EGFR point mutation in non-small cell lung cancer is occasionally accompanied by a second mutation or amplification.
Activating mutations of EGFR are found frequently in a subgroup of patients with non-small cell lung cancer (NSCLC) and are highly correlated with the response to gefitinib and erlotinib. In the present study, we searched for mutations of EGFR, HER2 and KRAS in 264 resected primary NSCLC from Japanese patients and determined whether there is a correlation between genetic alterations of these genes and clinicopathological factors, together with 85 tumors that we reported previously. EGFR mutations were found in 102 of the total 349 tumors, and seven tumors had two missense mutations. Reverse transcription-polymerase chain reaction of EGFR and subsequent subcloning analyses identified that the double mutations occurred in the same allele. Furthermore, in 202 NSCLC analyzed by Southern blotting, we identified 11 tumors with gene amplification of EGFR, with eight tumors containing a mutation in EGFR. Sequence analysis detected only weak or no signals of the wild-type allele in the eight tumors, strongly suggesting that the mutated allele was amplified selectively. These findings indicate that a dual genetic change of EGFR can occur in the same allele either with a possible second-hit mutation or with amplification, which may imply a more selective growth advantage in a cancer cell. Meanwhile, HER2 mutations and amplifications were found in six of 349 tumors and three of 202 tumors, respectively, and KRAS mutations in 21 of 349 tumors. Mutations of the EGFR and HER2 genes were more frequently found in female never or light-smoking patients with adenocarcinoma, and there were no tumors that had two or more mutations simultaneously among EGFR, HER2 and KRAS. The current study further demonstrates that a double genetic event in EGFR can occasionally occur in lung cancer, thus providing new clues for understanding the involvement of epidermal growth factor receptor signaling cascades in the pathogenesis of NSCLC.


Generation of two induced pluripotent stem cell lines (XACHi0010-A, XACHi0011-A) from a Chinese family with combined oxidative phosphorylation deficiency carrying homozygous and heterozygous C1QBP-L275F mutation.
Induced pluripotent stem cell lines (iPSCs) were generated from peripheral blood mononuclear cells (PBMCs) isolated from the peripheral blood of a 14 year-old boy and his mother using same protocols. Diagnosis of combined oxidative phosphorylation deficiency (COXPD) was established after identifying a homozygous c.823C > T(p.L275F) variant in C1QBP gene carried by the boy, inherited from his asymptomatic consanguineous parents carrying this heterozygous variant. PBMCs were reprogrammed using non-integrative sendai viral vectors containing reprogramming factors OCT4, SOX2, KLF4 and C-MYC. iPSCs were shown to express pluripotent markers, have trilineage differentiation potential, carry C1QBP-L275F mutation, have a normal karyotype. These lines are useful tools for studying the pathophysiological mechanism of COXPD.


Androgen-dependent regulation of human MUC1 mucin expression.
MUC1 mucin is transcriptionally regulated by estrogen, progesterone, and glucocorticoids. Our objective was to determine whether androgen receptor (AR) activation regulates expression of MUC1. The following breast and prostatic cell lines were phenotyped and grouped according to AR and MUC1 protein expression: 1) AR+MUC1 + [DAR17+19 (AR transfectants of DU-145), ZR-75-1, MDA-MB-453, and T47D]; 2) AR-MUC1 + [DZeo1 (AR-vector control), DU-145, BT20, MDA-MB-231, and MCF7]; 3) AR+MUC1 - (LNCaP and LNCaP-r). Cell proliferation was determined using the MTT assay in the presence of synthetic androgen R1881, 0.1 microM to 1 microM. Cell surface MUC1 expression was determined by flow cytometry in the presence or absence of oestradiol, medroxy progesterone acetate or R1881, with and without 4 hydroxy-flutamide (4-OH), a nonsteroidal AR antagonist. The functional significance of MUC1 expression was investigated with a cell-cell aggregation assay. Only AR+MUC1 + cell lines showed a significant increase in MUC1 expression with AR activation (P (range) =.01 to .0001), reversed in the presence of 4-OHF. Cell proliferation was unaffected. Increased expression of MUC1 was associated with a significant (P (range) = .002 to .001) reduction in cell-cell adhesion. To our knowledge, this is the first description of androgen-dependent regulation of MUC1 mucin. This is also functionally associated with decreased cell-cell adhesion, a recognised feature of progressive malignancy. These findings have important implications for physiological and pathological processes.


Genome-wide characterization of gene expression variations and DNA copy number changes in prostate cancer cell lines.
The aim of this study was to characterize gene expression and DNA copy number profiles in androgen sensitive (AS) and androgen insensitive (AI) prostate cancer cell lines on a genome-wide scale. Gene expression profiles and DNA copy number changes were examined using DNA microarrays in eight commonly used prostate cancer cell lines. Chromosomal regions with DNA copy number changes were identified using cluster along chromosome (CLAC). There were discrete differences in gene expression patterns between AS and AI cells that were not limited to androgen-responsive genes. AI cells displayed more DNA copy number changes, especially amplifications, than AS cells. The gene expression profiles of cell lines showed limited similarities to prostate tumors harvested at surgery. AS and AI cell lines are different in their transcriptional programs and degree of DNA copy number alterations. This dataset provides a context for the use of prostate cancer cell lines as models for clinical cancers.


Chromatin-associated degradation is defined by UBXN-3/FAF1 to safeguard DNA replication fork progression.
The coordinated activity of DNA replication factors is a highly dynamic process that involves ubiquitin-dependent regulation. In this context, the ubiquitin-directed ATPase CDC-48/p97 recently emerged as a key regulator of chromatin-associated degradation in several of the DNA metabolic pathways that assure genome integrity. However, the spatiotemporal control of distinct CDC-48/p97 substrates in the chromatin environment remained unclear. Here, we report that progression of the DNA replication fork is coordinated by UBXN-3/FAF1. UBXN-3/FAF1 binds to the licensing factor CDT-1 and additional ubiquitylated proteins, thus promoting CDC-48/p97-dependent turnover and disassembly of DNA replication factor complexes. Consequently, inactivation of UBXN-3/FAF1 stabilizes CDT-1 and CDC-45/GINS on chromatin, causing severe defects in replication fork dynamics accompanied by pronounced replication stress and eventually resulting in genome instability. Our work identifies a critical substrate selection module of CDC-48/p97 required for chromatin-associated protein degradation in both Caenorhabditis elegans and humans, which is relevant to oncogenesis and aging.


Grid computing and biomolecular simulation.
Biomolecular computer simulations are now widely used not only in an academic setting to understand the fundamental role of molecular dynamics on biological function, but also in the industrial context to assist in drug design. In this paper, two applications of Grid computing to this area will be outlined. The first, involving the coupling of distributed computing resources to dedicated Beowulf clusters, is targeted at simulating protein conformational change using the Replica Exchange methodology. In the second, the rationale and design of a database of biomolecular simulation trajectories is described. Both applications illustrate the increasingly important role modern computational methods are playing in the life sciences.


Optical and thermal responses of silicene in Xene heterostructures.
Stabilization of silicene and preservation of its structural and electronic properties are essential for its processing and future integration into devices. The stacking of silicene on stanene, creating a Xene-based heterostructure, proves to be a viable new route in this respect. Here we demonstrate the effectiveness of a stanene layer in breaking the strong interaction between silicene and the Ag(111) substrate. The role of stanene as a 'buffer' layer is investigated by analyzing the optical response of epitaxial silicene through both power-dependent Raman spectroscopy and reflectivity measurements in the near infrared (NIR)-ultraviolet (UV) spectral range. Finally, we point out a Xene-induced shift of the silver plasma edge that paves the way for the development of a new approach to engineering the metal plasmonic response.


Prognostic Significance of Tumor-Infiltrating Lymphocytes in Patients With Pancreatic Ductal Adenocarcinoma Treated With Neoadjuvant Chemotherapy.
The aim of this study was to examine tumor-infiltrating lymphocytes (TILs) and their prognostic value in patients with pancreatic ductal adenocarcinoma (PDAC) after neoadjuvant therapy. Intratumoral CD4, CD8, and FOXP3 lymphocytes were examined by immunohistochemistry using a computer-assisted quantitative analysis in 136 PDAC patients who received neoadjuvant therapy and pancreaticoduodenectomy. The results were correlated with clinicopathological parameters and survival. High CD4 TILs in treated PDAC were associated with high CD8 TILs (P = 0.003), differentiation (P = 0.04), and a lower frequency of recurrence (P = 0.02). Patients with high CD4 TILs had longer disease-free survival and overall survival (OS) than did patients with low CD4 TILs (P < 0.01). The median OS of patients with a high CD8/FOXP3 lymphocyte ratio (39.5 [standard deviation, 6.1] months) was longer than that of patients with a low CD8/FOXP3 lymphocyte ratio (28.3 [standard deviation, 2.3] months; P = 0.01). In multivariate analysis, high CD4 TILs were an independent prognostic factor for disease-free survival (hazard ratio, 0.49; 95% confidence interval, 0.30-0.81; P = 0.005) and OS (hazard ratio, 0.54; 95% confidence interval, 0.33-0.89; P = 0.02). High level of CD4 lymphocytes is associated with tumor differentiation and lower recurrence and is an independent prognostic factor for survival in PDAC patients treated with neoadjuvant therapy.


In Vivo Morphology of the Optic Nerve and Retina in Patients With Parkinson's Disease.
To investigate optic nerve (ON) and macular morphology in patients with Parkinson's disease (PD) using spectral-domain optical coherence tomography (SD-OCT). Twenty-five participants with PD (19 males and 6 females; mean age 60.79; SD ± 9.24) and 25 sex-, age-, ethnicity-, and refraction-matched healthy controls. A high-resolution SD-OCT device was used to acquire scans in 25 participants with PD (mean age 60.79; ± SD 9.24) and 25 sex-, age-, ethnicity-, and refraction-matched healthy controls. Main outcome measures included optic nerve head parameters (disc/cup diameters/areas, cup/rim volumes, cup depth, cup/disc ratio; peripapillary retinal nerve fiber layer [ppRNFL] thickness), retinal thickness (in inner and outer annuli around the foveal center) and thickness of individual retinal layers. Our study showed significant ppRNFL thinning in PD patients in all quadrants (P < 0.05) associated with a shallower optic cup (P = 0.03) as compared with controls. Foveal remodelling with retinal thinning (nasal and temporal segments in both annuli; and superior segment in outer annulus; P < 0.05), foveal pit widening (P = 0.05), central outer plexiform layer (OPL) thickening (P < 0.001), and nasal RPE thinning (P < 0.001) was also found in PD. The differences were more obvious in hemiretinae related to the predominantly affected cerebral hemisphere. Changes were more pronounced in advanced stages and longer PD duration. Optic nerve changes in PD are likely to be caused by primary neurodegeneration. Central retinal thinning, pit widening, central OPL thickening, and RPE thinning indicate foveal remodelling. Specific changes of the fovea and thinning of individual retinal layers, correlating with disease severity and duration, indicate that ON and retinal changes have potential to be used as biomarkers for PD.


