can be modeled by ordinary stochastic differential equations. This new approach allows one to express the variational free energy as a functional of the marginal moments of the approximating Gaussian process. A restriction of the moment equations to piecewise polynomial functions, over time, dramatically reduces the complexity of approximate inference for stochastic differential equation models and makes it comparable to that of discrete time hidden Markov models. The algorithm is demonstrated on state and parameter estimation for nonlinear problems with up to 1000 dimensional state vectors and compares the results empirically with various well-known inference methodologies.


Secondary structural features in the 70S RNAs of Moloney murine leukemia and Rous sarcoma viruses as observed by electron microscopy.
The secondary structural features in the 70S RNAs of the Prague strain of avian Rous sarcoma virus, subgroup A (PR-RSV-A), and Moloney murine leukemia virus (M-MuLV) were compared by electron microscopy. The PR-RSV-A genome contained two subunits joined by a linkage structure as in the genomes of M-MuLV and other mammalian retroviruses. In both viral genomes, a highly reproducible hairpin occurred at about 70 nucleotides from the 5' end of each subunit and contained 320 +/- 8 nucleotides. The stable point of linkage between the subunits in both viral genomes involved fewer than 50 nucleotides and occurred at 466 +/- 9 nucleotides from the 5' end. This places the linkage about 350 nucleotides further toward the 3' end of the subunit than the binding site of primer tRNA. Another structural feature common to both genomes was a loop in each subunit. In M-MuLV, the loop contained 3.9 +/- 0.10 kilobases (kb) and occurred at a distance of 2.2 +/- 0.05 kb from the 5' end. In PR-RSV-A, the loop was smaller (2.3 +/- 0.10 kb) and further (3.3 +/- 0.10 kb) from the 5' end. When M-MuLV RNA was heated to 70, 85, or 90 degrees C and cooled, the hairpin consistently reformed at the 5' end. No other structures typical of the native molecules reappeared. In RNA samples heated to 70 degrees C, a new loop reproducibly occurred near the 5' end of each subunit, but this loop was not found in samples heated to higher temperatures. Based on all of these findings, we conclude that the genome of PR-RSV-A shares several features with M-MuLV and other mammalian retroviruses and that the primer tRNA molecules are not involved in the linkage of the two subunits in either genome. We also conclude that the dimer linkage and the loops in subunits are typical of the native molecules and that their formation requires a special environment.


[The expression and clinical significance of Wnt-1 induced secreted protein-1 in breast carcinoma].
To investigate the expression of Wnt-1 induced secreted protein-1 (WISP-1) between breast cancer and paired normal breast tissues and to explore the significance of WISP-1 in breast cancer tumorigenesis. The mRNA and protein expressions of WISP-1 in human breast cancer were measured by Quantitative Real-Time RT-PCR and immunohistochemical staining and further analyzed the relationship between WISP-1 expression and clinic pathologic characters. WISP-1 expression in breast cancer was higher than that in normal breast tissue (P = 0.001). The mRNA expression level of WISP-1 was correlated with tumor size, staging, lymph node status, differentiated degree and HER-2 status (P < 0.05). WISP-1 protein expression level was correlated with lymph node status, differentiated degree and HER-2 status (P < 0.05). WISP-1 expression in human breast cancer increases significantly and may play a key role in the invasion and metastasis of human breast cancer.


Gaze pursuit and arm control of adolescent males diagnosed with attention deficit hyperactivity disorder (ADHD) and normal controls: evidence of a dissociation in processing visual information of short and long duration.
Three-dimensional kinematic analysis of line of gaze, arm and ball was used to describe the visual and motor behaviour of male adolescents diagnosed with attention deficit hyperactivity disorder (ADHD). The ADHD participants were tested when both on (ADHD-On) and off (ADHD-Off) their medication and compared to age-matched normal controls in a modified table tennis task that required tracking the ball and hitting to cued right and left targets. Long-duration information was provided by a pre-cue, in which the target was illuminated approximately 2 s before the serve, and short-duration information by an early-cue illuminated about 350 ms after the serve, leaving approximately 500 ms to select the target and perform the action. The ADHD groups differed significantly from the control group in both the pre-cue and early-cue conditions in being less accurate, in having a later onset and duration of pursuit tracking, and a higher frequency of gaze on and off the ball. The use of medication significantly reduced the gaze frequency of the ADHD participants, but surprisingly this did not lead to an increase in pursuit tracking, suggesting a barrier was reached beyond which ball flight information could not be processed. The control and ADHD groups did not differ in arm movement onset, duration and velocity in the short-duration early-cue condition; in the long-duration pre-cue condition, however, the ADHD group's movement time onset and arm velocity differed significantly from controls. The results show that the ADHD groups were able to process short-duration information without experiencing adverse effects on their motor behaviour; however, long-duration information contributed to irregular movement control.


