Site-directed mutants of 16S rRNA reveal important RNA domains for KsgA function and 30S subunit assembly.
KsgA is an rRNA methyltransferase important to the process of small subunit biogenesis in bacteria. It is ubiquitously found in all life including archaea and eukarya, where the enzyme is referred to as Dim1. Despite the emergence of considerable data addressing KsgA function over the last several years, details pertaining to RNA recognition are limited, in part because the most accessible substrate for in vitro studies of KsgA is the 900000 Da 30S ribosomal subunit. To overcome challenges imposed by size and complexity, we adapted recently reported techniques to construct in vivo assembled mutant 30S subunits suitable for use in in vitro methyltransferase assays. Using this approach, numerous 16S rRNA mutants were constructed and tested. Our observations indicate that the 790 loop of helix 24 plays an important role in overall catalysis by KsgA. Moreover, the length of helix 45 also is important to catalysis. In both cases loss of catalytic function occurred without an increase in the production of N(6)-methyladenosine, a likely indication that there was no critical reduction in binding strength. Both sets of observations support a "proximity" mechanism of KsgA function. We also report that several of the mutants constructed failed to assemble properly into 30S subunits, while some others did so with reduced efficiency. Therefore, the same technique of generating mutant 30S subunits can be used to study ribosome biogenesis on the whole.


Effect of advancing age on outcomes of deep brain stimulation for Parkinson disease.
Deep brain stimulation (DBS) is a well-established modality for the treatment of advanced Parkinson disease (PD). Recent studies have found DBS plus best medical therapy to be superior to best medical therapy alone for patients with PD and early motor complications. Although no specific age cutoff has been defined, most clinical studies have excluded patients older than 75 years of age. We hypothesize that increasing age would be associated with an increased number of postoperative complications. To evaluate the stepwise effect of increasing age (in 5-year epochs) on short-term complications following DBS surgery. A large, retrospective cohort study was performed using the Thomson Reuters MarketScan national database that examined 1757 patients who underwent DBS for PD during the period from 2000 to 2009. Primary measures examined included hospital length of stay and aggregate and individual complications within 90 days following surgery. Multivariate logistic regression analysis was used to calculate complication-related odds ratios (ORs) for each 5-year age epoch after controlling for covariates. Overall, 132 of 1757 patients (7.5%) experienced at least 1 complication within 90 days, including wound infections (3.6%), pneumonia (2.3%), hemorrhage or hematoma (1.4%), or pulmonary embolism (0.6%). After adjusting for covariates, we found that increasing age (ranging from <50 to 90 years of age) did not significantly affect overall 90-day complication rates (OR, 1.10 per 5-year increase [95% CI, 0.96-1.25]; P = .17). The 2 most common procedure-related complications, hemorrhage (OR, 0.82 [95% CI, 0.63-1.07]; P = .14) and infection (OR, 1.04 [95% CI, 0.87-1.24]; P = .69), did not significantly increase with age. Older patients with PD (>75 years) who were selected to undergo DBS surgery showed a similar 90-day complication risk (including postoperative hemorrhage or infection) compared with younger counterparts. Our findings suggest that age alone should not be a primary exclusion factor for determining candidacy for DBS. Instead, a clear focus on patients with medication-refractory and difficult to control on-off fluctuations with preserved cognition, regardless of age, may allow for an expansion of the traditional therapeutic window.


Surgical correction of vaginal anomalies.
Summary of accurate diagnosis and surgical management of vaginal anomalies. Imperforate hymen, transverse septa, and distal vaginal agenesis present similarly with pain and hematocolpos. Likewise, imperforate hymen, transverse septa, distal vaginal agenesis, and Müllerian aplasia may appear similarly on examination. Imaging should be used to better differentiate the anomaly. Although surgical correction is the mainstay of hymenal anomalies, septa, obstructed hemivaginas, and distal vaginal agenesis, first-line therapy for Müllerian aplasia is progressive dilation. Many surgical techniques are also described but no consensus exists as to the best one. Clinicians should be aware of the appropriate evaluation, differential diagnosis, and management options available for vaginal anomalies.


Fruit and vegetable consumption and prevention of cancer: the Black Churches United for Better Health project.
This study assessed the effects of the Black Churches United for Better Health project on increasing fruit and vegetable consumption among rural African American church members in North Carolina. Ten counties comprising 50 churches were pair matched and randomly assigned to either intervention or delayed intervention (no program until after the follow-up survey) conditions. A multicomponent intervention was conducted over approximately 20 months. A total of 2519 adults (77.3% response rate) completed both the baseline and 2-year follow-up interviews. The 2 study groups consumed similar amounts of fruits and vegetables at baseline. AT the 2-year follow-up, the intervention group consumed 0.85 (SE = 0.12) servings more than the delayed intervention group (P < .0001). The largest increases were observed among people 66 years or older (1 serving), those with education beyond high school (0.92 servings), those widowed or divorced (0.96 servings), and those attending church frequently (1.3 servings). The last improvement occurred among those aged 18 to 37 years and those who were single. The project was a successful model for achieving dietary change among rural African Americans.


Duration of binocular decorrelation in infancy predicts the severity of nasotemporal pursuit asymmetries in strabismic macaque monkeys.
Strabismus in human infants is linked strongly to nasotemporal asymmetries of smooth pursuit, but many features of this co-morbidity are unknown. The purpose of this study was to determine how the duration of early-onset strabismus (or timeliness of repair) affects the severity of pursuit asymmetries in a primate model. Binocular image decorrelation was imposed on infant macaques by fitting them with prism goggles on day 1 of life. The goggles were removed after 3 weeks (n=2), 12 weeks (n=2) or 24 weeks (n=3), emulating surgical repair of strabismus in humans at 3, 12, and 24 months of age, respectively. Two control monkeys wore plano lenses. Several months after the goggles were removed, horizontal smooth pursuit was recorded using binocular search coils and a nasal-bias index (NBI) was calculated. Each animal in the 12- and 24-week groups developed a constant, alternating esotropic strabismus and a nasotemporal asymmetry of pursuit when viewing with either eye. Spatial vision was normal (no amblyopia). The 3-week duration monkeys were indistinguishable from control animals; they had normal eye alignment and symmetric pursuit. In the 12- and 24-week monkeys, the longer the duration of binocular decorrelation, the greater the pursuit asymmetry: for 15 degrees /s target motion, the NBI in the 12-week and 24-week animals was 16x and 22x greater respectively, than that in the 3-week animals (ANOVA, P=0.03). Binocular decorrelation in primates during an early period of fusion development causes permanent smooth pursuit asymmetries when the duration exceeds the equivalent of 3 months in human. These findings support the conclusion that early correction of infantile strabismus promotes normal development of cerebral gaze pathways.


Clinical and mutational heterogeneity of Darier disease in Tunisian families.
To study the mutation spectrum and phenotype-genotype correlation of Darier disease (DD) in Tunisian patients. Case series. Referral center: Department of Dermatology (La Rabta Hospital), Tunis, Tunisia. Eight large Tunisian families with DD, with a total of 23 patients and 9 unaffected family members. Patients were investigated at the clinical, histological, and genetic levels. Families were genotyped with 5 microsatellite markers spanning the ATP2A2 gene. Mutation screening was performed by direct sequencing of the coding region and exon/intron boundaries of the ATP2A2 gene. Typical clinical features of DD were constantly present. Phenotypic variation within and between the studied families was observed. Different neuropsychiatric disorders were seen in 5 families, and various cutaneous and extracutaneous original clinical associations were observed. The haplotype analysis led to the identification of different haplotypes cosegregating with the disease in the studied families. Mutation screening of the ATP2A2 gene revealed 3 recurrent mutations (119-120delAG, R677X, and D702N) and 4 novel variations: 2 missense mutations (G217A and L900R), one microinsertion (2772-2779 ins C), and one microdeletion (1747-1749 del 2T). Our findings provide evidence for clinical and mutational heterogeneity of Tunisian families with DD. No obvious phenotype-genotype correlation was established. To our knowledge, this is the first molecular investigation of DD in the North African population.


Contamination characteristics and source apportionment of methylated PAHs in agricultural soils from Yangtze River Delta, China.
Alkylated PAHs (APAHs) have been shown to be more toxic and persistent than their non-alkylated parent compounds. However, little is known about the extent of soil contamination by these pollutants. To help understand agricultural soil pollution by these compounds at a regional scale, a total of 18 methylated PAHs (MPAHs, a major class of APAHs) in 243 soil samples were analyzed. These soil samples were collected from 11 sites in the Yangtze River Delta (YRD) region, a representative fast developing area in China. The total concentration of MPAHs (∑18MPAHs) ranged from 5.5 to 696.2 ng/g dry soil, with methylnaphthalenes (M-NAPs) and methylphenanthrenes (M-PHEs) accounting for more than 70% of the compositional profile. Relatively high concentrations of ∑18MPAHs were found in Jiaxing and Huzhou areas of Zhejiang province, as well as on the border between the cities of Wuxi and Suzhou. Different MPAH groups showed dissimilar spatial distribution patterns. The spatial distribution of lower molecular weight MPAHs was related to agricultural straw burning and emissions/depositions from industrial activities, whereas that of higher molecular weight MPAHs was much more a function of the total organic carbon (TOC) content of soil. Although coal, biomass (crop straw and wood), and petroleum combustion were identified to be the major emission sources for most of the sampling sites, the areas with relatively severe pollution with ∑18MPAHs resulted from the localized hotspots of petroleum leakage. Isomeric MPAHs with methyl group substituted at 2- (β) position exhibited significantly higher concentrations than those substituted at 1- (α) position. Results of this work help to understand soil pollution by MPAHs, and are useful for designing effective strategies for pollution control so as to ensure food safety in areas with fast economic growth.


Dominant inhibitory Ras mutants selectively inhibit the activity of either cellular or oncogenic Ras.
Two dominant inhibitory Ras mutant proteins were analyzed by microinjection. One, [Asn-17]Ras, had a substitution in the putative Mg(2+)-binding site of Ha-Ras. The other, RAST, had a mutation in a yeast RAS protein that impaired its GTPase activity and increased its affinity for GAP. RAST also had a mutation that blocked its localization to the plasma membrane. In NIH 3T3 cells [Asn-17]Ras inhibited the function of normal Ras much more efficiently than that of oncogenic Ras. In contrast, RAST interfered with the transforming activity of oncogenic Ras more efficiently than that of normal Ras. These conclusions were based on two separate types of analysis. The inhibitory Ras mutant proteins were first microinjected into cells stably transformed either by oncogenic Ras or by high levels of expression of cellular Ras. Results obtained in stably transformed cells were then verified by coinjection of the inhibitory Ras mutant proteins together with transforming concentrations of either oncogenic or normal Ras protein. Whereas RAST was active in soluble form. [Asn-17]Ras required membrane localization for activity. Furthermore, mutations in the GAP/effector-binding domain reduced or eliminated the inhibitory activity of RAST but had no detectable effect on [Asn-17]Ras. These results are consistent with the possibility that [Asn-17]Ras functions by blocking the activation of endogenous Ras proteins, while RAST functions by blocking the ability of activated Ras to stimulate a downstream target within the cells. The properties of RAST suggest that interference with the GAP/effector-binding function of RAS represents a strategy for the preferential inactivation of oncogenic Ras in cells.


