Urinary Peptidomic Biomarker for Personalized Prevention and Treatment of Diastolic Left Ventricular Dysfunction.
Diastolic heart failure (DHF) is characterized by slow left ventricular (LV) relaxation, increased LV stiffness, interstitial deposition of collagen, and a modified extracellular matrix proteins. Among Europeans, the frequency of asymptomatic diastolic LV dysfunction (DD) is 25%. This constitutes a large pool of people at high risk of DHF. The goal of this review was to describe the discovery and the initial validation of new multidimensional urinary peptidomic biomarkers (UPB) indicative of DD, mainly consisting of collagen fragments, and to describe a roadmap for their introduction into clinical practice. The availability of new drugs creates a window of opportunity for mounting a randomized clinical trial consolidating the clinical applicability of UPB to screen for DD. If successfully completed, such trial will benefit ≈25% of all people older than 50 years and open a large market for a UPB diagnostic tool and the drug tested. Moreover, sequenced peptides making up UPB will generate novel insights in the pathophysiology of DD and facilitate personalized treatment of patients with DHF for whom prevention came too late. If proven cost-effective, the clinical application of UPB will contribute to the sustainability of health care in aging population in epidemiologic transition.


Morbidly obese woman unaware of pregnancy until full-term and complicated by intraamniotic sepsis with pseudomonas.
A 32-year-old Caucasian woman of body mass index (BMI) 46 presented with urinary symptoms to accident and emergency (A&E). Acute pyelonephritis was the diagnosis. Transabdominal scan revealed a live term fetus. Both the partners were unaware of the ongoing pregnancy until diagnosed. She underwent emergency cesarean under general anaesthesia (GA) for nonreassuring CTG, severe chorioamnionitis, and moderate preecclampsia. A live male baby weighing 4400 grams delivered in poor condition. Placental tissue on culture exhibited scanty growth of pseudomonas aeruginosa. Chorioamnionitis due to pseudomonas is rare, with high neonatal morbidity and mortality. It is mostly reported among preterm prelabor rupture of membranes (PPROM). Educating the community especially morbidly obese women if they put on excessive weight or with irregular periods should seek doctor's advice and exclude pregnancy. For the primary care provider, it is of great importance to exclude pregnancy in any reproductive woman presenting with abdominal complaints. This case also brings to clinicians notice that pseudomonas can be community-acquired and can affect term pregnancies with intact or prolonged rupture of membranes.


Body mapping of sweating patterns in athletes: a sex comparison.
Limited regional sweat rate (RSR) data are available for females, with only a small number of sites measured across the body. Similarly, sex differences in sweating concentrate on whole body values, with limited RSR data available. A modified absorbent technique was used to collect sweat at two exercise intensities (60% (I1) and 75% (I2) V˙O2max) in 13 aerobically trained females (21 ± 1 yr, 59.5 ± 10 mL·min·kg V˙O2max) in moderately warm conditions (25°C, 45% relative humidity, 2 m·s air velocity). Females were compared with nine aerobically trained males (23 ± 3 yr, 70.2 ± 13 mL·min·kg V˙O2max) tested under the same conditions. Female I1 RSR was highest at the central upper back, heels, and dorsal foot and between the breasts (223, 161, 139, and 139 g·m·h, respectively). Lowest values were over the breasts and the middle and lower outer back (<16 g·m·h). At I2, the central upper back, bra triangle, and lower back showed the highest RSR (723, 470, and 333 g·m·h, respectively). Regions of the breasts and palms had the lowest RSR at I2 (<82 g·m·h). Significantly greater gross sweat loss and thus RSR were observed in males versus females at both exercise intensities. For the same metabolic heat production (male I1 vs female I2), absolute and normalized RSR showed a significant region-sex interaction (P < 0.001), with a greater distribution toward the arms and hands in females versus males. Despite some differences in distribution, both sexes showed highest RSR on the central upper back and the lowest toward the extremities. No correlation was observed between local skin temperature and RSR, failing to explain RSR variation observed. These data have important applications for sex-specific clothing design, thermophysiological modeling, and thermal manikin design.


Conjugation of Palbociclib with MHI-148 Has an Increased Cytotoxic Effect for Breast Cancer Cells and an Altered Mechanism of Action.
The CDK4/6 inhibitor palbociclib, combined with endocrine therapy, has been shown to be effective in postmenopausal women with estrogen receptor-positive, HER2-negative advanced or metastatic breast cancer. However, palbociclib is not as effective in the highly aggressive, triple-negative breast cancer that lacks sensitivity to chemotherapy or endocrine therapy. We hypothesized that conjugation of the near-infrared dye MHI-148 with palbociclib can produce a potential theranostic in triple-negative, as well as estrogen receptor-positive, breast cancer cells. In our study, the conjugate was found to have enhanced activity in all mammalian cell lines tested in vitro. However, the conjugate was cytotoxic and did not induce G1 cell cycle arrest in breast cancer cells, suggesting its mechanism of action differs from the parent compound palbociclib. The study highlights the importance of investigating the mechanism of conjugates of near-infrared dyes to therapeutic compounds, as conjugation can potentially result in a change of mechanism or target, with an enhanced cytotoxic effect in this case.


Acute effects of lipopolysaccharide (LPS) in kidney of rats and preventive role of vitamin E and sodium selenite.
Lipopolysaccharide (LPS) as an endotoxin forms part of the cell wall of gram-negative bacteria and is responsible for initiating an acute inflammation after entering the living tissue. In this study, male rats were divided into eight groups: control group, vitamin E (VE) treatment group (200 mg/kg body weight (b.w.)), sodium selenite (SS) treatment (0.35 mg/kg b.w.) group, VE + SS treatment group (200 + 0.35 mg/kg b.w.), LPS treatment group (10 mg/kg b.w.), LPS + VE (10+200 mg/kg b.w.), LPS + SS treatment (10 + 0.35 mg/kg b.w.), and LPS + SS + VE treatment (10 + 0.35 + 200 mg/kg b.w.) group. Oxidative stress parameters, pathological changes, immunohistochemical analyses, terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end-labeling (TUNEL) assay, and changes in DNA structure with comet assay of the kidney were investigated at the end 6 h comparatively with the control group. When LPS-treated group was compared with the control group, antioxidant enzyme activities were decreased and malondialdehyde (MDA) levels, changes in histological and DNA structure and apoptosis were increased significantly at the end of 6 h. However, when LPS + SS and/or VE-treated group were compared with the LPS-treated group, superoxide dismutase, catalase, glutathione peroxidase, and glutathione- S-transferase activities were increased and MDA levels were decreased significantly at the end of the treatment period. Light investigations figured out pathological changes in kidneys of LPS- and LPS + SS and/or VE-treated groups. There was a decrease in the number of proliferating cell nuclear antigen-positive cells and an increase in the number of TUNEL-positive apoptotic cells in the wall of the distal and proximal tubules. As a result, it was observed that the combined use of antioxidants was more protective than their use alone against LPS.


TGF-β-Induced TMEPAI Attenuates the Response of Triple-Negative Breast Cancer Cells to Doxorubicin and Paclitaxel.
Triple-negative breast cancer (TNBC) is a refractory type of breast cancer with poor prognosis and limited choice for treatment. Previous studies had shown that TNBC has high expressions of transmembrane prostate androgen-induced protein (TMEPAI). TMEPAI was known to be induced by TGF-β/Smad signaling and have tumorigenic functions that converting TGF-β from tumor suppressor to tumor promoter and inducing epithelial-mesenchymal transition (EMT). Therefore, we aimed to define the role of TMEPAI in triple-negative breast cancer cells treatment using several anti-cancers in the presence of TGF-β. TMEPAI-knock out (KO) was carried out in a triple-negative breast cancer cell, BT549. TMEPAI editing was developed using the CRISPR-Cas9 system using two combinations of sgRNA to remove exon 4 of the TMEPAI gene entirely. Genotyping and proteomic analysis were performed to check the establishment of the TMEPAI-KO cells. Wild type (WT) and KO cells were used to determine inhibitory concentration 50% (IC50) of several anti-cancers: doxorubicin, cisplatin, paclitaxel, and bicalutamide in the presence of TGF-β treatment. KO cells were successfully established by completely removing the TMEPAI gene, which was proven in genomic and proteomic analysis. Further, in TMEPAI-KO cells, we found a significant reduction of IC50 for doxorubicin and paclitaxel, and minimal effects were seen for cisplatin and bicalutamide. Our findings suggest that TGF-β-induced TMEPAI attenuates the response of TNBC to doxorubicin and paclitaxel, but not to cisplatin and bicalutamide. TGF-β induced TMEPAI contributes to the reduced response of TNBC treatment to doxorubicin and paclitaxel, but minimal on cisplatin and bicalutamide. Further study is needed to confirm our findings in other growth factor-induced cells, as well as in in vivo model.


[Efficacies of fluid resuscitation as guided by lactate clearance rate and central venous oxygen saturation in patients with septic shock].
To estimate the efficacies of fluid resuscitations as guided by lactate clearance rate (LC) and central venous oxygen saturation (ScvO₂) in patients with septic shock. 100 patients diagnosed with septic shock from June 2012 to June 2014 in department of critical care medicine of sixteen hospitals were enrolled. They were randomly divided into two groups of study and control (each n = 50). After a diagnosis of sepsis shock, they were treated symptomatically timely and fluid resuscitation was started as early as possible according to the 2008 Guideline for Managing Sepsis & Septic Shock. Central venous pressure (CVP) ≥ 8 mmHg (1 mmHg = 0.133 kPa), mean arterial pressure (MAP) ≥ 65 mmHg, urine output ≥ 0.5 ml × kg⁻¹ × h⁻¹, ScvO₂≥ 70% and LC ≥ 10% (or lactate ≤ 2.0 mmol) served as target values for fluid resuscitation therapy in study group versus CVP ≥ 8 mmHg, MAP ≥ 65 mmHg, urine output ≥ 0.5 ml × kg⁻¹ × h⁻¹ and ScvO₂≥ 70% in control group. The general conditions and clinical characteristics, changes in CVP, MAP, urine output, ScvO₂, lactate level and/or LC before (0 hour) and every hour (1-6 h) after the start of fluid resuscitation and other related outcome indicators were recorded. No significant difference existed in general data. The 28-day mortality was 40% for study group versus 56% for control group. There was no significant inter-group difference (P > 0.05). The time of mechanical ventilation and length of intensive care unit (ICU) stay were lower in study group than those in control group [mechanical ventilation time (11.200 ± 17.069) vs (15.760 ± 14.215), P = 0.150; length of ICU stay (13.240 ± 17.127) vs (23.980 ± 18.298), P = 0.003]. The 28-day mortality was independently associated with LC and ScvO₂reaching target values for fluid resuscitation in study group (χ² = 10.930, P = 0.001) while the 28-day mortality was independently associated with ScvO₂reaching target value for fluid resuscitation in control group (χ² = 6.395, P = 0.011). Among all patients, the 28-day mortality was independently associated with ScvO₂reaching target value for fluid resuscitation (χ² = 14.530, P = 0.000), but not LC (χ² = 1.175, P = 0.278). A combination of LC and ScvO₂may serve an index in confirming the endpoint of fluid resuscitation for patients with septic shock. Fluid resuscitation therapy under the guidance of LC and ScvO₂is more accurate and reliable than the guidance of ScvO₂alone.