Passion for Violently Themed Music and Psychological Well-Being: A Survey Analysis.
While the benefits to mood and well-being from passionate engagement with music are well-established, far less is known about the relationship between passion for explicitly violently themed music and psychological well-being. The present study employed the Dualistic Model of Passion to investigate whether harmonious passion (i.e., passionate engagement that is healthily balanced with other life activities) predicts positive music listening experiences and/or psychological well-being in fans of violently themed music. We also investigated whether obsessive passion (i.e., uncontrollable passionate engagement with an activity) predicts negative music listening experiences and/or psychological ill-being. Fans of violently themed music (N = 177) completed the passion scale, scale of positive and negative affective experiences, and various psychological well- and ill-being measures. As hypothesised, harmonious passion for violently themed music significantly predicted positive affective experiences which, in turn, predicted psychological well-being. Obsessive passion for violently themed music significantly predicted negative affective experiences which, in turn, predicted ill-being. Findings support the Dualistic Model of Passion, and suggest that even when music engagement includes violent content, adaptive outcomes are often experienced. We propose that the nature of one's passion for music is more influential in predicting well-being than the content or valence of the lyrical themes.


Crystal structure and nuclear magnetic resonance analyses of the SAND domain from glucocorticoid modulatory element binding protein-1 reveals deoxyribonucleic acid and zinc binding regions.
The glucocorticoid-modulatory element-binding proteins, GMEB1 and GMEB2, are ubiquitous, multifunctional DNA-binding proteins with important roles in the modulation of transcription upon steroid hormone activation. The GMEB proteins have intrinsic transactivation ability, but also control the glucocorticoid response via direct binding to the glucocorticoid receptor. They are also mandatory host proteins for Parvovirus replication. Here we present the 1.55 A resolution crystal structure of a central portion of GMEB1, encompassing its SAND domain, which shares 80% sequence identity with the GMEB2 SAND domain. We demonstrate that this domain, also present in numerous proteins implicated in chromatin-associated transcriptional regulation, is necessary and sufficient to bind the glucocorticoid-modulatory element (GME) DNA target. We use nuclear magnetic resonance (NMR) and binding studies to map the DNA recognition surface to an alpha-helical region exposing the conserved KDWK motif. Using site-directed mutagenesis, key residues for DNA binding are identified. In contrast to the previously determined NMR structure of the Sp100b SAND domain, we find that the GMEB1 SAND domain also comprises a zinc-binding motif. Although the zinc ion is not necessary for DNA binding, it is found to determine the C-terminal conformation of the GMEB1 SAND domain. We also show that homologous zinc-binding motifs exist in a subset of SAND domain proteins and probe the roles of this novel motif.


A distinctive patchy osteomalacia characterises Phospho1-deficient mice.
The phosphatase PHOSPHO1 is involved in the initiation of biomineralisation. Bones in Phospho1 knockout (KO) mice show histological osteomalacia with frequent bowing of long bones and spontaneous fractures: they contain less mineral, with smaller mineral crystals. However, the consequences of Phospho1 ablation on the microscale structure of bone are not yet fully elucidated. Tibias and femurs obtained from wild-type and Phospho1 null (KO) mice (25-32 weeks old) were embedded in PMMA, cut and polished to produce near longitudinal sections. Block surfaces were studied using 20 kV backscattered-electron (BSE) imaging, and again after iodine staining to reveal non-mineralised matrix and cellular components. For 3D characterisation, we used X-ray micro-tomography. Bones opened with carbide milling tools to expose endosteal surfaces were macerated using an alkaline bacterial pronase enzyme detergent, 5% hydrogen peroxide and 7% sodium hypochlorite solutions to produce 3D surfaces for study with 3D BSE scanning electron microscopy (SEM). Extensive regions of both compact cortical and trabecular bone matrix in Phospho1 KO mice contained no significant mineral and/or showed arrested mineralisation fronts, characterised by a failure in the fusion of the calcospherite-like, separately mineralising, individual micro-volumes within bone. Osteoclastic resorption of the uncalcified matrix in Phospho1 KO mice was attenuated compared with surrounding normally mineralised bone. The extent and position of this aberrant biomineralisation varied considerably between animals, contralateral limbs and anatomical sites. The most frequent manifestation lay, however, in the nearly complete failure of mineralisation in the bone surrounding the numerous transverse blood vessel canals in the cortices. In conclusion, SEM disclosed defective mineralising fronts and extensive patchy osteomalacia, which has previously not been recognised. These data further confirm the role of this phosphatase in physiological skeletal mineralisation.