Plasma and liver proteomic analysis of 3Z-3-[(1H-pyrrol-2-yl)-methylidene]-1-(1-piperidinylmethyl)-1,3-2H-indol-2-one-induced hepatotoxicity in Wistar rats.
3Z-3-[(1H-pyrrol-2-yl)-methylidene]-1-(1-piperidinylmethyl)-1,3-2H-indol-2-one (Z24), a synthetic anti-angiogenic compound, inhibits the growth and metastasis of certain tumors. Previous works have shown that Z24 induces hepatotoxicity in rodents. We examined the hepatotoxic mechanism of Z24 at the protein level and looked for potential biomarkers. We used 2-DE and MALDI-TOF/TOF MS to analyze alternatively expressed proteins in rat liver and plasma after Z24 administration. We also examined apoptosis in rat liver and measured levels of intramitochondrial ROS and NAD(P)H redox in liver cells. We found that 22 nonredundant proteins in the liver and 11 in the plasma were differentially expressed. These proteins were involved in several important metabolic pathways, including carbohydrate, lipid, amino acid, and energy metabolism, biotransformation, apoptosis, etc. Apoptosis in rat liver was confirmed with the terminal deoxynucleotidyl transferase dUTP-nick end labeling assay. In mitochondria, Z24 increased the ROS and decreased the NAD(P)H levels. Thus, inhibition of carbohydrate aerobic oxidation, fatty acid beta-oxidation, and oxidative phosphorylation is a potential mechanism of Z24-induced hepatotoxicity, resulting in mitochondrial dysfunction and apoptosis-mediated cell death. In addition, fetub protein and argininosuccinate synthase in plasma may be potential biomarkers of Z24-induced hepatotoxicity.


Quantifying the risk of cervical intraepithelial neoplasia in women with unsatisfactory colposcopy results.
To define the risk of high-grade cervical intraepithelial neoplasia (CIN 2,3) among women with unsatisfactory colposcopy results who underwent a loop electrosurgical excision procedure. Loop electrosurgical excision procedures were performed for clinical indications by residents supervised by attending obstetrician-gynecologists at an urban public hospital referral clinic. Specimens obtained between July 1, 1996, and April 30, 2002, were retrieved retrospectively after grading and recording in an institutional database. The endpoint of interest was high-grade cervical disease, a composite of CIN 2, CIN 3, and cancer, in excision specimens. Of 169 evaluable patients, five (3%) had cancer. High-grade disease was found in 6 of 21 patients (29%) without a colposcopic lesion, in 13 of 33 patients (36%) with only koilocytosis on colposcopic biopsy, in 15 of 55 patients (27%) with CIN 1, in 13 of 25 patients (54%) with CIN 2, and in 26 of 35 patients (74%) with CIN 3 (p < 0.001). High-grade disease was associated with the grade of referral cytologic results, cytologic analysis repeated at colposcopy, and colposcopic biopsy (p < 0.001 for all). Limiting excision to women with cytologic results at the time of colposcopy read as atypical squamous cells of undetermined significance or worse yielded a high-grade disease prevalence of 12%, with a sensitivity of 92%, specificity 46%, negative predictive value 88%, and positive predictive value 56%. Referral cytologic results, colposcopic biopsy, age, and endocervical curettage results did not seem to identify women at low risk for high-grade disease. Women with negative cytologic results at the time of colposcopy have a low risk for high-grade disease and may avoid a loop electrosurgical excision procedure despite unsatisfactory colposcopy.


Leupaxin negatively regulates B cell receptor signaling.
The role of the paxillin superfamily of adaptor proteins in B cell antigen receptor (BCR) signaling has not been studied previously. We show here that leupaxin (LPXN), a member of this family, was tyrosine-phosphorylated and recruited to the plasma membrane of human BJAB lymphoma cells upon BCR stimulation and that it interacted with Lyn (a critical Src family tyrosine kinase in BCR signaling) in a BCR-induced manner. LPXN contains four leucine-rich sequences termed LD motifs, and serial truncation and specific domain deletion of LPXN indicated that its LD3 domain is involved in the binding of Lyn. Of a total of 11 tyrosine sites in LPXN, we mutated Tyr(22), Tyr(72), Tyr(198), and Tyr(257) to phenylalanine and demonstrated that LPXN was phosphorylated by Lyn only at Tyr(72) and that this tyrosine site is proximal to the LD3 domain. The overexpression of LPXN in mouse A20 B lymphoma cells led to the suppression of BCR-induced activation of JNK, p38 MAPK, and, to a lesser extent, Akt, but not ERK and NFkappaB, suggesting that LPXN can selectively repress BCR signaling. We further show that LPXN suppressed the secretion of interleukin-2 by BCR-activated A20 B cells and that this inhibition was abrogated in the Y72F LPXN mutant, indicating that the phosphorylation of Tyr(72) is critical for the biological function of LPXN. Thus, LPXN plays an inhibitory role in BCR signaling and B cell function.


Safety of bazedoxifene in a randomized, double-blind, placebo- and active-controlled Phase 3 study of postmenopausal women with osteoporosis.
We report the safety findings from a 3-year phase 3 study (NCT00205777) of bazedoxifene, a novel selective estrogen receptor modulator under development for the prevention and treatment of postmenopausal osteoporosis. Healthy postmenopausal osteoporotic women (N = 7,492; mean age, 66.4 years) were randomized to daily doses of bazedoxifene 20 or 40 mg, raloxifene 60 mg, or placebo for 3 years. Safety and tolerability were assessed by adverse event (AE) reporting and routine physical, gynecologic, and breast examination. Overall, the incidence of AEs, serious AEs, and discontinuations due to AEs in the bazedoxifene groups was not different from that seen in the placebo group. The incidence of hot flushes and leg cramps was higher with bazedoxifene or raloxifene compared with placebo. The rates of cardiac disorders and cerebrovascular events were low and evenly distributed among groups. Venous thromboembolic events, primarily deep vein thromboses, were more frequently reported in the active treatment groups compared with the placebo group; rates were similar with bazedoxifene and raloxifene. Bazedoxifene showed a neutral effect on the breast and an excellent endometrial safety profile. The incidence of fibrocystic breast disease was lower with bazedoxifene 20 and 40 mg versus raloxifene or placebo. Reductions in total and low-density lipoprotein levels and increases in high-density lipoprotein levels were seen with bazedoxifene versus placebo; similar results were seen with raloxifene. Triglyceride levels were similar among groups. Bazedoxifene showed a favorable safety and tolerability profile in women with postmenopausal osteoporosis. NCT00205777; Trial registration date: September 16, 2005.


Grit: what is it and why does it matter in medicine?
Grit refers to the combination of passion and perseverance for long-term goals. Grit has emerged as a recent topic of interest within the medical community. With ever-increasing rates of burnout and psychological distress, increasing attention has been directed towards modulatory or protective factors for these deleterious outcomes. Grit has been studied in regard to a variety of outcomes and variables in medicine. This article reviews the current literature on grit in medicine and summarises the current research on grit and performance metrics, personality characteristics, longitudinal progression, psychological well-being, diversity, equity and inclusion, burnout and residency attrition. While there is inconclusive evidence on the influence of grit on performance metrics in medicine, research consistently demonstrates a positive correlation between grit and psychological well-being and a negative correlation between grit and burnout. After discussing some of the inherent limitations of this type of research, this article suggests some possible implications and future areas for research and their potential role in cultivating psychologically healthy physicians and promoting successful careers in medicine.


Optimizing carbon storage and biodiversity protection in tropical agricultural landscapes.
With the rapidly expanding ecological footprint of agriculture, the design of farmed landscapes will play an increasingly important role for both carbon storage and biodiversity protection. Carbon and biodiversity can be enhanced by integrating natural habitats into agricultural lands, but a key question is whether benefits are maximized by including many small features throughout the landscape ('land-sharing' agriculture) or a few large contiguous blocks alongside intensive farmland ('land-sparing' agriculture). In this study, we are the first to integrate carbon storage alongside multi-taxa biodiversity assessments to compare land-sparing and land-sharing frameworks. We do so by sampling carbon stocks and biodiversity (birds and dung beetles) in landscapes containing agriculture and forest within the Colombian Chocó-Andes, a zone of high global conservation priority. We show that woodland fragments embedded within a matrix of cattle pasture hold less carbon per unit area than contiguous primary or advanced secondary forests (>15 years). Farmland sites also support less diverse bird and dung beetle communities than contiguous forests, even when farmland retains high levels of woodland habitat cover. Landscape simulations based on these data suggest that land-sparing strategies would be more beneficial for both carbon storage and biodiversity than land-sharing strategies across a range of production levels. Biodiversity benefits of land-sparing are predicted to be similar whether spared lands protect primary or advanced secondary forests, owing to the close similarity of bird and dung beetle communities between the two forest classes. Land-sparing schemes that encourage the protection and regeneration of natural forest blocks thus provide a synergy between carbon and biodiversity conservation, and represent a promising strategy for reducing the negative impacts of agriculture on tropical ecosystems. However, further studies examining a wider range of ecosystem services will be necessary to fully understand the links between land-allocation strategies and long-term ecosystem service provision.


Cyclophosphamide boluses induce micronuclei expression in buccal mucosa cells of patients with systemic lupus erythematosus independent of cytochrome P450 2D6 status.
To assess the effect of cyclophosphamide (CYC) and the influence of CYP2D6 polymorphism on micronuclei expression in patients with systemic lupus erythematosus (SLE). Micronuclei (MN) assays were performed in oral mucosa sweeps of 49 patients with SLE, receiving and not receiving CYC therapy, and 43 healthy controls. In patients receiving CYC the MN assay was repeated 14 days after CYC administration. SLE patients underwent determination of the CYP2D6 allele expressed in peripheral blood by polymerase chain reaction amplification. These results were compared with CYP2D6 distribution in a healthy population from the same geographic area. The results were analyzed by Mann-Whitney U and chi-square tests. Twenty-four patients with SLE received CYC boluses. The mean age of SLE patients was 32 +/- 10 years; mean disease duration was 7 +/- 6 years. Basal expression of MN was similar in the groups investigated and increased significantly in those exposed to CYC (CYC group 44%, no CYC group 5%, healthy controls 9%; p = 0.001). We found no association between the CYP2D6 allele expressed and MN induction. Poor metabolizers were overrepresented in SLE subjects as compared with 139 healthy controls (p < 0.05). Genotoxicity, as assessed by the MN assay, is increased in patients with SLE after CYC boluses. The CYP2D6 allele expressed in SLE patients does not have a role in CYC induced chromosomal injury.


