NFκB is a central regulator of protein quality control in response to protein aggregation stresses via autophagy modulation.
During cell life, proteins often misfold, depending on particular mutations or environmental changes, which may lead to protein aggregates that are toxic for the cell. Such protein aggregates are the root cause of numerous diseases called "protein conformational diseases," such as myofibrillar myopathy and familial amyotrophic lateral sclerosis. To fight against aggregates, cells are equipped with protein quality control mechanisms. Here we report that NFκB transcription factor is activated by misincorporation of amino acid analogues into proteins, inhibition of proteasomal activity, expression of the R120G mutated form of HspB5 (associated with myofibrillar myopathy), or expression of the G985R and G93A mutated forms of superoxide dismutase 1 (linked to familial amyotrophic lateral sclerosis). This noncanonical stimulation of NFκB triggers the up-regulation of BAG3 and HspB8 expression, two activators of selective autophagy, which relocalize to protein aggregates. Then NFκB-dependent autophagy allows the clearance of protein aggregates. Thus NFκB appears as a central and major regulator of protein aggregate clearance by modulating autophagic activity. In this context, the pharmacological stimulation of this quality control pathway might represent a valuable strategy for therapies against protein conformational diseases.


Elevated serum osteopontin levels and genetic polymorphisms of osteopontin are associated with Vogt-Koyanagi-Harada disease.
Osteopontin (OPN) is a proinflammatory cytokine involved in chronic inflammatory diseases. This study aimed to analyze the role of OPN in the pathogenesis of Vogt-Koyanagi-Harada (VKH) disease. Serum levels of OPN in VKH patients and healthy controls were assayed by enzyme-linked immunosorbent assay (ELISA). Peripheral blood mononuclear cells (PBMCs) or CD4+ T cells were cultured with anti-CD3 and anti-CD28 antibodies in the absence or presence of recombinant OPN for the determination of cell proliferation and cytokines. Cell proliferation was detected using a cell counting kit. Levels of interferon (IFN)-γ and interleukin (IL)-17 were detected by ELISA. Four single nucleotide polymorphisms (SNPs) of OPN and four SNPs of OPN receptors were genotyped in 601 VKH patients and 605 healthy controls using a polymerase chain reaction-restriction fragment length polymorphism assay. OPN serum levels were significantly higher in patients with active VKH than in patients with inactive VKH and in healthy controls. PBMCs or CD4+ T cells cultured with recombinant OPN induced a marked cell proliferation and profound secretion of IFN-γ and IL-17 from patients with active VKH. A significantly increased frequency of the OPN rs4754 TT genotype (P = 0.004, pc = 0.048) was observed in VKH patients compared with healthy controls. No association could be detected among the four selected SNPs of OPN receptors and VKH. OPN may be relevant to the pathogenesis of VKH disease. The TT genotype of rs4754 may be a susceptible factor for VKH disease in a Chinese Han population.


Tractional retinal detachment following intravitreal bevacizumab (Avastin) in patients with severe proliferative diabetic retinopathy.
The aim of this study was to report the development or progression of tractional retinal detachment (TRD) after the injection of intravitreal bevacizumab (Avastin) used as an adjuvant to vitrectomy for the management of severe proliferative diabetic retinopathy (PDR). The clinical charts of patients who experienced the development or progression of TRD after an intravitreal injection of 1.25 mg bevacizumab before vitrectomy for the management of PDR were reviewed. Eleven eyes (patients) out of 211 intravitreal injections (5.2%) that developed or had progression of TRD were identified. All eyes had PDR refractory to panretinal photocoagulation (PRP). Nine patients had type 1 diabetes mellitus (DM), and two patients had type 2 DM. Patients had a mean age of 39.5 years (range 22-62 years). In the current study, all patients used insulin administration and had poor glycaemic control (mean HbA(1c) 10.6%). Time from injection to TRD was a mean of 13 days (range 3-31 days). Mean best correct visual acuity (BCVA) at TRD development or progression was logarithm of the minimal angle of resolution (LogMAR) 2.2 (range 1.0-2.6) (mean Snellen equivalent hand motions; range 20/200 to light perception), a statistically significant worsening compared with baseline BCVA (p<0.0001). Eight eyes underwent vitrectomy and three patients refused or were unable to undergo surgery. The final mean BCVA after surgery was LogMAR 0.9 (range 0.2-2.0) (mean Snellen equivalent 20/160; range 20/32 to counting fingers), a statistically significant improvement compared with TRD BCVA (p = 0.002). TRD may occur or progress shortly following administration of intravitreal bevacizumab in patients with severe PDR.


Prevalence and Correlates of CKD in Hispanics/Latinos in the United States.
The prevalence of ESRD among Hispanics/Latinos is 2-fold higher than in non-Hispanic whites. However, little is known about the prevalence of earlier stages of CKD among Hispanics/Latinos. This study estimated the prevalence of CKD in US Hispanics/Latinos. This was a cross-sectional study of 15,161 US Hispanic/Latino adults of Cuban, Dominican, Mexican, Puerto Rican, Central American, and South American backgrounds enrolled in the multicenter, prospective, population-based Hispanic Community Health Study/Study of Latinos (HCHS/SOL). In addition, the prevalence of CKD in Hispanics/Latinos was compared with other racial/ethnic groups in the 2007-2010 National Health and Nutrition Examination Survey (NHANES). Prevalent CKD was defined as an eGFR <60 ml/min per 1.73 m(2) (estimated with the 2012 Chronic Kidney Disease Epidemiology Collaboration eGFR creatinine-cystatin C equation) or albuminuria based on sex-specific cut points determined at a single point in time. The overall prevalence of CKD among Hispanics/Latinos was 13.7%. Among women, the prevalence of CKD was 13.0%, and it was lowest in persons with South American background (7.4%) and highest (16.6%) in persons with Puerto Rican background. In men, the prevalence of CKD was 15.3%, and it was lowest (11.2%) in persons with South American background and highest in those who identified their Hispanic background as "other" (16.0%). The overall prevalence of CKD was similar in HCHS/SOL compared with non-Hispanic whites in NHANES. However, prevalence was higher in HCHS/SOL men and lower in HCHS/SOL women versus NHANES non-Hispanic whites. Low income, diabetes mellitus, hypertension, and cardiovascular disease were each significantly associated with higher risk of CKD. Among US Hispanic/Latino adults, there was significant variation in CKD prevalence among Hispanic/Latino background groups, and CKD was associated with established cardiovascular risk factors.


Stressful life events and depressive symptoms in young adolescents: Modulation by respiratory sinus arrhythmia? The TRAILS study.
Respiratory sinus arrhythmia (RSA) has been proposed as a physiological marker of emotion-regulation capacity, and shown to be cross-sectionally associated with depression. Little is known about the role of RSA as a predictor of (subclinical) depressive symptoms over time and as a modifier of the depressogenic effect of stressful life events (SLEs). In a longitudinal population-based study with data collected in 1653 adolescents twice (at age 11 and 13.5 years, respectively), RSA was assessed in supine position at the first assessment wave. Depressive symptoms were assessed at both waves and SLEs experienced between the two waves at the last wave. Low levels of RSA were not associated with concurrent or future depressive symptoms, and did not enhance the depressogenic effects of SLEs. In a normal population of young adolescents, a low level of RSA does not identify adolescents at risk for depressive symptoms when confronted with SLEs. In post hoc analyses, among those reporting high exposure to stressful life events, higher RSA tended to predict less self-reported anxiety and more self-reported somatic symptoms as compared to those with lower RSA.


Pulsed electrospray for mass spectrometry.
Pulsed electrospray has been developed and been reported for the first time. This new technique is based on the principle of pulsed ion sources, combined with the conventional electrospray. The pulsed ion spray was realized by a homemade pulsed HV circuit and was monitored by a digital microscope and an oscilloscope. Results show that the pulsed ESI device can be operated under proper conditions for a clear on-and-off spray process and that the device was kept in good electric contact for electrospray when pulsed HV was on. A pulsed ion current and pulsed mass spectra can be achieved with this pulsed ESI device. Furthermore, it has been noted that, under the same conditions (i.e., shape and size of sprayer tip, distance from sprayer tip to sampling nozzle, and other parameters for mass spectrometer), stable electrospray could be obtained for lower flow rates with a pulsed spray device. This experimental fact indicates the possible reduction in the total sample consumed could be realized by exploiting this novel design.


Signatures of natural selection between life cycle stages separated by metamorphosis in European eel.
Species showing complex life cycles provide excellent opportunities to study the genetic associations between life cycle stages, as selective pressures may differ before and after metamorphosis. The European eel presents a complex life cycle with two metamorphoses, a first metamorphosis from larvae into glass eels (juvenile stage) and a second metamorphosis into silver eels (adult stage). We tested the hypothesis that different genes and gene pathways will be under selection at different life stages when comparing the genetic associations between glass eels and silver eels. We used two sets of markers to test for selection: first, we genotyped individuals using a panel of 80 coding-gene single nucleotide polymorphisms (SNPs) developed in American eel; second, we investigated selection at the genome level using a total of 153,423 RAD-sequencing generated SNPs widely distributed across the genome. Using the RAD approach, outlier tests identified a total of 2413 (1.57%) potentially selected SNPs. Functional annotation analysis identified signal transduction pathways as the most over-represented group of genes, including MAPK/Erk signalling, calcium signalling and GnRH (gonadotropin-releasing hormone) signalling. Many of the over-represented pathways were related to growth, while others could result from the different conditions that eels inhabit during their life cycle. The observation of different genes and gene pathways under selection when comparing glass eels vs. silver eels supports the adaptive decoupling hypothesis for the benefits of metamorphosis. Partitioning the life cycle into discrete morphological phases may be overall beneficial since it allows the different life stages to respond independently to their unique selection pressures. This might translate into a more effective use of food and niche resources and/or performance of phase-specific tasks (e.g. feeding in the case of glass eels, migrating and reproducing in the case of silver eels).


