Transcranial direct current stimulation (tDCS) of frontal cortex decreases performance on the WAIS-IV intelligence test.
Transcranial direct current stimulation (tDCS) modulates excitability of motor cortex. However, there is conflicting evidence about the efficacy of this non-invasive brain stimulation modality to modulate performance on cognitive tasks. Previous work has tested the effect of tDCS on specific facets of cognition and executive processing. However, no randomized, double-blind, sham-controlled study has looked at the effects of tDCS on a comprehensive battery of cognitive processes. The objective of this study was to test if tDCS had an effect on performance on a comprehensive assay of cognitive processes, a standardized intelligence quotient (IQ) test. The study consisted of two substudies and followed a double-blind, between-subjects, sham-controlled design. In total, 41 healthy adult participants were included in the final analysis. These participants completed the Wechsler Adult Intelligence Scale, Fourth Edition (WAIS-IV) as a baseline measure. At least one week later, participants in substudy 1 received either bilateral tDCS (anodes over both F4 and F3, cathode over Cz, 2 mA at each anode for 20 min) or active sham tDCS (2 mA for 40 s), and participants in substudy 2 received either right or left tDCS (anode over either F4 or F3, cathode over Cz, 2 mA for 20 min). In both studies, the WAIS-IV was immediately administered following stimulation to assess for performance differences induced by bilateral and unilateral tDCS. Compared to sham stimulation, right, left, and bilateral tDCS reduced improvement between sessions on Full Scale IQ and the Perceptual Reasoning Index. This demonstration that frontal tDCS selectively degraded improvement on specific metrics of the WAIS-IV raises important questions about the often proposed role of tDCS in cognitive enhancement.


An Isolated Transosseous Flexible Suture Frame in the Treatment of Patellar Tendon Rupture Provides Adequate Mechanical Resistance.
Acute patellar tendon ruptures are frequently observed in patients with metabolic comorbidities, and the benchmark treatment is surgical repair. It is desirable not to harm an already fragile biologic environment with sutures and hardware. We aimed to compare the mechanical requirements of an isolated, flexible, high-strength nonabsorbable transosseous suture frame with that of the Krackow suture technique. A total of 12 cadaveric pieces were randomized into two groups: the isolated flexible frame group (n = 6) and the standard Krackow fixation group (n = 6). A traumatic rupture of the patellar tendon was performed, and a transosseous displacement sensor was installed on a validated biomechanical system. Gap formation was measured during 50 cycles of flexion and extension with traction on the quadriceps (250 N). Subsequently, specimens underwent progressive loading in a fixed flexion position until failure occurred. The data were analyzed using nonparametric statistical tools with a significance level of 5%. The isolated frame group had a smaller gap formation (1.7 mm) than the Krackow group (3.4 mm; P = 0.01). No significant difference existed in the median failure end points of the two groups (676 and 530 N, respectively; P = 0.11). Patellar tendon repair using an isolated, transosseous, flexible, suture frame outperformed using the traditional Krakow repair technique in gap formation. Further studies are needed to determine if this will result in better functional outcomes or fewer clinical failures. Level IV, experimental case series.


Cutting edge: suppression of GM-CSF expression in murine and human T cells by IL-27.
GM-CSF is a potent proinflammatory cytokine that plays a pathogenic role in the CNS inflammatory disease experimental autoimmune encephalomyelitis. As IL-27 alleviates experimental autoimmune encephalomyelitis, we hypothesized that IL-27 suppresses GM-CSF expression by T cells. We found that IL-27 suppressed GM-CSF expression in CD4+ and CD8+ T cells in splenocyte and purified T cell cultures. IL-27 suppressed GM-CSF in Th1, but not Th17, cells. IL-27 also suppressed GM-CSF expression by human T cells in nonpolarized and Th1- but not Th17-polarized PBMC cultures. In vivo, IL-27p28 deficiency resulted in increased GM-CSF expression by CNS-infiltrating T cells during Toxoplasma gondii infection. Although in vitro suppression of GM-CSF by IL-27 was independent of IL-2 suppression, IL-10 upregulation, or SOCS3 signaling, we observed that IL-27-driven suppression of GM-CSF was STAT1 dependent. Our findings demonstrate that IL-27 is a robust negative regulator of GM-CSF expression in T cells, which likely inhibits T cell pathogenicity in CNS inflammation.


Reduced sensitivity of SARS-CoV-2 variant Delta to antibody neutralization.
The SARS-CoV-2 B.1.617 lineage was identified in October 2020 in India1-5. Since then, it has become dominant in some regions of India and in the UK, and has spread to many other countries6. The lineage includes three main subtypes (B1.617.1, B.1.617.2 and B.1.617.3), which contain diverse mutations in the N-terminal domain (NTD) and the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein that may increase the immune evasion potential of these variants. B.1.617.2-also termed the Delta variant-is believed to spread faster than other variants. Here we isolated an infectious strain of the Delta variant from an individual with COVID-19 who had returned to France from India. We examined the sensitivity of this strain to monoclonal antibodies and to antibodies present in sera from individuals who had recovered from COVID-19 (hereafter referred to as convalescent individuals) or who had received a COVID-19 vaccine, and then compared this strain with other strains of SARS-CoV-2. The Delta variant was resistant to neutralization by some anti-NTD and anti-RBD monoclonal antibodies, including bamlanivimab, and these antibodies showed impaired binding to the spike protein. Sera collected from convalescent individuals up to 12 months after the onset of symptoms were fourfold less potent against the Delta variant relative to the Alpha variant (B.1.1.7). Sera from individuals who had received one dose of the Pfizer or the AstraZeneca vaccine had a barely discernible inhibitory effect on the Delta variant. Administration of two doses of the vaccine generated a neutralizing response in 95% of individuals, with titres three- to fivefold lower against the Delta variant than against the Alpha variant. Thus, the spread of the Delta variant is associated with an escape from antibodies that target non-RBD and RBD epitopes of the spike protein.


Variation within the type 2 diabetes susceptibility gene calpain-10 and polycystic ovary syndrome.
Variation within the calpain-10 gene (CAPN10) has been proposed to account for linkage to type 2 diabetes on chromosome 2q in Mexican-Americans, and associations with diabetes have been reported in several other populations. Given the epidemiological, physiological, and genetic overlap between type 2 diabetes and polycystic ovary syndrome (PCOS), CAPN10 represents a strong candidate gene for a role in PCOS susceptibility. Using both family based and case-control association resources (146 parent-offspring trios; 185 additional PCOS cases; 525 control subjects, all of European ancestry), we sought association between CAPN10 variation and PCOS, focusing on four single nucleotide polymorphism (SNP) variants (SNP-44, SNP-43; SNP-19; SNP-63). On single-locus transmission disequilibrium analysis in the 146 trios, there was nominal evidence (P = 0.03) of excess transmission of the more common allele at SNP-63. This association was not, however, replicated in the case-control analysis. No other significant associations were observed at the single-locus or haplotype level in either the transmission-disequilibrium or case-control analyses. The relative risk for the high-risk diabetes susceptibility 112/121 genotype (SNPs 43-19-63) was 0.84 (95% confidence intervals, 0.40-1.71). No associations were seen with intermediate traits of relevance to diabetes and PCOS pathogenesis. We have found no evidence from these analyses that CAPN10 gene variation influences susceptibility to PCOS.


Complementary and alternative therapies among very long-term breast cancer survivors.
Breast cancer patients may have different complementary and alternative medicine (CAM) usage rates and may turn to CAM for different reasons than healthy adults. CAM has mostly been studied in recently diagnosed women; no studies have included survivors 10 years post-diagnosis. We examined very long-term breast cancer survivors to determine whether CAM users had dissimilar patterns of association with survivorship factors. Interviews of 374 breast cancer case patients from a population-based case-control breast cancer study of young women from Los Angeles County, California, during the 1980s occurred at follow-up; 371 patients with complete information were included. CAM represented 28 herbal remedies. Quality-of-life originated from the Medical Outcomes Study Short Form 36 questionnaire (SF-36). Higher rates of CAM (59%) usage occurred compared to nationwide estimates. CAM users resembled non-users on follow-up age, exercise, original disease, treatment, smoking, body-mass index, alcohol, and fear of recurrence. CAM users had a higher prevalence of medical co-morbidities (P = 0.0005), and scored significantly lower on the SF-36 emotional well-being subscale than non-CAM users (P = 0.01). CAM users and non-users did not differ on the SF-36 physical sub-scale. Very long-term breast cancer survivors who use CAM may have poorer emotional functioning and more medical problems than non-users.


"Employment and arthritis: making it work" a randomized controlled trial evaluating an online program to help people with inflammatory arthritis maintain employment (study protocol).
Arthritis and musculoskeletal conditions are the leading cause of long-term work disability (WD), an outcome with a major impact on quality of life and a high cost to society. The importance of decreased at-work productivity has also recently been recognized. Despite the importance of these problems, few interventions have been developed to reduce the impact of arthritis on employment. We have developed a novel intervention called "Making It Work", a program to help people with inflammatory arthritis (IA) deal with employment issues, prevent WD and improve at-work productivity. After favorable results in a proof-of-concept study, we converted the program to a web-based format for broader dissemination and improved accessibility. The objectives of this study are: 1) to evaluate in a randomized controlled trial (RCT) the effectiveness of the program at preventing work cessation and improving at-work productivity; 2) to perform a cost-utility analysis of the intervention. 526 participants with IA will be recruited from British Columbia, Alberta, and Ontario in Canada. The intervention consists of a) 5 online group sessions; b) 5 web-based e-learning modules; c) consultations with an occupational therapist for an ergonomic work assessment and a vocational rehabilitation counselor. Questionnaires will be administered online at baseline and every 6 months to collect information about demographics, disease measures, costs, work-related risk factors for WD, quality of life, and work outcomes. Primary outcomes include at-work productivity and time to work cessation of > 6 months for any reason. Secondary outcomes include temporary work cessation, number of days missed from work per year, reduction in hours worked per week, quality adjusted life year for the cost utility analysis, and changes from baseline in employment risk factors. Analysis of Variance will evaluate the intervention's effect on at-work productivity, and multivariable Cox regression models will estimate the risk of work cessation associated with the intervention after controlling for risk factors for WD and other important predictors imbalanced at baseline. This program fills an important gap in arthritis health services and addresses an important and costly problem. Knowledge gained from the RCT will be useful to health care professionals, policy planners and arthritis stakeholders. ClinicalTrials.gov NCT01852851; registered April 13, 2012; first participant randomized on July 6, 2013.