Imprecise Data and Their Impact on Translational Research in Medicine.
The medical field expects from big data essentially two main results: the ability to build predictive models and the possibility of applying them to obtain accurate patient risk profiles and/or health trajectories. Note that the paradigm of precision has determined that similar challenges need to be faced in both population and individualized studies, namely the need of assembling, integrating, modeling, and interpreting data from a variety of information sources and scales potentially influencing disease from onset to progression. In many cases, data require computational treatment through solutions for otherwise intractable problems. However, as precision medicine remains subject to a substantial amount of data imprecision and lack of translational impact, a revision of methodological inference approaches is needed. Both the relevance and the usefulness of such revision crucially deal with the assimilation of data features dynamically interconnected.


In vivo persistence, tumor localization, and antitumor activity of CAR-engineered T cells is enhanced by costimulatory signaling through CD137 (4-1BB).
Human T cells engineered to express a chimeric antigen receptor (CAR) specific for folate receptor-α (FRα) have shown robust antitumor activity against epithelial cancers in vitro but not in the clinic because of their inability to persist and home to tumor in vivo. In this study, CARs were constructed containing a FRα-specific scFv (MOv19) coupled to the T-cell receptor CD3ζ chain signaling module alone (MOv19-ζ) or in combination with the CD137 (4-1BB) costimulatory motif in tandem (MOv19-BBζ). Primary human T cells transduced to express conventional MOv19-ζ or costimulated MOv19-BBζ CARs secreted various proinflammatory cytokines, and exerted cytotoxic function when cocultured with FRα(+) tumor cells in vitro. However, only transfer of human T cells expressing the costimulated MOv19-BBζ CAR mediated tumor regression in immunodeficient mice bearing large, established FRα(+) human cancer. MOv19-BBζ CAR T-cell infusion mediated tumor regression in models of metastatic intraperitoneal, subcutaneous, and lung-involved human ovarian cancer. Importantly, tumor response was associated with the selective survival and tumor localization of human T cells in vivo and was only observed in mice receiving costimulated MOv19-BBζ CAR T cells. T-cell persistence and antitumor activity were primarily antigen-driven; however, antigen-independent CD137 signaling by CAR improved T-cell persistence but not antitumor activity in vivo. Our results show that anti-FRα CAR outfitted with CD137 costimulatory signaling in tandem overcome issues of T-cell persistence and tumor localization that limit the conventional FRα T-cell targeting strategy to provide potent antitumor activity in vivo.


Ultrathin Lubricant-Infused Vertical Graphene Nanoscaffolds for High-Performance Dropwise Condensation.
Lubricant-infused surfaces (LIS) are highly efficient in repelling water and constitute a very promising family of materials for condensation processes occurring in a broad range of energy applications. However, the performance of LIS in such processes is limited by the inherent thermal resistance imposed by the thickness of the lubricant and supporting surface structure, as well as by the gradual depletion of the lubricant over time. Here, we present an ultrathin (∼70 nm) and conductive LIS architecture, obtained by infusing lubricant into a vertically grown graphene nanoscaffold on copper. The ultrathin nature of the scaffold, combined with the high in-plane thermal conductivity of graphene, drastically minimize earlier limitations, effectively doubling the heat transfer performance compared to a state-of-the-art CuO LIS surface. We show that the effect of the thermal resistance to the heat transfer performance of a LIS surface, although often overlooked, can be so detrimental that a simple nanostructured CuO surface can outperform a CuO LIS surface, despite filmwise condensation on the former. The present vertical graphene LIS is also found to be resistant to lubricant depletion, maintaining stable dropwise condensation for at least 24 h with no significant change of advancing contact angle and contact angle hysteresis. The lubricant consumed by the vertical graphene LIS is 52.6% less than that of the existing state-of-the-art CuO LIS, also making the fabrication process more economical.


Expression of runtB is modulated during chondrocyte differentiation.
The runt locus in Drosophila encodes a nuclear protein involved in embryo segmentation, sex determination/X dosage compensation, and neurogenesis. runt homologues have been identified in higher vertebrates. The encoded proteins share a domain of 128 amino acids called the runt domain. It has been reported that this domain mediates DNA binding and heterodimerization. Here, we analyze runtB expression during chondrocyte differentiation "in vitro" and "in vivo." We have first isolated, from a chondrocyte library, a cDNA clone coding for a runtB chicken homologue and containing a complete open reading frame. The predicted protein product is 84% identical to the mouse PEBP2alphaB2 isoform. By RT-PCR analysis we have also cloned the chicken cDNA fragment coding for delta alphaB2, the exon sequence included in the B1 isoform mRNA. On Northern blot analysis of cultured chondrocytes, runtB mRNA levels increase dramatically with the transition from stage 0 (dedifferentiated) to stages I and II (hypertrophic chondrocytes). Moreover, runt polypeptides were demonstrated in chondrocytes both in vivo and in vitro. These results suggest that runt plays a role in chondrogenic differentiation.


Minimally invasive coronary artery bypass grafting versus stenting for patients with proximal left anterior descending coronary artery disease.
The purpose of this study was to compare the clinical outcomes of stenting and minimally invasive coronary artery bypass grafting (MIDCAB) in patients with proximal left anterior descending (LAD) coronary artery disease. The Patency, Outcome, Economics of Minimally invasive direct coronary bypass (POEM) study demonstrated that MIDCAB had similar safety and long-term efficacy for LAD revascularization compared with conventional coronary artery bypass grafting. Although LAD stenting is superior to conventional balloon angioplasty, whether it is comparable to MIDCAB is not known. We identified a matched population of 429 consecutive patients with 1-vessel disease who underwent elective proximal LAD stenting and compared their clinical outcomes with those of the 152 patients in the MIDCAB group of the POEM study. The in-hospital event rate was similar in both groups, except for a shorter length of hospital stay with LAD stenting compared with MIDCAB (2.68 vs 4.07 days, p <0.0001). At 6-month follow-up, the incidence of death and Q-wave myocardial infarction or that of cerebrovascular accident was not significantly different between these 2 groups. However, target vessel revascularization was significantly higher with LAD stenting than MIDCAB (13.3% vs 6.6%, p = 0.045). In the subgroup of patients without diabetes, all clinical events were similar in both groups, and the benefit of a shorter hospital stay associated with stenting was maintained. Compared with MIDCAB, LAD stenting is associated with higher repeat revascularization rates but offers the advantage of shorter hospitalization. For nondiabetics with proximal LAD disease, stenting may be the revascularization strategy of choice.


Real-time motion onset recognition for robot-assisted gait rehabilitation.
Many patients with neurological movement disorders fear to fall while performing postural transitions without assistance, which prevents them from participating in daily life. To overcome this limitation, multi-directional Body Weight Support (BWS) systems have been developed allowing them to perform training in a safe environment. In addition to overground walking, these innovative/novel systems can assist patients to train many more gait-related tasks needed for daily life under very realistic conditions. The necessary assistance during the users' movements can be provided via task-dependent support designs. One remaining challenge is the manual switching between task-dependent supports. It is error-prone, cumbersome, distracts therapists and patients, and interrupts the training workflow. Hence, we propose a real-time motion onset recognition model that performs automatic support switching between standing-up and sitting-down transitions and other gait-related tasks (8 classes in total). To predict the onsets of the gait-related tasks, three Inertial Measurement Units (IMUs) were attached to the sternum and middle of outer thighs of 19 controls without neurological movement disorders and two individuals with incomplete Spinal Cord Injury (iSCI). The data of IMUs obtained from different gait tasks was sent synchronously to a real-time data acquisition system through a custom-made Bluetooth-EtherCAT gateway. In the first step, data was applied offline for training five different classifiers. The best classifier was chosen based on F1-score results of a Leave-One-Participant-Out Cross-Validation (LOPOCV), which is an unbiased way of testing. In a final step, the chosen classifier was tested in real time with an additional control participant to demonstrate feasibility for real-time classification. Testing five different classifiers, the best performance was obtained in a single-layer neural network with 25 neurons. The F1-score of [Formula: see text] and [Formula: see text] are achieved on testing using LOPOCV and test data ([Formula: see text], participants = 20), respectively. Furthermore, the results from the implemented real-time classifier were compared with the offline classifier and revealed nearly identical performance (difference = [Formula: see text]). A neural network classifier was trained for identifying the onset of gait-related tasks in real time. Test data showed convincing performance for offline and real-time classification. This demonstrates the feasibility and potential for implementing real-time onset recognition in rehabilitation devices in future.


Pilot test of dosage effects in HEXORR II for robotic hand movement therapy in individuals with chronic stroke.
Impaired use of the hand in functional tasks remains difficult to overcome in many individuals after a stroke. This often leads to compensation strategies using the less-affected limb, which allows for independence in some aspects of daily activities. However, recovery of hand function remains an important therapeutic goal of many individuals, and is often resistant to conventional therapies. In prior work, we developed HEXORR I, a robotic device that allows practice of finger and thumb movements with robotic assistance. In this study, we describe modifications to the device, now called HEXORR II, and a clinical trial in individuals with chronic stroke. Fifteen individuals with a diagnosis of chronic stroke were randomized to 12 or 24 sessions of robotic therapy. The sessions involved playing several video games using thumb and finger movement. The robot applied assistance to extension movement that was adapted based on task performance. Clinical and motion capture evaluations were performed before and after training and again at a 6-month followup. Fourteen individuals completed the protocol. Fugl-Meyer scores improved significantly at the 6 month time point compared to baseline, indicating reductions in upper extremity impairment. Flexor hypertonia (Modified Ashworth Scale) also decreased significantly due to the intervention. Motion capture found increased finger range of motion and extension ability after the intervention that continued to improve during the followup period. However, there was no change in a functional measure (Action Research Arm Test). At the followup, the high dose group had significant gains in hand displacement during a forward reach task. There were no other significant differences between groups. Future work with HEXORR II should focus on integrating it with functional task practice and incorporating grip and squeezing tasks.