The Influence of Propofol and Sevoflurane on Acute Kidney Injury After Colorectal Surgery: A Retrospective Cohort Study.
Acute kidney injury (AKI) is a primary cause of morbidity and mortality after major abdominal surgery; however, little is known about the effect of anesthetics on the development of AKI after colorectal surgery. The objective of this study was to compare the effects of anesthesia with propofol and sevoflurane on postoperative AKI after colorectal surgery. For this study, we reviewed the electronic medical records of 4320 patients who underwent colorectal surgeries between January 2008 and December 2011. The influence of propofol and sevoflurane on the development of postoperative AKI and short-term outcomes was assessed by multivariable analysis, and the effect of the anesthetic agent on overall mortality was analyzed by a Cox proportional hazard model with propensity score matching method. Overall, the incidence of AKI was 9.6% by Acute Kidney Injury Network (AKIN) criteria and 5.8% by risk, injury, failure, loss, and end-stage renal disease (RIFLE) criteria. The incidence of AKI incidence was greater in patients receiving sevoflurane than those receiving propofol (142 [11.2%] vs 272 [8.9%], P = 0.02 by AKIN criteria, 94 [7.4%] vs 157 [5.1%], P = 0.004 by RIFLE criteria). Multivariate logistic regression and propensity score matching results indicated that, when compared with propofol, sevoflurane anesthesia may be associated with the development of postoperative AKI (odds ratio [OR], 1.29; P = 0.03; and OR, 1.44; P = 0.02 by AKIN and RIFLE criteria, and OR, 1.41; P = 0.04 by RIFLE criteria, respectively). We found no relationship between sevoflurane and overall mortality. Compared with anesthesia with propofol, anesthesia with sevoflurane may be associated with a modest increase in the incidence of AKI when RIFLE but not AKIN criteria are used. Thus, the clinical meaning of these results is uncertain. Further work is needed to clarify the relevance of such an association.


The Reliability and Factorial Validity of the Swedish Version of the Revised Controlling Behaviors Scale.
This study focused on the factor structure of the victimization form of the revised Controlling Behaviors Scale (CBS-R). Data from 1,218 women and men were analyzed in the study. Results of confirmatory factor analysis (CFA) failed to find support for the proposed five-factor structure of the scale, as the items on the scale were better represented by one common factor. In addition, when examining if controlling behaviors are distinct from psychological aggression, the CFA indicated that the items on the CBS-R are clearly distinguishable from the items on the psychological aggression (as measured with the subscales of the revised Conflict Tactic Scales [CTS2]), and that this holds for both males and females. Implications for the general use of the CBS-R and for use in conjunction with psychological aggression and physical aggression in intimate partner violence were discussed.


Bendamustine in Combination With Gemcitabine and Vinorelbine Is an Effective Regimen As Induction Chemotherapy Before Autologous Stem-Cell Transplantation for Relapsed or Refractory Hodgkin Lymphoma: Final Results of a Multicenter Phase II Study.
This multicenter, open-label, phase II study evaluated the combination of bendamustine, gemcitabine, and vinorelbine (BeGEV) as induction therapy before autologous stem-cell transplantation (ASCT) in patients with relapsed or refractory Hodgkin lymphoma (HL). Patients with HL who were refractory to or had relapsed after one previous chemotherapy line were eligible. The primary end point was complete response (CR) rate after four cycles of therapy. Secondary end points were: overall response rate, stem-cell mobilization activity, and toxicity. Progression-free and overall survival were also evaluated. In total, 59 patients were enrolled. After four cycles of therapy, 43 patients (73%) achieved CR, and six (10%) achieved partial response, for an overall response rate of 83%. The most common grade 3 to 4 nonhematologic toxicities included febrile neutropenia (n = 7) and infection (n = 4). Regarding hematologic toxicities, grade 3 to 4 thrombocytopenia and neutropenia were each experienced by eight patients (13.5%). CD34+ cells were successfully harvested in 55 of 57 evaluable patients, and 43 of 49 responding patients underwent ASCT. With a median follow-up of 29 months, the 2-year progression-free and overall survival rates for the total population were 62.2% and 77.6%, respectively. The same figures for patients undergoing autograft were 80.8% and 89.3%, respectively. This phase II study demonstrates that BeGEV is an effective salvage regimen able to induce CR in a high proportion of patients with relapsed or refractory HL before ASCT. These data provide a strong rationale for further development of the BeGEV regimen.


Premedication with pronase or N-acetylcysteine improves visibility during gastroendoscopy: an endoscopist-blinded, prospective, randomized study.
To assess the efficacy of premedicaton with pronase or N-acetylcysteine (NAC) at 20 min before upper gastrointestinal (UGI) endoscopy and to determine whether pronase or NAC pretreatment influences the reliability of the rapid urease test. A total of 146 patients were prospectively and randomly assigned into the study groups according to different premedications before endoscopy. One endoscopist assessed mucosal visibility (MV) with scores ranged from 1 to 4 at four sites in the stomach. The sum of the MV scores from these four locations was defined as the total mucosal visibility (TMV) score. Identification of H pylori was performed using CLO test, histology, and serology. The Group with pronase premedication had a significantly lower TMV score than did the groups with gascon and gascon water (P < 0.001 and P < 0.01, respectively). The group with NAC had a significantly lower TMV score than the group with gascon (P < 0.01) and a trend of a lower MV score than the group with gascon water (P = 0.06). The TMV score did not significantly differ between the group with pronase and the group with NAC (P = 0.39 and P = 0.14, respectively). The sensitivity and specificity of the CLO test were 92.5% and 93.9%, respectively, in groups premedicated with pronase and NAC together. Premedication with pronase or NAC at 20 min before UGI endoscopy improves the mucosal visibility of the stomach. Neither pronase nor NAC produces any obvious interference with the CLO test for the identification of H pylori infection.


Circumcision and reduced risk of self-reported penile coital injuries: results from a randomized controlled trial in Kisumu, Kenya.
Injuries to the penis during intercourse represent a hypothesized mechanism by which uncircumcised men are at increased risk for HIV. There are no published, systematically collected data regarding mild penile coital trauma to our knowledge. We identified risks of self-reported penile coital injuries in men 18 to 24 years old in a randomized trial of circumcision to prevent HIV in Kisumu, Kenya. Each participant underwent standardized interview, medical history and physical examination at baseline, and 6, 12, 18 and 24 months after enrollment. Self-reported penile coital injuries were assessed at each visit, and were defined as penis feels sore during sex, penis gets scratches, cuts or abrasions during sex, and skin of the penis bleeds after sex. Generalized estimating equation analysis estimated odds ratios for penile coital injuries. From February 2002 to September 2005, 2,784 participants were randomized. At baseline 1,775 (64.4%) men reported any coital injury including 1,313 (47.6%) soreness, 1,328 (48.2%) scratches, abrasions or cuts and 461 (16.7%) bleeding. On multivariable analysis coital injury risk was lower for circumcised than for uncircumcised men with soreness (OR 0.71, 95% CI 0.64-0.80), scratches/abrasions/cuts (OR 0.52, 95% CI 0.46-0.59), bleeding (OR 0.62, 95% CI 0.51-0.75) and any coital injury (OR 0.61, 95% CI 0.54-0.68). Other significant risks included increasing age, multiple recent sex partners, HSV-2 seropositivity and genital ulcers (p <0.05). Condom use, cleaning the penis soon after intercourse and being married/cohabiting were protective (p <0.05, each). Self-reported penile coital injuries were common in these healthy young men. Circumcised men were at lower risk for coital injuries. Verifying penile coital injuries, the mechanism of acquisition and the association with HIV risk is needed.


Intracranial Aneurysms: Wall Motion Analysis for Prediction of Rupture.
Intracranial aneurysms are a common pathologic condition with a potential severe complication: rupture. Effective treatment options exist, neurosurgical clipping and endovascular techniques, but guidelines for treatment are unclear and focus mainly on patient age, aneurysm size, and localization. New criteria to define the risk of rupture are needed to refine these guidelines. One potential candidate is aneurysm wall motion, known to be associated with rupture but difficult to detect and quantify. We review what is known about the association between aneurysm wall motion and rupture, which structural changes may explain wall motion patterns, and available imaging techniques able to analyze wall motion.


Impedimetric thrombin aptasensor based on chemically modified graphenes.
Highly sensitive biosensors are of high importance to the biomedical field. Graphene represents a promising transducing platform for construction of biosensors. Here for the first time we compare the biosensing performance of a wide set of graphenes prepared by different methods. In this work, we present a simple and label-free electrochemical impedimetric aptasensor for thrombin based on chemically modified graphene (CMG) platforms such as graphite oxide (GPO), graphene oxide (GO), thermally reduced graphene oxide (TR-GO) and electrochemically reduced graphene oxide (ER-GO). Disposable screen-printed electrodes were first modified with chemically modified graphene (CMG) materials and used to immobilize a DNA aptamer which is specific to thrombin. The basis of detection relies on the changes in impedance spectra of redox probe after the binding of thrombin to the aptamer. It was discovered that graphene oxide (GO) is the most suitable material to be used as compared to the other three CMG materials. Furthermore, the optimum concentration of aptamer to be immobilized onto the modified electrode surface was determined to be 10 μM and the linear detection range of thrombin was 10-50 nM. Lastly, the aptasensor was found to demonstrate selectivity for thrombin. Such simply fabricated graphene oxide aptasensor shows high promise for clinical diagnosis of biomarkers and point-of-care analysis.


Insulin-like growth factor ligands, receptors, and binding proteins in cancer.
This review aims to summarize experimental evidence supporting the role of the insulin-like growth factor (IGF) signalling system in the progression, maintenance, and treatment of cancer. These data implicate the IGF system as an important modifier of cancer cell proliferation, survival, growth, and treatment sensitivity. The role of the IGF system in cancer should be examined in the context of the extra-cellular and intra-cellular signalling networks, in particular: phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt/PKB), mammalian target of rapamycin (mTOR), and forkhead transcription factors (FOXO). This review highlights evidence derived from molecular structure and functional genetics with respect to how the extra-cellular components of the IGF system function normally, and their subsequent modifications in cancer. The therapeutic relevance of the research evidence described is also addressed, as the challenge is to apply this knowledge to human health.