Alpha thalassaemia in tribal communities of coastal Maharashtra, India.
In a routine community health survey conducted in adult Adivasis of the costal Maharashtra, microcytosis and hyprochromia were observed in more than 80 per cent of both males and females having normal haemoglobin levels suggesting the possibility of α-thalassaemia in these communities. We conducted a study in Adivasi students in the same region to find out the magnitude of α-thalessaemia. The participants (28 girls and 23 boys) were 14-17 yr old studying in a tribal school. Fasting venous blood samples (5 ml) were subjected to complete blood count (CBC), Hb-HPLC and DNA analysis using gap-PCR for deletion of -α3.7 and -α4.2, the two most common molecular lesions observed in α-thalassaemia in India. Microcytic hypochromic anaemia was observed 50 and 35 per cent girls and boys, respectively. Iron supplementation improved Hb levels but did not correct microcytois and hypochromia. m0 ore than 80 per cent non-anaemic students of both sexes showed microcytois and hypochromia. DNA analysis confirmed that the haematological alterations were due to α-thalassaemia trait characterized by deletion of -α3.7. Majority (>60%) of the affected students had two deletions (-α3.7/-α3.7 genotype α+ thalassaemia. This is perhaps the first report on the occurrence of α-thalassaemia in tribal communities of coastal Maharashtra. Very high (78.4%) haplotype frequency of -α3.7 suggests that the condition is almost genetically fixed. These preliminary observations should stimulate well planned large scale epidemiological studies on α-thalassaemia in the region.


Who takes paternity leave? A cohort study on prior social and health characteristics among fathers in Stockholm.
Progress towards gender equality involves changes in the traditional parental division - female caring and male breadwinning. One aspect is increased parental leave for fathers, which may benefit the health of mothers, children, and fathers themselves. We examined how social and health characteristics (2002) were associated with paternity leave in excess of the 'father quota' of 60 days (2003-2006) in the Stockholm Public Health Cohort. Generally, fathers with stable social position, fit lifestyles, and good health had increased chances of paternity leave uptake. Our findings may contribute to identifying target groups for parental leave strategies among fathers; they indicate also that research on gender equality and public health must carefully address the problems of confounding and health-related selection.


Effects of bariatric surgery on cardiorespiratory fitness: A systematic review and meta-analysis.
Although bariatric surgery (BS) is recognized as an effective strategy for body weight loss, its impact on cardiorespiratory fitness (CRF) is still unclear. We aimed to examine postoperative changes in CRF (VO2max/peak ) and its relationship with weight loss among adults undergoing BS. We systematically searched the WoS, PubMed, MEDLINE, and Scopus databases. Observational and intervention studies were selected reporting the presurgery and postsurgery CRF, measured by breath-by-breath VO2 or its estimation. Eleven articles (312 patients) revealed that BS leads to a reduction in absolute VO2max/peak in the short term (effect size, ES = -0.539; 95%CI = -0.708, -0.369; p < 0.001), and those patients who suffered a more significant decrease in BMI after BS also had a greater loss of absolute VO2max/peak . However, VO2max/peak relative to body weight increased after surgery (ES = 0.658; 95%CI = 0.473, 0.842; p < 0.001). An insufficient number of studies were found investigating medium and long-term changes in CRF after BS. This study provides moderate-quality evidence that the weight loss induced by BS can reduce CRF in the short term, which represents a therapeutic target to optimize BS outcomes. More high-quality studies are needed to evaluate the impact of BS on VO2max/peak in the short, medium, and long term including normalized values for fat-free mass.


The multidimensional physiological responses to postconditioning.
Reperfusion is the definitive treatment to reduce infarct size and other manifestations of postischemic injury. However, reperfusion contributes to postischemic injury, and, therefore, reperfusion therapies do not achieve the optimal salvage of myocardium. Other tissues as well undergo injury after reperfusion, notably, the coronary vascular endothelium. Postconditioning has been shown to have salubrious effects on different tissue types within the heart (cardiomyocytes, endothelium) and to protect against various pathologic processes, including necrosis, apoptosis, contractile dysfunction, arrhythmias, and microvascular injury or "no-reflow." The mechanisms by which postconditioning alters the pathophysiology of reperfusion injury is exceedingly complex and involves physiological mechanisms (e.g., delaying re-alkalinization of tissue pH, triggering release of autacoids, and opening and closing of various channels) and molecular mechanisms (activation of kinases) that affect cellular and subcellular targets or effectors. The physiologic responses to postconditioning are not isolated or mutually exclusive, but are interactive, with one response affecting another in an integrated manner. This integrated response on multiple targets differs from the monotherapy approach by drugs that have failed to reduce reperfusion injury on a consistent basis and may underlie the efficacy of this therapeutic approach across species and in human trials.


Low Recombination Proficiency Score (RPS) Predicts Heightened Sensitivity to DNA-Damaging Chemotherapy in Breast Cancer.
Purpose: Molecular-based cancer tests have been developed to augment the standard clinical and pathologic features used to tailor treatments to individual breast cancer patients. Homologous recombination (HR) repairs double-stranded DNA breaks and promotes tolerance to lesions that disrupt DNA replication. Recombination Proficiency Score (RPS) quantifies HR efficiency based on the expression of four genes involved in DNA damage repair. We hypothesized low RPS values can identify HR-deficient breast cancers most sensitive to DNA-damaging chemotherapy.Experimental Design: We collected pathologic tumor responses and tumor gene expression values for breast cancer patients that were prospectively enrolled on clinical trials involving preoperative chemotherapy followed by surgery (N = 513). We developed an algorithm to calculate breast cancer-specific RPS (RPSb) values on an individual sample basis.Results: Low RPSb tumors are approximately twice as likely to exhibit a complete pathologic response or minimal residual disease to preoperative anthracycline-based chemotherapy as compared with high RPSb tumors. Basal, HER2-enriched, and luminal B breast cancer subtypes exhibit low RPSb values. In addition, RPSb predicts treatment responsiveness after controlling for clinical and pathologic features, as well as intrinsic breast subtype.Conclusions: Overall, our findings indicate that low RPS breast cancers exhibit aggressive features at baseline, but they have heightened sensitivity to DNA-damaging chemotherapy. Low RPSb values in basal, HER2-enriched, and luminal B subtypes provide a mechanistic explanation for their clinical behaviors and genomic instability. RPSb augments standard clinical and pathologic features used to tailor treatments, thereby enabling more personalized treatment strategies for individual breast cancer patients. Clin Cancer Res; 23(15); 4493-500. ©2017 AACR.


Ultra-thin Ga nanosheets: analogues of high pressure Ga(iii).
Ultra-thin Ga islands of β-Ga(110), high-pressure phase Ga(iii) and a new phase of stripe superstructure are obtained on Si(111). STM combined with theoretical calculations suggests that the stripe superstructure originates from Ga(iii) (001) with stacking rearrangement. This work provides a new strategy for synthesizing low-dimensional nanomaterials and accessing high pressure phases.


Estimation of muscle response using three-dimensional musculoskeletal models before impact situation: a simulation study.
When car crash experiments are performed using cadavers or dummies, the active muscles' reaction on crash situations cannot be observed. The aim of this study is to estimate muscles' response of the major muscle groups using three-dimensional musculoskeletal model by dynamic simulations of low-speed sled-impact. The three-dimensional musculoskeletal models of eight subjects were developed, including 241 degrees of freedom and 86 muscles. The muscle parameters considering limb lengths and the force-generating properties of the muscles were redefined by optimization to fit for each subject. Kinematic data and external forces measured by motion tracking system and dynamometer were then input as boundary conditions. Through a least-squares optimization algorithm, active muscles' responses were calculated during inverse dynamic analysis tracking the motion of each subject. Electromyography for major muscles at elbow, knee, and ankle joints was measured to validate each model. For low-speed sled-impact crash, experiment and simulation with optimized and unoptimized muscle parameters were performed at 9.4 m/h and 10 m/h and muscle activities were compared among them. The muscle activities with optimized parameters were closer to experimental measurements than the results without optimization. In addition, the extensor muscle activities at knee, ankle, and elbow joint were found considerably at impact time, unlike previous studies using cadaver or dummies. This study demonstrated the need to optimize the muscle parameters to predict impact situation correctly in computational studies using musculoskeletal models. And to improve accuracy of analysis for car crash injury using humanlike dummies, muscle reflex function, major extensor muscles' response at elbow, knee, and ankle joints, should be considered.


Electronic response of a (P,N)-based ligand on metal coordination.
The reaction of [(THF)Li(Ph(2)PC(H)Py)] with ZnCl(2) in the presence of ZnO yields the zinc complex [Zn(3)(Ph(2)PC(H)Py)(4)O] (1). Deprotonation of the phosphane Ph(2)P(CH(2)Py) with [Fe(N(SiMe(3))(2))2] gives the iron complexes [(Ph(2)P(CH(2)Py))Fe(Ph(2)PC(H)Py)2] (2) and [Fe(Ph(2)PC(H)Py)(N(SiMe(3))(2))]2 (3), depending on the ratio of phosphane. The solid state structures of the metal complexes illustrate the coordination flexibility of the [Ph(2)C(H)Py](-)-anion. Depending on the electronic requirements of the coordinated metal the anion acts as a (P,N)-chelating amide or C-coordinating carbanion with the P- and N-heteroatoms as donor bases.


Topological Control of Magnetic Textures.
A micromagnetic study is carried out on the role of using topology to stabilize different magnetic textures, such as a vortex or an anti-vortex state, in a magnetic heterostructure consisting of a Permalloy disk coupled to a set of nanomagnetic bars. The topological boundary condition is set by the stray field contributions of the nanomagnet bars and thus by their magnetization configuration, and can be described by a discretized winding number that will be matched by the winding number of the topological state set in the disk. The lowest number of nanomagnets that defines a suitable boundary is four, and we identify a critical internanomagnet angle of 225° between two nanomagnets, at which the boundary fails because the winding number of the nanomagnet configuration no longer controls that of the disk magnetization. The boundary also fails when the disk-nanomagnets separation is > 50 nm and for disk diameters > 480 nm. Finally, we provide preliminary experimental evidence from magnetic force microscopy studies in which we demonstrate that an energetically unstable, anti-vortex-like structure can indeed be stabilized in a Permalloy disk, provided that the appropriate topological conditions are set.