An integrated approach to diagnosing and managing sleep disorders in menopausal women.
Sleep disorders increase in prevalence during the menopausal transition and they constitute a complex phenomenon. Insomnia, the main sleep disorder, can be a primary disorder or it can be secondary to hot flushes (HF), mood disorders, psychosocial factors, medical conditions, and other sleep disturbances, such as obstructive sleep apnoea (OSA) or restless legs syndrome (RLS). Menopausal women complaining of persistent sleep disorders should be referred to a sleep specialist for comprehensive sleep management because unrecognized and untreated sleep disorders can have dramatic health-related consequences. Women suffering from insomnia related to vasomotor symptoms (VMS) can be treated with hormone replacement therapy (HRT). Primary insomnia will be preferentially improved with cognitive behavioural therapy (CBT-I) or with non-benzodiazepine hypnotics or melatonin. CBT-I is a highly efficacious treatment for postmenopausal women with insomnia. Using antidepressants to treat sleep disruption in the absence of depression is not recommended; instead, the United States Food and Drug Administration (FDA) approved paroxetine as the first non-hormonal treatment for HF. Sleep disorders in menopausal women should not be underestimated. It is necessary to diagnose the specific causal disorder and then to provide treatment to improve sleep quality and quality of life.


Motivations for entry into sex work and HIV risk among mobile female sex workers in India.
This paper assesses the reasons for entry into sex work and its association with HIV risk behaviours among mobile female sex workers (FSWs) in India. Data were collected from a cross-sectional survey conducted in 22 districts across four high HIV prevalence states in India during 2007-2008. Analyses were limited to 5498 eligible mobile FSWs. The reasons given by FSWs for entering sex work and associations with socio-demographic characteristics were assessed. Reported reasons for entering sex work include poor or deprived economic conditions; negative social circumstances in life; own choice; force by an external person; and family tradition. The results from multivariate analyses indicate that those FSWs who entered sex work due to poor economic conditions or negative social circumstances in life or force demonstrated elevated levels of current inconsistent condom use as well as in the past in comparison with those FSWs who reported entering sex work by choice or family tradition. This finding indicates the need for a careful assessment of the pre-entry contexts among HIV prevention interventions since these factors may continue to hinder the effectiveness of efforts to reduce the spread of HIV/AIDS in India and elsewhere.


Inhibition of proteasome deubiquitinase activity: a strategy to overcome resistance to conventional proteasome inhibitors?
Although more traditionally associated with degradation and maintenance of protein homeostasis, the ubiquitin-proteasome system (UPS) has emerged as a critical component in the regulation of cancer cell growth and survival. The development of inhibitors that block the proteolytic activities of the proteasome have highlighted its suitability as a bona fide anti-cancer drug target. However, key determinants including the development of drug resistance and dose-limiting toxicity call for the identification of alternative components of the UPS for novel drug targeting. Recently the deubiquitinases (DUBs), a diverse family of enzymes that catalyze ubiquitin removal, have attracted significant interest as targets for the development of next generation UPS inhibitors. In particular, pharmacological inhibition of the proteasomal cysteine DUBs (i.e., USP14 and UCHL5) has been shown to be particularly cytotoxic to cancer cells and inhibit tumour growth in several in vivo models. In the current review we focus on the modes of action of proteasome DUB inhibitors and discus the potential of DUB inhibitors to circumvent acquired drug resistance and provide a therapeutic option for the treatment of cancer.


Enabling continuous learning and quality improvement in health care.
Purpose The purpose of this paper is to examine how the design and implementation of learning models for performance management can foster continuous learning and quality improvement within a publicly funded, multi-site community hospital organization. Design/methodology/approach Niagara Health's patient flow performance management system, a learning model, was studied over a 20-month period. A descriptive case study design guided the analysis of qualitative observational data and its synthesis with organizational learning theory literature. Emerging from this analysis were four propositions to inform the implementation of learning models and future research. Findings This performance management system was observed to enable: ongoing performance-related knowledge exchange by creating opportunities for routine social interaction; collective recognition and understanding of practice and performance patterns; relationship building, learning for improvement, and "higher order" learning through dialogue facilitated using humble inquiry; and, alignment of quality improvement efforts to organizational strategic objectives through a multi-level feedback/feed-forward communication structure. Research limitations/implications The single organization and descriptive study design may limit the generalizability of the findings and introduce confirmation bias. Future research should more comprehensively evaluate the impact of learning models on organizational learning processes and performance outcomes. Practical implications This study offers novel insight which may inform the design and implementation of learning models for performance management within and beyond the study site. Originality/value Few studies have examined the mechanics of performance management systems in relation to organizational learning theory and research. Broader adoption of learning models may be key to the development of continuously learning and improving health systems.


Upregulation of indoleamine 2,3-dioxygenase in hepatitis C virus infection.
Indoleamine 2,3-dioxygenase (IDO) is induced by proinflammatory cytokines and by CTLA-4-expressing T cells and constitutes an important mediator of peripheral immune tolerance. In chronic hepatitis C, we found upregulation of IDO expression in the liver and an increased serum kynurenine/tryptophan ratio (a reflection of IDO activity). Huh7 cells supporting hepatitis C virus (HCV) replication expressed higher levels of IDO mRNA than noninfected cells when stimulated with gamma interferon or when cocultured with activated T cells. In infected chimpanzees, hepatic IDO expression decreased in animals that cured the infection, while it remained high in those that progressed to chronicity. For both patients and chimpanzees, hepatic expression of IDO and CTLA-4 correlated directly. Induction of IDO may dampen T-cell reactivity to viral antigens in chronic HCV infection.


Uptake of all-trans retinoic acid-containing aerosol by inhalation to lungs in a guinea pig model system--a pilot study.
Systemic therapies with retinoic acid (RA) can result in toxic side effects without yielding biologically effective levels in target tissues such as lung. The authors adapted a PARI LC Star nebulizer to create a tubular system for short-term inhalation treatment of guinea pigs using a water-miscible formulation of all-trans RA (ATRA) or vehicle. Based on the initial average weight, animals received an estimated average ATRA doses of either 0.32 mg·kg(-1) (low dose, 1.4 mM), or 0.62 mg·kg(-1) (medium dose, 2.8 mM), or 1.26 mg·kg(-1) (high dose, 5.6 mM) 20 minutes per day for 6 consecutive days. This system led to a rise of ATRA levels in lung, but not liver or plasma. Cellular lung levels of retinol, retinyl palmitate, and retinyl stearate also appeared to be unaffected (245.6 ± 10.7, 47.4 ± 3.4, and 132.8 ± 7.7 ng·g(-1) wet weight, respectively). The application of this aerosolized ATRA also induced a dose-dependent protein expression of the cellular retinol-binding protein 1 (CRBP-1) in lung, without apparent harmful side effects.


A bone sialoprotein-binding protein from Staphylococcus aureus: a member of the staphylococcal Sdr family.
Staphylococcus aureus bacteria, isolated from bone and joint infections, specifically interact with bone sialoprotein (BSP), a glycoprotein of bone and dentine extracellular matrix, via a cell-surface protein of M(r) 97000 [Yacoub, Lindahl, Rubin, Wendel, Heinegârd and Rydén, (1994) Eur. J. Biochem. 222, 919-925]. Amino acid sequences of seven trypsin fragments from the 97000-M(r) BSP-binding protein were determined. A gene encoding a protein encompassing all seven peptide sequences was identified from chromosomal DNA isolated from S. aureus strain O24. This gene encodes a protein with 1171 amino acids, called BSP-binding protein (Bbp), which displays similarity to recently described proteins of the Sdr family from S. aureus. SdrC, SdrD and SdrE encode putative cell-surface proteins with no described ligand specificity. Bbp also shows similarity to a fibrinogen-binding protein from S. epidermidis called Fbe. A serine-aspartic acid repeat sequence was found close to the cell-wall-anchoring Leu-Pro-Xaa-Thr-Gly sequence in the C-terminal end of the protein. Escherichia coli cells were transformed with an expression vector containing a major part of the bbp gene fused to the gene for glutathione S-transferase. The affinity-purified fusion protein bound radiolabelled native BSP, and inhibited the binding of radiolabelled BSP to staphylococcal cells. Serum from patients suffering from bone and joint infection contained antibodies that reacted with the fusion protein of the BSP-binding protein, indicating that the protein is expressed during an infection and is immunogenic. The S. aureus Bbp protein may be important in the localization of bacteria to bone tissue, and thus might be of relevance in the pathogenicity of osteomyelitis.


Not all dyslexics are created equal.
Dyslexia is a common disorder that has traditionally been treated as a homogeneous condition. However, recent evidence indicates that it is a heterogenous condition with several subtypes. For example, studies of the visual system indicate that not all dyslexics have a normal visual pathway. Approximately 75% have a processing deficit in the magnocellular pathway. Our previous study indicated that dysphoneidetic but not dyseidetic dyslexics exhibit a magnocellular pathway defect. The purpose of this study was to expand our previous work by also examining dysphonetic dyslexics. Additionally, the stimulus was altered to enhance detection of a magnocellular pathway defect in any dyslexic subtype. Temporal contrast sensitivity functions were determined with a flickering stimulus (5, 10, 15, 20, and 25 Hz) by using a temporal, two-alternative, forced-choice technique. The results indicate that the dyseidetic dyslexics do not have a magnocellular pathway defect, whereas the dysphoneidetics do. Furthermore, examination of the individual dysphonetics indicated that the more severely affected subjects also exhibited a magnocellular pathway defect. These results suggest that treatment strategies for dyslexics may need to be modified to take into account their specific subtype.


Executive and memory correlates of age-related differences in wayfinding performances using a virtual reality application.
The aim of this study was to evaluate in large-scale spaces wayfinding and spatial learning difficulties for older adults in relation to the executive and memory decline associated with aging. We compared virtual reality (VR)-based wayfinding and spatial memory performances between young and older adults. Wayfinding and spatial memory performances were correlated with classical measures of executive and visuo-spatial memory functions, but also with self-reported estimates of wayfinding difficulties. We obtained a significant effect of age on wayfinding performances but not on spatial memory performances. The overall correlations showed significant correlations between the wayfinding performances and the classical measures of both executive and visuo-spatial memory, but only when the age factor was not partialled out. Also, older adults underestimated their wayfinding difficulties. A significant relationship between the wayfinding performances and self-reported wayfinding difficulty estimates is found, but only when the age effect was partialled out. These results show that, even when older adults have an equivalent spatial knowledge to young adults, they had greater difficulties with the wayfinding task, supporting an executive decline view in age-related wayfinding difficulties. However, the correlation results are in favor of both the memory and executive decline views as mediators of age-related differences in wayfinding performances. This is discussed in terms of the relationships between memory and executive functioning in wayfinding task orchestration. Our results also favor the use of objective assessments of everyday navigation difficulties in virtual applications, instead of self-reported questionnaires, since older adults showed difficulties in estimating their everyday wayfinding problems.