Clinical importance of risk variants in the dihydropyrimidine dehydrogenase gene for the prediction of early-onset fluoropyrimidine toxicity.
We investigated the clinical relevance of dihydropyrimidine dehydrogenase gene (DPYD) variants to predict severe early-onset fluoropyrimidine (FP) toxicity, in particular of a recently discovered haplotype hapB3 and a linked deep intronic splice site mutation c.1129-5923C>G. Selected regions of DPYD were sequenced in prospectively collected germline DNA of 500 patients receiving FP-based chemotherapy. Associations of DPYD variants and haplotypes with hematologic, gastrointestinal, infectious, and dermatologic toxicity in therapy cycles 1-2 and resulting FP-dose interventions (dose reduction, therapy delay or cessation) were analyzed accounting for clinical and demographic covariates. Fifteen additional cases with toxicity-related therapy delay or cessation were retrospectively examined for risk variants. The association of c.1129-5923C>G/hapB3 (4.6% carrier frequency) with severe toxicity was replicated in an independent prospective cohort. Overall, c.1129-5923G/hapB3 carriers showed a relative risk of 3.74 (RR, 95% CI = 2.30-6.09, p = 2 × 10(-5)) for severe toxicity (grades 3-5). Of 31 risk variant carriers (c.1129-5923C>G/hapB3, c.1679T>G, c.1905+1G>A or c.2846A>T), 11 (all with c.1129-5923C>G/hapB3) experienced severe toxicity (15% of 72 cases, RR = 2.73, 95% CI = 1.61-4.63, p = 5 × 10(-6)), and 16 carriers (55%) required FP-dose interventions. Seven of the 15 (47%) retrospective cases carried a risk variant. The c.1129-5923C>G/hapB3 variant is a major contributor to severe early-onset FP toxicity in Caucasian patients. This variant may substantially improve the identification of patients at risk of FP toxicity compared to established DPYD risk variants (c.1905+1G>A, c.1679T>G and c.2846A>T). Pre-therapeutic DPYD testing may prevent 20-30% of life-threatening or lethal episodes of FP toxicity in Caucasian patients.


Precision optical path alignment system for point diffraction interferometer based on image information.
In a point diffraction interferometer, the existence of alignment error between an objective convergent spot and a diffraction pinhole can lead to wavefront error and diffraction intensity reduction. Meanwhile, the contrast of the point diffraction interferograms probably decreases in this procedure. These changes will have significant influence on its inspection precision. A precise alignment system of an optical path for a point diffraction interferometer is proposed in this paper. First, diffraction theory is used to analyze the mathematical relationships of alignment error to diffraction wavefront error and numerical aperture and wavefront error to pinhole size. Then, according to the requirement, the scheme of an optical path alignment system is designed. In this stage, alignment images as well as intensities of a reflected and diffracted beam from the point diffraction plate will be acquired. In addition, an image processing algorithm for measuring alignment error is designed, and a mathematical model between quantities of measurement and control is constructed. Finally, implementation and experiment of this method are also introduced. Misalignment situations, including lateral translation, longitudinal defocus, and tilt error, are well eliminated, and the quality of interferograms is also improved. From the results, it can be concluded that the system is of desirable precision and efficiency.


Direct synthesis of an heterometallic {Mn(II)3Cr(III)4} wheel by decomposition of Reineckes salt.
A new heterometallic Mn/Cr molecular wheel complex has been synthesized using zerovalent manganese, Reineckes salt, ammonium thiocyanate and triethanolamine (H(3)tea) as starting materials. The compound [Mn(3)Cr(4)(NCS)(6)(Htea)(6)] (1) has been characterized in terms of its electrochemical, IR and EPR spectroscopic as well as its magnetic properties. The magnetic susceptibility and magnetization data were treated simultaneously during the optimization routine and resulted in J(Mn-Cr) = +0.43 cm(-1), J(Cr-Cr) = -4.75 cm(-1), J(Mn-Mn) = +1.78 cm(-1), g(eff) = 1.878 with the ground state S = 15/2.


Ethylene-promoted adventitious rooting and development of cortical air spaces (aerenchyma) in roots may be adaptive responses to flooding in Zea mays L.
The roots and stem base of intact, 10 day old maize (Zea mays L. cv. LG11) plants, grown in nutrient solution, were continuously aerated either with ethylene (5 μl l(-1)) in air or with air alone. Ethylene treatment hastened the emergence of adventitious (nodal) roots from the base of the shoot, but slowed their subsequent extension. Ethylene also promoted the collapse of cells in the cortex of these roots, with lysigenous development of prominent air spaces (aerenchyma). Non-aeration of the nutrient solution caused endogenously produced ethylene to accumulate in the roots, and stimulated both the emergence of adventitious roots and the formation of cortical air spaces in them. With non-aeration the concentration of oxygen did not fall below 1% in the equilibrium gas phase (air=20.8%). Complete deoxygenation of the nutrient solution, produced by passing oxygen-free nitrogen gas, prevented both air space formation and the evolution of ethylene by root segments.These results suggest that adventitious rooting and cortical air space formation in nodal roots in Zea mays may be stimulated by enhanced concentrations of endogenous ethylene arising either from entrapment of the gas by unstirred water layers around the roots and/or by increased biosynthesis. These responses are considered conducive to survival in waterlogged soil.


Expression of varicella-zoster virus-related antigens in biochemically transformed cells.
L(O)cl 3 and L(K)cl 1 cells, biochemically transformed by varicella-zoster virus (VZV), were labelled with L-[35S]methionine and [32P]orthophosphate. Cell extracts were immunoprecipitated with anti-VZV monkey serum and analysed by SDS-polyacrylamide gel electrophoresis. Four polypeptides of apparent mol. wt. 135000 (135K), 48K, 44K and 35K were detected in the L-[35S]methionine-labelled extracts, and, of these, the 35K band showed marked intensity. However, this band was not detected in extracts from cells infected with a VZV tk- strain (Kanno strain). Also, the 35K polypeptide showed very low intensity when immunoprecipitated from extracts of transformed cells grown in non-selective (NS) medium, i.e. cells that had a very low thymidine kinase (tk) activity. In the case of [32P]orthophosphate-labelled cells, polypeptides of apparent mol. wt. 180K, 81K, 48K, 44K and 37K were obtained. In both instances, the 44K polypeptide was not immunoprecipitated from L(K)cl 1 cell extracts. From our data it is postulated that the expression of the 35K polypeptide is correlated with the VZV-specific tk activity of the cells.


Genome editing retraces the evolution of toxin resistance in the monarch butterfly.
Identifying the genetic mechanisms of adaptation requires the elucidation of links between the evolution of DNA sequence, phenotype, and fitness1. Convergent evolution can be used as a guide to identify candidate mutations that underlie adaptive traits2-4, and new genome editing technology is facilitating functional validation of these mutations in whole organisms1,5. We combined these approaches to study a classic case of convergence in insects from six orders, including the monarch butterfly (Danaus plexippus), that have independently evolved to colonize plants that produce cardiac glycoside toxins6-11. Many of these insects evolved parallel amino acid substitutions in the α-subunit (ATPα) of the sodium pump (Na+/K+-ATPase)7-11, the physiological target of cardiac glycosides12. Here we describe mutational paths involving three repeatedly changing amino acid sites (111, 119 and 122) in ATPα that are associated with cardiac glycoside specialization13,14. We then performed CRISPR-Cas9 base editing on the native Atpα gene in Drosophila melanogaster flies and retraced the mutational path taken across the monarch lineage11,15. We show in vivo, in vitro and in silico that the path conferred resistance and target-site insensitivity to cardiac glycosides16, culminating in triple mutant 'monarch flies' that were as insensitive to cardiac glycosides as monarch butterflies. 'Monarch flies' retained small amounts of cardiac glycosides through metamorphosis, a trait that has been optimized in monarch butterflies to deter predators17-19. The order in which the substitutions evolved was explained by amelioration of antagonistic pleiotropy through epistasis13,14,20-22. Our study illuminates how the monarch butterfly evolved resistance to a class of plant toxins, eventually becoming unpalatable, and changing the nature of species interactions within ecological communities2,6-11,15,17-19.


"Reproducible" Research in Mathematical Sciences Requires Changes in our Peer Review Culture and Modernization of our Current Publication Approach.
The nature of scientific research in mathematical and computational biology allows editors and reviewers to evaluate the findings of a scientific paper. Replication of a research study should be the minimum standard for judging its scientific claims and considering it for publication. This requires changes in the current peer review practice and a strict adoption of a replication policy similar to those adopted in experimental fields such as organic synthesis. In the future, the culture of replication can be easily adopted by publishing papers through dynamic computational notebooks combining formatted text, equations, computer algebra and computer code.


Movement to the Clinic of Soluble Epoxide Hydrolase Inhibitor EC5026 as an Analgesic for Neuropathic Pain and for Use as a Nonaddictive Opioid Alternative.
This report describes the development of an orally active analgesic that resolves inflammation and neuropathic pain without the addictive potential of opioids. EC5026 acts on the cytochrome P450 branch of the arachidonate cascade to stabilize epoxides of polyunsaturated fatty acids (EpFA), which are natural mediators that reduce pain, resolve inflammation, and maintain normal blood pressure. EC5026 is a slow-tight binding transition-state mimic that inhibits the soluble epoxide hydrolase (sEH) at picomolar concentrations. The sEH rapidly degrades EpFA; thus, inhibiting sEH increases EpFA in vivo and confers beneficial effects. This mechanism addresses disease states by shifting endoplasmic reticulum stress from promoting cellular senescence and inflammation toward cell survival and homeostasis. We describe the synthesis and optimization of EC5026 and its development through human Phase 1a trials with no drug-related adverse events. Additionally, we outline fundamental work leading to discovery of the analgesic and inflammation-resolving CYP450 branch of the arachidonate cascade.


Tumor necrosis factor alpha (TNF-alpha) and sepsis: evidence for a role in host defense.
Serum levels of TNF-alpha were evaluated in 29 patients with sepsis, using TNF-alpha sensitive L929 cells (sensitivity = 15 pg/ml). Blood samples were collected serially at the first 24-36 h of symptoms. Seventeen patients had severe underlying disease and 12 patients had mild or no underlying disease. Shock was present in 25 patients. Overall mortality was 62.1%. TNF-alpha was detected in nine patients (range: 57.7-3,169 pg/ml). There was a tendency to detect TNF-alpha in patients with mild or no underlying disease (p = 0.07). Detection of TNF-alpha was associated with survival (p = 0.0003) even when adjusted for severity of underlying disease (p = 0.005), shock (p = 0.0005), coagulation abnormality (p = 0.002) and immunosuppressive therapy (p = 0.005), using a bivariate analysis. In this investigation, detection of circulating TNF-alpha was predictive of good outcome in septic patients, suggesting a role for this cytokine in host-defense against this kind of infection.