Lower Prevalence of Celiac Disease and Gluten-Related Disorders in Persons Living in Southern vs Northern Latitudes of the United States.
The association between prevalence of celiac disease and geographic region is incompletely understood, but the occurrence of several autoimmune disorders has been found to vary along a North-South gradient. We examined geographic, demographic, and clinical factors associated with prevalence of celiac disease and gluten-free diet in the United States. In a population-based study, we analyzed data on gluten-related conditions from the National Health and Nutrition Examination Survey, from 2009 through 2014, on 22,277 participants 6 years and older. We identified persons with celiac disease based on results of serum tests for IgA against tissue transglutaminase and endomysium or on both a health care provider diagnosis and adherence to a gluten-free diet. Gluten avoidance without celiac disease was defined as adherence to a gluten-free diet without a diagnosis of celiac disease. We compared mean serum levels of biochemical and nutritional markers based on status of gluten-related conditions. We found 0.7% of participants to have celiac disease and 1.1% of participants to avoid gluten without celiac disease. Celiac disease was more common among individuals who lived at latitudes of 35°-39° North (odds ratio, 3.2; 95% confidence interval, 1.4-7.1) or at latitudes of 40° North or more (odds ratio, 5.4; 95% CI, 2.6-11.3) than individuals who lived at latitudes below 35° North, independent of race or ethnicity, socioeconomic status, and body mass index. Gluten avoidance without celiac disease was more common among individuals who lived at latitudes of 40° North or more, independent of demographic factors and body mass index. Participants with undiagnosed celiac disease (identified by positive results from serologic tests) had lower mean levels of vitamin B-12 and folate (data collected from 2009 through 2012) than persons without celiac disease. Participants with a health care provider diagnosis of celiac disease had a lower mean level of hemoglobin than persons without celiac disease. Mean levels of albumin, calcium, iron, ferritin, cholesterol, vitamin B-6, and vitamin D (data collected from 2009 through 2010) did not differ between participants with gluten-related conditions and those without. In the US population, a higher proportion of persons living at latitudes of 35° North or greater have celiac disease or avoid gluten than persons living south of this latitude, independent of race or ethnicity, socioeconomic status, or body mass index. Mean levels of vitamin B-12 and folate are lower in individuals with undiagnosed celiac disease, and levels of hemoglobin are lower in participants with a diagnosis of celiac disease, compared with individuals without celiac disease.


Cryptosporidium parvum as a risk factor of diarrhea occurrence in neonatal alpacas in Peru.
Cryptosporidiosis has been reported as an important cause of neonatal diarrhea and mortality in cattle, sheep, and other ruminants, but its impact on alpaca health has not been studied thoroughly. In this study, we have determined the prevalence and evaluated the role of cryptosporidiosis as a risk factor for diarrhea occurrence in newborn alpacas. During the calving season (January-March) of 2006, stool specimens (N = 1312) were collected from 24 herds of newborn alpacas in Puno and Cuzco, departments that account for the largest populations of alpacas in Peru. All the specimens were microscopically screened for Cryptosporidium spp. using the acid-fast technique. The association between Cryptosporidium detection and diarrhea was analyzed using χ2 test and generalized lineal model. Cryptosporidium species were determined by PCR-RFLP analysis of the small subunit rRNA gene. Cryptosporidium oocysts were detected in 159 of 1312 (12.4%) newborn alpacas. Results of the analyses demonstrated that crypstosporidiosis was significantly associated with diarrhea (PR = 3.84; CI95% 2.54-5.81; p < 0.0001). Only Cryptosporidium parvum was detected in the 153 Cryptosporidium-infected animals. Thus, there is an association of C. parvum infection with diarrhea in neonatal alpacas.


[Implementation of a Preanesthetic Evaluation Service in a University Hospital: difficulties and results.].
Preoperative evaluation is the key for a good preoperative preparation and intraoperative handling. When done in the outpatient clinic, it allows for better analysis and further investigation, improving the quality of the preparation and decreasing hospital costs. The aim of this study was to analyze the implementation of the Serviço de Avaliação Pré-Anestésica (SAPAN Preanesthetic - Service) in a University Hospital, and to determine the difficulties and positive results. The data regarding patients seen at the SAPAN clinic, by the residents, under the supervision of an attending physician, in the first nine months after its implementation was reviewed. The consultation followed a standard form. At the end of the preanesthetic evaluation, patients were either cleared for surgery or referred to other specialties for the treatment of specific diseases. The items analyzed were: number of patients evaluated, number of elective surgeries performed during this period, surgical specialties involved, percentage of patients whose surgeries were cancelled due to the need to control underlying diseases, and clinical specialties requested the most to evaluate and treat those patients. Nine hundred and thirteen outpatient consultations and 5,409 elective surgeries were performed. In 11.9% of the cases, the surgeries were cancelled for medical reasons, and the most frequent specialties involved were cardiology (43.08%), pneumology (25.74%), hematology (21.65%), and endocrinology (9.52%). Although the majority of patients was not referred by the surgical specialties, the initial SAPAN data demonstrated the benefits of a preanesthetic evaluation service.


A hot spot for p53 mutation in transitional cell carcinoma of the bladder: clues to the etiology of bladder cancer.
Twenty-eight transitional cell carcinomas of the bladder, grade 2 or 3, were analyzed for the presence of p53 mutations. Thirteen tumors were found to contain 14 mutations. These were all base substitution mutations, of which nine were GC-->AT transitions (three at CpG sites). The remaining five mutations were transversions (three GC-->CG, one GC-->TA, and one AT-->TA). Four of the mutations were found at codon 280. A comparison with other studies of bladder tumors reveals that a region encompassing codons 280 and 285 represents a hot spot for p53 mutation in bladder cancer. The 280/285 hot spot lies within two purine-rich sequences that may provide some clues to the identity of potential bladder carcinogens. A comparison of mutations from bladder tumors of smokers and nonsmokers reveals no significant differences.


Placement of endovascular stented grafts via remote access sites: a new approach to the treatment of failed aortoiliofemoral reconstructions.
Endovascular grafting is a technique that combines the use of intravascular stents and prosthetic grafts to fabricate devices with unique properties. The purpose of this study is to describe the use of endovascular graft technology in the treatment of failed or failing standard aortoiliofemoral reconstructions. Over a 15-month period five patients with limb-threatening ischemia and failed aortofemoral or iliofemoral reconstructions underwent successful placement of six endovascular grafts to revascularize seven severely ischemic lower extremities. Standard thin-walled 6 mm polytetrafluoroethylene grafts and Palmaz balloon-expandable stents were used to fashion each reconstruction. In addition to the primary endovascular grafts, three patients underwent immediate femoropopliteal bypasses to improve distal outflow and one patient had a femorofemoral bypass graft to restore circulation to the contralateral ischemic extremity. The ankle/brachial indices of all patients significantly improved after the procedure (from a mean of 0.32 to a mean of 0.75) and all grafts are patent to date (mean follow-up 10 months). There were no deaths or limb loss in this group of patients. These favorable results indicate that this minimally invasive approach, which permits a new arterial graft to be inserted through a remote access site, is a valuable method for providing unobstructed arterial inflow after aortoiliofemoral graft failure. This procedure may be particularly valuable when there are contraindications to the use of axillary arteries or the thoracic aorta as alternatives to complex reoperative abdominal aortic surgery.


Isolation of singlet carbene derived 2-phospha-1,3-butadienes and their sequential one-electron oxidation to radical cations and dications.
A synthetic strategy for the 2-phospha-1,3-butadiene derivatives [{(IPr)C(Ph)}P(cAACMe)] (3a) and [{(IPr)C(Ph)}P(cAACCy)] (3b) (IPr = C{(NDipp)CH}2, Dipp = 2,6-iPr2C6H3; cAACMe = C{(NDipp)CMe2CH2CMe2}; cAACCy = C{(NDipp)CMe2CH2C(Cy)}, Cy = cyclohexyl) containing a C[double bond, length as m-dash]C-P[double bond, length as m-dash]C framework has been established. Compounds 3a and 3b have a remarkably small HOMO-LUMO energy gap (3a: 5.09; 3b: 5.05 eV) with a very high-lying HOMO (-4.95 eV for each). Consequently, 3a and 3b readily undergo one-electron oxidation with the mild oxidizing agent GaCl3 to afford radical cations [{(IPr)C(Ph)}P(cAACR)]GaCl4 (R = Me 4a, Cy 4b) as crystalline solids. The main UV-vis absorption band for 4a and 4b is red-shifted with respect to that of 3a and 3b, which is associated with the SOMO related transitions. The EPR spectra of compounds 4a and 4b each exhibit a doublet due to coupling of the unpaired electron with the 31P nucleus. Further one-electron removal from the radical cations 4a and 4b is also feasible with GaCl3, affording the dications [{(IPr)C(Ph)}P(cAACR)](GaCl4)2 (R = Me 5a, Cy 5b) as yellow crystals. The molecular structures of compounds 3-5 have been determined by X-ray diffraction and analyzed by DFT calculations.


Amino acid substitutions in the FXYD motif enhance phospholemman-induced modulation of cardiac L-type calcium channels.
We have found that phospholemman (PLM) associates with and modulates the gating of cardiac L-type calcium channels (Wang et al., Biophys J 98: 1149-1159, 2010). The short 17 amino acid extracellular NH(2)-terminal domain of PLM contains a highly conserved PFTYD sequence that defines it as a member of the FXYD family of ion transport regulators. Although we have learned a great deal about PLM-dependent changes in calcium channel gating, little is known regarding the molecular mechanisms underlying the observed changes. Therefore, we investigated the role of the PFTYD segment in the modulation of cardiac calcium channels by individually replacing Pro-8, Phe-9, Thr-10, Tyr-11, and Asp-12 with alanine (P8A, F9A, T10A, Y11A, D12A). In addition, Asp-12 was changed to lysine (D12K) and cysteine (D12C). As expected, wild-type PLM significantly slows channel activation and deactivation and enhances voltage-dependent inactivation (VDI). We were surprised to find that amino acid substitutions at Thr-10 and Asp-12 significantly enhanced the ability of PLM to modulate Ca(V)1.2 gating. T10A exhibited a twofold enhancement of PLM-induced slowing of activation, whereas D12K and D12C dramatically enhanced PLM-induced increase of VDI. The PLM-induced slowing of channel closing was abrogated by D12A and D12C, whereas D12K and T10A failed to impact this effect. These studies demonstrate that the PFXYD motif is not necessary for the association of PLM with Ca(V)1.2. Instead, since altering the chemical and/or physical properties of the PFXYD segment alters the relative magnitudes of opposing PLM-induced effects on Ca(V)1.2 channel gating, PLM appears to play an important role in fine tuning the gating kinetics of cardiac calcium channels and likely plays an important role in shaping the cardiac action potential and regulating Ca(2+) dynamics in the heart.


