Effect of chronic alcohol consumption on Hepatic SIRT1 and PGC-1alpha in rats.
The nuclear genes, NAD-dependent deacetylase Sirtuis 1 (SIRT1) and the peroxisome proliferator-activated receptor-gamma coactivator1alpha (PGC-1alpha) are regulators of energy metabolism. Here, we studied the role of alcohol consumption in expression of these sensing molecules. Alcohol significantly reduced hepatic SIRT1 mRNA by 50% and PGC-1alpha mRNA by 46% and it significantly inhibited the protein expression of SIRT1 and PGC-1alpha, while the transcription factor PPAR-gamma remained unchanged. However, when the lipid composition of the alcohol diet was changed by replacing long-chain triglycerides (LCT) with medium chain triglycerides (MCT), SIRT1 and PGC-1alpha mRNA were restored to near control levels. This study demonstrates that alcohol reduces key energy sensing proteins and that replacement of LCT by MCT affects the transcription of these genes. Since there is a pathophysiological link between SIRT1 and PGC-1alpha and mitochondrial energy, the implication of the study is that mitochondrial dysfunction due to alcohol abuse can be treated by dietary modifications.


Sustained impact of a rapid microarray-based assay with antimicrobial stewardship interventions on optimizing therapy in patients with Gram-positive bacteraemia.
To compare antibiotic optimization and outcomes of patients before implementation of the Verigene Gram-Positive Blood Culture (Verigene BC-GP) nucleic acid microarray assay to after implementation with antimicrobial stewardship (AS) interventions and after discontinuation of AS interventions. A retrospective pre-post-post quasi-experimental study was conducted to compare the three periods. AS interventions consisted of real-time guidance to clinicians on antibiotic selection. The primary outcome was median time from Gram stain to optimal therapy. Secondary outcomes included median time to effective therapy, median duration of therapy for contaminant organisms, median length of stay after blood cultures were collected, and all-cause in-hospital mortality. Out of a total of 923 patients, 390 (125 baseline, 134 intervention, 131 post-intervention) who were not on optimal therapy at the time of Gram stain or had contaminated blood cultures were assessed. Compared with baseline, only the median time to optimal therapy for MSSA bacteraemia was reduced in both the intervention and post-intervention periods (17 versus 17 versus 50 h; P < 0.001), respectively. In an analysis adjusted for baseline differences among the groups using quantile regression models, use of the Verigene BC-GP assay in both periods significantly reduced time to optimal therapy by 14-22 h in patients who would achieve optimal therapy at ≥ 26 h without the assay. There were no differences in in-hospital mortality or hospital length of stay between study periods. A real-time AS intervention implemented alongside introduction of the Verigene BC-GP assay led to improvements in antibiotic therapy for patients with bacteraemia due to Gram-positive cocci, even after the AS intervention was discontinued.


Dietary restriction alters fine motor function in rats.
A number of standard behavioral tasks in animal research utilize food rewards for positive reinforcement. In order to enhance the motivation to participate in these tasks, animals are usually placed on a restricted diet. While dietary restriction (DR) has been shown to have beneficial effects on recovery after brain injury, life span and aging processes, it might also represent a stressor. Since stress can influence a broad range of behaviors, the purpose of this study was to assess whether DR may have similar effects on skilled movement. Adult male Long-Evans rats were trained and tested in a skilled reaching task both prior to and during a mild food restriction regimen that maintained their body weights at 90-95% of baseline weight for eight days. The observations revealed that DR decreased reaching success and increased the number of attempts to grasp a single food pellet. The animals appeared to be more frantic when attempting to reach for food pellets, and the time taken to reach for 20 pellets decreased following the onset of DR. A second experiment investigating behaviors that do not require food rewards, including a ladder rung walking task and an open field test, confirmed that rats on DR display deficits in skilled movements and are hyperactive. These findings suggest that results obtained in motor tasks using food rewards need to be interpreted with caution. The findings are discussed with respect to stress associated with DR.


Periplasmic expression of Pseudomonas fluorescens peroxidase Dyp1B and site-directed mutant Dyp1B enzymes enhances polymeric lignin degradation activity in Pseudomonas putida KT2440.
Expression of lignin-oxidising Pseudomonas fluorescens Dyp1B in the periplasm of Pseudomonas putida KT2440, using a tat fusion construct, was found to lead to enhanced whole cell activity for oxidation of DCP and polymeric lignin substrates. Four amino acid residues predicted to lie at the manganese ion binding site of Pseudomonas fluorescens peroxidase Dyp1B were investigated using site-directed mutagenesis. Mutants H127R and S223A showed 2-fold and 4-fold higher kcat for Mn(II) oxidation respectively, and mutant S223A showed 2-fold enhanced production of low molecular weight phenolic products from a polymeric soda lignin. The mutant Pfl Dyp1B genes were expressed as tat fusions to investigate their effect on lignin oxidation by P. putida KT2440.


Correlation between Targeted qPCR Assays and Untargeted DNA Shotgun Metagenomic Sequencing for Assessing the Fecal Microbiota in Dogs.
DNA shotgun sequencing is an untargeted approach for identifying changes in relative abundances, while qPCR allows reproducible quantification of specific bacteria. The canine dysbiosis index (DI) assesses the canine fecal microbiota by using a mathematical algorithm based on qPCR results. We evaluated the correlation between qPCR and shotgun sequencing using fecal samples from 296 dogs with different clinical phenotypes. While significant correlations were found between qPCR and sequencing, certain taxa were only detectable by qPCR and not by sequencing. Based on sequencing, less than 2% of bacterial species (17/1190) were consistently present in all healthy dogs (n = 76). Dogs with an abnormal DI had lower alpha-diversity compared to dogs with normal DI. Increases in the DI correctly predicted the gradual shifts in microbiota observed by sequencing: minor changes (R = 0.19, DI < 0 with any targeted taxa outside the reference interval, RI), mild-moderate changes (R = 0.24, 0 < DI < 2), and significant dysbiosis (R = 0.54, 0.73, and 0.91 for DI > 2, DI > 5, and DI > 8, respectively), compared to dogs with a normal DI (DI < 0, all targets within the RI), as higher R-values indicated larger dissimilarities. In conclusion, the qPCR-based DI is an effective indicator of overall microbiota shifts observed by shotgun sequencing in dogs.


Increased Moraxella and Streptococcus species abundance after severe bronchiolitis is associated with recurrent wheezing.
The role of the airway microbiome in the development of recurrent wheezing and asthma remains uncertain, particularly in the high-risk group of infants hospitalized for bronchiolitis. We sought to examine the relation of the nasal microbiota at bronchiolitis-related hospitalization and 3 later points to the risk of recurrent wheezing by age 3 years. In 17 US centers researchers collected clinical data and nasal swabs from infants hospitalized for bronchiolitis. Trained parents collected nasal swabs 3 weeks after hospitalization and, when healthy, during the summer and 1 year after hospitalization. We applied 16S rRNA gene sequencing to all nasal swabs. We used joint modeling to examine the relation of longitudinal nasal microbiota abundances to the risk of recurrent wheezing. Among 842 infants hospitalized for bronchiolitis, there was 88% follow-up at 3 years, and 31% had recurrent wheezing. The median age at enrollment was 3.2 months (interquartile range, 1.7-5.8 months). In joint modeling analyses adjusting for 16 covariates, including viral cause, a 10% increase in relative abundance of Moraxella or Streptococcus species 3 weeks after day 1 of hospitalization was associated with an increased risk of recurrent wheezing (hazard ratio [HR] of 1.38 and 95% high-density interval [HDI] of 1.11-1.85 and HR of 1.76 and 95% HDI of 1.13-3.19, respectively). Increased Streptococcus species abundance the summer after hospitalization was also associated with a greater risk of recurrent wheezing (HR, 1.76; 95% HDI, 1.15-3.27). Enrichment of Moraxella or Streptococcus species after bronchiolitis hospitalization was associated with recurrent wheezing by age 3 years, possibly providing new avenues to ameliorate the long-term respiratory outcomes of infants with severe bronchiolitis.


Utility of cystatin C as a marker for the severity of aortic regurgitation in hypertensive patients.
Cystatin C, a cathepsin inhibitor, is involved in the remodeling of human aortic valve and left ventricle (LV). Cystatin C may be related to the severity of aortic regurgitation (AR). We measured cystatin C and CRP in hypertensive patients with mild-to-moderate AR (n=120) and in those without AR (n=128). Echocardiography was performed to assess the vena contracta width (the narrowest region of regurgitant jet, VCW) as a marker of the severity of AR, relative wall thickness as a marker of LV concentric remodeling, and the ratio of early peak mitral flow to early diastolic mitral annular velocity (E/e') as an index of LV diastolic function. Glomerular filtration rate (GFR) was estimated using the MDRD methods. Cystatin C levels were greater in hypertensive patients with AR than in those without AR. A multiple linear regression analysis indicated cystatin C levels correlated with the VCW independent of GFR, body mass index, CRP, relative wall thickness, and E/e' in hypertensive patients with AR. Cystatin C was associated with the severity of regurgitation independent of renal function body composition chronic inflammation LV remodeling and diastolic function in hypertensive patients with mild-to-moderate AR.


Cutaneous histiocytoses in children.
Cutaneous histiocytoses constitute a heterogeneous group of diseases characterised by the cutaneous accumulation of cells with the cytological and phenotypic features of macrophages or dendritic cells. The clinical spectrum ranges from self-resolving, skin-limited conditions to severe, multiorgan disease with a high morbidity rate. Until recently, cutaneous histiocytoses were classified according to the immunophenotype of the pathological cells, with differentiation between Langerhans cell histiocytosis (LCH) [CD1a+, CD207 (langerin)+] and non-Langerhans cell histiocytosis (CD68+, CD163+, CD1a-, CD207-). Over the last 12 years, a number of new pathophysiological findings (in particular, molecular pathology results) regarding histiocytoses have contributed to a new classification based on molecular alterations, as well as on clinical and imaging characteristics and the phenotype. The most frequent entities in children are juvenile xanthogranuloma and LCH.


A randomized trial of two home-based exercise programmes to improve functional walking post-stroke.
To estimate the relative effectiveness in improving walking ability and other mobility and health outcomes post-stroke of two home-based exercise programmes - stationary cycling and an exercise and walking programme. An observer-blinded, randomized, pragmatic, trial with repeated measures. Hospital centers in two Canadian cities. People within 12 months of acute stroke who were able to walk >10 meters independently and healthy enough to engage in exercise. Two dose-equivalent interventions, one involving stationary cycling and the other disability-targeted interventions were tested. Both protocols required daily moderate intensity exercise at home building up to 30 minutes per day. One group exercised on a stationary bicycle, the second group carried out mobility exercises and brisk walking. The primary outcome was walking capacity as measured by the six-minute walk test (6MWT). Secondary outcomes were physical function, role participation, health-related quality of life exercise adherence, and adverse events. The study failed to meet recruitment targets: 87 participants (cycle group, n = 43; exercise group, n = 44) participated. No significant effects of group or time were revealed for the 6MWT, which was approximately 320 m at randomization. A significant effect for role participation was found in favor of the exercise group (global odds ratio (OR) for cycling vs. exercise was 0.51; 95% confidence interval (CI), 0.27-0.95). Change in the 6MWT between highest and lowest adherence categories was statistically significant (p = 0.022). Both programmes were equally effective in maintaining walking capacity after discharge from stroke rehabilitation; or were equally ineffective in improving walking capacity. Clinical Trials Gov number: NCT00786045.


