Establishing a linked sentinel surveillance system for blood-borne viruses and sexually transmissible infections: methods, system attributes and early findings.
To describe the attributes and key findings from implementation of a new blood-borne virus (BBV) and sexually transmissible infection (STI) sentinel surveillance system based on routine testing at clinical sites in Victoria, Australia. The Victorian Primary Care Network for Sentinel Surveillance (VPCNSS) on BBV and STI was established in 2006 at 17 sites. Target populations included men who have sex with men (MSM), young people and injecting drug users (IDU). Sites collected demographic and risk behaviour information electronically or using paper surveys from patients undergoing routine HIV or STI (syphilis, chlamydia (Chlamydia trachomatis)) or hepatitis C virus (HCV) testing. These data were linked with laboratory results. Between April 2006 and June 2008, data were received for 67 466 tests and 52 042 questionnaires. In clinics providing electronic data, >90% of individuals tested for HIV, syphilis and chlamydia had risk behaviour information collected. In other clinics, survey response rates were >85% (HIV), 43.5% (syphilis), 42.7-66.5% (chlamydia) and <20% (HCV). Data completeness was >85% for most core variables. Over time, HIV, syphilis and chlamydia testing increased in MSM, and chlamydia testing declined in females (P = 0.05). The proportion of positive tests among MSM was 1.9% for HIV and 2.1% for syphilis. Among 16-24-year-olds, the proportion positive for chlamydia was 10.7% in males and 6.9% in females. Among IDU, 19.4% of HCV tests were antibody positive. The VPCNSS has collected a large, rich dataset through which testing, risk behaviours and the proportion positive can be monitored in high-risk groups, offering a more comprehensive BBV and STI surveillance system for Victoria. Building system sustainability requires an ongoing focus.


Cystatin C modulates cerebral beta-amyloidosis.
The CST3 Thr25 allele of CST3, which encodes cystatin C, leads to reduced cystatin C secretion and conveys susceptibility to Alzheimer's disease. Here we show that overexpression of human cystatin C in brains of APP-transgenic mice reduces cerebral amyloid-beta deposition and that cystatin C binds amyloid-beta and inhibits its fibril formation. Our results suggest that cystatin C concentrations modulate cerebral amyloidosis risk and provide an opportunity for genetic risk assessment and therapeutic interventions.


The effect of treatment completion and length of stay on employment and crime in outpatient drug-free treatment.
Length of stay in treatment has been found to be a significant predictor of positive post-treatment outcomes, such as decreases in unemployment and crime. However, length of stay may be an incomplete predictor of successful treatment. Surprisingly, few studies have examined whether completing treatment in addition to length of stay is an important factor in explaining positive treatment outcomes. The objective of our study is to examine the effect that treatment completion and length of stay have on post-treatment employment and crime for patients in outpatient drug-free treatment, the largest treatment modality in the United States. We use conditional logit and multiple regression models with program-level indicator variables (fixed effects) to estimate the effect of treatment completion and length of stay on employment and crime controlling for drug use severity, previous treatment history, and other patient demographics. Data are from the National Treatment Improvement Evaluation Study and include 986 adults enrolled in outpatient drug-free programs across the United States. We find that treatment completion and length of stay are significantly related to post-treatment employment. Holding length of stay constant, the occurrence of employment at follow-up among patients who complete their planned treatment is almost 2 times that of patients who do not complete treatment. However, treatment completion did not have a statistically significant effect on the probability of post-treatment crime. Although our results are mixed, these findings suggest that greater attention should be placed on evaluating the importance of both length of stay and treatment completion in treatment outcome studies.


Colovesical fistula in sigmoid diverticulitis. A case report.
Colonic diverticulosis has continuously increased, noticeably left-sided disease. Colovesical fistula is an uncommon complication of diverticulitis, and its most common cause is diverticular disease. Pneumaturia and fecaluria are commonly related symptoms. We present the case of a 79-year-old woman complaining pneumaturia and fecaluria. Abdominal CT showed a colovesical fistula due to sigmoid diverticulitis. After surgical adhesiolysis between the sigmoid colon and the bladder, the defect of the bladder wall was repaired by simple closure. The colonic defect was treated by segmental resection including the rectosigmoid junction. Following the operation the patient continuously improved at months 6, 12 and 18 without evidence of recurrences.


An evaluation of combined narrative exposure therapy and physiotherapy for comorbid PTSD and chronic pain in torture survivors.
Torture is associated with adverse health consequences, with especially high rates of PTSD, depression and chronic pain. Despite increased awareness of the relationship between pain and posttraumaticsymptoms, and the accompanying need for effective treatment strategies, few studies have examined an integrated treatment of comorbid PTSD and pain. In this study, using an A-B case series design with three and six month follow-up, six refugee torture survivors with comorbid PTSD, depression and chronic pain received 20 sessions of Narrative Exposure Therapy (NET) and 10 sessions of physiotherapy. Outcome variables included symptoms of PTSD and depression, pain intensity, physical functioning and quality of life. Symptoms of PTSD and pain were also rated after each treatment session. Two patients achieved clinically significant reduction in symptoms of PTSD. Only one patient achieved clinically significant change in depressive symptoms, and two experienced clinically significant reduction in pain intensity. Clinical descriptions of the course of treatment for all patients are provided. Despite its limitations, the study suggests that some torture survivors who suffer high symptom loads may benefit from a combined treatment of NET and physiotherapy. Appreciating individual differences and how they affect treatment can provide valuable insight and inform clinicians working with torture survivors. Directions for future researchregarding the improvement of rehabilitation strategies of torture survivors are discussed, and highlighted through descriptions from the six therapy cases.


The 2019 Genitourinary Pathology Society (GUPS) White Paper on Contemporary Grading of Prostate Cancer.
Controversies and uncertainty persist in prostate cancer grading. To update grading recommendations. Critical review of the literature along with pathology and clinician surveys. Percent Gleason pattern 4 (%GP4) is as follows: (1) report %GP4 in needle biopsy with Grade Groups (GrGp) 2 and 3, and in needle biopsy on other parts (jars) of lower grade in cases with at least 1 part showing Gleason score (GS) 4 + 4 = 8; and (2) report %GP4: less than 5% or less than 10% and 10% increments thereafter. Tertiary grade patterns are as follows: (1) replace "tertiary grade pattern" in radical prostatectomy (RP) with "minor tertiary pattern 5 (TP5)," and only use in RP with GrGp 2 or 3 with less than 5% Gleason pattern 5; and (2) minor TP5 is noted along with the GS, with the GrGp based on the GS. Global score and magnetic resonance imaging (MRI)-targeted biopsies are as follows: (1) when multiple undesignated cores are taken from a single MRI-targeted lesion, an overall grade for that lesion is given as if all the involved cores were one long core; and (2) if providing a global score, when different scores are found in the standard and the MRI-targeted biopsy, give a single global score (factoring both the systematic standard and the MRI-targeted positive cores). Grade Groups are as follows: (1) Grade Groups (GrGp) is the terminology adopted by major world organizations; and (2) retain GS 3 + 5 = 8 in GrGp 4. Cribriform carcinoma is as follows: (1) report the presence or absence of cribriform glands in biopsy and RP with Gleason pattern 4 carcinoma. Intraductal carcinoma (IDC-P) is as follows: (1) report IDC-P in biopsy and RP; (2) use criteria based on dense cribriform glands (>50% of the gland is composed of epithelium relative to luminal spaces) and/or solid nests and/or marked pleomorphism/necrosis; (3) it is not necessary to perform basal cell immunostains on biopsy and RP to identify IDC-P if the results would not change the overall (highest) GS/GrGp part per case; (4) do not include IDC-P in determining the final GS/GrGp on biopsy and/or RP; and (5) "atypical intraductal proliferation (AIP)" is preferred for an intraductal proliferation of prostatic secretory cells which shows a greater degree of architectural complexity and/or cytological atypia than typical high-grade prostatic intraepithelial neoplasia, yet falling short of the strict diagnostic threshold for IDC-P. Molecular testing is as follows: (1) Ki67 is not ready for routine clinical use; (2) additional studies of active surveillance cohorts are needed to establish the utility of PTEN in this setting; and (3) dedicated studies of RNA-based assays in active surveillance populations are needed to substantiate the utility of these expensive tests in this setting. Artificial intelligence and novel grading schema are as follows: (1) incorporating reactive stromal grade, percent GP4, minor tertiary GP5, and cribriform/intraductal carcinoma are not ready for adoption in current practice.


Pulsed tissue Doppler identifies subclinical myocardial biventricular dysfunction in active acromegaly.
The aim of this study was to assess the role of pulsed tissue Doppler (TD) to identify left (LV) and right ventricular (RV) myocardial regional involvement in acromegaly. Thirty active acromegaly patients, free of diabetes mellitus, thyroid dysfunction, valvular and coronary heart disease, clinically overt heart failure, and 30 sex- and age-matched healthy controls underwent standard Doppler echocardiography and pulsed TD, by placing the sample volume at the level of basal posterior septum, LV lateral mitral annulus and RV lateral tricuspid annulus. Myocardial systolic (S(m)) and diastolic velocities (E(m)/A(m) ratio) and time-intervals of relaxation (RT(m)), precontraction (PCT(m)) and contraction (CT(m)) and the PCT(m)/CT(m) ratio were measured at each level. The two groups had similar heart rate, whereas acromegaly patients had higher body mass index, systolic and diastolic blood pressure, LV mass and impaired Doppler indexes of LV and RV diastolic function, without any difference in the global systolic function. At TD, acromegaly patients showed significantly delayed RT(m) and PCT(m,) reduced E(m)/A(m), S(m) and increased PCT(m)/CT(m) of posterior septum, mitral annulus and tricuspid annulus in comparison with controls. By separate multilinear regression analyses, after adjusting for body mass index, heart rate, diastolic blood pressure and LV mass index, age was the main independent determinant of tissue Doppler diastolic but not of systolic indexes. In active acromegaly, pulsed TD confirms LV and RV diastolic abnormalities detectable by standard Doppler, additionally identifying subclinical biventricular impairment of systolic function.


