Tofacitinib (CP-690,550), a Janus kinase inhibitor for dry eye disease: results from a phase 1/2 trial.
To evaluate safety and efficacy of topical ophthalmic tofacitinib (CP-690,550), a novel Janus kinase inhibitor, in treating dry eye disease (DED). A phase 1/2 prospective, randomized, double-masked, multicenter, vehicle- and comparator-controlled trial (NCT00784719). Patients (n = 327) 18 years of age and older with a DED diagnosis for 6 months or more. Tofacitinib (0.0003% twice daily, n = 46; 0.001% in both eyes twice daily, n = 47; 0.003% twice daily, n = 48; 0.005% twice daily, n = 48; 0.005% once daily, n = 44) results were compared with those of groups receiving active treatment cyclosporine ophthalmic emulsion 0.05% twice daily (n = 47) and vehicle twice daily (n = 47). Safety and efficacy evaluations were performed at baseline and throughout the 8-week study. Schirmer wetting, corneal staining, tear film break-up time, conjunctival staining, Ocular Comfort Index (OCI), and Ocular Surface Disease Index (OSDI). All tofacitinib doses were well tolerated, exhibiting better patient-reported ocular tolerability than cyclosporine. For the proportion of patients achieving 10 mm or more Schirmer wetting (without anesthesia) at week 8 (primary end point), greater response rates were observed in the tofacitinib 0.001% twice daily (27.3%), 0.005% twice daily (25.5%), and 0.005% once daily (26.1%) groups versus vehicle (20.0%); however, the differences were not statistically significant. Mean increase in Schirmer wetting (without anesthesia) from baseline was statistically significant (P<0.2, 2-sided) for all tofacitinib doses (1.7-3.1 mm), cyclosporine (3.9 mm), and vehicle (1.4 mm). For corneal staining (total score), significant improvement (reduction) from baseline was observed for all tofacitinib doses (-0.9 to -1.9) and vehicle (-2.0), but not for cyclosporine. The proportion of patients with complete corneal clearing (CCC; 100%) at week 8 was greatest with tofacitinib 0.005% once daily (15.9%) versus vehicle (6.7%). Symptom scores (OCI, OSDI) at week 8 showed significant improvements from baseline for all tofacitinib groups, and tofacitinib demonstrated greater improvements than cyclosporine. The tofacitinib 0.005% once daily group showed significant improvements in both a sign (Schirmer wetting without anesthesia) and symptom (OSDI environmental triggers subscale) versus vehicle and also demonstrated the highest response rate for CCC (16.7%) at week 8. This phase 1/2 study of tofacitinib demonstrated a trend for improving both signs and symptoms of dry eye. All doses of tofacitinib exhibited a reasonable safety profile and were well tolerated by patients with DED.


Selective nonoperative management of blunt liver injury using computed tomography.
Computed tomographic (CT) scans are used to evaluate victims of blunt trauma for abdominal injury when reasons for immediate laparotomy are not present. Twenty-four patients whose CT scans showed liver injuries that were small parenchymal lacerations or intrahepatic hematomas were managed without laparotomy. Intra-abdominal blood was absent or estimated to be less than 250 mL in volume. None of the patients were in shock; six patients required transfusions, none for acute abdominal bleeding. None of these patients subsequently required laparotomy or showed hepatobiliary problems at a follow-up examination less than one year later. Five follow-up scans showed varying degrees of resolution. Two patients died of severe head injury, but the liver injury did not contribute to the cause of death. Other patients with more severe blunt liver injury who were treated nonoperatively developed significant complications that required delayed surgery. It seems that patients with limited liver injury diagnosed by CT scan and selected by strictly applied criteria can be managed safely without laparotomy in a setting where rapid evaluation and treatment of any potential complication is available.


A painful glomus tumor on the pulp of the distal phalanx.
A 52-year-old female patient presented with an 8-year history of progressively intense pain, cold sensitivity, and severe tenderness to palpation of the ulnar side of the tip of her right little finger. Subsequent diagnostic evaluation with ultrasonographic imaging revealed the presence of a glomus tumor in the tender area. Glomus tumors are benign, occurring in the vascular hamartomatous tubercles of the glomus body, which is a myoarterial apparatus typically found in the reticular dermis of the skin. Distal glomus tumors are relatively uncommon, and account for approximately 1% of all hand tumors. Most of them are located in the subungual area because of its high concentration of glomus bodies. We report a case of a glomus tumor with a typical triad of symptoms, yet with a rare location : on the pulp of the ulnar aspect of the distal phalanx of the right little finger.


Echium oil reduces plasma triglycerides by increasing intravascular lipolysis in apoB100-only low density lipoprotein (LDL) receptor knockout mice.
Echium oil (EO), which is enriched in SDA (18:4 n-3), reduces plasma triglyceride (TG) concentrations in humans and mice. We compared mechanisms by which EO and fish oil (FO) reduce plasma TG concentrations in mildly hypertriglyceridemic male apoB100-only LDLrKO mice. Mice were fed one of three atherogenic diets containing 0.2% cholesterol and palm oil (PO; 20%), EO (10% EO + 10% PO), or FO (10% FO + 10% PO). Livers from PO- and EO-fed mice had similar TG and cholesteryl ester (CE) content, which was significantly higher than in FO-fed mice. Plasma TG secretion was reduced in FO vs. EO-fed mice. Plasma very low density lipoprotein (VLDL) particle size was ordered: PO (63 ± 4 nm) > EO (55 ± 3 nm) > FO (40 ± 2 nm). Post-heparin lipolytic activity was similar among groups, but TG hydrolysis by purified lipoprotein lipase was significantly greater for EO and FO VLDL compared to PO VLDL. Removal of VLDL tracer from plasma was marginally faster in EO vs. PO fed mice. Our results suggest that EO reduces plasma TG primarily through increased intravascular lipolysis of TG and VLDL clearance. Finally, EO may substitute for FO to reduce plasma TG concentrations, but not hepatic steatosis in this mouse model.