Aggregatibacter actinomycetemcomitans leukotoxin: a powerful tool with capacity to cause imbalance in the host inflammatory response.
Aggregatibacter actinomycetemcomitans has been described as a member of the indigenous oral microbiota of humans, and is involved in the pathology of periodontitis and various non-oral infections. This bacterium selectively kills human leukocytes through expression of leukotoxin, a large pore-forming protein that belongs to the Repeat in Toxin (RTX) family. The specificity of the toxin is related to its prerequisite for a specific target cell receptor, LFA-1, which is solely expressed on leukocytes. The leukotoxin causes death of different leukocyte populations in a variety of ways. It activates a rapid release of lysosomal enzymes and MMPs from neutrophils and causes apoptosis in lymphocytes. In the monocytes/macrophages, the toxin activates caspase-1, a cysteine proteinase, which causes a proinflammatory response by the activation and secretion of IL-1β and IL-18. A specific clone (JP2) of A. actinomycetemcomitans with enhanced leukotoxin expression significantly correlates to disease onset in infected individuals. Taken together, the mechanisms by which this toxin kills leukocytes are closely related to the pathogenic mechanisms of inflammatory disorders, such as periodontitis. Therapeutic strategies targeting the cellular and molecular inflammatory host response in periodontal diseases might be a future treatment alternative.


Effect of Bisphenol A Glycol Methacrylate on Virulent Properties of Streptococcus mutans UA159.
Bisphenol A glycidyl methacrylate (bis-GMA), which is released into the oral environment by dental composites through incomplete polymerization, hydrolysis, and mechanical degradation, can significantly influence oral ecology around resin-based materials. The purpose of this study was to investigate how bis-GMA changes the virulence properties of Streptococcus mutans, a major cariogenic bacterium in humans. The results show that bis-GMA not only inhibited the planktonic growth of cells in medium containing glucose, fructose, or mannose, but also reduced the viability of S. mutans. However, the presence of bis-GMA increased sugar transport and intracellular polysaccharide accumulation in S. mutans, thereby increasing the potential of cell persistence. In addition, bis-GMA could enhance S. mutans's adhesion to hard surfaces and glucan synthesis, which could contribute to biofilm formation. Although free bis-GMA made cells vulnerable to acidic stress, it also provided increased resistance to hydrogen peroxide, which might confer an advantage in competition with other oral microorganisms during the early stage of biofilm development. Interestingly, the presence of bis-GMA did not change the ability of S. mutans to interact with saliva. The results suggest that leachable bis-GMA could contribute to biofilm-related secondary dental caries at the marginal interface between resin-based materials and teeth by altering the virulent properties of S. mutans, although bis-GMA reduced the planktonic growth and viability of S. mutans.


Integrons.
Integrons are genetic elements able to acquire and rearrange open reading frames (ORFs) embedded in gene cassette units and convert them to functional genes by ensuring their correct expression. They were originally identified as a mechanism used by Gram-negative bacteria to collect antibiotic resistance genes and express multiple resistance phenotypes in synergy with transposons. More recently, their role has been broadened with the discovery of chromosomal integron (CI) structures in the genomes of hundreds of bacterial species. This review focuses on the resources carried in these elements, on their unique recombination mechanisms, and on the different mechanisms controlling the cassette dynamics. We discuss the role of the toxin/antitoxin (TA) cassettes for the stabilization of the large cassette arrays carried in the larger CIs, known as superintegrons. Finally, we explore the central role played by single-stranded DNA in the integron cassette dynamics in light of the recent discovery that the integron integrase expression is controlled by the SOS response.


Flipping the world upside down: Using eye tracking in virtual reality to study visual search in inverted scenes.
Image inversion is a powerful tool for investigating cognitive mechanisms of visual perception. However, studies have mainly used inversion in paradigms presented on twodimensional computer screens. It remains open whether disruptive effects of inversion also hold true in more naturalistic scenarios. In our study, we used scene inversion in virtual reality in combination with eye tracking to investigate the mechanisms of repeated visual search through three-dimensional immersive indoor scenes. Scene inversion affected all gaze and head measures except fixation durations and saccade amplitudes. Our behavioral results, surprisingly, did not entirely follow as hypothesized: While search efficiency dropped significantly in inverted scenes, participants did not utilize more memory as measured by search time slopes. This indicates that despite the disruption, participants did not try to compensate the increased difficulty by using more memory. Our study highlights the importance of investigating classical experimental paradigms in more naturalistic scenarios to advance research on daily human behavior.


Supplementation of sows with L-Arginine during gestating and lactation affects muscle traits of offspring related with postnatal growth and meat quality: From conception to consumption.
This study investigated the effect of dietary inclusion of 25 g/day of L-Arginine (n = 7) or iso‑nitrogenous amounts of alanine (n = 6) from d 30 of gestation to d 28 of lactation of sows on performance, muscle traits and meat quality in offspring. From each litter, heaviest and smallest littermate of both sexes were reared from d 28 and slaughtered at d 140 in accordance with a 23factorial design. A response to L-Arginine were obtained on small females where L-Arginine increased birth weight, however this effect disappeared at weaning. L-Arginine increased daily gain by 7% and increased the cross-sectional area of the M. semitendinosus in small females by 14%, suggesting an increased lean ratio. Mechanistic studies showed firstly, that small female littermates had increased number of muscle fibres (myogenesis) after L-Arginine treatment (11%) and secondly increased total DNA (12%) as a consequence of satellite cell proliferation. Traits describing tenderness seem to be affected by L-Arginine but further studies are needed.


Nail gun injuries among construction workers.
Pneumatic nail guns greatly increase worker productivity and are extensively used in wood frame building construction, with especially high use in residential construction. One surveillance report of nail gun injuries in Washington State has been published; however, other literature consists largely of case reports and case series in trauma journals. The major objective of the current study was to investigate the occurrence of nail gun-associated injuries among construction workers and to identify preventable work-related factors associated with these injuries. Nail gun-related injuries occurring among a cohort of 13,347 carpenters in Ohio who worked union hours during the time period January 1, 1994, until September 30, 1997, were identified by matching the cohort with workers' compensation claims made to the Ohio Bureau of Workers' Compensation. We also analyzed workers' compensation claims for North Carolina Home Builders Association members for the period July 1996-November 1999 to identify nail gun-related injuries. Analyses included stratified analyses of claims by nature and body part injured, calculation of nail gun injury rates, and analyses of free text descriptions of injuries. Overall, nail gun injuries were responsible for 3.9 percent of workers' compensation claims with 8.3 percent to 25.5 percent of claims involving paid lost work time. The overall rate of nail gun injuries (cases per 200,000 work hours) was 0.33 in North Carolina and 0.26 in Ohio, reflecting the greater concentration of wood frame construction workers in the North Carolina population studied. Higher rates of injury were observed for carpenters in North Carolina and among residential carpenters in Ohio. The predominant body part injured was the hands/fingers, with 80 to 89 percent of injuries being nail punctures. Analyses of free text information for puncture injuries found approximately 70 percent of injuries to occur during the framing/sheathing stage of construction. Our data suggest that approximately 69 percent of puncture injuries may be due to an inadvertent gun discharge or misfire, preventable in large part by the use of sequential triggers. Worker training and education also are important components of nail gun injury prevention.


Contextual modulation involves suppression and facilitation from the center and the surround.
In psychophysics, cross-orientation suppression (XOS) and cross-orientation facilitation (XOF) have been measured by investigating mask configuration on the detection threshold of a centrally placed patch of sine-wave grating. Much of the evidence for XOS and XOF comes from studies using low and high spatial frequencies, respectively, where the interactions are thought to arise from within (XOS) and outside (XOF) the footprint of the classical receptive field. We address the relation between these processes here by measuring the effects of various sizes of superimposed and annular cross-oriented masks on detection thresholds at two spatial scales (1 and 7 c/deg) and on contrast increment thresholds at 7 c/deg. A functional model of our results indicates the following (1) XOS and XOF both occur for superimposed and annular masks. (2) XOS declines with spatial frequency but XOF does not. (3) The spatial extent of the interactions does not scale with spatial frequency, meaning that surround-effects are seen primarily at high spatial frequencies. (4) There are two distinct processes involved in XOS: direct divisive suppression and modulation of self-suppression. (5) Whether XOS or XOF wins out depends upon their relative weights and mask contrast. These results prompt enquiry into the effect of spatial frequency at the single-cell level and place new constraints on image-processing models of early visual processing.


Adaptive immunity in cancer immunology and therapeutics.
The vast genetic alterations characteristic of tumours produce a number of tumour antigens that enable the immune system to differentiate tumour cells from normal cells. Counter to this, tumour cells have developed mechanisms by which to evade host immunity in their constant quest for growth and survival. Tumour-associated antigens (TAAs) are one of the fundamental triggers of the immune response. They are important because they activate, via major histocompatibility complex (MHC), the T cell response, an important line of defense against tumourigenesis. However, the persistence of tumours despite host immunity implies that tumour cells develop immune avoidance. An example of this is the up-regulation of inhibitory immune checkpoint proteins, by tumours, which induces a form of self-tolerance. The majority of monoclonal antibodies in clinical practice have been developed to target tumour-specific antigens. More recently there has been research in the down-regulation of immune checkpoint proteins as a way of increasing anti-tumour immunity.