Cancer risk in inflammatory bowel disease according to patient phenotype and treatment: a Danish population-based cohort study.
Population-based studies of site-specific cancer risk in patients with inflammatory bowel disease (IBD) according to IBD phenotype and treatment are lacking. We studied cancer risk in a well-characterized population-based IBD cohort from North Jutland County, Denmark. A total of 1,515 patients were diagnosed with ulcerative colitis (UC) and 810 with Crohn's disease (CD) during 1978-2002. Patients were followed until 31 December 2010 for occurrence of incident cancer, identified in the Danish Cancer Registry. Observed numbers of cancer were compared with expected numbers (based on age- and sex-specific background rates) and presented as standardized incidence ratios (SIRs) with 95% confidence intervals (CIs). Patients with UC were not at increased risk of cancer overall (SIR, 1.12; 95% CI, 0.97-1.28) despite increased risk of prostate cancer (SIR, 1.82; 95% CI, 1.17-2.71). Patients with CD had a 55% increased risk of cancer overall (SIR, 1.55; 95% CI, 1.29-1.84) related to young age, colonic disease, smoking, and thiopurine exposure. Patients were at increased risk of small bowel cancer (SIR, 15.18; 95% CI, 1.84-54.78), lung cancer (SIR, 2.13; 95% CI, 1.19-3.52 (associated with female gender and smoking)), colorectal cancer in males (SIR, 2.43; 95% CI, 1.05-4.78), cervical dysplasia (SIR, 1.65; 95% CI, 1.10-2.37 (associated with young age at diagnosis, smoking, 5-aminosalicylic acid, and thiopurine exposure)), and non-Hodgkin lymphoma (SIR, 3.43; 95% CI, 1.38-7.07 (unrelated to thiopurine exposure)). Patients with CD, but not UC, have an overall excess risk of cancer. Clinical characteristics of IBD patients at excess risk differ by cancer subtype.


Tricuspid and right ventricular infundibular obstruction associated with calcified right ventricular pseudo-tumour. Radiologic and pathologic aspects.
The authors describe a rare 'pseudo-tumour' in the right heart, discovered after a cerebrovascular accident (paradoxal embolus probably). The mass occupied the anterior leaflet of the tricuspid valve. Its mobility and numerous calcifications facilitated diagnosis, determination of its exact position, and evaluation of haemodynamic consequences. The histopathologic features are reported, and its aetiology is discussed.


A protocol for the treatment of severe infections of the hand.
A protocol for the treatment of hand infections was used for 69 patients in a university teaching hospital. The protocol prescribes incision and drainage under optimal conditions (in the operating room) when the patient is first seen and intravenous antibacterial therapy effective against anaerobic and aerobic bacteria initiated immediately after obtaining aerobic and anaerobic cultures. Results of bacteria studies confirmed the significant frequency (nearly 30%) of mixed aerobic and anaerobic infection. Use of the protocol resulted in a shorter hospital stay, faster healing, and fewer complications (recrudescence of infection, reoperation, stiffness, arthritis, and osteomyelitis) when compared with 107 patients who were treated before institution of the protocol.


Modelling air pollution for epidemiologic research--Part I: A novel approach combining land use regression and air dispersion.
A common limitation of epidemiological studies on health effects of air pollution is the quality of exposure data available for study participants. Exposure data derived from urban monitoring networks is usually not adequately representative of the spatial variation of pollutants, while personal monitoring campaigns are often not feasible, due to time and cost restrictions. Therefore, many studies now rely on empirical modelling techniques, such as land use regression (LUR), to estimate pollution exposure. However, LUR still requires a quantity of specifically measured data to develop a model, which is usually derived from a dedicated monitoring campaign. A dedicated air dispersion modelling exercise is also possible but is similarly resource and data intensive. This study adopted a novel approach to LUR, which utilised existing data from an air dispersion model rather than monitored data. There are several advantages to such an approach such as a larger number of sites to develop the LUR model compared to monitored data. Furthermore, through this approach the LUR model can be adapted to predict temporal variation as well as spatial variation. The aim of this study was to develop two LUR models for an epidemiologic study based in Greater Manchester by using modelled NO(2) and PM(10) concentrations as dependent variables, and traffic intensity, emissions, land use and physical geography as potential predictor variables. The LUR models were validated through a set aside "validation" dataset and data from monitoring stations. The final models for PM(10) and NO(2) comprised nine and eight predictor variables respectively and had determination coefficients (R²) of 0.71 (PM(10): Adj. R²=0.70, F=54.89, p<0.001, NO(2): Adj. R²=0.70, F=62.04, p<0.001). Validation of the models using the validation data and measured data showed that the R² decreases compared to the final models, except for NO(2) validation in the measured data (validation data: PM(10): R²=0.33, NO(2): R²=0.62; measured data: PM(10): R²=0.56, NO(2): R²=0.86). The validation further showed low mean prediction errors and root mean squared errors for both models.


CloneSifter: enrichment of rare clones from heterogeneous cell populations.
Many biological processes, such as cancer metastasis, organismal development, and acquisition of resistance to cytotoxic therapy, rely on the emergence of rare sub-clones from a larger population. Understanding how the genetic and epigenetic features of diverse clones affect clonal fitness provides insight into molecular mechanisms underlying selective processes. While large-scale barcoding with NGS readout has facilitated cellular fitness assessment at the population level, this approach does not support characterization of clones prior to selection. Single-cell genomics methods provide high biological resolution, but are challenging to scale across large populations to probe rare clones and are destructive, limiting further functional analysis of important clones. Here, we develop CloneSifter, a methodology for tracking and enriching rare clones throughout their response to selection. CloneSifter utilizes a CRISPR sgRNA-barcode library that facilitates the isolation of viable cells from specific clones within the barcoded population using a sequence-specific retrieval reporter. We demonstrate that CloneSifter can measure clonal fitness of cancer cell models in vitro and retrieve targeted clones at abundance as low as 1 in 1883 in a heterogeneous cell population. CloneSifter provides a means to track and access specific and rare clones of interest across dynamic changes in population structure to comprehensively explore the basis of these changes.


Protein microarray velcro.
A designed polypeptide scaffold contains a surface immobilization domain and a target capture domain for flexible protein arraying.


Effects of the ruthenium-based drug NAMI-A on the roles played by TGF-β1 in the metastatic process.
The ruthenium-based drug NAMI-A, characterised by its selectivity against solid tumour metastases, promotes TGF-β1-dependent fibrosis and the reduction of the release of MMPs in the primary tumour. The aim of the study was to examine the interaction of NAMI-A with TGF-β1 in the process of metastasis formation. NAMI-A (1) affects the secretion of TGF-β1 in metastatic MDA-MB-231 cells rather than in non-tumorigenic HBL-100 cells, (2) prevails over TGF-β1 with regard to the invasive capacity of the treated cells, and (3) contrasts integrin-dependent migration stimulated by TGF-β1. It, thus, appears that the effects of NAMI-A on cell invasion and migration are best summarised as an interference with TGF-β1 and a reduction of its activity in these events. At a molecular level, the similar activity of NAMI-A and TGF-β1 on RhoA GTPase supports its interaction with cell surface integrins while TGF-β1 can activate it by interaction with its TGFβR receptor. The inhibition of TGF-β1-induced migration of MDA-MB-231 cells by NAMI-A cannot simply be attributed to a modulation of the Smad2 and p38MAPK pathways. In conclusion, the effects of NAMI-A on the biological role of TGF-β1 in cancer metastasis are insufficient to attribute the responsibility for the anti-metastatic activity of the ruthenium-based drug to this target alone.


Sonographic findings in splenogonadal fusion.
Splenogonadal fusion is a rare developmental anomaly usually presenting as a scrotal mass. The imaging findings and etiology of this condition are discussed. Knowledge of this entity may help to prevent unnecessary orchiectomy.


Abortion and methods of reproductive planning: the views of Malta's medical doctor cohort.
Malta is the only EU country where abortion remains illegal in all circumstances. This study aims to assess the previously unexplored views of Malta's medical doctors on the legality of contraception, abortion, assisted reproductive technology and surrogacy. Following ethics approval, 1578 out of a total of 2468 registered medical doctors in Malta were invited for an anonymous survey. The response rate was 28.8% (n = 454), guaranteeing a maximum margin of error of 4.16% assuming a 95% confidence level. Responses consisted of "Yes", "No", and "Not sure". In the abortion section, "Yes" responses were specific to different gestational age limits. A majority supported all contraceptive methods being legal: morning-after pill (59.7%); intra-uterine device (85.9%); surgical sterilisation (>70%). A majority also agreed with in-vitro fertilisation being legal (93.8%). Embryo freezing and surrogacy garnered less support (39.9% and 40.5% respectively). Legalising abortion for "Woman's life in danger" and "Non-viable fetal anomaly" was supported at least up to 12 weeks gestation by 66.8% and 63.0% respectively, and in all stages of pregnancy by 54.2% and 50.2% respectively. Support, at least up to 12 weeks gestation, was less for other circumstances: "Rape or incest" (35.3%); "Preserve a woman's physical health" (30.0%); "Preserve a woman's mental health" (26.8%); "Viable fetal anomaly" (24.6%); "<16 years of age" (23.8%); "Economic/social reasons" (18.9%); "Any circumstance" (14.5%). According to the surveyed respondents, there is at present a clear majority of doctors in Malta who do not agree with the total legal ban on abortion and support its legalisation in limited circumstances.