Prevalence of leisure-time sedentary behaviour and sociodemographic correlates: a cross-sectional study in Spanish adults.
Being physically inactive has been linked to a higher mortality and poorer quality of life. This cross-sectional study examines the prevalence of leisure-time sedentary behaviour in a population of Spanish adults and its correlates with several sociodemographic variables. Data were collected from 1,330 subjects living in Madrid (age: 18-65 years, 51.6% women) by telephone interview. The sampling error was ±2.7% for a 95.5% confidence level. Leisure-time sedentary behaviour was assessed using the Global Physical Activity Questionnaire (version 2). Further factors examined were: country of birth, sex, age, civil state, education level, employment and economic status and physical activity of parents. 76.3% of the subjects interviewed reported a mostly sedentary leisure-time lifestyle. The remaining subjects (23.7%) reported a moderate to high level of physical activity, meeting minimum recommendations. Logistic regression adjusted for all variables identified the following population subsets as showing a greater risk of sedentary behaviour: women (odds ratio (OR) = 2.14; 95% confidence interval (CI): 1.64, 2.79), participants aged 41-50 years (OR = 1.64; 95%CI:1.05, 2.51), those with a middle economic status (OR = 1.48; 95% CI: 1.04, 2.10) or not providing information about their income (OR = 1.97; 95% CI: 1.05, 3.67), and those whose father (OR = 1.53; 95% CI: 1.13, 2.07) and/or mother (OR = 1.41; 95% CI: 1.01, 1.97) were never physically active during leisure-time. The high prevalence of self-reported sedentary behaviour recorded suggests the need for public health policies targeted at increasing leisure-time physical activity levels. Our data identified several population subsets as priority candidates for possible interventions pursuing this goal.


Risk factors for delayed extubation after single-stage, multi-level anterior cervical decompression and posterior fusion.
Airway difficulties after single-stage, multilevel anterior and posterior cervical surgery are potentially life-threatening complications. Although extubation delays can occur, overnight intubation can reduce the risk of postoperative airway emergencies. Our protocol was as follows: all patients were kept intubated overnight in an intensive care unit and examined by the intensive care unit staff each morning. Readiness for extubation was based on the cuff-leak test, and extubation done on patients beyond the first postoperative day was considered delayed. Eleven patients were extubated on the first postoperative day (group 1), and 11 extubated beyond day 1 (group 2). No airway emergencies occurred. Patient factors-age, weight, smoking, medical comorbidities, American Society of Anesthesiologist class-were not significantly related to extubation delay. There were no differences between groups in the number of anterior and posterior levels or anterior and posterior operative times. Delayed extubation was significantly related to total operative time (8.2 hours vs. 10.6 hours), volume of crystalloid replacement (3,627 cm3 vs. 6,218 cm3) and intraoperative blood transfused (0.7 units vs. 3.1 units); approaching significance was increased blood loss (1,238 mL vs. 2,820 mL). We have found intraoperative factors-operative time, crystalloid volume, blood loss and replacement-rather than patient characteristics, to be risk factors for delayed extubation. Good communication with anesthesia staff and careful attention to postoperative airway management is essential after single-stage, multilevel anterior cervical decompression and posterior fusion.


Short- and long-term effects of individualized enteral protein supplementation in preterm newborns.
The aim of this retrospective study was to assess the need for additional enteral protein supplementation in preterm newborns with gestational age (GA) ≤32 weeks after full enteral feeds with either fortified breast milk (FBM) or preterm formula (PF) were reached, and to determine the effects of additional protein on physical and neurological development. After the standard early total parenteral nutrition (TPN) and reaching full enteral nutrition with 150-160 ml/kg/day, preterms were assessed for the requirement of additional protein based on serum blood urea nitrogen (BUN)/prealbumin levels. Additional enteral protein was given for BUN <5 mg/dl and/or prealbumin ≤8 mg/dl with weekly assessments as per Neonatal Intensive Care Unit (NICU) protocol. Growth in the NICU and neurodevelopmental outcome at 18 months' corrected age (CA) were determined. There were 32 newborns in the non-supplemented group (Group 1) and 33 newborns in the supplemented group (Group 2). All newborns in Group 2 were on FBM. Weight gain and head growth were better and Bayley scores at 18 months' CA were higher in Group 2. Standard preterm nutrition with FBM may not be sufficient for preterms, and additional enteral protein supplementation may improve the physical growth rate in the NICU and result in better neurodevelopmental outcome at 18 months' CA.


The forkhead transcription factor FoxI1 remains bound to condensed mitotic chromosomes and stably remodels chromatin structure.
All forkhead (Fox) proteins contain a highly conserved DNA binding domain whose structure is remarkably similar to the winged-helix structures of histones H1 and H5. Little is known about Fox protein binding in the context of higher-order chromatin structure in living cells. We created a stable cell line expressing FoxI1-green fluorescent protein (GFP) or FoxI1-V5 fusion proteins under control of the reverse tetracycline-controlled transactivator doxycycline inducible system and found that unlike most transcription factors, FoxI1 remains bound to the condensed chromosomes during mitosis. To isolate DNA fragments directly bound by the FoxI1 protein within living cells, we performed chromatin immunoprecipitation assays (ChIPs) with antibodies to either enhanced GFP or the V5 epitope and subcloned the FoxI1-enriched DNA fragments. Sequence analyses indicated that 88% (106/121) of ChIP sequences contain the consensus binding sites for all Fox proteins. Testing ChIP sequences with a quantitative DNase I hypersensitivity assay showed that FoxI1 created stable DNase I sensitivity changes in condensed chromosomes. The majority of ChIP targets and random targets increased in resistance to DNase I in FoxI1-expressing cells, but a small number of targets became more accessible to DNase I. Consistently, the accessibility of micrococcal nuclease to chromatin was generally inhibited. Micrococcal nuclease partial digestion generated a ladder in which all oligonucleosomes were slightly longer than those observed with the controls. On the basis of these findings, we propose that FoxI1 is capable of remodeling chromatin higher-order structure and can stably create site-specific changes in chromatin to either stably create or remove DNase I hypersensitive sites.


An evaluation of classification rules based on date of symptom onset to identify health-care associated infections.
The date of symptom onset is often used to distinguish health-care-associated from community-acquired infections. Those patients developing symptoms early in an inpatient stay are considered to have community-acquired infection, while those developing symptoms later are considered nosocomially infected. The authors evaluate the performance of this approach, showing how misclassification rates depend on the disease incubation period and the incidence rate ratio of infection among inpatients versus community members. The authors provide quantitative results for selecting classification rules that designate infections as health care associated or community acquired. These techniques allow the selection of disease-specific cutoffs to distinguish community- from nosocomially acquired infections that perform well for important illnesses. For example, a rule classifying those who develop flu symptoms in the first 1.5 days of their hospital stay as having community-acquired influenza and those developing symptoms later as having nosocomial infection has a positive predictive value and a negative predictive value of at least 87%. A cutoff of 6 days will identify community-acquired Legionnaires' disease with a positive predictive value and a negative predictive value of at least 77%. These results increase the utility of classifying infections by use of the date of onset by providing theoretically sound measures of performance, and they are applicable beyond the hospital setting.


CaMKIV regulates mitochondrial dynamics during sepsis.
Sepsis and shock states impose mitochondrial stress, and in response, adaptive mechanisms such as fission, fusion and mitophagy are induced to eliminate damaged portions of or entire dysfunctional mitochondria. The mechanisms underlying these events are being elucidated; yet a direct link between loss of mitochondrial membrane potential ΔΨm and the initiation of fission, fusion and mitophagy remains to be well characterized. The direct association between the magnitude of the ΔΨm and the capacity for mitochondria to buffer Ca2+ renders Ca2+ uniquely suited as the signal engaging these mechanisms in circumstances of mitochondrial stress that lower the ΔΨm. Herein, we show that the calcium/calmodulin-dependent protein kinase (CaMK) IV mediates an adaptive slowing in oxidative respiration that minimizes oxidative stress in the kidneys of mice subjected to either cecal ligation and puncture (CLP) sepsis or endotoxemia. CaMKIV shifts the balance towards mitochondrial fission and away from fusion by 1) directly phosphorylating an activating Serine616 on the fission protein DRP1 and 2) reducing the expression of the fusion proteins Mfn1/2 and OPA-1. CaMKIV, through its function as a direct PINK1 kinase and regulator of Parkin expression, also enables mitophagy. These data support that CaMKIV serves as a keystone linking mitochondrial stress with the adaptive mechanisms of mitochondrial fission, fusion and mitophagy that mitigate oxidative stress in the kidneys of mice responding to sepsis.


miR-520h is crucial for DAPK2 regulation and breast cancer progression.
MicroRNAs (miRNAs) are small RNAs that suppress gene expression by their interaction with 3'untranslated region of specific target mRNAs. Although the dysregulation of miRNAs has been identified in human cancer, only a few of these miRNAs have been functionally documented in breast cancer. Thus, defining the important miRNA and functional target involved in chemoresistance is an urgent need for human breast cancer treatment. In this study, we, for the first time, identified a key role of miRNA 520h (miR-520h) in drug resistance. Through protecting cells from paclitaxel-induced apoptosis, expression of miR-520h promoted the drug resistance of human breast cancer cells. Bioinformatics prediction, compensatory mutation and functional validation further confirmed the essential role of miR-520h-suppressed Death-associated protein kinase 2 (DAPK2) expression, as restoring DAPK2 abolished miR-520h-promoted drug resistance, and knockdown of DAPK2 mitigated cell death caused by the depletion of miR-520h. Furthermore, we observed that higher level of miR-520h is associated with poor prognosis and lymph node metastasis in human breast cancer patients. These results show that miR-520h is not only an independent prognostic factor, but is also a potential functional target for future applications in cancer therapeutics.


Fertility desires of Yoruba couples of South-western Nigeria.
Using the matched wife-husband (763) sample from the data collected from Ogbomoso and Iseyin towns in Oyo State, Nigeria, this paper examines factors associated with couples' fertility intention. The analysis used logistic regression models for predicting the effects of selected socioeconomic background characteristics on a couple's fertility intention. Results indicate high levels of concurrence among husbands and wives on fertility intention. Where differences exist, husbands are more pronatalists than their wives. About 87% of pairs of partners reported similar fertility preferences. Of these couples, 59.5% wanted more children while only 27.8% reported otherwise. The logistic regression models indicated that a couple's fertility intention was associated with age, education, place of residence, frequency of television-watching and number of living children. Therefore, programme interventions aimed at promoting fertility reduction in Nigeria should convey fertility regulation messages to both husbands and wives.