Dissociation of the effects of epinephrine and insulin on glucose and protein metabolism.
The separate and combined effects of insulin and epinephrine on leucine metabolism were examined in healthy young volunteers. Subjects participated in four experimental protocols: 1) euglycemic insulin clamp (+80 microU/ml), 2) epinephrine infusion (50 ng.kg-1.min-1) plus somatostatin with basal replacement of insulin and glucagon, 3) combined epinephrine (50 ng.kg-1.min-1) plus insulin (+80 microU/ml) infusion, and 4) epinephrine and somatostatin as in study 2 plus basal amino acid replacement. Studies were performed with a prime-continuous infusion of [1-14C]leucine and indirect calorimetry. Our results indicate that 1) hyperinsulinemia causes a generalized decrease in plasma amino acid concentrations, including leucine; 2) the reduction in plasma leucine concentration is primarily due to an inhibition of endogenous leucine flux; nonoxidative leucine disposal decreases after insulin infusion; 3) epinephrine, without change in plasma insulin concentration, reduces plasma amino acid levels; 4) combined epinephrine-insulin infusion causes a greater decrease in plasma amino levels than observed with either hormone alone; this is because of a greater inhibition of endogenous leucine flux; and 5) when basal amino acid concentrations are maintained constant with a balanced amino acid infusion, epinephrine inhibits the endogenous leucine flux. In conclusion, the present results do not provide support for the concept that epinephrine is a catabolic hormone with respect to amino acid-protein metabolism. In contrast, epinephrine markedly inhibits insulin-mediated glucose metabolism.


Seven-up, the Drosophila homolog of the COUP-TF orphan receptors, controls cell proliferation in the insect kidney.
Morphogenesis of the insect kidney, the Malpighian tubules, is controlled in Drosophila by a single large cell, the tip cell. It has been postulated that this cell sends out a mitogenic signal that induces the division of neighboring cells. The signal and the molecules that receive it have remained elusive. We show that the COUP-TF-related nuclear orphan receptor Seven-up is a key component that becomes induced in response to mitogenic EGF receptor signaling activity emanating from the tip cell. Seven-up in turn is capable of regulating the transcription of cell cycle regulators.


Optical manipulation for studies of collisional dynamics of micron-sized droplets under gravity.
A new experimental technique for creating and imaging collisions of micron-sized droplets settling under gravity is presented. A pair of glycerol droplets is suspended in air by means of two optical traps. The droplet relative velocities are determined by the droplet sizes. The impact parameter is precisely controlled by positioning the droplets using the two optical traps. The droplets are released by turning off the trapping light using electro-optical modulators. The motion of the sedimenting droplets is then captured by two synchronized high-speed cameras, at a frame rate of up to 63 kHz. The method allows the direct imaging of the collision of droplets without the influence of the optical confinement imposed by the trapping force. The method will facilitate efficient studies of the microphysics of neutral, as well as charged, liquid droplets and their interactions with light, electric field and thermodynamic environment, such as temperature or vapor concentration.


Synthesis of the tumor associative α-aminooxy disaccharide of the TF antigen and its conjugation to a polysaccharide immune stimulant.
The α-aminooxy derivative of the Thomsen-Friedenriech tumor associated carbohydrate antigen has been synthesized in 11 steps utilizing a D-GalN3 acceptor carrying a pre-installed α-N-hydroxysuccinimidyl moiety. The natural α linkage was prepared in high selectivity employing a suitably protected D-GalN3-thioglycoside donor with N-hydroxysuccinimide. With access to α-TF-ONH2, the preparation of the TF-PS A1 vaccine candidate ensued smoothly through oxime bond formation.


Client attachment in a randomized clinical trial of psychoanalytic and cognitive-behavioral psychotherapy for bulimia nervosa: Outcome moderation and change.
In the context of a randomized clinical trial of psychoanalytic psychotherapy (PPT) versus cognitive behavior therapy (CBT) for bulimia nervosa (BN), this study performed secondary analyses of (a) the relation between attachment and pretreatment symptom levels, (b) whether client pretreatment attachment moderated treatment outcome, (c) whether change in client attachment was associated with symptomatic change, and (d) whether client attachment changed differently in the 2 treatments. Sixty-nine women and 1 man of a mean age of 25.8 years diagnosed with BN were randomly assigned to either 2 years of weekly PPT or 5 months of CBT. Assessments at intake, after 5 months, and after 2 years included the Eating Disorder Examination to assess eating disorder symptoms, the Adult Attachment Interview to assess client attachment, and the Symptom Checklist 90-R to assess general psychiatric distress. Repeated measures were analyzed using multilevel analysis. Higher scores on attachment insecurity and attachment preoccupation were associated with more frequent binging pretreatment. Pretreatment attachment did not predict treatment outcome. In PPT, but not in CBT, reduction of binging was associated with an increase in attachment security. The 2 treatment types were not associated with significantly different patterns of attachment-related change. Degree and type of attachment insecurity is related to the frequency of binging in BN. Increase in attachment security may be a treatment-specific mechanism of change in PPT for BN. (PsycINFO Database Record