Childhood homicide, suicide, and firearm deaths: an international comparison.
This article is intended to provide a better understanding of the incidence of violent deaths among children under 15 years of age in highly industrialized countries/areas. We found that rates of violent childhood deaths are not uniform in the industrialized world and that rates in the United States greatly exceed those in the other countries and areas. In fact, total firearm deaths among children are 12 times higher in the United States than in all of the other countries or areas combined; childhood homicide rates are 5 times higher; and childhood suicide rates are twice as high. Five countries or areas, 3 of which are in Asia, reported no firearm deaths among children under 15 years old. These findings suggest the value of conducting further research to explore these cross-national differences.


Leukotrienes and tyrosine phosphorylation mediate stretching-induced actin cytoskeletal remodeling in endothelial cells.
We studied actin cytoskeletal remodeling and the role of leukotrienes and tyrosine phosphorylation in the response of endothelial cells to different types of cyclic mechanical stretching. Human aortic endothelial cells were grown on deformable silicone membranes subjected to either cyclic one-directional (strip) stretching (10%, 0.5 Hz), or biaxial stretching. After 1 min of either type of stretching, actin cytoskeletons of the stretched cells were already disrupted. After stretching for 10 and 30 min, the percentage of the stretched cells that had disrupted actin cytoskeletons were significantly increased, compared with control cells without stretching. Also, at these two time points, biaxial stretching consistently produced higher frequencies of actin cytoskeleton disruption. At 3 h, strip stretching caused the formation of stress fiber bundles, which were oriented nearly perpendicular to the stretching direction. With biaxial stretching, however, actin cytoskeletons in many stretched cells were remodeled into three-dimensional actin structures protruding outside the substrate plane, within which cyclic stretching was applied. In both stretching conditions, actin filaments were formed in the direction without substrate deformation. Moreover, substantially inhibiting either leukotriene production with nordihydroguaiaretic acid or tyrosine phosphorylation with tyrphostin A25 did not block the actin cytoskeletal remodeling. However, inhibiting both leukotriene production and tyrosine phosphorylation completely blocked the actin cytoskeletal remodeling. Thus, the study showed that the remodeling of actin cytoskeletons of the stretched endothelial cells include rapid disruption first and then re-formation. The resulting pattern of the actin cytoskeleton after remodeling depends on the type of cyclic stretching applied, but under either type of cyclic stretching, the actin filaments are formed in the direction without substrate deformation. Finally, leukotrienes and tyrosine phosphorylation are necessary for actin cytoskeletal remodeling of the endothelial cells in response to mechanical stretching.


Adding specialized clinics for remote-dwellers with chronic kidney disease: a cost-utility analysis.
This study aimed to determine whether opening a new clinic in a remote region would be a cost-effective means of improving care for remote-dwellers with CKD. This study is a cost-utility analysis from a public payer's perspective over a lifetime horizon, using administrative data from a large cohort of adults with stage 3b-4 CKD in Alberta, Canada. The association between the distance from each simulated patient's residence and the practice location of the closest nephrologist and clinical outcomes (quality of care, hospitalization, dialysis, and death) were examined. A Markov 6-month cycle economic decision model was analyzed; estimates of the effect of a new clinic were based on the association between residence location, resource use, and outcomes. Costs are reported in 2009 Canadian dollars. The costs for equipping and operating a clinic for 321 remote-dwelling patients were estimated at $25,000 and $250,000/yr, respectively. The incremental cost-utility ratios (ICURs) ranged from $4000 to $8000/quality-adjusted life-year under most scenarios. However, if reducing distance to nephrologist care does not alter mortality or hospitalization among remote-dwellers, the cost-effectiveness becomes less attractive. All other one-way sensitivity analyses had negligible effects on the ICUR. Given the low costs of equipping and operating new clinics, and the very attractive ICUR relative to other currently funded interventions, establishing new clinics for remote-dwellers could play an important role in efficiently improving outcomes for patients with CKD. High-quality controlled studies are required to confirm this hypothesis.


Development of a Cre-recombination-based color-switching reporter system for cell fusion detection.
Cell fusion plays a key role in the development and formation of tissues and organs in several organisms. Skeletal myogenesis is assessed in vitro by cell shape and gene and protein expression using immunofluorescence and immunoblotting assays. However, these conventional methods are complex and do not allow for easy time-course observation in living cells. Therefore, this study aimed to develop a Cre recombination-based fluorescent reporter system to monitor cell-cell fusion. We combined green and red fluorescent proteins with a Cre-loxP system to detect syncytium formation using a fluorescent binary switch. This allowed us to visualize mononucleated cells with green fluorescence before fusion and multinucleated syncytia with red fluorescence by conditional expression after cell fusion. The formation of multinuclear myotubes during myogenic differentiation was detected by the change in fluorescence from green to red after Cre-mediated recombination. The distribution of the fluorescence signal correlated with the expression of myogenic differentiation markers. Moreover, red reporter fluorescence intensity was correlated with the number of nuclei contained in the red fluorescent-positive myotubes. We also successfully demonstrated that our fusion monitoring system is applicable to the formation of skeletal muscle myotube and placental syncytiotrophoblast. These results suggest that the color-switching fluorescent reporter system, using Cre-mediated recombination, could be a robust tool used to facilitate the study of cell-to-cell fusion.


Analysis of inhibitory effect of dopamine on carotid body chemoreceptors in cats.
The inhibitory effect of dopamine on carotid body chemoreceptors was studied in anesthetized cats to determine whether it was dependent on changes in blood flow in the vicinity of the receptors. The blood supply to the carotid body was isolated, and flow was controlled with a perfusion pump. Single- or few-fiber recordings were made from the peripheral end of the cut carotid sinus nerve in seven cats. The rate of discharge of 68 chemoreceptor strands increased when flow through the carotid body was stopped. This response was reduced or abolished by dopamine in animals ventilated with either room air (15 strands) or a gas mixture of 95% O2 and 5% CO2 (53 strands). These results suggest that dopamine exerts its inhibitory effect primarily through a direct action on the chemoreceptors rather than by a vasomotor effect in the carotid body.


CLIC4 mediates and is required for Ca2+-induced keratinocyte differentiation.
Keratinocyte differentiation requires integrating signaling among intracellular ionic changes, kinase cascades, sequential gene expression, cell cycle arrest, and programmed cell death. We now show that Cl(-) intracellular channel 4 (CLIC4) expression is increased in both mouse and human keratinocytes undergoing differentiation induced by Ca(2+), serum and the protein kinase C (PKC)-activator, 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Elevation of CLIC4 is associated with signaling by PKCdelta, and knockdown of CLIC4 protein by antisense or shRNA prevents Ca(2+)-induced keratin 1, keratin 10 and filaggrin expression and cell cycle arrest in differentiating keratinocytes. CLIC4 is cytoplasmic in actively proliferating keratinocytes in vitro, but the cytoplasmic CLIC4 translocates to the nucleus in keratinocytes undergoing growth arrest by differentiation, senescence or transforming growth factor beta (TGFbeta) treatment. Targeting CLIC4 to the nucleus of keratinocytes via adenoviral transduction increases nuclear Cl(-) content and enhances expression of differentiation markers in the absence of elevated Ca(2+). In vivo, CLIC4 is localized to the epidermis in mouse and human skin, where it is predominantly nuclear in quiescent cells. These results suggest that CLIC4 participates in epidermal homeostasis through both alterations in the level of expression and subcellular localization. Nuclear CLIC4, possibly by altering the Cl(-) and pH of the nucleus, contributes to cell cycle arrest and the specific gene expression program associated with keratinocyte terminal differentiation.


Collagens and cartilage matrix homeostasis.
At least 27 types of collagen, the products of forty genes, are expressed in the tissues of higher vertebrates. Cartilage has a distinctive collagen phenotype. Two-thirds of the dry weight of adult articular cartilage is collagen. Proteolysis of this collagen framework is integral to the process of cartilage destruction and joint failure in osteoarthritis. Molecular studies are revealing the mechanisms of assembly of cartilage collagen fibrils. The nascent Type II collagen fibril is a heteropolymer, with collagen IX molecules covalently linked to the surface and collagen XI forming a filamentous template at the core, which regulates fibril diameter through its retained N-propeptide domains. This structure presents a challenge to understanding how fibril growth and collagen network maturation are brought about. Proteolytic remodeling, other than that mediated by collagenases, would appear to be involved, but the proteases and molecular mechanisms are still undefined. Valuable insights and predictions on the function of the individual collagen types in cartilage continue to come from the study of skeletal dysplasia syndromes caused by mutations in genes for collagens and associated matrix proteins.


How to find the sensitivity limit for DNA sequencing based on laser-induced fluorescence.
The sensitivity limit for one-lane DNA sequencing based on laser-induced fluorescence is derived by statistical means. A closed-form expression for the sequencing error probability as a function of the signal strength and of the differences in the fluorescence properties of the labels is given. Fluctuations of the signal caused by photodestruction of the molecules cannot be neglected. The expected sequencing error probability is expressed in terms of the number of molecules and the physical parameters of the measurement. This equation can be used in many ways to design and optimize sequencing experiments.


The "perfusion map" of the unipedicled TRAM flap to reduce postoperative partial necrosis.
The unipedicled transverse rectus abdominis musculocutaneous (TRAM) flap is a well-known technique for breast reconstruction. However, it is clinically difficult to evaluate the blood perfusion of the flap in the operating room. A new technique of blood supply evaluation, employing indocyanine green dye (ICG) fluorescence videoangiography has been performed in 10 cases of unipedicled TRAM flap breast reconstruction. In our series, the ICG measurement was demonstrated to be a safe, quick, and accurate technique of flap perfusion analysis. We confirmed the presence of individual pattern ("perfusion map") of the flap perfusion, zone II sometimes not being as well perfused as zone III. In this paper, we present our descriptive findings, and the ICG analysis seems to have a predictive value of unipedicled TRAM flap viability.


Staging of incidentally detected prostate cancer: role of repeat resection, prostate-specific antigen, needle biopsy, and imaging.
Numerous studies have confirmed the distinct biological behavior of two subsets of prostate cancer diagnosed incidentally after either transurethral resection (TURP) or open prostatectomy for presumed benign prostatic hyperplasia (BPH). Focal, low-grade lesions are associated with a low risk for clinical progression and are designated as stage T1a or A1. These cases have traditionally been managed conservatively with close clinical observation. In contrast, multifocal, high-volume, or high-grade tumors are associated with a more aggressive clinical course and are designated as stage T1b or A2. Early definitive intervention is usually advocated for these latter patients. Therefore, accurate pathological assignment to either stage T1a or T1b is crucial for selection of appropriate management options. A variety of methods for staging patients with incidentally detected prostate cancer have been proposed, including detailed histological analysis, repeat TURP or transurethral biopsy, serial prostate-specific antigen (PSA) analysis, and imaging with either transrectal ultrasound (TRUS) or magnetic resonance (MRI) techniques. This article critically examines the clinical utility of these staging modalities for patients with incidentally detected prostate cancer.