Dominance and submission: the ventrolateral prefrontal cortex and responses to status cues.
Status hierarchies constitute a fundamental organizing principle of human society. However, little is known about the neural systems that process nonverbal cues that indicate status. Preliminary neuropsychological work has suggested a role for the ventrolateral and ventromedial prefrontal cortex (VLPFC/VMPFC) and the superior temporal cortex (STC). We used functional magnetic resonance imaging to delineate the nature of these roles. Analyses revealed signal changes in the right VLPFC in connection with two primary functions attributed to status cues. Status cues moderate behavior and the right VLPFC showed increased signal for high-status relative to neutral and low-status cues. The VLPFC also showed increased signal for high-status cues displayed by individuals of the opposite gender to the perceiver; this may be relevant to the role status cues play in moderating mate choice behavior. Connectivity results indicated significant positive connectivity between the VLPFC and both the VMPFC and the STC. We suggest that the VLPFC retrieves information from these regions when processing hierarchy cues to facilitate socially adaptive behavior.


Therapeutic effects of Low intensity extracorporeal low energy shock wave therapy (LiESWT) on stress urinary incontinence.
This study aimed to evaluate the therapeutic effects of Low intensity extracorporeal low energy shock wave therapy (LiESWT) on stress urinary incontinence (SUI). The investigation was a single-arm, open-label, multicentre study conducted in Taiwan. 50 female patients with SUI received LiESWT-treated with 0.25 mJ/mm2 intensity, 3000 pulses, and 3 pulses/second, once weekly for 4-weeks (W4) and 8-weeks (W8). The pad test, uroflowmetry, life quality questionnaires, and 3-day urinary diary measurement were performed before and after LiESWT intervention. The results revealed that 8-week of LiESWT treatment meaningfully improved urine leakage (pad test), maximum flow rate, post-voided residual urine, average urine volume, functional bladder capacity, urinary frequency, urgency symptom, and nocturia, which also persisted to show significant improvements at 1-month follow up (F1). Moreover, bothersome questionnaires scores were significantly improved at W4, W8, and F1 as compared to the baseline (W0). These results indicated that 8 weeks of LiESWT attenuated SUI symptoms on physical activity, reduced bladder leaks and overactive bladder (OAB), implying that LiESWT brought significant improvement in the quality of life. (ClinicalTrials.gov number, NCT04059133).


Fine-Mapping of Common Genetic Variants Associated with Colorectal Tumor Risk Identified Potential Functional Variants.
Genome-wide association studies (GWAS) have identified many common single nucleotide polymorphisms (SNPs) associated with colorectal cancer risk. These SNPs may tag correlated variants with biological importance. Fine-mapping around GWAS loci can facilitate detection of functional candidates and additional independent risk variants. We analyzed 11,900 cases and 14,311 controls in the Genetics and Epidemiology of Colorectal Cancer Consortium and the Colon Cancer Family Registry. To fine-map genomic regions containing all known common risk variants, we imputed high-density genetic data from the 1000 Genomes Project. We tested single-variant associations with colorectal tumor risk for all variants spanning genomic regions 250-kb upstream or downstream of 31 GWAS-identified SNPs (index SNPs). We queried the University of California, Santa Cruz Genome Browser to examine evidence for biological function. Index SNPs did not show the strongest association signals with colorectal tumor risk in their respective genomic regions. Bioinformatics analysis of SNPs showing smaller P-values in each region revealed 21 functional candidates in 12 loci (5q31.1, 8q24, 11q13.4, 11q23, 12p13.32, 12q24.21, 14q22.2, 15q13, 18q21, 19q13.1, 20p12.3, and 20q13.33). We did not observe evidence of additional independent association signals in GWAS-identified regions. Our results support the utility of integrating data from comprehensive fine-mapping with expanding publicly available genomic databases to help clarify GWAS associations and identify functional candidates that warrant more onerous laboratory follow-up. Such efforts may aid the eventual discovery of disease-causing variant(s).


Novel HDAC inhibitor MAKV-8 and imatinib synergistically kill chronic myeloid leukemia cells via inhibition of BCR-ABL/MYC-signaling: effect on imatinib resistance and stem cells.
Chronic myeloid leukemia (CML) pathogenesis is mainly driven by the oncogenic breakpoint cluster region-Abelson murine leukemia viral oncogene homolog 1 (BCR-ABL) fusion protein. Since BCR-ABL displays abnormal constitutive tyrosine kinase activity, therapies using tyrosine kinase inhibitors (TKis) such as imatinib represent a major breakthrough for the outcome of CML patients. Nevertheless, the development of TKi resistance and the persistence of leukemia stem cells (LSCs) remain barriers to cure the disease, justifying the development of novel therapeutic approaches. Since the activity of histone deacetylase (HDAC) is deregulated in numerous cancers including CML, pan-HDAC inhibitors may represent promising therapeutic regimens for the treatment of CML cells in combination with TKi. We assessed the anti-leukemic activity of a novel hydroxamate-based pan-HDAC inhibitor MAKV-8, which complied with the Lipinski's "rule of five," in various CML cells alone or in combination with imatinib. We validated the in vitro HDAC-inhibitory potential of MAKV-8 and demonstrated efficient binding to the ligand-binding pocket of HDAC isoenzymes. In cellulo, MAKV-8 significantly induced target protein acetylation, displayed cytostatic and cytotoxic properties, and triggered concomitant ER stress/protective autophagy leading to canonical caspase-dependent apoptosis. Considering the specific upregulation of selected HDACs in LSCs from CML patients, we investigated the differential toxicity of a co-treatment with MAKV-8 and imatinib in CML versus healthy cells. We also showed that beclin-1 knockdown prevented MAKV-8-imatinib combination-induced apoptosis. Moreover, MAKV-8 and imatinib co-treatment synergistically reduced BCR-ABL-related signaling pathways involved in CML cell growth and survival. Since our results showed that LSCs from CML patients overexpressed c-MYC, importantly MAKV-8-imatinib co-treatment reduced c-MYC levels and the LSC population. In vivo, tumor growth of xenografted K-562 cells in zebrafish was completely abrogated upon combined treatment with MAKV-8 and imatinib. Collectively, the present findings show that combinations HDAC inhibitor-imatinib are likely to overcome drug resistance in CML pathology.


Compression of digital chest radiographs with a mixture of principal components neural network: evaluation of performance.
The performance of a new, neural network-based image compression method was evaluated on digital radiographs for use in an educational environment. The network uses a mixture of principal components (MPC) representation to effect optimally adaptive transform coding of an image and has significant computational advantages over other techniques. Nine representative digital chest radiographs were compressed 10:1, 20:1, 30:1, and 40:1 with the MPC method. The five versions of each image, including the original, were shown simultaneously, in random order, to each of seven radiologists, who rated each one on a five-point scale for image quality and visibility of pathologic conditions. One radiologist also ranked four versions of each of the nine images in terms of the severity of distortion: The four versions represented 30:1 and 40:1 compression with the MPC method and with the classic Karhunen-Loève transform (KLT). Only for the images compressed 40:1 with the MPC method were there any unacceptable ratings. Nevertheless, the images compressed 40:1 received a top score in 26%-33% of the evaluations. Images compressed with the MPC method were rated better than or as good as images compressed with the KLT technique 17 of 18 times. Four of nine times, images compressed 40:1 with the MPC method were rated as good as or better than images compressed 30:1 with the KLT technique.


Microwave photonic quadrature filter based on an all-optical programmable Hilbert transformer.
A microwave photonic quadrature filter, new to our knowledge, based on an all-optical Hilbert transformer is presented. It is based on mapping of a Hilbert transform transfer function between the optical and electrical domains, using a programmable Fourier-domain optical processor and high-speed photodiodes. The technique enables the realization of an extremely wide operating bandwidth, tunable programmable bandwidth, and a highly precise amplitude and phase response. Experimental results demonstrate a microwave quadrature filter from 10 to 20 GHz, which achieves an amplitude imbalance of less than ±0.23 dB and a phase imbalance of less than ±0.5°.


Dyspareunia and urinary sensory symptoms in India: population-based study.
Population level estimates of sexual dysfunction in less developed settings where sexuality is not openly discussed or is culturally constrained are lacking. To determine the prevalence of dyspareunia and identify associated symptoms and sociocultural factors. Data from a population-based national level family health sample survey conducted in Indian states from 1998 to 1999 which collected sexual and reproductive health information from 84,644 currently married women. The main outcome measure was dyspareunia. The overall prevalence of dyspareunia was 12.6%, with a higher prevalence in the central region, among newly married and younger women, among Muslims, rural residents, and among nonusers or traditional contraceptive method users. Dyspareunia was significantly more common among respondents who had urinary sensory symptoms when compared with their counterparts (adjusted odds ratio: 6.57, 95% confidence interval: 6.28, 6.87). Dyspareunia prevalence and the associated symptoms reported in this analysis could be underestimates because of possible underreporting. There is a substantial hidden burden of sexual health problems especially affecting younger women.


Diagnosis and management of fragile X syndrome.
To complement the 2005 Annual Clinical Focus on medical genomics, AFP will be publishing a series of short reviews on genetic syndromes. This series was designed to increase awareness of these diseases so that family physicians can recognize and diagnose children with these disorders and understand the kind of care they might require in the future. The first review in this series discusses fragile X syndrome.


Possible role of NF-kappaB and p53 in the glutamate-induced pro-apoptotic neuronal pathway.
Apoptosis is now recognized as an important component in many progressive and acute neurodegenerative diseases. Extracellular signals and intracellular mechanisms triggering and regulating apoptosis in neuronal cells are still a matter of investigation. Here we review data from our and other laboratories with the aim to elucidate the nature of some proteins which are known to be involved in cell cycle regulation as well as in promoting degeneration and apoptosis of neurons. The following molecules will be taken into consideration: NF-kappaB, p53, p21 and MSH2. These proteins are activated by neurotoxic experimental conditions which involve the stimulation of selective receptors for the excitatory aminoacid glutamate. Thus, we hypothesize their contribution to an intracellular pathway responsible for the glutamate-induced neuronal death. Identification of such mechanisms could be relevant for understanding the apoptosis associated with various neurodegenerative diseases as well as for developing novel strategies of pharmacological intervention.