Clinical and Biomechanical Outcomes of Suture Button Fixation for Ligamentous Lisfranc Injury: A Systematic Review and Meta-analysis.
Flexible ligamentous fixation has increased in popularity for the treatment of ligamentous Lisfranc injury, but the optimal fixation strategy is unclear. To review the biomechanical, clinical, and radiographic results of ligamentous Lisfranc injuries treated with flexible fixation. Systematic review; Level of evidence, 4. A systematic literature review was conducted according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. The PubMed/Medline and Google Scholar literature databases were queried for clinical and biomechanical (cadaveric) studies relating to flexible fixation of ligamentous Lisfranc injury. Outcomes of interest included patient-reported outcome scores, clinical/biomechanical results, radiographic alignment, and return to activity. Where appropriate, meta-analysis of the postoperative outcomes was performed. Of the 34 initial studies, 14 articles (243 feet) were included in the analysis. In the 11 clinical studies (216 patients), the mean postoperative American Orthopaedic Foot & Ankle Society score was 90.1 (n = 150; 6 studies) and the mean visual analog scale score was 1.5 (n = 137; 5 studies). The rate of return to activity was 100% (n = 35; 5 studies), and 100% of patients maintained radiographic alignment postoperatively (n = 62; 6 studies). No complications or subsequent hardware removals were reported. Of the 3 biomechanical studies (27 feet), 1 study found significantly greater change in diastasis under axial load between intact and postfixation ligaments with suture button versus screw fixation (+1.1 vs -0.1 mm; P < .05), another found no difference in the decrease in diastasis under axial load between the injured state and screw or suture button fixation (1.2 vs 1.0 mm; P = .5), and the third found no difference in displacement between intact and either screw or suture button fixation under either axial (intact vs screw: 1.0 vs 2.0 mm, P = .1; intact vs suture button: 0.6 vs 1.8 mm, P = .1) or abduction (intact vs screw: 1.5 vs 1.1 mm, P = .5; intact vs suture button: 1.3 vs 2.1 mm, P = .1) load. Flexible fixation use in the treatment of ligamentous Lisfranc injury was found to have significant potential as a fixation option, as demonstrated by excellent clinical results. Biomechanical evidence was inconclusive but suggested a trend toward decreased diastasis in specimens fixed with screws compared with suture buttons.


S-1 plus irinotecan and oxaliplatin for the first-line treatment of patients with metastatic colorectal cancer: a prospective phase II study and pharmacogenetic analysis.
S-1 is an oral fluoropyrimidine that mimics infusional 5-fluorouracil. The aim of this phase II trial was to explore the clinical efficacy of the triplet regimen TIROX, which consists of S-1, irinotecan and oxaliplatin. Forty-two chemo-naive patients with metastatic colorectal cancer (mCRC) were planned to be enrolled and be treated with irinotecan 150 mg m(-2) followed by oxaliplatin 85 mg m(-2) on day 1 and S-1 80 mg m(-2) per day from day 1 to 14 every 3 weeks. Polymorphisms in the UGT1A1, UGT1A6, UGT1A7 and CYP2A6 genes were analysed. Between July 2007 and February 2008, 43 patients were enrolled. An objective response was noted in 29 patients (67.4%, 95% confidence interval: 53.4-81.4), of which 2 achieved durable complete responses. The median progression-free survival was 10.0 months and the median overall survival was 19.2 months. Significant grade 3 or 4 adverse events were neutropenia (45.2%), febrile neutropenia (9.5%), diarrhoea (7.1%) and vomiting (9.5%). Increased gastrointestinal toxicities were associated with the presence of UGT1A6*2 or UGT1A7*3 and an improved tumour response was noted in those without variant alleles of CYP2A6 or UGT1A1*60. The combination of S-1, irinotecan and oxaliplatin showed favourable efficacy and tolerability in untreated patients with mCRC.


Relationship of sexual dysfunction to epilepsy laterality and reproductive hormone levels in women.
Sexual dysfunction has been reported to be common among women with epilepsy. Controlled studies, quantitative data, and investigations of potentially contributory factors, however, have been few. The purpose of this investigation was to determine if (1) sexual dysfunction is unusually common among women with partial seizures of temporal lobe origin (TLE), and (2) sexual dysfunction varies in relation to the laterality of EEG epileptiform discharges, antiepileptic drug use, and serum gonadal steroid levels. This controlled prospective investigation used a quantitative sexual rating scale and reproductive hormone measures to compare sexual dysfunction in women with left and right unilateral temporolimbic epilepsy and controls. Sexual dysfunction scores were significantly higher in women with TLE, and sexual dysfunction affected substantially more women with epilepsy than controls. Women with right-sided foci were affected more than women with left-sided foci. There was a significant inverse correlation between sexual dysfunction and bioactive testosterone levels in women with epilepsy as well as in controls. Serum estradiol was lower in women with TLE but did not correlate significantly with overall sexual dysfunction. The findings suggest that sexual dysfunction is significantly more common in women with right-sided epileptiform discharges than in controls and is inversely correlated with bioactive testosterone levels. The value of hormonal replacement or supplementation remains to be explored.


Computed tomography for diagnosis of persistent hyperplastic primary vitreous (PHPV).
Leucocoric eyes with persistent hyperplastic primary vitreous (PHPV) were studied by computed tomography (CT). Maximum information was derived from use of an intravenous contrast agent and repetition of the scanning in the lateral decubitus position. Eight diagnostically helpful radiologic findings were noted, some for the first time. (1) Radiodense retrolental soft tissue can be demonstrated along Cloquet's canal. (2) The retrolental tissue enhances after administration of intravenous iodine-containing contrast material. (3) Congenital nonattachment of the retina is demonstrable by CT. (4) There may be localized or generalized increased radiodensity of the vitreous body. However, some minimally affected cases have vitreous chambers with normal attenuation values, and CT evidence for PHPV may be lacking in such patients. (5) There may also be layered, high density fluid (blood) in the retrohyaloid space, which shifts location in the lateral decubitus position. Thus far, this finding appears to be unique to PHPV, but computed scanning of other neonatal ocular conditions will be necessary before concluding it is pathognomonic. (6) There is absence of ocular and intraorbital calcification. (7) Abnormalities in configuration of the eyeball, including microphthalmos, can be demonstrated. (8) Retrobulbar tissues and other orbital structures appear normal, and the optic nerves appear normal or minimally reduced in size.


Stereotactic body radiation therapy for spinal metastases.
The goal of this paper is to expose the clinical results and review of the literature of the treatment of spinal metastases with Stereotactic Body Radiation Therapy (SBRT) presenting one case. A spinal metastases from rectal cancer treated with a single dose of 18Gy is presented. The following physics aspects are exposed: Treatment volume, tumor volume marginal doses and maximum doses in organs at risk. Clinical and radiographic follow up is presented. Local control and pain relief after one year of follow up was excellent. In properly selected patients, the treatment of limited metastatic disease with SBRT appears to be feasible and safe.


Magnetic field reversal in the turbulent environment around a repeating fast radio burst.
Fast radio bursts (FRBs) are brief, intense flashes of radio waves from unidentified extragalactic sources. Polarized FRBs originate in highly magnetized environments. We report observations of the repeating FRB 20190520B spanning 17 months, which show that the FRB's Faraday rotation is highly variable and twice changes sign. The FRB also depolarizes below radio frequencies of about 1 to 3 gigahertz. We interpret these properties as being due to changes in the parallel component of the magnetic field integrated along the line of sight, including reversing direction of the field. This could result from propagation through a turbulent magnetized screen of plasma, located 10-5 to [Formula: see text] parsecs from the FRB source. This is consistent with the bursts passing through the stellar wind of a binary companion of the FRB source.


Cells on lockdown: long-term consequences of CDK4/6 inhibition.
Inhibition of cyclin-dependent kinases Cdk4/6 is emerging as a useful anti-proliferative chemotherapy, but it remains unclear how durable inhibition of cancer cell proliferation is achieved to promote a long-lasting response in patients, or how toxicity is limited to cancer cells with minimal side effects. Two recent papers in The EMBO Journal investigating senescence induction following prolonged Cdk4/6 inhibitor treatment now reveal important insights into ways to increase anti-tumour effects of Cdk4/6 inhibition and to reduce therapy-induced side effects of senescence induction.


Influence of CYP2C9 genotypes on the pharmacokinetics and pharmacodynamics of piroxicam.
Our objective was to evaluate the influence of cytochrome P450 (CYP) 2C9 polymorphisms on the pharmacokinetics and pharmacodynamics of the nonsteroidal anti-inflammatory drug piroxicam. Thirty-five healthy subjects with CYP2C9 genotypes *1/*1 (n=17), *1/*2 (n=9), and *1/*3 (n=9) received a single oral dose of piroxicam (20 mg). Blood samples were collected at various time points up to 240 hours for measurements of the concentrations of piroxicam and thromboxane B2 (TXB2). Piroxicam's area under the plasma concentration-time curve from time 0 to infinity and oral clearance corrected for body weight were 154+/-37 microg.mL-1.h and 2.0+/-0.5 mL.h-1.kg-1, respectively, in CYP2C9*1/*1 individuals, as compared with 256+/-97 mL.h-1 (P=.002) and 1.3+/- 0.4 mL.h-1.kg-1 (P=.002), respectively, in CYP2C9*1/*2 individuals and 259+/- 95 mL.h-1 (P=.002) and 1.3+/- 0.4 mL.h-1.kg-1 (P=.002), respectively, in CYP2C9*1/*3 individuals. There were no significant differences between CYP2C9*1/*2 and CYP2C 9*1/*3 individuals in these pharmacokinetic parameters (P=.95 for area under the plasma concentration-time curve from time 0 to infinity and P=.94 for oral clearance corrected for body weight). The formation of TXB2, reflecting cyclooxygenase type 1 activity, showed significant differences in the area above the effect-time curves (expressed as percent of baseline TXB2.h) between CYP2C9*1/*1 (10,190 +/- 2632) and either CYP2C9*1/*2 (19,255+/-1,291 [P=.00003]) or CYP2C9*1/*3 (18,241+/- 2397 [P=.00003]). The minimum serum TXB2 concentration, however, did not differ among the different genotypes (P=.32, ANOVA). Piroxicam's oral clearance was impaired and its inhibitory effect on cyclooxygenase 1 activity was increased in CYP2C9*1/*2 or CYP2C9*1/*3 individuals, as compared with CYP2C 9*1 homozygous individuals.


Quantifying the future risk of dengue under climate change in Japan.
In metropolitan Tokyo in 2014, Japan experienced its first domestic dengue outbreak since 1945. The objective of the present study was to quantitatively assess the future risk of dengue in Japan using climate change scenarios in a high-resolution geospatial environment by building on a solid theory as a baseline in consideration of future adaptation strategies. Using climate change scenarios of the Model for Interdisciplinary Research on Climate version 6 (MIROC6), representative concentration pathway (RCP) 2.6, 4.5, and 8.5, we computed the daily average temperature and embedded this in the effective reproduction number of dengue, R(T), to calculate the extinction probability and interepidemic period across Japan. In June and October, the R(T) with daily average temperature T, was <1 as in 2022; however, an elevation in temperature increased the number of days with R(T) >1 during these months under RCP8.5. The time period with a risk of dengue transmission gradually extended to late spring (April-May) and autumn (October-November). Under the RCP8.5 scenario in 2100, the possibility of no dengue-free months was revealed in part of southernmost Okinawa Prefecture, and the epidemic risk extended to the entire part of northernmost Hokkaido Prefecture. Each locality in Japan must formulate action plans in response to the presented scenarios. Our geographic analysis can help local governments to develop adaptation policies that include mosquito breeding site elimination, distribution of adulticides and larvicides, and elevated situation awareness to prevent transmission via bites from Aedes vectors.