SOX9: a useful marker for pancreatic ductal lineage of pancreatic neoplasms.
Previous studies showed that SOX9 plays a critical role in pancreatic ductal development. The aim of this study was to evaluate SOX9 as a marker for pancreatic ductal lineage. SOX9 expression was evaluated by immunohistochemistry in 146 benign pancreas (BP), 136 pancreatic ductal adenocarcinomas, 47 pancreatic intraepithelial neoplasia (PanIN), 21 intraductal papillary mucinous neoplasms (IPMNs), 14 mucinous cystic neoplasms, 10 serous cystadenomas, 39 pancreatic neuroendocrine tumors, 9 acinar cell carcinomas, and 23 solid pseudopapillary neoplasms. Nuclear expression of SOX9 was detected in the centroacinar cells and ductal cells, but not in acinar or endocrine cells in 100% BP. Focal or diffuse SOX9 expression was detected in 100% PanINs, 100% IPMNs, 100% mucinous cystic neoplasms, 100% serous cystadenomas, 89.0% pancreatic ductal adenocarcinomas, 2.6% pancreatic neuroendocrine tumors, 11.1% acinar cell carcinomas, and 0% solid pseudopapillary neoplasms. SOX9 expression was lower in PanIN2 and PanIN3 than in PanIN1 lesions (P < .01). Compared with BP, IPMN had lower SOX9 expression (P < .05). No correlation between SOX9 expression and other clinicopathologic parameters was identified. Our study showed that SOX9 is expressed in centroacinar and ductal epithelial cells of BP and is a useful marker for pancreatic ductal lineage of pancreatic neoplasms.


Does ICP monitoring make a difference in neurocritical care?
Raised intracranial pressure and low cerebral perfusion pressure are associated with ischaemia and poor outcome after brain injury. Therefore, many management protocols target these variables. However, there are no randomized controlled trials that have demonstrated the effectiveness of intracranial pressure-guided care in severely head-injured patients. Observational studies of such therapy have yielded inconsistent results, ranging from decreased mortality to no effect or increased morbidity or mortality. A recent cohort study supports the notion that the possible benefits of intracranial pressure monitoring after traumatic brain injury are small - if present - and would exceed a number needed for the treatment of 16. Furthermore, intracranial pressure monitoring and aggressive management of intracranial pressure and cerebral perfusion pressure have been associated with increased lengths of stay in the neurocritical care unit, conceivable costs and possibly an increased rate of complications. Against this background, there is sufficient clinical equipoise to warrant an adequately powered randomized controlled trial to compare intracranial pressure-guided care with supportive critical care without intracranial pressure monitoring in patients with severe traumatic brain injury. However, the realization of such a trial is likely to be problematic for a number of reasons, not least of which the firmly held biases of many clinicians.


Macadamia Nuts Oil in Nanocream and Conventional Cream as Skin Anti-Aging: A Comparative Study.
The effect of an active substance in a dosage form can be influenced by the different method of preparation. Nanotechnology has been used widely in all aspects including drug delivery system. Nanocream is one of the cosmetic dosage forms to improve the absorption of active substances on the skin. The study was to compare macadamia nuts oil nanocream with conventional cream as skin antiaging dosage form. The formulation of nanocream was consisted of macadamia nuts oil 10%, tween 80, propylene glycol, cetyl alcohol, methyl paraben, propyl paraben and distilled water. The anti-aging was conducted by comparing the nanocream and conventional cream. The evaluation of anti-aging activity was conducted using skin analyzer apparatus. The aging parameters were moisture, evenness, pore size, black spot, and wrinkle of the skin. The data were collected for 4 weeks. All the data obtained were statistically analyzed. Macadamia nut oil nanocream dosage form showed better antiaging activity on the skin compared to conventional cream which were characterized by changes in skin condition on each aging skin parameter such as water content, pore size, melanin, and wrinkles. It is concluded that macadamia nuts oil in nanocream dosage form has better antiaging activity on the skin compared to conventional cream and macadamia nuts oil can be used as an effective skin anti-aging dosage form.


Fisetin Induces Apoptosis Through p53-Mediated Up-Regulation of DR5 Expression in Human Renal Carcinoma Caki Cells.
Fisetin is a natural compound found in fruits and vegetables such as strawberries, apples, cucumbers, and onions. Since fisetin can elicit anti-cancer effects, including anti-proliferation and anti-migration, we investigated whether fisetin induced apoptosis in human renal carcinoma (Caki) cells. Fisetin markedly induced sub-G1 population and cleavage of poly (ADP-ribose) polymerase (PARP), which is a marker of apoptosis, and increased caspase activation. We found that pan-caspase inhibitor (z-VAD-fmk) inhibited fisetin-induced apoptosis. In addition, fisetin induced death receptor 5 (DR5) expression at the transcriptional level, and down-regulation of DR5 by siRNA blocked fisetin-induced apoptosis. Furthermore, fisetin induced p53 protein expression through up-regulation of protein stability, whereas down-regulation of p53 by siRNA markedly inhibited fisetin-induced DR5 expression. In contrast, fisetin induced up-regulation of CHOP expression and reactive oxygen species production, which had no effect on fisetin-induced apoptosis. Taken together, our study demonstrates that fisetin induced apoptosis through p53 mediated up-regulation of DR5 expression at the transcriptional level.


Immune-Related Comorbidities in Childhood-Onset Obsessive Compulsive Disorder: Lifetime Prevalence in the Obsessive Compulsive Disorder Collaborative Genetics Association Study.
Objective: To evaluate the lifetime prevalence of infectious, inflammatory, and autoimmune disorders in a multisite study of probands with childhood-onset obsessive compulsive disorder (OCD) and their first-degree relatives. Methods: Medical questionnaires were completed by 1401 probands and 1045 first-degree relatives in the OCD Collaborative Genetics Association Study. Lifetime prevalence of immune-related diseases was compared with the highest available population estimate and reported as a point estimate with 95% adjusted Wald interval. Worst-episode OCD severity and symptom dimensions were assessed with the Yale-Brown Obsessive Compulsive Scale (YBOCS) and Symptom Checklist (YBOCS-CL). Results: Probands reported higher-than-expected prevalence of scarlet fever (4.0 [3.1-5.2]% vs. 1.0%-2.0%, z = 1.491, p < 0.001, n = 1389), encephalitis or meningitis (1.4 [0.9-2.1]% vs. 0.1%-0.4%, z = 5.913, p < 0.001, n = 1393), rheumatoid arthritis (1.1 [0.6-2.0]% vs. 0.2%-0.4%, z = 3.416, p < 0.001, n = 949) and rheumatic fever (0.6 [0.3-1.2]% vs. 0.1%-0.2%, z = 3.338, p < 0.001, n = 1390), but not systemic lupus erythematosus, diabetes, asthma, multiple sclerosis, psoriasis, or inflammatory bowel disease. First-degree relatives reported similarly elevated rates of scarlet fever, rheumatic fever, and encephalitis or meningitis independent of OCD status. There was no association between worst-episode severity and immune-related comorbidities, although probands reporting frequent ear or throat infections had increased severity of cleaning-/contamination-related symptoms (mean factor score 2.5 ± 0.9 vs. 2.3 ± 1.0, t = 3.183, p = 0.002, n = 822). Conclusion: These data suggest high rates of streptococcal-related and other immune-mediated diseases in patients with childhood-onset OCD and are consistent with epidemiological studies in adults noting familial clustering. Limitations include potential reporting bias and absence of a control group, underscoring the need for further prospective studies characterizing medical and psychiatric disease clusters and their interactions in children. Such studies may ultimately improve our understanding of OCD pathogenesis and aid in the development of adjunctive immune-modulating therapeutic strategies.


Sexual selection driving diversification in jumping spiders.
Theory predicts that speciation rates should be accelerated in organisms undergoing sexual selection. In systems involving female choice, sexual selection acts directly on traits that may be important in prezygotic reproductive isolation, potentially fostering rapid divergence of such traits among allopatric populations. Despite the appeal of this concept, it has proven difficult to document. We provide genetic, behavioral, and simulation data illustrating that the striking and possibly recent divergence in traits of male behavior and morphology among populations of the jumping spider Habronattus pugillis can be attributed to sexual selection. We have found evidence for varying degrees of lower female response and offspring viability among some between-population crosses, consistent with the early stages of speciation. We have developed a gene-tree-based method for comparing phenotypic and genetic data sets to infer selection, and have found robust statistical evidence that directional selection has acted on male traits, by confirming that their rate of fixation exceeds that of neutral mitochondrial genes. Because these traits are apparent targets of female choice, the results indicate that sexual selection is driving divergence of phenotypes potentially crucial to the speciation process.


Genomic characterization of the non-O1/non-O139 Vibrio cholerae strain that caused a gastroenteritis outbreak in Santiago, Chile, 2018.
Vibrio cholerae is a human pathogen, which is transmitted by the consumption of contaminated food or water. V. cholerae strains belonging to the serogroups O1 and O139 can cause cholera outbreaks and epidemics, a severe life-threatening diarrheal disease. In contrast, serogroups other than O1 and O139, denominated as non-O1/non-O139, have been mainly associated with sporadic cases of moderate or mild diarrhea, bacteremia and wound infections. Here we investigated the virulence determinants and phylogenetic origin of a non-O1/non-O139 V. cholerae strain that caused a gastroenteritis outbreak in Santiago, Chile, 2018. We found that this outbreak strain lacks the classical virulence genes harboured by O1 and O139 strains, including the cholera toxin (CT) and the toxin-coregulated pilus (TCP). However, this strain carries genomic islands (GIs) encoding Type III and Type VI secretion systems (T3SS/T6SS) and antibiotic resistance genes. Moreover, we found these GIs are wide distributed among several lineages of non-O1/non-O139 strains. Our results suggest that the acquisition of these GIs may enhance the virulence of non-O1/non-O139 strains that lack the CT and TCP-encoding genes. Our results highlight the pathogenic potential of these V. cholerae strains.