A randomised controlled cross-over trial of aerobic training versus Qigong in advanced Parkinson's disease.
To investigate the effects of an aerobic training in subjects with Parkinson's disease (PD) as compared to a medical Chinese exercise (Qigong). randomized controlled trial with a cross over design. PD out-patients referred to a Neurorehabilitation facility for the management of motor disability. 26 PD patients in Hoehn and Yahr stage II to III under stable medication were randomly allocated to either Group AT1+QG2 (receiving 20 aerobic training sessions followed by 20 ''Qigong'' group sessions with 2 month interval between the interventions), or Group QG1+AT2 (performing the same treatments with an inverted sequence). clinical effects of treatment were sought through the Unified Parkinson's Disease Rating Scale (UPDRS), Brown's Disability Scale (B'DS), six-Minute Walking Test (6MWT), Borg scale for breathlessness, Beck Depression Inventory (BDI) and Parkinson's Disease Questionnaire-39 items (PDQ-39). A spirometry test and maximum cardiopulmonary exercise test (CPET) were also performed to determine the pulmonary function, the metabolic and cardio-respiratory requests at rest and under exercise. All measures were taken immediately before and at the completion of each treatment phase. The statistical analysis focusing on the evolution of motor disability and quality of life revealed a significant interaction effect between group and time for the 6MWT (time x group effect: F: 5.4 P=0.002) and the Borg scale (time x group effect: F: 4.2 P=0.009). Post hoc analysis showed a significant increase in 6MWT and a larger decrease in Borg score after aerobic training within each subgroup, whereas no significant changes were observed during Qigong. No significant changes over time were detected through the analysis of UPDRS, B'DS, BDI and PDQ-39 scores. The analysis of cardiorespiratory parameters showed significant interaction effects between group and time for the Double Productpeak (time x group effect: F: 7.7 P=0.0003), the VO(2peak) (time x group effect: F: 4.8 P=0.007), and the VO(2)/kg ratio (time x group effect: F: 4.3 P=0.009), owing to their decrease after aerobic training to an extent that was never observed after Qigong treatment. Aerobic training exerts a significant impact on the ability of moderately disabled PD patients to cope with exercise, although it does not improve their self-sufficiency and quality of life.


CHARMM: the biomolecular simulation program.
CHARMM (Chemistry at HARvard Molecular Mechanics) is a highly versatile and widely used molecular simulation program. It has been developed over the last three decades with a primary focus on molecules of biological interest, including proteins, peptides, lipids, nucleic acids, carbohydrates, and small molecule ligands, as they occur in solution, crystals, and membrane environments. For the study of such systems, the program provides a large suite of computational tools that include numerous conformational and path sampling methods, free energy estimators, molecular minimization, dynamics, and analysis techniques, and model-building capabilities. The CHARMM program is applicable to problems involving a much broader class of many-particle systems. Calculations with CHARMM can be performed using a number of different energy functions and models, from mixed quantum mechanical-molecular mechanical force fields, to all-atom classical potential energy functions with explicit solvent and various boundary conditions, to implicit solvent and membrane models. The program has been ported to numerous platforms in both serial and parallel architectures. This article provides an overview of the program as it exists today with an emphasis on developments since the publication of the original CHARMM article in 1983.


Pre- and postcopulatory sexual selection favor aggressive, young males in polyandrous groups of red junglefowl.
A challenge in evolutionary biology is to understand the operation of sexual selection on males in polyandrous groups, where sexual selection occurs before and after mating. Here, we combine fine-grained behavioral information (>41,000 interactions) with molecular parentage data to study sexual selection in replicated, age-structured groups of polyandrous red junglefowl, Gallus gallus. Male reproductive success was determined by the number of females mated (precopulatory sexual selection) and his paternity share, which was driven by the polyandry of his female partners (postcopulatory sexual selection). Pre- and postcopulatory components of male reproductive success covaried positively; males with high mating success also had high paternity share. Two male phenotypes affected male pre- and postcopulatory performance: average aggressiveness toward rival males and age. Aggressive males mated with more females and more often with individual females, resulting in higher sexual exclusivity. Similarly, younger males mated with more females and more often with individual females, suffering less intense sperm competition than older males. Older males had a lower paternity share even allowing for their limited sexual exclusivity, indicating they may produce less competitive ejaculates. These results show that-in these populations-postcopulatory sexual selection reinforces precopulatory sexual selection, consistently promoting younger and more aggressive males.


Persistent hamathecial threads in the Nectriaceae, Hypocreales: Thyronectria revisited and re-instated.
Based on type studies and freshly collected material we here re-instate the genus Thyronectria (Nectriaceae, Hypocreales). Species of this genus were recently for the most part classified in the genera Pleonectria (Nectriaceae) or Mattirolia (Thyridiaceae), because Thyronectria and other genera had been identified as members of the Thyridiaceae due to the presence of paraphyses. Molecular phylogenies based on several markers (act, ITS, LSU rDNA, rpb1, rpb2, tef1, tub) revealed that the Nectriaceae contain members whose ascomata are characterised by long, more or less persistent, apical paraphyses. All of these belong to a single genus, Thyronectria, which thus has representatives with hyaline, rosy, green or even dark brown and sometimes distoseptate ascospores. The type species of Thyronectria, T. rhodochlora, syn. T. patavina, syn. T. pyrrhochlora is re-described and illustrated. Within the Nectriaceae persistent, apical paraphyses are common in Thyronectria and rarely also occur in Nectria. The genus Mattirolia is revised and merged with Thyronectria and also Thyronectroidea is regarded as a synonym of Thyronectria. The three new species T. asturiensis, T. caudata and T. obscura are added to the genus. Species recently described in Pleonectria as well as some species of Mattirolia are combined in the genus, and a key to Thyronectria is provided. Five species are epitypified. The type species of the genus Thyridium (Thyridiaceae), T. vestitum, is included in phylogenetic analyses to illustrate the phylogenetic distance of Thyronectria from the Thyridiaceae.


[From corpsman to rescuer, the missed nurse's ideal towards the end of italian Risorgimento].
Military corps have always been supported by healthcare providers who took care of the injured and sick soldiers. Traditionally the military nurse's figure has never been fully appreciated to the point that, even nowadays, it is still searching for its own identity. The aim of the study is to describe the military nurse's role from the Second War for Italian Independence (1859) to the conquest of Rome (1870). Historical investigation. Secondary sources were consulted to carry out a geopolitical and historical contextualization of the reference period, while to trace the military nurse's evolution the team referred to primary sources. All the mentioned sources were analyzed according to Chabod's methodology (2012). The military nurse was born as a corpsman. The Risorgimento wars contributed to highlight the major contribution that nurses could give to military healthcare. In 1863 the Ministry of War elaborated a new and innovative profile for military nurses which foresaw particular physical and moral requirements, but also specific competences. Even though a new ideal of nurse was created, the Military Nurses Corp did not experience any changes: the nurses' activities remained transporting the injured and assisting doctors. Primary sources analysis highlighted an important attention towards the military nurse's role and education after national unity, although the gap shown by the military healthcare service during the Risorgimento battles was evident. Such condition facilitated the development of the first rescue committees which became, at a later time, the International Red Cross. The committees popularity allowed the volunteers to gain higher fame and social prestige than military nurses.


The ubiquitin-editing enzyme A20 restricts nucleotide-binding oligomerization domain containing 2-triggered signals.
Muramyl dipeptide (MDP), a product of bacterial cell-wall peptidoglycan, activates innate immune cells by stimulating nucleotide-binding oligomerization domain containing 2 (NOD2) -dependent activation of the transcription factor NFkappaB and transcription of proinflammatory genes. A20 is a ubiquitin-modifying enzyme that restricts tumor necrosis factor (TNF) receptor and Toll-like receptor (TLR) -induced signals. We now show that MDP induces ubiquitylation of receptor- interacting protein 2 (RIP2) in primary macrophages. A20-deficient cells exhibit dramatically amplified responses to MDP, including increased RIP2 ubiquitylation, prolonged NFkappaB signaling, and increased production of proinflammatory cytokines. In addition, in vivo responses to MDP are exaggerated in A20-deficient mice and in chimeric mice bearing A20-deficient hematopoietic cells. These exaggerated responses occur independently of the TLR adaptors MyD88 and TRIF as well as TNF signals. These findings indicate that A20 directly restricts NOD2 induced signals in vitro and in vivo, and provide new insights into how these signals are physiologically restricted.


Androgen receptor modulation following combination exposure to brominated flame-retardants.
Endocrine disrupting compounds can interfere with androgen receptor (AR) signaling and disrupt steroidogenesis leading to reproductive failure. The brominated flame-retardant (BFR) 1, 2-dibromo-4-(1, 2-dibromoethyl) cyclohexane (TBECH), is an agonist to human, chicken and zebrafish AR. Recently another group of alternative BFRs, allyl 2, 4, 6-tribromophenyl ether (ATE), and 2, 3-dibromopropyl 2, 4, 6-tribromophenyl ether (DPTE) along with its metabolite 2-bromoallyl 2, 4, 6-tribromophenyl ether (BATE) were identified as potent human AR antagonists. These alternative BFRs are present in the environment. The aim of the present study was to determine the effect of mixed exposures to the AR agonist and the AR antagonists at environmentally relevant concentrations. In vitro reporter luciferase assay showed that the AR antagonists, when present at concentration higher than TBECH, were able to inhibit TBECH-mediated AR activity. These AR antagonists also promoted AR nuclear translocation. In vitro gene expression analysis in the non-tumorigenic human prostate epithelial cell RWPE1 showed that TBECH induced AR target genes whereas DPTE repressed these genes. Further analysis of steroidogenic genes showed that TBECH up-regulated most of the genes while DPTE down-regulated the same genes. The results indicate that when TBECH and DPTE are present together they will antagonize each other, thereby reducing their individual effects.


Competition at silent synapses in reinnervated skeletal muscle.
Synaptic connections are made and broken in an activity-dependent manner in diverse regions of the nervous system. However, whether activity is strictly necessary for synapse elimination has not been resolved directly. Here we report that synaptic terminals occupying motor endplates made electrically silent by tetrodotoxin and alpha-bungarotoxin block were frequently displaced by regenerating axons that were also both inactive and synaptically ineffective. Thus, neither evoked nor spontaneous activation of acetylcholine receptors is required for competitive reoccupation of neuromuscular synaptic sites by regenerating motor axons.