Exogenous and endogenous orienting of visuospatial attention in P300-guided brain computer interfaces: a pilot study on healthy participants.
Studies on brain computer interfaces (BCIs) have been mainly concerned with algorithm improvement for better signal classification. Fewer studies, however, have addressed to date the role of cognitive mechanisms underlying the elicitation of brain-signals in BCIs. We tested the effect of visuospatial attention orienting on a P300-guided BCI, by comparing the effectiveness of three visual interfaces, which elicited different modalities of visuospatial attention orienting (exogenous vs. endogenous). Twelve healthy participants performed 20 sessions, using the abovementioned P300-guided BCI interfaces to control a cursor. Brain waves were recorded on each trial and were subsequently classified on-line using an ad hoc algorithm. Each time the P300 was correctly classified, the cursor moved towards the target position. The "endogenous" interface was associated with significantly higher performance than the other two interfaces during the testing sessions, but not in the follow-up sessions. Endogenous visuospatial attention orienting can be effectively implemented to increase the performance of P300-guided BCIs. The study of visuospatial attention underlying participants' performance is essential for implementing efficient visual BCIs.


[Etiology of hip dislocation in children with cerebral movement disorders and possibilities of conservative treatment using rotating and spreading plates and electrostimulation].
Dislocations of the hip associated with spastic paralysis are almost always found in severely disabled children. The onset is in infancy. Two methods, electrostimulation of the minor gluteal muscles and treatment with simple leg casts to correct the malposition, are described. Follow-up examinations showed that the treatment had had a positive effect on the hip condition in 33 of the 42 children treated with casts and 26 of the 32 children treated by electrotherapy. Electrotherapy is an innovation, since in the past it was considered contraindicated in these patients. However, it is based on the fact that the gluteal muscles do not normally show any spasticity. Irrespective of this, the treatment is long-term, and has to be continued for many years, a fact which makes greater demands on the cooperativeness of the family.


Bridging the gap between self-assessment and faculty assessment of clinical performance in restorative dentistry: A prospective pilot study.
The current study was undertaken to investigate whether self-assessment of clinical skills of undergraduate dental students could be bridged with faculty assessment by deliberate training over an extended period. A secondary aim was to explore students' perception of self-assessment and its effect on their learning and motivation. MATERIAL AND METHODS: A prospective pilot study was conducted at the Department of Restorative Dentistry at Damascus University. Sixteen students participated in the study, ranging in age between 22 and 23 years. A modified Direct Observation of Procedural Skills form with a grading rubric was used to register and guide students' self-assessment; both were pretested on four students before the study. In total, four clinical encounters were completed by each student. Students were trained on how to conduct proper self-assessment before and after each clinical encounter. A postcourse questionnaire was used to investigate students' perception of self-assessment. Bias in self-assessment decreased consistently after each encounter, and the difference in bias between the first (bias = 0.77) and the last encounter (bias = 0.21) was significant with a medium effect size (p = .022, d = 0.64). The percentage of disconfirming performance dimensions decreased from 39.7% to 26.9%. Students' ability to exactly pinpoint strengths improved consistently and significantly. However, their ability to pinpoint areas of improvement was volatile and showed no significant difference. Bland-Altman graph plots showed higher levels of agreement between self-assessment and faculty assessment. Moreover, students' perception of self-assessment was very positive overall. These findings suggest the possibility that the gap between self-assessment and faculty assessment could be bridged through deliberate training. Future longitudinal research using a larger sample size is still required to further explore whether self-assessment can be actively bridged with faculty assessment by deliberate training.


Rare presentation of double-clonal Waldenström macroglobulinemia with pulmonary embolism: A case report.
Waldenström macroglobulinemia (WM) rarely leads to pulmonary embolism. Due to its low incidence, the underlying pathophysiology, prognosis, and optimal treatment remain largely unexplored and uninvestigated. In this study, a patient with a double-clonal WM, a rare subtype, presented with pulmonary embolism. The patient had a small number of plasma cells without morphological abnormalities, and an effective therapeutic response was observed. Nonetheless, the clinical prognosis requires a long-term follow-up.


An enlarged intramuscular venous malformation in the femoral region successfully treated with complete resection.
Intramuscular venous malformations have been previously described as intramuscular hemangiomas, and various therapies have been applied for their treatment. This condition is relatively rare, and therefore, physicians often struggle to determine the appropriate therapy. We presented a case of an enlarged intramuscular venous malformation relapsed after surgery successfully treated with complete resection. We presented a case of an enlarged intramuscular venous malformation with postoperative recurrence successfully treated with complete resection. A 63-year-old woman presented with a subcutaneous mass in the right distal thigh. She experienced swelling in the right thigh 19 years previously and was diagnosed with a venous aneurysm. Three-dimensional CT angiography confirmed the presence of an irregular vessel assumed to be the feeding vessel, which was dendritically branched from the deep femoral artery. We performed surgical complete resection. Her pain and gait disturbance improved after surgery, and she has not experienced recurrence of the mass for the past 2 years. Conservative therapy is initially used for venous malformations. Sclerotherapy, laser therapy, or surgical resection is considered after low-dose aspirin therapy, in combination with the use of compressive garments. Surgical resection is indicated for completely resectable lesions and is appropriate for large lesions in terms of cosmetic benefit. However, partial resection may result in excessive bleeding or postoperative recurrence. The therapy for venous malformations should be decided based on the degree of disability in daily living, adjacent tissue damage, and cosmetic concerns after appropriate differential diagnostic investigations and biopsy.


Molecular characterization and clinical impact of human bocavirus at a tertiary hospital in Barcelona (Catalonia, Spain) during the 2014-2017 seasons.
The aim was to describe the prevalence, molecular epidemiology and clinical manifestations of human bocavirus (HBoV) in patients attended at a tertiary hospital in Barcelona, Spain. From October 2014 to May 2017, respiratory specimens from paediatric patients were collected for respiratory viruses' laboratory-confirmation. Phylogenetic analyses from partial VP1 sequences were performed from all HBoV laboratory-confirmed specimens. Clinical features were retrospectively studied. 178/10271 cases were HBoV laboratory-confirmed. The median age was 1.53 (IQR 1.0-2.3). Co-detection was highly reported (136; 76%). All viruses belonged into HBoV1 genotype but one into HBoV2. Non-reported mutations were observed and two sites were suggestive to be under negative selection. 61% (109/178) cases had lower RTI (LRTI), of whom 84 had co-detections (77%) and 76 had comorbidities (70%). LRTI was the cause of hospitalization in 85 out of 109 cases (78%), and no differences were found regarding severity factors during hospitalization between co- and single-detections, except for median length of respiratory support, which was longer in cases with co-detections. Close monitoring of predominant HBoV1 showed a high similarity between viruses. The presence of comorbidities might explain the high prevalence of LRTI. Symptomatology in HBoV single-detected cases suggest that HBoV is a true pathogen.


Survival rates and prognostic factors for infiltrating salivary duct carcinoma: Analysis of 228 cases from the Surveillance, Epidemiology, and End Results database.
The survival rates and prognostic factors for salivary duct carcinoma (SDC) are not clear. Survival estimates and prognostic factors were evaluated for 228 patients with SDC identified from the Surveillance, Epidemiology, and End Results (SEER) database. Median overall survival (OS) duration for patients with SDC was 79 months and 5-year disease-specific survival (DSS) rate was 64%. Among patients with SDC with lymph node involvement, larger primary tumor size (>3 cm) was associated with twice the risk of death (p < .03). Factors predictive of improved DSS were age (p = .01), tumor size (p = .006), tumor grade (p = .02), and lymph node involvement (p < .001). Adjuvant radiotherapy did not improve survival when compared to surgery alone for early-stage (I-II) disease (p = .28). Younger patients with SDC (<50 years) showed a better prognosis. Primary tumor size and lymph node involvement were independent and additive risk factors for poor prognosis. The role of adjuvant radiotherapy in the treatment of SDC needs to be explored further.


Characterization and expression profiling of fadd gene in response to exogenous Aeromonas hydrophila or Edwardsiella tarda challenge in the hybrid yellow catfish (Pelteobagrus fulvidraco ♀ × P. vachelli ♂).
In mammals, fas-associated protein with death domain (FADD) is involved in the process of cell apoptosis and plays a key role in innate immune signaling. Nevertheless, its detailed molecular mechanisms underlying apoptosis and immune responses to exogenous bacterial infections in teleosts remain largely unknown. In this study, a group of 60 hybrid yellow catfish (with the body weight of 25 ± 0.5 g) were used in subsequent experiments, we examined the expression profiling of fadd gene through comparative genomics and comparative immunological methods. Our results showed that fadd in the hybrid yellow catfish (hycfadd) exhibited similar gene and spatial structures to those in other vertebrates, and formed an independent clade in phylogeny. An expression pattern analysis revealed that hycfadd widely transcribed in various tissues, with the highest transcription level in the liver. Furthermore, expression profiling of hycfadd when intraperitoneally infected with 50 μL of exogenous Aeromonas hydrophila (2.0 × 107 CFU/mL) or Edwardsiella tarda (2.0 × 107 CFU/mL) within 48 h were significantly up-regulated in the kidney, spleen, liver and intestine. Important genes in the toll like receptor (tlr) 1-tlr2- myeloid differentiation primary response 88 (MyD88)-fadd-caspase (casp) 8 cascades of TLR signaling pathway in liver were significantly up-regulated after the A. hydrophila stimulation, suggesting that apoptosis through the TLR signaling pathway may have been triggered and activated, which were further verified in the liver, kidney, spleen, intestine and gill by a TUNEL assay. Overall, this study provides solid evidence for the bacterial induction of fadd-related apoptosis in teleosts.


[For a better management of the psychic suffering of exiles].
Most people seen by Doctors of the World are foreign nationals exposed to extreme social and administrative precariousness. The violence of their journeys and the reception conditions in France have harmful consequences for their psychological health.