Self-Reported Food Security in Adolescents with Type 1 Diabetes: Association with Hemoglobin A1c and Mental Health Symptoms Independent of Household Food Security.
Typically, child exposure to food insecurity is assessed by caregiver report of household food security. Child report has a potential for greater accuracy because it pertains only to the child whose experiences may differ from caregiver reports. We assessed if adolescent-reported food insecurity was associated with levels of hemoglobin A1c (HbA1c), acute diabetes-related complications, depressive symptoms, and disordered eating behaviors in adolescents with type 1 diabetes, independently from household food security. In a cross-sectional analysis of the multicenter SEARCH for Diabetes in Youth Study (Phase 4, 2016-2019) including 601 adolescents aged 10-17 years with type 1 diabetes and their caregivers, household food security and adolescent-reported food security were assessed using the 18-item Household Food Security Survey Module and the six-item Child Food Security Assessment questionnaire. Age-stratified (10-13, 14-17) regression models were performed to estimate independent associations, adjusting for socio-demographics, clinical factors, and household food security. Food insecurity was reported by 13.1% (n=79) of adolescents and 15.6% (n=94) of caregivers. Among adolescent-caregiver dyads, 82.5% (n=496) of reports were concordant and 17.5% (n=105) discordant, Cohen's kappa = 0.3. Adolescent-reported food insecurity was not independently associated with HbA1c, diabetic ketoacidosis, and severe hypoglycemia, including in age-stratified analyses. Adolescent-reported food insecurity was independently associated with elevated odds of depressive symptoms (Odds Ratio [OR] 3.6, 95% Confidence Interval [CI] 1.3-10.3) and disordered eating behaviors (OR 2.5, 95% CI 1.4-4.6) compared to adolescents reporting food security; these associations remained in both age groups for disordered eating behaviors and in the older group for depressive symptoms. Adolescents with type 1 diabetes may experience food insecurity differently than caregivers. Adolescent-reported food insecurity was independently associated with depressive symptoms and disordered eating behaviors and thus may be an important attribute to assess in addition to household food security in adolescents with type 1 diabetes.


Survival of enteroviruses in rapid-infiltration basins during the land application of wastewater.
The downward migration through soil of seeded poliovirus type 1 and echovirus type 1 and of naturally occurring enteroviruses during infiltration of sewage effluent through rapid-infiltration basins was investigated. After 5 days of flooding, the amount of seeded poliovirus type 1 that had migrated 5 to 10 cm downward through the soil profile was found to be 11% of that remaining at the initial burial depth. The amount of echovirus type 1 determined to have moved an equal distance was at least 100-fold less. Migration of naturally occurring enteroviruses during infiltration of sewage effluent through soil could not be measured with accuracy because of the possibility of virus survival from previous applications of effluent. The rate of inactivation for seeded poliovirus 1 and echovirus 1 buried in the infiltration basins ranged between 0.04 and 0.15 log10 units per day during the time when the basins were flooded. Inactivation of these same seeded virus types and of indigenous enterovirus populations in the infiltration basins during the drying portion of the sewage application cycle ranged between 0.11 and 0.52 log10 units per day. The rate of virus inactivation was dependent upon the rate of soil moisture loss. These results indicate that drying cycles during the land application of wastewater enhance virus inactivation in the soil.


Formin 1 and filamin B physically interact to coordinate chondrocyte proliferation and differentiation in the growth plate.
Filamin B (FlnB) is an actin-binding protein thought to transduce signals from various membrane receptors and intracellular proteins onto the actin cytoskeleton. Formin1 (Fmn1) is an actin-nucleating protein, implicated in actin assembly and intracellular signaling. Human mutations in FLNB cause several skeletal disorders associated with dwarfism and early bone fusion. Mouse mutations in Fmn1 cause aberrant fusion of carpal digits. We report here that FlnB and Fmn1 physically interact, are co-expressed in chondrocytes in the growth plate and share overlapping expression in the cell cytoplasm and nucleus. Loss of FlnB leads to a dramatic decrease in Fmn1 expression at the hypertrophic-to-ossification border. Loss of Fmn1-FlnB in mice leads to a more severe reduction in body size, weight and growth plate length, than observed in mice following knockout of either gene alone. Shortening of the long bone is associated with a decrease in chondrocyte proliferation and an overall delay in ossification in the double-knockout mice. In contrast to FlnB null, Fmn1 loss results in a decrease in the width of the prehypertrophic zone. Loss of both proteins, however, causes an overall decrease in the width of the proliferation zone and an increase in the differentiated hypertrophic zone. The current findings suggest that Fmn1 and FlnB have shared and independent functions. FlnB loss promotes prehypertrophic differentiation whereas Fmn1 leads to a delay. Both proteins, however, regulate chondrocyte proliferation, and FlnB may regulate Fmn1 function at the hypertrophic-to-ossification border, thereby explaining the overall delay in ossification.


Speed and accuracy of facial expression classification in avoidant personality disorder: a preliminary study.
The aim of this preliminary study was to examine whether individuals with avoidant personality disorder (APD) could be characterized by deficits in the classification of dynamically presented facial emotional expressions. Using a community sample of adults with APD (n = 17) and non-APD controls (n = 16), speed and accuracy of facial emotional expression recognition was investigated in a task that morphs facial expressions from neutral to prototypical expressions (Multi-Morph Facial Affect Recognition Task; Blair, Colledge, Murray, & Mitchell, 2001). Results indicated that individuals with APD were significantly more likely than controls to make errors when classifying fully expressed fear. However, no differences were found between groups in the speed to correctly classify facial emotional expressions. The findings are some of the first to investigate facial emotional processing in a sample of individuals with APD and point to an underlying deficit in processing social cues that may be involved in the maintenance of APD.