Serum Neuroinflammatory Disease-Induced Central Nervous System Proteins Predict Clinical Onset of Experimental Autoimmune Encephalomyelitis.
There is an urgent need in multiple sclerosis (MS) patients to develop biomarkers and laboratory tests to improve early diagnosis, predict clinical relapses, and optimize treatment responses. In healthy individuals, the transport of proteins across the blood-brain barrier (BBB) is tightly regulated, whereas, in MS, central nervous system (CNS) inflammation results in damage to neuronal tissues, disruption of BBB integrity, and potential release of neuroinflammatory disease-induced CNS proteins (NDICPs) into CSF and serum. Therefore, changes in serum NDICP abundance could serve as biomarkers of MS. Here, we sought to determine if changes in serum NDICPs are detectable prior to clinical onset of experimental autoimmune encephalomyelitis (EAE) and, therefore, enable prediction of disease onset. Importantly, we show in longitudinal serum specimens from individual mice with EAE that pre-onset expression waves of synapsin-2, glutamine synthetase, enolase-2, and synaptotagmin-1 enable the prediction of clinical disease with high sensitivity and specificity. Moreover, we observed differences in serum NDICPs between active and passive immunization in EAE, suggesting hitherto not appreciated differences for disease induction mechanisms. Our studies provide the first evidence for enabling the prediction of clinical disease using serum NDICPs. The results provide proof-of-concept for the development of high-confidence serum NDICP expression waves and protein biomarker candidates for MS.


Vitamin D and its role in allergic disease.
In Western countries, the incidence of atopy and allergic diseases is high and further rising. While genetic factors certainly play a role, epigenetic or even nutritional factors might also be important in the pathogenesis of allergies. Vitamin D - the 'sunshine hormone' - exerts profound effects on both adaptive and innate immune functions involved in the development and course of allergic diseases. As also the incidence of vitamin D insufficiency is surprisingly high in the general population, clinical and experimental studies have started to investigate if correcting vitamin D levels [measured as serum 25 hydroxy vitamin D -25(OH)D] is beneficial or even protective in patients with allergies or children at risk. This review highlights current data on the effects of vitamin D on the allergy-mediating immune system and the vitamin D status in atopic patients. Furthermore, the benefits and risks of vitamin D supplementation during pregnancy, childhood and in adults with respect to the development and course of allergic disease are discussed.


Giant left pheochromocytoma with vascular anomalies and pelvic horseshoe kidney: a case report.
Pheochromocytoma is a neuroendocrine tumor, and its treatment is dependent on surgical resection. Due to the wide availability of cross-sectional imaging, pheochromocytomas are commonly seen as small tumors less than 10 cm in size and are mostly treated with minimally invasive surgery. Their concomitant presence with horseshoe kidney or other anatomical and vascular anomalies is rare. Herein, we present a surgically complex giant pheochromocytoma case who underwent an open left radical adrenalectomy. A 41-year-old Hispanic female presented with a 12 × 8 cm left hypervascular adrenal mass, pelvic horseshoe kidney, and severely dilated large left retro-aortic renal vein which branched into a left adrenal vein, congested left ovarian vein, and left uterine plexus. She was managed with insulin and metformin for uncontrolled diabetes with an A1c level of 9% and doxazosin for persistent hypertension. Clinical diagnosis of pheochromocytoma was confirmed with elevated urine and serum metanephrine and normetanephrine. The pre-operative ACTH was within normal range with a normal dexamethasone suppression test and 24-hour urine free cortisol. The adrenalectomy of the highly aggressive adrenal mass was performed via open approach to obtain adequate surgical exposure. Due to the large size of the tumor and its significant involvement with multiple adjacent structures, coordination with multiple surgical teams and close hemodynamic monitoring by anesthesiology was required for successful patient outcomes including preservation of blood supply to the pelvic horseshoe kidney. The histopathological diagnosis was pheochromocytoma with negative surgical margins. The patient was followed at 1, 4, 12, and 24 weeks postoperatively. She had a normal postoperative eGFR and was able to discontinue antihypertensive and antidiabetic medications at four weeks. She had transient adrenal insufficiency, which resolved at five months. The horseshoe kidney was intact except for a minimal area of hypo-enhancement in the left superior renal moiety due to infarction, which was significantly improved at six months. Our patient had a giant pheochromocytoma with anatomical variations complicating an already surgically challenging procedure. Nonetheless, with multiple provider collaboration, detailed pre-operative surgical planning, and meticulous perioperative monitoring, radical resection of the giant pheochromocytoma was safe and feasible with successful postoperative outcomes.


Evaluating the interactions between red stingray (Dasyatis akajei) melanocortin receptors and elephant shark (Callorhinchus milii) MRAP1 and MRAP2 following stimulation with either stingray ACTH(1-24) or stingray Des-Acetyl-αMSH: A pharmacological study in Chinese Hamster Ovary cells.
Previous studies on bony vertebrate MC2R orthologs (i.e., ray finned fishes, amphibians, reptiles, birds, and mammals) have shown that these MC2R orthologs have an obligatory requirement for interaction with bony vertebrate MRAP1 orthologs to a) allow for the trafficking of the MC2R ortholog to the plasma membrane; and b) to allow activation by ACTH, but not by any MSH-sized ligand. In addition, previous studies have found that co-expression of teleost and mammalian MC4R orthologs with corresponding MRAP2 has positive effects on sensitivity to stimulation by αMSH or ACTH. MRAP1 and MRAP2 paralogs have been detected in the genome of a cartilaginous fish (elephant shark), yet two cartilaginous fish MC2R orthologs (elephant shark and red stingray) do not apparently require MRAP1 for trafficking to the plasma membrane when expressed in Chinese Hamster Ovary (CHO) cells, and both orthologs can be activated by either ACTH or MSH-sized ligands. This study was done to determine whether sensitivity to stimulation by ACTH(1-24) or Des-Acetyl-αMSH is affected when stingray (sr) MC1R, MC2R, MC3R, MC4R or MC5R were co-expressed in CHO cells with either elephant shark (es) MRAP1 or esMRAP2. The results indicated that co-expression with heterologous MRAP1 increased the sensitivity of all five stingray melanocortin receptors for srACTH(1-24), but had not statistically significant effect on stimulation by srDes-Acetyl-αMSH for any of the stingray melanocortin receptors. Conversely, co-expression with esMRAP2 only enhanced sensitivity for srDes-Acetyl-αMSH for srMC4R, but had no effect on the other stingray orthologs, and there was no increase in sensitivity for srACTH(1-24) for any of the stingray melanocortin receptors. It appears then that some stingray melanocortin receptors have retained the ability to interact with a cartilaginous MRAP1 paralog. These results are discussed with reference to radiation of MRAP-related accessory proteins in cartilaginous fishes.


Evanescent Wave Optical Fiber Sensors Using Enzymatic Hydrolysis on Nanostructured Polyaniline for Detection of β-Lactam Antibiotics in Food and Environment.
β-Lactam antibiotics such as penicillins and cephalosporins are extensively used for human infection therapy. Consistent unintended exposure to these antibiotics via food and water is known to promote antibiotic-resistant bacterial pathogenesis with high morbidity and mortality in humans. An optical enzymatic biosensor for rapid and point-of-use detection of these antibiotics in food and water has been developed and tested. Enzymatic hydrolysis of β-lactams, on the electroactive polyaniline nanofibers, altered the polymeric backbone of the nanofibers, from emeraldine base form to emeraldine salt, which was measured as an increase in evanescent wave absorbance at 435 nm. The sensors were calibrated by spiking antibiotic-free milk with ceftazidime (as a model β-lactam analyte) in a linear range of 0.36-3600 nM (R2 = 0.98). The calibration was further validated for packaged milk, local cow milk, and buffalo milk. A similar calibration was devised for chicken meat samples in a linear range of 9-1800 nM (R2 = 0.982) and tap water in a linear range of 0.18-180 nM (R2 = 0.99). Interestingly, it was possible to use the same calibration for the determination of other β-lactam antibiotics (ampicillin, amoxicillin, and cefotaxime), which reflects the usefulness of the sensor for wide-scale deployment. The sensor performance was validated with a wastewater sample, from a wastewater treatment plant (WWTP), qualitatively analyzed by high-resolution liquid chromatography coupled with mass spectroscopy for detection of β-lactams. The sensor scheme developed and tested is of grassroot relevance as a quick solution for measurement of β-lactam residues in food and environment.


Assessing the International Transferability of a Machine Learning Model for Detecting Medication Error in the General Internal Medicine Clinic: Multicenter Preliminary Validation Study.
Although most current medication error prevention systems are rule-based, these systems may result in alert fatigue because of poor accuracy. Previously, we had developed a machine learning (ML) model based on Taiwan's local databases (TLD) to address this issue. However, the international transferability of this model is unclear. This study examines the international transferability of a machine learning model for detecting medication errors and whether the federated learning approach could further improve the accuracy of the model. The study cohort included 667,572 outpatient prescriptions from 2 large US academic medical centers. Our ML model was applied to build the original model (O model), the local model (L model), and the hybrid model (H model). The O model was built using the data of 1.34 billion outpatient prescriptions from TLD. A validation set with 8.98% (60,000/667,572) of the prescriptions was first randomly sampled, and the remaining 91.02% (607,572/667,572) of the prescriptions served as the local training set for the L model. With a federated learning approach, the H model used the association values with a higher frequency of co-occurrence among the O and L models. A testing set with 600 prescriptions was classified as substantiated and unsubstantiated by 2 independent physician reviewers and was then used to assess model performance. The interrater agreement was significant in terms of classifying prescriptions as substantiated and unsubstantiated (κ=0.91; 95% CI 0.88 to 0.95). With thresholds ranging from 0.5 to 1.5, the alert accuracy ranged from 75%-78% for the O model, 76%-78% for the L model, and 79%-85% for the H model. Our ML model has good international transferability among US hospital data. Using the federated learning approach with local hospital data could further improve the accuracy of the model.


Myocardial ischemia-reperfusion injury: Possible role of melatonin.
Our knowledge and understanding of the pathophysiology of coronary atherosclerosis has increased enormously over the last 20 years. Reperfusion through thrombolysis or percutaneous coronary angioplasty is the standard treatment for preventing acute myocardial infarction. Early reperfusion is an absolute prerequisite for survival of the ischemic myocardium, but reperfusion itself may lead to accelerated and additional myocardial injury beyond that generated by ischemia alone. These outcomes, in a range of reperfusion-associated pathologies, are collectively termed "reperfusion injuries". Reactive oxygen species are known to be produced in large quantities in the first few minutes of the post-ischemia reperfusion process. Similarly, scientific evidence from the last 15 years has suggested that melatonin has beneficial effects on the cardiovascular system. The presence of vascular melatoninergic receptor binding sites has been demonstrated; these receptors are functionally linked to vasoconstrictor or vasodilatory effects of melatonin. It has been shown that patients with coronary heart disease have a low melatonin production rate, especially those with higher risk of cardiac infarction and/or sudden death. Melatonin attenuates molecular and cellular damage resulting from cardiac ischemia-reperfusion in which destructive free radicals are involved.