Sentinel lymph node biopsy in early melanoma-comparison of two techniques for sentinel removal.
Sentinel lymph node biopsy is an established technique for melanoma staging. There are no studies available comparing different techniques for the removal of sentinel lymph nodes related to safety and postsurgical complication rate. This is a retrospective single-center trial. We analyzed the postsurgical complications in 201 consecutive melanoma patients with ligature of lymphatic vessels by sutures (group A) and in 91 consecutive patients with occlusion of lymphatic vessels by bipolar tweezers (group B). We paid particular attention to complications related to disturbed lymphatic function, such as lymph edema, lymphatic fistula, and seroma. The complication rate was 5.5 % (group A) and 9.6 % (group B) which is in the range of other published trials (p = 0.89). There was no increase of complications related to lymphatic vessels in group B, although the rate of patients with more than two sentinel lymph nodes removed was 5‑times higher than in group A. Removal of sentinel lymph nodes with the use of bipolar tweezers does not increase the risk of postsurgical complications, and in particular it is not associated with a higher rate of complications related to lymphatic vessel dysfunction.


Attention bias modification training for adolescents with chronic pain: a randomized placebo-controlled trial.
Attention bias for pain-related information is theorised to maintain chronic pain, indicating that changing this bias could improve pain-related outcomes. Modifying attention biases in adolescents, when chronic pain often first emerges, may be particularly beneficial. We report here a randomized, placebo-controlled, parallel-group trial of attention bias modification (ABM) training in adolescents with chronic noncancer pain. Adolescent patients (N = 66) were randomly assigned to complete multiple sessions of dot-probe ABM training (N = 23), placebo training (N = 22), or no training (waitlist; N = 21) across a period of 4 weeks. Patients completed all assessments at a hospital-based pediatric pain clinic and completed all training at home. We examined the relative effects of ABM on attention bias and attention control, as well as pain symptomatology (primary outcome), pain catastrophizing, anxiety and depression symptoms, and functional disability (secondary outcomes) immediately after training and 3 months later. We found no evidence that ABM changed attention bias or attention control in comparison with placebo training or no training. We also found that pain and pain-related outcomes were no different for those undergoing ABM compared with placebo training or no training when tested immediately after training or 3 months later. Overall, we found no evidence to support the efficacy of dot-probe ABM for improving pain-related outcomes in adolescents with chronic pain. This study was registered on the NIHR Clinical Research Network Portfolio in August 2014 (UK Clinical Trials Gateway: CPMS 17251) and funded by a Research Training Fellowship awarded to Lauren Heathcote by Action Medical Research for Children.


Genome-wide single nucleotide polymorphism analysis reveals recent genetic introgression from domestic pigs into Northwest European wild boar populations.
Present-day genetic introgression from domestic pigs into European wild boar has been suggested in various studies. However, no hybrids have been identified beyond doubt mainly because available methods were unable to quantify the extent of introgression and rule out natural processes. Genetic introgression from domestic pigs may have far-reaching ecological consequences by altering traits like the reproduction rate or immunology of wild boar. In this study, we demonstrate a novel approach to investigate genetic introgression in a Northwest (NW) European wild boar data set using a genome-wide single nucleotide polymorphism (SNP) assay developed for domestic pigs. We quantified the extent of introgression using allele frequency spectrum analysis, in silico hybridization simulations and genome distribution patterns of introgressed SNPs. Levels of recent introgression in the study area were expected to be low, as pig farming practices are prevailingly intensive and indoors. However, evidence was found for geographically widespread presence of domestic pig SNPs in 10% of analysed wild boar. This was supported by the identification of two different pig mitochondrial DNA haplotypes in three of the identified hybrid wild boar, suggesting that introgression had occurred from multiple sources (pig breeds). In silico hybridization simulations showed that the level of introgression in the identified hybrid wild boar is equivalent to first-generation hybrids until fifth-generation backcrosses with wild boar. The distribution pattern of introgressed SNPs supported these assignments in four of nine hybrids. The other five hybrids are considered advanced-generation hybrids, resulting from interbreeding among hybrid individuals. Three of nine hybrids were genetically associated with a different wild boar population than the one in which they were sampled. This discrepancy suggests that genetic introgression has occurred through the escape or release of an already hybridized farmed wild boar stock. We conclude that genetic introgression from domestic pigs into NW European wild boar populations is more recent and more common than expected and that genome-wide SNP analysis is a promising tool to quantify recent hybridization in free-living populations.


Functional dissection of Drosophila melanogaster SUUR protein influence on H3K27me3 profile.
In eukaryotes, heterochromatin replicates late in S phase of the cell cycle and contains specific covalent modifications of histones. SuUR mutation found in Drosophila makes heterochromatin replicate earlier than in wild type and reduces the level of repressive histone modifications. SUUR protein was shown to be associated with moving replication forks, apparently through the interaction with PCNA. The biological process underlying the effects of SUUR on replication and composition of heterochromatin remains unknown. Here we performed a functional dissection of SUUR protein effects on H3K27me3 level. Using hidden Markow model-based algorithm we revealed SuUR-sensitive chromosomal regions that demonstrated unusual characteristics: They do not contain Polycomb and require SUUR function to sustain H3K27me3 level. We tested the role of SUUR protein in the mechanisms that could affect H3K27me3 histone levels in these regions. We found that SUUR does not affect the initial H3K27me3 pattern formation in embryogenesis or Polycomb distribution in the chromosomes. We also ruled out the possible effect of SUUR on histone genes expression and its involvement in DSB repair. Obtained results support the idea that SUUR protein contributes to the heterochromatin maintenance during the chromosome replication. A model that explains major SUUR-associated phenotypes is proposed.