Antibiotic prophylaxis as a quality of care indicator: does it help in the fight against surgical site infections following fragility hip fractures?
Fragility hip fractures are associated with increased morbidity, mortality, and costs. To improve patient care, quality indicator programs were introduced. Yet, the efficacy of these programs and specific quality indicators are questioned. We aimed to determine whether defining prophylactic pre-surgical antibiotic treatment as a quality indicator affected hip fracture outcomes. A retrospective study comparing consecutive patients, 65 years and older, who were operated for fragility hip fractures between 01/01/2011 and 30/06/2016, before and after the prophylactic pre-surgical antibiotic treatment quality indicator, which was introduced in 01/2014. Primary outcomes were 1-year surgical site infections (SSI). Secondary outcomes were meeting the quality index and mortality rates, either within a hospital or during the first post-operative year. 904 patients, ages 82.5 ± 7.2 years were operated for fragility hip fractures. 403 patients presented before the antibiotic prophylaxis quality indicator, and 501 following its administration. Patients demographics were comparable. In the pre-quality indicator period, documentation of prophylactic antibiotic treatment was lacking. Only 19.6% had a record for antibiotic administration in their surgical records and for merely 10.4% the type of antibiotic was stated. However, in the post-quality indicator period, 97.0% of patients had a registered prophylactic antibiotic regimen in the hour preceding the surgical incision (p < 0.001). Post-operative SSI rates were equivalent, and as were in-hospital infections, mortality and recurrent hospitalizations CONCLUSIONS: The introduction of the pre-operative antibiotic treatment quality indicator increased the documentation of antibiotic administration yet failed to influence the incidence of post-operative orthopaedic and medical infections in fragility hip fracture patients.


Effect of high-frequency oscillatory ventilation combined with volume guarantee on preterm infants with hypoxic respiratory failure.
The aim of this study was to assess the effect of volume guarantee (VG) on high-frequency oscillatory ventilation (HFOV) compared with HFOV alone in preterm infants with hypoxic respiratory failure (HRF). Fifty-two preterm infants with HRF refractory to conventional mechanical ventilation (CMV) were enrolled in this study. Between June 2012 and February 2016, HFOV alone was used as rescue therapy when CMV failed for 34 infants, whereas HFOV combined with VG was used as rescue therapy for the other 18 infants between March 2016 and December 2017. HFOV combined with VG resulted in a reduction in the combined outcome of death or bronchopulmonary dysplasia (BPD) (p = 0.017) and also a reduction in episodes of hypercarbia (p = 0.010) compared with HFOV alone. In this study, the preterm infants with HRF ventilated using HFOV combined with VG had a reduced combined outcome of death or BPD and hypercarbia compared with those who received HFOV alone.


A new risk-assessment tool for venous thromboembolism in advanced lung cancer: a prospective, observational study.
Management of cancer-associated venous thromboembolism (VTE) is essential in treatment selection and cancer prognosis. However, to date, there is no method to assess the risk of VTE specifically associated with advanced lung cancer. Our aim was to create a new risk assessment scoring system that can predict the concomitant or incidence of VTE in advanced lung cancer. We used the dataset of 1008 patients with lung cancer in the Rising-VTE/NEJ037 study, of which 100 (9.9%) developed VTE. The items extracted in the multivariate analysis included female sex, adenocarcinoma, performance status, N factor, lymphocyte count, platelet count, prothrombin fragment 1 + 2, and diastolic blood pressure. This model had a maximum score of 8 points, with ≥ 5 points indicating a high risk of VTE. This simple risk-assessment model for VTE complications with advanced lung cancer could help identify cases that required monitoring for VTE.


On the Nonnegative Rank of Euclidean Distance Matrices.
The Euclidean distance matrix for n distinct points in ℝ r is generically of rank r + 2. It is shown in this paper via a geometric argument that its nonnegative rank for the case r = 1 is generically n.


[Experimental study of the anti-inflammatory and analgesic effects of diethyl 1,3-dicyclohexyl- 1,2,3,6-tetrahydropyrimidine-4,5-dicarboxylate].
To study the anti-inflammatory and analgesic activities of diethyl 1,3-dicyclohexyl-1,2,3,6-tetrahydropyrimidine-4,5-dicarboxylate (ZL-5010) in vivo and in vitro. The analgesic effect of ZL-5010 was evaluated by acetic acid-induced writhing response in mice, and the anti-inflammatory effects was assessed in mice with xylene-induced ear edema and in rats with carrageenan-induced paw edema. Mouse peritoneal exudate cells activated by bacterial lipopolysaccharides (LPS) were used to evaluate the anti-inflammatory effect of ZL-5010 in vitro. The levels of interleukin-1β (IL -1β) and tumor necrosis factor-α (TNF-α) in the cell culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA). At the doses of 0.25 and 0.5 mmol/kg, ZL-5010 administered by gavage once daily for 3 days significantly reduced acetic acid-induced writhing frequency and suppressed xylene-induced ear edema in mice, and alleviated paw edema induced by carrageenan in rats (P<0.05). The agent also inhibited the production of the pro-inflammatory cytokines IL-1β and TNF-α by LPS-induced mouse peritoneal exudate cells in vitro, with the statistically significant minimum effective concentrations of 10 and 20 µmol/L, respectively (P<0.05). ZL-5010 administered by gavage has anti-inflammatory and analgesic effects in mice and rats, and in mouse peritoneal exudate cell cultures, the agent also inhibits the production of the pro-inflammatory cytokines IL-1β and TNF-α.


Sudden total bilateral deafness due to asymptomatic mumps infection.
Mumps is the most common cause of unilateral acquired sensorineural hearing loss in children. Although it usually affects the salivary glands. the inner ear may be involved. Deafness is usually unilateral, sudden in onset, profound and permanent. Bilateral total sensorineural hearing loss had been rarely reported in English literature. We present a case of total deafness due to asymptomatic mumps infection.


Inhibitory and excitatory binocular convergence to visual cortical neurons of the cat.
Responses of 182 visual cortical (VC) neurons to electrical stimulation of both optic nerves (ON) were recorded intra- and extracellularly, and their eye dominance determined with visual stimuli. Many VC neurons could not be excited by ON stimulation without simultaneous activation by visual stimulation of the eye of the other side. The ON-excited units (25 simple and 71 complex cells) had essentially the same response latency from both ONs. In almost all VC neurons including those not driven from the ON, ON stimulation elicited inhibition which was shown to be post-synaptic in all neurons recorded intra- or quasi-intracellularly. The onset latency of IPSPs not preceded by an action potential or EPSP was 4.2+/-1.0 msec, suggesting that intracortical inhibition was initiated by afferent impulses mediated through fast conducting fibers. In visually monocular neurons, ON stimulation of the non-driving side also elicited primary inhibition. Visually binocular, but monocular-dominant neurons responded more reliably to ON stimulation of the dominant side than to the other. In most binocular neurons with equal visual responsiveness to both eyes, inhibition and excitation evoked from both ONs had about the same latency and magnitude.


Morphometry and morphology of nucleus of the Sertoli and interstitial cells of the tambaqui Colossoma macropomum (Cuvier, 1881) (Pisces: Characidae) during the reproductive cycle.
This study allowed the characterization of the tambaqui Colossoma macropomum testes structural organization, emphasizing Sertoli and interstitial cells and analyzing morphometrically the Sertoli cell nucleus diameter and the interstitial tissue area during the reproductive cycle. Fragments of tambaqui testes were collected in the following reproductive cycle stages: immature, resting, maturation I and II, mature, and regression, and were histologically processed. The Sertoli cells were found at the periphery of the cysts of germinative lineage cells and the nuclei were shown to be smaller as these cells developed. The interstitial cells were better observed between the seminiferous lobules next to vessels in the interstitial tissue of maturing testes.


Novel TetR family transcriptional factor regulates expression of multiple transport-related genes and affects rifampicin resistance in Mycobacterium smegmatis.
Transport-related genes significantly affect bacterial antibiotic resistance. However, the effects of these genes and their regulation of bacterial drug resistance in several mycobacterial species, including the fast-growing Mycobacterium smegmatis, the pathogen M. tuberculosis and M. avium have not been clearly characterized. We identified Ms4022 (MSMEG_4022) as a novel TetR family regulator that activates the expression of seven transport-related genes and affects drug resistance in M. smegmatis. Overexpression of Ms4022 inhibited M. smegmatis growth and enhanced mycobacterial resistance to the anti-tuberculosis drug rifampicin (RIF). By contrast, the Ms4022-deleted mycobacterial strain has shown sensitive to RIF. Ms4022 recognized three 19 bp non-palindromic motifs containing a 9 bp conserved region at their 5' end and it directly regulated seven transport-related genes, which affects mycobacterial resistance to RIF. Overexpression of three of seven transport-related genes (Ms1448, Ms1613, and Ms5278) inhibited the growth of M. smegmatis. This study improves our understanding of the function of mycobacterial transport-related genes and their regulation of bacterial drug resistance.


Romantic relationship status biases memory of faces of attractive opposite-sex others: evidence from a reverse-correlation paradigm.
Previous research has demonstrated that, presumably as a way to protect one's current romantic relationship, individuals involved in a heterosexual romantic relationship tend to give lower attractiveness ratings to attractive opposite-sex others as compared to uninvolved individuals (i.e., the derogation effect). The present study importantly extends this research by examining whether romantic relationship status actually biases memory for the facial appearance of attractive (vs. unattractive) mates. To address this issue, we used a reverse-correlation technique (Mangini & Biederman, 2004), originally developed to get a visual approximation of an individual's internal representation of a target category or person. In line with the derogation effect, results demonstrated that romantically involved (vs. uninvolved) individuals indeed held a less attractive memory of a previously encountered attractive mate's face. Interestingly, they also held a more attractive memory of an unattractive mate's face as compared to uninvolved individuals. This latter finding may suggest that romantically involved (as compared to uninvolved) individuals differentiate opposite-sex others along the attractiveness dimension less.


Are asthma-like symptoms due to bronchial or extrathoracic airway dysfunction?
Patients with asthma-like symptoms may not have asthma but obstruction of the extrathoracic airway (EA). To evaluate if dysfunction of the EA causes asthma-like symptoms, we assessed bronchial and EA responsiveness to inhaled histamine in 441 patients who presented with at least one of three key symptoms--cough, wheeze, dyspnoea--but had neither documented asthma nor bronchial obstruction. The histamine concentrations causing a 20% fall in forced expiratory volume in 1 s (PC20FEV1) and a 25% fall in maximal mid-inspiratory flow (PC25MIF50) were used as respective thresholds of bronchial and EA responsiveness. Values 8 mg/mL or less indicated bronchial (B-HR) or EA hyper-responsiveness (EA-HR). The influence of concurrent upper respiratory tract diseases, such as post-nasal drip (PND), pharyngitis, laryngitis and sinusitis, was also assessed. We found four response patterns to the histamine challenge: EA-HR in 26.5% of the patients, B-HR in 11.1%, combined EA-HR and B-HR in 40.6%, and no-HR in 21.8%. Cough was reported by 79% of the patients, wheeze by 53%, and dyspnoea by 40%. Patients with cough as the sole presenting symptom (34.2%), as compared with those with wheeze and/or dyspnoea (20%), had significantly greater probability of having EA-HR (OR 5.35, 95% CI 3.25-8.82) and lower probability of having B-HR (OR 0.45, CI 0.28-0.70); patients with cough plus wheeze and/or dyspnoea (45.8%) had significantly greater probability of having both EA-HR and B-HR than either those with cough alone (OR 2.48, CI 1.49-4.13), or those with wheeze and/or dyspnoea but not cough (OR 1.74, CI 1.36-2.22). EA-HR alone or combined with B-HR was strongly associated with EA diseases, particularly pharyngitis and PND. Cough was significantly associated with PND, either when it was the sole symptom (OR 2.16, CI 1.14-4.09) or when it was combined with wheeze and/or dyspnoea (OR 3.53, CI 1.97-6.33). Our results suggest that extrathoracic airway dysfunction may account for asthma-like symptoms, particularly chronic cough. This abnormality seems to be sustained by chronic diseases of the upper respiratory tract.