Effect of TGF-beta/Smad signaling pathway on lung myofibroblast differentiation.
Myofibroblasts play important roles in the pathogenesis of lung fibrosis. Transforming growth factor (TGF)-beta 1 has been widely recognized as a key fibrogenic cytokine. The major signaling pathway of (TGF)-beta(1) is through cytoplasmic Smad proteins. Our study investigated the role of individual (TGF)-beta(1)/Smad signal proteins in mediating alpha-smooth muscle actin (alpha-SMA) gene expression, which is a well-known key marker of myofibroblast differentiation. We transiently cotransfected alpha-SMA promoter-luciferase fusion plasmid (p895-Luc) and Smad expression plasmids and measured Luc activity in (TGF)-beta(1)-treated human fetal lung fibroblasts. We induced Smad3 knockout mice lung fibrosis by bleomycin. alpha-SMA protein expression was assessed by Western blotting. Collagen protein was analyzed by measuring hydroxyprolin. Myofibroblast morphology was assessed by immunohistochemistry. We found that the overexpression of Smad3, not Smad2 markedly increased (TGF)-beta(1)-induced alpha-SMA promoter activity and alpha-SMA protein expression in vitro, whereas the overexpression of dominant negative mutant Smad3 and Smad7 repressed (TGF)-beta(1)-induced alpha-SMA gene expression. Compared to wild-type mice, Smad3 knockout mice showed attenuated lung fibrosis after bleomycin treatment, manifested by lower collagen production and myofibroblast differentiation. Our study suggested (TGF)-beta(1)/Smad3 is a major pathway which regulated the myofibroblast differentiation. This result indicates a potential significance for future attempts of attenuating the progression of human lung fibrosis by the inhibition of the Smad3 cascade.


Pancreas transplantation: indications and consequences.
Pancreas transplantation continues to evolve as a strategy in the management of diabetes mellitus. The first combined pancreas-kidney transplant was reported in 1967, but pancreas transplant now represents a number of procedures, each with different indications, risks, benefits, and outcomes. This review will summarize these procedures, including their risks and outcomes in comparison to kidney transplantation alone, and how or if they affect the consequences of diabetes: hyperglycemia, hypoglycemia, and microvascular and macrovascular complications. In addition, the new risks introduced by immunosuppression will be reviewed, including infections, cancer, osteoporosis, reproductive function, and the impact of immunosuppression medications on blood pressure, lipids, and glucose tolerance. It is imperative that an endocrinologist remain involved in the care of the pancreas transplant recipient, even when glucose is normal, because of the myriad of issues encountered post transplant, including ongoing management of diabetic complications, prevention of bone loss, and screening for failure of the pancreas graft with reinstitution of treatment when indicated. Although long-term patient and graft survival have improved greatly after pancreas transplant, a multidisciplinary team is needed to maximize long-term quality, as well as quantity, of life for the pancreas transplant recipient.


Evaluating Flow Distribution in a Multiaquifer Recharge Well using an In-Situ Flowmeter.
Quantifying the flow rate distribution in a multiple-screen recharge well is relevant to understanding groundwater flow and solute transport behavior in managed aquifer recharge (MAR) operations. In this study, an impeller flowmeter was deployed to measure flow rate distribution in a multiple-screen MAR well under both recharge and pumping conditions screened in the multiple-strata of the Virginia Coastal Plain aquifer system. Preferential flow distribution in the well was observed through the uppermost screens during recharge while flow distribution was more evenly distributed along all screens under pumping conditions. Analysis of flow along individual screens also indicates preferential flow to the upper part of the screen during both recharge and pumping. Comparison of flowmeter results under both recharge and pumping conditions to previous site-specific measurements suggests that the distribution of flow may vary with time, depending on well screen condition and well rehabilitation efforts, and should be monitored over the duration of an MAR project. These results have implications for groundwater quality given that flow distribution in a multi-screen recharge well has profound impact on travel time and on transport modeling if flow is assumed to be steady and consistent under a range of operational conditions. This article is protected by copyright. All rights reserved.


Atypical trigeminal neuralgia: a consequence of central sensitization?
Trigeminal neuralgia (TN) is characterized by sudden, recurrent, usually unilateral, severe brief stabbing pains in the distribution of trigeminal nerve. Although it is widely accepted that blood vessel or tumor compression contributes to paroxysms of TN, the pathogenesis of persistent background pain in atypical TN patient is unclear. Central sensitization is pain hypersensitivity caused by central neural plasticity. It is responsible for many temporal and symptomatic features of acute and chronic pain. We hypothesize that central sensitization might account for some symptoms of atypical TN. Based on this hypothesis, we postulate that early medical intervention predicts good outcomes in TN and medicines which are effective on central sensitization may be potential agents for the treatment of atypical TN.