The role of ridges in the formation and longevity of flat slabs.
Flat-slab subduction occurs when the descending plate becomes horizontal at some depth before resuming its descent into the mantle. It is often proposed as a mechanism for the uplifting of deep crustal rocks ('thick-skinned' deformation) far from plate boundaries, and for causing unusual patterns of volcanism, as far back as the Proterozoic eon. For example, the formation of the expansive Rocky Mountains and the subsequent voluminous volcanism across much of the western USA has been attributed to a broad region of flat-slab subduction beneath North America that occurred during the Laramide orogeny (80-55 million years ago). Here we study the largest modern flat slab, located in Peru, to better understand the processes controlling the formation and extent of flat slabs. We present new data that indicate that the subducting Nazca Ridge is necessary for the development and continued support of the horizontal plate at a depth of about 90 kilometres. By combining constraints from Rayleigh wave phase velocities with improved earthquake locations, we find that the flat slab is shallowest along the ridge, while to the northwest of the ridge, the slab is sagging, tearing, and re-initiating normal subduction. On the basis of our observations, we propose a conceptual model for the temporal evolution of the Peruvian flat slab in which the flat slab forms because of the combined effects of trench retreat along the Peruvian plate boundary, suction, and ridge subduction. We find that while the ridge is necessary but not sufficient for the formation of the flat slab, its removal is sufficient for the flat slab to fail. This provides new constraints on our understanding of the processes controlling the beginning and end of the Laramide orogeny and other putative episodes of flat-slab subduction.


Continuous and intermittent CSF diversion after subarachnoid hemorrhage: a pilot study.
We examine two accepted methods of managing cerebrospinal fluid (CSF) drainage in patients following subarachnoid hemorrhage (SAH). The first is intermittent CSF drainage when intracranial pressure (ICP) reaches a pre-defined threshold (monitor-first) and the second is continuous CSF drainage (drain-first) at set pressure thresholds. This pilot study is designed to determine if there is a cause for a randomized study of comparing the two methods. This prospective observational pilot study enrolled 37 patients with SAH and external ventricular drainage between October 2008 and August 2009. Patients were treated with one of two methods of ICP management (drain-first vs. monitor-first) according to the discretion of the admitting physician. There were no significant differences in baseline characteristics including age, gender, severity of neurological dysfunction, and radiographic findings between the two groups. The incidence of vasospasm was not different between the drain-first group (66.7%; 16 of 24 patients) and the monitor-first group (53.9%; 7 of 13 patients). This pilot study was neither powered, nor expected to detect a difference between groups. The results of this study provide support for the design and conduct of a randomized study to assess the impact of two methods of CSF diversion for patients with SAH.


Cultivation of lacrimal gland acinar cells in a microgravity environment.
A rotary cell-culture system (RCCS) allows the creation of a microgravity environment of low shear force, high-mass transfer and three-dimensional cell culture of various cell types. The aim of the study was to evaluate the growth pattern and the secretory function of rabbit lacrimal gland acinar cells in a microgravity environment using an RCCS. Lacrimal gland acinar cells from male New Zealand White rabbits were isolated and cultured in an RCCS up to 28 days. Cells were analysed by light and electron microscopy, and apoptosis was assessed by the TUNEL assay at days 7, 14, 21 and 28. Secretory function was tested by measuring the beta-hexosaminidase activity. After 7 days of culture, spheroidal aggregates were found inside the RCCS. The spheroids consisted of acinus-like cell conglomerates. Apoptotic centres inside the spheroids were observed at all time points by means of the TUNEL assay. Evaluation of the secretory function revealed beta-hexosaminidase release after carbachol stimulation which decreased over the culture period. A simulated microgravity environment promotes the development of three-dimensional cell spheroids containing viable acinar cells up to 28 days. Due to the evolving central apoptosis, it is unlikely that such simple three-dimensional cell communities can serve as tissue equivalents for clinical transplantation, but they promise opportunities for further applications in basic and applied cell research on lacrimal gland cells.


Single Mothers and Poverty in Japan: The Role of Intergenerational Coresidence.
Single mothers' relatively high levels of poverty are well documented, but the role that intergenerational coresidence may play in mitigating this disadvantage is not well understood. In this paper, we use multiple rounds of a large national survey between 1986 and 2007 (n = 67,252) to evaluate the extent to which income sharing via intergenerational coresidence limits poverty among single mothers in Japan. Results indicate that conventional poverty rates based on single-mother households overstate the prevalence of poverty among single mothers by 12-20 percent as a result of excluding those who are coresiding with parents. We find no evidence that the prevalence of intergenerational coresidence has changed in a way that would offset the poverty-increasing effect of growth in single parenthood. Finally, we show not only that shared income is the most important factor in limiting poverty among single mothers living with parents, but also that many single mothers are coresiding with parents who are themselves economically disadvantaged. We discuss the implications of our findings for understanding relationships between rapid family change and poverty in countries like Japan, where public income support for single mothers is limited and where family support via intergenerational coresidence is both common and normative.


Rapid formin-mediated actin-filament elongation is essential for polarized plant cell growth.
Formins are present in all eukaryotes and are essential for the creation of actin-based structures responsible for diverse cellular processes. Because multicellular organisms contain large formin gene families, establishing the physiological functions of formin isoforms has been difficult. Using RNAi, we analyzed the function of all 9 formin genes within the moss Physcomitrella patens. We show that plants lacking class II formins (For2) are severely stunted and composed of spherical cells with disrupted actin organization. In contrast, silencing of all other formins results in normal elongated cell morphology and actin organization. Consistent with a role in polarized growth, For2 are apically localized in growing cells. We show that an N-terminal phosphatase tensin (PTEN)-like domain mediates apical localization. The PTEN-like domain is followed by a conserved formin homology (FH)1-FH2 domain, known to promote actin polymerization. To determine whether apical localization of any FH1-FH2 domain mediates polarized growth, we performed domain swapping. We found that only the class II FH1-FH2, in combination with the PTEN-like domain, rescues polarized growth, because it cannot be replaced with a similar domain from a For1. We used in vitro polymerization assays to dissect the functional differences between these FH1-FH2 domains. We found that both the FH1 and the FH2 domains from For2 are required to mediate exceptionally rapid rates of actin filament elongation, much faster than any other known formin. Thus, our data demonstrate that rapid rates of actin elongation are critical for driving the formation of apical filamentous actin necessary for polarized growth.


Spatial analysis spotlighting early childhood leprosy transmission in a hyperendemic municipality of the Brazilian Amazon region.
More than 200,000 new cases of leprosy were reported by 105 countries in 2011. The disease is a public health problem in Brazil, particularly within high-burden pockets in the Amazon region where leprosy is hyperendemic among children. We applied geographic information systems and spatial analysis to determine the spatio-temporal pattern of leprosy cases in a hyperendemic municipality of the Brazilian Amazon region (Castanhal). Moreover, we performed active surveillance to collect clinical, epidemiological and serological data of the household contacts of people affected by leprosy and school children in the general population. The occurrence of subclinical infection and overt disease among the evaluated individuals was correlated with the spatio-temporal pattern of leprosy. The pattern of leprosy cases showed significant spatio-temporal heterogeneity (p<0.01). Considering 499 mapped cases, we found spatial clusters of high and low detection rates and spatial autocorrelation of individual cases at fine spatio-temporal scales. The relative risk of contracting leprosy in one specific cluster with a high detection rate is almost four times the risk in the areas of low detection rate (RR = 3.86; 95% CI = 2.26-6.59; p<0.0001). Eight new cases were detected among 302 evaluated household contacts: two living in areas of clusters of high detection rate and six in hyperendemic census tracts. Of 188 examined students, 134 (71.3%) lived in hyperendemic areas, 120 (63.8%) were dwelling less than 100 meters of at least one reported leprosy case, 125 (66.5%) showed immunological evidence (positive anti-PGL-I IgM titer) of subclinical infection, and 9 (4.8%) were diagnosed with leprosy (8 within 200 meters of a case living in the same area). Spatial analysis provided a better understanding of the high rate of early childhood leprosy transmission in this region. These findings can be applied to guide leprosy control programs to target intervention to high risk areas.


Early malaria resurgence in pre-elimination areas in Kokap Subdistrict, Kulon Progo, Indonesia.
Indonesia is among those countries committed to malaria eradication, with a continuously decreasing incidence of malaria. However, at district level the situation is different. This study presents a case of malaria resurgence Kokap Subdistrict of the Kulon Progo District in Yogyakarta Province, Java after five years of low endemicity. This study also aims to describe the community perceptions and health services delivery situation that contribute to this case. All malaria cases (2007-2011) in Kulon Progo District were stratified to annual parasite incidence (API). Two-hundred and twenty-six cases during an outbreak (May 2011 to April 2012) were geocoded by household addresses using a geographic information system (GIS) technique and clusters were identified by SaTScan software analysis (Arc GIS 10.1). Purposive random sampling was conducted on respondents living inside the clusters to identify community perceptions and behaviour related to malaria. Interviews were conducted with malaria health officers to understand the challenges of malaria surveillance and control. After experiencing three consecutive years with API less than 1 per thousand, malaria in Kokap subdistrict increased almost ten times higher than API in the district level and five times higher than national API. Malaria cases were found in all five villages in 2012. One primary and two secondary malaria clusters in Hargotirto and Kalirejo villages were identified during the 2011-2012 outbreak. Most of the respondents were positively aware with malaria signs and activities of health workers to prevent malaria, although some social economic activities could not be hindered. Return transmigrants or migrant workers entering to their villages, reduced numbers of village malaria workers and a surge in malaria cases in the neighbouring district contributed to the resurgence. Community perception, awareness and participation could constitute a solid foundation for malaria elimination in Kokap. However, decreasing number of village malaria workers and ineffective communication between primary health centres (PHCs) within boundary areas with similar malaria problems needs attention. Decentralization policy was allegedly the reason for the less integrated malaria control between districts, especially in the cross border areas. Malaria resurgence needs attention particularly when it occurs in an area that is entering the elimination phase.