C(alkyl)-C(vinyl) bond cleavage enabled by Retro-Pallada-Diels-Alder reaction.
Activation and cleavage of carbon-carbon (C-C) bonds is a fundamental transformation in organic chemistry while inert C-C bonds cleavage remains a long-standing challenge. Retro-Diels-Alder (retro-DA) reaction is a well-known and important tool for C-C bonds cleavage but less been explored in methodology by contrast to other strategies. Herein, we report a selective C(alkyl)-C(vinyl) bond cleavage strategy realized through the transient directing group mediated retro-Diels-Alder reaction of a six-membered palladacycle, which is obtained from an in situ generated hydrazone and palladium hydride species. This unprecedented strategy exhibits good tolerances and thus offers new opportunities for late-stage modifications of complex molecules. DFT calculations revealed that an intriguing retro-Pd(IV)-Diels-Alder process is possibly involved in the catalytic cycle, thus bridging both Retro-Diels-Alder reaction and C-C bond cleavage. We anticipate that this strategy should prove instrumental for potential applications to achieve the modification of functional organic skeletons in synthetic chemistry and other fields involving in molecular editing.


Cephalic venous flap: a series of 8 cases and a preliminary report on the use of 99mTc labelled RBCs to study the saphenous venous flap in dogs.
From December 1985 to December 1987 a total of eight fasciocutaneous island flaps with only a proximal venous pedicle were performed. Seven of these eight survived completely. The standard flap design and operative technique are given. The results are presented in a tabulated form. One case is presented with illustrations. A preliminary report on the appearance of 99mTc tagged RBCs and Evans blue in a proximal venous saphenous island flap in the dog is included. Comments on the survival of these flaps and the solitary case where the flap necrosed completely are added.


The expression of leptin receptor in the ovary of the queen: leptin receptor expression in queen ovary.
Leptin is a Ob gene product secreted mainly by adipose tissue. Several reports showed leptin production by other tissue including the ovary. The action of leptin is mediated upon binding to its receptor widely expressed in reproductive tissues in different species. In fact, there are growing evidences that leptin plays an important role in the modulation of reproductive functions. Therefore, the aim of this study was to evaluate in the queen, the expression of leptin receptor during the functional ovarian cycle and pregnancy. We found that the ovaries of the queen express leptin receptor in all the examined phases. The highest leptin receptor expression was found in the luteal phase (pseudopregnancy, pregnancy) compared to other phases of the cycle (anestrus, proestrus, estrus). The variations in the expression of leptin receptor suggest a likely implication of leptin in the modulation of ovarian activity, in the examined species.


Effect of desloratadine therapy on symptom scores and measures of nasal patency in seasonal allergic rhinitis: results of a single-center, placebo-controlled trial.
Desloratadine reduces symptoms and maintains nasal airflow in patients with seasonal allergic rhinitis (SAR) during experimental allergen exposure. To compare the effects of desloratadine and placebo on symptom scores, quality of life (QOL), and nasal airway patency in patients with SAR during the allergy season. Adults with symptomatic SAR were randomized in a double-blind manner to receive desloratadine, 5 mg, or placebo for 14 days. Patient-rated SAR symptoms were recorded twice daily (morning and evening). On days 1 and 15, SAR symptoms were scored jointly (investigator and patient), nasal airflow was measured using 4-phase rhinomanometry, and QOL and the overall condition of SAR were rated. Overall treatment response was scored on day 15. Adverse events were recorded. At day 15, total symptom (P = .03) and total nasal symptom (P = .02) scores and patient morning-rated individual nasal symptom scores (except nasal stuffiness) (P < or = .04) decreased significantly from baseline with desloratadine vs placebo. Flow in the descending expiratory nasal airflow phase was significantly greater (P = .046) and the percentage increase in total inspiratory nasal airway resistance was less (P = .03) in the desloratadine group vs the placebo group. The overall condition of SAR was less severe (P = .045), the therapeutic response was greater (P = .004), and the nasal symptom domain of the QOL score was significantly better (P = .03) in the desloratadine group. Adverse event rates were similar in both groups. Desloratadine treatment for 14 days improved nasal airflow and resistance as well as symptom and QOL scores in patients with symptomatic SAR during the allergy season.


Infant speech perception bootstraps word learning.
By their first birthday, infants can understand many spoken words. Research in cognitive development has long focused on the conceptual changes that accompany word learning, but learning new words also entails perceptual sophistication. Several developmental steps are required as infants learn to segment, identify and represent the phonetic forms of spoken words, and map those word forms to different concepts. We review recent research on how infants' perceptual systems unfold in the service of word learning, from initial sensitivity for speech to the learning of language-specific sound patterns. Building on a recent theoretical framework and emerging new methodologies, we show how speech perception is crucial for word learning, and suggest that it bootstraps the development of a separate but parallel phonological system that links sound to meaning.


Low-level laser irradiation affects the release of basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), and receptor of IGF-I (IGFBP3) from osteoblasts.
It was the aim of the present study to evaluate whether the laser irradiation of osteoblasts could enhance the release of growth factors including basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), and receptor of IGF-I (IGFBP3). Low-level laser therapy (LLLT) has been shown to have biostimulatory effects on various cell types by enhancing production of some cytokines and growth factors. Human mesenchymal stem cells (MSCs) were seeded in osteogenic medium and differentiated into osteoblasts. Three groups were formed: in the first group (single dose group), osteoblasts were irradiated with laser (685 nm, 25 mW, 14.3 mW/cm(2), 140 sec, 2 J/cm(2)) for one time; and in the second group, energy at the same dose was applied for 2 consecutive days (double dose group). The third group was not irradiated with laser and served as the control group. Proliferation, viability, bFGF, IGF-I, and IGFBP3 levels were compared between groups. Both of the irradiated groups revealed higher proliferation, viability, bFGF, IGF-I, and IGFBP3 expressions than did the nonirradiated control group. There was increase in bFGF and IGF-I expressions and decrease in IGFBP3 in the double dose group compared to single dose group. The results of the present study indicate that LLLT increases the proliferation of osteoblast cells and stimulates the release of bFGF, IGF-I, and IGFBP3 from these cells. The biostimulatory effect of LLLT may be related to the enhanced production of the growth factors.


Nanofocusing of X-ray free-electron lasers by grazing-incidence reflective optics.
Total-reflection mirror devices for X-ray free-electron laser focusing are discussed in terms of optical design, mirror-fabrication technology, a wavefront diagnosis method and radiation-damage testing, as a review of the present status of the focusing optics at the SPring-8 angstrom compact free-electron laser (SACLA). Designed beam sizes of 1 µm and 50 nm, and spot sizes almost matching prediction have been achieved and used to explore topics at the forefront of natural science. The feasibility of these devices is determined to be sufficient for long-term and stable operation at SACLA by investigating the radiation-damage threshold and achievable accuracies in the mirror figure and alignment.


Gender-based salary differences in academic medicine: a retrospective review of data from six public medical centers in the Western USA.
We assessed the effect of gender, rank and research productivity on compensation for faculty at academic medical centres. A web-based retrospective review of salary for professors in 2016. Faculty from six state-run, publicly funded academic medical centres in the Western USA. 799 faculty members, 225 assistant (51% women), 200 associate (40% women) and 374 full professors (32% women) from general surgery (26% women), obstetrics and gynaecology (70% women) and radiology (34% women). Archived online faculty profiles were reviewed for gender, rank and compensation (total, baseline and supplemental). Total compensation was defined as baseline compensation plus supplemental income. Baseline compensation was defined as base salary minus reductions due to participation in the voluntary Employee Reduction in Time and phased retirement programmes. Supplemental income was defined as additional salary for clinical care and research (eg, grants). Elsevier's Scopus was used to collect data on h-index, a measure of research productivity. Linear regression models were estimated to determine the relationship between these factors and salary. Total compensation was significantly higher for men across all professorial ranks in both general surgery [Formula: see text] and obstetrics and gynaecology [Formula: see text]. Women faculty members within these departments earned almost US$75 000 less than their men colleagues. The disparity in salary originates from gaps in supplemental income, as baseline compensation was not significantly different between men and women. No significant gender difference in total compensation for radiology was found [Formula: see text]. Higher h-index was associated with higher baseline compensation across all departments as well as with supplemental income for general surgery. Higher h-index was related to lower supplemental income for radiology and was not related to supplemental income for obstetrics and gynaecology. Further investigations should focus on discrepancies in supplemental income, which may preferentially benefit men.


Trends in and outcomes from sentinel lymph node biopsy (SLNB) alone vs. SLNB with axillary lymph node dissection for node-positive breast cancer patients: experience from the SEER database.
Complete axillary lymph node dissection (ALND) after a positive sentinel lymph node biopsy (SLNB) remains the standard practice. As nodal surgery has long been considered a staging procedure without a clear survival benefit, the need for ALND in all patients is debatable. The purpose of this study was to examine differences in survival for patients undergoing SLNB alone versus SLNB with complete ALND. Patients with breast cancer who underwent SLNB and were found to have nodal metastases were identified from the Surveillance, Epidemiology, and End Results database (1998-2004). Clinicopathologic and outcomes data were examined for patients who underwent SLNB alone versus SLNB with ALND. We identified 26,986 patients with disease-positive lymph nodes; 4,425 (16.4%) underwent SLNB alone, and 22,561 (83.6%) underwent SLNB with ALND. Patients were significantly more likely to undergo SLNB alone if they were older (median 59 years old) or if the tumor was low grade and estrogen receptor positive. From 1998 to 2004, the proportion of patients with micrometastasis in the sentinel lymph nodes who underwent SLNB alone increased from 21.0 to 37.8% (P < 0.001). At a median follow-up of 50 months, there were no statistically significant differences in overall survival (OS) between patients who underwent SLNB alone versus complete ALND. There is an increasing trend toward omitting ALND in patients with micrometastatic nodal disease identified by SLNB. Compared with SLNB alone, completion ALND does not seem to be associated with improved survival for breast cancer patients with micrometastasis in the sentinel lymph nodes.


