Quality of Life among Persons with Physical Disability in Udupi Taluk: A Cross Sectional Study.
Disability is not just a health problem or attribute of individuals, but it reflects the problems individuals experience in their interaction with society and physical movements. Disabled persons remain neglected part of society and they also experience various barriers due to restriction of participation. The aim was to assess the quality of life (QOL) of physically disabled persons, the impact of physical disability on activities of daily living (ADL) and to study the awareness about laws and facilities available for disabled persons. A cross-sectional community based study was conducted among 130 physical disabled persons who were selected using convenience sampling technique. The WHO BREF scale was used to assess QOL, while assessment of ADL was done using Barthel Index. Socio-demographic assessment was done using Udai Pareek scale. SPSS version 15.0 was used to analyze data. Categorical variables were expressed as frequencies and percentages. As per guidelines physical, psychological, social, and environmental domain scores for WHO BREF scale were calculated. Among the study participants, 36.2% had a disability from birth that is, congenital. The second common cause of disability was found to be postpolio residual paralysis as it was found among 26.2% respondents. Other causes found were stroke/paralysis and accidents, in 19.2% and 18.5% respondents, respectively. Activities such as transfer, mobility, and stair climbing showed greater impact of physical disability. It was found that 11.5% respondents required help in one of the ADL. QOL score was found to be low under the psychological domain reflecting on negative feelings, bodily image, appearance, spirituality, and self-esteem of respondents. Physical disability had affected social participation as well as marriage of the respondents. However, it was found that very few of them are aware about facilities provided under persons with disability act.


Ancestry informative marker sets for determining continental origin and admixture proportions in common populations in America.
To provide a resource for assessing continental ancestry in a wide variety of genetic studies, we identified, validated, and characterized a set of 128 ancestry informative markers (AIMs). The markers were chosen for informativeness, genome-wide distribution, and genotype reproducibility on two platforms (TaqMan assays and Illumina arrays). We analyzed genotyping data from 825 subjects with diverse ancestry, including European, East Asian, Amerindian, African, South Asian, Mexican, and Puerto Rican. A comprehensive set of 128 AIMs and subsets as small as 24 AIMs are shown to be useful tools for ascertaining the origin of subjects from particular continents, and to correct for population stratification in admixed population sample sets. Our findings provide general guidelines for the application of specific AIM subsets as a resource for wide application. We conclude that investigators can use TaqMan assays for the selected AIMs as a simple and cost efficient tool to control for differences in continental ancestry when conducting association studies in ethnically diverse populations.


Hypersingularity, electromagnetic edge condition, and an analytic hyperbolic wedge model.
It is insufficient to consider that hypersingularity is unphysical solely based on energy considerations. With a proper combination of the two degenerate hypersingular modes, the energy-flux edge condition is satisfied. A hyperbolic wedge model is presented that is much simpler than the previous model for the purpose of studying singular characteristics of the edge fields. This model not only reproduces the sharp edge model as the wedge becomes infinitely sharp but also naturally shows how the two degenerate hypersingular modes of the sharp edge model should be combined. In an incidental study of the effect of rounding edges on numerical computation, I show that the converged results for rounded edges do not converge to a fixed value when the radius of curvature tends to zero, if the corresponding sharp edge supports hypersingularity. I also prove that introducing a small amount of absorption loss for the purpose of improving numerical convergence is effective only when the ratio of the real parts of the permittivities of the two media forming the wedge is close to -1. Finally I remark on the possible illposedness of the hypersingularity problem without imposition of the edge condition.


Clinical impact of dose overestimation by effective path length calculation in stereotactic ablative radiation therapy of lung tumors.
To determine the clinical impact of calculated dose differences between effective path length (EPL) and Monte Carlo (MC) algorithms in stereotactic ablative radiation therapy (SABR) of lung tumors. We retrospectively analyzed the treatment plans and clinical outcomes of 77 consecutive patients treated with SABR for 82 lung tumors between 2003 and 2009 at our institution. Sixty treatments were originally planned using EPL, and 22 using MC. All plans were recalculated for the same beam specifications using MC and EPL, respectively. The doses covering 95%, 50%, and 5% (D95, D50, D5, respectively) of the target volumes were compared between EPL and MC (assumed to be the actual delivered dose), both as physical dose and biologically effective dose. Time to local recurrence was correlated with dose by Cox regression analysis. The relationship between tumor control probability (TCP) and biologically effective dose was determined via logistic regression and used to estimate the TCP decrements due to prescribing by EPL calculations. EPL overestimated dose compared with MC in all tumor dose-volume histogram parameters in all plans. The difference was >10% of the MC D95 to the planning target volume and gross tumor volume in 60 of 82 (73%) and 52 of 82 plans (63%), respectively. Local recurrence occurred in 13 of 82 tumors. Controlling for gross tumor volume, higher physical and biologically effective planning target volume D95 correlated significantly with local control (P = .007 and P = .045, respectively). Compared with MC, prescribing based on EPL translated to a median TCP decrement of 4.3% (range, 1.2%-37%) and a >5% decrement in 46% of tumors. Clinical follow-up for local lung tumor control in a sizable cohort of patients treated with SABR demonstrates that EPL overestimates dose by amounts that substantially decrease TCP in a large proportion. EPL algorithms should be avoided for lung tumor SABR.


Immunological lesions in human uracil DNA glycosylase: association with Bloom syndrome.
Three monoclonal antibodies that react with uracil DNA glycosylase of normal human placenta were tested to determine whether one of the antibodies could be used as a negative marker for Bloom syndrome. As defined by enzyme-linked immunosorbent assay, monoclonal antibody 40.10.09, which reacts with normal human glycosylase, neither recognized nor inhibited native uracil DNA glycosylase from any of five separate Bloom syndrome cell strains. Immunoblot analyses demonstrated that the denatured glycosylase protein from all five Bloom syndrome cell strains was immunoreactive with the 40.10.09 antibody. Further, each native enzyme was immunoreactive with two other anti-human placental uracil DNA glycosylase monoclonal antibodies. In contrast, ELISA reactivity was observed with all three monoclonal antibodies in reactions of glycosylases from 5 normal human cell types and 13 abnormal human cell strains. These results experimentally verify the specificity of the aberrant reactivity of the Bloom syndrome uracil DNA glycosylase. The possibility arises that determination of the lack of immunoreactivity with antibody 40.10.09 may have value in the early diagnosis of Bloom syndrome.


A partial genome assembly of the miniature parasitoid wasp, Megaphragma amalphitanum.
Body size reduction, also known as miniaturization, is an important evolutionary process that affects a number of physiological and phenotypic traits and helps animals conquer new ecological niches. However, this process is poorly understood at the molecular level. Here, we report genomic and transcriptomic features of arguably the smallest known insect-the parasitoid wasp, Megaphragma amalphitanum (Hymenoptera: Trichogrammatidae). In contrast to expectations, we find that the genome and transcriptome sizes of this parasitoid wasp are comparable to other members of the Chalcidoidea superfamily. Moreover, compared to other chalcid wasps the gene content of M. amalphitanum is remarkably conserved. Intriguingly, we observed significant changes in M. amalphitanum transposable element dynamics over time, in which an initial burst was followed by suppression of activity, possibly due to a recent reinforcement of the genome defense machinery. Overall, while the M. amalphitanum genomic data reveal certain features that may be linked to the unusual biological properties of this organism, miniaturization is not associated with a large decrease in genome complexity.


The mTOR inhibitor rapamycin has limited acute anticonvulsant effects in mice.
The mammalian target of rapamycin (mTOR) pathway integrates signals from different nutrient sources, including amino acids and glucose. Compounds that inhibit mTOR kinase activity such as rapamycin and everolimus can suppress seizures in some chronic animal models and in patients with tuberous sclerosis. However, it is not known whether mTOR inhibitors exert acute anticonvulsant effects in addition to their longer term antiepileptogenic effects. To gain insights into how rapamycin suppresses seizures, we investigated the anticonvulsant activity of rapamycin using acute seizure tests in mice. Following intraperitoneal injection of rapamycin, normal four-week-old male NIH Swiss mice were evaluated for susceptibility to a battery of acute seizure tests similar to those currently used to screen potential therapeutics by the US NIH Anticonvulsant Screening Program. To assess the short term effects of rapamycin, mice were seizure tested in ≤ 6 hours of a single dose of rapamycin, and for longer term effects of rapamycin, mice were tested after 3 or more daily doses of rapamycin. The only seizure test where short-term rapamycin treatment protected mice was against tonic hindlimb extension in the MES threshold test, though this protection waned with longer rapamycin treatment. Longer term rapamycin treatment protected against kainic acid-induced seizure activity, but only at late times after seizure onset. Rapamycin was not protective in the 6 Hz or PTZ seizure tests after short or longer rapamycin treatment times. In contrast to other metabolism-based therapies that protect in acute seizure tests, rapamycin has limited acute anticonvulsant effects in normal mice. The efficacy of rapamycin as an acute anticonvulsant agent may be limited. Furthermore, the combined pattern of acute seizure test results places rapamycin in a third category distinct from both fasting and the ketogenic diet, and which is more similar to drugs acting on sodium channels.


Targeted gene silencing to induce permanent sterility.
A non-surgical method to induce sterility would be a useful tool to control feral populations of animals. Our laboratories have experience with approaches aimed at targeting brain cells in vivo with vehicles that deliver a payload of either inhibitory RNAs or genes intended to correct cellular dysfunction. A combination/modification of these methods may provide a useful framework for the design of approaches that can be used to sterilize cats and dogs. For this approach to succeed, it has to meet several conditions: it needs to target a gene essential for fertility. It must involve a method that can selectively silence the gene of interest. It also needs to deliver the silencing agent via a minimally invasive method. Finally, the silencing effect needs to be sustained for many years, so that expansion of the targeted population can be effectively prevented. In this article, we discuss this subject and provide a succinct account of our previous experience with: (i) molecular reagents able to disrupt reproductive cyclicity when delivered to regions of the brain involved in the control of reproduction and (ii) molecular reagents able to ameliorate neuronal disease when delivered systemically using a novel approach of gene therapy.