Maternal and health care workers' perceptions of the effects of exclusive breastfeeding by HIV positive mothers on maternal and infant health in Blantyre, Malawi.
HIV-positive mothers are likely to exclusively breastfeed if they perceive exclusive breastfeeding (EBF) beneficial to them and their infants. Nevertheless, very little is known in Malawi about HIV-positive mothers' perceptions regarding EBF. In order to effectively promote EBF among these mothers, it is important to first understand their perceptions on benefits of exclusive breastfeeding. This study therefore, explored maternal and health care workers' perceptions of the effects of exclusive breastfeeding on HIV-positive mothers' health and that of their infants. This was a qualitative study within a larger project. Face-to-face in-depth interviews and focus group discussions using a semi- structured interview and focus group guide were conducted. Sixteen HIV-positive breastfeeding mothers, between 18 and 35 years old, were interviewed and data saturation was achieved. Two focus group discussions (FGDs) comprising of five and six adult women of unknown HIV status who were personal assistants to maternity patients, and one FGD with five nurse-midwives working in the maternity wards of Queen Elizabeth Central Hospital in Blantyre, Malawi, were also conducted. Thematic content data analysis was utilized. The study revealed more positive than negative perceived effects of exclusive breastfeeding. However, the fear of transmitting HIV to infants through breast milk featured strongly in the study participants' reports including those of the nurse-midwives. Only one nurse-midwife and a few HIV-positive mothers believed that EBF prevents mother-to-child transmission of HIV. Furthermore, participants, especially the HIV-positive mothers felt that exclusive breastfeeding leads to maternal ill- health and would accelerate their progression to full blown AIDS. While most participants considered exclusive breastfeeding as an important component of the wellbeing of their infants' health, they did not share the worldwide acknowledged benefits of exclusive breastfeeding in the prevention of mother-to-child transmission (PMTCT) of HIV. These results suggest a need for more breastfeeding education for all mothers, communities and nurse-midwives involved in breastfeeding counseling in the context of HIV infection. Maternal wellbeing promotion activities such as nutrition supplementation need to be included in all PMTCT of HIV programs.


Comparison of hypopnea definitions in lean patients with known obstructive sleep apnea hypopnea syndrome (OSAHS).
In the interest of improving inter-rater reliability and standardization between sleep laboratories, hypopnea definitions were recently changed to place less emphasis on arousal scoring and more emphasis on oxygen desaturations. We sought to determine whether these changes would affect detection and treatment of OSAHS in lean patients-a group known to desaturate less-than-obese patients. Thirty-five lean subjects (15 male, 20 women, five post-menopausal) diagnosed OSAHS and a documented benefit from treatment had diagnostic polysomnograms (PSG) originally scored using the American Academy of Sleep Medicine (AASM) rule from 1999 (referred to as "Rule C"). These patients had appropriate clinical care based on those results. PSG records were then re-scored in a randomized and blinded fashion utilizing hypopnea Rule A and B of the 2007 AASM guidelines. Baseline mean (SD) apnea hypopnea indices (AHI) for rules A, B, and C were 6.4 (3.1), 20.6 (8.2), and 26.9 (7.3), respectively (p < 0.0001). Mean (SD) BMI was 24.4 (1.0). By design, all subjects were treatment responders. Eighty-six percent with CPAP, 83% with oral appliance, and 100% with surgical intervention reported resolution of their initial daytime or sleep complaint. Post-treatment AHIs for rules A, B, and C were 0.8 (0.9), 1.8 (1.2) and 2.3 (1.6; p < 0.001). In all three scoring conditions, the AHI was reduced significantly with treatment (p < 0.001). A repeated measures ANOVA of the difference between scoring methods indicated statistically significant differences between all three strategies at both pre- and post-treatment (p < 0.001). Sleepiness on the Epworth sleepiness scale decreased from a mean of 10.9 (2.3) to 5.7 (1.3) with treatment (p < 0.001). This change in subjective rating of sleepiness was more strongly correlated with rules B and C (r = 0.6) and more modestly correlated with Rule A scoring (r = 0.4). Response to treatment was more tightly correlated with arousal based scoring rules B and C in this group of lean subjects. The1999 hypopnea rule was used at baseline to detect this cohort of patients with OSAHS that ultimately benefitted from treatment. Rule B detected OSAHS and correlated well with response to treatment, but many more were categorized as mild (5 < AHI < 15) at baseline. Since 40% of the subjects had an AHI less than 5 with Rule A, lack of sensitivity should be considered before applying Rule A to the scoring of sleep studies in lean patients.


The significance of leukemic cell volume distribution.
The use of electronic pulse spectroscopy to determine the volume distribution of a white blood cell fraction isolated by sedimentation from anticoagulated whole blood permits a rapid quantitative measure of size and dispersion of white blood cells in suspension. In patients without apparent hematologic diseases the white cell volume distribution presents as a broad peak. In patients with a large percentage of circulatory malignant cells, a single narrow volume peak is observed in acute leukemias and a broad and sometimes bimodal curve is observed in chronic leukemias. The volume of the leukemic cells is observed to shift in vivo after the start of effective chemotherapy as was predicted on the basis of a tissue culture model. The prognostic significance of the white cell volume distribution in leukemia and the use of a shift in size of leukemic cells after the start of therapy as a means of monitoring therapy are discussed.


ddcMD: A fully GPU-accelerated molecular dynamics program for the Martini force field.
We have implemented the Martini force field within Lawrence Livermore National Laboratory's molecular dynamics program, ddcMD. The program is extended to a heterogeneous programming model so that it can exploit graphics processing unit (GPU) accelerators. In addition to the Martini force field being ported to the GPU, the entire integration step, including thermostat, barostat, and constraint solver, is ported as well, which speeds up the simulations to 278-fold using one GPU vs one central processing unit (CPU) core. A benchmark study is performed with several test cases, comparing ddcMD and GROMACS Martini simulations. The average performance of ddcMD for a protein-lipid simulation system of 136k particles achieves 1.04 µs/day on one NVIDIA V100 GPU and aggregates 6.19 µs/day on one Summit node with six GPUs. The GPU implementation in ddcMD offloads all computations to the GPU and only requires one CPU core per simulation to manage the inputs and outputs, freeing up remaining CPU resources on the compute node for alternative tasks often required in complex simulation campaigns. The ddcMD code has been made open source and is available on GitHub at https://github.com/LLNL/ddcMD.


Image rotation and translation measurement based on double phase-encoded joint transform correlator.
An image rotation and translation measurement technology based on a double phase-encoded joint transform correlator (DPEJTC) is proposed. The reference and the target images are Fourier transformed. Then the magnitude of the Fourier-transformed reference (MFR) and target (MFT) images are multiplied with a high-pass emphasis filter and transformed from Cartesian space into polar space. Rotation between the reference and the target image is obtained by measuring the emphasized MFR and MFT in polar coordinates by the DPEJTC. The target image is rotated by the rotation angle in the inverse orientation to get the rotation-correction target image. Finally, translation between the reference and the target image is obtained through measuring the reference and the rotation-correction target image by the DPEJTC. Results based on digital computation are given to verify our proposal. A possible optical setup is suggested.


Evolution of near- and far-field optical properties of Au bipyramids upon epitaxial deposition of Ag.
Bimetallic plasmonic nanostructures provide composition and spatial distribution of the individual components in the nanostructure in addition to overall size and morphology as degrees of freedom for tuning near- and far-field optical responses. AgAuAg nanorods (NRs) generated through epitaxial deposition of Ag on the tips of Au bipyramids (BPs) are an important bimetallic model system whose longitudinal dipolar plasmon mode first shows a spectral blue-shift upon initial deposition of Ag on the Au BP tips followed by a red-shift after additional deposition of Ag. Here, we quantify the relative contributions from morphological and compositional effects to the far-field spectral shift of the longitudinal and vertical dipolar plasmon modes during the initial deposition of Ag and compare the near-field in Ag and AgAuAg NRs with lengths between L = 130 nm-280 nm under whitelight illumination through electromagnetic simulations. Subsequently, we experimentally characterize the near-field around AgAuAg NRs with lengths between L = 88.1-749.0 nm at a constant excitation wavelength of 1064 nm on a silicon (Si) support through scattering type near-field scanning microscopy (sNSOM). We detect Fabry-Perot resonance-like higher order multipolar plasmon resonances whose order and near-field pattern depends on the length and composition of the NRs as well as the refractive index of the ambient medium. We find that under oblique illumination higher order multipolar modes with an even symmetry dominate on the high refractive index Si substrate due to strong electromagnetic interactions between the NR and the substrate.


Children with albinism in African regions: their rights to 'being' and 'doing'.
Albinism is an inherited condition with a relatively high prevalence in populations throughout sub-Saharan Africa. People with oculocutaneous albinism have little or no pigment in their hair, skin and eyes; thus they are visually impaired and extremely sensitive to the damaging effect of the sun on their skin. Aside from the health implications of oculocutaneous albinism, there are also significant sociocultural risks. The impacts of albinism are particularly serious in areas that associate albinism with legend and folklore, leading to stigmatisation and discrimination. In regions of Africa those with albinism may be assaulted and sometimes killed for their body parts for use in witchcraft-related rites or to make 'lucky' charms. There is a dearth of research on the psychosocial aspects of albinism and particularly on how albinism impacts on the everyday lives of people with albinism. There is a growing recognition and acceptance in Africa that people with albinism should be considered disabled. Thomas's social-relational model of disability proposes it is essential to understand both the socio-structural barriers and restrictions that exclude disabled people (barriers to doing); and the social processes and practices which can negatively affect their psycho-emotional wellbeing (barriers to being). In this article, we combine a social model of disability with discussion on human rights to address the lacuna surrounding the psychosocial and daily experiences of people with albinism. Through using this combined framework we conclude that the rights of people with albinism in some regions of Africa are not being enacted. Our debate highlights the need to develop a holistic concept of rights for children and young people with albinism which sees human rights as indivisible. We illuminate some of the specific ways in which the lives of children with albinism could be improved by addressing 'barriers to being' and 'barriers to doing', at the heart of which requires a shift in attitude and action to address discrimination.