[Epidemiological genetics of obesity in Northeast Mexico. Ascertainment of nuclear informative families].
The aims of this descriptive study in nuclear families were: 1) assess the distribution of nutritional stage of parents with respect to the nutritional stage of their children and 2) analyze those parents with the highest frequency of children at risk of being overweight (RS) and overweight (S). Body mass index (BMI) was determined in a sample of 126 nuclear families. Kappa values for parents and children were 0.95 and 0.97, respectively. For parents, birthplace of four grandparents was known. A wide variability in the nutritional stage (assessed by BMI) of the 252 parents (Normal-Normal, Normal-Obese, etc.) and their 300 children (Undernourished, Normal, RS and S) was found. A minimum number of children with RS and S (7.7%) was found among parents with Normal-Normal nutritional stage (reference group) in comparison with children whose parents were Overweight-Overweight (22%) and Overweight-Obese (35%). Since we found in parents Overweight-Overweight and Overweight-Obese a greater proportion of children with RS and S, this finding will facilitate the search for susceptibility genes for obesity in Mexican nuclear families with these characteristics.


["The intensive care simulator": a new teaching-concept to train severe sepsis management].
In addition to basic research and development of new therapeutic strategies, the education of health care professionals who manage sepsis patients is an important step to decrease the high mortality of severe sepsis. Patient simulators are increasingly used for teaching in anaesthesia. A training program in sepsis management was developed, using a full-scale anaesthesia simulator including the setting of a modern intensive care unit, and its results were evaluated by means of a questionnaire. The simulator is controlled from a separate room using a controlling computer provided with physiological models and pharmacokinetic as well as pharmacodynamic patterns of substances commonly used in anaesthesia and intensive care. An important element of the training program is the subsequent debriefing with different modules, according to the individual deficits and needs of the participants detected during simulation. From September 2002 to July 2004 82 physicians participated in the training program. 4 weeks after the training 52 % of the participants stated that they had changed their treatment behaviour due to the training content. They assessed the interactive simulator workshop semiquantitatively on a scale from 1 ("absolutely correct") to 7 ("not correct at all") as follows: Sepsis simulation training (SST) improves identification (mean+/-SD) (2.3 +/- 1.3) and treatment (2.5 +/- 1.2) of patients with severe sepsis, and SST including true-life scenarios is more appropriate than traditional lectures (1.5 +/- 0.7). The presented SST could be an effective way to train intensive care specialists in severe sepsis management.


Stimulation of appetite by ghrelin is regulated by leptin restraint: peripheral and central sites of action.
A reciprocal rhythmic pattern of 2 afferent hormonal signals, anorexigenic leptin and orexigenic ghrelin, imparts rhythmicity to the neuropeptide Y (NPY) system, the final common pathway for appetite expression in the hypothalamus. We now show that leptin inhibits both the secretion of gastric ghrelin and the stimulation of feeding by ghrelin. We propose that this dual leptin restraint is the major regulatory arm of the feedback communication between the periphery and the hypothalamus for weight homeostasis, and disruption in the rhythmic communication at any locus in the leptin-ghrelin-NPY feedback loop impels loss of hypothalamic control, leading to abnormal weight gain and obesity.


Repair of a complete anterior cruciate tear using prolotherapy: a case report.
OBJECTIVE AND IMPORTANCE: Surgical reconstruction is considered definitive treatment for anterior cruciate ligament (ACL) tears but precise surgical indications are debated. Some patients are reluctant or inappropriate surgical candidates. Prolotherapy is a non-surgical injection therapy for chronic musculoskeletal pain and instability. This case report documents the non-surgical repair of a torn ACL using prolotherapy and at-home exercise. CLINICAL PRESENTATION AND INTERVENTION: The 18 year old female patient sustained a right knee injury during a downhill skiing accident. Magnetic Resonance Imaging (MRI) revealed a high-grade partial versus complete rupture; Lachman exam findings suggested a complete rupture. She deferred surgical treatment. At 21 weeks post-injury, with unstable gait, inability to climb stairs and more than 1 cm anterior drawer test, she consented to undergo prolotherapy injections. She received 7 prolotherapy sessions over a 15 week period. At-home exercises were initiated at the 3(rd) prolotherapy session. RESULTS: The patient improved. Walking on flat ground improved 4 weeks after initiation of prolotherapy; she could ride a stationary bicycle for 30 minutes by 12 weeks. By 15 weeks, the patient had no instability climbing and descending stairs, the anterior drawer test was negative and MRI showed an intact ACL with fibrosis. Subsequently, she returned to full sport activity. CONCLUSIONS: We document the non-surgical repair of a high-grade partial or complete ACL tear using prolotherapy and at-home exercise. Prolotherapy may be an alternative to surgery in carefully selected patients. This report is consistent with findings of recent pilot-level studies and suggests the need for rigorous clinical trials assessing prolotherapy as treatment for ligament and tendon injury in selected patients.


Modelling the binding affinity of steroids to zebrafish sex hormone-binding globulin.
The circulating endogenous steroids are transported in the bloodstream. These are bound to a highly specific sex hormone-binding globulin (SHBG) and in lower affinity to proteins such as the corticosteroid-binding protein and albumin in vertebrates, including fish. It is generally believed that the glycoprotein SHBG protects these steroids from rapid metabolic degradation and thus intervenes in its availability at the target tissues. Endocrine disrupters binding to SHBG affect the normal activity of natural steroids. Since xenobiotics are primarily released in the aquatic environment, there is a need to evaluate the binding affinity of xenosteroid mimics on fish SHBG, especially in zebrafish (Danio rerio), a small freshwater fish originating in India and widely employed in ecotoxicology, toxicology, and genetics. In this context, a zebrafish SHBG (zfSHBG) homology model was developed using the human SHBG (hSHBG) receptor structure as template. It was shown that interactions with amino acids Ser-36, Asp-59 and Thr-54 were important for binding affinity. A ligand-based pharmacophore model was also developed for both zfSHBG and hSHBG inhibitors that differentiated binders from non-binders, but also demonstrated structural requirements for zfSHBG and hSHBG ligands. The study provides insights into the mechanism of action of endocrine disruptors in zebrafish as well as providing a useful tool for identifying anthropogenic compounds inhibiting zfSHBG.


Contralateral suppression of transient evoked otoacoustic emissions in children with phonological disorder.
Perception of acoustic details in the speech signal is important for speech sound development. The medial olivocochlear pathway, a part of the auditory efferent system, plays a role in stimulus-related control of the cochlea. One clinical tool to evaluate the medial olivocochlear activity, which is thought to improve speech perception in noise, is the suppression of otoacoustic emissions. This study investigated the suppression of transient evoked otoacoustic emissions in children with phonological disorder in comparison with that in typically developing controls. Case-control study. A total of 23 children with phonological disorder (aged 5-10 years) and 21 age- and sex-matched controls (P > 0.05) participated in the study. Participants had pure-tone thresholds ≤ 15 dB hearing loss and normal middle ear functions. Transient evoked otoacoustic emissions with and without contralateral acoustic stimulation were measured. Although the mean transient evoked otoacoustic emissions suppressions were lower in the group with phonological disorder than in the controls, these differences were not statistically significant (P > 0.05). No left/right ear asymmetry of transient evoked otoacoustic emissions suppression was detected in either of the groups (P > 0.05). Children with phonological disorder did not show alterations in medial olivocochlear functioning in the medial olivocochlear activity as measured by the contralateral suppression of transient evoked otoacoustic emissions.


Emperipolesis mediated by CD8 T cells is a characteristic histopathologic feature of autoimmune hepatitis.
Emperipolesis has been widely described in patients with autoimmune hepatitis, but the significance and the diagnostic value have not been quantitated. The goal of this study was to define the features and clinical significance of emperipolesis in autoimmune hepatitis (AIH). A retrospective histological evaluation of 101 patients with AIH and 184 controls was performed. Confocal staining for CD4, CD8, CD19, CD56, CD163, and CD11b, CK8/18 and cleaved caspase-3 was performed. Emperipolesis was observed in 65.3 % of the patients with AIH in haematoxylin and eosin (H&E)-stained slides, which was significantly higher than in patients with primary biliary cirrhosis (17.9 %), chronic hepatitis B (14.9 %), and drug-induced liver injury (25.6 %). Among AIH patients, the patients with emperipolesis had significantly higher serum (alanine aminotransferase/aspartate aminotransferase [ALT/AST]) levels. Histologically, emperipolesis was associated with more severe necroinflammatory features and more advanced fibrosis. The lymphocytes in hepatocytes were predominantly as CD8 T cells. Emperipolesis of CD8 T cells induced cleaved caspase-3 expression, and was prominent in areas apoptosis. Emperipolesis is a characteristic feature of AIH which is often seen in conjunction with interface hepatitis, plasmacytic infiltration and hepatocyte rosetting and is associated with more severe necroinflammatory and fibrotic changes. In AIH, emperipolesis is predominantly mediated by CD8 T cells, appears to induce apoptosis and may be another mechanism of autoimmune-mediated hepatocyte injury.


Further validation of a model of fibromyalgia syndrome in the rat.
We have recently developed an animal model of fibromyalgia syndrome in the rat. In this model, rats exposed to unpredictable sound stress develop a delayed onset enhancement and prolongation of cytokine-induced mechanical hyperalgesia in muscle and skin. In this study, we tested the hypothesis that our model also manifests symptoms of common comorbid diagnoses: irritable bowel syndrome, temporomandibular disorder, and anxiety. Both visceral sensitivity and cytokine hyperalgesia in masseter muscle were present in the stressed rats. Furthermore, in an established model of irritable bowel syndrome-water avoidance-we observed significant muscle hyperalgesia. Finally, using the elevated plus maze to assess for anxiety level, we observed a significantly higher anxiety level in sound stress-exposed rats. Thus, unpredictable sound stress produces a condition in the rat with several features-delayed onset visceral and temporomandibular hyperalgesia and increased anxiety, as well as cutaneous and muscle hyperalgesia-commonly found in patients with fibromyalgia syndrome. A stress model-unpredictable sound-in the rat exhibits several features (cutaneous, musculoskeletal, and visceral hyperalgesia, as well as anxiety) that are found in patients with fibromyalgia syndrome. Thus, this model may be used to test hypotheses about the underlying mechanisms and response to therapy in patients with fibromyalgia.