Breathing with floating ribs: XROMM analysis of lung ventilation in savannah monitor lizards.
The structures and functions of the vertebrate lung and trunk are linked through the act of ventilation, but the connections between these structures and functions are poorly understood. We used X-ray reconstruction of moving morphology (XROMM) to measure rib kinematics during lung ventilation in three savannah monitor lizards (Varanus exanthematicus). All of the dorsal ribs, including the floating ribs, contributed to ventilation; the magnitude and kinematic pattern showed no detectable cranial-to-caudal gradient. The true ribs acted as two rigid bodies connected by flexible cartilage, with the vertebral rib and ventromedial shaft of each sternal rib remaining rigid and the cartilage between them forming a flexible intracostal joint. Rib rotations can be decomposed into bucket handle rotation around a dorsoventral axis, pump handle rotation around a mediolateral axis and caliper motion around a craniocaudal axis. Dorsal rib motion was dominated by roughly equal contributions of bucket and pump rotation in two individuals and by bucket rotation in the third individual. The recruitment of floating ribs during ventilation in monitor lizards is strikingly different from the situation in iguanas, where only the first few true ribs contribute to breathing. This difference may be related to the design of the pulmonary system and life history traits in these two species. Motion of the floating ribs may maximize ventilation of the caudally and ventrolaterally positioned compliant saccular chambers in the lungs of varanids, while restriction of ventilation to a few true ribs may maximize crypsis in iguanas.


SUPPORT tools for evidence-informed policymaking in health 18: Planning monitoring and evaluation of policies.
This article is part of a series written for people responsible for making decisions about health policies and programmes and for those who support these decision makers. The term monitoring is commonly used to describe the process of systematically collecting data to inform policymakers, managers and other stakeholders whether a new policy or programme is being implemented in accordance with their expectations. Indicators are used for monitoring purposes to judge, for example, if objectives are being achieved, or if allocated funds are being spent appropriately. Sometimes the term evaluation is used interchangeably with the term monitoring, but the former usually suggests a stronger focus on the achievement of results. When the term impact evaluation is used, this usually implies that there is a specific attempt to try to determine whether the observed changes in outcomes can be attributed to a particular policy or programme. In this article, we suggest four questions that can be used to guide the monitoring and evaluation of policy or programme options. These are: 1. Is monitoring necessary? 2. What should be measured? 3. Should an impact evaluation be conducted? 4. How should the impact evaluation be done?


Effect of Ala16Val genetic polymorphism of MnSOD on antioxidant capacity and inflammatory response in open heart surgery.
Ischemia-reperfusion injury and inflammation in cardiac surgical patients involves complex humoral and cellular interactions. We investigated the effect of genetic polymorphism of manganase superoxide dismutase (MnSOD) a natural antioxidant, on cytokine release and manganesuperoxide dismutase in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). Forty-two patients undergoing elective CABG with CPB were included in the study. MnSOD polymorphism was performed by polymerase chain reaction (PCR). Levels of interleukin-6 and mangane superoxide dismutase (MnSOD) were measured by enzyme linked immunoabsorbent assay (ELISA). Baseline IL-6 did not differ between patients with different MnSOD genotypes. Postoperatively IL-6 levels were significantly higher in all patients but more significantly in V(VV+AV) carriers (p=0.003). The wild-type AA genotype had the highest preoperative (p<0.05) and postoperative IL-6 level. The MnSOD VV genotype was associated with significantly lower preoperative MnSOD levels compared to the AA carriers (p<0.05). These data demonstrate that MnSOD Ala16Val polymorphism influences IL-6 production and baseline MnSOD activity, suggesting that preoperative MnSOD concentration plays a role in cytokine release.


Indole-3-carbinol and its N-alkoxy derivatives preferentially target ERα-positive breast cancer cells.
Indole-3-carbinol (I3C) is a natural anti-carcinogenic compound found at high concentrations in Brassica vegetables. I3C was recently reported to inhibit neutrophil elastase (NE) activity, while consequently limiting the proteolytic processing of full length cyclin E into pro-tumorigenic low molecular weight cyclin E (LMW-E). In this study, we hypothesized that inhibition of NE activity and resultant LMW-E generation is critical to the anti-tumor effects of I3C. LMW-E was predominately expressed by ERα-negative breast cancer cell lines. However, ERα-positive cell lines demonstrated the greatest sensitivity to the anti-tumor effects of I3C and its more potent N-alkoxy derivatives. We found that I3C was incapable of inhibiting NE activity or the generation of LMW-E. Therefore, this pathway did not contribute to the anti-tumor activity of I3C. Gene expression analyzes identified ligand-activated aryl hydrocarbon receptor (AhR), which mediated sensitivity to the anti-tumor effects of I3C in ERα-positive MCF-7 cells. In this model system, the reactive oxygen species (ROS)-induced upregulation of ATF-3 and pro-apoptotic BH3-only proteins (e.g. NOXA) contributed to the sensitivity of ERα-positive breast cancer cells to the anti-tumor effects of I3C. Overexpression of ERα in MDA-MB-231 cells, which normally lack ERα expression, increased sensitivity to the anti-tumor effects of I3C, demonstrating a direct role for ERα in mediating the sensitivity of breast cancer cell lines to I3C. Our results suggest that ERα signaling amplified the pro-apoptotic effect of I3C-induced AhR signaling in luminal breast cancer cell lines, which was mediated in part through oxidative stress induced upregulation of ATF-3 and downstream BH3-only proteins.


Development and Validation of the Singapore Thyroid Eye Disease Quality of Life Questionnaire.
Current instruments to assess thyroid eye disease (TED) quality of life (QoL) were not developed using modern psychometric theory and may not be applicable to Asian populations. Therefore, we developed a psychometrically robust questionnaire, the Singapore Thyroid Eye Disease Quality of Life questionnaire (STED-QoL), for assessing QoL in Asian patients. This cross-sectional study was conducted at the Singapore National Eye Centre between 2012 and 2015. In Phase 1, content for the questionnaire was developed using qualitative methods. A total of 20 patients participated in three different focus groups. Thematic analysis was conducted to identify relevant themes from which 12 items, rated on a 5-point Likert-type scale, were generated. In Phase 2, the pilot instrument was administered to 59 TED patients and psychometric assessment of the STED-QoL was conducted using Rasch analysis. After collapsing categories from five to four and deleting two misfitting items, we generated a 10-item STED-QoL befitting the Rasch model. The scale showed good criterion validity, with scores decreasing as severity of TED worsened: mild (1.78 logits), moderate (0.27 logits), and severe (0.92 logits). A 'Psychosocial' subscale also had adequate psychometric properties and psychosocial scores were significantly worse in those who underwent surgery for TED compared to those who had not (0.41 vs. 1.82 logits, P = 0.021). The STED-QoL is a robust 10-item questionnaire specifically developed to measure the impact of TED on QoL and psychosocial well-being in an Asian population. QoL assessment is important for holistic management of TED patients.


CD14 expression is increased on monocytes in patients with anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis and correlates with the expression of ANCA autoantigens.
Monocyte subsets with differing functional properties have been defined by their expression of CD14 and CD16. We investigated these subsets in anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis (AAV) and determined their surface expression of ANCA autoantigens. Flow cytometry was performed on blood from 14 patients with active AAV, 46 patients with AAV in remission and 21 controls. The proportion of classical (CD14(high) CD16(neg/low)), intermediate (CD14(high) CD16(high)) and non-classical (CD14(low) CD16(high)) monocytes and surface expression levels of CD14 and CD16 were determined, as well as surface expression of proteinase 3 (PR3) and myeloperoxidase (MPO) on monocyte subsets. There was no change in the proportion of monocytes in each subset in patients with AAV compared with healthy controls. The expression of CD14 on monocytes from patients with active AAV was increased, compared with patients in remission and healthy controls (P < 0.01). Patients with PR3-ANCA disease in remission also had increased monocyte expression of CD14 compared with controls (P < 0.01); however, levels in patients with MPO-ANCA disease in remission were lower than active MPO-ANCA patients, and not significantly different from controls. There was a correlation between CD14 and both PR3 and MPO expression on classical monocytes in AAV patients (r = 0.79, P < 0.0001 and r = 0.42, P < 0.005, respectively). In conclusion, there was an increase in monocyte CD14 expression in active AAV and PR3-ANCA disease in remission. The correlation of CD14 expression with ANCA autoantigen expression in AAV may reflect cell activation, and warrants further investigation into the potential for increased CD14 expression to trigger disease induction or relapse.


Extracellular matrix dynamics in scar-free endometrial repair: perspectives from mouse in vivo and human in vitro studies.
Repair of the postmenstrual endometrium presents a unique opportunity to examine nonscarring repair in an adult tissue. We aimed to characterize and determine the importance of extracellular matrix (ECM) dynamics in cell migration during endometrial repair. Utilizing an in vivo mouse model of postmenstrual repair and an in vitro model of human endometrial re-epithelialization, we determined the dynamic changes in expression of ECM and related factors in both models by array analysis of repairing areas. We also validated expression of integrins, growth factors, protease inhibitors, basement membrane, and adhesion molecules in vitro and in both mouse and human repairing endometrium by quantitative RT-PCR and immunohistochemical studies. Finally, we determined the functional importance of integrin-fibronectin interactions and matrix metalloprotease (MMP)-facilitated cell movement during re-epithelialization and propose a model for cell locomotion during postmenstrual repair. These data demonstrated the dynamic expression and functional importance of ECM interactions in endometrial repair, which may be important for scar-free repair.


Cavity-backed metasurface antennas and their application to frequency diversity imaging.
Frequency diversity antennas with spatially structured radiation patterns reduce the reliance on actively switched elements for beamforming which become increasingly expensive and impractical as frequency increases. As the quality factor Q of a frequency diverse antenna increases, the antenna samples more spatial structure as the number of unique radiated coded spatial patterns correspondingly increases. Antennas that combine hollow cavities and metamaterial apertures achieve both large fractional bandwidth, in excess of 40%, and a high Q of 1600, so that each antenna radiates over 640 unique coded patterns. As compared to switched active antennas, such a passive antenna replaces the 50 antennas and switches that would produce at most (50/2)2=625 unique patterns. Furthermore, the engineered metamaterial apertures enable a radiation efficiency exceeding 60% to be achieved in a single desired polarization. The theory of cavity-backed metasurface antennas is explained, and frequency diverse imaging is demonstrated with a pair of these antennas.