Replication of ribosomal DNA in Arabidopsis occurs both inside and outside the nucleolus during S phase progression.
Ribosomal RNA genes (rDNA) have been used as valuable experimental systems in numerous studies. Here, we focus on elucidating the spatiotemporal organisation of rDNA replication in Arabidopsis thaliana To determine the subnuclear distribution of rDNA and the progression of its replication during the S phase, we apply 5-ethynyl-2'-deoxyuridine (EdU) labelling, fluorescence-activated cell sorting, fluorescence in situ hybridization and structured illumination microscopy. We show that rDNA is replicated inside and outside the nucleolus, where active transcription occurs at the same time. Nascent rDNA shows a maximum of nucleolar associations during early S phase. In addition to EdU patterns typical for early or late S phase, we describe two intermediate EdU profiles characteristic for mid S phase. Moreover, the use of lines containing mutations in the chromatin assembly factor-1 gene fas1 and wild-type progeny of fas1xfas2 crosses depleted of inactive copies allows for selective observation of the replication pattern of active rDNA. High-resolution data are presented, revealing the culmination of replication in the mid S phase in the nucleolus and its vicinity. Taken together, our results provide a detailed snapshot of replication of active and inactive rDNA during S phase progression.


The enumeration and identification of acetic acid bacteria from South African red wine fermentations.
Acetic acid bacteria are microorganisms that can profoundly influence the quality of wine. Surprisingly, little research has been done on these microorganisms in the winemaking field. The object of this study was to investigate the occurrence of acetic acid bacteria in South African red wine fermentations and to identify the dominant species occurring. Acetic acid bacteria were isolated and enumerated from small-scale and commercial red must fermentations in 1998 and 1999, respectively. The initial occurrence of acetic acid bacteria in the must was shown to vary with cell numbers ranging from 10(6)-10(7) to 10(4)-10(5) cfu/ml for the 1998 and 1999 musts, respectively. The acetic acid bacteria decreased to 10(2)-10(3) cfu/ml in musts having a low pH (< or = 3.6), whereas in some musts having a high pH (> or = 3.7), the cell numbers increased during fermentation. During the process of cold soaking, the cell numbers of acetic acid bacteria also increased until inoculation with commercial wine yeast. Gluconobacter oxydans dominated in the fresh must and Acetobacter pasteurianus and A. liquefaciens during fermentation. This study showed that A. liquefaciens and A. hansenii were present in significant numbers, which has not been reported before.


Technical note: precision and accuracy of a commercially available CT optically stimulated luminescent dosimetry system for the measurement of CT dose index.
To determine the precision and accuracy of CTDI(100) measurements made using commercially available optically stimulated luminescent (OSL) dosimeters (Landaur, Inc.) as beam width, tube potential, and attenuating material were varied. One hundred forty OSL dosimeters were individually exposed to a single axial CT scan, either in air, a 16-cm (head), or 32-cm (body) CTDI phantom at both center and peripheral positions. Scans were performed using nominal total beam widths of 3.6, 6, 19.2, and 28.8 mm at 120 kV and 28.8 mm at 80 kV. Five measurements were made for each of 28 parameter combinations. Measurements were made under the same conditions using a 100-mm long CTDI ion chamber. Exposed OSL dosimeters were returned to the manufacturer, who reported dose to air (in mGy) as a function of distance along the probe, integrated dose, and CTDI(100). The mean precision averaged over 28 datasets containing five measurements each was 1.4% ± 0.6%, range = 0.6%-2.7% for OSL and 0.08% ± 0.06%, range = 0.02%-0.3% for ion chamber. The root mean square (RMS) percent differences between OSL and ion chamber CTDI(100) values were 13.8%, 6.4%, and 8.7% for in-air, head, and body measurements, respectively, with an overall RMS percent difference of 10.1%. OSL underestimated CTDI(100) relative to the ion chamber 21∕28 times (75%). After manual correction of the 80 kV measurements, the RMS percent differences between OSL and ion chamber measurements were 9.9% and 10.0% for 80 and 120 kV, respectively. Measurements of CTDI(100) with commercially available CT OSL dosimeters had a percent standard deviation of 1.4%. After energy-dependent correction factors were applied, the RMS percent difference in the measured CTDI(100) values was about 10%, with a tendency of OSL to underestimate CTDI relative to the ion chamber. Unlike ion chamber methods, however, OSL dosimeters allow measurement of the radiation dose profile.


Common DNA markers can account for more than half of the genetic influence on cognitive abilities.
For nearly a century, twin and adoption studies have yielded substantial estimates of heritability for cognitive abilities, although it has proved difficult for genomewide-association studies to identify the genetic variants that account for this heritability (i.e., the missing-heritability problem). However, a new approach, genomewide complex-trait analysis (GCTA), forgoes the identification of individual variants to estimate the total heritability captured by common DNA markers on genotyping arrays. In the same sample of 3,154 pairs of 12-year-old twins, we directly compared twin-study heritability estimates for cognitive abilities (language, verbal, nonverbal, and general) with GCTA estimates captured by 1.7 million DNA markers. We found that DNA markers tagged by the array accounted for .66 of the estimated heritability, reaffirming that cognitive abilities are heritable. Larger sample sizes alone will be sufficient to identify many of the genetic variants that influence cognitive abilities.


Popliteal-to-distal bypass: identifying risk factors associated with limb loss and graft failure.
Modern therapy, including endoluminal procedures and improved medical management, still yield less than desired results for tibial vessel occlusive disease. Despite the recent focus on these newer interventions, few modern series have evaluated the efficacy of popliteal-to-distal bypass procedures. The authors aimed to determine the efficacy of popliteal-distal bypass and to identify adverse prognostic factors for ultimate limb salvage. Eighty-seven patients (54 men; average age: 63 years) underwent 92 popliteal-distal bypasses. Duplex ultrasound was utilized to assess patency of all grafts. Data were analyzed by life-table analysis to determine patency rates at postoperative intervals. Median patient follow-up was 2.4 years. Major indications for bypass included chronic limb ischemia (86%) and disabling claudication (8%); 62% of the limbs were considered threatened, and 74% of the proximal anastomoses were above-knee. All procedures were technically successful. There were no perioperative (< 30 days) deaths, and 86% of patients were alive at 5 years. Cumulative patency rates were 74% at 6 months, 70% at 2 years, and 63% at 5 years. Limb salvage rates closely paralleled patency rates. At 5 years, 62% of the affected limbs were intact; 72% of the limbs lost were associated with early (< 180 days) bypass failures. Predictors of limb loss included early graft failure (84 days vs 1,288 days, p < 0.0001), younger age (57 years vs 64 years, p = 0.039), history of previous ipsilateral vascular procedures (50% vs 21%, p = 0.03), heavy (> 1 ppd) tobacco use (p = 0.001), and a thrombosed femoral-popliteal bypass at presentation (p = 0.002). When successful, popliteal-distal bypass is associated with excellent long-term patency and limb salvage rates. Early failures are often associated with limb loss. Heavy tobacco use, younger age, early graft failures, repeat revascularization, and presentation with a thrombosed femoral-popliteal graft are associated with limb loss.


A Rare Cause of Adrenal Insufficiency - Isolated ACTH Deficiency Due to TBX19 Mutation: Long-Term Follow-Up of Two Cases and Review of the Literature.
Isolated adrenocorticotropic hormone (ACTH) deficiency (IAD) is a rare cause of adrenal insufficiency and T-box pituitary restricted transcription factor (TBX19) mutations are responsible for two-thirds of the neonatal onset form of the disease. IAD presents with hypoglycemia and prolonged jaundice in the neonatal period. TBX19 is important for both pro-opiomelanocortin (POMC) gene transcription and differentiation of POMC-expressing cells. We describe 2 patients, 1 with a reported and 1 with a novel TBX19 mutation, and present information about the long-term follow-up of these patients. Both patients had critical illnesses, recurrent hypoglycemia, convulsions, and neonatal hyperbilirubinemia. They also had low cortisol and ACTH levels, while other pituitary hormones were within the normal range. Pituitary imaging was normal. After hydrocortisone treatment, there was resolution of the hypoglycemia and the convulsions were controlled. Genetic studies of the patients revealed both had inherited a homozygous mutation of the TBX19 gene. The first patient had an alteration of NM_005149.3:c.856C>T (p.R286*) and the second patient had a novel NM_005149.3:c.584C>T (p.T195I) mutation, analyzed by next-generation sequencing. The noteworthy findings of the patients at follow-up were: short stature, microcephaly, and decreased pubic hair in the first, and dysmorphic features, Chiari type 1 malformation, tall stature, and low bone mineral density (BMD) in the second. Congenital IAD can be life-threatening if it is not recognized and treated early. TBX19 mutations should be considered in the differential diagnosis of IAD. Further cases or functional analyses are needed for genotype-phenotype correlations. Low BMD, dysmorphic features, Chiari type 1 malformation, and sparse pubic hair are some of the important features in these patients.


A Biomechanical model of spiculated tumours under mammographic compressions.
The aim of this paper is to introduce effects well known to clinicians -but neglected to date- in the biomechanical modelling of breast malignant tumours. We develop a model of an isolated stellate breast tumour under mammographic compression forces. We study a range of reported mechanical properties, both linear elastic and hyperelastic. We also introduce different volumes of increased density/stiffness around the tumour. We show that each of these issues has a non-negligible effect on stresses and strains/deformations.


Quality improvement in cardiovascular surgery: results of a surgical quality improvement programme using a nationwide clinical database and database-driven site visits in Japan.
In 2015, an academic-led surgical quality improvement (QI) programme was initiated in Japan to use database information entered from 2013 to 2014 to identify institutions needing improvement, to which cardiovascular surgery experts were sent for site visits. Here, posthoc analyses were used to estimate the effectiveness of the QI programme in reducing surgical mortality (30-day and in-hospital mortality). Patients were selected from the Japan Cardiovascular Surgery Database, which includes almost all cardiovascular surgeries in Japan, if they underwent isolated coronary artery bypass graft (CABG), valve or thoracic aortic surgery from 2013 to 2016. Difference-in-difference methods based on a generalised estimating equation logistic regression model were used for pre-post comparison after adjustment for patient-level expected surgical mortality. In total, 238 778 patients (10 172 deaths) from 590 hospitals, including 3556 patients seen at 10 hospitals with site visits, were included from January 2013 to December 2016. Preprogramme, the crude surgical mortality for site visit and non-site visit institutions was 9.0% and 2.7%, respectively, for CABG surgery, 10.7% and 4.0%, respectively, for valve surgery and 20.7% and 7.5%, respectively, for aortic surgery. Postprogramme, moderate improvement was observed at site visit hospitals (3.6%, 9.6% and 18.8%, respectively). A difference-in-difference estimator showed significant improvement in CABG (0.29 (95% CI 0.15 to 0.54), p<0.001) and valve surgery (0.74 (0.55 to 1.00); p=0.047). Improvement was observed within 1 year for CABG surgery but was delayed for valve and aortic surgery. During the programme, institutions did not refrain from surgery. Combining traditional site visits with modern database methodologies effectively improved surgical mortality in Japan. These universal methods could be applied via a similar approach to contribute to achieving QI in surgery for many other procedures worldwide.