Pharmacokinetic study of intravenously administered artemisinin-loaded surface-decorated amphiphilic γ-cyclodextrin nanoparticles.
Artemisinin and its derivatives are currently recommended by World Health Organization for the treatment of malaria. Severe malaria requires a parenteral administration of artemisinin-based formulations. However, the effective use of artemisinin is limited by the pharmacokinetic characteristics of the drug (low water solubility, poor bioavailability and short half-life). To overcome some of these drawbacks, artemisinin-loaded surface-decorated nanoparticles were prepared by co-nanoprecipitation of γ-cyclodextrin bioesterified with C10 alkyl chains and polyethylene glycol (PEG) derivatives (polysorbate 80 and DMPE-mPEG2000). Using a single dose (1.5 mg kg-1 or 2 mg kg-1) by intravenous administration, we investigated the in vivo pharmacokinetic properties in healthy rats of two types of artemisinin-loaded nanoparticle formulations, namely, nanosphere and nanoreservoir systems versus an ethanolic-aqueous solution of artemisinin as reference. Significantly enhanced pharmacokinetic parameters were obtained with artemisinin-loaded nanoparticles. In comparison to reference formulation, the geometric mean exposures in plasma (AUC0-t) exhibited 2.35 and 3.26-fold increases when artemisinin was loaded in nanoreservoir and nanosphere systems, respectively. Its plasma half-life increased 4.00 and 6.25-fold and its clearance decreased up to 2.5 and 4.72-fold. Artemisinin was successfully administered intravenously by means of surface-decorated amphiphilic γ-cyclodextrin nanostructures and showed a longer elimination half-life with respect to an artemisinin solution in ethanol. Therefore, these systems are likely to provide significant advantages for the intravenous treatment of severe malaria.


Synthesis and evaluation of cholesterol-grafted PEGylated peptides with pH-triggered property as novel drug carriers for cancer chemotherapy.
Multifunctional core/shell micelles were self-assembled from triblock copolymers poly(ethylene glycol) methyl ether-b-peptide-g-cholesterol (mPEG-b-P-g-Chol) and used as the doxorubicin delivery carriers for cancer chemotherapy. The copolymers were designed and synthesized successfully based on peptides containing histidine residues (pH-trigger) with different topological structures. The peptides were modified by mPEG (hydrophilic) and cholesterol motifs (hydrophobic) on the terminus, resulting in pH-sensitive amphiphilic copolymers. The critical micelle concentrations (CMCs) of the micelles were determined as 4.79, 2.50 and 1.86mg/L for the linear, Y-shape and fork-shape copolymers, respectively, demonstrating the formation of micelle even at low concentration. The pKb values of three copolymers were found to be around 6.1-6.3 by potentiometric titration test, showing the satisfied pH-sensitivity. The average diameter and zeta potential of blank micelles were 170nm and +20mV at pH 7.4, and increased to 250nm and +35mV at pH 5.0. DOX was loaded into the core of polymeric micelles by dialysis method, and the drug loading capacity slightly increased when the copolymer topological structure changed from linear to Y- and fork-shape. The drug release rate from the system was obviously influencing by the pH values according to the results of in vitro DOX release experiment. Moreover, to investigate the structure-property relationship, the drug release mechanism was preliminarily explored by the semi-empirical equations. Toxicity test showed that three copolymers had bare toxicity whereas the DOX-loaded micelles remained high cytotoxicity for tumor cells. The results indicate the synthesized copolymers might be a potential hydrophobic drug delivery carrier for cancer targeting therapy with controlled drug release.


Responses of goat ruminal musculature to substance P in vitro and in vivo.
A concentration-response curve obtained for contraction of goat rumen strips indicated that substance P (SP) is a very potent spasmogenic agent. In vivo, SP caused a dose-dependent inhibition of the extrinsic rumen contractions possibly due to an increase in ruminal smooth muscle tone, resulting in reflex inhibition. SP did not induce significant changes in heart rate. Further studies are necessary to elucidate a possible physiological role of SP in the modulation of forestomach motility.


Accelerated junctional rhythm (AJR) revealing light-chain cardiac amyloidosis: A case report with literature review.
Cardiac amyloidosis (CA) is a rare condition, characterized by fibrillary proteins infiltration in the extracellular space of the heart. Even though many types of cardiac amyloidosis exist, light chain amyloidosis (AL) and transthyretin amyloidosis (ATTR) remain the most described forms. The diagnosis of amyloidosis represents a real challenge for clinicians, requiring both invasive and non-invasive investigations. Conduction defects and atrial arrhythmias are well known complications of cardiac amyloidosis. However, only a few studies have reported junctional rhythm a primary presentation of light chain cardiac amyloidosis (AL). An early diagnosis and proper management are crucial to improve the prognosis of this disease. Here, we report a rare case of a 48 year-old patient, in acutely decompensated heart failure, presenting an accelerated junctional rhythm (AJR) as initial presentation of light-chain cardiac amyloidosis. The diagnosis was made based on clinical, biological, radiological and histological findings. This case shows diagnostic difficulties and management of this rare disease.


Genetic characterization of dengue virus from patients presenting multi-serotypic infections in the Northern West Bengal, India.
Northern West Bengal, popularly known as North Bengal, is a dengue-endemic area, which has been severely affected by Dengue in the past few years resulting in massive hospitalizations and deaths. Genetic characterization of the circulating endemic dengue virus (DENV) serotypes is of paramount importance for the epidemiological understanding of the infection and subsequent vaccine development. The present study was conceived to characterize circulating dengue serotypes and to undertake phylogenetic study. EDTA blood samples of all (N = 83) NS1-positive cases of patients with acute febrile illness referred to different health care facilities were collected and processed for RNA isolation followed by the complementary DNA (cDNA) preparation. Serotype determination of dengue infection was done using conventional PCR by targeting the viral C-prM region. Phylogenetic tree was constructed by implementing the Maximum likelihood method. Out of 83 blood samples 17 were detected to be positive for the presence of dengue viral RNA. DENV3 was found to be the predominant serotype in the single-infection cases; however, we have detected multi-serotypic co-infections throughout the study. Joint pain was found to be the most valuable symptom for the prognosis of dengue. Sequence analyses suggested that both DENV1- and DENV3-circulating genotypes are in the genotype III group and remain closely related to the Indian clade. To the best of our knowledge, this is the first report on the genetic characterization of circulating DENVs in North Bengal, which may contribute to the study of dengue epidemic and pathogenesis.


Surgical Technique: Treatment of Distal Humerus Nonunions.
Open reduction and internal fixation of distal humerus fractures is standard of care with good to excellent outcome for most patients. However, nonunions of the distal humerus still occur. These are severely disabling problems for the patient and a challenge for the treating physician. Fortunately, a combination of standard nonunion techniques with new plate designs and fixation methods allow even the most challenging distal humeral nonunion to be treated successfully. The purpose of this manuscript is to describe our current technique in treating distal humeral nonunion as it has evolved over the last four decades. We have now follow-up on 62 treated patients. A few key steps are essential to obtain bone healing while regaining or preserving elbow motion. These include careful planning, extensile exposure, release of the ulnar nerve, capsular release and mobilization of the distal fragment, debridement, and finally stable fixation after alignment with application of bone graft. The vast majority of distal humeral nonunions can be treated successfully with open reduction and internal fixation. Important components of the treatment plan are careful preoperative planning, extensile approach, debridement, and solid fixation with-locking-plates and liberal use of bone graft.


Neuroendocrine-immune interactions in synovitis.
Synovial tissue lines the noncartilaginous surfaces of synovial joints and supplies these avascular structures with nutrients. In diseases such as rheumatoid arthritis, inflammation of the synovial tissue--synovitis--induces diffuse damage to the joints. The presence of functional receptors for glucocorticoids, androgens and estrogens in synoviocytes might link inflammation and the endocrine system at the local level. Synovial tissue could be regarded as an intracrine tissue, whereby active steroids influence the cells in which they are synthesized, without their release into the extracellular space. An increase in the peripheral metabolism of sex steroids is characteristic of rheumatoid synovitis, with an augmented ratio of estrogen to androgen occurring in both male and female patients. Changes in the peripheral nervous system at the site of local inflammation are also hallmarks of synovitis in rheumatoid arthritis. In the chronic phase of synovitis, sympathetic nerve fibers are lost; by contrast, sensory nerve fibers sprout into the inflamed tissue. Complex interactions occur between the endocrine, nervous and immune systems during synovitis. In particular, studying neuroendocrine-immune interactions in the inflamed synovium will potentially uncover new mechanisms in the pathophysiology of rheumatoid arthritis and might lead to new methods of therapeutic intervention.


Suppressive effects of a selective inducible nitric oxide synthase (iNOS) inhibitor on pancreatic beta-cell dysfunction.
Type 1 diabetes mellitus is an autoimmune disease characterized by dysfunction and destruction of the pancreatic beta cells. Interleukin-1beta (IL-1beta) has been reported to cause suppression of insulin secretion from pancreatic islets via induction of inducible nitric oxide synthase (iNOS) followed by nitric oxide (NO) production. In this study, we investigated the effects of inhibition of iNOS on pancreatic beta-cell dysfunction in non-obese diabetic (NOD) mice and IL-1beta-treated isolated rat pancreatic islets using a novel specific inhibitor, ONO-1714. Female NOD mice which received subcutaneous infusion of ONO-1714 (4 microg/kg/day or 40 microg/kg/day) from 10 to 14 weeks after birth were compared with untreated NOD mice. In addition, pancreatic islets were isolated from Sprague-Dawley rats and cultured for 24 h with IL-1beta (100 U/mL) with or without ONO-1714 or the non-selective NOS inhibitor NG-monomethyl-L-arginine (L-NMMA). We measured insulin secretion and insulin content of the islets by ELISA, iNOS mRNA expression by reverse transcription-polymerase chain reaction, and NO generation by Griess Reagent System. Hyperglycaemia was observed in NOD mice. ONO-1714 treatment blunted this increase and tended to preserve insulin secretion, although body weight increase did not differ between the groups. Insulitis was also attenuated in the ONO-1714-administered group compared to the control group. Furthermore, in isolated rat pancreatic islets ONO-1714 prevented IL-1beta-induced inhibition of insulin secretion, this protection being evident in much lower concentrations than with L-NMMA. While ONO-1714 completely inhibited IL-1beta-induced NO production, it did not reduce expression of islet iNOS mRNA. These findings indicate that ONO-1714 is promising as a therapeutic agent for autoimmune diabetes.