Textures and traction: how tube-dwelling polychaetes get a leg up.
By controlling the traction between its body and the tube wall, a tube-dwelling polychaete can move efficiently from one end of its tube to the other, brace its body during normal functions (e.g., ventilation and feeding), and anchor within its tube avoiding removal by predators. To examine the potential physical interaction between worms and the tubes they live in, scanning electron microscopy was used to reveal and quantify the morphology of worm bodies and the tubes they produce for species representing 13 families of tube-dwelling polychaetes. In the tubes of most species there were macroscopic or nearly macroscopic (∼10 μm-1 mm) bumps or ridges that protruded slightly into the lumen of the tube; these could provide purchase as a worm moves or anchors. At this scale (∼10 μm-1 mm), the surfaces of the chaetal heads that interact with the tube wall were typically small enough to fit within spaces between these bumps (created by the inward projection of exogenous materials incorporated into the tube wall) or ridges (made by secretions on the interior surface of the tube). At a finer scale (0.01-10 μm), there was a second overlap in size, usually between the dentition on the surfaces of chaetae that interact with the tube walls and the texture provided by the secreted strands or microscopic inclusions of the inner linings. These linings had a surprising diversity of micro-textures. The most common micro-texture was a "fabric" of secreted threads, but there were also orderly micro-ridges, wrinkles, and rugose surfaces provided by microorganisms incorporated into the inner tube lining. Understanding the fine structures of tubes in conjunction with the morphologies of the worms that build them gives insight into how tubes are constructed and how worms live within them.


Plasma membrane H-ATPase-dependent citrate exudation from cluster roots of phosphate-deficient white lupin.
White lupin (Lupinus albus L.) is able to grow on soils with sparingly available phosphate (P) by producing specialized structures called cluster roots. To mobilize sparingly soluble P forms in soils, cluster roots release substantial amounts of carboxylates and concomitantly acidify the rhizosphere. The relationship between acidification and carboxylate exudation is still largely unknown. In the present work, we studied the linkage between organic acids (malate and citrate) and proton exudations in cluster roots of P-deficient white lupin. After the illumination started, citrate exudation increased transiently and reached a maximum after 5 h. This effect was accompanied by a strong acidification of the external medium and alkalinization of the cytosol, as evidenced by in vivo nuclear magnetic resonance (NMR) analysis. Fusicoccin, an activator of the plasma membrane (PM) H+-ATPase, stimulated citrate exudation, whereas vanadate, an inhibitor of the H+-ATPase, reduced citrate exudation. The burst of citrate exudation was associated with an increase in expression of the LHA1 PM H+-ATPase gene, an increased amount of H+-ATPase protein, a shift in pH optimum of the enzyme and post-translational modification of an H+-ATPase protein involving binding of activating 14-3-3 protein. Taken together, our results indicate a close link in cluster roots of P-deficient white lupin between the burst of citrate exudation and PM H+-ATPase-catalysed proton efflux.


Evaluation of quality of life and depression levels in individuals with Type 2 diabetes.
Improving the quality of life in diabetic individuals is known to reduce morbidity and mortality. We aimed to investigate the quality of life and depression symptomatology situations and the related factors in patients with Type 2 diabetes mellitus (DM) in this study. In this study, 440 adult patients with Type 2 DM and under treatment admitted to Selcuk University Family Medicine Outpatient Diabetes Education Clinic were included in the study. A questionnaire containing sociodemographic characteristics of the participants, the Short Form 36 (SF-36) quality of life questionnaire was applied with Beck depression inventory face to face interviews. Mean scores of females in all SF-36 subscales were statistically significantly lower than those of male patients. Physical function, physical role limitations, general health, social function, emotional role limitations, and mental health mean scores of the patients with 1-10 years duration of diabetes were found statistically significantly higher than those with 20 years and over duration of diabetes. Physical function, physical role limitations, pain, general health, and social function mean scores in patients using oral antidiabetic drug (OAD) was statistically significantly higher compared to patients using insulin + OAD. The average physical function scores of the patients with no complications were statistically significantly higher than those with two and more complications. Quality of life and depression symptomatology are worse in females, the elderly, the overweight, people with lower level of education, in the widowed or divorced, homemakers, those with low incomes, those with longer duration of diabetes, patients using insulin, and those with two or more complications. There are many medical and sociodemographic factors affecting the quality of life and depressive symptomatology in the individuals with diabetes, so both health care workers and patients should pay the necessary attention to this issue.


Actin and alpha-actinin orchestrate the assembly and maturation of nascent adhesions in a myosin II motor-independent manner.
Using two-colour imaging and high resolution TIRF microscopy, we investigated the assembly and maturation of nascent adhesions in migrating cells. We show that nascent adhesions assemble and are stable within the lamellipodium. The assembly is independent of myosin II but its rate is proportional to the protrusion rate and requires actin polymerization. At the lamellipodium back, the nascent adhesions either disassemble or mature through growth and elongation. Maturation occurs along an alpha-actinin-actin template that elongates centripetally from nascent adhesions. Alpha-Actinin mediates the formation of the template and organization of adhesions associated with actin filaments, suggesting that actin crosslinking has a major role in this process. Adhesion maturation also requires myosin II. Rescue of a myosin IIA knockdown with an actin-bound but motor-inhibited mutant of myosin IIA shows that the actin crosslinking function of myosin II mediates initial adhesion maturation. From these studies, we have developed a model for adhesion assembly that clarifies the relative contributions of myosin II and actin polymerization and organization.


Pilicide ec240 disrupts virulence circuits in uropathogenic Escherichia coli.
Chaperone-usher pathway (CUP) pili are extracellular organelles produced by Gram-negative bacteria that mediate bacterial pathogenesis. Small-molecule inhibitors of CUP pili, termed pilicides, were rationally designed and shown to inhibit type 1 or P piliation. Here, we show that pilicide ec240 decreased the levels of type 1, P, and S piliation. Transcriptomic and proteomic analyses using the cystitis isolate UTI89 revealed that ec240 dysregulated CUP pili and decreased motility. Paradoxically, the transcript levels of P and S pilus genes were increased during growth in ec240, even though the level of P and S piliation decreased. In contrast, the most downregulated transcripts after growth in ec240 were from the type 1 pilus genes. Type 1 pilus expression is controlled by inversion of the fimS promoter element, which can oscillate between phase on and phase off orientations. ec240 induced the fimS phase off orientation, and this effect was necessary for the majority of ec240's inhibition of type 1 piliation. ec240 increased levels of the transcriptional regulators SfaB and PapB, which were shown to induce the fimS promoter phase off orientation. Furthermore, the effect of ec240 on motility was abolished in the absence of the SfaB, PapB, SfaX, and PapX regulators. In contrast to the effects of ec240, deletion of the type 1 pilus operon led to increased S and P piliation and motility. Thus, ec240 dysregulated several uropathogenic Escherichia coli (UPEC) virulence factors through different mechanisms and independent of its effects on type 1 pilus biogenesis and may have potential as an antivirulence compound. CUP pili and flagella play active roles in the pathogenesis of a variety of Gram-negative bacterial infections, including urinary tract infections mediated by UPEC. These are extremely common infections that are often recurrent and increasingly caused by antibiotic-resistant organisms. Preventing piliation and motility through altered regulation and assembly of these important virulence factors could aid in the development of novel therapeutics. This study increases our understanding of the regulation of these virulence factors, providing new avenues by which to target their expression.


Image formation with regard to object shadow for objects inside a scattering medium.
The conventional theory of image transfer through a scattering medium treats objects located upon reflecting surfaces. It is shown that, when an object is located inside a scattering medium and shields a part of space, ignoring the shadowing leads to incorrect results, especially for modern time-gating systems. We develop a general theory of image formation including the shadowing effect when an object is located inside a scattering medium. The example of the observation of a submerged object through a windy ocean surface is chosen to illustrate this theory. A few unexpected effects in imaging of a submerged object are found and discussed, including contrast conversion for a sinking object and higher contrast in the shadow image than in the image of the object itself. The conclusion that using the shadow image for detection of a submerged object can be more efficient than using the image of the object itself is of practical significance.


Apparent total-tract macronutrient digestibility, serum chemistry, urinalysis, and fecal characteristics, metabolites and microbiota of adult dogs fed extruded, mildly cooked, and raw diets1.
Despite their popularity, little research has been performed on lightly cooked and raw diet formats for pets. Therefore, the objective of this study was to determine the apparent total-tract macronutrient digestibility (ATTD); fecal characteristics, metabolites, and microbiota; serum chemistry metabolites; urinalysis; and voluntary physical activity levels of adult dogs fed commercial diets differing in processing type. The diets included: 1) extruded dry kibble (EXT) diet; 2) high-moisture roasted refrigerated (RR) diet; 3) high-moisture grain-free roasted refrigerated (GFRR) diet; and 4) raw (RAW) diet. Eight dogs (mean age = 3.6; mean BW = 13.0 kg) were used in a replicated 4 × 4 Latin square design. Each period consisted of 28 d, with a 14-d adaptation phase followed by a 7-d phase for measuring voluntary physical activity, 1-d adaptation phase to metabolic cages, 5-d phase for fecal and urine collection, and 1 d for blood collection. Except for microbiota, all data were analyzed statistically by mixed models using SAS. Microbiota data were analyzed using Quantitative Insights Into Microbial Ecology (QIIME) and Statistical Analyses of Metagenomic Profiles (STAMP) software. Many differences in digestibility were observed, including greater (P < 0.05) ATTD of CP and fat in dogs fed GFRR and RR than dogs fed EXT. Dogs fed RAW had the lowest fecal pH and DM %, but fecal scores were not affected. Dogs fed RR had higher (P < 0.05) fecal indole and total phenol and indole concentrations than dogs fed the other diets. Dogs fed RAW had a higher (P < 0.05) fecal ammonia concentration than dogs fed the other diets. Fecal microbial diversity was altered by diet, with dogs fed GFRR and RAW having reduced species richness than dogs fed EXT. Dogs fed RR, GFRR, or RAW had lower (P < 0.05) Actinobacteria and higher (P < 0.05) Fusobacteria than dogs fed EXT. Dogs fed RAW or GFRR had higher (P < 0.05) Proteobacteria than dogs fed EXT or RR. Dogs fed RAW had higher (P < 0.05) Bacteroidetes and lower (P < 0.05) Firmicutes than dogs fed EXT. Serum triglycerides were within reference ranges, but greater (P < 0.05) in dogs fed EXT than dogs fed GFRR and RAW. All diets were well tolerated and dogs remained healthy throughout the study. In conclusion, the lightly cooked and raw diets tested were highly palatable, highly digestible, reduced blood triglycerides, maintained fecal quality and serum chemistry, and modified the fecal microbial community of healthy adult dogs.