The pediatric trauma center and the inclusive trauma system: Impact on splenectomy rates.
Before 2006, the Delaware Trauma System (DTS) did not include a designated pediatric trauma center (PTC). In 2006, the Delaware Trauma System designated and the American College of Surgeons Committee on Trauma verification/consultation program verified Nemours AI DuPont Hospital for Children, a freestanding children's hospital, as a PTC. We evaluated the impact of the addition of the PTC to the state trauma system on pediatric traumatic splenectomy rates. The study cohort comprised DTS trauma registry recorded children younger than 16 years with spleen injury (ICD-9 codes 865.0-865.9) from January 1998 through December 2012. This cohort was categorized into pre-PTC (1998-2005) and post-PTC (2006-2012) groups. Penetrating injuries were excluded. Comparisons between groups included age, gender, length of stay, organ-specific injury grade, Injury Severity Score, incidence of polytrauma, splenectomy rate, and admitting hospital. Management, operative versus nonoperative, of low grade (Organ Injury Scale [OIS] score, 1-3) and high grade (OIS score, 4-5) were also compared. Pearson's χ analysis was performed for categorical variables. Continuous variables were reported as mean (standard deviation) and compared by Student's t test for independent normally distributed samples. Mann-Whitney U-test was used for non-normally distributed variables. A value of p < 0.05 was considered significant. Of the 231 pediatric spleen injuries, 118 occurred pre-PTC and 113 occurred post-PTC. There were no significant differences in age, gender, length of stay, Injury Severity Score, OIS grade, or incidence of polytrauma. Splenectomy rates decreased from 11% (13 of 118) pre-PTC to 2.7% (3 of 113) post-PTC (p = 0.012). The addition of an American College of Surgeons-verified PTC within an inclusive trauma system that was previously without one was associated with a significant reduction in the rate of blunt trauma-related splenectomy. Integration of a verified PTC is an influential factor in achieving spleen preservation rates equivalent to published American Pediatric Surgery Association benchmarks within a trauma system. Therapeutic study, level IV; epidemiologic study, level III.


The L1-type cell adhesion molecule neuroglian influences the stability of neural ankyrin in the Drosophila embryo but not its axonal localization.
Ankyrins are linker proteins, which connect various membrane proteins, including members of the L1 family of neural cell adhesion molecules, with the submembranous actin-spectrin skeleton. Here we report the cloning and characterization of a second, novel Drosophila ankyrin gene (Dank2) that appears to be the result of a gene duplication event during arthropod evolution. The Drosophila L1-type protein neuroglian interacts with products from both Drosophila ankyrin genes. Whereas the previously described ankyrin gene is ubiquitously expressed during embryogenesis, the expression of Dank2 is restricted to the nervous system in the Drosophila embryo. The absence of neuroglian protein in a neuroglian null mutant line causes decreased levels of Dank2 protein in most neuronal cells. This suggests that neuroglian is important for the stability of Dank2 protein. However, neuroglian is not required for Dank2 axonal localization. In temperature-sensitive neuroglian mutants in which neuroglian protein is mislocated at the restrictive temperature to an intracellular location in the neuronal soma, Dank2 protein can still be detected along embryonic nerve tracts.


Targeted Delivery Systems for Molecular Therapy in Skeletal Disorders.
Abnormalities in the integral components of bone, including bone matrix, bone mineral and bone cells, give rise to complex disturbances of skeletal development, growth and homeostasis. Non-specific drug delivery using high-dose systemic administration may decrease therapeutic efficacy of drugs and increase the risk of toxic effects in non-skeletal tissues, which remain clinical challenges in the treatment of skeletal disorders. Thus, targeted delivery systems are urgently needed to achieve higher drug delivery efficiency, improve therapeutic efficacy in the targeted cells/tissues, and minimize toxicities in non-targeted cells/tissues. In this review, we summarize recent progress in the application of different targeting moieties and nanoparticles for targeted drug delivery in skeletal disorders, and also discuss the advantages, challenges and perspectives in their clinical translation.


Could Sumac Be Effective on COVID-19 Treatment?
Sumac is an herbal product, commonly consumed as a spice and was used for medical treatment for centuries. The phytochemical structure of Sumac was studied extensively, and it was established that the herb contained tannins, polyphenols, flavonoids, organic acids, and essential oils. Various scientific studies demonstrated that Sumac had a free oxygen radical-scavenging effect, a protective effect against liver damage, antihemolytic, leukopenia, and antifibrogenic effects, along with its antiviral, antimicrobial, anti-inflammatory, and antioxidant properties. Recently, several scientific studies described the pathophysiology, clinical course, and the treatment of COVID-19 infection. The examination of the characteristics of COVID-19 infection revealed via the clinical studies suggests that Sumac extract could be useful in the treatment of COVID-19. Given the scientific studies focusing on the beneficial effects of Sumac, the present review aims to provide an encouraging viewpoint to investigate whether Sumac is effective in treating COVID-19 infection.


Fatal muscarinic syndrome after eating wild mushrooms.
Death from mushroom poisoning in Australia is rare and usually due to liver failure produced by Amanita phalloides. We report a 53-year-old woman in Queensland who died from an acute muscarinic syndrome 10 hours after eating mushrooms belonging to the genus Rubinoboletus. To our knowledge, this is the first death in Australia caused by non-amatoxin-producing mushrooms. It highlights the need for awareness of non-amatoxin-producing mushrooms as potentially lethal.


Ruling out Pulmonary Embolism in Patients with (Suspected) COVID-19-A Prospective Cohort Study.
Background Diagnostic strategies for suspected pulmonary embolism (PE) have not been prospectively evaluated in COVID-19 patients. Methods Prospective, multicenter, outcome study in 707 patients with both (suspected) COVID-19 and suspected PE in 14 hospitals. Patients on chronic anticoagulant therapy were excluded. Informed consent was obtained by opt-out approach. Patients were managed by validated diagnostic strategies for suspected PE. We evaluated the safety (3-month failure rate) and efficiency (number of computed tomography pulmonary angiographies [CTPAs] avoided) of the applied strategies. Results Overall PE prevalence was 28%. YEARS was applied in 36%, Wells rule in 4.2%, and "CTPA only" in 52%; 7.4% was not tested because of hemodynamic or respiratory instability. Within YEARS, PE was considered excluded without CTPA in 29%, of which one patient developed nonfatal PE during follow-up (failure rate 1.4%, 95% CI 0.04-7.8). One-hundred seventeen patients (46%) managed according to YEARS had a negative CTPA, of whom 10 were diagnosed with nonfatal venous thromboembolism (VTE) during follow-up (failure rate 8.8%, 95% CI 4.3-16). In patients managed by CTPA only, 66% had an initial negative CTPA, of whom eight patients were diagnosed with a nonfatal VTE during follow-up (failure rate 3.6%, 95% CI 1.6-7.0). Conclusion Our results underline the applicability of YEARS in (suspected) COVID-19 patients with suspected PE. CTPA could be avoided in 29% of patients managed by YEARS, with a low failure rate. The failure rate after a negative CTPA, used as a sole test or within YEARS, was non-negligible and reflects the high thrombotic risk in these patients, warranting ongoing vigilance.


Hypoxia imaging and radiotherapy: bridging the resolution gap.
Oxygen distribution is a major determinant of treatment success in radiotherapy, with well-oxygenated tumour regions responding by up to a factor of three relative to anoxic volumes. Conversely, tumour hypoxia is associated with treatment resistance and negative prognosis. Tumour oxygenation is highly heterogeneous and difficult to measure directly. The recent advent of functional hypoxia imaging modalities such as fluorine-18 fluoromisonidazole positron emission tomography have shown promise in non-invasively determining regions of low oxygen tension. This raises the prospect of selectively increasing dose to hypoxic subvolumes, a concept known as dose painting. Yet while this is a promising approach, oxygen-mediated radioresistance is inherently a multiscale problem, and there are still a number of substantial challenges that must be overcome if hypoxia dose painting is to be successfully implemented. Current imaging modalities are limited by the physics of such systems to have resolutions in the millimetre regime, whereas oxygen distribution varies over a micron scale, and treatment delivery is typically modulated on a centimetre scale. In this review, we examine the mechanistic basis and implications of the radiobiological oxygen effect, the factors influencing microscopic heterogeneity in tumour oxygenation and the consequent challenges in the interpretation of clinical hypoxia imaging (in particular fluorine-18 fluoromisonidazole positron emission tomography). We also discuss dose-painting approaches and outline challenges that must be addressed to improve this treatment paradigm.


SMART--Sunflower Mutant population And Reverse genetic Tool for crop improvement.
Sunflower (Helianthus annuus L.) is an important oilseed crop grown widely in various areas of the world. Classical genetic studies have been extensively undertaken for the improvement of this particular oilseed crop. Pertaining to this endeavor, we developed a "chemically induced mutated genetic resource for detecting SNP by TILLING" in sunflower to create new traits. To optimize the EMS mutagenesis, we first conducted a "kill curve" analysis with a range of EMS dose from 0.5% to 3%. Based on the observed germination rate, a 50% survival rate i.e. LD50, treatment with 0.6% EMS for 8 hours was chosen to generate 5,000 M2 populations, out of which, 4,763 M3 plants with fertile seed set. Phenotypic characterization of the 5,000 M2 mutagenised lines were undertaken to assess the mutagenesis quality and to identify traits of interest. In the M2 population, about 1.1% of the plants showed phenotypic variations. The sunflower TILLING platform was setup using Endo-1-nuclease as mismatch detection system coupled with an eight fold DNA pooling strategy. As proof-of-concept, we screened the M2 population for induced mutations in two genes related to fatty acid biosynthesis, FatA an acyl-ACP thioesterase and SAD the stearoyl-ACP desaturase and identified a total of 26 mutations. Based on the TILLING of FatA and SAD genes, we calculated the overall mutation rate to one mutation every 480 kb, similar to other report for this crop so far. As sunflower is a plant model for seed oil biosynthesis, we anticipate that the developed genetic resource will be a useful tool to identify novel traits for sunflower crop improvement.


A preoperative protocol for the prevention of infection in children with tunnelled right atrial catheters.
The use of central venous lines has come to be widely accepted by children with cancer and their families. However, attendant infection is a cause of considerable morbidity. Coagulase-negative staphylococci, the predominant aerobic species on the skin, are now the commonest cause of catheter-related bacteremia. We introduced a protocol to reduce the colonization of the skin at the catheter insertion site. Antiseptic skin scrubs, with 4% chlorhexidine gluconate, were performed on the neck and anterior chest the night before and again on the morning of the surgical procedure. A single dose of cephalothin (or vancomycin for penicillin-allergic patients) was administered IV immediately before the operation. Compared to the 12 month period prior to initiation of this protocol, the rate of infections (occurring within 30 days of catheter placement) in the 3.5 year period of intervention dropped from 8 to 4.9 per 1,000 catheter days. The proportion of infections that were staphylococcal was reduced from 93 to 63% and the proportion of non-ports removed within 30 days of placement fell from 45 to 0%. Despite these changes, the major contribution to improved infection control appeared to be the use of an increased proportion of ports (a rise from <10 to almost 60%).