Skin involvement and breast cancer: are T4b lesions of all sizes created equal?
Nonmetastatic, noninflammatory, invasive breast cancers with skin involvement (SI) are classified as T4b, regardless of size. This study evaluated disease-specific survival (DSS) to determine whether size should be considered for these lesions rather than grouping them all into stage III. Surveillance, Epidemiology, and End Results data linked to Medicare claims were reviewed. Skin involved and non-SI tumors were reclassified using the American Joint Committee on Cancer, 7(th) edition groupings using tumor size and nodal involvement alone without considering SI (neostage). Disease-specific survival was adjusted for demographics, histology, and treatment using competing risk methods with propensity score-based weighting and bootstrap standard errors. Among 924 SI patients diagnosed between 1992 and 2005, tumors were 0.1 to 2.0 cm, 2.1 to 5.0 cm, and >5.0 cm in 11.6%, 51.1%, and 37.3% of patients, respectively. There were no nodal metastases in 22.3%, 1 to 3 positive nodes in 31.7%, 4 to 9 positive in 28.6%, and ≥10 positive in 17.4% of patients. For SI patients, adjusted 5-year DSS was 95.8% (95% CI, 95.6-96.0) for neostage I, declining progressively to 36.4% (95% CI, 33.8-39.2) for neostage IIIC patients. Adjusted 5-year DSS for SI and non-SI tumors (n = 66,185) was similar for neostage I, IIA, and IIB, and markedly lower for IIIA and IIIC. Adjusted DSS for SI IIIA was similar to non-SI IIIC. Noninflammatory SI breast cancers have widely varied DSS that differs by tumor size and nodal involvement and therefore should not all be stage III. Skin involvement should be subordinate to T and N groupings to classify SI with non-SI lesions having similar prognoses.


Selective proteolysis of immunoglobulins by mouse macrophage elastase.
Mouse macrophage elastase, a metalloproteinase secreted by inflammatory macrophages, catalyzed the limited proteolysis of selected subclasses of mouse immunoglobulins, including monomeric IgG2a, IgG3, and some forms of IgG2b. Mouse IgG1 was resistant to elastase degradation; however, human IgG1 was degraded. IgG3 in immune complexes was cleaved in a manner similar to that of monomeric IgG3. Degradation by macrophage elastase was limited to the heavy chain, resulting in products that did not compete for binding to the macrophage Fc receptor. Macrophage elastase usually produced a pepsin-like rather than a papain-like pattern of proteolysis, resulting in the release of F(ab')2 and Fc' subfragments. This degradation of IgG differed from the papain-like cleavage of IgG by granulocyte elastase. Macrophage elastase degraded papain-generated Fc fragments of IgG2a into multiple fragments. Therefore, macrophage elastase at concentrations found in culture medium has the potential to regulate some aspects of cellular events associated with immunoglobulins.


Determinants of the use of coronary angiography and revascularization after thrombolysis for acute myocardial infarction.
Clinical trials and practice guidelines have identified clinical criteria for the use of coronary angiography and revascularization procedures after thrombolysis for acute myocardial infarction. The effect of these criteria on clinical practice has not been extensively evaluated. We used classification-and-regression-tree (CART) and logistic-regression models to study the patients in the first Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries trial, to identify the variables that best predicted the use of angiography and revascularization procedures after thrombolysis. Among the 21,772 U.S. patients in the trial, 71 percent underwent coronary angiography before discharge from the hospital. Of these, 58 percent underwent revascularization (73 percent receiving angioplasty). The CART model for the use of angiography showed that age was the variable most predictive of angiography; only 53 percent of patients at least 73 years of age underwent angiography, as compared with 76 percent of those under 73. Among the older patients, age was again the most predictive factor; among the younger patients, the availability of angioplasty was a more important predictor (67 percent of patients in hospitals without angioplasty facilities underwent angiography, as compared with 83 percent in hospitals with such facilities). The next most important variable was recurrent ischemia, which was more predictive at hospitals without angioplasty facilities than at those with them. Both statistical models identified coronary anatomy as the most important predictor of the use and type of revascularization. More patients treated with thrombolysis underwent angiography and revascularization before discharge than might be expected. Younger age and the availability of the procedures appeared to be the major determinants of the use of coronary angiography, whereas coronary anatomy largely determined the use and type of revascularization. This process appeared to select low-risk patients for intervention rather than those at higher risk, who would be the most likely to benefit.


The lipid phosphatase-like protein PLPPR1 associates with RhoGDI1 to modulate RhoA activation in response to axon growth inhibitory molecules.
Phospholipid Phosphatase-Related Protein Type 1 (PLPPR1) is a member of a family of lipid phosphatase related proteins, integral membrane proteins characterized by six transmembrane domains. This family of proteins is enriched in the brain and recent data indicate potential pleiotropic functions in several different contexts. An inherent ability of this family of proteins is to induce morphological changes, and we have previously reported that members of this family interact with each other and may function co-operatively. However, the function of PLPPR1 is not yet understood. Here we show that the expression of PLPPR1 reduces the inhibition of neurite outgrowth of cultured mouse hippocampal neurons by chondroitin sulfate proteoglycans and the retraction of neurites of Neuro-2a cells by lysophosphatidic acid (LPA). Further, we show that PLPPR1 reduces the activation of Ras homolog family member A (RhoA) by LPA in Neuro-2a cells, and that this is because of an association of PLPPR1with the Rho-specific guanine nucleotide dissociation inhibitor (RhoGDI1). These results establish a novel signaling pathway for the PLPPR1 protein.


The role of the endoplasmic reticulum Ca2+ store in the plasticity of central neurons.
The smooth endoplasmic reticulum (SER) is a well-characterized buffer and source of Ca2+ in both axonal and dendritic compartments of neurons. Ca2+ release from the SER can be evoked by stimulation of the ryanodine receptor or the inositol (1,4,5)-trisphosphate [Ins(1,4,5)P3] receptor. Both receptors can couple to the activation of neurotransmitter-gated receptors and voltage-gated Ca2+ channels on the plasma membrane, thus enabling the SER to discriminate between different types of neuronal activity. In axonal terminals, Ca2+-induced Ca2+ release (CICR) mediates spontaneous, evoked and facilitated neurotransmission. Store release might also regulate the mobilization and recycling of synaptic vesicles. In the dendritic compartment, the distribution of Ins(1,4,5)P3 receptors and ryanodine receptors influences the intracellular encoding of neuronal activity. Thus, the functionality of the Ca2+ store can affect both the polarity and the spatial extent of Ca2+-dependent shifts in synaptic efficacy. In hippocampal neurons, for example, CICR in the spine heads underlies homosynaptic plasticity, whereas heterosynaptic plasticity is mediated by Ins(1,4,5)P3-dependent Ca2+ signalling. Purkinje neurons primarily express Ins(1,4,5)P3 receptors in the spine heads, and long-term depression of synaptic efficacy is crucially dependent on Ins(1,4,5)P3.


Super-resolution microscopy can identify specific protein distribution patterns in platelets incubated with cancer cells.
Protein contents in platelets are frequently changed upon tumor development and metastasis. However, how cancer cells can influence protein-selective redistribution and release within platelets, thereby promoting tumor development, remains largely elusive. With fluorescence-based super-resolution stimulated emission depletion (STED) imaging we reveal how specific proteins, implicated in tumor progression and metastasis, re-distribute within platelets, when subject to soluble activators (thrombin, adenosine diphosphate and thromboxane A2), and when incubated with cancer (MCF-7, MDA-MB-231, EFO21) or non-cancer cells (184A1, MCF10A). Upon cancer cell incubation, the cell-adhesion protein P-selectin was found to re-distribute into circular nano-structures, consistent with accumulation into the membrane of protein-storing alpha-granules within the platelets. These changes were to a significantly lesser extent, if at all, found in platelets incubated with normal cells, or in platelets subject to soluble platelet activators. From these patterns, we developed a classification procedure, whereby platelets exposed to cancer cells, to non-cancer cells, soluble activators, as well as non-activated platelets all could be identified in an automatic, objective manner. We demonstrate that STED imaging, in contrast to electron and confocal microscopy, has the necessary spatial resolution and labelling efficiency to identify protein distribution patterns in platelets and can resolve how they specifically change upon different activations. Combined with image analyses of specific protein distribution patterns within the platelets, STED imaging can thus have a role in future platelet-based cancer diagnostics and therapeutic monitoring. The presented approach can also bring further clarity into fundamental mechanisms for cancer cell-platelet interactions, and into non-contact cell-to-cell interactions in general.


Guidelines and good clinical practice recommendations for Contrast Enhanced Ultrasound (CEUS) in the liver - update 2012: A WFUMB-EFSUMB initiative in cooperation with representatives of AFSUMB, AIUM, ASUM, FLAUS and ICUS.
Initially, a set of guidelines for the use of ultrasound contrast agents was published in 2004 dealing only with liver applications. A second edition of the guidelines in 2008 reflected changes in the available contrast agents and updated the guidelines for the liver, as well as implementing some non-liver applications. Time has moved on, and the need for international guidelines on the use of CEUS in the liver has become apparent. The present document describes the third iteration of recommendations for the hepatic use of contrast enhanced ultrasound (CEUS) using contrast specific imaging techniques. This joint WFUMB-EFSUMB initiative has implicated experts from major leading ultrasound societies worldwide. These liver CEUS guidelines are simultaneously published in the official journals of both organizing federations (i.e., Ultrasound in Medicine and Biology for WFUMB and Ultraschall in der Medizin/European Journal of Ultrasound for EFSUMB). These guidelines and recommendations provide general advice on the use of all currently clinically available ultrasound contrast agents (UCA). They are intended to create standard protocols for the use and administration of UCA in liver applications on an international basis and improve the management of patients worldwide.


Exploration of underwater life with an acoustically controlled soft robotic fish.
Closeup exploration of underwater life requires new forms of interaction, using biomimetic creatures that are capable of agile swimming maneuvers, equipped with cameras, and supported by remote human operation. Current robotic prototypes do not provide adequate platforms for studying marine life in their natural habitats. This work presents the design, fabrication, control, and oceanic testing of a soft robotic fish that can swim in three dimensions to continuously record the aquatic life it is following or engaging. Using a miniaturized acoustic communication module, a diver can direct the fish by sending commands such as speed, turning angle, and dynamic vertical diving. This work builds on previous generations of robotic fish that were restricted to one plane in shallow water and lacked remote control. Experimental results gathered from tests along coral reefs in the Pacific Ocean show that the robotic fish can successfully navigate around aquatic life at depths ranging from 0 to 18 meters. Furthermore, our robotic fish exhibits a lifelike undulating tail motion enabled by a soft robotic actuator design that can potentially facilitate a more natural integration into the ocean environment. We believe that our study advances beyond what is currently achievable using traditional thruster-based and tethered autonomous underwater vehicles, demonstrating methods that can be used in the future for studying the interactions of aquatic life and ocean dynamics.