A pilot study identifying a potential plasma biomarker for determining EGFR mutations in exons 19 or 21 in lung cancer patients.
The most common type of lung cancer is non‑small cell lung cancer (NSCLC), which is frequently characterized by a mutation in the epidermal growth factor receptor (EGFR). Determining the presence of an EGFR mutation in lung cancer is important, as it determines the type of treatment that a patients will receive. Therefore, the aim of the present study was to apply high‑resolution metabolomics (HRM) using liquid chromatography‑mass spectrometry to identify significant compounds in human plasma samples obtained from South Korean NSCLC patients, as potential biomarkers for providing early detection and diagnosis of minimally‑invasive NSCLC. The metabolic differences between lung cancer patients without EGFR mutations were compared with patients harboring EGFR mutations. Univariate analysis was performed, with a false discovery rate of q=0.05, in order to identify significant metabolites between the two groups. In addition, hierarchical clustering analysis was performed to discriminate between the metabolic profiles of the two groups. Furthermore, the significant metabolites were identified and mapped using Mummichog software, in order to generate a potential metabolic network model. Using metabolome‑wide association studies, metabolic alterations were identified. Linoleic acid [303.23 m/z, (M+Na)+], 5‑methyl tetrahydrofolate [231.10 m/z, (M+2H)+] and N‑succinyl‑L‑glutamate‑5 semialdehyde [254.06 m/z, (M+Na)+], were observed to be elevated in patients harboring EGFR mutations, whereas tetradecanoyl carnitine [394.29 m/z, (M+Na)+] was observed to be reduced. This suggests that these compounds may be affected by the EGFR mutation. In conclusion, the present study identified four potential biomarkers in patients with EGFR mutations, using HRM combined with pathway analysis. These results may facilitate the development of novel diagnostic tools for EGFR mutation detection in patients with lung cancer.


miR-9 and miR-200 Regulate PDGFRβ-Mediated Endothelial Differentiation of Tumor Cells in Triple-Negative Breast Cancer.
Organization of cancer cells into endothelial-like cell-lined structures to support neovascularization and to fuel solid tumors is a hallmark of progression and poor outcome. In triple-negative breast cancer (TNBC), PDGFRβ has been identified as a key player of this process and is considered a promising target for breast cancer therapy. Thus, we aimed at investigating the role of miRNAs as a therapeutic approach to inhibit PDGFRβ-mediated vasculogenic properties of TNBC, focusing on miR-9 and miR-200. In MDA-MB-231 and MDA-MB-157 TNBC cell lines, miR-9 and miR-200 promoted and inhibited, respectively, the formation of vascular-like structures in vitro Induction of endogenous miR-9 expression, upon ligand-dependent stimulation of PDGFRβ signaling, promoted significant vascular sprouting of TNBC cells, in part, by direct repression of STARD13. Conversely, ectopic expression of miR-200 inhibited this sprouting by indirectly reducing the protein levels of PDGFRβ through the direct suppression of ZEB1. Notably, in vivo miR-9 inhibition or miR-200c restoration, through either the generation of MDA-MB-231-stable clones or peritumoral delivery in MDA-MB-231 xenografted mice, strongly decreased the number of vascular lacunae. Finally, IHC and immunofluorescence analyses in TNBC specimens indicated that PDGFRβ expression marked tumor cells engaged in vascular lacunae. In conclusion, our results demonstrate that miR-9 and miR-200 play opposite roles in the regulation of the vasculogenic ability of TNBC, acting as facilitator and suppressor of PDGFRβ, respectively. Moreover, our data support the possibility to therapeutically exploit miR-9 and miR-200 to inhibit the process of vascular lacunae formation in TNBC. Cancer Res; 76(18); 5562-72. ©2016 AACR.


Interaction between non-executive and executive directors in English National Health Service trust boards: an observational study.
National Health Service (NHS) trusts, which provide the majority of hospital and community health services to the English NHS, are increasingly adopting a 'public firm' model with a board consisting of executive directors who are trust employees and external non-executives chosen for their experience in a range of areas such as finance, health care and management. In this paper we compare the non-executive directors' roles and interests in, and contributions to, NHS trust boards' governance activities with those of executive directors; and examine non-executive directors' approach to their role in board meetings. Non-participant observations of three successive trust board meetings in eight NHS trusts (primary care trusts, foundation trusts and self-governing (non-foundation) trusts) in England in 2008-9. The observational data were analysed inductively to yield categories of behaviour reflecting the perlocutionary types of intervention which non-executive directors made in trust meetings. The observational data revealed six main perlocutionary types of questioning tactic used by non-executive directors to executive directors: supportive; lesson-seeking; diagnostic; options assessment; strategy seeking; and requesting further work. Non-executive board members' behaviours in holding the executive team to account at board meetings were variable. Non-executive directors were likely to contribute to finance-related discussions which suggests that they did see financial challenge as a key component of their role. The pattern of behaviours was more indicative of an active, strategic approach to governance than of passive monitoring or 'rubber-stamping'. Nevertheless, additional means of maintaining public accountability of NHS trusts may also be required.


Point-spread function associated with underwater imaging through a wavy air-water interface: theory and laboratory tank experiment.
The point-spread function needed for imaging underwater objects is theoretically derived and compared with experimental results. The theoretical development is based on the emergent-ray model, in which the Gram-Charlier series for the non-Gaussian probability-density function for emergent angles through a wavy water surface was assumed. To arrive at the point-spread model, we used a finite-element methodology with emergent-ray angular probability distributions as fundamental building functions. The model is in good agreement with the experiment for downwind conditions. A slight deviation between theory and experiment was observed for the crosswind case; this deviation may be caused by the possible interaction of standing waves with the original air-ruffled capillary waves that were not taken into account in the model.


CD24, a signal-transducing molecule expressed on human B cells, is a major surface antigen on small cell lung carcinomas.
Cell lines derived from human small cell carcinoma of the lung express high levels of a surface polypeptide termed the cluster-w4 antigen, which was previously identified as a potential target for toxin-based immunotherapy of lung cancer. We have cloned a complementary DNA encoding the cluster-w4 antigen from COS-1 fibroblasts transfected with a SW2 small cell carcinoma library, by panning with a mixture of the cluster-w4-specific monoclonal antibodies SWA11, SWA21, and SWA22. The sequence of the cluster-w4 complementary DNA encodes an unusually short (80-amino acid) protein identical to that recently reported for the leukocyte activation molecule CD24 except for a single valine-alanine substitution due to a single-base polymorphism within the region of the gene coding for the extracellular domain. Biochemical analyses of the cloned cluster-w4 antigen confirmed both the presence of the phosphatidylinositol tail and the extensive glycosylation reported for the CD24 molecule. Furthermore, the cloned cluster-w4 antigen expressed on COS cells was shown to react with a comprehensive panel of CD24-specific monoclonal antibodies, as assessed by indirect immunofluorescence staining. Northern blot hybridization indicated the presence of several transcript sizes for the cluster-w4 antigen that were greatly overexpressed in small cell carcinoma cell lines, compared with normal hemopoietic cells and CD24-positive cell lines. Southern blot hybridization of restriction digests of genomic DNA identified a complex pattern of bands consistent with either a complex gene structure containing many exons or the presence of a family of closely related genes.


Effect of Two Plant Species, Flax (Linum usitatissinum L.) and Tomato (Lycopersicon esculentum Mill.), on the Diversity of Soilborne Populations of Fluorescent Pseudomonads.
Suppression of soilborne disease by fluorescent pseudomonads may be inconsistent. Inefficient root colonization by the introduced bacteria is often responsible for this inconsistency. To better understand the bacterial traits involved in root colonization, the effect of two plant species, flax (Linum usitatissinum L.) and tomato (Lycopersicon esculentum Mill.), on the diversity of soilborne populations was assessed. Fluorescent pseudomonads were isolated from an uncultivated soil and from rhizosphere, rhizoplane, and root tissue of flax and tomato cultivated in the same soil. Species and biovars were identified by classical biochemical and physiological tests. The ability of bacterial isolates to assimilate 147 different organic compounds and to show three different enzyme activities was assessed to determine their intraspecific phenotypic diversity. Numerical analysis of these characteristics allowed the clustering of isolates showing a high level (87.8%) of similarity. On the whole, the populations isolated from soil were different from those isolated from plants with respect to their phenotypic characteristics. The difference in bacteria isolated from uncultivated soil and from root tissue of flax was particularly marked. The intensity of plant selection was more strongly expressed with flax than with tomato plants. The selection was, at least partly, plant specific. The use of 10 different substrates allowed us to discriminate between flax and tomato isolates. Pseudomonas fluorescens biovars II, III, and V and Pseudomonas putida biovar A and intermediate type were well distributed among the isolates from soil, rhizosphere, and rhizoplane. Most isolates from root tissue of flax and tomato belonged to P. putida bv. A and to P. fluorescens bv. II, respectively. Phenotypic characterization of bacterial isolates was well correlated with genotypic characterization based on repetitive extragenic palindromic PCR fingerprinting.