The atypical MAPK ERK3 potently suppresses melanoma cell growth and invasiveness.
Mitogen-activated protein kinase 6 (MAPK6) represents an atypical MAPK also known as extracellular signal-regulated kinase 3 (ERK3), which has been shown to play roles in cell motility and metastasis. ERK3 promotes migration and invasion of lung cancer cells and head and neck cancer cells by regulating the expression and/or activity of proteins involved in cancer progression. For instance, ERK3 upregulates matrix metallopeptidases and thereby promotes cancer cell invasiveness, and it phosphorylates tyrosyl-DNA phosphodiesterase 2, thereby enhancing chemoresistance in lung cancer. Here we discovered that ERK3 plays a converse role in melanoma. We observed that BRAF, an oncogenic Ser/Thr kinase, upregulates ERK3 expression levels by increasing both ERK3 messenger RNA levels and protein stability. Interestingly, although BRAF's kinase activity was required for upregulating ERK3 gene transcription, BRAF stabilized ERK3 protein in a kinase-independent fashion. We further demonstrate that ERK3 inhibits the migration, proliferation and colony formation of melanoma cells. In line with this, high level of ERK3 predicted increased survival among patients with melanomas. Taken together, these results indicate that ERK3 acts as a potent suppressor of melanoma cell growth and invasiveness.


Study of the mesenchymal clearance factor and its importance in the middle ear pneumatization process in rat and in man.
An histological study on human and rat embryos and foetuses is presented. In all specimens middle ear pneumatization depended on the correlation between the tubal epithelium development and the mesenchymal absorption. The study of the mastoid pneumatization implies a conceptual development of the chronic otitis media pathogenesis and therefore represents a problem as old as otology itself. It is interesting to recall that the first otologist reached a conceptual idea of chronic otitis media long before the acute one. The initial differentiation was established depending on the duration of the discharge. It is evident that this finding was not sufficient to define chronic otitis, and as R. Bartual (1970) points out, the persistence of a tympanic membrane perforation was later used by the early otologist to differentiate the chronic otitis from the acute one.


Psychosocial factors are the most common contraindications for antiviral therapy at initial evaluation in veterans with chronic hepatitis C.
To understand the degree to which psychosocial factors were preventing veterans with chronic hepatitis C virus from being eligible for antiviral treatment. Nearly 2% of the US population is infected with hepatitis C. Antiviral treatment requires substantial adherence despite challenging side effects. Psychosocial factors, including depression and substance abuse, are clinically recognized contraindications for antiviral treatment. At the Houston Veterans Affairs Medical Center HCV Clinic, we prospectively gathered medical and psychosocial data, as well as treatment disposition data, for consecutively referred patients who had screened positive for hepatitis C by enzyme-linked immunosorbent assay serology. Of the 697 patients referred from September 2000 to May 2001, 580 had chronic hepatitis C. The mean age was 51 years, and 99% were men. Psychosocial contraindications prevented hepatitis C antiviral treatment from being started for 406 (70.0%) patients. These contraindications included alcohol abuse (124, 21.4%), substance abuse (21, 3.6%), and depression (93, 16.0%). Among the medical contraindications were end-stage liver disease (34, 5.9%) and poorly controlled diabetes mellitus (20, 3.4%). Overall, our experience was that a significant portion of hepatitis C patients could not initially be started on antiviral treatment due to psychosocial factors. To make these patients eligible for treatment, future studies of multidisciplinary interventions are required.


The cluster-randomized Quality Initiative in Rectal Cancer trial: evaluating a quality-improvement strategy in surgery.
Following surgery for rectal cancer, two unfortunate outcomes for patients are permanent colostomy and local recurrence of cancer. We tested whether a quality-improvement strategy to change surgical practice would improve these outcomes. Sixteen hospitals were cluster-randomized to the intervention (Quality Initiative in Rectal Cancer strategy) or control (normal practice) arm. Consecutive patients with primary rectal cancer were accrued from May 2002 to December 2004. Surgeons at hospitals in the intervention arm could voluntarily participate by attending workshops, using opinion leaders, inviting a study team surgeon to demonstrate optimal techniques of total mesorectal excision, completing postoperative questionnaires, and receiving audits and feedback. Main outcome measures were hospital rates of permanent colostomy and local recurrence of cancer. A total of 56 surgeons (n = 558 patients) participated in the intervention arm and 49 surgeons (n = 457 patients) in the control arm. The median follow-up of patients was 3.6 years. In the intervention arm, 70% of surgeons participated in workshops, 70% in intraoperative demonstrations and 71% in postoperative questionnaires. Surgeons who had an intraoperative demonstration provided care to 86% of the patients in the intervention arm. The rates of permanent colostomy were 39% in the intervention arm and 41% in the control arm (odds ratio [OR] 0.97, 95% confidence interval [CI] 0.63-1.48). The rates of local recurrence were 7% in the intervention arm and 6% in the control arm (OR 1.06, 95% CI 0.68-1.64). Despite good participation by surgeons, the resource-intense quality-improvement strategy did not reduce hospital rates of permanent colostomy or local recurrence compared with usual practice.


Differential Expression of Vitreous Proteins in Young and Mature New Zealand White Rabbits.
Different anatomical regions have been defined in the vitreous humor including central vitreous, basal vitreous, vitreous cortex, vitreoretinal interface and zonule. In this study we sought to characterize changes in the proteome of vitreous humor (VH) related to compartments or age in New Zealand white rabbits (NZW). Vitreous humor was cryo-collected from young and mature New Zealand white rabbit eyes, and dissected into anterior and posterior compartments. All samples were divided into 4 groups: Young Anterior (YA), Young Posterior (YP), Mature Anterior (MA) and Mature Posterior (MP) vitreous. Tryptic digests of total proteins were analyzed by liquid chromatography followed by tandem mass spectrometry. Spectral count was used to determine the relative protein abundances and identify proteins with statistical differences between compartment and age groups. Western blotting was performed to validate some of the differentially expressed proteins. Our results showed that 231, 375, 273 and 353 proteins were identified in the YA, YP, MA and MP respectively. Fifteen proteins were significantly differentially expressed between YA and YP, and 11 between MA and MP. Carbonic anhydrase III, lambda crystallin, alpha crystallin A and B, beta crystallin B1 and B2 were more abundant in the anterior region, whereas vimentin was less abundant in the anterior region. For comparisons between age groups, 4 proteins were differentially expressed in both YA relative to MA and YP relative to MP. Western blotting confirmed the differential expression of carbonic anhydrase III, alpha crystallin B and beta crystallin B2. The protein profiles of the vitreous humor showed age- and compartment-related differences. This differential protein profile provides a baseline for understanding the vitreous compartmentalization in the rabbit and suggests that further studies profiling proteins in different compartments of the vitreous in other species may be warranted.


Straight stem and threaded cup in patients under 60 years of age: 28.8-30.2 years of follow-up.
The aim of this retrospective observational study of one cohort was to evaluate the long-term outcome in patients younger than 60 years after total hip arthroplasty using a straight uncemented stem and an uncemented threaded cup. Between 1986 and 1987, 75 hips of 75 patients (mean age, 53.35 ± 6.17 years) were consecutively implanted with an Alloclassic Zweymüller/Alloclassic SL stem and an Alloclassic CSF cup. Forty-four patients had died over the last 30 years. The remaining 31 patients (mean age, 82.9 ± 6.4 years) were reinvited for follow-up examinations. Clinical and radiographic evaluations were carried out. At a mean follow-up of 29.5 (28.8-30.2), 4 patients (5.3%) were lost to follow-up. For the endpoint aseptic loosening (defined as the removal of stem or the cup for 2 cases), the overall survival rate is 97.3%. For the endpoint revision for any reason (22 patients), the survival rate is 70.6%. Eleven patients needed an exchange of head and liner, caused by wear. The average time from implantation until change of head and liner was 21.44 years (SD 5.92). Other reasons for revision surgery were septic loosening (3 cases), aseptic loosening of stem and cup (1 case), aseptic loosening of stem (1 case), periprosthetic calcification (2 cases), implant fracture (1 case), periprosthetic fracture (1 case), intraoperative fissure of stem (1 case), and total wear of liner including cup (1 case). The combination of a straight stem (Alloclassic) and a screw cup (CSF) shows excellent results in young patients under the age of 60 at ultra-long-term follow-up at 30 years. Revisions due to wear of the polyethylene liner are more likely than in the older patients.


Glycemic index, cholecystokinin, satiety and disinhibition: is there an unappreciated paradox for overweight women?
The clinical utility of a low glycemic index (LGI) diet for appetite and food intake control is controversial. Complicating the issue are psychological and behavioral influences related to eating. The aim of this study was to investigate the satiety and glycemic response to high GI (HGI) and LGI meals in overweight restrained (R, n=12) and unrestrained (UR, n=10) women. In a randomized crossover study, subjective satiety, cholecystokinin (CCK), glucose, insulin, triacylglyceride (TG) and free fatty acids (FFAs) were measured at defined intervals for 8 h after the participants consumed HGI or LGI test meals. Test meals were matched for energy, energy density, macronutrient content and available carbohydrate, but differed by carbohydrate source; refined grain versus whole grain, respectively. The HGI meal resulted in greater satiety overall, suppressing hunger, desire to eat and prospective consumption compared with the LGI (P<0.01) meal. Plasma CCK was significantly elevated after the HGI meal compared with the LGI meal (P<0.001). Plasma glucose, insulin and TG were higher and FFAs were lower after the HGI meal compared with the LGI meal (P<0001). Dietary restraint did not significantly influence CCK (P=0.14) or subjective satiety (P>0.4); however, an interaction of restraint and disinhibition on CCK was apparent. CCK was blunted in R participants with higher disinhibition scores than UR or R participants with lower disinhibition scores (P<0.05). A LGI diet may not be suitable for optimal satiety and appetite control in overweight women. The relationship between cognitive influences of eating and biobehavioral outcomes requires further investigation.