Modular Peptide Probe for Protein Analysis.
The analysis and regulation of proteins are of great significance for the development of disease diagnosis and treatment. However, complicated analytical environment and complex protein structure severely limit the accuracy of their analysis results. Nowadays, ascribing to the editability and bioactivity of peptides, peptide-based probes could meet the requirements of good selectivity and high affinity to overcome the challenges. In this review, we summarize the advances in the use of modular peptide probes for proteins analysis. It focuses on how to design and optimize the structure of probes, as well as their performance. Then, the strategies and application to improve the analysis result of modular peptide probes are introduced. Finally, we also discuss current challenge and provide some ideas for the future direction for modular peptide probes, hoping to accelerate their clinical transformation.


Climatic proteoglycan stromal keratopathy, a new corneal degeneration.
To characterize a previously undescribed corneal disorder, climatic proteoglycan stromal keratopathy. We prospectively characterized the disorder in 58 eyes of 29 patients (ages 41 to 84 years) at the El Maghraby Eye Hospital, Jeddah Saudi Arabia. Light and transmission electron microscopy were performed on six corneal specimens. Clinical criteria for the diagnosis included a central, horizontally oval, corneal stromal haze with a ground glass appearance, a uniform or lamellar pattern, greater extent anteriorly and involvement of 50% to 100% of stromal thickness. It occurred bilaterally but sometimes asymmetrically. Density varied from mild to moderate and seldom decreased visual acuity. Sixteen (28%) of 58 corneas had additional degenerative lesions (spheroidal degeneration, mild calcific band keratopathy, and Salzmann's nodular degeneration). Refractile stromal lines or posterior white dots were present in six (10%) of 58 eyes. Videokeratography in 11 (29%) of 38 eyes demonstrated flat areas that overlay the stromal haze. Pathologic analysis demonstrated focal intracellular and extracellular deposits of excess proteoglycans. One specimen showed amyloid. We postulate that climatic factors play a pathogenic role because the disorder occurs in individuals who were exposed to the sunny, dry, dusty environment of the Middle East and because other corneal degenerative disorders were present concomitantly. Climatic proteoglycan stromal keratopathy is a corneal degeneration that appears as an oval, central, gray, ground-glass haze associated with central corneal flattening on videokeratography and as excess proteoglycans on histologic examination. It seldom severely decreases visual function.


The history of mutational pressure changes during the evolution of adeno-associated viruses: A message to gene therapy and DNA-vaccine vectors designers.
The use of virus-associated vectors for gene therapy and vaccination have emerged as safe and effective delivery system. Like all other genetic materials, these vehicles are also prone to spontaneous mutations. To understand what types of nucleotide mutations are expected in the vector, one needs to know distinct characteristics of mutational process in the corresponding virus. In this study we analyzed mutational pressure directions along the length of the genomes of all types of primate adeno-associated viruses (AAV) that are frequently used in gene therapy or DNA-vaccines. We observed clear evidences of transcription-associated mutational pressure in AAV: nucleotide usage biases are changing drastically after each of the three promoters: the higher the rate of transcription, the stronger the bias towards GC to AT mutations. Moreover, the usage of G decreased at the lower transcription rate (after P19 promoter) than the usage of C (after P40 promoter). Since nucleotide usage biases are retrospective indices, we created a scenario of changes in transcriptional map during the AAV evolution. Current mutational pressure directions are different for AAV types, while all of them demonstrate high rates of T to C transitions in the second long ORF. Since transcription rate and cell tropism are the main factors determining the preferable direction of nucleotide mutations in AAV, mutational pressure should be checked experimentally in DNA vectors before their final design with the aim to make the transferred gene more stable against those mutations.


Specific sphingolipid content decrease in Cerkl knockdown mouse retinas.
Sphingolipids (SPLs) are finely tuned structural compounds and bioactive molecules involved in membrane fluidity and cellular homeostasis. The core sphingolipid, ceramide (CER), and its derivatives, regulate several crucial processes in neuronal cells, among them cell differentiation, cell-cell interactions, membrane conductance, synaptic transmission, and apoptosis. Mutations in Ceramide Kinase-Like (CERKL) cause autosomal recessive Retinitis Pigmentosa and Cone Rod Dystrophy. The presence of a conserved lipid kinase domain and the overall similarity with CERK suggested that CERKL might play a role in the SPL metabolism as a CER kinase. Unfortunately, CERKL function and substrate(s), as well as its contribution to the retinal etiopathology, remain as yet unknown. In this work we aimed to characterize the mouse retinal sphingolipidome by UPLC-TOF to first, thoroughly investigate the SPL composition of the murine retina, compare it to our Cerkl -/- model, and finally assess new possible CERKL substrates by phosphorus quantification and protein-lipid overlay. Our results showed a consistent and notable decrease of the retinal SPL content (mainly ranging from 30% to 60%) in the Cerkl -/- compared to WT retinas, which was particularly evident in the glucosyl/galactosyl ceramide species (Glc/GalCer) whereas the phospholipids and neutral lipids remained unaltered. Moreover, evidence in favor of CERKL binding to GlcCer, GalCer and sphingomyelin has been gathered. Altogether, these results highlight the involvement of CERKL in the SPL metabolism, question its role as a kinase, and open new scenarios concerning its function.


Increased spinal adenosine impairs phrenic long-term facilitation in aging rats.
Moderate acute intermittent hypoxia (mAIH) elicits a form of spinal, respiratory motor plasticity known as phrenic long-term facilitation (pLTF). In middle-aged male and geriatric female rats, mAIH-induced pLTF is attenuated through unknown mechanisms. In young adults, mAIH activates competing intracellular signaling cascades, initiated by serotonin 2 and adenosine 2A (A2A) receptors, respectively. Spinal A2A receptor inhibition enhances mAIH-induced pLTF, meaning, serotonin dominates, and adenosine constrains mAIH-induced plasticity in the daily rest phase. Thus, we hypothesized elevated basal adenosine levels in the ventral cervical spinal cord of aged rats shifts this balance, undermining mAIH-induced pLTF. A selective A2A receptor antagonist (MSX-3) or vehicle was delivered intrathecally at C4 in anesthetized young (3-6 mo) and aged (20-22 mo) Sprague-Dawley rats before mAIH (3,5-min episodes; arterial Po2 = 45-55 mmHg). In young males, spinal A2A receptor inhibition enhanced pLTF (119 ± 5%) vs. vehicle (55 ± 9%), consistent with prior reports. In old males, pLTF was reduced to 25 ± 11%, but A2A receptor inhibition increased pLTF to levels greater than in young males (186 ± 19%). Basal adenosine levels in ventral C3-C5 homogenates are elevated two- to threefold in old vs. young males. These findings advance our understanding of age as a biological variable in phrenic motor plasticity and will help guide translation of mAIH as a therapeutic modality to restore respiratory and nonrespiratory movements in older populations afflicted with clinical disorders that compromise movement.NEW & NOTEWORTHY Advanced age undermines respiratory motor plasticity, specifically phrenic long-term facilitation (pLTF) following moderate acute intermittent hypoxia (mAIH). We report that spinal adenosine increases in aged male rats, undermining mAIH-induced pLTF via adenosine 2A (A2A) receptor activation, an effect reversed by selective spinal adenosine 2A receptor inhibition. These findings advance our understanding of mechanisms that impair neuroplasticity, and the ability to compensate for the onset of lung or neural injury with age, and may guide efforts to harness mAIH as a treatment for clinical disorders that compromise breathing and other movements.


Comparative expression profiling in grape (Vitis vinifera) berries derived from frequency analysis of ESTs and MPSS signatures.
Vitis vinifera (V. vinifera) is the primary grape species cultivated for wine production, with an industry valued annually in the billions of dollars worldwide. In order to sustain and increase grape production, it is necessary to understand the genetic makeup of grape species. Here we performed mRNA profiling using Massively Parallel Signature Sequencing (MPSS) and combined it with available Expressed Sequence Tag (EST) data. These tag-based technologies, which do not require a priori knowledge of genomic sequence, are well-suited for transcriptional profiling. The sequence depth of MPSS allowed us to capture and quantify almost all the transcripts at a specific stage in the development of the grape berry. The number and relative abundance of transcripts from stage II grape berries was defined using Massively Parallel Signature Sequencing (MPSS). A total of 2,635,293 17-base and 2,259,286 20-base signatures were obtained, representing at least 30,737 and 26,878 distinct sequences. The average normalized abundance per signature was approximately 49 TPM (Transcripts Per Million). Comparisons of the MPSS signatures with available Vitis species' ESTs and a unigene set demonstrated that 6,430 distinct contigs and 2,190 singletons have a perfect match to at least one MPSS signature. Among the matched sequences, ESTs were identified from tissues other than berries or from berries at different developmental stages. Additional MPSS signatures not matching to known grape ESTs can extend our knowledge of the V. vinifera transcriptome, particularly when these data are used to assist in annotation of whole genome sequences from Vitis vinifera. The MPSS data presented here not only achieved a higher level of saturation than previous EST based analyses, but in doing so, expand the known set of transcripts of grape berries during the unique stage in development that immediately precedes the onset of ripening. The MPSS dataset also revealed evidence of antisense expression not previously reported in grapes but comparable to that reported in other plant species. Finally, we developed a novel web-based, public resource for utilization of the grape MPSS data [1].


Survival predictors in paraquat intoxification and role of immunosuppression.
Paraquat poisoning resulted in multiorgan failure and is associated with high mortality. We audited 83 historical cases of paraquat poisoning in past 2 years treated with conventional decontamination and supportive treatment, followed by enrolling 85 patients over a 2 year period into additional immunosuppression with intravenous (i.v.) methylprednisolone and i.v. cyclophosphamide. Our results showed that age, poor renal function and leucocytosis are the main predictors of fatal outcome. Immunosuppression regime rendered higher survival (6 out of 17 patients (35.3%)) versus historical control (1 out of 18 patients (5.6%)) (p = 0.041) in the cohort with admission eGFR < 50 ml/min/1.73 m2 and WBC count > 11,000/μL. In contrast, there was no difference in survival with immunosuppression regime (38 out of 64 patients (59.4%)) compared to historical control (30 out of 52 patients (57.7%)) (p = 0.885) in those with eGFR > 50 ml/min/1.73 m2 or WBC < 11,000/μL at presentation. Multivariable logistic regression showed survival probability = exp(logit)/(1 + exp(logit)), in which logit = 13.962 - (0.233 × ln(age (year))) - (1.344 × ln(creatinine (μmol/L))) - (1.602 × ln(rise in creatinine (μmol/day))) - (0.614 × ln(WBC (,000/μL))) + (2.021 × immunosuppression) and immunosuppression = 1 if given and 0 if not. Immunosuppression therapy yielded odds ratio of 0.132 (95% confidential interval: 0.029-0.603, p = 0.009). In conclusion, immunosuppression therapy with intravenous methylprednisolone and cyclophosphamide may counteract immune mediated inflammation after paraquat poisoning and improve survival of patients with admission eGFR < 50 ml/min/1.73 m2 and WBC count > 11,000/μL.