Principles of dermatoscopy of pigmented skin lesions.
There has been a dramatic increase in the incidence of malignant melanoma in most parts of the world. Because the tumor thickness is the most important prognostic factor for the prognosis of the malignant melanoma, the early detection of thin melanomas is essential. Dermatoscopy allows the physician to discriminate between melanocytic and nonmelanocytic lesions with high diagnostic accuracy and to detect initial malignant melanomas. We review the principles of dermatoscopy and the differential diagnosis of pigmented skin lesions. Before using the ABCD rule of dermatoscopy to classify melanocytic lesions into benign, suspicious, or malignant, the distinction between melanocytic and nonmelanocytic lesions is necessary. An essential prerequisite for the usefulness of this technique is adequate training.


A Process Evaluation of A UK Classroom-Based Physical Activity Intervention-'Busy Brain Breaks'.
The gap between development of effective physical activity interventions and the wide-scale adoption of these interventions in real-world settings has been reported since the early 2000s. Evaluations have been criticised for failing to report details of context, implementation, adoption and maintenance. 'Busy Brain Breaks' was an intervention designed to improve fundamental movement patterns whilst increasing physical activity within the classroom. This evaluation study used a mixed-methods approach including questionnaires, observations, semi-structured interviews and quantification of class-level dose. Findings suggest that giving teachers flexibility and autonomy over the way in which they implement physical activity interventions may increase the likelihood of adoption. Time was frequently perceived as a significant barrier to the intervention, giving the teachers flexibility to implement the intervention when they thought most suitable allowed teaching staff to retain their autonomy and make the intervention work with their schedule. Children's behaviour appeared to be both a facilitator and barrier to implementing physical activity interventions within the classroom. Whilst misbehaviour can pose as a barrier, children's enjoyment acts as a key facilitator to implementation for teaching practitioners. Teachers interviewed (n = 17) observed that movement ability had developed as a result of the intervention and recognised co-ordination, balance and stability as areas that had noticeably improved. Conducting an in-depth process evaluation has allowed for greater insight and understanding as to how, and to what extent, the intervention was implemented within the school-based setting.


DNA polymorphism haplotypes of the human apolipoprotein APOA1-APOC3-APOA4 gene cluster.
The genes coding for apolipoproteins A1, C3, and A4 (APOA1, APOC3, APOA4) are closely linked and tandemly organized within a 15-kilobase (kb) DNA segment on the long arm of human chromosome 11. The nucleotide variability of a 61-kb DNA segment containing these genes and their flanking sequences was studied by restriction analysis of a sample of 18 unrelated Northern Europeans using seven different genomic DNA probes. Eleven restriction site polymorphisms located within this DNA segment were used for haplotype analysis of 129 Mediterranean and 67 American black chromosomes. Estimation of the extent of nonrandom association between these polymorphisms indicated considerable linkage disequilibrium within the APOA1-APOC3-APOA4 gene cluster. Several haplotypes arose by recombination, and the rate of recombination within this gene cluster was estimated to be at least 4 times greater than that expected based on uniform recombination. The polymorphism information content of each of these polymorphisms, taken individually, ranges between 0.053 and 0.375, while that of their haplotypes ranges between 0.858 and 0.862. Therefore, DNA polymorphism haplotypes in the APOA1-APOC3-APOA4 gene cluster constitute a highly informative genetic marker on the long arm of human chromosome 11.


Characterization of a Spontaneously Immortalized Murine Müller Glial Cell Line QMMuC-1.
Müller glia are critical for the survival of retinal neurons and the integrity of retinal blood vessels. Müller glial cultures are important tools for investigating Müller glial pathophysiology. Here, we report a spontaneously immortalized Müller glial cell line originally cultured and subsequently cloned from mouse pups. The cell line, Queen's University Murine Müller glia Clone-1 (QMMuC-1), has been cultured for over 60 passages, has morphologic features like primary Müller cell (PMC) cultures and remains stable. QMMuC-1 and PMC cells were processed for immunohistochemistry, quantitative RT-PCR, Western blotting, whole cell voltage-clamping, and bioenergetic profiling. Immunocytochemistry showed that QMMuC-1 express known Müller glial markers, including glutamine synthetase, glial fibrillary acidic protein (GFAP), alpha-smooth muscle actin (α-SMA), Aquaporin 4, Kir4.1, interleukin 33 (IL-33), and sex determining region Y (SRY)-box2 (Sox2), but not Cone arrestin, Calbindin 1, CD68, and ionized calcium-binding adapter molecule 1 (Iba1). Compared with PMC, QMMuC-1 express higher levels of chemokine (C-C motif) ligand 2 (Ccl2), VEGFA, and glutamate aspartate transporter (GLAST), but lower levels of interleukin 6 (IL-6), brain-derived neurotrophic factor (BDNF), insulin-like growth factor 1 (IGF1), and neurotrophin 3 (NTF3). Whole-cell patch clamp recordings demonstrated characteristic inward currents in response to L-glutamate and L-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) by QMMuC-1 cells. The L-glutamate-induced current was significantly higher in QMMuC-1 cells compared with PMC. Bioenergetic profiling studies revealed similar levels of glycolysis and basal mitochondrial respiration between QMMuC-1 and PMC. However, mitochondrial spare capacity was significantly lower in QMMuC-1 compared with PMC. Our results suggest that the QMMuC-1 Müller glial cell line retains key characteristics of PMC with its unique profiles in cytokine/neurotrophic factor expression and mitochondrial respiration. QMMuC-1 has utility as an invaluable tool for understanding the role of Müller glia in physiological and pathological conditions.


The Risks to Patient Privacy from Publishing Data from Clinical Anesthesia Studies.
In this article, we consider the privacy implications of posting data from small, randomized trials, observational studies, or case series in anesthesia from a few (e.g., 1-3) hospitals. Prior to publishing such data as supplemental digital content, the authors remove attributes that could be used to re-identify individuals, a process known as "anonymization." Posting health information that has been properly "de-identified" is assumed to pose no risks to patient privacy. Yet, computer scientists have demonstrated that this assumption is flawed. We consider various realistic scenarios of how the publication of such data could lead to breaches of patient privacy. Several examples of successful privacy attacks are reviewed, as well as the methods used. We survey the latest models and methods from computer science for protecting health information and their application to posting data from small anesthesia studies. To illustrate the vulnerability of such published data, we calculate the "population uniqueness" for patients undergoing one or more surgical procedures using data from the State of Texas. For a patient selected uniformly at random, the probability that an adversary could match this patient's record to a unique record in the state external database was 42.8% (SE < 0.1%). Despite the 42.8% being an unacceptably high level of risk, it underestimates the risk for patients from smaller states or provinces. We propose an editorial policy that greatly reduces the likelihood of a privacy breach, while supporting the goal of transparency of the research process.


FAD2 Gene Radiation and Positive Selection Contributed to Polyacetylene Metabolism Evolution in Campanulids.
Polyacetylenes (PAs) are bioactive, specialized plant defense compounds produced by some species in the eudicot clade campanulids. Early steps of PA biosynthesis are catalyzed by Fatty Acid Desaturase 2 (FAD2). Canonical FAD2s catalyze desaturation, but divergent forms can catalyze hydroxylation, conjugation, acetylenation, and epoxygenation. These alternate reactions give rise to valuable unusual fatty acids, including the precursors to PAs. The extreme functional diversity of FAD2 enzymes and the origin of PA biosynthesis are poorly understood from an evolutionary perspective. We focus here on the evolution of the FAD2 gene family. We uncovered a core eudicot-wide gene duplication event giving rise to two lineages: FAD2-α and FAD2-β. Independent neofunctionalizations in both lineages have resulted in functionally diverse FAD2-LIKEs involved in unusual fatty acid biosynthesis. We found significantly accelerated rates of molecular evolution in FAD2-LIKEs and use this metric to provide a list of uncharacterized candidates for further exploration of FAD2 functional diversity. FAD2-α has expanded extensively in Asterales and Apiales, two main clades of campanulids, by ancient gene duplications. Here, we detected positive selection in both Asterales and Apiales lineages, which may have enabled the evolution of PA metabolism in campanulids. Together, these findings also imply that yet uncharacterized FAD2-α copies are involved in later steps of PA biosynthesis. This work establishes a robust phylogenetic framework in which to interpret functional data and to direct future research into the origin and evolution of PA metabolism.


Critical factors for EIA implementation: literature review and research options.
After decades of development, the gap between expectations of Environment Impact Assessments (EIA) and their practical performance remains significant. Research has been done to identify the critical factors for an effective implementation of EIA. However, this research, to a large extent, has not been cumulated and analysed comprehensively according to the stages of the EIA process. This paper contributes to the critical review of the literature on EIA implementation and effectiveness by cumulating mainly empirical findings in an implementation theoretical perspective. It focuses on the links between different critical factors and how they relate to different stages in the EIA and thus influence the decision making process. After reviewing 33 refereed journal articles published between 1999 and 2011, we identified 203 notions of critical factors. Of these, 102 related to different stages defined in our comprehensive EIA implementation model, and 101 were identified as general factors related to the whole EIA system. The number of notions of stage factors and general factors is thus about equal. An overlap between stage factors and general factors was found, which demonstrates that critical factors function differently in different cases. The function of the critical factors is complex and it is difficult to determine contingencies and causations. In the sources we examined, there is evidently an imbalance between in-depth empirical research and general knowledge, and the paper offers some suggestions for future research.