Incidence, Risk Factors, and Outcomes of Colorectal Cancer in Patients With Ulcerative Colitis With Low-Grade Dysplasia: A Systematic Review and Meta-analysis.
Little is known about outcomes of patients with ulcerative colitis with low-grade dysplasia (UC-LGD). We estimated the incidence of and risk factors for progression to colorectal cancer (CRC) in cohorts of patients with UC-LGD who underwent surveillance (surveillance cohort), and the prevalence of dysplasia-related findings among patients who underwent colectomy for UC-LGD (surgical cohort). We performed a systematic literature review through June 1, 2016, to identify cohort studies of adults with UC-LGD. We estimated pooled incidence rates of CRC and risk factors associated with dysplasia progression in surveillance cohorts, and prevalence of synchronous advanced neoplasia (CRC and/or high-grade dysplasia) in surgical cohorts. In 14 surveillance cohort studies of 671 patients with UC-LGD (52 developed CRC), the pooled annual incidence of CRC was 0.8% (95% confidence interval [CI], 0.4-1.3); the pooled annual incidence of advanced neoplasia was 1.8% (95% CI, 0.9-2.7). Risk of CRC was higher when LGD was diagnosed by expert gastrointestinal pathologist (1.5%) than by community pathologists (0.2%). Factors significantly associated with dysplasia progression were concomitant primary sclerosing cholangitis (odds ratio [OR], 3.4; 95% CI, 1.5-7.8), invisible dysplasia (vs visible dysplasia; OR, 1.9; 95% CI, 1.0-3.4), distal location (vs proximal location; OR, 2.0; 95% CI, 1.1-3.7), and multifocal dysplasia (vs unifocal dysplasia; OR, 3.5; 95% CI, 1.5-8.5). In 12 surgical cohort studies of 450 patients who underwent colectomy for UC-LGD, 34 patients had synchronous CRC (pooled prevalence, 17%; 95% CI, 8-33). In a systematic review of the literature, we found that among patients with UC-LGD under surveillance, the annual incidence of progression to CRC was 0.8%; differences in rates of LGD diagnosis varied with pathologists' level of expertise. Concomitant primary sclerosing cholangitis, invisible dysplasia, distal location, and multifocal LGD are high-risk features associated with dysplasia progression.


A functional network involved in the recycling of nucleocytoplasmic pre-60S factors.
Eukaryotic pre-ribosomes go through cytoplasmic maturation steps before entering translation. The nucleocytoplasmic proteins participating in these late stages of maturation are reimported to the nucleus. In this study, we describe a functional network focused on Rei1/Ybr267w, a strictly cytoplasmic pre-60S factor indirectly involved in nuclear 27S pre-ribosomal RNA processing. In the absence of Rei1, the nuclear import of at least three other pre-60S factors is impaired. The accumulation in the cytoplasm of a small complex formed by the association of Arx1 with a novel factor, Alb1/Yjl122w, inhibits the release of the putative antiassociation factor Tif6 from the premature large ribosomal subunits and its recycling to the nucleus. We propose a model in which Rei1 is a key factor for the coordinated dissociation and recycling of the last pre-60S factors before newly synthesized large ribosomal subunits enter translation.


Molecular analysis on the chemopreventive properties of resveratrol, a plant polyphenol microcomponent.
As a plant microcomponent, resveratrol is a polyphenolic compound produced by several species and found especially in Polygonum roots, peanuts seeds, berries and also grape and therefore can be present in human diet or beverages (red wine, for instance). Traditional chinese medicine and more recent epidemiological studies strongly suggested that resveratrol may act as a cancer chemopreventive compound. The biochemical mechanism by which resveratrol inhibits cell proliferation was provided by studies in numerous human cell lines including our work in hepatoblastoma HepG2 and colorectal tumor SW480 cells. The results show that resveratrol strongly inhibits cell proliferation at the micromolar range in a time- and dose-dependent manner. Resveratrol appears to block the cell cycle at the transition S to G2/M since there is no inhibition of [3H]-thymidine incorporation observed, while there is an increase of the cell number in S phase. On the other hand, in order to evaluate if the amount of resveratrol taken up during food or drink consumption is sufficient to ensure in the whole body the in vitro described beneficial effects, we evaluated the ratio between plasmatic level of resveratrol and its cell bioabsorption. Our study reports a higher uptake of resveratrol in the human hepatic derived HepG2 cells than in colorectal derived SW480 cells. In contrast, resveratrol is conjugated in these cells and derivatives are released in large amounts in the cell medium. Based on present knowledge, resveratrol appears to be a promising bioactive natural molecule with potential applications in phytotherapy, pharmacology or in nutriprotection (nutraceutic food) area.


Tissue metabolism driven arterial tree generation.
We present an approach to generate 3-D arterial tree models based on physiological principles while at the same time certain morphological properties are enforced at construction time. The driving force of the construction is a simplified angiogenesis model incorporating case-specific information about the metabolic demand within the considered domain. The vascular tree is constructed iteratively by successively adding new segments in chemotactic response to angiogenic growth factors secreted by ischemic cells. Morphometrically confirmed bifurcation statistics of vascular networks are incorporated to optimize the synthetic vasculature. The proposed method is able to generate artificial, yet physiologically plausible, arterial tree models that match the metabolic demand of the embedding tissue and fulfill the prescribed morphological properties at the same time. The proposed tree generation approach is applied in a simulation setup based on the metabolic configuration and anatomy of the macaque visual cortex. We analyze the generated tree models with respect to morphological and physiological aspects including fluid-dynamic simulations. The comparison of our results with the findings of different studies on the structure of cerebral vasculatures demonstrates the plausibility of our method.