Light-controlled nanoswitches: from fabrication to photoelectric switching.
Plasmon-assisted optical growth significantly improves the controllability of polymerization on the nanoscale, which greatly facilitates the fabrication of nanodevices and their integration. Traditional approaches for nanodevices strongly rely on nanofabrication techniques, which involve complicated multi-steps and expensive instrumentation. Here, we build photoelectric nanoswitches by precisely implanting photoelectric active polymers, polypyrrole, into metallic bowtie nanogaps via charge transfer induced oxidative polymerization, which show conductivity up to 4 μS m-1 with an optical switching response time of ∼50 ms. We further demonstrate their photoelectric switchability with different laser powers, wavelengths and amounts of polypyrrole in the gaps. This plasmon-assisted optical growth strategy not only provides a facile and accurate means to control the polymerization process, but also greatly expedites the fabrication of nanodevices and their integration.


Long-term and realistic global change manipulations had low impact on diversity of soil biota in temperate heathland.
In a dry heathland ecosystem we manipulated temperature (warming), precipitation (drought) and atmospheric concentration of CO2 in a full-factorial experiment in order to investigate changes in below-ground biodiversity as a result of future climate change. We investigated the responses in community diversity of nematodes, enchytraeids, collembolans and oribatid mites at two and eight years of manipulations. We used a structural equation modelling (SEM) approach analyzing the three manipulations, soil moisture and temperature, and seven soil biological and chemical variables. The analysis revealed a persistent and positive effect of elevated CO2 on litter C:N ratio. After two years of treatment, the fungi to bacteria ratio was increased by warming, and the diversities within oribatid mites, collembolans and nematode groups were all affected by elevated CO2 mediated through increased litter C:N ratio. After eight years of treatment, however, the CO2-increased litter C:N ratio did not influence the diversity in any of the four fauna groups. The number of significant correlations between treatments, food source quality, and soil biota diversities was reduced from six to three after two and eight years, respectively. These results suggest a remarkable resilience within the soil biota against global climate change treatments in the long term.


The bone compartment in a teleost fish, Ictalurus punctatus: size, composition and acid-base response to hypercapnia.
An estimate of the total mass of bone in the Channel catfish Ictalurus punctatus Rafinesque, was obtained by dissection. The wet weight of bone constituted 16.3 +/- 1.9% (+/- S.D.) of the total (live) wet weight, and 25.0 +/- 2.1% of the dry weight. Of the dry skeletal material, 66.3 +/- 11.1% was soluble in strong acid. The acid-soluble material was about half mineral salts, consisting of 19.5 +/- 2.21% Ca2+ and 27.6 +/- 3.22% PO4(3-), with minor fractions of Mg2+ (0.33%) and CO3(2-) (1.67%). The pH values of fluid compartments associated with skull and vertebral bone tissues were 7.420 +/- 0.026 and 7.444 +/- 0.017 (+/- S.E.), respectively, at a normocapnic plasma pH of 7.868 +/- 0.020. In response to external hypercapnia (7.5 Torr), the blood response consisted of an immediate decrease in pH, and a subsequent compensatory rise in both pH and [HCO3-]. This compensatory phase was accompanied by a net apparent H+ excretion to the water. The participation of the mineral salts of the bone compartment in compensation appeared to be negligible, since there was no significant change in either blood [Ca2+] or [PO4(3-)], nor any significant increase in calcium efflux to the water. The intracellular pH values of the bone compartments were only slightly higher than other tissues, and the changes in pHi during hypercapnia were similar in bone and white muscle. Thus, the bone compartment in the fish appears to be well regulated, relatively refractory to acute acid-base disturbance, and does not serve as an ion source during acid-base compensation.


Variable Surface Glycoprotein from Trypanosoma brucei Undergoes Cleavage by Matrix Metalloproteinases: An in silico Approach.
In order to survive as extracellular parasites in the mammalian host environment, Trypanosoma brucei has developed efficient mechanisms of immune system evasion, which include the abundant expression of a variable surface glycoprotein (VSG) coat. VSGs are anchored in the parasite membrane by covalent C-terminal binding to glycosylphosphatidylinositol and may be periodically removed by a phospholipase C (PLC) and a major surface protein (TbMSP). VSG molecules show extraordinary antigenic diversity and a comparative analysis of protein sequences suggests that conserved elements may be a suitable target against African trypanosomiasis. However, the cleavage mechanisms of these molecules remain unclear. Moreover, in protozoan infections, including those caused by Trypanosoma brucei, it is possible to observe an increased expression of the matrix metalloproteinases (MMPs). To address the cleavage mechanism of VSGs, the PROSPER server was used for the identification of VSG sequence cleavage sites. After data compilation, it was observed that 64 VSG consensus sequences showed a high conservation of hydrophobic residues, such as valine (V), methionine (M), leucine (L) and isoleucine (I) in the fifth position-the exact location of the cleavage site. In addition, the PROSPER server identified conserved cleavage site portions of VSG proteins recognized by three matrix metalloproteases (gelatinases: MMP-2, MMP-3 and MMP-9). However, further biological studies are needed in order to analyze and confirm this prediction.


Gustatory perception and metabolic utilization of sugars by Myrmica rubra ant workers.
The suitability of various nectar and honeydew sugars as a food source for the polyphagous ant species M. rubra (L.) was studied. The sugars used included monosaccharides (fructose, glucose, galactose, mannose, rhamnose), disaccharides (sucrose, maltose, trehalose, melibiose, lactose) and trisaccharides (melizitose, raffinose, erlose). Single-sugar solutions were tested on ant workers in a long-term laboratory bioassay in which acceptance of the solutions and ant survival were recorded. The acceptance of the sugars was confirmed in a second bioassay in which feeding time was established. Enzymatic hydrolysis of sucrose, maltose and melibiose was investigated through HPLC analyses of workers fed these disaccharides. Sugar acceptance and feeding time were related to ant survival. Considering the monosaccharide units of which the sugars are composed, fructose seems especially suitable as a short-term energy source, while glucose appears to be used both directly and for storage. The presence of a galactose unit appears to reduce sugar suitability. It is suggested that the workers possess invertase and maltase and to a lesser degree also galactosidase. The gustatory perception is correlated with the profitability of sugars in further metabolic processes.


Single-Cell Analysis of Human Pancreas Reveals Transcriptional Signatures of Aging and Somatic Mutation Patterns.
As organisms age, cells accumulate genetic and epigenetic errors that eventually lead to impaired organ function or catastrophic transformation such as cancer. Because aging reflects a stochastic process of increasing disorder, cells in an organ will be individually affected in different ways, thus rendering bulk analyses of postmitotic adult cells difficult to interpret. Here, we directly measure the effects of aging in human tissue by performing single-cell transcriptome analysis of 2,544 human pancreas cells from eight donors spanning six decades of life. We find that islet endocrine cells from older donors display increased levels of transcriptional noise and potential fate drift. By determining the mutational history of individual cells, we uncover a novel mutational signature in healthy aging endocrine cells. Our results demonstrate the feasibility of using single-cell RNA sequencing (RNA-seq) data from primary cells to derive insights into genetic and transcriptional processes that operate on aging human tissue.


DGaze: CNN-Based Gaze Prediction in Dynamic Scenes.
We conduct novel analyses of users' gaze behaviors in dynamic virtual scenes and, based on our analyses, we present a novel CNN-based model called DGaze for gaze prediction in HMD-based applications. We first collect 43 users' eye tracking data in 5 dynamic scenes under free-viewing conditions. Next, we perform statistical analysis of our data and observe that dynamic object positions, head rotation velocities, and salient regions are correlated with users' gaze positions. Based on our analysis, we present a CNN-based model (DGaze) that combines object position sequence, head velocity sequence, and saliency features to predict users' gaze positions. Our model can be applied to predict not only realtime gaze positions but also gaze positions in the near future and can achieve better performance than prior method. In terms of realtime prediction, DGaze achieves a 22.0% improvement over prior method in dynamic scenes and obtains an improvement of 9.5% in static scenes, based on using the angular distance as the evaluation metric. We also propose a variant of our model called DGaze_ET that can be used to predict future gaze positions with higher precision by combining accurate past gaze data gathered using an eye tracker. We further analyze our CNN architecture and verify the effectiveness of each component in our model. We apply DGaze to gaze-contingent rendering and a game, and also present the evaluation results from a user study.


Human papillomavirus infection and vaccination: Knowledge and attitudes among young males in Italy.
This study assessed knowledge and attitudes about Human papillomavirus (HPV) and the relative vaccination and their determinants in a sample of young males. The survey was conducted between January and April 2015 among a sample of 1000 males aged between 14-24 y in the geographic area of Naples and Caserta, Italy. The 54.9% of the participants reported of having heard about the HPV infection. Those who were aware about the availability of the vaccine, who reported the first vaginal sexual encounter before the 18 y and at least at 18 y compared to those who had not had a complete sexual intercourse, who had undergone a health checkup in the last year, and who had received information about the HPV vaccine by physicians had a significant higher knowledge about the HPV infection. The 58.2% reported that they would be willing to receive the HPV vaccine. Those younger, who reported the first vaginal sexual encounter at least at 18 y, who agreed that male should receive the vaccine, who knew that both males and females can acquire the infection, and who agreed that the vaccine is an important preventive intervention, expressed more positive attitude toward willingness to receive the vaccine. More information about the HPV vaccine were required by those who agreed that the vaccine is an important preventive intervention, who reported the first vaginal sexual encounter at least at 18 y, who have had only one partner in the last year compared to students who had no partner, and who had received information about the vaccine by physicians. This study highlights a need for improved education of young males of the HPV infection and the associated diseases and about the benefit of the vaccination.