Child/Adolescent Anxiety Multimodal Study (CAMS): rationale, design, and methods.
To present the design, methods, and rationale of the Child/Adolescent Anxiety Multimodal Study (CAMS), a recently completed federally-funded, multi-site, randomized placebo-controlled trial that examined the relative efficacy of cognitive-behavior therapy (CBT), sertraline (SRT), and their combination (COMB) against pill placebo (PBO) for the treatment of separation anxiety disorder (SAD), generalized anxiety disorder (GAD) and social phobia (SoP) in children and adolescents. Following a brief review of the acute outcomes of the CAMS trial, as well as the psychosocial and pharmacologic treatment literature for pediatric anxiety disorders, the design and methods of the CAMS trial are described. CAMS was a six-year, six-site, randomized controlled trial. Four hundred eighty-eight (N = 488) children and adolescents (ages 7-17 years) with DSM-IV-TR diagnoses of SAD, GAD, or SoP were randomly assigned to one of four treatment conditions: CBT, SRT, COMB, or PBO. Assessments of anxiety symptoms, safety, and functional outcomes, as well as putative mediators and moderators of treatment response were completed in a multi-measure, multi-informant fashion. Manual-based therapies, trained clinicians and independent evaluators were used to ensure treatment and assessment fidelity. A multi-layered administrative structure with representation from all sites facilitated cross-site coordination of the entire trial, study protocols and quality assurance. CAMS offers a model for clinical trials methods applicable to psychosocial and psychopharmacological comparative treatment trials by using state-of-the-art methods and rigorous cross-site quality controls. CAMS also provided a large-scale examination of the relative and combined efficacy and safety of the best evidenced-based psychosocial (CBT) and pharmacologic (SSRI) treatments to date for the most commonly occurring pediatric anxiety disorders. Primary and secondary results of CAMS will hold important implications for informing practice-relevant decisions regarding the initial treatment of youth with anxiety disorders. ClinicalTrials.gov NCT00052078.


A comparison of physiological and transcriptome responses to water deprivation and salt loading in the rat supraoptic nucleus.
Salt loading (SL) and water deprivation (WD) are experimental challenges that are often used to study the osmotic circuitry of the brain. Central to this circuit is the supraoptic nucleus (SON) of the hypothalamus, which is responsible for the biosynthesis of the hormones, arginine vasopressin (AVP) and oxytocin (OXT), and their transport to terminals that reside in the posterior lobe of the pituitary. On osmotic challenge evoked by a change in blood volume or osmolality, the SON undergoes a function-related plasticity that creates an environment that allows for an appropriate hormone response. Here, we have described the impact of SL and WD compared with euhydrated (EU) controls in terms of drinking and eating behavior, body weight, and recorded physiological data including circulating hormone data and plasma and urine osmolality. We have also used microarrays to profile the transcriptome of the SON following SL and remined data from the SON that describes the transcriptome response to WD. From a list of 2,783 commonly regulated transcripts, we selected 20 genes for validation by qPCR. All of the 9 genes that have already been described as expressed or regulated in the SON by osmotic stimuli were confirmed in our models. Of the 11 novel genes, 5 were successfully validated while 6 were false discoveries.


Effectiveness of Workplace Interventions in Return-to-Work for Musculoskeletal, Pain-Related and Mental Health Conditions: An Update of the Evidence and Messages for Practitioners.
Purpose The objective of this systematic review was to synthesize evidence on the effectiveness of workplace-based return-to-work (RTW) interventions and work disability management (DM) interventions that assist workers with musculoskeletal (MSK) and pain-related conditions and mental health (MH) conditions with RTW. Methods We followed a systematic review process developed by the Institute for Work & Health and an adapted best evidence synthesis that ranked evidence as strong, moderate, limited, or insufficient. Results Seven electronic databases were searched from January 1990 until April 2015, yielding 8898 non-duplicate references. Evidence from 36 medium and high quality studies were synthesized on 12 different intervention categories across three broad domains: health-focused, service coordination, and work modification interventions. There was strong evidence that duration away from work from both MSK or pain-related conditions and MH conditions were significantly reduced by multi-domain interventions encompassing at least two of the three domains. There was moderate evidence that these multi-domain interventions had a positive impact on cost outcomes. There was strong evidence that cognitive behavioural therapy interventions that do not also include workplace modifications or service coordination components are not effective in helping workers with MH conditions in RTW. Evidence for the effectiveness of other single-domain interventions was mixed, with some studies reporting positive effects and others reporting no effects on lost time and work functioning. Conclusions While there is substantial research literature focused on RTW, there are only a small number of quality workplace-based RTW intervention studies that involve workers with MSK or pain-related conditions and MH conditions. We recommend implementing multi-domain interventions (i.e. with healthcare provision, service coordination, and work accommodation components) to help reduce lost time for MSK or pain-related conditions and MH conditions. Practitioners should also consider implementing these programs to help improve work functioning and reduce costs associated with work disability.


Treat-and-extend intravitreal bevacizumab for branch retinal vein occlusion.
To determine the effectiveness and direct medical costs of treat-and-extend (TAE) intravitreal bevacizumab (IVB) for the treatment of branch retinal vein occlusion (BRVO)-associated macular edema (ME). Retrospective chart review of 52 consecutive patients diagnosed with BRVO-associated ME treated with IVB using a TAE protocol. Mean change in logMAR vision was -0.30 (P < .001), and mean change in central macular thickness was -244.0 μm (P < .001). The mean number of injections was 8.2 (95% CI; 7.8 to 8.6). The yearly average direct cost of the TAE regimen was calculated to be $2,580.26 per patient. Treatment of BRVO-associated ME with IVB using a TAE regimen resulted in similar visual outcomes and number of intravitreal injections as did as-needed treatment with 0.5 mg ranibizumab conducted in phase 3 trials but with fewer visits and lower annual medical costs.


Correlations of maternal buprenorphine dose, buprenorphine, and metabolite concentrations in meconium with neonatal outcomes.
For the first time, relationships among maternal buprenorphine dose, meconium buprenorphine and metabolite concentrations, and neonatal outcomes are reported. Free and total buprenorphine and norbuprenorphine, nicotine, opiates, cocaine, benzodiazepines, and metabolites were quantified in meconium from 10 infants born to women who had received buprenorphine during pregnancy. Neither cumulative nor total third-trimester maternal buprenorphine dose predicted meconium concentrations or neonatal outcomes. Total buprenorphine meconium concentrations and buprenorphine/norbuprenorphine ratios were significantly related to neonatal abstinence syndrome (NAS) scores >4. As free buprenorphine concentration and percentage free buprenorphine increased, head circumference decreased. Thrice-weekly urine tests for opiates, cocaine, and benzodiazepines and self-reported smoking data from the mother were compared with data from analysis of the meconium to estimate in utero exposure. Time of last drug use and frequency of use during the third trimester were important factors associated with drug-positive meconium specimens. The results suggest that buprenorphine and metabolite concentrations in the meconium may predict the onset and frequency of NAS.


Safety and Outcome of Endovascular Treatment in Prestroke-Dependent Patients.
Background and Purpose- Prestroke dependence is an exclusion criterion in most trials of endovascular treatment (EVT) for acute ischemic stroke. Little is known about outcomes after EVT in these patients. We compared outcome and safety of EVT between prestroke-dependent and prestroke-independent patients. Methods- We report patients with an anterior circulation occlusion who were included between March 2014 and June 2016 in the MR CLEAN registry (Multicenter Randomized Clinical Trial of Endovascular Treatment of Acute Ischemic Stroke)-a prospective, multicenter, observational study for stroke intervention centers in the Netherlands. Prestroke dependence was defined as modified Rankin Scale score of 3 to 5 before onset of current stroke. Primary outcome was favorable outcome at 90 days, defined as modified Rankin Scale of 0 to 2 or not worsening of the modified Rankin Scale score. Secondary outcomes included National Institutes of Health Stroke Scale score post-intervention, reperfusion grade, and safety outcomes. Logistic regression analyses (adjusted for age, baseline National Institutes of Health Stroke Scale score, collaterals, time to EVT, and intravenous thrombolysis before EVT) were used to assess the association between prestroke dependence and outcomes. Results- One thousand four hundred forty-one patients were included in the present study, of whom 157 (11%) were prestroke dependent. Favorable outcome was seen in 27% prestroke-dependent patients, compared with 42% prestroke-independent patients ( P<0.05). After adjustment, prestroke dependence was not associated with less-favorable outcome (ORadjusted, 0.90; 95% CI, 0.58-1.39). The occurrence of symptomatic intracranial hemorrhage and ischemic stroke progression was similar in both groups. Conclusions- A substantial proportion of prestroke-dependent patients will reach prestroke modified Rankin Scale scores after EVT, and complication rates are comparable with prestroke-independent patients. Therefore, prestroke-dependent patients should not be routinely excluded from EVT.


Improvement of Local Cell Delivery Using Helix Transendocardial Delivery Catheter in a Porcine Heart.
Cardiac regeneration strategies using stem cells have shown variable and inconsistent results with respect to patient cardiac function and clinical outcomes. There has been increasing consensus that improving the efficiency of delivery may improve results. The Helix transendocardial delivery system (BioCardia Inc.) has been developed to enable percutaneous transendocardial biotherapeutic delivery. Therefore, we evaluated cell retention using this unique system compared with direct transepicardial injection and intracoronary infusion in an animal model.Twelve healthy swine were used in this study. 18Fluorodeoxyglucose (FDG)-labeled bone marrow mononuclear cells were delivered via percutaneous transendocardial route using the Helix system (TE group, n = 5), via direct transepicardial injection using a straight 27-gauge needle in an open chest procedure (TP group, n = 4), or via percutaneous intracoronary (IC) infusion (IC group, n = 3). One hour after cell delivery, the distribution of injected cells within the myocardium was assessed by PET-CT. Regions of interest were defined and their signals were compared in each group. Retention rates were calculated as a percentage of the comparing signal.The distribution of injected cells in the myocardium was higher in the TE group (17.9%) than in the TP group (6.0%, versus TE, P < 0.001) and the IC group (1.0%, versus TE, P < 0.001). Consistent with previous reports, there were signal distributions in the lungs, liver, and kidneys in qualitative whole body PET assessment.TE cell delivery using a helical infusion catheter is more efficient in cell retention than either TP delivery or IC delivery using PET-CT analysis.