Bone Mineral Density and Risk of Heart Failure in Older Adults: The Cardiovascular Health Study.
Despite increasing evidence of a common link between bone and heart health, the relationship between bone mineral density (BMD) and heart failure (HF) risk remains insufficiently studied. We investigated whether BMD measured by dual-energy x-ray absorptiometry was associated with incident HF in an older cohort. Cox models were stratified by sex and interactions of BMD with race assessed. BMD was examined at the total hip and femoral neck separately, both continuously and by World Health Organization categories. Of 1250 participants, 442 (55% women) developed HF during the median follow-up of 10.5 years. In both black and nonblack women, neither total hip nor femoral neck BMD was significantly associated with HF; there was no significant interaction by race. In black and nonblack men, total hip, but not femoral neck, BMD was significantly associated with HF, with evidence of an interaction by race. In nonblack men, lower total hip BMD was associated with higher HF risk (hazard ratio, 1.13 [95% CI, 1.01-1.26] per 0.1 g/cm2 decrement), whereas in black men, lower total hip BMD was associated with lower HF risk (hazard ratio, 0.74 [95% CI, 0.59-0.94]). There were no black men with total hip osteoporosis. Among nonblack men, total hip osteoporosis was associated with higher HF risk (hazard ratio, 2.83 [95% CI, 1.39-5.74]) compared with normal BMD. Among older adults, lower total hip BMD was associated with higher HF risk in nonblack men but lower risk in black men, with no evidence of an association in women. Further research is needed to replicate these findings and to study potential underlying pathways.


A cultural and social cognitive model of differences in acculturation orientations, alcohol expectancies, and alcohol-related risk behaviors among Hispanic college students.
The present study used a cultural and social cognitive conceptual framework to investigate whether alcohol expectancies and valuations would mediate the associations between specific acculturation orientations and alcohol-related risk behaviors. The sample comprised 1,527 Hispanic students attending colleges and universities in diverse regions of the United States. Respondents completed self-report measures of Hispanic and American cultural practices; alcohol expectancies and valuations; and self-reported frequency of hazardous alcohol use, binge drinking, sexual activity under the influence of alcohol, driving under the influence of alcohol, and riding with a drunk driver. Latent class analysis was used to classify participants into acculturation orientations. Results indicated that acculturation orientations were differentially associated with alcohol-related risk outcomes, with separated bicultural and low bicultural orientations inversely related to all of the alcohol-related risk behaviors except for riding with a drunk driver. Negative expectancy valuations were positively associated with endorsement of binge drinking and drunk driving and negative expectancies were negatively associated with binge drinking, drunk driving, and riding with a drunk driver. With the exception of sexual activity under the influence of alcohol, the associations between acculturation orientations and alcohol-related risk behaviors were partially mediated by positive alcohol expectancies. Our findings provided relevant data that are informative for preventing alcohol and related risk behaviors among Hispanic college students.


Three genes code for rubella virus structural proteins E1, E2a, E2b and C.
The structural proteins E1, E2a, E2b and C of rubella virus (RV) were purified by preparative SDS-PAGE. The individual proteins were subjected to amino-terminal sequence analysis by Edman degradation, carboxyl-terminal structure analysis by digestion with carboxypeptidases and quantitative amino acid composition analysis. The partial amino-terminal sequences of E2a and E2b were identical and different from that of E1. The C protein did not yield any consistent results on Edman degradation, suggesting that its amino-terminus is blocked. The amino acid compositions of E2a and E2b were very similar and differed from that obtained for E1 and C, which also differed from each other. Carboxypeptidase digestions showed that E2a and E2b have an identical carboxyl-terminal structure, which differed from that of the C protein. No amino acid residues were released from the E1 protein by digestion with a mixture of carboxypeptidases A and B. These results confirm that the structural proteins of RV are translated from three genes corresponding to C, E2 and E1. E2 exists in virions in two post-translationally modified forms, E2a and E2b, which have an identical apoprotein moiety. The partial amino acid sequence information obtained here should also be sufficient to localize the ends of the individual genes on the 24S mRNA genome once its nucleotide sequence has been established.


Anticipatory pacing strategies during supramaximal exercise lasting longer than 30 s.
This study assessed whether pacing strategies are adopted during supramaximal exercise bouts lasting longer than 30 s. Eight healthy males performed six Wingate anaerobic tests (WAnT). Subjects were informed that they were performing four 30-s WAnT, a 33-s, and a 36-s WAnT. However, they actually completed two trials of 30, 33, and 36 s each. Temporal feedback in the deception trials was manipulated so that subjects were unaware of the time discrepancy. Power output was determined from the angular displacement of the flywheel. The peak power (PPI), mean power (MPI), and fatigue (FI) indices were calculated for each trial. Power output was similar for all trials up to 30 s. However, at 36 s, the power output was significantly lower in the 36-s deception trial compared with the 36-s informed trial (392 +/- 32 W vs 470 +/- 88 W) (P < 0.001). The MPI was significantly lower in the 36-s trials (714 +/- 76 W and 713 +/- 78 W) compared with the 30-s trials (745 +/- 65 W and 764 +/- 82 W) although they were not different at 30 s (764 +/- 83 W and 755 +/- 79 W). The significant reduction in FI was greatest in the 36-s deception trial. The significant reduction in power output in the last 6 s of the 36-s deception trial, but not in the 36-s informed trial, indicates the presence of a preprogrammed 30-s "end point" based on the anticipated exercise duration from previous experience. The similarity in pacing strategy suggests that the pacing strategy is centrally regulated.