Early Adaptive Radiation of Birds: Evidence from Fossils from Northeastern China
Late Jurassic and Early Cretaceous birds from northeastern China, including many complete skeletons of Confuciusornis, provide evidence for a fundamental dichotomy in the class Aves that may antedate the temporal occurrence of the Late Jurassic Archaeopteryx. The abundance of Confuciusornis may provide evidence of avian social behavior. Jurassic skeletal remains of an ornithurine bird lend further support to the idea of an early separation of the line that gave rise to modern birds. Chaoyangia, an ornithurine bird from the Early Cretaceous of China, has premaxillary teeth.


Adult T-cell leukemia-lymphoma: current treatment strategies and novel immunological approaches.
Adult T-cell leukemia-lymphoma (ATL) is a peripheral T-cell malignancy, closely associated with human T-cell lymphotropic virus type I infection. Clinically, ATL is classified into four subtypes: acute, lymphoma, chronic and smoldering type. Although the prognosis of chronic and smoldering-type ATL is relatively good, that of patients with acute- or lymphoma-type ATL still remains extremely poor. Zidovudine/IFN-α therapy seems to be promising, although its efficacy has not yet been confirmed in well-designed prospective studies. High-dose chemotherapy with the support of autologous transplantation does not improve outcome. Allogeneic stem cell transplantation is promising and approximately 40% of aggressive ATL patients are expected to survive long-term, although transplantation-related mortality is as high as 40-50%. Stem cell transplantation using reduced-intensity conditioning is also effective and safer, with graft-versus-ATL and graft-versus-human T-cell lymphotropic virus type I effects observed after transplantation. Novel approaches including new agents such as purine nucleoside phosphorylase inhibitors and histone deacetylase inhibitors, or targeted immunotherapy using antichemokine receptor-4 antibody or dendritic cell/peptide vaccine are also warranted.


The prevalence of depressive symptoms among fathers and associated risk factors during the first seven years of their child's life: findings from the Millennium Cohort Study.
Increasing evidence suggests that postnatal paternal depression is associated with adverse emotional, behavioural and cognitive outcomes in children. Despite this, few studies have determined the prevalence of fathers' depressive symptoms during the first few years of their children's lives and explored what factors are related to these symptoms. We estimated the prevalence and examined associated risk factors of paternal depressive symptoms in a nationally representative sample of fathers with children aged between 9 months and 7 years old from the Millennium cohort study. The risk factors examined were maternal depressive symptoms, marital conflict, child temperament, child gender, paternal education, fathers' ethnic background, fathers' employment status, family housing, family income and paternal age. Secondary data analysis was conducted using the UK Millennium cohort study, which consisted of data from England, Scotland, Wales and Northern Ireland of families with infants born in the year 2000/2001. Data from four sweeps were used from when children in the cohort were aged 9 months, 3, 5 and 7 years old (n = 5155-12,396). The prevalence of paternal depressive symptoms over time was 3.6 % at 9 months, 1.2 % at 3 years old, 1.8 % at 5 years and 2.0 % at 7 years (using Kessler cut-off points to categorise high depressive symptoms vs low depressive symptoms). Linear regression trends (using continuous measures of depressive symptoms) indicated that both paternal and maternal depressive symptoms decreased over time, suggesting similar patterns of parents' depressive symptoms after the birth of a child, but the decrease was more evident for mothers. Paternal depressive symptoms were consistently associated with fathers' unemployment, maternal depressive symptoms and marital conflict. Socioeconomic factors such as rented housing when child was 9 months and low family income when child was 5 and 7 years were also associated with higher paternal depressive symptoms. Paternal depressive symptoms decreased among fathers when their children were aged between 9 months to 3 years old. Paternal unemployment, high maternal depressive symptoms and high marital conflict were important risk factors for paternal depressive symptoms. In light of our findings, we would recommend a more family centred approach to interventions for depression in the postnatal period.


Dehydrozingerone protects temozolomide-induced cognitive impairment in normal and C6 glioma rats besides enhancing its anticancer potential.
Considering the cognitive impairment induced by temozolomide (TMZ) in glioblastoma survivors, the present study was aimed to evaluate the protective effect of dehydrozingerone (DHZ) against TMZ-induced cognitive impairment (chemobrain) and C6 cell line-induced glioma in male Wistar rats. In both chemobrain and glioma models, TMZ was administered at a dose of 18 mg/kg i.v every 5th day and DHZ at a dose of 100 mg/kg p.o. daily. Additionally, glioma was induced by intracerebral injection of 5 × 104 C6 rat glioma cells in the cortex in the glioma model. Upon disease induction and treatment with TMZ + DHZ, spatial memory was assessed by the Morris water maze (MWM) test and episodic memory by the novel object recognition test (NORT). The induction of glioma was confirmed by histology of the cortex. Hippocampus and frontal cortex were subjected to antioxidant evaluation. Significant loss of spatial and episodic memory was observed with TMZ treatment which was significantly restored by DHZ. DHZ showed significant improvement in oxidative stress markers reversed the histopathological features in the cortex. TMZ-induced elevation of the glutathione level was also reversed by DHZ, indicating the role of DHZ in the reversal of TMZ resistance. In the glioma model, the improvement in cognition by DHZ correlated with the decrease in tumor volume. Altogether, the study results reveal the role of TMZ in worsening the memory and DHZ in reversing it, besides, improving its anticancer potential.


Genetic prognostic and predictive markers in colorectal cancer.
Despite many studies of the likely survival outcome of individual patients with colorectal cancer, our knowledge of this subject remains poor. Until recently, we had virtually no understanding of individual responses to therapy, but the discovery of the KRAS mutation as a marker of probable failure of epidermal growth factor receptor (EGFR)-targeted therapy is a first step in the tailoring of treatment to the individual. With the application of molecular analyses, as well as the ability to perform high-throughput screens, there has been an explosive increase in the number of markers thought to be associated with prognosis and treatment outcome in this disease. In this Review, we attempt to summarize the sometimes confusing findings, and critically assess those markers already in the public domain.


Visually guided obstacle avoidance in the box jellyfish Tripedalia cystophora and Chiropsella bronzie.
Box jellyfish, cubomedusae, possess an impressive total of 24 eyes of four morphologically different types. Two of these eye types, called the upper and lower lens eyes, are camera-type eyes with spherical fish-like lenses. Compared with other cnidarians, cubomedusae also have an elaborate behavioral repertoire, which seems to be predominantly visually guided. Still, positive phototaxis is the only behavior described so far that is likely to be correlated with the eyes. We have explored the obstacle avoidance response of the Caribbean species Tripedalia cystophora and the Australian species Chiropsella bronzie in a flow chamber. Our results show that obstacle avoidance is visually guided. Avoidance behavior is triggered when the obstacle takes up a certain angle in the visual field. The results do not allow conclusions on whether color vision is involved but the strength of the response had a tendency to follow the intensity contrast between the obstacle and the surroundings (chamber walls). In the flow chamber Tripedalia cystophora displayed a stronger obstacle avoidance response than Chiropsella bronzie since they had less contact with the obstacles. This seems to follow differences in their habitats.


Novel immunoassays for detection of CUZD1 autoantibodies in serum of patients with inflammatory bowel diseases.
Pancreatic autoantibodies (PABs) are detected in patients with inflammatory bowel disease (IBD). Their prevalence is higher in Crohn's disease (CrD) than in ulcerative colitis (UC). Glycoprotein 2 (GP2) and, more recently, CUB and zona pellucida-like domain-containing protein 1 (CUZD1) have been identified as target autoantigens of PAB. The clinical utility of CUZD1 autoantibodies has only recently been assessed by indirect immunofluorescence (IIF) assays. In this study, we developed and validated novel immunoassays for the detection of CUZD1 autoantibodies. Recombinant CUZD1 protein was utilized as a solid-phase antigen for the development of two immunoassays for the detection of IgG and IgA CUZD1 autoantibodies. Serum samples from 100 patients with CrD, 100 patients with UC, 129 patients assessed for various autoimmune diseases (vADs) and 50 control individuals were analyzed. Two immunofluorometric assays for the detection of IgG and IgA CUZD1-specific antibodies were developed. CUZD1 autoantibodies were detected in 12.5% (25/200) IBD patients, including 16% of patients with CrD and in 9% of patients with UC (CrD vs. UC, p<0.05), compared with 3.1% (4/129) patients suspected of having vADs (CrD vs. ADs, p<0.05; UC vs. ADs, p=0.08). CUZD1 autoantibody positivity was not found to be related to disease location, age of disease onset or disease phenotype. This is the first study to describe novel IgA and IgG CUZD1 autoantibody enzyme-linked immunosorbent assay. These immunoassays agree well with standard IIF techniques and can be utilized in multicenter studies to investigate the diagnostic and clinical utility of CUZD1 autoantibodies.


Oral Ondansetron Administration to Dehydrated Children in Pakistan: A Randomized Clinical Trial.
Ondansetron is an effective antiemetic employed to prevent vomiting in children with gastroenteritis in high-income countries; data from low- and middle-income countries are sparse. We conducted a randomized, double-blind, placebo-controlled superiority trial in 2 pediatric emergency departments in Pakistan. Dehydrated children aged 6 to 60 months with ≥1 diarrheal (ie, loose or liquid) stool and ≥1 vomiting episode within the preceding 4 hours were eligible to participate. Participants received a single weight-based dose of oral ondansetron (8-15 kg: 2 mg; >15 kg: 4 mg) or identical placebo. The primary outcome was intravenous administration of ≥20 mL/kg over 4 hours of an isotonic fluid within 72 hours of random assignment. All 918 (100%) randomly assigned children completed follow-up. Intravenous rehydration was administered to 14.7% (68 of 462) and 19.5% (89 of 456) of those administered ondansetron and placebo, respectively (difference: -4.8%; 95% confidence interval [CI], -9.7% to 0.0%). In multivariable logistic regression analysis adjusted for other antiemetic agents, antibiotics, zinc, and the number of vomiting episodes in the preceding 24 hours, children administered ondansetron had lower odds of the primary outcome (odds ratio: 0.70; 95% CI, 0.49 to 1.00). Fewer children in the ondansetron, relative to the placebo group vomited during the observation period (difference: -12.9%; 95% CI, -18.0% to -7.8%). The median number of vomiting episodes (P < .001) was lower in the ondansetron group. Among children with gastroenteritis-associated vomiting and dehydration, oral ondansetron administration reduced vomiting and intravenous rehydration use. Ondansetron use may be considered to promote oral rehydration therapy success among dehydrated children in low- and middle-income countries.