Enzyme-Assisted Fucoidan Extraction from Brown Macroalgae Fucus distichus subsp. evanescens and Saccharina latissima.
Fucoidans from brown macroalgae (brown seaweeds) have different structures and many interesting bioactivities. Fucoidans are classically extracted from brown seaweeds by hot acidic extraction. Here, we report a new targeted enzyme-assisted methodology for fucoidan extraction from brown seaweeds. This enzyme-assisted extraction protocol involves a one-step combined use of a commercial cellulase preparation (Cellic®CTec2) and an alginate lyase from Sphingomonas sp. (SALy), reaction at pH 6.0, 40 °C, removal of non-fucoidan polysaccharides by Ca2+ precipitation, and ethanol-precipitation of crude fucoidan. The workability of this method is demonstrated for fucoidan extraction from Fucus distichus subsp. evanescens (basionym Fucus evanescens) and Saccharina latissima as compared with mild acidic extraction. The crude fucoidans resulting directly from the enzyme-assisted method contained considerable amounts of low molecular weight alginate, but this residual alginate was effectively removed by an additional ion-exchange chromatographic step to yield pure fucoidans (as confirmed by 1H NMR). The fucoidan yields that were obtained by the enzymatic method were comparable to the chemically extracted yields for both F. evanescens and S. latissima, but the molecular sizes of the fucoidans were significantly larger with enzyme-assisted extraction. The molecular weight distribution of the fucoidan fractions was 400 to 800 kDa for F. evanescens and 300 to 800 kDa for S. latissima, whereas the molecular weights of the corresponding chemically extracted fucoidans from these seaweeds were 10-100 kDa and 50-100 kDa, respectively. Enzyme-assisted extraction represents a new gentle strategy for fucoidan extraction and it provides new opportunities for obtaining high yields of native fucoidan structures from brown macroalgae.


Pharmacokinetics of 5-azacitidine administered with phenylbutyrate in patients with refractory solid tumors or hematologic malignancies.
To characterize the pharmacokinetic behavior of 5-azacitidine (5-AC), a cytidine nucleoside analog, when given with phenylbutyrate, a histone deaceytlase inhibitor. Pharmacokinetic data were obtained from two trials involving patients with solid tumor and hematologic malignancies. 5-AC at doses ranging from 10 to 75 mg/m2/d was administered once daily as a subcutaneous injection for 5 to 21 days in combination with phenylbutyrate administered as a continuous intravenous infusion for varying dose and duration every 28 or 35 days. Serial plasma samples were collected up to 24 hours after 5-AC administration. 5-AC was quantitated using a validated liquid chromatograph/tandem mass spectrometry method. 5-AC was rapidly absorbed with the mean T(max) occurring at 0.47 hour. Average maximum concentration (C(max)) and area under the curve (AUC(0-infinity)) values increased in a dose-proportionate manner with increasing dose from 10 to 75 mg/m2/d; the mean +/- SD C(max) and AUC(0-infinity) at 10 mg/m2/d were 776 +/- 459 nM and 1,355 +/- 1,125 h*nM, respectively, and at 75 mg/m2/d were 4,871 +/- 1,398 nM and 6,582 +/- 2,560 h*nM, respectively. Despite a short terminal half-life of 1.5 +/- 2.3 hours, inhibition of DNA methyl transferase activity in tumors of patients receiving 5-AC has been documented. 5-AC is rapidly absorbed and eliminated when administered subcutaneously. Sufficient 5-AC exposure is achieved to produce pharmacodynamic effects in tumors.


[Clinical skills of Chilean general practitioners for the management of depressive disorders].
Diverse and sustained efforts have been developed to improve the management of depression by general practitioners (GPs), but they have not improved treatment coverage and quality of services. To explore the level of knowledge and clinical skills to diagnose and treat depression by GPs in Primary Health Care (PHC) in the Metropolitan Area of Santiago de Chile (RM). Theoretical knowledge (TK), diagnostic skills (DS) and treatment skills (TS) were evaluated in 56 GPs of the RM with a battery of specially designed instruments. In TK there were significant differences between GPs aged 31 years or less and their older counterparts and between Chilean and foreign doctors. Five percent of observed differences in TK were explained by age and nationality, respectively. Chilean GPs achieved higher scores in recognition of symptoms (RS), one of the dimensions of DS. No significant differences by age or nationality were observed for DS and TS. Achievement of GPs on tests measuring TK, DS, and TS was generally below 50%. This deficiency should be improved.


Lyapunov exponents in disordered chaotic systems: avoided crossing and level statistics.
The behavior of the Lyapunov exponents (LEs) of a disordered system consisting of mutually coupled chaotic maps with different parameters is studied. The LEs are demonstrated to exhibit avoided crossing and level repulsion, qualitatively similar to the behavior of energy levels in quantum chaos. Recent results for the coupling dependence of the LEs of two coupled chaotic systems are used to explain the phenomenon and to derive an approximate expression for the distribution functions of LE spacings. The depletion of the level spacing distribution is shown to be exponentially strong at small values. The results are interpreted in terms of the random matrix theory.