A single-center randomized, controlled trial investigating the efficacy of a mHealth ECG technology intervention to improve the detection of atrial fibrillation: the iHEART study protocol.
Atrial fibrillation is a major public health problem and is the most common cardiac arrhythmia, affecting an estimated 2.7 million Americans. The true prevalence of atrial fibrillation is likely underestimated because episodes are often sporadic; therefore, it is challenging to detect and record an occurrence in a "real world" setting. To date, mobile health tools that promote earlier detection and treatment of atrial fibrillation and improvement in self-management behaviors and knowledge have not been evaluated. This study will be the first to address the epidemic problem of atrial fibrillation with a novel approach utilizing advancements in mobile health electrocardiogram technology to empower patients to actively engage in their healthcare and to evaluate impact on quality of life and quality-adjusted life years. Furthermore, sending a daily electrocardiogram transmission, coupled with receiving educational and motivational text messages aimed at promoting self-management and a healthy lifestyle may improve the management of chronic cardiovascular conditions (e.g., hypertension, diabetes, heart failure, etc.). Therefore, we are currently conducting a randomized controlled trial to assess the efficacy of a mobile health intervention, iPhone® Helping Evaluate Atrial fibrillation Rhythm through Technology (iHEART) versus usual cardiac care. The iHEART study is a single center, prospective, randomized controlled trial. A total of 300 participants with a recent history of atrial fibrillation will be enrolled. Participants will be randomized 1:1 to receive the iHEART intervention, receiving an iPhone® equipped with an AliveCor® Mobile ECG and accompanying Kardia application and behavioral altering motivational text messages or usual cardiac care for 6 months. This will be the first study to investigate the utility of a mobile health intervention in a "real world" setting. We will evaluate the ability of the iHEART intervention to improve the detection and treatment of recurrent atrial fibrillation and assess the intervention's impact on improving clinical outcomes, quality of life, quality-adjusted life-years and disease-specific knowledge. NCT02731326 ; Verified April 2016.


Targeted hepatitis C antibody testing interventions: a systematic review and meta-analysis.
Testing for hepatitis C virus (HCV) infection may reduce the risk of liver-related morbidity, by facilitating earlier access to treatment and care. This review investigated the effectiveness of targeted testing interventions on HCV case detection, treatment uptake, and prevention of liver-related morbidity. A literature search identified studies published up to 2013 that compared a targeted HCV testing intervention (targeting individuals or groups at increased risk of HCV) with no targeted intervention, and results were synthesised using meta-analysis. Exposure to a targeted testing intervention, compared to no targeted intervention, was associated with increased cases detected [number of studies (n) = 14; pooled relative risk (RR) 1.7, 95% CI 1.3, 2.2] and patients commencing therapy (n = 4; RR 3.3, 95% CI 1.1, 10.0). Practitioner-based interventions increased test uptake and cases detected (n = 12; RR 3.5, 95% CI 2.5, 4.8; and n = 10; RR 2.2, 95% CI 1.4, 3.5, respectively), whereas media/information-based interventions were less effective (n = 4; RR 1.5, 95% CI 0.7, 3.0; and n = 4; RR 1.3, 95% CI 1.0, 1.6, respectively). This meta-analysis provides for the first time a quantitative assessment of targeted HCV testing interventions, demonstrating that these strategies were effective in diagnosing cases and increasing treatment uptake. Strategies involving practitioner-based interventions yielded the most favourable outcomes. It is recommended that testing should be targeted at and offered to individuals who are part of a population with high HCV prevalence, or who have a history of HCV risk behaviour.


Conditions requiring hospitalisations, more than general anaesthesia itself, are associated with diagnosis of learning disorders in children.
Anaesthesia is neurotoxic in developing primates. Retrospective clinical studies show a correlation between exposure to anaesthesia during infancy and the occurrence of learning disorders (LD). Prospective studies failed to detect any influence of a single exposure to anaesthesia on neurodevelopment. We hypothesised that some specific populations of children were electively sensitive to anaesthesia-related neurotoxicity. Using a case-control design, we analysed the medical histories of children with LD, compared to those of their normally reading siblings. Interviews were conducted and medical records were reviewed. The numbers of hospitalisations and anaesthesia exposures before the age of five years were determined. Four hundred fourteen dyslexic children were screened over a one-year period. Two hundred and seventy patients were excluded due to confounding variables (single child, all siblings showing LD or any condition placing the neurological prognosis at risk (N = 107/414 for the latter)) or inability to accurately collect evaluation criteria. In the 144 case-control pairs studied, the mean number of hospitalisations was significantly different (N = 1.097 ± 0 .135/case versus 0.667 ± 0.097/control, p = 0.0052), as was the proportion of hospitalised patients (54.2% versus 38.9%, p = 0.0031). The mean number of anaesthesia exposures per individual was not statistically different (N = 0.958 ± 0.183/case versus 0.569 ± 0.107/control, p = 0.0732), but the proportion of children anaesthetised at least once was (43.8% (cases) versus 33.3% (controls), p = 0.0301). One or more hospitalisation(s) may reflect a health status and/or have an iatrogenic effect disrupting the normal setting up of learning abilities. Anaesthesia may play a role, but a correlation between LD and anaesthesia is of a lower magnitude than between LD and hospitalisation.


Mispredicting affective and behavioral responses to racism.
Contemporary race relations are marked by an apparent paradox: Overt prejudice is strongly condemned, yet acts of blatant racism still frequently occur. We propose that one reason for this inconsistency is that people misunderstand how they would feel and behave after witnessing racism. The present research demonstrates that although people predicted that they would be very upset by a racist act, when people actually experienced this event they showed relatively little emotional distress. Furthermore, people overestimated the degree to which a racist comment would provoke social rejection of the racist. These findings suggest that racism may persevere in part because people who anticipate feeling upset and believe that they will take action may actually respond with indifference when faced with an act of racism.


Involvement of P2X7 receptors in the regulation of neurotransmitter release in the rat hippocampus.
Although originally cloned from rat brain, the P2X7 receptor has only recently been localized in neurones, and functional responses mediated by these neuronal P2X7 receptors (P2X7 R) are largely unknown. Here we studied the effect of P2X7 R activation on the release of neurotransmitters from superfused rat hippocampal slices. ATP (1-30 mm) and other ATP analogues elicited concentration-dependent [3 H]GABA outflow, with the following rank order of potency: benzoylbenzoylATP (BzATP) > ATP > ADP. PPADS, the non-selective P2-receptor antagonist (3-30 microm), Brilliant blue G (1-100 nm) the P2X7 -selective antagonist and Zn2+ (0.1-30 microm) inhibited, whereas lack of Mg2+ potentiated the response by ATP. In situ hybridization revealed that P2X7 R mRNA is expressed in the neurones of the cell body layers in the hippocampus. P2X7 R immunoreactivity was found in excitatory synaptic terminals in CA1 and CA3 region targeting the dendrites of pyramidal cells and parvalbumin labelled structures. ATP (3-30 microm) and BzATP (0.6-6 microm) elicited concentration-dependent [14 C]glutamate efflux, and blockade of the kainate receptor-mediated transmission by CNQX (10-100 microm) and gadolinium (100 microm), decreased ATP evoked [3 H]GABA efflux. The Na+ channel blocker TTX (1 microm), low temperature (12 degrees C), and the GABA uptake blocker nipecotic acid (1 mm) prevented ATP-induced [3 H]GABA efflux. Brilliant blue G and PPADS also reduced electrical field stimulation-induced [3 H]GABA efflux. In conclusion, P2X7 Rs are localized to the excitatory terminals in the hippocampus, and their activation regulates the release of glutamate and GABA from themselves and from their target cells.


Can human nutrition be improved through better fish feeding practices? a review paper.
Achieving Sustainable Development Goal 2 of zero hunger and malnutrition by 2030 will require dietary shifts that include increasing the consumption of nutrient dense foods by populations in low- and middle-income countries. Animal source foods are known to be rich in a number of highly bioavailable nutrients that otherwise are not often consumed in the staple-food based diets of poorer populations throughout the world. Fish is the dominant animal source food in many low- and middle-income countries in the global south and is available from both fisheries and aquaculture. Consumers often perceive that wild caught fish have higher nutritional value than fish produced through aquaculture, and this may be true for some nutrients, for example omega-3 fatty acid content. However, there is potential to modify the nutritional value of farmed fish through feeds and through production systems, illustrated by the common practice of supplementing omega-3 fatty acids in fish diets to optimize their fatty acid profile. This manuscript reviews the evidence related to fish feeds and the nutritional composition of fish with respect to a number of nutrients of interest to human health, including iron, zinc, vitamins A and D, selenium, calcium, and omega-3 fatty acids, with low- and middle-income country populations in mind. In general, we find that the research on fortification of fish diet particularly with vitamins and minerals has not been directed toward human health but rather toward improvement of fish growth and health performance. We were unable to identify any studies directly exploring the impact of fish feed modification on the health of human consumers of fish, but as nutrition and health rises in the development agenda and consumer attention, the topic requires more urgent attention in future feed formulations.


Stability of external fixators used for knee arthrodesis after failed knee arthroplasty.
Secondary arthrodesis as treatment for failed knee arthroplasty is being used increasingly. The usual method is sustained rigid external fixation. The fusion rate is low in cases with considerable metaphyseal bone loss and poor bone quality. Currently, the Hoffmann- Vidal fixator or comparable designs are preferred to the less stable Charnley single frame fixator. The standard Hoffmann- Vidal device uses only transverse percutaneous fixating pins and gives an anteroposterior bending stiffness that is considerably lower than the lateral bending stiffness. In the present study, external fixators were tested for stability in a material testing machine under standardized conditions using synthetic bones. The anteroposterior bending stiffness was significantly improved when sagittal pins connected to a ventral compression rod were added to a Hoffmann- Vidal fixator. Stiffness was still more improved when sagittal pins were connected to ventral frames as in a modification of the Hoffmann- Vidal or the similar Ace-Fischer fixator.


Preparation and characterization of porous crosslinked collagenous matrices containing bioavailable chondroitin sulphate.
Porous collagen matrices with defined physical, chemical and biological characteristics are interesting materials for tissue engineering. Attachment of glycosaminoglycans (GAGs) may add to these characteristics and valorize collagen. In this study, porous type I collagen matrices were crosslinked using dehydrothermal (DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC), in the presence and absence of chondroitin sulphate (CS). EDC covalently attaches CS to collagen. DHT crosslinking preserved a porous matrix structure. However, attachment of CS to DHT-treated matrices using EDC, resulted in collapsed surfaces, CS located only at the matrix exterior. EDC crosslinking resulted in a partial matrix collapse. This could be prevented if crosslinking was carried out in the presence of ethanol. Matrix porosity was then preserved. The presence of CS during EDC crosslinking resulted in covalent immobilization of CS throughout the matrix. The amount of CS incorporated was increased if crosslinking was performed in the presence of ethanol. EDC-crosslinked matrices, with and without CS, had increased denaturation temperatures and decreased free amine group contents. The susceptibility of these matrices towards degradation by proteolytic enzymes was diminished. Immobilized CS increased the water-binding capacity and decreased the denaturation temperature and tensile strength. Immobilized CS bound anti-CS antibodies and was susceptible to chondroitinase ABC digestion, demonstrating its bioavailability. The modified matrices were not cytotoxic as was established using human myoblast and fibroblast culture systems. It is concluded that the use of ethanol during EDC crosslinking, offers an elegant means for the preparation of defined porous collagenous matrices containing bioavailable, covalently attached CS.