The role of core groups in the emergence and dissemination of antimicrobial-resistant N gonorrhoeae.
Core groups contain individuals who are regularly infected with Neisseria gonorrhoeae and are able to transmit their infection to a large number of sexual partners. Classical core groups, such as sex workers and their male partners, or men who have sex with men (MSM), have contributed to the emergence and spread of antimicrobial-resistant N gonorrhoeae over many years. Sex workers and their clients were the most important core group driving the dissemination of penicillinase-producing N gonorrhoeae in the 1970s. Such individuals have continued to contribute to penicillinase-producing N gonorrhoeae outbreaks as well as to the subsequent emergence of gonococcal resistance to fluoroquinolones, macrolides, spectinomycin and cephalosporins in various settings. MSM have been a very important core group since the 1980s, first with the spread of TetM-expressing N gonorrhoeae and, second, with the dissemination of fluoroquinolone-resistant N gonorrhoeae. MSM-associated sexual networks have most recently been critical to the spread of gonococci resistant to third generation extended spectrum cephalosporins, including cefixime and ceftriaxone. Individuals within other core groups have also been linked to the transmission of antimicrobial-resistant gonorrhoea, such as military personnel, travellers, drug users, young adults, older men and members of street gangs. Understanding core behaviours and their geospatial clustering is essential for an optimal public health response to the rising prevalence of antimicrobial-resistant gonorrhoea. Furthermore, rapid and effective treatment of N gonorrhoeae infections in core individuals and their sexual partners should be a priority for gonorrhoea control programmes.


Antibody-based detection of ERG rearrangement-positive prostate cancer.
TMPRSS2-ERG gene fusions occur in 50% of prostate cancers and result in the overexpression of a chimeric fusion transcript that encodes a truncated ERG product. Previous attempts to detect truncated ERG products have been hindered by a lack of specific antibodies. Here, we characterize a rabbit anti-ERG monoclonal antibody (clone EPR 3864; Epitomics, Burlingame, CA) using immunoblot analysis on prostate cancer cell lines, synthetic TMPRSS2-ERG constructs, chromatin immunoprecipitation, and immunofluorescence. We correlated ERG protein expression with the presence of ERG gene rearrangements in prostate cancer tissues using a combined immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) analysis. We independently evaluated two patient cohorts and observed ERG expression confined to prostate cancer cells and high-grade prostatic intraepithelial neoplasia associated with ERG-positive cancer, as well as vessels and lymphocytes (where ERG has a known biologic role). Image analysis of 131 cases demonstrated nearly 100% sensitivity for detecting ERG rearrangement prostate cancer, with only 2 (1.5%) of 131 cases demonstrating strong ERG protein expression without any known ERG gene fusion. The combined pathology evaluation of 207 patient tumors for ERG protein expression had 95.7% sensitivity and 96.5% specificity for determining ERG rearrangement prostate cancer. In conclusion, this study qualifies a specific anti-ERG antibody and demonstrates exquisite association between ERG gene rearrangement and truncated ERG protein product expression. Given the ease of performing IHC versus FISH, ERG protein expression may be useful for molecularly subtyping prostate cancer based on ERG rearrangement status and suggests clinical utility in prostate needle biopsy evaluation.


Subtypes of minimal residual disease, association with Gleason score, risk and time to biochemical failure in pT2 prostate cancer treated with radical prostatectomy.
The Gleason score is a strong prognostic factor for treatment failure in pathologically organ-confined prostate cancer (pT2) treated by radical prostatectomy (RP). However, within each Gleason score, there is clinical heterogeneity with respect to treatment outcome, even in patients with the same pathological stage and prostate-specific antigen (PSA) at diagnosis. This may be due to minimal residual disease (MRD) remaining after surgery. We hypothesise that the sub-type of MRD determines the risk of and timing of treatment failure, is a biological classification, and may explain in part clinical heterogeneity. We present a study of pT2 patients treated with RP, the subtypes of MRD for each Gleason score and clinical outcomes. Patients with Gleason ≤6 (G6) or Gleason 7 (G7) pT2 cancer participated in the study. One month after surgery, blood was taken for circulating prostate cell (CPCs); mononuclear cells were obtained by differential gel centrifugation and identified using immunocytochemistry with anti-PSA. The detection of one CPC/sample was defined as a positive test. Touch-preparations from bone-marrow biopsies were used to detect micro-metastasis using immunocytochemistry with anti-PSA. Biochemical failure was defined as a PSA >0.2 ng/mL. Patients were classified as: Group A MRD negative (CPC and micro-metastasis negative), Group B (only micro-metastasis positive) and Group C (CPC positive). Biochemical failure-free survival (BFFS) using Kaplan-Meier and time to failure using Restricted Mean Survival Time (RMST) after 10 years of follow-up were calculated for each group based on the Gleason score. Of a cohort of 253 men, four were excluded for having Gleason 8 or 9 prostate cancer, leaving a study group of 249 men of whom 52 had G7 prostate cancer. G7 patients had a higher frequency of MRD (69% versus 36%) and worse prognosis. G6 and G7 patients negative for MRD had similar BBFS rates, 98% at 10 years, time to failure 9.9 years. Group C, G6 patients had a higher BFFS and longer time to failure compared to G7 patients (19% versus 5% and 7 versus 3 years). Group B showed similar results up to 5 years, thereafter G6 had a lower BFFS 63% versus 90%. G7 and G6 pT2 patients have different patterns of MRD and relapse. Risk stratification using MRD sub-types may help to define the need for adjuvant therapy. This needs confirmation with large randomised long-term trials.


An elevated body mass index does not reduce survival after esophagectomy for cancer.
Incidences of esophageal cancer and obesity are both rising in the United States. The aim of this study was to determine the influence of elevated body mass index on outcomes after esophagectomy for cancer. Overall and disease-free survivals in obese (BMI ≥ 30), overweight (BMI 25-29), and normal-weight (BMI 20-24) patients undergoing esophagectomy constituted the study end points. Survivals were calculated by the Kaplan-Meier method, and differences were analyzed by log rank method. The study included 166 obese, 176 overweight, and 148 normal-weight patients. These three groups were similar in terms of demographics and comorbidities, with the exception of younger age (62.5 vs. 66.2 vs. 65.3 years, P = 0.002), and higher incidence of diabetes (23.5 vs. 11.4 vs. 10.1%, P = 0.001) and hiatal hernia (28.3 vs. 14.8 vs. 20.3%, P = 0.01) in obese patients. Rates of adenocarcinoma histology were higher in obese patients (90.8 vs. 90.9 vs. 82.5%, P = 0.03). Despite similar preoperative stage, obese patients were less likely to receive neoadjuvant treatment (47.6 vs. 54.5 vs. 66.2%, P = 0.004). Response to neoadjuvant treatment, type of surgery performed, extent of lymphadenectomy, rate of R0 resections, perioperative complications, and administration of adjuvant chemotherapy were not influenced by BMI. At a median follow-up of 25 months, 5-year overall and disease-free survivals were longer in obese patients (respectively, 48, 41, 34%, P = 0.01 and 48, 44, 34%, P = 0.01). In our experience, an elevated BMI did not reduce overall and disease-free survivals after esophagectomy for cancer.


Crystal structure of the extracellular region of human tissue factor.
Tissue factor is a cell-surface glycoprotein receptor which initiates the blood coagulation cascade after vessel injury by interacting with blood clotting factor VII/VIIa and which is implicated in various pathological processes. When bound to tissue factor, factor VII is readily converted to the active protease factor VIIa by trace amounts of factors Xa, IXa or VIIa. Human tissue factor consists of 263 residues, the first 219 of which comprise the extracellular region. We have determined the crystal structure of the extracellular region at a resolution of 2.2 A. Tissue factor consists of two immunoglobulin-like domains associated through an extensive, novel, interdomain interface region. The binding site for factor VII lies at the interface region and involves residues from domain 1 and an extended loop (binding 'finger') of domain 2. This is the first reported structure of a representative of the class 2 cytokine receptor family, which also includes interferon-alpha, interferon-gamma (refs 2, 3) and interleukin-10 (ref. 4) receptors.


A comparative review of HLA associations with hepatitis B and C viral infections across global populations.
Hepatitis B (HBV) and hepatitis C (HCV) viral infection or co-infection leads to risk of development of chronic infection, cirrhosis and hepatocellular carcinoma (HCC). Immigration and globalization have added to the challenges of public health concerns regarding chronic HBV and HCV infections worldwide. The aim of this study is to review existing global literature across ethnic populations on HBV and HCV related human leukocyte antigen (HLA) associations in relation to susceptibility, viral persistence and treatment. Extensive literature search was conducted to explore the HLA associations in HBV and HCV infections reported across global populations over the past decade to understand the knowledge status, weaknesses and strengths of this information in different ethnic populations. HLA DR13 is consistently associated with HBV clearance globally. HLADRB1*11/*12 alleles and DQB1*0301 are associated with HBV persistence but with HCV clearance worldwide. Consistent association of DRB1*03 and *07 is observed with HCV susceptibility and non-responsiveness to HBV vaccination across the population. HLA DR13 is protective for vertical HBV and HCV transmission in Chinese and Italian neonates, but different alleles are associated with their susceptibility in these populations. HLA class I molecule interactions with Killer cell immunoglobulin like receptors (KIR) of natural killer (NK) cells modulate HCV infection outcome via regulating immune regulatory cells and molecules. HLA associations with HBV vaccination, interferon therapy in HBV and HCV, and with extra hepatic manifestations of viral hepatitis are also discussed. Systematic studies in compliance with global regulatory standards are required to identify the HLA specific viral epitope, stage specific T cell populations interacting with different HLA alleles during disease progression and viral clearance of chronic HBV or HCV infections among different ethnic populations. These studies would facilitate stage specific therapeutic strategies for clearance of HBV and HCV infections or co-infections across global populations and aid in identification of HBV-HCV combined vaccine. HLA associations of chronic HBV or HCV development with confounding host factors including alcohol, drug abuse, insulin resistance, age and gender are lacking and warrant detailed investigation across global populations.