Comprehending the lack of access to maternal and neonatal emergency care: Designing solutions based on a space-time approach.
The objective of this study was to better understand how the lack of emergency child and obstetric care can be related to maternal and neonatal mortality levels. We performed spatiotemporal geospatial analyses using data from Brazilian municipalities. An emergency service accessibility index was derived using the two-step floating catchment area (2SFCA) for 951 hospitals. Mortality data from 2000 to 2015 was used to characterize space-time trends. The data was overlapped using a spatial clusters analysis to identify regions with lack of emergency access and high mortality trends. From 2000 to 2015 Brazil the overall neonatal mortality rate varied from 11,42 to 11,71 by 1000 live births. The maternal mortality presented a slightly decrease from 2,98 to 2,88 by 100 thousand inhabitants. For neonatal mortality the Northeast and North regions presented the highest percentage of up trending. For maternal mortality the North region exhibited the higher volume of up trending. The accessibility index obtained highlighted large portions of the rural areas of the country without any coverage of obstetric or neonatal beds. The analyses highlighted regions with problems of mortality and access to maternal and newborn emergency services. This sequence of steps can be applied to other low and medium income countries as health situation analysis tool. Low and middle income countries have greater disparities in access to emergency child and obstetric care. There is a lack of approaches capable to support analysis considering a spatiotemporal perspective for emergency care. Studies using Geographic Information System analysis for maternal and child care, are increasing in frequency. This approach can identify emergency child and obstetric care saturated or deprived regions. The sequence of steps designed here can help researchers, and policy makers to better design strategies aiming to improve emergency child and obstetric care.


Isomerization of methoxy radical in the troposphere: competition between acidic, neutral and basic catalysts.
A comprehensive investigation of the roles of acidic, neutral and basic catalysts in isomerization of methoxy radical in the troposphere has been carried out by quantum chemical calculations at the MP2 and CCSD(T) levels of theory. The effect of basic catalysts, namely ammonia and an ammonia-water complex, on the isomerization process has been studied for the very first time. In terms of rate coefficients ammonia was found to be a better catalyst than a water monomer whereas the ammonia-water complex was found to be more efficient over a water dimer but marginally less efficient than formic acid. Based on the effective rate constants under various tropospheric conditions, it was found that at 0 km altitude water dimers and ammonia-water complexes could compete with acid catalysts but at higher altitudes the acid catalysts would dominate their neutral and basic counterparts by a long distance.


Use of the Agilent 2100 bioanalyzer for rapid and reproducible molecular typing of Streptococcus pneumoniae.
Restriction fragment length polymorphism (RFLP) analysis is an economic and fast technique for molecular typing but has the drawback of difficulties in accurately sizing DNA fragments and comparing banding patterns on agarose gels. We aimed to improve RFLP for typing of the important human pathogen Streptococcus pneumoniae and to compare the results with the commonly used typing techniques of pulsed-field gel electrophoresis and multilocus sequence typing. We designed primers to amplify a noncoding region adjacent to the pneumolysin gene. The PCR product was digested separately with six restriction endonucleases, and the DNA fragments were analyzed using an Agilent 2100 bioanalyzer for accurate sizing. The combined RFLP results for all enzymes allowed us to assign each of the 47 clinical isolates of S. pneumoniae tested to one of 33 RFLP types. RFLP analyzed using the bioanalyzer allowed discrimination between strains similar to that obtained by the more commonly used techniques of pulsed-field gel electrophoresis, which discriminated between 34 types, and multilocus sequence typing, which discriminated between 35 types, but more quickly and with less expense. RFLP of a noncoding region using the Agilent 2100 bioanalyzer could be a useful addition to the molecular typing techniques in current use for S. pneumoniae, especially as a first screen of a local population.


Control of early Theiler's murine encephalomyelitis virus replication in macrophages by interleukin-6 occurs in conjunction with STAT1 activation and nitric oxide production.
During Theiler's murine encephalomyelitis virus (TMEV) infection of macrophages, it is thought that high interleukin-6 (IL-6) levels contribute to the demyelinating disease found in chronically infected SJL/J mice but absent in B10.S mice capable of clearing the infection. Therefore, IL-6 expression was measured in TMEV-susceptible SJL/J and TMEV-resistant B10.S macrophages during their infection with TMEV DA strain or responses to lipopolysaccharide (LPS) or poly(I · C). Unexpectedly, IL-6 production was greater in B10.S macrophages than SJL/J macrophages during the first 24 h after stimulation with TMEV, LPS, or poly(I · C). Further experiments showed that in B10.S, SJL/J, and RAW264.7 macrophage cells, IL-6 expression was dependent on extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) and enhanced by exogenous IL-12. In SJL/J and RAW264.7 macrophages, exogenous IL-6 resulted in decreased TMEV replication, earlier activation of STAT1 and STAT3, production of nitric oxide, and earlier upregulation of several antiviral genes downstream of STAT1. However, neither inhibition of IL-6-induced nitric oxide nor knockdown of STAT1 diminished the early antiviral effect of exogenous IL-6. In addition, neutralization of endogenous IL-6 from SJL/J macrophages with Fab antibodies did not exacerbate early TMEV infection. Therefore, endogenous IL-6 expression after TMEV infection is dependent on ERK MAPK, enhanced by IL-12, but too slow to decrease viral replication during early infection. In contrast, exogenous IL-6 enhances macrophage control of TMEV infection through preemptive antiviral nitric oxide production and antiviral STAT1 activation. These results indicate that immediate-early production of IL-6 could protect macrophages from TMEV infection.


Distinct Corticospinal and Reticulospinal Contributions to Voluntary Control of Elbow Flexor and Extensor Muscles in Humans with Tetraplegia.
Humans with cervical spinal cord injury (SCI) often recover voluntary control of elbow flexors and, to a much lesser extent, elbow extensor muscles. The neural mechanisms underlying this asymmetrical recovery remain unknown. Anatomical and physiological evidence in animals and humans indicates that corticospinal and reticulospinal pathways differentially control elbow flexor and extensor motoneurons; therefore, it is possible that reorganization in these pathways contributes to the asymmetrical recovery of elbow muscles after SCI. To test this hypothesis, we examined motor-evoked potentials (MEPs) elicited by transcranial magnetic stimulation over the arm representation of the primary motor cortex, maximal voluntary contractions, the StartReact response (a shortening in reaction time evoked by a startling stimulus), and the effect of an acoustic startle cue on MEPs elicited by cervicomedullary stimulation (CMEPs) on biceps and triceps brachii in males and females with and without chronic cervical incomplete SCI. We found that SCI participants showed similar MEPs and maximal voluntary contractions in biceps but smaller responses in triceps compared with controls, suggesting reduced corticospinal inputs to elbow extensors. The StartReact and CMEP facilitation was larger in biceps but similar to controls in triceps, suggesting enhanced reticulospinal inputs to elbow flexors. These findings support the hypothesis that the recovery of biceps after cervical SCI results, at least in part, from increased reticulospinal inputs and that the lack of these extra inputs combined with the loss of corticospinal drive contribute to the pronounced weakness found in triceps.SIGNIFICANCE STATEMENT Although a number of individuals with cervical incomplete spinal cord injury show limited functional recovery of elbow extensors compared with elbow flexor muscles, to date, the neural mechanisms underlying this asymmetrical recovery remain unknown. Here, we provide for the first time evidence for increased reticulospinal inputs to biceps but not triceps brachii and loss of corticospinal drive to triceps brachii in humans with tetraplegia. We propose that this reorganization in descending control contributes to the asymmetrical recovery between elbow flexor and extensor muscles after cervical spinal cord injury.


[Interculturality in health].
Interculturality in health is a recent concept that develops in response to claims of indigenous peoples for their right to their cultural identity, but also correspond to a global trend of incorporating the "right to the difference", which distinguishes and promotes coexistence between different cultural groups in the same territory. The article discusses, from a historical perspective, the original mismatch between Native American populations and European conquerors that marked out their access to health services, and discussed some of the many current issues related to, as the identification of indigenous people and the relationship between human rights and interculturality, to finally present a review of the genesis of the concept of interculturalism in health and their complexity reviewing the concept of cultural syndrome and his adaptation to scientific medicine.


Severe E-Cigarette, or Vaping, Product Use Associated Lung Injury Requiring Venovenous Extracorporeal Membrane Oxygenation.
To report a severe case of e-cigarette or vaping product use-associated lung injury with complex course requiring venovenous extracorporeal membrane oxygenation. Case report. PICU in an academic medical center. A 16-year-old girl presenting with gastrointestinal and respiratory symptoms was admitted to our PICU after having progressive respiratory failure and bilateral pulmonary ground-glass opacities on chest CT. Venovenous extracorporeal membrane oxygenation MEASUREMENTS AND MAIN RESULTS:: After extensive infectious workup was unrevealing, she reported a history of vaping e-cigarette containing either nicotine or delta-9-tetrahydrocannabinol oil prior to symptom onset. She was given a presumptive diagnosis of e-cigarette or vaping product use-associated lung injury. The PICU team in consultation with pulmonology and medical toxicology started high-dose IV methylprednisolone 1 mg/kg bid. Despite initial improvements, she continued to require positive pressure ventilation and developed pneumomediastinum with progression to tension pneumothoraces and a persistent air leak. Unable to maintain her oxygenation, she was placed on venovenous extracorporeal membrane oxygenation for a prolonged course and had a tracheostomy placement. The clinical course, severity, and range of interventions in affected patients around the country have varied widely. Respiratory symptoms have been the most severe, but the constellation of symptoms in e-cigarette or vaping product use-associated lung injury include constitutional symptoms (fevers, weight-loss) and gastrointestinal symptoms (nausea, vomiting, diarrhea). In many cases, steroid use led to rapid clinical improvements. However, other cases with severe illness, like our patient, necessitated high-dose IV steroids, intubation, and venovenous extracorporeal membrane oxygenation. The underlying etiology and pathophysiology of e-cigarette or vaping product use-associated lung injury remains unknown. The Centers for Disease Control and Prevention in conjunction with state/local health departments and the Food and Drug Administration is actively investigating the outbreak. Clinicians need to be aware of the current outbreak of e-cigarette or vaping product use-associated lung injury and ask about vaping in patients presenting with gastrointestinal and respiratory symptoms. Treatment options are anecdotal and necessitate a multidisciplinary approach.