DLVO interactions of tungsten oxide and cobalt oxide surfaces measured with the colloidal probe technique.
We have investigated the DLVO surface forces of oxidized tungsten and cobalt surfaces using the atomic force microscope (AFM) colloidal probe technique. It was shown by X-ray photoelectron spectroscopy (XPS) and electrokinetic measurements that this model system is representative of industrial tungsten carbide (WC) and cobalt powders used in the production of hard metals. We found that the attractive van der Waals forces are well described by Hamaker constants, calculated from optical data for WO(3) and CoOOH. The repulsive electrostatic double layer forces between WO(3) surfaces increase with increasing pH due to an increasingly negative surface potential. This surface potential decreases with increasing ionic strength at pH 7.5. The electrostatic interaction between WO(3) and CoOOH is attractive at pH 10, suggesting a positively charged CoOOH surface.


Gold-catalyzed intramolecular cyclization/condensation sequence: synthesis of 1,2-dihydro[c][2,7]naphthyridines.
An efficient gold-catalyzed synthetic protocol for the construction of 1,2-dihydro[c][2,7]naphthyridines from 2-aminophenyl prop-2-yn-1-yl enaminones has been developed. In this organic transformation new C-C and C-N bond formations occur via 6-endo-dig cyclization and condensation. 1,2-Dihydro[c][2,7]naphthyridine derivatives were obtained in good to excellent yields in a one-pot synthesis at ambient temperature.


Pathologic Changes of Cone Photoreceptors in Eyes With Occult Macular Dystrophy.
Occult macular dystrophy (OMD) is an inherited retinal disease characterized by a progressive decrease of vision and appearance of normal fundus. To determine the pathologic features of OMD, we investigated the alternation of the photoreceptors using quantitative image analysis. We studied 22 eyes of 11 OMD patients. Three of them had a mutation (R45W) in RP1L1. The relative intensities of the ellipsoid zone in the spectral-domain optical coherence tomography (SD-OCT) images and the density of the cone photoreceptors in the adaptive optics (AO) fundus images of the OMD patients were compared to those of normal controls. The relative intensities of the ellipsoid zone in the SD-OCT images of patients with OMD were significantly lower (P < 0.001) by an average of 16% compared to that of the normal controls. Normal cone mosaics were not observed in the AO images of the macula in the eyes with OMD. The mean ± SD of cone density of the 9 OMD patients was 1970 ± 884 cells/mm2 at 2°, 1124 ± 483 cells/mm2 at 3°, and 1288 ± 715 cells/mm2 at 4° nasal to the fovea. The cone densities at 2°, 3°, and 4° nasal to the fovea of OMD were significantly lower than those of the normal controls (P < 0.001). A sparse array of cone photoreceptors with significantly reduced density of the macula is one of the morphologic features of OMD.


Improvements in the data quality of the Interferometric Monitor for Greenhouse Gases.
The Interferometric Monitor for Greenhouse Gases (IMG) operated aboard the polar-orbiting Advanced Earth Observing Satellite from October 1996 through June 1997. The IMG measured upwelling infrared radiance at fine spectral resolution. This paper identifies previously undocumented issues with IMG interferograms and describes procedures for correcting the majority of the affected data. In particular, single-sided interferograms should be used to avoid large noise bursts, and phase ambiguities must be resolved in uncalibrated spectra before radiometric calibration. The corrections are essential for studies that require accurately calibrated radiance spectra, including those that track atmospheric changes globally on decadal time scales.


Antioxidant properties of the essential oil of Eugenia caryophyllata and its antifungal activity against a large number of clinical Candida species.
Many essential oils are known to possess an antioxidant activity and antifungal properties and therefore they potentially act as antimycotic agents. Essential oil of clove (Eugenia caryophyllata) was isolated by hydrodistillation. The chemical composition of the essential oil was analysed by gas chromatography and gas chromatography/mass spectroscopy. The antioxidant effect of the tested oil was evaluated by measuring its 2,2-diphenyl-l-1-picrylhydrazil radical scavenging ability and the antiradical dose required to cause a 50% inhibition (IC50) was recorded. The antifungal activity of essential oils was evaluated against 53 human pathogenic yeasts using a disc paper diffusion method. Our results show that the major components present in the clove bund oil were eugenol (88.6%), eugenyl acetate (5.6%), beta-caryophyllene (1.4%) and 2-heptanone (0.9%). The tested essential oil exhibited a very strong radical scavenging activity (IC50 = 0.2 microg ml-1) when compared with the synthetic antioxidant (tert-butylated hydroxytoluene, IC50 = 11.5 microg ml-1). On the other hand, this species displayed an important antifungal effect against the tested strains. It is clear that clove oil shows powerful antifungal activity; and it can be used as an easily accessible source of natural antioxidants and in pharmaceutical applications.


Telomeres and endocrine dysfunction of the adrenal and GH/IGF-1 axes.
Telomeres, located at the end of linear chromosomes, are essential to maintain genomic stability. Telomere biology has recently emerged as an important player in the fields of ageing and disease. To maintain telomere length (TL) and reduce its degradation after mitosis, the telomerase enzyme complex is produced. Genetic, epigenetic, hormonal and environmental factors can regulate telomerase function. These include stress hormones such as cortisol and growth factors. The hypothalamic-pituitary-adrenal (HPA) axis has been evaluated in psychiatric diseases where hypercortisolism and oxidative stress are often present. Some researches have linked TL shortening to increases in stress-related cortisol, but others have not. The effects of cortisol on the telomere system are complex and may depend on the intensity and duration of exposure. On the other hand, low levels of IGF-1 are associated with inflammation and ageing-related diseases (ischaemic heart disease, congestive heart failure). Both IGF-1 and TL diminish with age and are positively and strongly correlated with each other. It is not clear whether this positive correlation reflects a single association or a cause-effect relationship. Further research will ideally investigate longitudinal changes in telomeres and both these hormonal axes. To our knowledge, TL dysfunction has not been described in either endogenous hypercortisolism (Cushing's syndrome) or acromegaly where excessive amounts of GH and consequently IGF-1 are produced. This review focuses on the possible relationships between telomere dysfunction and the hypothalamic-pituitary-adrenal (HPA) axis and GH-IGF-1 system.


Determining chromatin architecture with Micro Capture-C.
Micro Capture-C (MCC) is a chromatin conformation capture (3C) method for visualizing reproducible three-dimensional contacts of specified regions of the genome at base pair resolution. These methods are an established family of techniques that use proximity ligation to assay the topology of chromatin. MCC can generate data at substantially higher resolution than previous techniques through multiple refinements of the 3C method. Using a sequence agnostic nuclease, the maintenance of cellular integrity and full sequencing of the ligation junctions, MCC achieves subnucleosomal levels of resolution, which can be used to reveal transcription factor binding sites analogous to DNAse I footprinting. Gene dense regions, close-range enhancer-promoter contacts, individual enhancers within super-enhancers and multiple other types of loci or regulatory regions that were previously challenging to assay with conventional 3C techniques, are readily observed using MCC. MCC requires training in common molecular biology techniques and bioinformatics to perform the experiment and analyze the data. The protocol can be expected to be completed in a 3 week timeframe for experienced molecular biologists.


All-fiber noninterferometric narrow-transmission-bandpass filter.
In-fiber mode engineering based on the combination of Bragg and long-period gratings (LPGs) permits the implementation of noninterferometric transmission filters with narrow passbands using standard single-mode fiber. The design of the bandpass filter is based on the coupling between propagating and counterpropagating cladding modes in two fiber Bragg gratings. A LPG located between the Bragg gratings transfers power from the input fundamental mode to a specific cladding mode and recouples the filtered signal to the output fundamental mode. The filter produces a series of narrow passbands of about 30 pm linewidth with a maximum transmittance above 60%, 20 dB isolation, and passband separation of about 1 nm, each corresponding to the contribution of a different cladding mode.


Automatic identification of chemical moieties.
In recent years, the prediction of quantum mechanical observables with machine learning methods has become increasingly popular. Message-passing neural networks (MPNNs) solve this task by constructing atomic representations, from which the properties of interest are predicted. Here, we introduce a method to automatically identify chemical moieties (molecular building blocks) from such representations, enabling a variety of applications beyond property prediction, which otherwise rely on expert knowledge. The required representation can either be provided by a pretrained MPNN, or be learned from scratch using only structural information. Beyond the data-driven design of molecular fingerprints, the versatility of our approach is demonstrated by enabling the selection of representative entries in chemical databases, the automatic construction of coarse-grained force fields, as well as the identification of reaction coordinates.


Young children's video/computer game use: relations with school performance and behavior.
This study examined the amount and content of children's video game playing in relation with behavioral and academic outcomes. Relationships among playing context, child gender, and parental monitoring were explored. Data were obtained through parent report of child's game play, behavior, and school performance. Results revealed that time spent playing games was related positively to aggression and negatively to school competence. Violent content was correlated positively and educational content negatively with attention problems. Educational games were related to good academic achievement. Results suggest violent games, and a large amount of game play, are related to troublesome behavioral and academic outcomes, but educational games may be related to positive outcomes. Neither gender nor parental monitoring emerged as significant moderators of these effects.


An update on the modern management of paediatric uveitis.
Uveitis in children and young people (CYP) is often painless, chronic and persistent. It is an often silent blinding condition. Uveitis can be isolated or develop as a manifestation of a systemic disease. Due to the symptomless nature, it can present late with advanced ocular comorbidities such as band keratopathy, hypotony, cataracts. Inadequate control of the eye inflammation can result in permanent and severe ocular complications, structural damage and visual loss. One of the most common systemic associations is juvenile idiopathic arthritis where uveitis has a cumulative incidence of approximately 10%-14% (though wide variation in incidence is reported) after 5 years. Appropriately targeted uveitis screening is recommended to continue for at least 7 years in some subgroups. Paediatric uveitis poses multiple diagnostic and therapeutic challenges. Clinical manifestation and disease course may differ significantly from adult-onset uveitis. However, some CYP are still managed by adult specialists alone, without the opportunity for the prompt use of National Health Service England approved therapy. Optimal management of paediatric uveitis requires a multidisciplinary approach involving coordinated working of different specialities and healthcare professionals. This article highlights the evidence-based practice for the contemporary management of paediatric uveitis to readers in different specialities who may come across this condition. It raises awareness of early systemic treatment aiming to achieve early and complete disease inactivity thereby improving the chances of a long-term positive outcome.