A kinetic analysis of protein transport through the anthrax toxin channel.
Anthrax toxin is composed of three proteins: a translocase heptameric channel, (PA(63))(7), formed from protective antigen (PA), which allows the other two proteins, lethal factor (LF) and edema factor (EF), to translocate across a host cell's endosomal membrane, disrupting cellular homeostasis. (PA(63))(7) incorporated into planar phospholipid bilayer membranes forms a channel capable of transporting LF and EF. Protein translocation through the channel can be driven by voltage on a timescale of seconds. A characteristic of the translocation of LF(N), the N-terminal 263 residues of LF, is its S-shaped kinetics. Because all of the translocation experiments reported in the literature have been performed with more than one LF(N) molecule bound to most of the channels, it is not clear whether the S-shaped kinetics are an intrinsic characteristic of translocation kinetics or are merely a consequence of the translocation in tandem of two or three LF(N)s. In this paper, we show both in macroscopic and single-channel experiments that even with only one LF(N) bound to the channel, the translocation kinetics are S shaped. As expected, the translocation rate is slower with more than one LF(N) bound. We also present a simple electrodiffusion model of translocation in which LF(N) is represented as a charged rod that moves subject to both Brownian motion and an applied electric field. The cumulative distribution of first-passage times of the rod past the end of the channel displays S-shaped kinetics with a voltage dependence in agreement with experimental data.


Neuroprotection with metformin and thymoquinone against ethanol-induced apoptotic neurodegeneration in prenatal rat cortical neurons.
Exposure to ethanol during early development triggers severe neuronal death by activating multiple stress pathways and causes neurological disorders, such as fetal alcohol effects or fetal alcohol syndrome. This study investigated the effect of ethanol on intracellular events that predispose developing neurons for apoptosis via calcium-mediated signaling. Although the underlying molecular mechanisms of ethanol neurotoxicity are not completely determined, mitochondrial dysfunction, altered calcium homeostasis and apoptosis-related proteins have been implicated in ethanol neurotoxicity. The present study was designed to evaluate the neuroprotective mechanisms of metformin (Met) and thymoquinone (TQ) during ethanol toxicity in rat prenatal cortical neurons at gestational day (GD) 17.5. We found that Met and TQ, separately and synergistically, increased cell viability after ethanol (100 mM) exposure for 12 hours and attenuated the elevation of cytosolic free calcium [Ca²⁺]c. Furthermore, Met and TQ maintained normal physiological mitochondrial transmembrane potential (ΔψM), which is typically lowered by ethanol exposure. Increased cytosolic free [Ca²⁺]c and lowered mitochondrial transmembrane potential after ethanol exposure significantly decreased the expression of a key anti-apoptotic protein (Bcl-2), increased expression of Bax, and stimulated the release of cytochrome-c from mitochondria. Met and TQ treatment inhibited the apoptotic cascade by increasing Bcl-2 expression. These compounds also repressed the activation of caspase-9 and caspase-3 and reduced the cleavage of PARP-1. Morphological conformation of cell death was assessed by TUNEL, Fluoro-Jade-B, and PI staining. These staining methods demonstrated more cell death after ethanol treatment, while Met, TQ or Met plus TQ prevented ethanol-induced apoptotic cell death. These findings suggested that Met and TQ are strong protective agents against ethanol-induced neuronal apoptosis in primary rat cortical neurons. The collective data demonstrated that Met and TQ have the potential to ameliorate ethanol neurotoxicity and revealed a possible protective target mechanism for the damaging effects of ethanol during early brain development.


Effect of n-3 fatty acids and antioxidants on oxidative stress after exercise.
n-3 fatty acids are known to exert multiple beneficial effects including anti-inflammatory actions that may diminish oxidative stress. Supplementation with antioxidant vitamins has been proposed to counteract oxidative stress and improve antioxidant status. Therefore, this project investigated the effects of daily supplementation in 48 trained cyclists over 6 wk and during 3 d of continuous exercise on F2-isoprostanes (oxidative stress), plasma n-3 fatty acids, and antioxidant status (oxygen radical absorption capacity and ferric-reducing antioxidant potential). Cyclists were randomized into n-3 fatty acids (N3) (n = 11) (2000 mg of eicosapentaenoic acid and 400 mg of docosahexaenoic acid), a vitamin-mineral (VM) complex (n = 12) emphasizing vitamins C (2000 mg), E (800 IU), A (3000 IU), and selenium (200 microg), a VM and n-3 fatty acid combination (VN3) (n = 13), or placebo (P) (n = 12). Blood was collected at baseline and preexercise and postexercise. A 4 x 3 repeated-measures ANOVA was performed to test main effects. After exercise, F2-isoprostanes were higher in N3 (treatment effect P = 0.014). Eicosapentaenoic acid and docosahexaenoic acid plasma values were higher after supplementation (interaction effect P = 0.001 and 0.006, respectively) in both n-3 supplemented groups. Oxygen radical absorption capacity declined similarly among all groups after exercise. Ferric-reducing antioxidant potential exhibited significant interaction (P = 0.045) and significantly increased after exercise in VN3 and VM (P < 0.01). This study indicates that supplementation with n-3 fatty acids alone significantly increases F2-isoprostanes after exhaustive exercise. Lastly, antioxidant supplementation augments plasma antioxidant status and modestly attenuates but does not prevent the significant n-3 fatty acid associated increase in F2-isoprostanes postexercise.