The relationship between exercise,nutrition and type 2 diabetes.
Type 2 diabetes mellitus and its precursor, insulin resistance, are metabolic disease states characterized by impaired regulation in the delivery, transport, and/or storage of energy substrates (primarily carbohydrate- and fat-based fuels). A hallmark feature of patients with type 2 diabetes is prolonged periods of hyperglycemia due to a decreased responsiveness of metabolically active peripheral tissues to the actions of insulin (i.e., metabolic inflexibility). Accordingly, efforts to modify skeletal muscle substrate handling in type 2 diabetes patients so that the capacity for fat oxidation and metabolic flexibility is improved should be a primary goal for the treatment of these disorders. Two potent interventions for improving whole-body glucose homeostasis are exercise and diet. A single bout of either resistance or endurance exercise reduces the prevalence and duration of hyperglycemic excursions in patients with type 2 diabetes, an effect lasting well into the next day. With regard to diet, the carbohydrate content of a meal and the glycemic index (GI) of the carbohydrate consumed are both major determinants of the postprandial glycemic response. Diets containing high-GI carbohydrates have been shown to be independent risk factors for type 2 diabetes onset, while in obese insulin-resistant individuals, low-GI diets are effective for inducing both weight loss and improving insulin action and glucose tolerance. The implementation of physical activity and dietary modifications are effective low-cost treatment options for controlling hyperglycemic episodes in patients with type 2 diabetes.


Suicide and suicide attempts in adolescents.
Suicide is the third-leading cause of death for adolescents 15 to 19 years old. Pediatricians can take steps to help reduce the incidence of adolescent suicide by screening for depression and suicidal ideation and behavior. This report updates the previous statement of the American Academy of Pediatrics and is intended to assist the pediatrician in the identification and management of the adolescent at risk of suicide. The extent to which pediatricians provide appropriate care for suicidal adolescents depends on their knowledge, skill, comfort with the topic, and ready access to appropriate community resources. All teenagers with suicidal thoughts or behaviors should know that their pleas for assistance are heard and that pediatricians are willing to serve as advocates to help resolve the crisis.


Xenopus Mcm10 is a CDK-substrate required for replication fork stability.
During S phase, following activation of the S phase CDKs and the DBF4-dependent kinases (DDK), double hexamers of Mcm2-7 at licensed replication origins are activated to form the core replicative helicase. Mcm10 is one of several proteins that have been implicated from work in yeasts to play a role in forming a mature replisome during the initiation process. Mcm10 has also been proposed to play a role in promoting replisome stability after initiation has taken place. The role of Mcm10 is particularly unclear in metazoans, where conflicting data has been presented. Here, we investigate the role and regulation of Mcm10 in Xenopus egg extracts. We show that Xenopus Mcm10 is recruited to chromatin late in the process of replication initiation and this requires prior action of DDKs and CDKs. We also provide evidence that Mcm10 is a CDK substrate but does not need to be phosphorylated in order to associate with chromatin. We show that in extracts depleted of more than 99% of Mcm10, the bulk of DNA replication still occurs, suggesting that Mcm10 is not required for the process of replication initiation. However, in extracts depleted of Mcm10, the replication fork elongation rate is reduced. Furthermore, the absence of Mcm10 or its phosphorylation by CDK results in instability of replisome proteins on DNA, which is particularly important under conditions of replication stress.


Caging of Cl atoms from photodissociation of CF2Cl2 in clusters.
We investigate the photodissociation dynamics of freon CF2Cl2 by velocity map imaging at 193 nm. The Cl fragment images are recorded in various expansion regimes corresponding to isolated molecules and molecules in rare gas clusters. The molecular kinetic energy distributions are dominated by a peak at Ekin(Cl) ≈ 0.97 eV corresponding to the direct C-Cl bond dissociation but they also reveal features at lower kinetic energies. Possible mechanisms leading to these slow Cl atoms are discussed. The photodissociation in clusters is investigated in two regimes: (i) small Ar clusters with the CF2Cl2 molecule embedded in approximately one solvation Ar layer; (ii) large Xen, n¯ ≈ 100-500, clusters with embedded CF2Cl2 molecules. In the former clusters we observe caging yielding the Cl fragments with zero kinetic energy and direct exit resulting in fragments with the kinetic energies corresponding to the fragments from isolated molecules. In the latter case (ii) only the caged Cl fragments are observed.


Dual-Responsive pH and Temperature Sensitive Nanoparticles Based on Methacrylic Acid and Di(ethylene glycol) Methyl Ether Methacrylate for the Triggered Release of Drugs.
A series of thermo-and pH-responsive poly(methyl methacrylate)-block-poly[methacrylic acid-co-di(ethylene glycol) methyl ether methacrylate] PMMA-b-P[MAA-co-DEGMA] block copolymers were synthesized by RAFT polymerization and self-assembled into micelles. The molar ratio of MAA was altered from 0-12% in order to modulate the lower critical solution temperature (LCST) of PDEGMA. The release of the drug albendazole from the micelle was strongly dependent on the temperature and the LCST value of the polymer. Systems below the LCST released the drug slowly while increasing the temperature above the LCST or decreasing the pH value to 5 resulted in the burst-like release of the drug. ABZ delivered in this pH-responsive drug carrier had a higher toxicity than the free drug or the drug delivered in a non-responsive drug carrier.


Protein-binding elements in the promoter region of the mouse p53 gene.
p53 is a cellular protein whose expression plays a crucial role in the regulation of cell proliferation and of neoplastic processes. p53 mRNA levels in mouse fibroblasts can be elevated in response to TPA and to serum stimulation. The promoter region of the p53 gene contains a conserved element which is highly homologous to the consensus AP1 binding site (7/8 matching bases). This AP1-like site, denoted the PF1 site, confers upon a heterologous promoter ability to respond to elevated expression of c-jun. Furthermore, the PF1 site binds protein(s) in a specific and serum-induced manner. Unexpectedly, this factor is most probably not AP1, as evident from the inability of an authentic AP1 site to compete the binding efficiently, as well as from the failure of purified AP1 to bind to the PF1 site. Hence, PF1 may be a novel AP1-related transcription factor. In addition, the 5' region of the p53 gene also contains an NF1 binding site, whose location suggests a possible regulatory role.


Procarbazine-free OEPA-COPDAC chemotherapy in boys and standard OPPA-COPP in girls have comparable effectiveness in pediatric Hodgkin's lymphoma: the GPOH-HD-2002 study.
Vincristine, etoposide, prednisone, and doxorubicin (OEPA)-cyclophosphamide, vincristine, prednisone, and dacarbazine (COPDAC) is derived from standard vincristine, procarbazine, prednisone, and doxorubicin (OPPA)-cyclophosphamide, vincristine, procarbazine, and prednisone (COPP) chemotherapy by replacing procarbazine with etoposide and dacarbazine for a potentially less gonadotoxic regimen for boys with Hodgkin's lymphoma (HL). Five hundred seventy-three pediatric patients with classical HL were enrolled onto the German Society of Pediatric Oncology and Hematology-Hodgkin's Disease (GPOH-HD) -2002 study between November 2002 and December 2005. Boys received two courses of OEPA and girls received two courses of OPPA for induction. Treatment group (TG) -2 (intermediate stages) and TG-3 (advanced stages) patients received further two or four cycles COPP (girls) or COPDAC (boys), respectively. After chemotherapy all patients received involved-field irradiation with 19.8 Gy, except for patients with early-stage disease (TG-1) in complete remission. Five hundred seventy-three patients (287 males and 286 females) were less than 18 years old and fulfilled all inclusion criteria; 195 patients (34.0%) were allocated to TG-1, 139 (24.3%) were allocated to TG-2, and 239 (41.7%) were allocated to TG-3. Toxicity of OEPA-COPDAC was tolerable overall. Hematotoxicity was more pronounced with OEPA than OPPA, whereas it was less pronounced with COPDAC compared with COPP. The median observation time was 58.6 months. Overall survival and event-free survival (EFS) rates (+/- SE) at 5 years were 97.4% +/- 0.7% and 89.0% +/- 1.4%, respectively. In TG-1, overall EFS was 92.0% +/- 2.0%. EFS of patients without irradiation (93.2% +/- 3.3%) was similar to that of irradiated patients (91.7% +/- 2.5%), confirming results of the previous GPOH-HD-95 study. In TG-2+3, EFS did not significantly differ between boys and girls (90.2% +/- 2.3 v 84.7% +/- 2.7, respectively; P = .12). In TG-2+3, results in boys and girls are superimposable. OPPA-COPP and OEPA-COPDAC seem to be exchangeable regimens in intermediate- and advanced-stage classical HL in pediatric patients.


Protective effect of heat shock protein 70 against oxidative stresses in human corneal fibroblasts.
We evaluated DNA protection effect of heat shock protein (HSP) against cytotoxic effects of exogenous nitric oxide (NO) and reactive oxygen intermediate (ROI). Cultured human corneal fibroblasts were divided into 4 groups. Control (Group I) was not exposed to a sub-lethal heat treatment. Other 3 groups were exposed to 43 degrees C for 1 hr, then incubated at 37 degrees C during different duration (1, 6, 24 hr, Group II, III, IV, respectively). Expression pattern of HSP 70 was analyzed by Western blot. Cell viability was measured by MTT assay and the relationship between HSP 70 expression and DNA damage was examined by terminal deoxyribonucleotidyl transferase mediated dUTP-digoxigenin nick and labeling (TUNEL) stain and single cell gel electrophoresis. Expression pattern of HSP 70 was dependent on recovery times. Cell viability following heat treatment was significantly increased and the TUNEL positive cell number was decreased at 6 hr. In single cell gel electrophoresis, tail moments were increased in a dose-dependent manner by SNAP and X/XO. Following heat treatment, tail moments showed decreased significantly at 6 hr. These results suggest that induction of HSP 70 by sub-lethal heat treatment is closely related with cytoprotective effects against oxidative stresses in human corneal fibroblasts.