Single limb exercise: pilot study of physiological and functional responses to forced use of the hemiparetic lower extremity.
Stroke-related deficits can impede both functional performance and walking tolerance. Individuals with hemiparesis rely on the stronger limb during exercise and functional tasks. The single limb exercise (SLE) intervention was a unique training protocol that focused only on the hemiparetic limb. Our objective was to determine the effect of the SLE intervention on cardiorespiratory fitness parameters. Twelve participants (5 male) with a mean age of 60.6 +/- 14.5 years and 69.1 +/- 82.2 months post stroke participated in the training intervention. All participants performed SLE using the hemiparetic leg three times a week for 4 weeks. The nonhemiparetic limb served as the control limb and did not engage in SLE. Peak oxygen uptake (VO2 peak) and oxygen uptake (VO2) were measured at baseline and post intervention in all 12 participants. At pre and post intervention, gait velocity was assessed in a subset of participants (n = 7) using the 10-m fast-walk test. After the 4-week SLE training intervention, significant improvements were found for VO2 during submaximal work effort (P = .009) and gait velocity (n = 7) (P = .001). Peak oxygen uptake did not increase (P = .41) after the training intervention. These data suggest that SLE training was an effective method for improving oxygen uptake and reducing energy expenditure during submaximal effort. Unilateral exercise focused on the hemiparetic leg may be an effective intervention strategy to consider for stroke rehabilitation.


A case of fulminant liver failure associated with hepatitis C virus.
Fulminant hepatitis due to acute hepatitis C virus (HCV) infection is rarely observed. We present a case study of a 21-year-old male patient who developed HCV-associated fulminant hepatitis after receiving steroid pulse therapy for optic neuritis. Despite daily plasmapheresis, the disease progressed to irreversible liver failure with grade 3 hepatic encephalopathy by the 6th day after symptom onset. The patient received a liver transplant on the 8th day. Serum anti-HCV antibody was negative at that time, but became positive on the 12th day. Positive HCV RNA in the serum at admission was reported after transplantation. Positive changes in anti-HCV antibodies and acute hepatitis with massive necrosis in the histology of the explanted liver indicated fulminant hepatitis due to acute HCV infection. Because of severe hepatitis recurrence, he started 12 months of interferon therapy on the 48th day, and obtained sustained virological response. His anti-HCV antibodies became negative again by 1.5 years after cessation of therapy. HCV genomes recovered from the patient's serum on the 7th and 48th days revealed 2 different clones out of 20 clones with 30 % amino acid difference in hypervariable region 1 of HCV second envelope glycoprotein. One of the 2 clones expanded further after liver transplantation. We conclude that early diagnosis of HCV-associated fulminant hepatitis requires the detection of HCV RNA in the serum. Severe hepatitis recurrence after liver transplantation might occur in patients with fulminant hepatitis due to HCV because of its monoclonal expansion.


Do patients wish to be involved in decision making in the consultation? A cross sectional survey with video vignettes.
To determine patients' preferences for a shared or directed style of consultation in the decision making part of the general practice consultation. Structured interview, with video vignettes of acted consultations. 5 practices in Lothian, Scotland. 410 patients (adults and adults accompanying children) attending surgery appointments. Preference for shared or directed form of video vignette for five different presenting conditions. Patients varied in their preference for involvement in decision making in the consultation. Under multiple regression analysis, patients' preference was found to be independently predicted by the problem viewed (patients presented with physical problems preferred a directed approach), patients' age (patients aged 61 or older were more likely to prefer the directed approach), social class (social classes I and II were more likely to prefer the shared approach), and smoking status (smokers more likely to prefer the shared approach). Those patients who were able to answer (or who thought their doctor's style similar to those in the vignettes) were more likely to describe their own doctor's style as similar to their preferred style. No major association in preference was found with sex, frequency of attendance, or perceived chronic ill health. Patients may vary in their desire for involvement in decision making in consultations. Although this variation seems to depend on the presenting problem, age, social class, and smoking status, these associations are not absolute, with large minorities in each group. Doctors need the skills, knowledge of their patients, and the time to determine on which occasions, with which illnesses, and at which level their patients wish to be involved in decision making.


The fertility of negroes without southern rural experience: A re-examination of the 1960 GAF study findings with 1967 SEO Data.
Abstract A major finding of the 1960 'Growth of American Families' (GAF) study was that whites and blacks without southern rural experience had similar fertility. This paper reports on a re-examination of this finding with a substantially larger black sample. Data from the 1967 Survey of Economic Opportunity demonstrated that the residence background classification utilized in the GAF study defeated, in part, the attempt to remove the effects of rural experience on fertility. Indigenous urban blacks had 25% higher fertility than indigenous urban whites. The fertility of urban black migrants out of the rural South was sharply curtailed in contrast to those remaining in the rural South. Although urban blacks of southern rural background had nominally higher fertility than indigenous urban blacks, the difference was neither statistically nor substantively significant. These results demand a re-ordering of the interpretation of the impact that migration has on urban black fertility and the white-black differential in fertility.