Origin and evolution of candidate mental retardation genes on the human X chromosome (MRX).
The human X chromosome has a biased gene content. One group of genes that is over-represented on the human X are those expressed in the brain, explaining the large number of sex-linked mental retardation (MRX) syndromes. To determine if MRX genes were recruited to the X, or whether their brain-specific functions were acquired after relocation to the mammalian X chromosome, we examined the location and expression of their orthologues in marsupials, which diverged from human approximately 180 million years ago. We isolated and mapped nine tammar wallaby MRX homologues, finding that six were located on the tammar wallaby X (which represents the ancient conserved mammal X) and three on chromosome 5, representing the recently added region of the human X chromosome. The location of MRX genes within the same synteny groups in human and wallaby does not support the hypothesis that genes with an important function in the brain were recruited in multiple independent events from autosomes to the mammalian X chromosome. Most of the tammar wallaby MRX homologues were more widely expressed in tammar wallaby than in human. Only one, the tammar wallaby ARX homologue (located on tammar chromosome 5p), has a restricted expression pattern comparable to its pattern in human. The retention of the brain-specific expression of ARX over 180 million years suggests that this gene plays a fundamental role in mammalian brain development and function. Our results suggest all the genes in this study may have originally had more general functions that became more specialised and important in brain function during evolution of humans and other placental mammals.


Simulation of the experimental imaging results for the OH + CHD3 reaction with a simple and accurate theoretical approach.
The OH + CHD3 reaction is among the largest one ever studied at the high-resolution level permitted by imaging techniques [B. Zhang et al., J. Phys. Chem. A, 2005, 109, 8989]. This process involves eighteen configuration space coordinates, which are large enough to make exact quantum scattering calculations beyond reach. Moreover, freezing some degrees-of-freedom in order to render these calculations feasible may lead to unrealistic predictions. However, we have found it possible to reproduce for the first time the pair-correlated measurements of Zhang et al. at a nearly quantitative level by means of full-dimensional classical trajectory calculations in a quantum spirit on a recent ab initio potential energy surface. These calculations combine the classical description of the dynamics, well suited to polyatomic systems, with Bohr quantization of both reagent and product vibrational motions. While this pseudo-quantization is exactly imposed to the reagents, it is approximately imposed to the products in a first step through energy-based Gaussian binning (1GB). In a second step, we show that the original action-based Gaussian binning (GB), long thought to be inapplicable in practice to polyatomic reactions, yields in fact results comparable in accuracy and numerical cost to those obtained by means of 1GB, provided that Gaussian weights are properly widened. This new finding clearly extends the scope of GB in theoretical reactive scattering.


Dopamine and training-related working-memory improvement.
Converging evidence indicates that the neurotransmitter dopamine (DA) is implicated in working-memory (WM) functioning and that WM is trainable. We review recent work suggesting that DA is critically involved in the ability to benefit from WM interventions. Functional MRI studies reveal increased striatal BOLD activity following certain forms of WM interventions, such as updating training. Increased striatal BOLD activity has also been linked to transfer of learning to non-trained WM tasks, suggesting a neural signature of transfer. The striatal BOLD signal is partly determined by DA activity. Consistent with this assertion, PET research demonstrates increased striatal DA release during updating of information in WM after training. Genetic studies indicate larger increases in WM performance post training for those who carry advantageous alleles of DA-relevant genes. These patterns of results corroborate the role of DA in WM improvement. Future research avenues include: (a) neuromodulatory correlates of transfer; (b) the potential of WM training to enhance DA release in older adults; (c) comparisons among different WM processes (i.e., updating, switching, inhibition) regarding regional patterns of training-related DA release; and (d) gene-gene interactions in relation to training-related WM gains.


Analytic derivation of the longitudinal component of the three-dimensional point-spread function in coded-aperture laminography.
Near-field coded-aperture data from a single view contain information useful for three-dimensional (3D) reconstruction. A common approach is to reconstruct the 3D image one plane at a time. An analytic expression is derived for the 3D point-spread function of coded-aperture laminography. Comparison with computer simulations and experiments for apertures with different size, pattern, and pattern family shows good agreement in all cases considered. The expression is discussed in the context of the completeness conditions for projection data and is applied to explain an example of nonlinear behavior inherent in 3D laminographic imaging.


A different form of long-lasting potentiation revealed in tissue plasminogen activator mutant mice.
The establishment of hippocampal long-term (LTP) requires protein and mRNA synthesis, suggesting that neuronal activity resulting in LTP initiates a cascade of gene expression. The expression of the gene for the extracellular serine protease tissue plasminogen activator (t-PA) is induced during LTP. Here we analyze long-lasting LTP (L-LTP,>4 hr)in CA1 hippocampal slices of mice homozygous for disrupted t-PA genes. Although mutant mice appear to exhibit long-term potentiation, we provide evidence that these mice are devoid of conventional homosynaptic L-LTP at the Schaffer collateral-CA1 pyramidal cell synapse. Most remarkably, t-PA-deficient mice exhibit a different form of long-lasting potentiation that is characterized by an NMDA receptor-dependent modification of GABA transmission in the CA1 region.