Efficient Display of Aspergillus niger β-Glucosidase on Saccharomyces cerevisiae Cell Wall for Aroma Enhancement in Wine.
The aim of this study was to evaluate the potential application of cell-surface-displayed β-glucosidase (BGL) in wine aroma enhancement. Gene cassettes for the surface display of Aspergillus niger BGL were constructed using different promoters ( GPD and SED1) and glycosylphosphatidylinositol (GPI) anchoring regions (Sag1, Sed1, and Cwp2). The differences in surface-display cassettes, the tolerance of the displayed BGL to typical winemaking conditions, and the hydrolysis capacity for the liberation of grape aroma glycosides were analyzed. Results revealed that simultaneous utilization of GPD promoter and Sed1 anchoring domain achieved the highest BGL activity. The displayed BGL exhibited relatively high activity at pH 3.0 and at glucose concentration below 2.5% (w/v), compared to commercial enzyme (AR 2000), but exhibited no significant difference under varying ethanol concentrations. Furthermore, the surface-displayed BGL presented better ability to release free terpenols compared to AR 2000. Therefore, a surface-display system could provide a new viable solution for enhancing wine aroma.


Integrated intravascular ultrasound and photoacoustic imaging scan head.
The combination of intravascular ultrasound and intravascular photoacoustic imaging has been proposed for improving the diagnosis of arterial diseases. We describe a novel scan-head design for implementing such multimodality imaging. The proposed device has the potential to achieve a sufficiently small size for clinical intravascular applications. The design aims for efficient image data acquisition for facilitating real-time three-dimensional imaging and reducing the required laser pulse repetition frequency. The integrated scan head consists of a single-element, ring-shaped transducer for sideward ultrasound transmission, a multimode fiber with a cone-shaped mirror for optical illumination, and a single polymer microring with mechanical scanning. The phantom imaging and some experimental results are presented. A microring array can be realized in the future to achieve high-frame-rate intravascular multimodality imaging.


Persistence in breast cancer disparities between African Americans and whites in Wisconsin.
Breast cancer (BC) mortality is higher in African American women compared to white women despite having a lower incidence. The reasons for this remain unclear, despite decades of research. Reducing BC health disparities is a priority but has had limited success. To assess progress in eliminating breast cancer-related health disparities in Wisconsin by comparing trends in breast cancer outcomes in African American and white women from 1995 to 2006 and comparing results nationally. Age-adjusted breast cancer (BC) incidence and stage data from the Wisconsin Cancer Reporting System and age-adjusted mortality data from National Center of Health Statistics were used to evaluate trends in incidence and mortality from 1995 to 2006 for African Americans and whites. The relative disparity was evaluated by rate ratios. Trends in distribution of in situ vs malignant disease were examined. National trend data were obtained from the National Cancer Institute (NCI) Surveillance, Epidemiology and End Results (SEER) database. Age-adjusted incidence decreased 10% in Wisconsin compared to 7% nationally. Incidence of BC was lower in African American compared to white women. BC mortality in African American women declined in Wisconsin, but remained higher than white females. Age-adjusted mortality in Wisconsin declined approximately 23%, matching national trends. Non age-adjusted stage data trended toward a decrease in malignant, but increased in situ disease. Despite an overall reduction in BC mortality from 1995 to 2006, a persistent disparity in mortality remains for African American women, demonstrating no significant progress in reducing BC health disparities.


Vasoactive intestinal peptide regulates cholesterol side-chain cleavage cytochrome P-450 (P-450scc) gene expression in granulosa cells from immature rat ovaries.
Vasoactive intestinal peptide (VIP), a neuropeptide present in ovarian nerves, has been previously shown to induce synthesis of the side-chain cleavage cytochrome P-450 enzyme which catalyzes the conversion of cholesterol to pregnenolone (the rate-limiting step in progesterone synthesis). In the present study we demonstrate, by means of a bovine 3'-specific P-450scc cDNA probe, that this VIP effect is exerted at least partially at the level of gene expression in cultured granulosa cells that were isolated from estrogen-primed, immature rats. The size and level of the 2.0 kilobase P-450scc mRNA species was assessed by Northern blot analysis, while the translatability of this mRNA was assayed by immunoisolation of the 35S-labeled P-450scc precursor protein translated from total RNA of control and stimulated granulosa cells. FSH was much more effective than VIP at increasing P-450scc mRNA concentrations in cultured granulosa cells, whereas secretin treatment was ineffective. The results suggest that, like FSH, the stimulatory effect of the neuropeptide VIP on ovarian progesterone secretion involves regulation of P-450scc gene expression during functional maturation of the prepubertal ovary.


The contribution of school experience to students' resilience following a terror-related homicide.
Schools have a significant effect on students' development, and serve as important social agencies for interventions for students facing disasters. However, little is known about the effect of students' school experience itself on their resilience when facing extreme negative events. The present study focused on students who were exposed to terror-related homicide with the aim of investigating the contribution of school climate resources to their resilience. Since resilience is associated not only with fewer negative outcomes, but also with positive change, the contribution of schools was studied as both inhibiting post-traumatic stress symptoms (PTS) and enhancing post-traumatic growth (PTG). A mixed-methods research design was used. The participants included 117 (52% girls) high school students (mean age = 14.54; SD = 1.49). Twenty-five of them were interviewed in addition to responding to the research questionnaires. Different aspects of the school climate were found to be associated with students' PTS and PTG, yielding two overarched factors explaining the school's role as a protective resource: sheltering and supporting. The former is associated with fewer PTS and the latter with higher PTG. The use of different resources for different forms of resilience is discussed.


Adhesion at calcium oxalate crystal surfaces and the effect of urinary constituents.
Kidney stones, aggregates of microcrystals, most commonly contain calcium oxalate monohydrate (COM) as the primary constituent. The aggregation of COM microcrystals and their attachment to epithelial cells are thought to involve adhesion at COM crystal surfaces, mediated by anionic molecules or urinary macromolecules. Identification of the most important functional group-crystal face adhesive combinations is crucial to understanding the stability of COM aggregates and the strength of their attachments to epithelial cell surfaces under flow in the renal tubules of the kidney. Here, we describe direct measurements of adhesion forces, by atomic force microscopy, between various functional groups and select faces of COM crystals immersed in aqueous media. Tip-immobilized carboxylate and amidinium groups displayed the largest adhesion forces, and the adhesive strength of the COM crystal faces decreased in the order (100) > (121) > (010), demonstrating that adhesion is sensitive to the structure and composition of crystal faces. The influence of citrate and certain urinary proteins on adhesion was examined, and it was curious that osteopontin, a suspected regulator of stone formation, increased the adhesion force between a carboxylate tip and the (100) crystal face. This behavior was unique among the various combinations of additives and COM crystal faces examined here. Collectively, the force measurements demonstrate that adhesion of functional groups and binding of soluble additives, including urinary macromolecules, to COM crystal surfaces are highly specific in nature, suggesting a path toward a better understanding of kidney stone disease and the eventual design of therapeutic agents.


Natural demodulation of two-dimensional fringe patterns. I. General background of the spiral phase quadrature transform.
It is widely believed, in the areas of optics, image analysis, and visual perception, that the Hilbert transform does not extend naturally and isotropically beyond one dimension. In some areas of image analysis, this belief has restricted the application of the analytic signal concept to multiple dimensions. We show that, contrary to this view, there is a natural, isotropic, and elegant extension. We develop a novel two-dimensional transform in terms of two multiplicative operators: a spiral phase spectral (Fourier) operator and an orientational phase spatial operator. Combining the two operators results in a meaningful two-dimensional quadrature (or Hilbert) transform. The new transform is applied to the problem of closed fringe pattern demodulation in two dimensions, resulting in a direct solution. The new transform has connections with the Riesz transform of classical harmonic analysis. We consider these connections, as well as others such as the propagation of optical phase singularities and the reconstruction of geomagnetic fields.


'It's a cultural expectation...' The pressure on medical trainees to work independently in clinical practice.
Medical trainees demonstrate a reluctance to ask for help unless they believe it is absolutely necessary, a situation which could impact on the safety of patients. This study aimed to develop a theoretical exploration of the pressure on medical trainees to be independent and to generate theory-based approaches to the implications for patient safety of this pressure towards independent working. In Phase 1, 88 teaching team members from internal and emergency medicine were observed during clinical activities (216 hours), and 65 participants completed brief interviews. In Phase 2, 36 in-depth interviews were conducted using video vignettes. Data collection and analysis employed grounded theory methodology. Participants conceived that the pressure towards independence in clinical work originated in trainees' desire to lay claim to the identity of a doctor (as a member of a group of autonomous high achievers), and in organisational issues such as heavy workloads and constant evaluations. The identity and organisational issues related to the pressure towards independence were explored through the lenses of established theories from education and psychology. Consideration of Lave and Wenger's situated learning theory suggests that giving attention to the 'independent doctor' ideal, through measures such as involving trainees when their supervisors ask for help, could impact the safety of teaching team practice. Amalberti et al.'s migration model explains how pressures to maximise productivity and individual gain may cause teaching teams to migrate beyond the boundaries of safe practice and suggests that managing triggers (such as workload and high-stakes evaluations) for violations of safe practice might improve safety. Implementation and evaluation of these theory-based approaches to the safety of teaching team practice would contribute to a better understanding of the links between trainee independence and patient safety.


IoT-Based Remote Pain Monitoring System: From Device to Cloud Platform.
Facial expressions are among behavioral signs of pain that can be employed as an entry point to develop an automatic human pain assessment tool. Such a tool can be an alternative to the self-report method and particularly serve patients who are unable to self-report like patients in the intensive care unit and minors. In this paper, a wearable device with a biosensing facial mask is proposed to monitor pain intensity of a patient by utilizing facial surface electromyogram (sEMG). The wearable device works as a wireless sensor node and is integrated into an Internet of Things (IoT) system for remote pain monitoring. In the sensor node, up to eight channels of sEMG can be each sampled at 1000 Hz, to cover its full frequency range, and transmitted to the cloud server via the gateway in real time. In addition, both low energy consumption and wearing comfort are considered throughout the wearable device design for long-term monitoring. To remotely illustrate real-time pain data to caregivers, a mobile web application is developed for real-time streaming of high-volume sEMG data, digital signal processing, interpreting, and visualization. The cloud platform in the system acts as a bridge between the sensor node and web browser, managing wireless communication between the server and the web application. In summary, this study proposes a scalable IoT system for real-time biopotential monitoring and a wearable solution for automatic pain assessment via facial expressions.