Bradyrhizobium japonicum has two differentially regulated, functional homologs of the sigma 54 gene (rpoN).
Recognition of -24/-12-type promoters by RNA polymerase requires a special sigma factor, sigma 54 (RpoN NtrA GlnF). In the nitrogen-fixing soybean symbiont Bradyrhizobium japonicum, two functional, highly conserved rpoN genes (rpoN1 and rpoN2) were identified and sequenced. The two predicted B. japonicum RpoN protein sequences were 87% identical, and both showed different levels of homology to the RpoN proteins of other bacteria. Downstream of rpoN2 (but not of rpoN1), two additional open reading frames were identified that corresponded to open reading frames located at similar positions in Klebsiella pneumoniae and Pseudomonas putida. Both B. japonicum rpoN genes complemented the succinate- and nitrate-negative phenotypes of a Rhizobium meliloti rpoN mutant. B. japonicum strains carrying single or double rpoN mutations were still able to utilize C4-dicarboxylates as a carbon source and histidine, proline, or arginine as a nitrogen source, whereas the ability to assimilate nitrate required expression of at least one of the two rpN genes. In symbiosis both rpoN genes could replace each other functionally. The rpoN1/2 double mutant induced about twice as many nodules on soybeans as did the wild type, and these nodules lacked nitrogen fixation activity completely. Transcription of a nifH'-'lacZ fusion was not activated in the rpoN1/2 mutant background, whereas expression of a fixR'-'lacZ fusion in this mutant was affected only marginally. By using rpoN'-'lacZ fusions, rpoN1 expression was shown to be activated at least sevenfold in microaerobiosis as compared with that in aerobiosis, and this type of regulation involved fixLJ. Expression of rpoN2 was observed under all conditions tested and was increased fivefold in an rpoN2 mutant. The data suggested that the rpoN1 gene was regulated in response to oxygen, whereas the rpoN2 gene was negatively autoregulated.


Saccade-target selection of dyslexic children when reading Chinese.
This study investigates the eye movements of dyslexic children and their age-matched controls when reading Chinese. Dyslexic children exhibited more and longer fixations than age-matched control children, and an increase of word length resulted in a greater increase in the number of fixations and gaze durations for the dyslexic than for the control readers. The report focuses on the finding that there was a significant difference between the two groups in the fixation landing position as a function of word length in single-fixation cases, while there was no such difference in the initial fixation of multi-fixation cases. We also found that both groups had longer incoming saccade amplitudes while the launch sites were closer to the word in single fixation cases than in multi-fixation cases. Our results suggest that dyslexic children's inefficient lexical processing, in combination with the absence of orthographic word boundaries in Chinese, leads them to select saccade targets at the beginning of words conservatively. These findings provide further evidence for parafoveal word segmentation during reading of Chinese sentences.


Transition planning for students with intellectual disability, autism, or other disabilities: data from the National Longitudinal Transition Study-2.
To compare the status of transition planning for students with intellectual disability, autism, or other disabilities, we used data from the National Longitudinal Transition Study-2, a federally funded, national study of the secondary and postschool experiences of students with disabilities. Results show that although transition planning had been conducted for the majority of students, few of them took a leadership role in their transition planning. Students with autism or intellectual disability were significantly less likely than students with other disabilities to take a leadership role. The majority of the active participants in transition planning were school-based personnel. We also found limited participation from other agencies/support persons (e.g., vocational rehabilitation). Students with autism or intellectual disability had more identified needs for support after school than did students with other disabilities.


New insights on taxonomy, phylogeny and population genetics of Leishmania (Viannia) parasites based on multilocus sequence analysis.
The Leishmania genus comprises up to 35 species, some with status still under discussion. The multilocus sequence typing (MLST)--extensively used for bacteria--has been proposed for pathogenic trypanosomatids. For Leishmania, however, a detailed analysis and revision on the taxonomy is still required. We have partially sequenced four housekeeping genes--glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), mannose phosphate isomerase (MPI) and isocitrate dehydrogenase (ICD)--from 96 Leishmania (Viannia) strains and assessed their discriminatory typing capacity. The fragments had different degrees of diversity, and are thus suitable to be used in combination for intra- and inter-specific inferences. Species-specific single nucleotide polymorphisms were detected, but not for all species; ambiguous sites indicating heterozygosis were observed, as well as the putative homozygous donor. A large number of haplotypes were detected for each marker; for 6PGD a possible ancestral allele for L. (Viannia) was found. Maximum parsimony-based haplotype networks were built. Strains of different species, as identified by multilocus enzyme electrophoresis (MLEE), formed separated clusters in each network, with exceptions. NeighborNet of concatenated sequences confirmed species-specific clusters, suggesting recombination occurring in L. braziliensis and L. guyanensis. Phylogenetic analysis indicates L. lainsoni and L. naiffi as the most divergent species and does not support L. shawi as a distinct species, placing it in the L. guyanensis cluster. BURST analysis resulted in six clonal complexes (CC), corresponding to distinct species. The L. braziliensis strains evaluated correspond to one widely geographically distributed CC and another restricted to one endemic area. This study demonstrates the value of systematic multilocus sequence analysis (MLSA) for determining intra- and inter-species relationships and presents an approach to validate the species status of some entities. Furthermore, it contributes to the phylogeny of L. (Viannia) and might be helpful for epidemiological and population genetics analysis based on haplotype/diplotype determinations and inferences.


Safety, tolerability, and efficacy of high-frequency chest wall oscillation in pediatric patients with cerebral palsy and neuromuscular diseases: an exploratory randomized controlled trial.
Airway secretions and infections are common in cerebral palsy and neuromuscular diseases. Chest physiotherapy is standard therapy but effort is substantial. High-frequency chest wall oscillation is used in cystic fibrosis but tolerability and safety data in cerebral palsy and neuromuscular disease are limited. A prospective, randomized, controlled trial of high-frequency chest wall oscillation and standard chest physiotherapy was performed in participants with neuromuscular disease or cerebral palsy. Outcome measures included respiratory-related hospitalizations, antibiotic therapy, chest radiographs, and polysomnography. Care-givers were questioned regarding therapy adherence. A total of 28 participants enrolled, 23 completed (12 chest physiotherapy, mean study period 5 months). No adverse outcomes were reported. Adherence to prescribed regimen was higher with high-frequency chest wall oscillation (P = .036). Our data suggest safety, tolerability, and better compliance with high-frequency chest wall oscillation. Improvement in airway clearance may help prevent hospitalizations. Larger controlled trials are required to confirm these results.


Gonadal hormone modulation of ∆9-tetrahydrocannabinol-induced antinociception and metabolism in female versus male rats.
The gonadal hormones testosterone (T) in adult males and estradiol (E2) in adult females have been reported to modulate behavioral effects of ∆9-tetrahydrocannabinol (THC). This study determined whether activational effects of T and E2 are sex-specific, and whether hormones modulate production of the active metabolite 11-hydroxy-THC (11-OH-THC) and the inactive metabolite 11-nor-9-carboxy-THC (THC-COOH). Adult male and female rats were gonadectomized (GDX) and treated with nothing (0), T (10-mm Silastic capsule/100g body weight), or E2 (1-mm Silastic capsule/rat). Three weeks later, saline or the cytochrome P450 inhibitor proadifen (25mg/kg; to block THC metabolism and boost THC's effects) was injected i.p.; 1h later, vehicle or THC (3mg/kg females, 5mg/kg males) was injected i.p., and rats were tested for antinociceptive and motoric effects 15-240min post-injection. T did not consistently alter THC-induced antinociception in males, but decreased it in females (tail withdrawal test). Conversely, T decreased THC-induced catalepsy in males, but had no effect in females. E2 did not alter THC-induced antinociception in females, but enhanced it in males. The discrepant effects of T and E2 on males' and females' behavioral responses to THC suggests that sexual differentiation of THC sensitivity is not simply due to activational effects of hormones, but also occurs via organizational hormone or sex chromosome effects. Analysis of serum showed that proadifen increased THC levels, E2 increased 11-OH-THC in GDX males, and T decreased 11-OH-THC (and to a lesser extent, THC) in GDX females. Thus, hormone modulation of THC's behavioral effects is caused in part by hormone modulation of THC oxidation to its active metabolite. However, the fact that hormone modulation of metabolism did not alter THC sensitivity similarly on all behavioral measures within each sex suggests that other mechanisms also play a role in gonadal hormone modulation of THC sensitivity in adult rats.


Greater responsiveness to donepezil in Alzheimer patients with higher levels of acetylcholinesterase based on attention task scores and a donepezil PET study.
The aim of the study was to predict donepezil responders among patients with Alzheimer disease (AD) based on cognitive tests and positron emission tomography. The Mini-Mental State Examination, Digit Symbol subtest (DigSm) of Wechsler Adult Intelligence Scale Revised, and Trail-Making Test A were administered for 80 patients with AD to assess global function, attention, and executive function, respectively. The same tests and the Clinical Global Impression (CGI) scale were conducted after treatment with oral donepezil (5 mg/d) for 6 months (study 1). [C]-Donepezil positron emission tomography examinations were conducted before and after treatment for 30 randomly selected patients. The distribution volume (DV), which indicates the density of donepezil-binding sites, was calculated using Logan graphical analysis (study 2). In study 1, 35 patients were identified as responders based on the CGI and Mini-Mental State Examination changes. These patients had higher baseline DigSm scores compared with nonresponders. In study 2, 15 patients were responders. DigSm correlated with DV at baseline. DV at baseline and %DV change in responders were higher than in nonresponders, and these variables correlated with ΔDigSm and CGI scores. Higher baseline attention may predict responsiveness to donepezil in patients with AD, and higher acetylcholinesterase levels result in a greater clinical effect.