Effects of medulloblastoma resections on outcome in children: a report from the Children's Cancer Group.
We reviewed the data of children with high-stage primitive neuroectodermal tumors (medulloblastomas) who were treated on Children's Cancer Group-921 protocol to evaluate the correlation between tumor resection and prognosis. Patients enrolled in the study had either tumors that were operatively categorized to be Chang tumor stage 3b or 4, postoperative residual tumors > 1.5 cm2, or evidence of tumor dissemination (Chang metastasis Stages [M Stages] 1-4) at diagnosis. Resections were analyzed in two ways, as follows: 1) by the extent of resection (percent of the tumor that was removed), as estimated by the treating neurosurgeon; and 2) by the extent of residual tumor (how much of the tumor was left), as estimated from postoperative scans. Two hundred and three children were enrolled in the study with institutional diagnoses of primitive neuroectodermal tumors-medulloblastomas; diagnoses were confirmed by central neuropathological review in 188 patients. Progression-free survival (PFS) at 5 years was 54% (standard error, 5%). As in previous Children's Cancer Group studies, age and M stage correlated with survival; PFS was significantly lower in children 1.5 to 3.0 years old at diagnosis and in those with any evidence of tumor dissemination (M Stage 1-4). On univariate analysis, neither extent of resection nor extent of residual tumor correlated with PFS. However, adjusting for other factors, extent of residual tumor was important; PFS was 20% (standard error, 14%) better at 5 years in children with no dissemination (M Stage 0) who had < 1.5 cm2 of residual tumor (P = 0.065) and was 24% (standard error, 14%) better at 5 years in children > 3 years old with no tumor dissemination (M Stage 0) and with < 1.5 cm2 residual tumor (P = 0.033). On the basis of our observations, we conclude that extent of tumor resection, as estimated by the neurosurgeon, does not correlate with outcome but that extent of residual tumor does correlate with prognosis in certain children (those who are > 3 years old, with no tumor dissemination). In contrast to age and M stage, the major factors associated with outcome, residual tumor is an important variable in outcome, one that neurosurgeons can control.


Hypothalamic and myocardial lesions after subarachnoid haemorrhage.
The hypothalamus and myocardium of 12 patients who had died after a subarachnoid haemorrhage, and of six patients who had died from other intracranial pathology were examined. Only in the patients who had died from subarachnoid haemorrhage were histological lesions found in both the hypothalamus and myocardium. The possible significance of these findings is discussed with particular reference to the sympathetic nervous system.


Leukocyte telomere length independently predicts hyperuricemia risk in a longitudinal study of the Chinese population.
Leukocyte telomere length (LTL) has been correlated with uric acid levels, although results are inconsistent, and prospective studies are lacking. In this longitudinal, prospective cohort study, we aimed to assess whether a shorter LTL predicts the risk of hyperuricemia. We conducted a longitudinal study in a Chinese cohort of 599 participants. Of these, 266 participants completed a 5.9-year follow-up from June 2014 to December 2021. LTL was assessed at baseline using real-time polymerase chain reaction. Hyperuricemia was defined as serum uric acid ≥420 mmol/L according to Chinese guidelines for diagnosis and treatment of hyperuricemia and gout. Participants who had developed hyperuricemia during follow-up (n = 17) had shorter LTL at baseline. Baseline LTL was independently associated with the development of hyperuricemia at follow-up after adjusting for conventional hyperuricemia risk factors (odds ratio [OR] 2.347 [95% confidence interval [CI] 1.123, 4.906]; P = 0.023). After grouping according to LTL tertiles, the incidence of hyperuricemia was 18.334-fold higher for the first than for the third tertile (OR 18.334 [95%CI 1.786, 191.272]; P = 0.014, P for trend = 0.050). Our findings in a prospective cohort suggest that LTL could predict hyperuricemia risk, which might inform the timely prevention and treatment of hyperuricemia.


Crocin-Elicited Autophagy Rescues Myocardial Ischemia/Reperfusion Injury via Paradoxical Mechanisms.
Crocin, the main effective component of saffron, exerts protective effects against ischemia/reperfusion injury during strokes. However, the effects of crocin in myocardial ischemia/reperfusion injury, and the mechanisms involved, remain unknown. Pretreated with crocin for 7 days, C57BL/6N mice were subjected to 30 min of myocardial ischemia followed by 12[Formula: see text]h of reperfusion (for cardiac function and infarct size, cell apoptosis and necrosis). Neonatal mouse cardiomyocytes were subjected to 2 h of hypoxia followed by 4 h of reoxygenation. NMCM's survival was assessed during hypoxia and reoxygenation in the presence or absence of the autophagy inhibitor 3-methyladenine or the inducer rapamycin. Western blotting was used to evaluate AMPK, Akt, and autophagy-related proteins. Autophagosome was observed using electron microscopy. In the in vivo experiment, crocin pretreatment significantly attenuated infarct size, myocardial apoptosis and necrosis, and improved left ventricular function following ischemia/reperfusion. In vitro data revealed that autophagy was induced during hypoxia, the levels of which were intensely elevated during reoxygenation. Crocin significantly promoted autophagy during ischemia, accompanied with the activation of AMPK. In contrast, crocin overtly inhibited autophagy during reperfusion, accompanied with Akt activation. Induction and inhibition of autophagy mitigated crocin induced protection against NMCMs injury during hypoxia and reoxygenation, respectively. Our data suggest that crocin demonstrated a myocardial protective effect via AMPK/mTOR and Akt/mTOR regulated autophagy against ischemia and reperfusion injury, respectively.


Strong effects of amoebae grazing on the biomass and genetic structure of a Microcystis bloom (Cyanobacteria).
Despite its importance for bloom toxicity, the factors determining the population structure of cyanobacterial blooms are poorly understood. Here, we report the results of a two-year field survey of the population dynamics of Microcystis blooms in a small hypertrophic urban pond. Microscopic enumeration of Microcystis and its predators and parasites was combined with pigment and microcystin analysis and denaturing gradient gel electrophoresis of the ITS rDNA region to assess population dynamics and structure. Two main Microcystis morpho- and ITS types were revealed, corresponding to M. aeruginosa and M. viridis. In both years, high population densities of naked amoebae grazing on Microcystis coincided with rapid decreases in Microcystis biomass. In one year, there was a shift from heavily infested M. aeruginosa to the less-infested M. viridis, allowing the bloom to rapidly recover. The preference of amoebae for M. aeruginosa was confirmed by grazing experiments, in which several amoeba strains were capable of grazing down a strain of M. aeruginosa, but not of M. viridis. Zooplankton and chytrid parasites appeared to be of minor importance for these strong and fast reductions in Microcystis biomass. These findings demonstrate a strong impact of small protozoan grazers on the biomass and genetic structure of Microcystis blooms.


Trust in provider and stigma during second-trimester abortion.
To determine whether trust in the provider and sociodemographics are associated with individual-level abortion stigma. We performed a cross sectional and exploratory study design using secondary analysis of a randomized trial that enrolled participants undergoing second trimester abortion. We collected baseline survey data from 70 trial participants to assess stigma (Individual Level of Abortion Stigma scale, ILAS; range 0-4), trust in provider (Trust in Physician scale; range 1-5), anxiety, depression, and sociodemographics. We performed multiple linear regression, for which ILAS score was the outcome of interest. Univariate associations were used to inform the regression model. The mean abortion stigma score was at the low end of the ILAS at 1.21 (range 0.2-2.8, SD 0.66). Age, race, income, BMI, parity, gestational age at time of abortion, and reasons for ending the pregnancy were not significantly associated with the ILAS score. Higher trust in provider scores were (m 4.0, SD 0.49) and inversely related to the ILAS score, even after adjustment for confounders (β -0.02, CI -0.03 to -0.004, p = 0.013). Screening positive for anxiety or depression was associated with a higher ILAS score ((β 0.48, CI 0.10, 0.90, p = 0.015); (β = 0.27 CI -0.097, 0.643)), while cohabitation was associated with lower ILAS score (β -0.44, CI -0.82 to -0.57, p = 0.025). Trust in an abortion provider, anxiety, depression, and cohabitation are associated with abortion stigma among people seeking second trimester abortion care. Interventions that improve trust in a provider may be an area of focus for addressing abortion stigma. Future research should confirm these findings in larger populations and across diverse locations and demographics and to conduct qualitative research to understand what patients perceive as trust-promoting behaviors and words during abortion encounters.


Determination of MDA for a two count method for stripping short-lived activity out of an air sample.
When measuring airborne activity of isotopes with very low annual limit of intakes, interference from natural radioactivity presents an obstacle to obtaining timely results. One method for solving this problem is to perform two separate counts of the sample and mathematically strip-out the contribution from the short-lived component. While this method can process a large volume of samples with little or no preparation time, the sensitivity of the analysis is hard to determine. In an attempt to determine the sensitivity of the analysis and to quantify the parameters that affect it, an equation for the minimum detectable activity was derived. Actual air sample data were then evaluated against this equation to determine its usefulness.


The modulatory effect of ellagic acid and rosmarinic acid on ultraviolet-B-induced cytokine/chemokine gene expression in skin keratinocyte (HaCaT) cells.
Ultraviolet radiation (UV) induces an increase in multiple cutaneous inflammatory mediators. Ellagic acid (EA) and rosmarinic acid (RA) are natural anti-inflammatory and immunomodulatory compounds found in many plants, fruits, and nuts. We assessed the ability of EA and RA to modulate IL-1β, IL-6, IL-8, IL-10, MCP-1, and TNF-α gene expression in HaCaT cells after UVB irradiation. Cells were treated with UVB (100 mJ/cm(2)) and simultaneously with EA (5 μM in 0.1% DMSO) or RA (2.7 μM in 0.5% DMSO). Moreover, these substances were added to the UVB-irradiated cells 1 h or 6 h before harvesting, depending on the established UVB-induced cytokine expression peak. Cytokine gene expression was examined using quantitative real time polymerase chain reaction. RA produced a significant reduction in UVB-induced expression of IL-6, IL-8, MCP-1, and TNF-α when applied at the same time as irradiation. EA showed milder effects compared with RA, except for TNF-α. Both substances decreased IL-6 expression, also when applied 5 h after irradiation, and always produced a significant increase in UVB-induced IL-10 expression. Our findings suggest that EA and RA are able to prevent and/or limit the UVB-induced inflammatory cascade, through a reduction in proinflammatory mediators and the enhancement of IL-10, with its protective function.


Use of cellulose nanofibers as a denture immersing solution.
The purpose of this study was to investigate the influence of cellulose nanofibers (CNF) solution on the mechanical and biological properties of denture base resins (DBR). Two types of CNFs obtained from bamboo (BB) and needle-leaved (NB) trees were used in this study. We prepared 18 different CNF solutions based on their fibrillation (A-low, B-middle, and C-high) and concentration (0.05, 0.10, and 0.20 wt%). A heat-polymerized acrylic resin was used as DBR. The contact angles for each specimen were measured after immersion. The flexural properties of the immersed specimens, and the biological properties of the CNF solutions were examined. Specimens immersed in CNF-NB-C-0.05 wt% solution presented with the lowest contact angles. Specimens in CNF-NB-C and CNF-BB-A groups showed higher flexural modulus values. No cytotoxic or antimicrobial effects were observed for the CNF solutions. This study suggest that CNF solution may improve the surface wettability of the DBR without affecting its flexural property.