Concrete versus abstract forms of social concept: an fMRI comparison of knowledge about people versus social terms.
The anterior temporal lobes (ATLs) play a key role in conceptual knowledge representation. The hub-and-spoke theory suggests that the contribution of the ATLs to semantic representation is (a) transmodal, i.e. integrating information from multiple sensorimotor and verbal modalities, and (b) pan-categorical, representing concepts from all categories. Another literature, however, suggests that this region's responses are modality- and category-selective; prominent examples include category selectivity for socially relevant concepts and face recognition. The predictions of each approach have never been directly compared. We used data from three studies to compare category-selective responses within the ATLs. Study 1 compared ATL responses to famous people versus another conceptual category (landmarks) from visual versus auditory inputs. Study 2 compared ATL responses to famous people from pictorial and written word inputs. Study 3 compared ATL responses to a different kind of socially relevant stimuli, namely abstract non-person-related words, in order to ascertain whether ATL subregions are engaged for social concepts more generally or only for person-related knowledge. Across all three studies a dominant bilateral ventral ATL cluster responded to all categories in all modalities. Anterior to this 'pan-category' transmodal region, a second cluster responded more weakly overall yet selectively for people, but did so equally for spoken names and faces (Study 1). A third region in the anterior superior temporal gyrus responded selectively to abstract socially relevant words (Study 3), but did not respond to concrete socially relevant words (i.e. written names; Study 2). These findings can be accommodated by the graded hub-and-spoke model of concept representation. On this view, the ventral ATL is the centre point of a bilateral ATL hub, which contributes to conceptual representation through transmodal distillation of information arising from multiple modality-specific association cortices. Partial specialization occurs across the graded ATL hub as a consequence of gradedly differential connectivity across the region.This article is part of the theme issue 'Varieties of abstract concepts: development, use and representation in the brain'.


Identification of protein disulfide isomerase as a cardiomyocyte survival factor in ischemic cardiomyopathy.
The aim of the study was to analyze the molecular mechanisms activated during postinfarction remodeling in human hearts. The molecular mechanisms of initial response to ischemic insult in the heart and the pathways involved in compensation and remodeling are still largely unknown. Up-regulation or down-regulation of gene expression in the human viable peri-infarct (vs. remote) myocardial region was investigated by complementary deoxyribonucleic acid array technology and confirmed at a single-gene/protein level with reverse transcriptase polymerase chain reaction and immunohistochemistry. An in vitro model of cardiomyocyte hypoxia in HL1 cells was used to validate anti-apoptotic effects of the candidate gene/protein and to assess the associated downstream cascade. Finally, a mouse model of myocardial infarction was used to test the in vivo effects of exogenous transfection with the candidate gene/protein. Protein disulfide isomerase (PDI), a member of the unfolded protein response, is 3-fold up-regulated in the viable peri-infarct myocardial region, and in a postmortem model, its expression is significantly inversely correlated with apoptotic rate and with presence of heart failure (HF) and biventricular dilatation. Induced PDI expression in HL1 cells conferred protection from hypoxia-induced apoptosis. Adenoviral-mediated PDI gene transfer to the mouse heart resulted in 2.5-fold smaller infarct size, significantly reduced cardiomyocyte apoptosis in the peri-infarct region, and smaller left ventricular end-diastolic diameter versus mice treated with a transgene-null adenoviral vector. These results suggest that PDI promotes survival after ischemic damage and that zinc-superoxide dismutase is one of the PDI molecular targets. Pharmacological modulation of this pathway might prove useful for future prevention and treatment of HF.


Inhibition of NADPH oxidase 4 induces apoptosis in malignant mesothelioma: Role of reactive oxygen species.
Malignant pleural mesothelioma (MPM) is an aggressive tumor that is characterized by dysregulated growth and resistance to apoptosis. Reactive oxygen species (ROS)-generating NADPH oxidase (Nox) family enzymes have been suggested to be involved in neoplastic proliferation. Both the antioxidant N-acetylcysteine (NAC) and the inhibitor of flavoprotein-dependent oxidase, diphenylene iodonium (DPI), inhibited the cell viability of MPM cells in a dose-dependent manner. To examine whether Nox-mediated ROS generation confers antiapoptotic activity and thus a growth advantage to MPM cells, we analyzed the mRNA expression of Nox family members using quantitative RT-PCR in 7 MPM cell lines and a normal mesothelial cell line. Nox4 mRNA was expressed in all of the examined MPM cell lines, whereas little or no Nox2, Nox3 and Nox5 mRNA expression was detected. In 2 MPM cell lines, Nox4 mRNA expression was significantly higher than that in a normal mesothelial cell line. siRNAs targeting Nox4 suppressed ROS generation and cell viability in the MPM cell lines. In addition, DPI treatment and knockdown of Nox4 attenuated phosphorylation of AKT and ERK. Taken together, our results indicate that Nox4-mediated ROS, at least in part, transmit cell survival signals and their depletion leads to apoptosis, thus highlighting the Nox4-ROS-AKT signaling pathway as a potential therapeutic target for MPM treatment.


Ultrasound imaging for regional anesthesia in infants, children, and adolescents: a review of current literature and its application in the practice of extremity and trunk blocks.
The use of ultrasound guidance has provided an opportunity to perform many peripheral nerve blocks that would have been difficult to perform in children based on pure landmark techniques due to the potential for injection into contiguous sensitive vascular areas. This review article provides the readers with techniques on ultrasound-guided peripheral nerve blocks of the extremities and trunk with currently available literature to substantiate the available evidence for the use of these techniques. Ultrasound images of the blocks with corresponding line diagrams to demonstrate the placement of the ultrasound probe have been provided for all the relevant nerve blocks in children. The authors hope that this review will stimulate further research into ultrasound-guided regional anesthesia in infants, children, and adolescents and stimulate more randomized controlled trials to provide a greater understanding of the anatomy and physiology of regional anesthesia in pediatrics.


Reducing wrong patient selection errors: exploring the design space of user interface techniques.
Wrong patient selection errors are a major issue for patient safety; from ordering medication to performing surgery, the stakes are high. Widespread adoption of Electronic Health Record (EHR) and Computerized Provider Order Entry (CPOE) systems makes patient selection using a computer screen a frequent task for clinicians. Careful design of the user interface can help mitigate the problem by helping providers recall their patients' identities, accurately select their names, and spot errors before orders are submitted. We propose a catalog of twenty seven distinct user interface techniques, organized according to a task analysis. An associated video demonstrates eighteen of those techniques. EHR designers who consider a wider range of human-computer interaction techniques could reduce selection errors, but verification of efficacy is still needed.


Injury reactive myelin/oligodendrocyte-derived axon growth inhibition in the adult mammalian central nervous system.
Myelin inhibition is considered a constitutive, static, repulsive barrier not reactive to a central nervous system (CNS) lesion. However, recent evidence underlines the existence of considerable add-on axon growth inhibition upon CNS injury. This postlesional, reactive myelin/oligodendrocyte-derived inhibition will require the development of novel screening approaches and therapeutic reagents to promote axonal regeneration.


A population-based study on healthcare-seeking behaviour of persons with symptoms of respiratory and gastrointestinal-related infections in Hong Kong.
Studies on healthcare-seeking behaviour usually adopted a patient care perspective, or restricted to specific disease conditions. However, pre-diagnosis symptoms may be more relevant to healthcare-seeking behaviour from a patient perspective. We described healthcare-seeking behaviours by specific symptoms related to respiratory and gastrointestinal-related infections. We conducted a longitudinal population-based telephone survey in Hong Kong. We collected data on healthcare-seeking behaviour specific to symptoms of respiratory and gastrointestinal-related infections and also associated demographic factors. We performed descriptive analyses and estimated the proportion of participants who sought medical consultation, types of services utilized and duration from symptom onset to healthcare seeking, by different age groups. Post-stratification was used to compensate non-response and multiple imputation to handle missing and right-censored data. We recruited 2564 participants who reported a total of 4370 illness episodes and 7914 symptoms. Fatigue was the most frequently reported symptom, followed by headache and runny nose, with 30-day incidence rate of 9.1, 7.7, and 7.7% respectively. 78% of the participants who had fever sought medical consultation, followed by those with rash (60%) and shortness of breath (58%). Older adults (aged ≥55y) who had symptoms including fever, sore throat, and headache had a significantly higher consultation rate comparing to the other age groups. The 30-day incidence rates of influenza-like illness (ILI) and acute respiratory illness (ARI) were 0.8 and 7.2% respectively, and the consultation rates among these participants were 91 and 64%. Private general practitioner clinics was the main service utilized by participants for most of the symptoms considered, especially those related to acute illness such as fever, diarrhoea and vomiting. Chinese medicine clinics were mostly likely to be visited by participants with low back pain, myalgia and fatigue. Among participants who have sought medical services, most were within 3 days of symptom onset. Healthcare-seeking behaviour were different by symptoms and age. Characterization of these patterns provides crucial parameters for estimating the full burden of common infectious diseases from facility-based surveillance system, for planning and allocation of healthcare resources.


Genomic epidemiology and evolutionary dynamics of respiratory syncytial virus group B in Kilifi, Kenya, 2015-17.
Respiratory syncytial virus (RSV) circulates worldwide, occurring seasonally in communities, and is a leading cause of acute respiratory illness in young children. There is paucity of genomic data from purposively sampled populations by which to investigate evolutionary dynamics and transmission patterns of RSV. Here we present an analysis of 295 RSV group B (RSVB) genomes from Kilifi, coastal Kenya, sampled from individuals seeking outpatient care in nine health facilities across a defined geographical area (∼890 km2), over two RSV epidemics between 2015 and 2017. RSVB diversity was characterized by multiple virus introductions into the area and co-circulation of distinct genetic clusters, which transmitted and diversified locally with varying frequency. Increase in relative genetic diversity paralleled seasonal virus incidence. Importantly, we identified a cluster of viruses that emerged in the 2016/17 epidemic, carrying distinct amino-acid signatures including a novel nonsynonymous change (K68Q) in antigenic site ∅ in the Fusion protein. RSVB diversity was additionally marked by signature nonsynonymous substitutions that were unique to particular genomic clusters, some under diversifying selection. Our findings provide insights into recent evolutionary and epidemiological behaviors of RSVB, and highlight possible emergence of a novel antigenic variant, which has implications on current prophylactic strategies in development.