Decongestants and antihistamines for acute otitis media in children.
Acute otitis media (AOM) is a common and important source of morbidity in children, although the majority of cases resolve spontaneously. While frequently recommended, decongestant and antihistamine therapy is of unclear benefit. To determine the efficacy of decongestant and antihistamine therapy in children with AOM on outcomes of AOM resolution, symptom resolution, medication side effects, and complications of AOM. In this updated review, we searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007, Issue 2); MEDLINE (January 2004 to May 2007); and EMBASE (July 2003 to May 2007). Randomized controlled trials (RCTs) evaluating decongestant or antihistamine treatment for children with AOM were included. Patient-oriented outcomes were considered most relevant. The review authors independently evaluated studies for inclusion, performed validity assessments and completed data extraction. Dichotomous data were pooled to generate relative risks; homogeneity was assessed using approximate chi-square tests. No new studies were included following this updated search. Fifteen trials involving 2695 people were included. Only the combined decongestant-antihistamine group demonstrated statistically lower rates of persistent AOM at the two week period (fixed relative risk (RR) 0.76; 95% confidence interval (CI) 0.60 to 0.96; number needed to treat (NNT) 10). No benefit was found for early cure rates, symptom resolution, prevention of surgery or other complications. There was a five to eight -fold increased risk of side effects for those receiving an intervention, which reached statistical significance for all decongestant groupings. Validity sub analyses demonstrated that lower quality studies found benefit, while analysis of those studies with higher validity scores found no benefit of treatment. Given lack of benefit and increased risk of side effects, these data do not support the use of decongestant treatment in children with AOM. There was a small statistical benefit from combination medication use but the clinical significance is minimal and study design may be biasing the results. Thus, the routine use of antihistamines for treating AOM in children cannot be recommended.


[Ecthyma gangrenosum associated with infection involving a methicillin-sensitive, Panton-Valentine-negative strain of Staphylococcus aureus].
Ecthyma gangrenosum (EG) is an anatomoclinical syndrome commonly associated with Pseudomonas aeruginosa cutaneous infection. Other microorganisms have also been incriminated on occasion, with other viral, fungal and bacterial agents potentially causing EG. In this report, we present an extremely rare case of an EG caused by methicillin-sensitive Staphylococcus aureus (MSSA) infection. This case, highly characteristic of EG both clinically and histologically, calls into question the physiopathological mechanisms of the disease and provides a reminder that it may be caused by a variety of organisms. A 62-year-old woman, followed for HIV seropositivity at the AIDS stage, developed a painful purpuric skin rash evolving towards necrotic nodules characteristic of ecthyma gangrenosum. Skin biopsy confirmed the diagnosis of EG due to methicillin-sensitive S. aureus (MSSA) infection without toxins or bacteraemia. To the best of our knowledge, this is the first case in the literature in which MSSA is reported as the underlying cause of such lesions.


Temperature Spatially Offset Raman Spectroscopy (T-SORS): Subsurface Chemically Specific Measurement of Temperature in Turbid Media Using Anti-Stokes Spatially Offset Raman Spectroscopy.
Here we propose and demonstrate a new analytical method for the noninvasive measurement of subsurface temperatures within diffusely scattering (turbid) media in combination with high chemical selectivity. The method is based upon the first combination of Stokes/anti-Stokes light scattering measurements and the recently developed spatially offset Raman spectroscopy (SORS). This approach has been conceptually demonstrated by measuring material-specific temperatures within a turbid sublayer of poly(tetrafluoroethylene) (PTFE) through a highly diffusely scattering overlayer of poly(oxymethylene) POM (3 mm thick). Root-mean-square errors (RMSEs) of 0.16-0.71 °C were achieved when measuring temperatures over ranges between 24 and 45 °C. This unique capability complements the array of existing, predominantly surface-based, temperature measurement techniques. It paves the way for a wide range of topical applications including subsurface, chemically specific, noninvasive temperature measurements within translucent media including the human body, subsurface monitoring of chemical or catalytic processes in manufacture quality and process control, and research.


Computational redesign of the SHV-1 beta-lactamase/beta-lactamase inhibitor protein interface.
Beta-lactamases are enzymes that catalyze the hydrolysis of beta-lactam antibiotics. beta-lactamase/beta-lactamase inhibitor protein (BLIP) complexes are emerging as a well characterized experimental model system for studying protein-protein interactions. BLIP is a 165 amino acid protein that inhibits several class A beta-lactamases with a wide range of affinities: picomolar affinity for K1; nanomolar affinity for TEM-1, SME-1, and BlaI; but only micromolar affinity for SHV-1 beta-lactamase. The large differences in affinity coupled with the availability of extensive mutagenesis data and high-resolution crystal structures for the TEM-1/BLIP and SHV-1/BLIP complexes make them attractive systems for the further development of computational design methodology. We used EGAD, a physics-based computational design program, to redesign BLIP in an attempt to increase affinity for SHV-1. Characterization of several of designs and point mutants revealed that in all cases, the mutations stabilize the interface by 10- to 1000-fold relative to wild type BLIP. The calculated changes in binding affinity for the mutants were within a mean absolute error of 0.87 kcal/mol from the experimental values, and comparison of the calculated and experimental values for a set of 30 SHV-1/BLIP complexes yielded a correlation coefficient of 0.77. Structures of the two complexes with the highest affinity, SHV-1/BLIP (E73M) and SHV-1/BLIP (E73M, S130K, S146M), are presented at 1.7 A resolution. While the predicted structures have much in common with the experimentally determined structures, they do not coincide perfectly; in particular a salt bridge between SHV-1 D104 and BLIP K74 is observed in the experimental structures, but not in the predicted design conformations. This discrepancy highlights the difficulty of modeling salt bridge interactions with a protein design algorithm that approximates side chains as discrete rotamers. Nevertheless, while local structural features of the interface were sometimes miscalculated, EGAD is globally successful in designing complexes with increased affinity.


Maternal Telomere Length and Risk of Down Syndrome: Epidemiological Impact of Smokeless Chewing Tobacco and Oral Contraceptive on Segregation of Chromosome 21.
We have previously demonstrated a relationship between children born with Down syndrome and maternal telomere length. Similarly, exposure to tobacco and oral contraceptives has been explored in one of our earlier studies as a risk factor for Down syndrome. In the present study, we consider the interactions among these risk factors associated with Down syndrome in a population from Kolkata, India, using analyses stratified by maternal age. We estimated the telomere length of women with children with Down syndrome by restriction enzyme/Southern blot methods. Linear regression was employed to estimate telomere shortening as an indicator of the maternal age of conception. Interactions among the various factors were analyzed by logistic regression. We found an association between the use of smokeless chewing tobacco and shorter telomere length among women who experienced meiosis I nondisjunction at gametogenesis; the effect is seen across all maternal age groups. In contrast, oral contraceptive use alone did not exhibit a statistically significant association with maternal telomere length, but there was an interaction with the use of smokeless chewing tobacco in the older mothers who experienced meiotic II nondisjunction. Environmental/habitual factors interact with molecular components of the oocyte, which ultimately increases the risk of chromosome 21 nondisjunction and subsequently of giving birth to a child with Down syndrome.


Causes and risk factors for death in systemic sclerosis: a study from the EULAR Scleroderma Trials and Research (EUSTAR) database.
To determine the causes and predictors of mortality in systemic sclerosis (SSc). Patients with SSc (n=5860) fulfilling the American College of Rheumatology criteria and prospectively followed in the EULAR Scleroderma Trials and Research (EUSTAR) cohort were analysed. EUSTAR centres completed a structured questionnaire on cause of death and comorbidities. Kaplan-Meier and Cox proportional hazards models were used to analyse survival in SSc subgroups and to identify predictors of mortality. Questionnaires were obtained on 234 of 284 fatalities. 55% of deaths were attributed directly to SSc and 41% to non-SSc causes; in 4% the cause of death was not assigned. Of the SSc-related deaths, 35% were attributed to pulmonary fibrosis, 26% to pulmonary arterial hypertension (PAH) and 26% to cardiac causes (mainly heart failure and arrhythmias). Among the non-SSc-related causes, infections (33%) and malignancies (31%) were followed by cardiovascular causes (29%). Of the non-SSc-related fatalities, 25% died of causes in which SSc-related complications may have participated (pneumonia, sepsis and gastrointestinal haemorrhage). Independent risk factors for mortality and their HR were: proteinuria (HR 3.34), the presence of PAH based on echocardiography (HR 2.02), pulmonary restriction (forced vital capacity below 80% of normal, HR 1.64), dyspnoea above New York Heart Association class II (HR 1.61), diffusing capacity of the lung (HR 1.20 per 10% decrease), patient age at onset of Raynaud's phenomenon (HR 1.30 per 10 years) and the modified Rodnan skin score (HR 1.20 per 10 score points). Disease-related causes, in particular pulmonary fibrosis, PAH and cardiac causes, accounted for the majority of deaths in SSc.


A Novel Effector Gene SCRE2 Contributes to Full Virulence of Ustilaginoidea virens to Rice.
Ustilaginoidea virens, the causal agent of rice false smut (RFS), has become one of the most devastating rice pathogens worldwide. As a group of essential virulence factors, the effectors in the filamentous fungus might play central roles in the interaction between plants and pathogens. However, little is known about the roles of individual effectors in U. virens virulence. In this study, we identified and characterized a small secreted cysteine-rich effector, SCRE2, in U. virens. SCRE2 was first confirmed as an effector through yeast secretion, protein localization and translocation assays, as well as its expression pattern during U. virens infection. Transient expression of SCRE2 in Nicotiana benthamiana suppressed necrosis-like defense symptoms triggered by the mammalian BAX and oomycete elicitin INF1 proteins. The ability of SCRE2 to inhibit immunity-associated responses in N. benthamiana, including elicitor-triggered cell death and oxidative burst, is further defined to a small peptide region SCRE268-85 through expressing a series of truncated proteins. Convincingly, ectopic expression of SCRE2 in the transgenic rice cells significantly inhibited pathogen-associated molecular pattern-triggered immunity including flg22- and chitin-induced defense gene expression and oxidative burst. Furthermore, the scre2 knockout mutant generated by the CRISPR/Cas9 system greatly attenuated in U. virens virulence to rice. Collectively, this study indicates that the effector SCRE2 is able to inhibit plant immunity and is required for full virulence of U. virens.