Human tumors instigate granulin-expressing hematopoietic cells that promote malignancy by activating stromal fibroblasts in mice.
Systemic instigation is a process by which endocrine signals sent from certain tumors (instigators) stimulate BM cells (BMCs), which are mobilized into the circulation and subsequently foster the growth of otherwise indolent carcinoma cells (responders) residing at distant anatomical sites. The identity of the BMCs and their specific contribution or contributions to responder tumor growth have been elusive. Here, we have demonstrated that Sca1+ cKit- hematopoietic BMCs of mouse hosts bearing instigating tumors promote the growth of responding tumors that form with a myofibroblast-rich, desmoplastic stroma. Such stroma is almost always observed in malignant human adenocarcinomas and is an indicator of poor prognosis. We then identified granulin (GRN) as the most upregulated gene in instigating Sca1+ cKit- BMCs relative to counterpart control cells. The GRN+ BMCs that were recruited to the responding tumors induced resident tissue fibroblasts to express genes that promoted malignant tumor progression; indeed, treatment with recombinant GRN alone was sufficient to promote desmoplastic responding tumor growth. Further, analysis of tumor tissues from a cohort of breast cancer patients revealed that high GRN expression correlated with the most aggressive triple-negative, basal-like tumor subtype and reduced patient survival. Our data suggest that GRN and the unique hematopoietic BMCs that produce it might serve as novel therapeutic targets.


An unusual intraerythrocytic parasite of Parablennius cornutus from South Africa.
An intertidal horned blenny, Parablennius cornutus, captured at De Hoop Nature Reserve, South Africa, was found to harbor an unusual blood parasite and the haemogregarine Haemogregarina bigemina. In Giemsa-stained blood films, the enigmatic parasite occurred primarily as intraerythrocytic ringlike stages, with unstained centers and peripheral bands of beaded chromatin, not unlike Haemohormidium spp. Larger forms of the same organism stained pink with Giemsa, with nuclei occurring as 4-8 minute structures around the parasite body or distributed within it. These larger parasites apparently segmented into up to 8 individuals that were rounded or oval with deep-stained, comma-shaped or polar regions surrounding blue cytoplasm. Extracellular, binucleate, sporelike structures in clusters of as many as 16 individuals were also seen in blood films. Praniza larvae of the isopod Gnathia africana were seen in histological sections of gill tissue. Examination of spleen tissue by transmission electron microscopy showed intraerythrocytic organisms with ultrastructural characteristics like those of Haematractidium scombri, namely, a single boundary membrane, sometimes closely apposed nuclei with nucleoli, and profiles of dense material of variable structure. It is concluded that the parasite is probably related to Haemohormidium spp. and H. scombri, but it also shares features with some Microsporida.


Diffusive Gradients in Thin Films Can Predict the Toxicity of Metal Mixtures to Two Microalgae: Validation for Environmental Monitoring in Antarctic Marine Conditions.
Anthropogenic contamination in the Antarctic near-shore marine environment is a challenge for environmental managers because of its isolation, high costs associated with monitoring and remediation activities, and the current lack of Antarctic-specific ecotoxicological data. The present study investigated the application of diffusive gradients in thin films (DGT) with a Chelex-100 binding resin for metal contaminant assessment in Antarctic marine conditions. Diffusion coefficients for cadmium (Cd), copper (Cu), nickel (Ni), lead (Pb), and zinc (Zn), determined at 1 °C, ranged between 2.1 and 2.6×10-6 cm2 /s and were up to 32% lower than those derived by theoretical calculations. Competition of metals on the DGT binding resin was observed at subsaturation concentrations, reducing the effective capacity for metal uptake by approximately 60%. The lability of the dissolved (0.45 µm filterable) Cd, Cu, Pb, and Zn metal fraction to DGT was generally >90% and unaffected by the presence of the Antarctic marine microalga Phaeocystis antarctica. Both DGT and dissolved metal concentrations gave equivalent mixture toxicity predictions in independent action and concentration addition models to P. antarctica and Cryothecomonas armigera; that is, predictions using DGT-labile concentrations also showed antagonism to P. antarctica, which agrees with previously determined mixture interactivity. The benefits of DGT over traditional sampling techniques (i.e., discrete water sampling) include lower method detection limits (MDLs), in situ assessment, and time-averaged concentrations which capture pulses of contamination typical of the Antarctic near-shore marine environment. The present study provides MDLs and recommended minimum deployment times to guide field deployments in Antarctica. Environ Toxicol Chem 2019;38:1323-1333. © 2019 SETAC.


Humanization and expression of IgG and IgM antibodies in plants as potential diagnostic reagents for Valley Fever.
Monoclonal antibodies (mAbs) are important proteins used in many life science applications, from diagnostics to therapeutics. High demand for mAbs for different applications urges the development of rapid and reliable recombinant production platforms. Plants provide a quick and inexpensive system for producing recombinant mAbs. Moreover, when paired with an established platform for mAb discovery, plants can easily be tailored to produce mAbs of different isotypes against the same target. Here, we demonstrate that a hybridoma-generated mouse mAb against chitinase 1 (CTS1), an antigen from Coccidioides spp., can be biologically engineered for use with serologic diagnostic test kits for coccidioidomycosis (Valley Fever) using plant expression. The original mouse IgG was modified and recombinantly produced in glycoengineered Nicotiana benthamiana plants via transient expression as IgG and IgM isotypes with human kappa, gamma, and mu constant regions. The two mAb isotypes produced in plants were shown to maintain target antigen recognition to CTS1 using similar reagents as the Food and Drug Administration (FDA)-approved Valley Fever diagnostic kits. As none of the currently approved kits provide antibody dilution controls, humanization of antibodies that bind to CTS1, a major component of the diagnostic antigen preparation, may provide a solution to the lack of consistently reactive antibody controls for Valley Fever diagnosis. Furthermore, our work provides a foundation for reproducible and consistent production of recombinant mAbs engineered to have a specific isotype for use in diagnostic assays.


Noise in restaurants: levels and mathematical model.
Noise affects the dining atmosphere and is an occupational hazard to restaurant service employees worldwide. This paper examines the levels of noise in dining areas during peak hours in different types of restaurants in Hong Kong SAR, China. A mathematical model that describes the noise level in a restaurant is presented. The 1-h equivalent continuous noise level (L(eq,1-h)) was measured using a Type-1 precision integral sound level meter while the occupancy density, the floor area of the dining area, and the ceiling height of each of the surveyed restaurants were recorded. It was found that the measured noise levels using Leq,1-h ranged from 67.6 to 79.3 dBA in Chinese restaurants, from 69.1 to 79.1 dBA in fast food restaurants, and from 66.7 to 82.6 dBA in Western restaurants. Results of the analysis of variance show that there were no significant differences between means of the measured noise levels among different types of restaurants. A stepwise multiple regression analysis was employed to determine the relationships between geometrical and operational parameters and the measured noise levels. Results of the regression analysis show that the measured noise levels depended on the levels of occupancy density only. By reconciling the measured noise levels and the mathematical model, it was found that people in restaurants increased their voice levels when the occupancy density increased. Nevertheless, the maximum measured hourly noise level indicated that the noise exposure experienced by restaurant service employees was below the regulated daily noise exposure value level of 85 dBA.


Propagation of scrapie in peripheral nerves after footpad infection in normal and neurotoxin exposed hamsters.
As is known from various animal models, the spread of agents causing transmissible spongiform encephalopathies (TSE) after peripheral infection affects peripheral nerves before reaching the central nervous system (CNS) and leading to a fatal end of the disease. The lack of therapeutic approaches for TSE is partially due to the limited amount of information available on the involvement of host biological compartments and processes in the propagation of the infectious agent. The in vivo model presented here can provide information on the spread of the scrapie agent via the peripheral nerves of hamsters under normal and altered axonal conditions. Syrian hamsters were unilaterally footpad (f.p.) infected with scrapie. The results of the spatiotemporal ultrasensitive immunoblot-detection of scrapie-associated prion protein (PrP(Sc)) in serial nerve segments of both distal sciatic nerves could be interpreted as a centripetal and subsequent centrifugal neural spread of PrP(Sc) for this route of infection. In order to determine whether this propagation is dependent on main components in the axonal cytoskeleton (e.g. neurofilaments, also relevant for the component ;a' of slow axonal transport mechanisms), hamsters were treated -in an additional experiment- with the neurotoxin beta,beta-iminodiproprionitrile (IDPN) around the beginning of the scrapie infection. A comparison of the Western blot signals of PrP(Sc) in the ipsilateral and in the subsequently affected contralateral sciatic nerve segments with the results revealed from IDPN-untreated animals at preclinical and clinical stages of the TSE disease, indicated similar amounts of PrP(Sc). Furthermore, the mean survival time was unchanged in both groups. This in vivo model, therefore, suggests that the propagation of PrP(Sc) along peripheral nerves is not dependent on an intact neurofilament component of the axonal cytoskeleton. Additionally, the model indicates that the spread of PrP(Sc) is not mediated by the slow component ;a' of the axonal transport mechanism.


15-Year experience in the treatment of rectal prolapse in children.
Rectal prolapse is a common and usually self-limited condition in children. Several surgical techniques have been advocated for refractory prolapse. We reviewed our experience with treatment and the outcome of refractory rectal prolapse. Retrospective review was conducted on patients undergoing surgery for rectal prolapse from January 1993 to March 2009. Patients with imperforate anus/cloacal abnormalities, Hirschsprung disease, spina bifida, or prior pull-through were excluded. Twenty patients underwent 23 procedures for rectal prolapse. There were 10 posterior sagittal rectopexies, 6 transabdominal rectopexies, 5 laparoscopic rectopexies, 1 hypertonic saline injection, and 1 anal cerclage. The mean duration of symptoms was 1.6 years (range, 1-10 years). The mean age at operation was 6.8 years (range, 4 months-19 years), with a 5:1 male predominance. There was no operative or perioperative mortality. Median length of follow-up was 7.2 months; 2 patients were lost to follow-up. The overall recurrence rate was 35%. All recurrences followed posterior sagittal rectopexies, which had a 70% recurrence rate. Four patients required reoperation, all done transabdominally (2 open and 2 laparoscopically). None of the 3 remaining patients with mild recurrences required reoperation. A variety of options for management of refractory rectal prolapse in children exist. Laparoscopic rectopexy seems to be safe and a comparatively successful option in these children.