Psychological and physiological responses following repeated peer death.
Undergraduates at a university in the United States were exposed - directly and indirectly - to 14 peer deaths during one academic year. We examined how individual and social factors were associated with psychological (e.g., anxiety, depression, somatization) and physiological (i.e., cortisol) distress responses following this unexpected and repeated experience with loss. Two to three months after the final peer death, respondents (N = 122, 61% female, 18-23 years, M = 20.13, SD = 1.14) reported prior adverse experiences, degree of closeness with the deceased, acute responses to the peer deaths, ongoing distress responses, social support, support seeking, and media viewing. A subset (n = 24) returned hair samples for evaluation of cortisol responses during the previous 3 months. Ongoing psychological distress was associated with a) prior interpersonal trauma, b) fewer social supports, and c) media exposure to news of the deaths (p's<.05). Participants who had no prior bereavements showed, on average, high cortisol (>25 p/mg) compared to individuals with one or two prior bereavement experiences (who were, on average, within the normal range, 10 to 25 p/mg) (p<.05). Only 8% of the sample utilized available university psychological or physical health resources and support groups. Limited research has examined the psychological and physiological impact of exposure to chronic, repeated peer loss, despite the fact that there are groups of individuals (e.g., police, military soldiers) that routinely face such exposures. Prior adversity appears to play a role in shaping psychological and physiological responses to repeated loss. This topic warrants further research given the health implications of repeated loss for individuals in high-risk occupations and university settings.


Epigenetic inactivation of tumor suppressor SFRP2 and point mutation in KRAS proto-oncogene in fistula-associated mucinous type anal adenocarcinoma: report of two cases.
The secreted frizzled-related proteins (SFRPs) genes are unmethylated in normal colorectal mucosa tissue but aberrant methylation profiles can be detected in colorectal cancer (CRC), adenomas, and in aberrant crypt foci. The aim of the current study was to clarify whether SFRP2 methylation and K-ras structural mutation in fecal DNA can be found in stool and tumoral tissues of individuals with fistula-associated mucinous type anal adenocarcinomas (MTAA).Two man patients (68 and 56 years old) were treated for anorectal fistula in the surgical department. Patients were evaluated for clinical findings, tumoural tissue samples were examined histopathologically and DNA from fecal and tumoral tissue samples were isolated. K-ras mutation and promoter hypermethylation of SFRP2 gene in tumoral tissues were assessed by methylation-specific PCR based stripAssay hybridisation technique (Me-PCR) and compared to the healthy controls. Fecal and tumoural tissue samples from both patients were found to be fully hypermethylated profiles for SFRP2 gene and combined point mutations were detected in codon 12 and 13 of K-ras proto-oncogene. The current results showed that the combined effects of somatic mutations in K-ras and epigenetic alterations in SFRP2 genes may play an active role in the development of mucinous type anal adenocarcinoma.


Self-referenced luminescence thermometry with Sm(3+) doped TiO2 nanoparticles.
The performance of Sm(3+) doped TiO2 nanoparticles for luminescence temperature sensing was tested over a temperature range from room to 110 °C. The Sm(3+) ions were incorporated into TiO2 nanocrystals using hydrolytic sol-gel route. Microstructural characterization of the obtained material was performed using transmission electron microscopy and x-ray diffraction measurements. Luminescence emission spectra of Sm(3+) doped TiO2 nanoparticles consists of two distinct spectral regions: the high energy region associated with the trap emission of the TiO2 host, and the low energy region with well-resolved emission peaks of the Sm(3+) ions. The ratio between Sm(3+) emission and TiO2 trap emission shows strong temperature dependence, and is tested for temperature sensing. The relative sensor sensitivity was found to be higher than 1% °C(-1) over given temperature range with the maximum value of 10.54% °C(-1) at 57.5 °C. Lifetime data derived from the Sm(3+) emission decay revealed that time-resolved measurements provide comparable quality of temperature sensing as corresponding ratiometric measurements, with a maximum relative sensitivity of 10.14% °C(-1) at 66.5 °C.


Mid-infrared-pumped quantum cascade structure for high-sensitive terahertz detection.
Based on multiple quantum wells, we design a pumping-detection quantum cascade structure for the detection of terahertz (THz) radiation. In the structure, carriers are first pumped by a mid-infrared (MIR) laser to an excited state, to get enough energy space for the following fast longitudinal optical (LO) phonon extraction. Within the LO-phonon extraction stair, an absorption well is designed for THz detection. Due to the establishment of LO-phonon stair extractor, carriers transport between quantum wells in picosecond range and a high responsivity for THz absorption can be obtained. We also find that doping in both MIR active well and extractor region is significant for high-speed response of the THz detection. Our design is expected to extend the high-sensitive detection of a quantum cascade photodetector from middle wave of MIR to THz region.


Biochemical, Metabolomic, and Genetic Analyses of Dephospho Coenzyme A Kinase Involved in Coenzyme A Biosynthesis in the Human Enteric Parasite Entamoeba histolytica.
Coenzyme A (CoA) is an essential cofactor for numerous cellular reactions in all living organisms. In the protozoan parasite Entamoeba histolytica, CoA is synthesized in a pathway consisting of four enzymes with dephospho-CoA kinase (DPCK) catalyzing the last step. However, the metabolic and physiological roles of E. histolytica DPCK remain elusive. In this study, we took biochemical, reverse genetic, and metabolomic approaches to elucidate role of DPCK in E. histolytica. The E. histolytica genome encodes two DPCK isotypes (EhDPCK1 and EhDPCK2). Epigenetic gene silencing of Ehdpck1 and Ehdpck2 caused significant reduction of DPCK activity, intracellular CoA concentrations, and also led to growth retardation in vitro, suggesting importance of DPCK for CoA synthesis and proliferation. Furthermore, metabolomic analysis showed that suppression of Ehdpck gene expression also caused decrease in the level of acetyl-CoA, and metabolites involved in amino acid, glycogen, hexosamine, nucleic acid metabolisms, chitin, and polyamine biosynthesis. The kinetic properties of E. histolytica and human DPCK showed remarkable differences, e.g., the Km values of E. histolytica and human DPCK were 58-114 and 5.2 μM toward dephospho-CoA and 15-20 and 192 μM for ATP, respectively. Phylogenetic analysis also supported the uniqueness of the amebic enzyme compared to the human counterpart. These biochemical, evolutionary features, and physiological importance of EhDPCKs indicate that EhDPCK represents the rational target for the development of anti-amebic agents.


Effect of Dietary Sodium on Blood Pressure: A Crossover Trial.
Dietary sodium recommendations are debated partly due to variable blood pressure (BP) response to sodium intake. Furthermore, the BP effect of dietary sodium among individuals taking antihypertensive medications is understudied. To examine the distribution of within-individual BP response to dietary sodium, the difference in BP between individuals allocated to consume a high- or low-sodium diet first, and whether these varied according to baseline BP and antihypertensive medication use. Prospectively allocated diet order with crossover in community-based participants enrolled between April 2021 and February 2023 in 2 US cities. A total of 213 individuals aged 50 to 75 years, including those with normotension (25%), controlled hypertension (20%), uncontrolled hypertension (31%), and untreated hypertension (25%), attended a baseline visit while consuming their usual diet, then completed 1-week high- and low-sodium diets. High-sodium (approximately 2200 mg sodium added daily to usual diet) and low-sodium (approximately 500 mg daily total) diets. Average 24-hour ambulatory systolic and diastolic BP, mean arterial pressure, and pulse pressure. Among the 213 participants who completed both high- and low-sodium diet visits, the median age was 61 years, 65% were female and 64% were Black. While consuming usual, high-sodium, and low-sodium diets, participants' median systolic BP measures were 125, 126, and 119 mm Hg, respectively. The median within-individual change in mean arterial pressure between high- and low-sodium diets was 4 mm Hg (IQR, 0-8 mm Hg; P < .001), which did not significantly differ by hypertension status. Compared with the high-sodium diet, the low-sodium diet induced a decline in mean arterial pressure in 73.4% of individuals. The commonly used threshold of a 5 mm Hg or greater decline in mean arterial pressure between a high-sodium and a low-sodium diet classified 46% of individuals as "salt sensitive." At the end of the first dietary intervention week, the mean systolic BP difference between individuals allocated to a high-sodium vs a low-sodium diet was 8 mm Hg (95% CI, 4-11 mm Hg; P < .001), which was mostly similar across subgroups of age, sex, race, hypertension, baseline BP, diabetes, and body mass index. Adverse events were mild, reported by 9.9% and 8.0% of individuals while consuming the high- and low-sodium diets, respectively. Dietary sodium reduction significantly lowered BP in the majority of middle-aged to elderly adults. The decline in BP from a high- to low-sodium diet was independent of hypertension status and antihypertensive medication use, was generally consistent across subgroups, and did not result in excess adverse events. ClinicalTrials.gov Identifier: NCT04258332.


GAP activity of the Yersinia YopE cytotoxin specifically targets the Rho pathway: a mechanism for disruption of actin microfilament structure.
The YopE cytotoxin of Yersinia pseudotuberculosis is an essential virulence determinant that is injected into the eukaryotic target cell via a plasmid-encoded type III secretion system. Injection of YopE into eukaryotic cells induces depolymerization of actin stress fibres. Here, we show that YopE exhibits a GTPase-activating protein (GAP) activity and that the presence of YopE stimulates downregulation of Rho, Rac and Cdc42 activity. YopE has an arginine finger motif showing homology with those found in other GAP proteins. Exchange of arginine 144 with alanine, located in this arginine finger motif, results in an inactive form of YopE that can no longer stimulate GTP hydrolysis by the GTPase. Furthermore, a yopE(R144A) mutant is unable to induce cytotoxicity on cultured HeLa cells in contrast to the corresponding wild-type strain. Expression of wild-type YopE in cells of Saccharomyces cerevisiae inhibits growth, while in contrast, expression of the inactive form of YopE, YopE(R144A), does not affect the yeast cells. Co-expression of proteins belonging to the Rho1 pathway of yeast, Rho1, Rom2p, Bck1 and Ste20, suppressed the growth phenotype of YopE in yeast cells. These results provide evidence that YopE exhibits a GAP activity to inactivate RhoGTPases, leading to depolymerization of the actin stress fibres in eukaryotic cells and growth inhibition in yeast.