Nurse staffing and unplanned extubation in the pediatric intensive care unit.
To determine the association between unplanned extubations and years of nurse experience and nurse-to-patient ratio in the pediatric intensive care unit (PICU). Case-control study. University-affiliated children's hospital PICU. Unplanned extubations were identified from January 1999 through December 2002. Three control patients for each of the patients experiencing an unplanned extubation were selected on three matching factors: age, intubation duration, and severity of illness as defined by the Pediatric Risk of Mortality (PRISM) III. None. Fifty-five of 1,004 intubated patients (5.5%) experienced an unplanned extubation during the 4-yr period. A conditional logistic regression analysis was used to evaluate the association between a patient's risk of an unplanned extubation and the nurse's years of PICU experience and nurse-to-patient ratio. Factors associated with unplanned extubations included the documentation of patient agitation (odds ratio, 2.99; 95% confidence interval, 1.14, 7.86) and a nurse-to-patient ratio of 1:2 (one nurse caring for two patients) relative to a nurse-to-patient ratio of 1:1 (odds ratio, 4.24; 95% confidence interval, 1.00, 19.10). Years of PICU nursing experience, patient restraints, and the method of sedation delivery (continuous infusion vs. intermittent bolus) were not associated with unplanned extubations. Pediatric patients are more likely to experience an unplanned extubation when being cared for by a nurse assigned to two patients compared with a nurse caring for one patient. To provide safe patient care, health care policymakers and hospital administrators should consider the nurse-to-patient ratio and its potential association with adverse events in hospitalized children.


Impact of copper sulfate on survival, behavior, midgut morphology, and antioxidant activity of Partamona helleri (Apidae: Meliponini).
Copper sulfate (CuSO4) is widely used in agriculture as a pesticide and foliar fertilizer. However, the possible environmental risks associated with CuSO4 use, particularly related to pollinating insects, have been poorly studied. In this study, we evaluated both lethal and sublethal effects of CuSO4 on the stingless bee Partamona helleri. Foragers were orally exposed to five concentrations of CuSO4 (5000, 1666.7, 554.2, 183.4, 58.4 μg mL-1), and the concentration killing 50% (LC50) was estimated. This concentration (142.95 μg mL-1) was subsequently used in behavioral, midgut morphology, and antioxidant activity analyses. Bee mortality increased with the ingestion of increasing concentrations of CuSO4. Ingestion at the estimated LC50 resulted in altered walking behavior and damage to the midgut epithelium and peritrophic matrix of bees. Furthermore, the LC50 increased the catalase or superoxide dismutase activities and levels of the lipid peroxidation biomarker malondialdehyde. Furthermore, the in situ detection of caspase-3 and LC3, proteins related to apoptosis and autophagy, respectively, revealed that these processes are intensified in the midgut of treated bees. These data show that the ingestion of CuSO4 can have considerable sublethal effects on the walking behavior and midgut of stingless bees, and therefore could pose potential risks to pollinators including native bees. Graphical abstract.


Cytokine activation patterns and biomarkers are influenced by microorganisms in community-acquired pneumonia.
The inflammatory response in community-acquired pneumonia (CAP) depends on the host and on the challenge of the causal microorganism. Here, we analyze the patterns of inflammatory cytokines, procalcitonin (PCT), and C-reactive protein (CRP) in order to determine their diagnostic value. This was a prospective study of 658 patients admitted with CAP. PCT and CRP were analyzed by immunoluminometric and immunoturbidimetric assays. Cytokines (tumor necrosis factor-α [TNF-α], IL-1β, IL-6, IL-8, and IL-10) were measured using enzyme immunoassay. The lowest medians of CRP, PCT, TNF-α, and IL-6 were found in CAP of unknown cause, and the highest were found in patients with positive blood cultures. Different cytokine profiles and biomarkers were found depending on cause: atypical bacteria (lower PCT and IL-6), viruses (lower PCT and higher IL-10), Enterobacteriaceae (higher IL-8), Streptococcus pneumoniae (high PCT), and Legionella pneumophila (higher CRP and TNF-α). PCT ≥ 0.36 mg/dL to predict positive blood cultures showed sensitivity of 85%, specificity of 42%, and negative predictive value (NPV) of 98%, whereas a cutoff of ≤ 0.5 mg/dL to predict viruses or atypicals vs bacteria showed sensitivity of 89%/81%, specificity of 68%/68%, positive predictive value of 12%/22%, and NPV of 99%/97%. In a multivariate Euclidean distance model, the lowest inflammatory expression was found in unknown cause and the highest was found in L pneumophila, S pneumoniae, and Enterobacteriaceae. Atypical bacteria exhibit an inflammatory pattern closer to that of viruses. Different inflammatory patterns elicited by different microorganisms may provide a useful tool for diagnosis. Recognizing these patterns provides additional information that may facilitate a broader understanding of host inflammatory response to microorganisms.