Tunable optical properties of a two-dimensional square-lattice superconductor-dielectric Bragg reflector.
We numerically analyze the optical properties of a two-dimensional (2D) superconducting Bragg reflector (SBR) using the finite element method in conjunction with a two-fluid model. It is found that the wavelength-dependent reflectance spectra of the proposed 2D SBR are strongly dependent on the polarizations of incident light and can be parametrically tuned by the system temperature and the geometric parameters of embedded dielectric rods. Taking advantage of the dispersive superconductor with its zero-refractive index characteristic and the structural periodicity of the proposed superconducting structure, narrow passband filters can be generated near the threshold wavelength. Furthermore, the narrow passband features of the 2D SBR are found to be sustained up to a very large angle of incidence. The extraordinary optical properties imply that the proposed 2D SBR may be applied to the design of an omnidirectional narrowband transmission filter.


Incidence and risk factors of intracranial atherosclerotic stroke: the Northern Manhattan Stroke Study.
To assess the prevalence of risk factors as determinants of intracranial atherosclerosis (IATH)-related stroke in a multi-ethnic community-based cohort. The Northern Manhattan Stroke Study included a population-based incidence study and a nested case-control study. Incident cases of first ischemic stroke were 1:2 when matched to community controls by age, sex, and race/ethnicity. Vascular risk factors were assessed among controls and compared against the following stroke subtypes: IATH, extracranial atherosclerosis (EATH), and non-atherosclerotic (NATH: cardioembolic, lacunar, and cryptogenic). Conditional logistic-regression was used to determine the association between risk factors and stroke subtypes. The crude incidence of IATH was 8/100,000 per year and the relative incidence of IATH was higher than that of EATH in blacks (5.9 vs. 3.2/100,000 per year) and in Hispanics (5.0 vs. 1.7/100,000 per year). The IATH group had a higher prevalence of diabetes mellitus (DM; 67% IATH, 60% EATH, 48% NATH, and 23% controls; p < 0.05 IATH vs. control) and of metabolic syndrome (62% IATH, 40% EATH, 40% NATH, and 35% controls; p < 0.05 IATH vs. control). In multivariate analysis, DM conferred a higher risk for IATH versus NATH stroke (OR, 10.8; 95% CI, 2.0-57 vs. OR, 2.7; 95% CI, 1.9-3.9; p < 0.05) and much lower for EATH (OR, 6.2; 95% CI, 1.2-32). The metabolic syndrome conferred a higher risk for IATH stroke subtype (OR, 4.6; 95% CI, 1.1-18.7) when compared to EATH (OR, 2.3; CI, 0.6-9.1) and NATH (OR, 2.4; CI, 1.7-3.3). DM is a more important determinant for IATH-related stroke than EATH or NATH.


Simplified evaluation of the paradoxical puborectalis contraction with surface electrodes.
Paradoxical puborectalis contraction during defecation is one possible explanation for constipation. The degree of paradoxical contraction can be evaluated by intramuscular electromyography from the puborectalis and external anal sphincter muscles. This study aimed to determine whether a noninvasive technique with surface electrodes placed over the subcutaneous part of the external anal sphincter is feasible in the evaluation of paradoxical activity. Twenty-five patients with constipation were studied. Sphincter muscle activity during strain and squeeze maneuvers was recorded using surface electrodes placed 1 cm from the anal verge. In addition, intramuscular recordings were made simultaneously from the external anal sphincter and puborectalis muscles. The degree of paradoxical activation was calculated as a strain/squeeze index. The patients were examined either in the left lateral position or sitting on a commode. The study revealed significant (P < .01) correlations between indices obtained from the surface anal sphincter recordings and the intramuscular recordings (from the external anal sphincter and the puborectalis muscles). Surface recordings from the external anal sphincter seem to be an equally reliable, less time consuming, and less painful alternative to invasive measurements of paradoxical activity. In a few patients, however, invasive recordings may still be required.


The NFIII/OCT-1 binding site stimulates adenovirus DNA replication in vivo and is functionally redundant with adjacent sequences.
The inverted terminal repeat (ITR) of adenovirus type 5 (Ad5) is 103 bp in length and contains the origin of DNA replication. Cellular transcription factors NFI/CTF and NFIII/OCT-1 bind to sites within the ITR and participate in the initiation of viral DNA replication in vitro. The ITR also contains multiple copies of two conserved sequence motifs that bind the cellular transcription factors SP1 and ATF. We have analyzed a series of viruses that carry deletions at the left terminus of Ad5. A virus carrying a deletion of the NFIII/OCT-1, SP1, and ATF sites within the ITR (mutant dl309-44/107) was wild type for virus growth. However, the deletion of these elements in addition to sequences immediately flanking the ITR (mutant dl309-44/195) resulted in a virus that grew poorly. The analysis of growth parameters of these and other mutants demonstrate that the NFIII/OCT-1 and adjacent SP1 sites augment the accumulation of viral DNA following infection. The function of these elements was most evident in coinfections with a wild-type virus, suggesting that these sites enhance the ability of a limiting trans-acting factor(s), that stimulates viral DNA replication, to interact with the ITR. The results of these analyses indicate functional redundancy between different transcription elements at the left terminus of the Ad5 genome and demonstrate that the NFIII/OCT-1 site and adjacent SP1 site, previously thought to be nonessential for adenovirus growth, play a role in viral DNA replication in vivo.


Validation of a severity-of-illness score in patients with tuberculosis requiring intensive care unit admission.
There is a paucity of data on the determinants of mortality due to tuberculosis (TB) in the intensive care unit (ICU). To develop a simple severity-of-illness score for use in patients with TB admitted to an ICU. A scoring system was generated by retrospectively identifying the four most significant and clinically unrelated predictors of mortality from an existing prospectively collected dataset (January 2012 - May 2013), and combining these with known predictors of poor outcome. Of 83 patients admitted with TB, 38 (45.8%) died in the ICU. The four parameters identified from the retrospective analysis were: (i) HIV co-infection with a CD4cell count <200/µL; (ii) a raised creatinine level: (iii) a chest radiograph showing diffuse parenchymal infiltrates/miliary pattern; and (iv) absence of TB treatment on admission. These were combined with septic shock and a low arterial partial pressure of oxygen/fractional inspired oxygen (P:F) ratio to generate a six-point severity-of-illness score (one point for each parameter). The scores for survivors were significantly lower than those for non-survivors (mean (standard deviation) 2.27 (1.47) v. 3.58 (1.08); p<0.01). A score of ≥2 was associated with significantly higher mortality than a score of <2 (7.1% v. 46.4%; odds ratio (OR) 15.03; 95% confidence interval (CI) 1.86 - 121.32; p<0.01), whereas a score of ≥3 was associated with a significantly higher mortality than a score of <3 (64.6% v. 20.0%; OR 7.29; 95% CI 2.64 - 20.18; p<0.01). The proposed scoring system identified patients at increased risk of dying from TB in the ICU. Further prospective studies are indicated to validate its use.


Primary care practice and cancer suspicion during the first three COVID-19 lockdowns in the UK: a qualitative study.
The COVID-19 pandemic has profoundly affected UK primary care, and as a result the route to cancer diagnosis for many patients. To explore how the pandemic affected primary care practice, in particular cancer suspicion, referral, and diagnosis, and how this experience evolved as the pandemic progressed. Seventeen qualitative interviews were carried out remotely with primary care staff. Staff from practices in England that expressed an interest in trialling an electronic safety-netting tool were invited to participate. Remote, semi-structured interviews were conducted from September 2020 to March 2021. Data analysis followed a thematic analysis and mind-mapping approach. The first lockdown was described as providing time to make adjustments to allow remote and minimal-contact consultations but caused concerns over undetected cancers. These concerns were realised in summer and autumn 2020 as the participants began to see higher rates of late-stage cancer presentation. During the second and third lockdowns patients seemed more willing to consult. This combined with usual winter pressures, demands of the vaccine programme, and surging levels of COVID-19 meant that the third lockdown was the most difficult. New ways of working were seen as positive when they streamlined services but also unsafe if they prevented GPs from accessing all relevant information and resulted in delayed cancer diagnoses. The post-pandemic recovery of cancer care is dependent on the recovery of primary care. The COVID-19 pandemic has highlighted and exacerbated vulnerabilities in primary care but has also provided new ways of working that may help the recovery.


Applying ASTM Standards to Tensile Tests of Musculoskeletal Soft Tissue: Methods to Reduce Grip Failures and Promote Reproducibility.
Tensile testing is an essential experiment to assess the mechanical integrity of musculoskeletal soft tissues, yet standard test methods have not been developed to ensure the quality and reproducibility of these experiments. The ASTM International standards organization has created tensile test standards for common industry materials that specify geometric dimensions of test specimens (coupons) that promote valid failures within the gage section (midsubstance), away from the grips. This study examined whether ASTM test standards for plastics, elastomers, and fiber-reinforced composites are suitable for tensile testing of bovine meniscus along the circumferential fiber direction. We found that dumbbell (DB) shaped coupons based on ASTM standards for elastomers and plastics had an 80% and 60% rate of midsubstance failures, respectively. The rate of midsubstance failures dropped to 20% when using straight (ST) coupons based on ASTM standards for fiber-reinforced composites. The mechanical properties of dumbbell shaped coupons were also significantly greater than straight coupons. Finite element models of the test coupons revealed stress distributions that supported our experimental findings. In addition, we found that a commercial deli-slicer was able to slice meniscus to uniform layer thicknesses that were within ASTM dimensional tolerances. This study provides methods, recommendations, and insights that can advance the standardization of tensile testing in meniscus and other soft fibrous tissues.