Sevoflurane postconditioning against cerebral ischemic neuronal injury is abolished in diet-induced obesity: role of brain mitochondrial KATP channels.
Obesity is associated with increased infarct volumes and adverse outcomes following ischemic stroke. However, its effect on anesthetic postconditioning‑induced neuroprotection has not been investigated. The present study examined the effect of sevoflurane postconditioning on focal ischemic brain injury in diet‑induced obesity. Sprague‑Dawley rats were fed a high‑fat diet (HF; 45% kcal as fat) for 12 weeks to develop obesity syndrome. Rats fed a low‑fat diet (LF; 10% kcal as fat) served as controls. The HF or LF‑fed rats were subjected to focal cerebral ischemia for 60 min, followed by 24 h of reperfusion. Postconditioning was performed by exposure to sevoflurane for 15 min immediately at the onset of reperfusion. The involvement of the mitochondrial KATP (mitoKATP) channel was analyzed by the administration of a selective inhibitor of 5‑hydroxydecanoate (5‑HD) prior to sevoflurane postconditioning or by administration of diazoxide (DZX), a mitoKATP channel opener, instead of sevoflurane. The cerebral infarct volume, neurological score and motor coordination were evaluated 24 h after reperfusion. The HF‑fed rats had larger infarct volumes, and lower neurological scores than the LF‑fed rats and also failed to respond to neuroprotection by sevoflurane or DZX. By contrast, sevoflurane and DZX reduced the infarct volumes and improved the neurological scores and motor coordination in the LF‑fed rats. Pretreatment with 5‑HD inhibited sevoflurane‑induced neuroprotection in the LF‑fed rats, whereas it had no effect in the HF‑fed rats. Molecular studies demonstrated that the expression of Kir6.2, a significant mitoKATP channel component, was reduced in the brains of the HF‑fed rats compared with the LF‑fed rats. The results of this study indicate that diet‑induced obesity eliminates the ability of anesthetic sevoflurane postconditioning to protect the brain against cerebral ischemic neuronal injury, most likely due to an impaired brain mitoKATP channel.


Should robotic gastrectomy become a standard surgical treatment option for gastric cancer?
Robotic gastrectomy (RG) using the da Vinci Surgical System for gastric cancer was approved for national medical insurance coverage in Japan in April, 2018, and has been used increasingly since. We reviewed the current evidence on RG, open gastrectomy (OG), and conventional laparoscopic gastrectomy (LG) to identify differences in surgical outcomes between Japan and other countries. Briefly, three independent reviewers systematically reviewed the data collected from a comprehensive literature search by an independent organization and focused on the following nine endpoints: mortality, morbidity, operative time, estimated volume of blood loss, length of postoperative hospital stay, long-term oncologic outcome, quality of life, learning curve, and cost. Overall, the mortality rate of the three approaches did not differ, but RG and LG had less intraoperative blood loss and resulted in a shorter postoperative hospital stay than OG. RG had longer operative times and incurred higher costs than LG and OG. However, in Japan, RG may be more effective than LG and OG for decreasing morbidity. Further studies are needed to establish the specific indications for RG, optimal robotic setup, and dissection methods to best utilize the surgical robot.


A reexamination of the thermoelastic effect in active striated muscle.
Isolated Rana pipiens sartorius muscles at 0degreeC were stimulated via their nerves and small stretches or releases applied during the plateau of the isometric tetanus at lo. Extra heat above the isometric maintenance heat was produced during the drop in tension caused by release and, for very small releases (delta less than or equal to 0.5% lo), was completely reabsorbed during tension recovery. The extra heat was always directly proportional to the tension change. Heat absorption proportional to the tension change was also observed during the increase in tension produced by small stretches in the range 0.5% lo less than or equal to deltal less than or equal to 3.0% lo. The mean heat:tension ratio R in seven experiments was -0.0084, which is within the range of values reported previously by Woledge. In addition, it was found that during tension recovery after small releases of 1.0% lo less than or equal to deltal less than or equal to 3.0% lo the "contractile" component seems able to shorten about 1% lo without producing shortening heat.


Prevalence of elevated hepatic transaminases among Jordanian patients with type 2 diabetes mellitus.
Since the extent of elevation of liver transaminases in type 2 diabetics in Jordan and most of the Middle East is unknown, we estimated the prevalence of elevated liver transaminase levels among patients with type 2 diabetes and determined associated risk factors. This study was performed on 1014 consecutive type 2 diabetic outpatients who attended the National Center for Diabetes, Endocrinology and Genetics in Amman, Jordan. The patients' age ranged between 26-85 years with a mean age of 56.8 (+9.8). Three- hundred and fifty three (54.5%) were males with a median age of 58 years (ranging between 26-82 years), and four hundred and sixty one (45.5%) were females with a median age of 57 years (ranging between 28-85 years). Body mass index, waist circumference, lipid profile, and hepatic transaminase levels were recorded. Ultraonography was performed in those with elevated alanine transaminase levels. Overall, the prevalence of elevated alanine transaminase (ALT) level was 10.4% (n=105) with the gender-wise prevalence being 12.8% (n=71) in men and 7.4% (n=34) in women. The prevalence of elevated aspartate transaminase (AST) levels was 5.4% (n=56) with the gender-wise prevalence being 5.6% (n=31) in men and 5.4% (n=25) in women.. Only 4.5% (n=44) showed elevated levels of both ALT and AST. Male gender (OR=2.35, CI:1.5-3.8) and high waist circumference (OR=1.9, CI:1.2-3.2) were associated with increased risk of elevated ALT levels. Younger patients had a higher tendency to have elevated ALT compared to those over 65 years (OR=12.4 for patients aged 25-45years, and OR=5.8 for those who were 45-65 years old). Non-insulin use was associated with a high odds ratio for elevated ALT levels (OR=1.7, CI: 1.1-2.9). Elevated ALT and AST levels are found in 10.4% and 5.4% of our type 2 diabetic patients respectively. Male gender, younger age, higher waist circumference; as an indicator of central obesity, as well as non insulin use are independent predictors of elevated liver transaminase levels.


During capacitation in bull spermatozoa, actin and PLC-ζ undergo dynamic interactions.
The migration pattern of sperm-specific phospholipase C-ζ (PLC-ζ) was followed and the role of this migration in actin cytoskeleton dynamics was determined. We investigated whether PLC-ζ exits sperm, opening the possibility that PLC-ζ is the 'spermatozoidal activator factor' (SOAF). As capacitation progresses, the highly dynamic actin cytoskeleton bound different proteins to regulate their location and activity. PLC-ζ participation at the start of fertilization was established. In non-capacitated spermatozoa, PLC-ζ is in the perinuclear theca (PT) and in the flagellum, therefore it was decided to determine whether bovine sperm actin interacts with PLC-ζ to direct its relocation as it progresses from non-capacitated (NC) to capacitated (C) and to acrosome-reacted (AR) spermatozoa. PLC-ζ interacted with actin in NC spermatozoa (100%), PLC-ζ levels decreased in C spermatozoa to 32% and in AR spermatozoa to 57% (P < 0.001). The level of actin/PLC-ζ interaction was twice as high in G-actin (P < 0.001) that reflected an increase in affinity. Upon reaching the AR spermatozoa, PLC-ζ was partially released from the cell. It was concluded that actin cytoskeleton dynamics control the migration of PLC-ζ during capacitation and leads to its partial release at AR spermatozoa. It is suggested that liberated PLC-ζ could reach the egg and favour fertilization.


Post-synthetic structural processing in a metal-organic framework material as a mechanism for exceptional CO2/N2 selectivity.
Here we report the synthesis and ceramic-like processing of a new metal-organic framework (MOF) material, [Cu(bcppm)H2O], that shows exceptionally selective separation for CO2 over N2 (ideal adsorbed solution theory, S(ads) = 590). [Cu(bcppm)H2O]·xS was synthesized in 82% yield by reaction of Cu(NO3)2·2.5H2O with the link bis(4-(4-carboxyphenyl)-1H-pyrazolyl)methane (H2bcppm) and shown to have a two-dimensional 4(4)-connected structure with an eclipsed arrangement of the layers. Activation of [Cu(bcppm)H2O] generates a pore-constricted version of the material through concomitant trellis-type pore narrowing (b-axis expansion and c-axis contraction) and a 2D-to-3D transformation (a-axis contraction) to give the adsorbing form, [Cu(bcppm)H2O]-ac. The pore contraction process and 2D-to-3D transformation were probed by single-crystal and powder X-ray diffraction experiments. The 3D network and shorter hydrogen-bonding contacts do not allow [Cu(bcppm)H2O]-ac to expand under gas loading across the pressure ranges examined or following re-solvation. This exceptional separation performance is associated with a moderate adsorption enthalpy and therefore an expected low energy cost for regeneration.


Functionalized triazolopeptoids--a novel class for mitochondrial targeted delivery.
Here we introduce linear 1,4-triazolopeptoids as a novel class of cell penetrating peptidomimetics suitable as organ targeting molecular transporters of bioactive cargo. Repetitive triazole moieties with up to three residues were assembled on solid supports using copper-catalyzed alkyne-azide cycloadditions (CuAAC) in a submonomer approach. Depending on the lipophilicity of their side chain appendages the 1,4-triazolopeptoids showed either endosomal localization or a strong colocalization with the mitochondria of HeLa cells with moderate toxicity. While the basic triazolopeptoids mainly target the neuromast cells in zebrafish embryos, the lipophilic ones colocalize with either cartilage in the jaws and the blood vessel system.


Prophylactic potential of honey and Nigella sativa L. against hospital and community-based SARS-CoV-2 spread: a structured summary of a study protocol for a randomised controlled trial.
Considering the therapeutic potential of honey and Nigella sativa (HNS) in coronavirus disease 2019 (COVID-19) patients, the objective of the study is defined to evaluate the prophylactic role of HNS. The study is a randomized, placebo-controlled, adaptive clinical trial with parallel group design, superiority framework with an allocation ratio of 1:1 among experimental (HNS) and placebo group. An interim analysis will be done when half of the patients have been recruited to evaluate the need to adapt sample size, efficacy, and futility of the trial. All asymptomatic patients with hospital or community based COVID-19 exposure will be screened if they have had 4 days exposure to a confirmed case. Non-pregnant adults with significant exposure level will be enrolled in the study High-risk exposure (<6 feet distance for >10min without face protection) Moderate exposure (<6 feet distance for >10min with face protection) Subjects with acute or chronic infection, COVID-19 vaccinated, and allergy to HNS will be excluded from the study. Recruitment will be done at Shaikh Zayed Post-Graduate Medical Institute, Ali Clinic and Doctors Lounge in Lahore (Pakistan). In this clinical study, patients will receive either raw natural honey (0.5 g) and encapsulated organic Nigella sativa seeds (40 mg) per kg body weight per day or empty capsule with and 30 ml of 5% dextrose water as a placebo for 14 days. Both the natural products will be certified for standardization by Government College University (Botany department). Furthermore, each patient will be given standard care therapy according to version 3.0 of the COVID-19 clinical management guidelines by the Ministry of National Health Services of Pakistan. Primary outcome will be Incidence of COVID-19 cases within 14 days of randomisation. Secondary endpoints include incidence of COVID-19-related symptoms, hospitalizations, and deaths along with the severity of COVID-19-related symptoms till 14th day of randomization. Participants will be randomized into experimental and control groups (1:1 allocation ratio) via the lottery method. There will be stratification based on high risk and moderate risk exposure. Quadruple blinding will be ensured for the participants, care providers and outcome accessors. Data analysts will also be blinded to avoid conflict of interest. Site principal investigator will be responsible for ensuring masking. 1000 participants will be enrolled in the study with 1:1 allocation. The final protocol version 1.4 was approved by institutional review board of Shaikh Zayed Post-Graduate Medical Complex on February 15, 2021. The trial recruitment was started on March 05, 2021, with a trial completion date of February 15, 2022. Clinical trial was registered on February 23, 2021, www.clinicaltrials.gov with registration ID NCT04767087 . The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). With the intention of expediting dissemination of this trial, the conventional formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol. The study protocol has been reported in accordance with the Standard Protocol Items: Recommendations for Clinical Interventional Trials (SPIRIT) guidelines.