Does Science Advance One Funeral at a Time?
We examine how the premature death of eminent life scientists alters the vitality of their fields. While the flow of articles by collaborators into affected fields decreases after the death of a star scientist, the flow of articles by non-collaborators increases markedly. This surge in contributions from outsiders draws upon a different scientific corpus and is disproportionately likely to be highly cited. While outsiders appear reluctant to challenge leadership within a field when the star is alive, the loss of a luminary provides an opportunity for fields to evolve in new directions that advance the frontier of knowledge within them.


Therapeutic advances in leukemia and myelodysplastic syndrome over the past 40 years.
Major therapeutic progress has been accomplished in leukemia and myelodysplastic syndrome (MDS) over the past 40 years, which may not be fully appreciated by the larger medical community. The objective of this review was to briefly highlight the treatment breakthroughs in leukemia and MDS. Therapeutic progress happened through better understanding of disease pathophysiologies and rational development of targeted agents, like imatinib mesylate in chronic myeloid leukemia (CML), and through astute, empirical discoveries in the clinic, like all-trans retinoic acid and arsenic trioxide in acute promyelocytic leukemia (APL) and chlorodeoxyadenosine in hairy cell leukemia (HCL). Today, the 5- to 10-year survival rates in patients with APL and HCL exceed 80%. In patients with CML, imatinib therapy has been associated with estimated 5- to 7-year survival rates from 85% to 90%. In patients with adult acute lymphocytic leukemia, modern intensive regimens have improved the 5-year survival rates from 20% up to 40%. In patients with chronic lymphocytic leukemia, chemoimmunotherapy recently produced high rates of quality responses and improved long-term outcome. In younger patients with acute myeloid leukemia (AML), the 5-year survival rates range from 40% to 50%, although elderly AML remains a therapeutic challenge. In patients with MDS, it was recently demonstrated that epigenetic therapy with hypomethylating agents improved survival. Much therapeutic progress has been witnessed in leukemia and MDS, and much more is expected to occur soon.


Galectin-9 prolongs the survival of septic mice by expanding Tim-3-expressing natural killer T cells and PDCA-1+ CD11c+ macrophages.
Galectin-9 ameliorates various inflammatory conditions including autoimmune diseases by regulating T cell and macrophage/dendritic cell (DC) functions. However, the effect of galectin-9 on polymicrobial sepsis has not been assessed. We induced polymicrobial sepsis by cecal ligation and puncture (CLP) in mice. The survival rate was compared between galectin-9- and PBS-treated CLP mice. An ELISA was used to compare the levels of various cytokines in the plasma and culture supernatants. Fluorescence-activated cell sorting analysis was further performed to compare the frequencies of subpopulations of spleen cells. Galectin-9 exhibited a protective effect in polymicrobial sepsis as demonstrated in galetin-9 transgenic mice and therapeutic galectin-9 administration. In contrast, such effect was not observed in nude mice, indicating the involvement of T cells in galectin-9-mediated survival prolongation. Galectin-9 decreased TNFα, IL-6, IL-10 and, high mobility group box 1 (HMGB1) and increased IL-15 and IL-17 plasma and spleen levels. Galectin-9 increased the frequencies of natural killer T (NKT) cells and PDCA-1+ CD11c+ macrophages (pDC-like macrophages) but did not change the frequency of CD4 or CD8 T cells, γδT cells or conventional DC. As expected, galectin-9 decreased the frequency of Tim-3+ CD4 T cells, most likely Th1 and Th17 cells. Intriguingly, many spleen NK1.1+ NKT cells and pDC-like macrophages expressed Tim-3. Galectin-9 increased the frequency of Tim-3-expressing NK1.1+ NKT cells and pDC-like macrophages. Galectin-9 further increased IL-17+ NK1.1+ NKT cells. These data suggest that galectin-9 exerts therapeutic effects on polymicrobial sepsis, possibly by expanding NKT cells and pDC-like macrophages and by modulating the production of early and late proinflammatory cytokines.


Safety of Osteopathic Cranial Manipulative Medicine as an Adjunct to Conventional Postconcussion Symptom Management: A Pilot Study.
Osteopathic cranial manipulative medicine is not a well-established adjunct to conventional treatment for patients with postconcussion symptoms. To determine whether adjunctive osteopathic cranial manipulative medicine is safe for patients with concussion when accompanied by conventional treatments. Prospective observational pilot study. Outpatient concussion clinic. Patients who sustained a concussion were prospectively recruited from an outpatient concussion clinic by a neuropsychologist specialized in concussion. All participants were identified to have a cranial dysfunction. Each eligible participant received 1 session of the osteopathic cranial manipulative medicine vault hold technique. Self-reported adverse events during or after 1 session of the osteopathic cranial manipulative medicine procedure and improvement in concussion symptoms at return to follow-up. None of the 9 participants reported adverse events during or immediately after receiving osteopathic cranial manipulative medicine. Five of the 7 participants who returned for follow-up demonstrated improvement in their overall concussion symptoms based on the Post-Concussion Symptom Scale scores. Osteopathic cranial manipulative medicine was considered a safe adjunctive treatment option to improve concussion-related symptoms and recovery.


Higher concentrations of nanoscale zero-valent iron (nZVI) in soil induced rice chlorosis due to inhibited active iron transportation.
In this study, the effects of concentrations 0, 100, 250, 500, 750 and 1000 mg kg(-1) of nanoscale zero-valent iron (nZVI) on germination, seedlings growth, physiology and toxicity mechanisms were investigated. The results showed that nZVI had no effect on germination, but inhibited the rice seedlings growth in higher concentrations (>500 mg kg(-1) nZVI). The highest suppression rate of the length of roots and shoots reached 46.9% and 57.5%, respectively. The 1000mg kg(-1) nZVI caused the highest suppression rates for chlorophyll and carotenoids, at 91.6% and 85.2%, respectively. In addition, the activity of antioxidant enzymes was altered by the translocation of nanoparticles and changes in active iron content. Visible symptoms of iron deficiency were observed at higher concentrations, at which the active iron content decreased 61.02% in the shoots, but the active iron content not decreased in roots. Interestingly, the total and available amounts of iron in the soil were not less than those in the control. Therefore, the plants iron deficiency was not caused by (i) deficiency of available iron in the soil and (ii) restraint of the absorption that plant takes in the available iron, while induced by (ⅲ) the transport of active iron from the root to the shoot was blocked. The cortex tissues were seriously damaged by nZVI which was transported from soil to the root, these were proved by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS). This current study shows that the mechanism of iron deficiency in rice seedling was due to transport of active iron from the root to the shoot blocked, which was caused by the uptake of nZVI.


Molecular epidemiology of malaria in Yaounde, Cameroon I. Analysis of point mutations in the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum.
Resistance to antifolate antimalarial drugs (cycloguanil, a biologically active metabolite of proguanil, and pyrimethamine) is associated with a Ser- to Asn-108 point mutation in the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum. The frequency of this mutation was studied in 127 clinical isolates obtained in Yaounde, Cameroon using a simple and rapid molecular technique based on the polymerase chain reaction and restriction fragment length polymorphism. Of the 127 isolates, pure wild-type Ser-108 codon, pure mutant-type Asn-108 codon, and mixed codons were observed in 66, 55, and six parasites, respectively. The proportion of antifolate-resistant, pure mutant-type codon, with respect to pure wild-type or mixed alleles, was 43% (55 of 127). The results of the molecular assay were compared with those of semimicro isotopic in vitro assay in 34 isolates. All 17 pure Ser-108 isolates and two isolates with mixed alleles were sensitive to both pyrimethamine (50% inhibitory concentration [IC50] < 100 nM) and cycloguanil (IC50 < 50 nM). Fourteen of 15 isolates with the mutant-type Asn-108 codon were resistant to pyrimethamine and cycloguanil. One isolate with Asn-108 showed a slightly elevated pyrimethamine IC50 (78 nM), which was within the sensitive range. This study provides further evidence that antifolate-resistant P. falciparum isolates are already present in Yaounde, Cameroon.


Meta-analysis: melatonin for the treatment of primary sleep disorders.
To investigate the efficacy of melatonin compared to placebo in improving sleep parameters in patients with primary sleep disorders. PubMed was searched for randomized, placebo-controlled trials examining the effects of melatonin for the treatment of primary sleep disorders. Primary outcomes examined were improvement in sleep latency, sleep quality and total sleep time. Meta-regression was performed to examine the influence of dose and duration of melatonin on reported efficacy. Adults and children diagnosed with primary sleep disorders. Melatonin compared to placebo. Nineteen studies involving 1683 subjects were included in this meta-analysis. Melatonin demonstrated significant efficacy in reducing sleep latency (weighted mean difference (WMD) = 7.06 minutes [95% CI 4.37 to 9.75], Z = 5.15, p<0.001) and increasing total sleep time (WMD = 8.25 minutes [95% CI 1.74 to 14.75], Z = 2.48, p = 0.013). Trials with longer duration and using higher doses of melatonin demonstrated greater effects on decreasing sleep latency and increasing total sleep time. Overall sleep quality was significantly improved in subjects taking melatonin (standardized mean difference = 0.22 [95% CI: 0.12 to 0.32], Z = 4.52, p<0.001) compared to placebo. No significant effects of trial duration and melatonin dose were observed on sleep quality. This meta-analysis demonstrates that melatonin decreases sleep onset latency, increases total sleep time and improves overall sleep quality. The effects of melatonin on sleep are modest but do not appear to dissipate with continued melatonin use. Although the absolute benefit of melatonin compared to placebo is smaller than other pharmacological treatments for insomnia, melatonin may have a role in the treatment of insomnia given its relatively benign side-effect profile compared to these agents.


p53 and APC mutations are detectable in the plasma and serum of patients with colorectal cancer (CRC) or adenomas.
A variety of tumor-related DNA has been detected in the plasma and serum of cancer patients, including RAS, the BCL2-IgH transgene, and p53. It is not known whether the APC gene, which is frequently mutated in colorectal cancer (CRC), can be identified in the plasma of CRC patients. Similarly, it is unknown whether another tumor suppressor gene altered in CRC, p53, is detectable in people with precursor lesions to CRC. The plasma of 240 colonoscopy patients was tested for mutations at two frequently altered sites (codons 175 and 248). A restriction enzyme-mediated enrichment assay was used to detect these mutants. We also tested tumor tissue from 8 patients with CRC for mutations in the mutation cluster region of the APC gene using direct DNA sequencing. Corresponding plasma from any positive patient was then similarly sequenced. Three plasma p53 mutations were identified. Two of these patients had adenomas at biopsy, and 1 had a hyperplastic polyp. All were tested for the same p53 mutations, and 1 of the adenomas was positive. One of the 8 CRC patients had a 5-base-pair deletion in the cancer and the same deletion was detectable in that patient's plasma, although at an amount that we estimate at 1-5% of the total APC DNA present. APC mutations are detectable in the plasma of CRC patients. p53 mutants are detectable in the plasma of colorectal adenoma patients. These may have important implications for cancer screening and diagnosis in the large intestine and suggest that all malignant and premalignant DNA alterations from the colon are identifiable in the blood.