Structures of the 2-nitrophenol alkali complexes in solution and the solid state.
The materials studied in this investigation were aqueous solutions (0.02-25.0 mM) of the salts of alkali metal ion (Me(+)) and 2-nitrophenol (2-NP). In the investigation, small-angle X-ray scattering, wide-angle X-ray scattering, and membrane-pressure osmometry were used to study the 2-NP-Me(+) molecular salt structures and the onset of crystallization as a function of concentration and temperature. The experimental methods used to examine the 2-NP-Me(+) molecular salt complexes provided corroborative evidence for the existence of spherical clusters with hydrodynamic diameters between ∼12 Å (Li) and 14 Å (Cs). Guinier plots of the zero-angle scattering peak were characteristic of the scattering from lamellae-like shapes with thicknesses of ∼290 Å. Tetramer and pentamer 2-NP-Me(+) molecular clusters for Me(+) = Li, Na, K, and Rb were assembled from four or five 2-NP molecules bound to a central alkali metal ion. The coordination symmetry around the six coordinated Li(+), Na(+), and K(+) ions was that of a trigonal prism (D3h), with an octahedral arrangement (D2h). The Rb(+) also revealed six-coordinate geometry and the central Rb(+) ion adopted an octahedral arrangement (D2h). The eight-coordinated Cs(+) ions with six 2-NP ligands were characteristic of a square antiprism (D4d). The square antiprism was the outcome of leaving two o-nitro groups and two phenolic oxygens being left intermolecularly uncoordinated to the Cs(+) ion. The 2-NP residues were strictly planar and contained short non-bonded intramolecular distances. van der Waals forces were present between the adjacently stacked phenyl rings. No water molecules were involved as ligands for any of the 2-nitrophenol-Me(+) complexes.


Identification of a novel Sry-related gene and its germ cell-specific expression.
Sox family proteins are characterized by a unique DNA-binding domain, a HMG box which shows at least 50% sequence similarity with mouse Sry, the sex-determining factor. At present almost 30 Sox genes have been identified. Members of this family have been shown to be conserved during evolution and to play key roles during animal development. Some are involved in human diseases, including sex reversal. Here we report the isolation of a novel member of the Sox gene family, Sox30, which may constitute a distinct subgroup of this family. Using a bacterially expressed DNA-binding domain of Sox30, we show that it is able to specifically recognize the ACAAT motif. Furthermore, Sox30 is capable of activating transcription from a synthetic promoter containing the ACAAT motif. The specific expression of Sox30 in normal testes, but not in maturing germ cell-deficient testes, suggests the involvement of Sox30 in differentiation of male germ cells. Mapping analyses revealed that the Sox30 gene is located on human chromosome 5 (5q33) and on mouse chromosome 11.


Consumer willingness to invest money and time for benefits of lifestyle behaviour change: an application of the contingent valuation method.
To use contingent valuation (CV) to derive individual consumer values for both health and broader benefits of a public-health intervention directed at lifestyle behaviour change (LBC) and to examine the feasibility and validity of the method. Participants of a lifestyle intervention trial (n = 515) were invited to complete an online CV survey. Respondents (n = 312) expressed willingness to invest money and time for changes in life expectancy, health-related quality of life (HRQOL) and broader quality of life aspects. Internal validity was tested for by exploring associations between explanatory variables (i.e. income, paid work, experience and risk factors for cardiovascular diseases) and willingness to invest, and by examining ordering effects and respondents' sensitivity to the scope of the benefits. The majority of respondents (94.3%) attached value to benefits of LBC, and 87.4% were willing to invest both money and time. Respondents were willing to invest more for improvements in HRQOL (€42/month; 3 h/week) and broader quality of life aspects (€40/month; 2.6 h/week) than for improvements in life expectancy (€24/month; 2 h/week). Protest answers were limited (3%) and findings regarding internal validity were mixed. The importance of broader quality of life outcomes to consumers suggests that these outcomes are relevant to be considered in the decision making. Our research showed that CV is a feasible method to value both health and broader outcomes of LBC, but generalizability to other areas of public health still needs to be examined. Mixed evidence regarding internal validity pleads for caution to use CV as only the base for decision making.


Suicide and attempted suicide in bipolar disorder: a systematic review of risk factors.
To determine the main risk factors for suicide and nonfatal suicidal behavior in patients with bipolar disorder through a systematic review of the international literature. Studies were identified through electronic searches of MEDLINE (1966-December 2003), EMBASE (1980-December 2003), PsycINFO (1872-November 2003), and Biological Abstracts (1985-December 2003) using index and free-text search terms for bipolar disorder, bipolar depression, manic depression, mania, and affective disorders; combined with terms for self-harm, self-injury, suicide, attempted suicide, automutilation, self-mutilation, self-poisoning, and self-cutting; and combined with terms for risk, case control, cohort, comparative, longitudinal, and follow-up studies. No language restrictions were applied to the search. Included studies were cohort, case-control, and cross-sectional investigations of patients with bipolar disorder in which suicide (13 studies) or attempted suicide (23 studies) was reported as an outcome. The selected studies also used diagnostic tools including the DSM, International Classification of Diseases, and Research Diagnostic Criteria. Meta-analysis of factors reported in more than 1 study identified the main risk factors for suicide as a previous suicide attempt and hopelessness. The main risk factors for nonfatal suicidal behavior included family history of suicide, early onset of bipolar disorder, extent of depressive symptoms, increasing severity of affective episodes, the presence of mixed affective states, rapid cycling, comorbid Axis I disorders, and abuse of alcohol or drugs. Prevention of suicidal behavior in patients with bipolar disorder should include attention to these risk factors in assessment and treatment, including when deciding whether to initiate treatment aimed specifically at reducing suicide risk.


Experimental epothilone B neurotoxicity: results of in vitro and in vivo studies.
Epothilones are a novel class of microtubule-targeting anticancer agents that are neurotoxic. In this study, we investigated the epothilone B toxic effect in vitro and we characterized in vivo the general and neurological side effects of epothilone B administration in Wistar and Fischer rats. The in vitro experiments made it possible to explore a wide concentration range (0.1 nM-1 muM) and evidenced a dose-dependent effect of epothilone B exposure on neuron neurite elongation. This dose-dependent neurotoxic effect was confirmed in both in vivo studies performed on two different rat strains at the neurophysiological, behavioral and pathological levels in the dose range 0.25-1.5 mg/kg iv weekly x 4 weeks and tubulin hyper-polymerization was demonstrated in sciatic nerve specimens. These are the first studies of the neurological effects of epothilone B and they can provide a basis for extending pre-clinical investigation to other members of the epothilone family.


Regulation of the DNA Methylation Landscape in Human Somatic Cell Reprogramming by the miR-29 Family.
Reprogramming to pluripotency after overexpression of OCT4, SOX2, KLF4, and MYC is accompanied by global genomic and epigenomic changes. Histone modification and DNA methylation states in induced pluripotent stem cells (iPSCs) have been shown to be highly similar to embryonic stem cells (ESCs). However, epigenetic differences still exist between iPSCs and ESCs. In particular, aberrant DNA methylation states found in iPSCs are a major concern when using iPSCs in a clinical setting. Thus, it is critical to find factors that regulate DNA methylation states in reprogramming. Here, we found that the miR-29 family is an important epigenetic regulator during human somatic cell reprogramming. Our global DNA methylation and hydroxymethylation analysis shows that DNA demethylation is a major event mediated by miR-29a depletion during early reprogramming, and that iPSCs derived from miR-29a depletion are epigenetically closer to ESCs. Our findings uncover an important miRNA-based approach to generate clinically robust iPSCs.


'The hard work starts now': a glimpse into the lives of carers of community-dwelling stroke survivors.
To explore the personal experiences of carers of stroke survivors and to elicit their views and opinions of what constitute the major issues and concerns of people in their situation. The unexpected nature of stroke can propel people into the role of carer with little or no warning. Some carers of stroke survivors suffer from considerable stress and a range of psychological and physical disorders. A small-scale qualitative study of experienced carers of stroke survivors. Ten carers with experience of caring for a stroke survivor were recruited to the study through community stroke staff. Data were collected using semi-structured interviews and a focus group. The women's main concerns were focused around the need for information and support, including the need for some preparation and training in the necessary skills of caring. To understand fully the lives that carers lead, it is necessary to explore what carers want to do, and expect to do, for the care-recipient. There is a need to develop an understanding of factors that appear to be protective against stress, to identify how they can be enhanced and to develop ways to minimise the impact of those factors that have a detrimental impact on the well-being of carers and on those for whom they care. Health professionals need to ensure that appropriate information is clearly provided for carers. Communication between health professionals to ensure the 'joined up' provision of therapeutic services is another area for continual development. Carers expressed the wish to be better prepared to take on the caring role in the home and would have welcomed some training in this area.


Using active Leptospermum honey in the debridement process: 6 challenging cases from the inner city.
The use of honey-based dressings has been documented for thousands of years. Recent studies suggest their effectiveness may be, in part, related to their ability to facilitate autolytic debridement. Six patients who presented with multiple comorbidities and risk factors for delayed healing whose wounds required debridement were managed with active Leptospermum honey (ALH) to evaluate the safety and effectiveness of this treatment modality. The 6 patients ranged in age from 39 to 81 years. The ALH was covered with a foam dressing; both dressings were changed approximately every 3 days. After 9 to 20 days of use, wounds were completely, or almost completely, debrided, and a 75% concomitant average increase in the amount of granulation tissue in the wound bed was observed. No adverse events were noted. The use of ALH in this case series was effective, and no surgical debridement was needed. Research to compare the efficacy of ALH to other debridement methods is warranted.


Catechol-O-methyltransferase Val158met polymorphism interacts with early experience to predict executive functions in early childhood.
Numerous studies demonstrate that the Methionine variant of the catechol-O-methyltransferase Val158Met polymorphism, which confers less efficient catabolism of catecholamines, is associated with increased focal activation of prefrontal cortex (PFC) and higher levels of executive function abilities. By and large, however, studies of COMT Val158Met have been conducted with adult samples and do not account for the context in which development is occurring. Effects of early adversity on stress response physiology and the inverted U shape relating catecholamine levels to neural activity in PFC indicate the need to take into account early experience when considering relations between genes such as COMT and executive cognitive ability. Consistent with this neurobiology, we find in a prospective longitudinal sample of children and families (N = 1292) that COMT Val158Met interacts with early experience to predict executive function abilities in early childhood. Specifically, the Valine variant of the COMT Val158Met polymorphism, which confers more rather than less efficient catabolism of catecholamines is associated with higher executive function abilities at child ages 48 and 60 months and with faster growth of executive function for children experiencing early adversity, as indexed by cumulative risk factors in the home at child ages 7, 15, 24, and 36 months. Findings indicate the importance of the early environment for the relation between catecholamine genes and developmental outcomes and demonstrate that the genetic moderation of environmental risk is detectable in early childhood.