A telephone assessment and advice service within an ED physiotherapy clinic: a single-site quality improvement cohort study.
In response to issues with timely access and high non-attendance rates for Emergency Department (ED) physiotherapy, a telephone assessment and advice service was evaluated as part of a quality improvement project. This telehealth option requires minimal resources, with the added benefit of allowing the healthcare professional streamline care. A primary aim was to investigate whether this service model can reduce wait times and non-attendance rates, compared to usual care. A secondary aim was to evaluate service user acceptability. This was a single-site quality improvement cohort study that compares data on wait time to first physiotherapy contact, non-attendance rates and participant satisfaction between patients that opted for a service based on initial telephone assessment and advice, versus routine face-to-face appointments. 116 patients were referred for ED physiotherapy over the 3-month pilot at the ED and out-patient physiotherapy department, XMercy University Hospital, Cork, Ireland. 91 patients (78%) opted for the telephone assessment and advice service, with 40% (n=36) contacting the service. 25 patients (22%) opted for the face-to-face service. Data on wait time and non-attendance rates was gathered using the hospital data reporting system. Satisfaction data was collected on discharge using a satisfaction survey adapted from the General Practice Assessment Questionnaire. Independent-samples t-test or Mann Whitney U Test was utilised depending on the distribution of the data. For categorical data, Chi-Square tests were performed. A level of significance of p ≤ 0.05 was set for this study. Those that contacted the telephone assessment and advice service had a significantly reduced wait time (median 6 days; 3-8 days) compared to those that opted for usual care (median 35 days; 19-39 days) (p ≤ 0.05). There was no significant between-group differences for non-attendance rates or satisfaction. A telephone assessment and advice service may be useful in minimising delays for advice for those referred to ED Physiotherapy for musculoskeleltal problems. This telehealth option appears to be broadly acceptable and since it can be introduced rapidly, it may be helpful in triaging referrals and minimising face-to-face consultations, in line with COVID-19 recommendations. However, a large scale randomised controlled trial is warranted to confirm these findings.


Case report: dextrocardia with situs inversus; two cases presenting differently.
Two patients presented to our centre with dextrocardia and complete situs inversus. The first a 35-year-old presented for the first time with respiratory symptoms but no cardiac symptoms and the second patient (14-year-old) presented with cardiac symptoms. Dextrocardia and situs inversus were confirmed by the chest radiograph, electrocardiograph, and abdominal ultrasound and computerised axial tomography in the two patients. Echocardiography established atrio-ventricular great arteries ventricular concordance and bicuspid aortic valve in the second patient that presented with cardiac symptoms.


The development of cell-initiated degradable hydrogel based on methacrylated alginate applicable to multiple microfabrication technologies.
Biomimetic multicellular complex tissues can be obtained via 3D microfabrication approaches, although the lack of active materials that meet the needs of both molding and the maintenance of cell function is the main bottleneck in their application. The development of 3D cell culture platforms, which are suitable for multiple cell types and various microfabrication technologies, would open the way to the biomimetic construction of multicellular complex tissues. In this study, a MMP-sensitive photocrosslinkable hydrogel system based on methacrylated alginate, which is suitable for the survival of multiple cell types and can be employed in a variety of micro-manufacturing technologies, was designed for the construction of 3D microfeatures to meet both the demands of molding and the requirement for biomimetic function. By employing the characteristics of photocrosslinking and ion crosslinking, the gel system was combined with a variety of microfabrication techniques to construct biomimetic microstructures with a spatial distribution of multiple types of cell. The diversity of shapes and cell types in these assemblies, as well as the maintenance of cell activity, collectively show that the functional MMP-sensitive gel system has the potential to be used as a cell culture platform for the construction of tissue microstructures.


Fine needle aspiration diagnosis of a subcutaneous abscess from Enterobius vermicularis infestation. A case report.
Extraintestinal infestation by Enterobius vermicularis is uncommon. It has been reported to occur in the peritoneal cavity, ovary, fallopian tube, endometrium, lung, liver and urinary tract. Fine needle aspiration diagnosis was made in a case of enterobiasis presenting with a subcutaneous abscess in the natal cleft. Eggs, as well as fragments of cuticle of the adult worm, were found; the morphology of both was best visualized in Papanicolaou-stained smears. Polarizing microscopy highlighted the equally spaced parallel grooves of the cuticle. Fine needle aspiration cytology of subcutaneous abscesses due to enterobiasis can be diagnostic when eggs, or eggs with cuticle, are identified in a suppurative or granulomatous inflammation.


[Natural history of cancer of the kidney: iconographic follow-up].
To describe the natural history of renal cancer based on the images obtained in a case with more than 11 years' follow-up, from the initial stages of the disease to its outcome. The history of a female patient in whom a renal mass had been detected at age 25 years is described. The patient died 11 years later due to metastasis. The images obtained throughout follow-up clearly illustrate the natural course of the disease, which could not be changed because the patient had refused surgery. The observation of a small renal mass in young patients is an indication for early surgery. The natural course of this disease is predictably fatal, its benign behaviour being highly questionable. Because pregnancy does not appear to accelerate the course of the disease, patient assessment can be completed and treatment can be instituted after delivery.


Osteoporosis before lung transplantation: association with low body mass index, but not with underlying disease.
Due to progress in lung transplantation, post-transplantation osteoporosis becomes an important problem. We determined bone mineral density (BMD) in 74 lung transplantation candidates, among them 24 patients with cysticfibrosis, 16 with chronic obstructive pulmonary disease, 14 with pulmonary fibrosis, and 11 with pulmonary hypertension. The mean T score (+/- SD) was -2.6 +/- 1.3 at femoral neck (FN), -2.2 +/- 1.6 at Ward's triangle (WT) and -2.3 +/- 1.5 at lumbar spine (LS). Osteoporosis was found in 61% of the patients at FN, 45% at WT and 50% at LS. Patients with different underlying lung diseases were similarly affected, not only those with cystic fibrosis but also others, including patients with pulmonary hypertension. No association was found between BMD and age, gender, menstrual condition in women and testosterone level in men. A negative correlation was found between chronic glucocorticoid use and T scores. Body mass index correlated positively (p < 0.01) with T scores at any site and the correlation was also significant for the 2 largest subgroups. Loss of lung function (FEV1) also was associated with lower T scores. No correlation was found between BMD and biochemical indices of bone turnover. Multivariate analysis revealed BMI and glucocorticoid use as independent risk factors. We conclude that osteoporosis is a very common condition in patients with end-stage pulmonary disease, independent of the underlying diagnosis. In view of additional bone loss under immunosuppressive treatment after lung transplantation, early diagnosis and prevention of osteoporosis in the pretransplant period should receive high priority.


Arsenite exposure suppresses adipogenesis, mitochondrial biogenesis and thermogenesis via autophagy inhibition in brown adipose tissue.
Arsenite, a trivalent form of arsenic, is an element that occurs naturally in the environment. Humans are exposed to high dose of arsenite through consuming arsenite-contaminated drinking water and food, and the arsenite can accumulate in the human tissues. Arsenite induces oxidative stress, which is linked to metabolic disorders such as obesity and diabetes. Brown adipocytes dissipating energy as heat have emerging roles for obesity treatment and prevention. Therefore, understanding the pathophysiological role of brown adipocytes can provide effective strategies delineating the link between arsenite exposure and metabolic disorders. Our study revealed that arsenite significantly reduced differentiation of murine brown adipocytes and mitochondrial biogenesis and respiration, leading to attenuated thermogenesis via decreasing UCP1 expression. Oral administration of arsenite in mice resulted in heavy accumulation in brown adipose tissue and suppression of lipogenesis, mitochondrial biogenesis and thermogenesis. Mechanistically, arsenite exposure significantly inhibited autophagy necessary for homeostasis of brown adipose tissue through suppression of Sestrin2 and ULK1. These results clearly confirm the emerging mechanisms underlying the implications of arsenite exposure in metabolic disorders.


Anesthesia management of awake craniotomy performed under asleep-awake-asleep technique using laryngeal mask airway: report of two cases.
Asleep-awake-asleep technique of anesthesia is used during awake craniotomy with or without securing airway. We assessed this technique using laryngeal mask airway (LMA) in two patients. Patients underwent awake craniotomy for epilepsy surgery and the removal of a frontotemporal glioma. After anesthesia induction, airway was secured using LMA. Anesthesia was maintained using oxygen, nitrous oxide and sevoflurane, supplemented with an infusion of propofol and remifentanil. Twenty minutes before corticography, anesthesia was discontinued and LMA removed. Both patients were awake and cooperative during the neurological assessment and surgery on eloquent areas. The LMA was reinserted before the closure of the dura and remained in place until the end of surgery. Both patients had no recall of events under anesthesia, although experienced mild pain and discomfort during awake phase of surgery. Both expressed complete satisfaction over the anesthetic management. Asleep-awake-asleep technique using LMA offers airway protection. The painful aspect of surgery can be performed under anesthesia, hence minimizing the duration of stress and pain. Patients remained awake and cooperative throughout the time of neurological testing.


Functional characteristics of neutrophils collected and stored after administration of G-CSF.
Granulocyte transfusion may be used in neutropenic patients with severe bacterial or fungal infections that are unresponsive to antibiotic therapy. However, the inability to store granulocyte concentrates limits their clinical usefulness. Neutrophil chemotaxis and NADPH oxidase activity and the integrity of the neutrophil NADPH oxidase system were examined after apheresis collection and during storage to 48 hours. Neutrophils were mobilized in vivo by G-CSF, collected by apheresis techniques, and stored in apheresis bags in the presence and absence of additional G-CSF. For all experiments, cells were further purified by standard techniques of dextran sedimentation and hypotonic RBC lysis. Neutrophil chemotaxis was preserved to 24 hours of storage but was not affected by the G-CSF added to storage units. The NADPH oxidase system was also preserved as a functioning complex, and both cytosolic proteins and membrane-associated proteins were normal to 48 hours. However, there were divergent responses by intact cells to activating stimuli and reduced oxidase activity in the cell-free system. G-CSF did not appear to significantly affect NADPH oxidase activity or NADPH oxidase system integrity during storage. Neutrophils collected after the administration of G-CSF retained functional and biochemical characteristics for at least 24 hours of storage, which suggests additional effects of G-CSF mobilization beyond enhancing PMN yields and the possibility of storage of these components after collection.