Cancer stem cells: small subpopulation or evolving fraction?
Central to the debate about cancer stem cells in solid tumors is the proportion of cells that can initiate, propagate, and re-initiate tumors. An initially assumed minor subpopulation is confronted with recent data suggesting as many as 30% of primary tumor cells have stem cell characteristics. This review discusses quantitative modeling studies that augment our understanding of stem and non-stem cancer cell interactions during tumor progression and the resulting fraction of cancer stem cells. A discussion of how these findings can be carefully evaluated in novel, integrated interdisciplinary studies is offered.


Targeted Drug Delivery and Image-Guided Therapy of Heterogeneous Ovarian Cancer Using HER2-Targeted Theranostic Nanoparticles.
Cancer heterogeneity and drug resistance limit the efficacy of cancer therapy. To address this issue, we have developed an integrated treatment protocol for effective treatment of heterogeneous ovarian cancer. Methods: An amphiphilic polymer coated magnetic iron oxide nanoparticle was conjugated with near infrared dye labeled HER2 affibody and chemotherapy drug cisplatin. The effects of the theranostic nanoparticle on targeted drug delivery, therapeutic efficacy, non-invasive magnetic resonance image (MRI)-guided therapy, and optical imaging detection of therapy resistant tumors were examined in an orthotopic human ovarian cancer xenograft model with highly heterogeneous levels of HER2 expression. Results: We found that systemic delivery of HER2-targeted magnetic iron oxide nanoparticles carrying cisplatin significantly inhibited the growth of primary tumor and peritoneal and lung metastases in the ovarian cancer xenograft model in nude mice. Differential delivery of theranostic nanoparticles into individual tumors with heterogeneous levels of HER2 expression and various responses to therapy were detectable by MRI. We further found a stronger therapeutic response in metastatic tumors compared to primary tumors, likely due to a higher level of HER2 expression and a larger number of proliferating cells in metastatic tumor cells. Relatively long-time retention of iron oxide nanoparticles in tumor tissues allowed interrogating the relationship between nanoparticle drug delivery and the presence of resistant residual tumors by in vivo molecular imaging and histological analysis of the tumor tissues. Following therapy, most of the remaining tumors were small, primary tumors that had low levels of HER2 expression and nanoparticle drug accumulation, thereby explaining their lack of therapeutic response. However, a few residual tumors had HER2-expressing tumor cells and detectable nanoparticle drug delivery but failed to respond, suggesting additional intrinsic resistant mechanisms. Nanoparticle retention in the small residual tumors, nevertheless, produced optical signals for detection by spectroscopic imaging. Conclusion: The inability to completely excise peritoneal metastatic tumors by debulking surgery as well as resistance to chemotherapy are the major clinical challenges for ovarian cancer treatment. This targeted cancer therapy has the potential for the development of effective treatment for metastatic ovarian cancer.


An Improved 2-Dimensional Gel Electrophoresis Method for Resolving Human Erythrocyte Membrane Proteins.
The 2-dimensional gel electrophoresis (2-DE) technique is widely used for the analysis of complex protein mixtures extracted from biological samples. It is one of the most commonly used analytical techniques in proteomics to study qualitative and quantitative protein changes between different states of a cell or an organism (eg, healthy and diseased), conditionally expressed proteins, posttranslational modifications, and so on. The 2-DE technique is used for its unparalleled ability to separate thousands of proteins simultaneously. The resolution of the proteins by 2-DE largely depends on the quality of sample prepared during protein extraction which increases results in terms of reproducibility and minimizes protein modifications that may result in artifactual spots on 2-DE gels. The buffer used for the extraction and solubilization of proteins influences the quality and reproducibility of the resolution of proteins on 2-DE gel. The purification by cleanup kit is another powerful process to prevent horizontal streaking which occurs during isoelectric focusing due to the presence of contaminants such as salts, lipids, nucleic acids, and detergents. Erythrocyte membrane proteins serve as prototypes for multifunctional proteins in various erythroid and nonerythroid cells. In this study, we therefore optimized the selected major conditions of 2-DE for resolving various proteins of human erythrocyte membrane. The modification included the optimization of conditions for sample preparation, cleanup of protein sample, isoelectric focusing, equilibration, and storage of immobilized pH gradient strips, which were further carefully examined to achieve optimum conditions for improving the quality of protein spots on 2-DE gels. The present improved 2-DE analysis method enabled better detection of protein spots with higher quality and reproducibility. Therefore, the conditions established in this study may be used for the 2-DE analysis of erythrocyte membrane proteins for different diseases, which may help to identify the proteins that may serve as markers for diagnostics as well as targets for development of new therapeutic potential.


Acute hypoxia occludes hTREK-1 modulation: re-evaluation of the potential role of tandem P domain K+ channels in central neuroprotection.
The human tandem P domain K+ channel hTREK-1 (KCNK2) is distributed widely through the CNS. Here, whole-cell patch clamp recordings were employed to investigate the effects of hypoxia on hTREK-1 channels stably expressed in human embryonic kidney cells. Acute hypoxia caused a rapid and reversible inhibition of whole-cell K+ current amplitudes; this was PO2 dependent with a maximal inhibition achieved at 60 mmHg and below. In accordance with previous studies, hTREK-1 current amplitudes were enhanced by arachidonic acid. This effect was concentration dependent, with maximal enhancement observed at a concentration of 10 microM. Membrane deformation by the crenator trinitrophenol (to mimic cell swelling) or the cup former chlorpromazine (to mimic cell shrinkage) caused robust activation and inhibition of currents, respectively. However, current augmentation by either arachidonic acid or trinitrophenol was completely prevented during hypoxia; conversely, hypoxia blunted the inhibitory action of chlorpromazine. The abilities of arachidonic acid to augment currents and of hypoxia to completely abrogate this effect were also observed in cell-attached patches. Our data indicate that hypoxia interacts with hTREK-1, and occludes its modulation by arachidonic acid and membrane deformation. These findings also suggest that the potential neuroprotective role of TREK channels, which has recently been proposed, requires reconsideration since hTREK-1 activation is unlikely when ambient PO2 is below 60 mmHg - a situation which normally pertains in the CNS even during systemic normoxia.


CO2 driven endotracheal tube cuff control in critically ill patients: A randomized controlled study.
To determine the safety and clinical efficacy of an innovative integrated airway system (AnapnoGuard™ 100 system) that continuously monitors and controls the cuff pressure (Pcuff), while facilitating the aspiration of subglottic secretions (SS). This was a prospective, single centre, open-label, randomized, controlled feasibility and safety trial. The primary endpoint of the study was the rate of device related adverse events (AE) and serious AE (SAE) as a result of using AnapnoGuard (AG) 100 during mechanical ventilation. Secondary endpoints were: (1) mechanical complications rate (2) ICU staff satisfaction; (3) VAP occurrence; (4) length of mechanical ventilation; (5) length of Intensive Care Unit stay and mortality; (6) volume of evacuated subglottic secretions. Sixty patients were randomized to be intubated with the AG endotracheal-tube (ETT) and connected to the AG 100 system allowing Pcuff adjustment and SS aspiration; or with an ETT combined with SS drainage and Pcuff controlled manually. No difference in adverse events rate was identified between the groups. The use of AG system was associated with a significantly higher incidence of Pcuff determinations in the safety range (97.3% vs. 71%; p<0.01) and a trend to a greater volume of aspirated SS secretions: (192.0[64-413] ml vs. 150[50-200], p = 0.19 (total)); (57.8[20-88.7] ml vs. 50[18.7-62] ml, p = 0.11 (daily)). No inter-group difference was detected using AG system vs. controls in terms of post-extubation throat pain level (0 [0-2] vs. 0 [0-3]; p = 0.7), hoarseness (42.9% vs. 75%; p = 0.55) and tracheal mucosa oedema (16.7% vs. 10%; p = 0.65). Patients enrolled in the AG group had a trend to reduced VAP risk of ventilator-associated pneumonia(VAP) (14.8% vs. 40%; p = 0.06), which were more frequently monomicrobial (25% vs. 70%; p = 0.03). No statistically significant difference was observed in duration of mechanical ventilation, ICU stay, and mortality. The use AG 100 system and AG tube in critically ill intubated patients is safe and effective in Pcuff control and SS drainage. Its protective role against VAP needs to be confirmed in a larger randomized trial. ClinicalTrials.gov NCT01550978. Date of registration: February 21, 2012.


Individually-addressable flip-chip AlInGaN micropixelated light emitting diode arrays with high continuous and nanosecond output power.
Micropixelated blue (470 nm) and ultraviolet (370 nm) AlInGaN light emitting diode ('micro-LED') arrays have been fabricated in flip-chip format with different pixel diameters (72 microm and 30 microm at, respectively, 100 and 278 pixels/mm(2)). Each micro-LED pixel can be individually-addressed and the devices possess a specially designed n-common contact incorporated to ensure uniform current injection and consequently uniform light emission across the array. The flip-chip micro-LEDs show, per pixel, high continuous output intensity of up to 0.55 microW/microm(2) (55 W/cm(2)) at an injection current density of 10 kA/cm(2) and can sustain continuous injection current densities of up to 12 kA/cm(2) before breakdown. We also demonstrate that nanosecond pulsed output operation of these devices with per pixel onaxis average peak intensity up to 2.9 microW/microm(2) (corresponding to energy of 45pJ per 22ns optical pulse) can be achieved. We investigate the pertinent performance characteristics of these arrays for micro-projection applications, including the prospect of integrated optical pumping of organic semiconductor lasers.


STEM study of Li4 Ti5 O12 anode material modified with Ag nanoparticles.
Comprehensive scanning transmission electron microscopy (STEM) analysis of Li4 Ti5 O12 (LTO) powder modified by deposited Ag nanoparticles was performed. Nanocomposite powders with Ag content of 1 wt.%, 4 wt.%, 10 wt.% were fabricated in a chemical process from suspensions of Ag and LTO. Apart from the STEM results, the presence of pure silver on the surface of the ceramic powder was confirmed by XRD and XPS analyses. The silver particles deposited on the LTO particles were characterized using the EDS mapping technique. The quantified results of the EDS mapping showed a relatively homogenous distribution of silver nanoparticles on the powder surface for every metal content. The mean diameter of the nanoparticles deposited on the LTO powder was about 4 nm in all cases. An increase in the Ag content during chemical surface modification did not cause changes in the microstructure. Focusing on an analysis of the metallic nanoparticles on the ceramic powder, electron tomography was used as an investigative technique. A very precise analysis of three-dimensional nanostructures is desirable for a comprehensive analysis of complex materials. The quantified analysis of the Ag nanoparticles visualized using electron tomography confirmed the results of the size measurements taken from the two-dimensional EDS maps.