Plasmid segregation: how to survive as an extra piece of DNA.
Non-essential extra-chromosomal DNA elements such as plasmids are responsible for their own propagation in dividing host cells, and one means to ensure this is to carry a miniature active segregation system reminiscent of the mitotic spindle. Plasmids that are maintained at low numbers in prokaryotic cells have developed a range of such active partitioning systems, which are characterized by an impressive simplicity and efficiency and which are united by the use of dynamic, nucleotide-driven filaments to separate and position DNA molecules. A comparison of different plasmid segregation systems reveals (i) how unrelated filament-forming and DNA-binding proteins have been adopted and modified to create a range of simple DNA segregating complexes and (ii) how subtle changes in the few components of these DNA segregation machines has led to a remarkable diversity in the molecular mechanisms of closely related segregation systems. Here, our current understanding of plasmid segregation systems is reviewed and compared with other DNA segregation systems, and this is extended by a discussion of basic principles of plasmid segregation systems, evolutionary implications and the relationship between an autonomous DNA element and its host cell.


Functional tooth restoration by next-generation bio-hybrid implant as a bio-hybrid artificial organ replacement therapy.
Bio-hybrid artificial organs are an attractive concept to restore organ function through precise biological cooperation with surrounding tissues in vivo. However, in bio-hybrid artificial organs, an artificial organ with fibrous connective tissues, including muscles, tendons and ligaments, has not been developed. Here, we have enveloped with embryonic dental follicle tissue around a HA-coated dental implant, and transplanted into the lower first molar region of a murine tooth-loss model. We successfully developed a novel fibrous connected tooth implant using a HA-coated dental implant and dental follicle stem cells as a bio-hybrid organ. This bio-hybrid implant restored physiological functions, including bone remodelling, regeneration of severe bone-defect and responsiveness to noxious stimuli, through regeneration with periodontal tissues, such as periodontal ligament and cementum. Thus, this study represents the potential for a next-generation bio-hybrid implant for tooth loss as a future bio-hybrid artificial organ replacement therapy.


Neuroinflammation-Induced Interactions between Protease-Activated Receptor 1 and Proprotein Convertases in HIV-Associated Neurocognitive Disorder.
The proprotein convertases (PCs) furin, PC5, PACE4, and PC7 cleave secretory proteins after basic residues, including the HIV envelope glycoprotein (gp160) and Vpr. We evaluated the abundance of PC mRNAs in postmortem brains of individuals exhibiting HIV-associated neurocognitive disorder (HAND), likely driven by neuroinflammation and neurotoxic HIV proteins (e.g., envelope and Vpr). Concomitant with increased inflammation-related gene expression (interleukin-1β [IL-1β]), the mRNA levels of the above PCs are significantly increased, together with those of the proteinase-activated receptor 1 (PAR1), an inflammation-associated receptor that is cleaved by thrombin at ProArg41↓ (where the down arrow indicates the cleavage location), and potentially by PCs at Arg41XXXXArg46↓. The latter motif in PAR1, but not its R46A mutant, drives its interactions with PCs. Indeed, PAR1 upregulation leads to the inhibition of membrane-bound furin, PC5B, and PC7 and inhibits gp160 processing and HIV infectivity. Additionally, a proximity ligation assay revealed that furin and PC7 interact with PAR1. Reciprocally, increased furin expression reduces the plasma membrane abundance of PAR1 by trapping it in the trans-Golgi network. Furthermore, soluble PC5A/PACE4 can target/disarm cell surface PAR1 through cleavage at Arg46↓. PACE4/PC5A decreased calcium mobilization induced by thrombin stimulation. Our data reveal a new PC-PAR1-interaction pathway, which offsets the effects of HIV-induced neuroinflammation, viral infection, and potentially the development of HAND.


Perspective on the human cough reflex.
This review dissects the complex human cough reflex and suggests hypotheses about the evolutionary basis for the reflex. A mechanosensory-induced cough reflex conveys through branches of myelinated Aδ nerve fibers is not chemically reactive (i.e., capsaicin, bradykinin); possibly, its evolution is to prevent the harmful effects of aspiration of gastric or particulate contents into the lungs. This became necessary as the larynx moves closer to the opening of the esophagus as human ancestors adapt phonation over olfaction beginning less than 10 million years ago. The second type of cough reflex, a chemosensory type, is carried by unmyelinated C fibers. Supposedly, its origin dates back when prehistoric humans began living in close proximity to each other and were at risk for infectious respiratory diseases or irritant-induced lung injury. The mechanism for the latter type of cough is analogous to induced pain after tissue injury; and, it is controlled by the identical transient receptor potential vanilloid cation channel (TRPV1). The airways do not normally manifest nociceptive pain from a stimulus but the only consistent response that capsaicin and lung inflammation provoke in healthy human airways is cough. TRPA1, another excitatory ion channel, has been referred to as the "irritant receptor" and its activation also induces cough. For both types of cough, the motor responses are identical and via coordinated, precisely-timed and sequential respiratory events orchestrated by complex neuromuscular networking of the diaphragm, chest and abdominal respiratory muscles, the glottis and parts of the brain.


In Operando Study of the Hydrogen-Induced Switching of Magnetic Anisotropy at the Co/Pd Interface for Magnetic Hydrogen Gas Sensing.
Heterostructures exhibiting perpendicular magnetic anisotropy (PMA) have traditionally served the magnetic recording industry. However, an opportunity exists to expand the applications of PMA heterostructures into the realm of hydrogen sensing using ferromagnetic resonance (FMR) by exploiting the hydrogen-induced modifications to PMA that occur at the interface between Pd and a ferromagnet. Here, we present the first in operando depth-resolved study of the in-plane interfacial magnetization of a Co/Pd film which features tailorable PMA in the presence of hydrogen gas. We combine polarized neutron reflectometry with in situ FMR to explore how the absorption of hydrogen at the Co/Pd interface affects the heterostructures spin-resonance condition during hydrogen cycling. Experimental data and modeling reveal that the Pd layer expands when exposed to hydrogen gas, while the in-plane magnetic moment of the Co/Pd film increases as the interfacial PMA is reduced to affect the FMR frequency. This work highlights a potential route for magnetic hydrogen gas sensing.


Differences and similarities between gay and straight individuals involved in the sadomasochistic subculture.
Results from some new analyses as well as a selective review of the results of six empirical studies on a self-identified sample of sadomasochistically-oriented individuals (22 women and 162 men) with an emphasis on differences between gay and straight participants are presented. The gay male respondents seemed to be better educated, to hold white-collar occupations and to be more sadistically oriented. The gay male respondents became aware of their sadomasochistic preferences and had their first experiences at an older age. They showed a preference for leather outfits, anal intercourse, rimming, dildos, wrestling, special equipment and uniform scenes, and the straight male respondents for verbal humiliation, mask and blindfold, gags, rubber outfits, cane whipping, vaginal intercourse, cross dressing, and straitjackets, and different role plays except for uniform scenes. Four separate sexual themes were identified: hypermasculinity; administration and receiving of pain; physical restriction and psychological humiliation. Gay male participants showed a preference for hypermasculinity and straight men respondents for humiliation. The number of lesbian respondents was too low to draw any definite conclusions. Differences between gay and straight male sadomasochists in the present sample were found. Clearly, sadomasochism cannot be thought of as a unitary phenomenon: People who identify themselves as sadomasochists mean different things by these identifications.


Towards a national strategy for the provision of spiritual care during major health disasters: A qualitative study.
Spirituality is beneficial to health. Evidence around the benefits of Spiritual care (SC) is advancing, and training is becoming part of healthcare professional development. As the COVID-19 crisis showed, during major health disasters (MHDs), the demand for SC grows exponentially, while the burden of care and focus on preserving life often hamper its provision. Nonetheless, existing health emergency strategic frameworks lack preparedness for the provision of SC. The aim of this study was to identify the components for a National Strategy (NS) for the provision of SC during MHDs. Descriptive, cross-sectional, qualitative phenomenological design based on individual, semi-structured e-interviews with nursing managers and National Health Service/volunteer chaplains based in England. Thematic analysis of 25 e-interview data was performed based on a dialogic collaborative process. Eleven themes were identified as components of the proposed NS. From these components, specific recommendations for practical actions are provided. An integrated framework approach and smart investments in resources, staff training and technologies should be led by the paradigm of culturally competent and compassionate care. The need to have strategic frameworks, both national and local, that better equip a country healthcare sector to prevent, face, and recover from MHDs is paramount. Catering for the spiritual needs of the affected population should be a key aspect of any health emergency strategy to ensure the preservation of quality care.


Procalcitonin can be used as a marker of premature atherosclerosis in acromegaly.
The objective of the study was to evaluate arterial morphologic changes of early atherosclerosis and changes in procalcitonin (PCT) levels in patients with acromegaly according to disease activity. Thirty-three active and 20 inactive acromegaly patients followed at Endocrinology-Metabolism out-patient clinic of Cerrahpasa Medical Faculty between 2004 and 2008 were included in the study. Twenty gender and age matched healthy subjects were included as the control group. Intima-media thickness (IMT) of the carotid arteries was measured by ultrasonography. Blood was drawn for biochemical tests and the serum concentrations of C-reactive protein (CRP) and PCT. Intergroup analysis revealed no significant differences between Growth hormone (GH), insulin like growth factor-1 (IGF-1), and IMT (P = 0.42, P = 0.47 respectively). No significant differences were found in the fibrinogen, CRP and PCT levels of the acromegaly patients and the subjects in the control group (P = 0.57, P = 0.84, P = 0.68 respectively). In the patients with IMT ≥ 1 mm, PCT (0.4 [IQR: 0.4-0.55]) levels were significantly different from the patients without atherosclerosis (0.06 [IQR: 0.05-0.12], P < 0.001). The correlation between IMT and PCT (P = 0.001, r = 0.47) was more significant than the correlation between IMT and CRP (P = 0.01, r = 0.28). There was a positive correlation between IMT and atherosclerotic risk factors such as age (P = 0.01, r = 0.27) and body mass index (BMI; P = 0.005, r = 0.32). Our results showed that PCT increases before CRP and it can be useful for the assessment of premature atherosclerosis in acromegaly as well.