The effects of common genetic variants in oncogenes on ovarian cancer survival.
The 5-year survival rate for invasive epithelial ovarian cancer is <35%. It has been suggested that common, germline genetic variation may influence survival after cancer diagnoses, which might enable the prediction of response to treatment and survival in the clinical setting. The aim of this study was to evaluate associations between common germline genetic variants in the oncogenes BRAF, ERBB2, KRAS, NMI, and PIK3CA, and survival after a diagnosis of epithelial ovarian cancer. We evaluated the association between 34 tagging single nucleotide polymorphisms and survival in 1,480 cases of invasive epithelial ovarian cancer cases from three different studies. Cox regression analysis, stratified by study, was used to estimate per rare allele hazard ratios (HR). The minor allele rs6944385 in BRAF was significantly associated with poor survival [HR, 1.19; 95% confidence intervals (95% CI), 1.02-1.39; P = 0.024]. The association remained after adjusting for prognostic factors (adjusted HR, 1.20; 95 CI, 1.03-1.40; P = 0.018). A haplotype of BRAF was also associated with poor survival (HR, 1.24; 95% CI, 1.02-1.51; P = 0.029) and was more significant after adjustment (HR, 1.44; 95% CI, 1.15-1.81; P = 0.001). We also found evidence of an association between a KRAS haplotype and poor survival in serous subtype (HR, 1.69; 95% CI, 1.21-2.38; P = 0.002), but this was no longer significant after adjustment. Finally, when analyses were restricted to the serous histologic subtype, the rare allele rs10842513 in KRAS, was associated with poor survival (HR, 1.40; 95% CI, 1.10-1.78; P = 0.007). Common genetic variants in the BRAF and KRAS oncogenes may be important in the prediction of survival in patients with invasive epithelial ovarian cancer.


The second type II module from human matrix metalloproteinase 2: structure, function and dynamics.
Matrix metalloproteinase 2 (MMP-2, gelatinase A, 72 kDa type IV collagenase) has an important role in extracellular matrix degradation during cell migration and tissue remodeling. It is involved in development, inflammation, wound healing, tumor invasion, metastasis and other physiological and pathological processes. The enzyme cleaves several types of collagen, elastin, fibronectin and laminin. Binding to collagen is mediated by three repeats homologous to fibronectin type II modules, which are inserted in the catalytic domain in proximity to the active site. We have determined the NMR solution structure of the second type II module from human MMP-2 (col-2). The module exhibits a typical type II fold with two short double-stranded antiparallel beta sheets and three large loops packed around a cluster of conserved aromatic residues. Backbone amide dynamics, derived from (15)N relaxation experiments, correlate well with solvent accessibility and intramolecular hydrogen bonding. A synthetic peptide with the collagen consensus sequence, (Pro-Pro-Gly)(6), is shown to interact with the module. Spectral perturbations induced by (Pro-Pro-Gly)(6) binding reveal the region involved in the interaction of col-2 with collagen. The binding surface comprises exposed aromatic residues Phe21, Tyr38, Trp40, Tyr47, Tyr53 and Phe55, and the neighboring Gly33-Gly37 segment.


Impact of nationwide enhanced implementation of best practices in pancreatic cancer care (PACAP-1): a multicenter stepped-wedge cluster randomized controlled trial.
Pancreatic cancer has a very poor prognosis. Best practices for the use of chemotherapy, enzyme replacement therapy, and biliary drainage have been identified but their implementation in daily clinical practice is often suboptimal. We hypothesized that a nationwide program to enhance implementation of these best practices in pancreatic cancer care would improve survival and quality of life. PACAP-1 is a nationwide multicenter stepped-wedge cluster randomized controlled superiority trial. In a per-center stepwise and randomized manner, best practices in pancreatic cancer care regarding the use of (neo)adjuvant and palliative chemotherapy, pancreatic enzyme replacement therapy, and metal biliary stents are implemented in all 17 Dutch pancreatic centers and their regional referral networks during a 6-week initiation period. Per pancreatic center, one multidisciplinary team functions as reference for the other centers in the network. Key best practices were identified from the literature, 3 years of data from existing nationwide registries within the Dutch Pancreatic Cancer Project (PACAP), and national expert meetings. The best practices follow the Dutch guideline on pancreatic cancer and the current state of the literature, and can be executed within daily clinical practice. The implementation process includes monitoring, return visits, and provider feedback in combination with education and reminders. Patient outcomes and compliance are monitored within the PACAP registries. Primary outcome is 1-year overall survival (for all disease stages). Secondary outcomes include quality of life, 3- and 5-year overall survival, and guideline compliance. An improvement of 10% in 1-year overall survival is considered clinically relevant. A 25-month study duration was chosen, which provides 80% statistical power for a mortality reduction of 10.0% in the 17 pancreatic cancer centers, with a required sample size of 2142 patients, corresponding to a 6.6% mortality reduction and 4769 patients nationwide. The PACAP-1 trial is designed to evaluate whether a nationwide program for enhanced implementation of best practices in pancreatic cancer care can improve 1-year overall survival and quality of life. ClinicalTrials.gov, NCT03513705. Trial opened for accrual on 22th May 2018.