Internet-delivered mental health treatment systems in Scandinavia - A usability evaluation.
Mental health problems are a major public health concern worldwide. Approximately 50% of the population will experience mental problems during their life. Traditional treatment is based on psychopharmacotherapy or psychotherapy, with face-to-face interaction between the patient and the therapist. New technologies such as Internet-delivered treatments are seen as an opportunity to offer more scalable and cost-efficient treatments in the field of mental health. Despite the growing interest and new evidence supporting the effect of Internet-delivered treatments is it remarkably little research on how the technology and the usability of Internet-delivered treatment programs affects the treatment. In this paper, we propose a set of evaluation criteria for evaluating the usability and the responsive design of Internet-delivered treatment systems. By our knowledge we are the first to include usability and universal design principles in the evaluation of Internet-delivered treatment systems. Our findings indicate that despite the good treatment results and proven clinical effects, the systems in general have several issues regarding usability, universal design and outdated technology. Based on our findings we propose that there should be established guidelines for testing the usability and technology of Internet-delivered treatment systems.


ActiveWV: a systematic approach to developing a physical activity plan for West Virginia.
Modeled after the National Physical Activity Plan (NPAP), ActiveWV 2015: The West Virginia Physical Activity Plan was developed to provide strategic direction for physical activity promotion within the state. The purpose of this manuscript is to describe the systematic approach taken in developing ActiveWV. Plan development began with establishing capacity and leadership among key stakeholders representing all societal sectors. A multiphase, statewide decision-making process allowed for input across sectors and geographic regions. The process results identified five priority areas that served as the conceptual framework for ActiveWV. Sector teams, comprised of key organization stakeholders across the eight sectors, finalized the sector-specific strategies and tactics using the NPAP evidence-based recommendations, results from a formalized strategic process, and the teams' expertise and experience. ActiveWV was officially released on January 19, 2012 at the State Capitol in Charleston, West Virginia. Community events throughout the state surrounded the release and celebrated West Virginia Physical Activity Day. Ongoing implementation and dissemination efforts are underway at state and local levels. As the NPAP calls for states and communities to develop plans that meet the needs of their particular context, other states may find the lessons learned from West Virginia helpful in the development process.


Formation of abdominal cerebrospinal fluid pseudocyst.
A 63-year-old man who underwent insertion of a lumboperitoneal shunt developed gait disturbance. He had undergone surgery for gastric cancer 7 years and for ileus 5 years previously. Head computed tomography (CT) revealed enlargement of the ventricles. Abdominal CT revealed a cyst in the abdominal region and the distal segment of the peritoneal shunt tube located within the cyst. Laparotomy revealed the cyst located between the small intestine, colon, and peritoneum. The anterior wall of the cyst was excised. The distal segment of the peritoneal shunt tube was replaced in the rectovesical pouch. Histological examination showed that the cyst wall consisted of inner fibrous tissue and outer fat tissue without epithelial lining, and invasion of lymphocytes. The diagnosis was pseudocyst. Only 29 cases of abdominal cerebrospinal fluid (CSF) pseudocysts have been reported in adults. Although the mechanism underlying the formation of abdominal pseudocyst remains to be clarified, several predisposing factors for cyst formation have been reported including changes in absorption of CSF due to inflammation or infection, peritoneal adhesions due to previous abdominal surgery, and increase in the protein content of the CSF. In our case, the medical history and histological features of the cyst wall indicated that formation of the abdominal pseudocyst was associated with previous surgery or inflammatory reaction.


Collaboration and Shared Decision-Making Between Patients and Clinicians in Preventive Health Care Decisions and US Preventive Services Task Force Recommendations.
The US Preventive Services Task Force (USPSTF) works to improve the health of people nationwide by making evidence-based recommendations for preventive services. Patient-centered care is a core value in US health care. Shared decision-making (SDM), in which patients and clinicians make health decisions together, ensures patients' rights to be informed and involved in preventive care decisions and that these decisions are patient-centered. SDM has a role across the spectrum of USPSTF recommendations. For A or B recommendations (judged by the USPSTF to have high or moderate certainty of a moderate or substantial net benefit at the population level), SDM allows individual patients to decide whether to accept such services based on their personal values and preferences. For C recommendations (indicating at least moderate certainty of a small net benefit at the population level), SDM is critical for individual patients to decide whether the net benefit for them is worthwhile. For D recommendations (reflecting at least moderate certainty of a zero or negative net benefit) or I statements (low certainty of net benefit), clinicians should be prepared to discuss these services if patients ask. More evidence is needed to determine if, in addition to promoting patient-centeredness, SDM reduces inequities in preventive care, as well as to define new strategies to find time for discussion of preventive services in primary care.


Forces shaping the antibiotic resistome.
Antibiotic resistance has become a problem of global scale. Resistance arises through mutation or through the acquisition of resistance gene(s) from other bacteria in a process called horizontal gene transfer (HGT). While HGT is recognized as an important factor in the dissemination of resistance genes in clinical pathogens, its role in the environment has been called into question by a recent study published in Nature. The authors found little evidence of HGT in soil using a culture-independent functional metagenomics approach, which is in contrast to previous work from the same lab showing HGT between the environment and human microbiome. While surprising at face value, these results may be explained by the lack of selective pressure in the environment studied. Importantly, this work suggests the need for careful monitoring of environmental antibiotic pollution and stringent antibiotic stewardship in the fight against resistance.


AGS3, an alpha(1-3)glucan synthase gene family member of Aspergillus fumigatus, modulates mycelium growth in the lung of experimentally infected mice.
The cell wall of human fungal pathogen Aspergillus fumigatus protects the fungus against threats from environment and interacts with the host immune system. Alpha(1-3)glucan is the major polysaccharide of Aspergillus fumigatus cell wall, and it has been shown to contribute to the virulence of diverse fungal pathogens. In A. fumigatus, three putative alpha(1-3)glucan synthase genes AGS1, AGS2 and AGS3 have been identified. AGS1 is responsible for cell wall alpha(1-3)glucan biosynthesis, but strains with deletions of either AGS1 or AGS2 are not defective in virulence [Beauvais, A., Maubon, D., Park, S., Morelle, W., Tanguy, M., Huerre, M., Perlin, D.S., Latgé, J. P., 2005. Two alpha(1-3) glucan synthases with different functions in Aspergillus fumigatus. Appl. Environ. Microbiol. 71, 1531-1538]. In contrast, we present evidence that AGS3 is also responsible for cell wall alpha(1-3)glucan biosynthesis and can modulate the virulence of A. fumigatus. An AGS3 deletion strain was found to produce faster and more robust disease than the parental strain in an experimental mouse model of aspergillosis. The apparent hyper-virulence in the AGS3-deleted mutant was correlated with an increased melanin content of the conidial cell wall, a better resistance to reactive oxygen species and a quicker germination rate. These results suggest an indirect role for AGS3 in virulence through an adaptive mechanism.


Targeted inactivation and identification of targets of the Gli2a transcription factor in the zebrafish.
Hedgehog (Hh) signaling is mediated by the Gli transcription factors and, in the zebrafish, plays an important role in patterning both the neural tube and myotome. Using a null allele of the gli2a gene induced by targeted mutagenesis, we show that Gli2a is completely dispensable in the fish but acts redundantly with Gli1 to regulate expression of known Hh targets, such as ptch2, prdm1a and eng2a, in the myotome and neural tube. To identify novel targets of Hh signaling, we performed chromatin immunoprecipitation sequencing (ChIP-seq) of whole embryo extracts. Samples were significantly enriched for 192 genomic regions, some of which are associated with four known Hh target genes, ptch1, ptch2, gli1 and olig2. Sequence analysis of these regions reveals a high level of conservation of Gli-binding sites from fish to mammals in some, but not all, cases. Expression analysis of other transcription units that are closely associated with peaks identified several putative targets not previously implicated as Hh targets, including myl10, hnmt, lrp4, efemp2, fras1, quo, and lamc1. Each of these genes shows loss of, or reduced expression in, embryos homozygous for an antimorphic allele of gli2a, you-too (yot), consistent with their being direct targets of Gli2a.


Olive leaf extract attenuates cardiac, hepatic, and metabolic changes in high carbohydrate-, high fat-fed rats.
Olive oil, an important component of the Mediterranean diet, produces cardioprotective effects, probably due to both oleic acid and the polyphenols such as oleuropein and hydroxytyrosol. Our aim in this study was to assess whether a polyphenol-enriched extract from the leaves of Olea europaea L. with oleuropein as the major component attenuated the cardiovascular, hepatic, and metabolic signs of a high-carbohydrate, high-fat (HCHF) diet (carbohydrate, 52%; fat, 24%, 25% fructose in drinking water) in rats. Male Wistar rats were fed either a cornstarch diet (CS) or a HCHF diet for a total of 16 wk. Diets of the treatment groups [CS+olive leaf extract (OLE) and HCHF+OLE] were supplemented with 3% OLE after 8 wk of being fed their respective CS or HCHF diets for a further 8 wk. After 16 wk, HCHF rats developed signs of metabolic syndrome, including elevated abdominal and hepatic fat deposition, collagen deposition in heart and liver, cardiac stiffness, and oxidative stress markers (plasma malondialdehyde and uric acid concentrations), with diminished aortic ring reactivity, abnormal plasma lipid profile, impaired glucose tolerance, and hypertension. Compared with HCHF rats, those in the HCHF+OLE group had improved or normalized cardiovascular, hepatic, and metabolic signs with the exception of elevated blood pressure. These results strongly suggest that an OLE containing polyphenols such as oleuropein and hydroxytyrosol reverses the chronic inflammation and oxidative stress that induces the cardiovascular, hepatic, and metabolic symptoms in this rat model of diet-induced obesity and diabetes without changing blood pressure.