Recurrence of prostate cancer after HIFU. Proposal of a novel predictive index.
Prostate cancer is one of the most common cancers in men over 50 years of age. Surgery, radiotherapy and hormonal manipulation represent its typical treatment. High-Intensity Focused Ultrasound (HIFU) is an alternative choice in localized prostate cancer. To date, an index for prediction of recurrence in patients treated with HIFU is not availabe. Our study proposes a novel index for the predition of recurrence able to determine if a candidate is fit for this tratment. 107 patients underwent HIFU fram 2010 to 2015. A total of 12 variables were considered for the analysis. The final predictive model was obtained through a stepwise forward selection method. The final model used a total of 6 variables, all correlated to the response variable. The Index is able to predict the recurrence after HIFU tratment in the most majority of candidates to treatment. The index may be used to make a more scientific decision with regard to choosing optimal candidates for HIFU.


A portable system for identifying urinary tract infection in primary care using a PC-based chromatic technique.
An approach is described for monitoring urine samples using a portable system based on chromatic techniques and for predicting urinary tract infection (UTI) from the results. The system uses a webcam-computer combination with the screen of a computer visual display unit as a tuneable illumination source. It is shown that the system can operate in a robust manner under ambient lighting conditions and with potential for use as a point of care test in primary care. The present approach combines information on urine liquid concentration and turbidity. Its performance in an exploratory study is compared with microbiological culture of 200 urine samples, of which 79 had bacterial growth >10(5) colony forming unit/millilitre (cfu ml(-1)) indicative of UTI. It is shown that both sensitivity and negative predictive value of 0.92 could be achieved.


The Sam68 nuclear body is composed of two RNase-sensitive substructures joined by the adaptor HNRNPL.
The mammalian cell nucleus contains membraneless suborganelles referred to as nuclear bodies (NBs). Some NBs are formed with an architectural RNA (arcRNA) as the structural core. Here, we searched for new NBs that are built on unidentified arcRNAs by screening for ribonuclease (RNase)-sensitive NBs using 32,651 fluorescently tagged human cDNA clones. We identified 32 tagged proteins that required RNA for their localization in distinct nuclear foci. Among them, seven RNA-binding proteins commonly localized in the Sam68 nuclear body (SNB), which was disrupted by RNase treatment. Knockdown of each SNB protein revealed that SNBs are composed of two distinct RNase-sensitive substructures. One substructure is present as a distinct NB, termed the DBC1 body, in certain conditions, and the more dynamic substructure including Sam68 joins to form the intact SNB. HNRNPL acts as the adaptor to combine the two substructures and form the intact SNB through the interaction of two sets of RNA recognition motifs with the putative arcRNAs in the respective substructures.


Interphase molecular cytogenetic screening for chromosomal abnormalities of prognostic significance in childhood acute lymphoblastic leukaemia: a UK Cancer Cytogenetics Group Study.
Summary Interphase fluorescence in situ hybridization (iFISH) was used independently to reveal chromosomal abnormalities of prognostic importance in a large, consecutive series of children (n = 2367) with acute lymphoblastic leukaemia (ALL). The fusions, TEL/AML1 and BCR/ABL, and rearrangements of the MLL gene occurred at frequencies of 22% (n = 447/2027) (25% in B-lineage ALL), 2% (n = 43/2027) and 2% (n = 47/2016) respectively. There was considerable variation in iFISH signal patterns both between and within patient samples. The TEL/AML1 probe showed the highest incidence of variation (59%, n = 524/884), which included 38 (2%) patients with clustered, multiple copies of AML1. We were thus able to define amplification of AML1 as a new recurrent abnormality in ALL, associated with a poor prognosis. Amplification involving the ABL gene, a rare recurrent abnormality confined to T ALL patients, was identified for the first time. The use of centromeric probes revealed significant hidden high hyperdiploidy of 33% and 59%, respectively, in patients with normal (n = 21/64) or failed (n = 32/54) cytogenetic results. The iFISH contributed significantly to the high success rate of 91% (n = 2114/2323) and the remarkable abnormality detection rate of 89% (n = 1879/2114). This study highlights the importance of iFISH as a complementary tool to cytogenetics in routine screening for significant chromosomal abnormalities in ALL.


The use of a dipstick antigen-capture assay for the diagnosis of Plasmodium falciparum infection in a remote forested area of central India.
A study was carried out in a highly malarious forested belt of central India (Madhya Pradesh) to evaluate the usefulness of a dipstick antigen-capture assay (ParaSight F) as a diagnostic method for Plasmodium falciparum appropriate for field use. In all, 1,231 patients with fever were screened in parallel with the taking of thick blood smears. The sensitivity and specificity of the dipstick test for detection of P. falciparum were 93% and 92.5%, respectively. Results also indicate that the dipstick test result became negative within seven days after initiation of curative chemotherapy in the majority of cases (95%). Tribal populations in remote forested areas are important reservoirs of P. falciparum that are mostly resistant to standard antimalarial drugs and a source of malaria to the rest of the country. Control of malaria is limited by inaccessibility, lack of medical services, public ignorance of the disease, and other technical problems. In this population, the dipstick assay was found to be highly accurate, simple, and rapid, suggesting that it may be used in remote forested areas without the need for microscopic examination. The simplicity of the dipstick test enabled administration of more effective and expensive antimalarials preventing further buildup and dissemination of resistant parasites and a return of symptoms.


Validation of Blood Volume Fraction Quantification with 3D Gradient Echo Dynamic Contrast-Enhanced Magnetic Resonance Imaging in Porcine Skeletal Muscle.
The purpose of this study was to assess the accuracy of fractional blood volume (vb) estimates in low-perfused and low-vascularized tissue using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI). The results of different MRI methods were compared with histology to evaluate the accuracy of these methods under clinical conditions. vb was estimated by DCE-MRI using a 3D gradient echo sequence with k-space undersampling in five muscle groups in the hind leg of 9 female pigs. Two gadolinium-based contrast agents (CA) were used: a rapidly extravasating, extracellular, gadolinium-based, low-molecular-weight contrast agent (LMCA, gadoterate meglumine) and an extracellular, gadolinium-based, albumin-binding, slowly extravasating blood pool contrast agent (BPCA, gadofosveset trisodium). LMCA data were evaluated using the extended Tofts model (ETM) and the two-compartment exchange model (2CXM). The images acquired with administration of the BPCA were used to evaluate the accuracy of vb estimation with a bolus deconvolution technique (BD) and a method we call equilibrium MRI (EqMRI). The latter calculates the ratio of the magnitude of the relaxation rate change in the tissue curve at an approximate equilibrium state to the height of the same area of the arterial input function (AIF). Immunohistochemical staining with isolectin was used to label endothelium. A light microscope was used to estimate the fractional vascular area by relating the vascular region to the total tissue region (immunohistochemical vessel staining, IHVS). In addition, the percentage fraction of vascular volume was determined by multiplying the microvascular density (MVD) with the average estimated capillary lumen, [Formula: see text], where d = 8μm is the assumed capillary diameter (microvascular density estimation, MVDE). Except for ETM values, highly significant correlations were found between most of the MRI methods investigated. In the cranial thigh, for example, the vb medians (interquartile range, IQRs) of IHVS, MVDE, BD, EqMRI, 2CXM and ETM were vb = 0.7(0.3)%, 1.1(0.4)%, 1.1(0.4)%, 1.4(0.3)%, 1.2(1.8)% and 0.1(0.2)%, respectively. Variances, expressed by the difference between third and first quartiles (IQR) were highest for the 2CXM for all muscle groups. High correlations between the values in four muscle groups-medial, cranial, lateral thigh and lower leg - estimated with MRI and histology were found between BD and EqMRI, MVDE and 2CXM and IHVS and ETM. Except for the ETM, no significant differences between the vb medians of all MRI methods were revealed with the Wilcoxon rank sum test. The same holds for all muscle regions using the 2CXM and MVDE. Except for cranial thigh muscle, no significant difference was found between EqMRI and MVDE. And except for the cranial thigh and the lower leg muscle, there was also no significant difference between the vb medians of BD and MVDE. Overall, there was good vb agreement between histology and the BPCA MRI methods and the 2CXM LMCA approach with the exception of the ETM method. Although LMCA models have the advantage of providing excellent curve fits and can in principle determine more physiological parameters than BPCA methods, they yield more inaccurate results.


Potential biomarkers of fatigue identified by plasma metabolome analysis in rats.
In the present study, prior to the establishment of a method for the clinical diagnosis of chronic fatigue in humans, we validated the utility of plasma metabolomic analysis in a rat model of fatigue using capillary electrophoresis-mass spectrometry (CE-MS). In order to obtain a fatigued animal group, rats were placed in a cage filled with water to a height of 2.2 cm for 5 days. A food-restricted group, in which rats were limited to 10 g/d of food (around 50% of the control group), was also assessed. The food-restricted group exhibited weight reduction similar to that of the fatigued group. CE-MS measurements were performed to evaluate the profile of food intake-dependent metabolic changes, as well as the profile in fatigue loading, resulting in the identification of 48 metabolites in plasma. Multivariate analyses using hierarchical clustering and principal component analysis revealed that the plasma metabolome in the fatigued group showed clear differences from those in the control and food-restricted groups. In the fatigued group, we found distinctive changes in metabolites related to branched-chain amino acid metabolism, urea cycle, and proline metabolism. Specifically, the fatigued group exhibited significant increases in valine, leucine, isoleucine, and 2-oxoisopentanoate, and significant decreases in citrulline and hydroxyproline compared with the control and food-restricted groups. Plasma levels of total nitric oxide were increased in the fatigued group, indicating systemic oxidative stress. Further, plasma metabolites involved in the citrate cycle, such as cis-aconitate and isocitrate, were reduced in the fatigued group. The levels of ATP were significantly decreased in the liver and skeletal muscle, indicative of a deterioration in energy metabolism in these organs. Thus, this comprehensive metabolic analysis furthered our understanding of the pathophysiology of fatigue, and identified potential diagnostic biomarkers based on fatigue pathophysiology.