Reproductive trade-offs in a temperate reef fish under high pCO2 levels.
Fishes are currently facing novel types of anthropogenic stressors that have never experienced in their evolutionary history, such as ocean acidification. Under these stressful conditions, energetically costly processes, such as reproduction, may be sacrificed for increased chances of survival. This trade-off does not only affect the organism itself but may result in reduced offspring fitness. In the present study, the effects of exposure to high pCO2 levels were tested on the reproductive performance of a temperate species, the two-spotted goby, Gobiusculus flavescens. Breeding pairs were kept under control (∼600 μatm, pH∼ 8.05) and high pCO2 levels (∼2300 μatm, pH∼ 7.60) conditions for a 4-month period. Additionally, oxidative stress and energy metabolism-related biomarkers were measured. Results suggest that reproductive activity is stimulated under high pCO2 levels. Parental pairs in the simulated ocean acidification conditions exhibited increased reproductive output, with 50% more clutches and 44% more eggs per clutch than pairs under control conditions. However, there was an apparent trade-off between offspring number and size, as larvae of parental pairs under high pCO2 levels hatched significantly smaller, suggesting differences in parental provisioning, which could be related to the fact that these females produce more eggs. Moreover, results support the hypothesis of different energy allocation strategies used by females under high pCO2 conditions. These changes might, ultimately, affect individual fitness and population replenishment.


Mass cytometry deep phenotyping of human mononuclear phagocytes and myeloid-derived suppressor cells from human blood and bone marrow.
The monocyte phagocyte system (MPS) includes numerous monocyte, macrophage, and dendritic cell (DC) populations that are heterogeneous, both phenotypically and functionally. In this study, we sought to characterize those diverse MPS phenotypes with mass cytometry (CyTOF). To identify a deep phenotype of monocytes, macrophages, and DCs, a panel was designed to measure 38 identity, activation, and polarization markers, including CD14, CD16, HLA-DR, CD163, CD206, CD33, CD36, CD32, CD64, CD13, CD11b, CD11c, CD86, and CD274. MPS diversity was characterized for 1) circulating monocytes from healthy donors, 2) monocyte-derived macrophages further polarized in vitro (i.e., M-CSF, GM-CSF, IL-4, IL-10, IFN-γ, or LPS long-term stimulations), 3) monocyte-derived DCs, and 4) myeloid-derived suppressor cells (MDSCs), generated in vitro from bone marrow and/or peripheral blood. Known monocyte subsets were detected in peripheral blood to validate the panel and analysis pipeline. Then, using various culture conditions and stimuli before CyTOF analysis, we constructed a multidimensional framework for the MPS compartment, which was registered against historical M1 or M2 macrophages, monocyte subsets, and DCs. Notably, MDSCs generated in vitro from bone marrow expressed more S100A9 than when generated from peripheral blood. Finally, to test the approach in vivo, peripheral blood from patients with melanoma (n = 5) was characterized and observed to be enriched for MDSCs with a phenotype of CD14+HLA-DRlowS100A9high (3% of PBMCs in healthy donors, 15.5% in patients with melanoma, P < 0.02). In summary, mass cytometry comprehensively characterized phenotypes of human monocyte, MDSC, macrophage, and DC subpopulations in both in vitro models and patients.


DNA methylation of AHCY may increase the risk of ischemic stroke.
Genetic factors play an important role in the pathogenesis of ischemic stroke. Of these, epigenetic modifications provide a new direction for the study of ischemic stroke pathogenesis. This study aimed to determine the correlation between DNA methylation of the gene encoding S-adenosylhomocysteine hydrolase (AHCY) and the risk of ischemic stroke in 64 ischemic stroke patients and 138 patients with traumatic brain injury (control group). The methylation level of AHCY was analyzed using quantitative methylation-specific polymerase chain reaction. Statistically significant differences in AHCY methylation levels were observed between the case group [medians (interquartile range): 0.13% (0.09%, 0.27%)] and the control group [0.06% (0.00%, 0.17%), p < 0.0001], and these associations remained significant in both male (p = 0.003) and female (p = 0.0005) subjects. A subgroup analysis by age revealed a considerably higher percentage of methylated AHCY in the case group than the control group in all age groups (age < 60 years, p = 0.007; age ≥ 60 years, p < 0.0001). A receiver operating characteristic (ROC) curve analysis revealed a trend toward a role for AHCY methylation as an indicator of risk in all ischemic patients [area under the curve (AUC) = 0.70, p = 0.0001], male patients (AUC = 0.67, p = 0.004), and female patients (AUC = 0.75, p = 0.0002). Our study confirmed a significant association between the AHCY DNA methylation level and the risk of ischemic stroke, suggesting that this gene methylation pattern may be a potential diagnostic marker of ischemic stroke.


Behavioral and electrophysiological effects of cortical microstimulation parameters.
Electrical microstimulation has been widely used to artificially activate neural circuits on fast time scales. Despite the ubiquity of its use, little is known about precisely how it activates neural pathways. Current is typically delivered to neural tissue in a manner that provides a locally balanced injection of positive and negative charge, resulting in negligible net charge delivery to avoid the neurotoxic effects of charge accumulation. Modeling studies have suggested that the most common approach, using a temporally symmetric current pulse waveform as the base unit of stimulation, results in preferential activation of axons, causing diffuse activation of neurons relative to the stimulation site. Altering waveform shape and using an asymmetric current pulse waveform theoretically reverses this bias and preferentially activates cell bodies, providing increased specificity. In separate studies, measurements of downstream cortical activation from sub-cortical microstimulation are consistent with this hypothesis, as are recent measurements of behavioral detection threshold currents from cortical microstimulation. Here, we compared the behavioral and electrophysiological effects of symmetric vs. asymmetric current waveform shape in cortical microstimulation. Using a go/no-go behavioral task, we found that microstimulation waveform shape significantly shifts psychometric performance, where a larger current pulse was necessary when applying an asymmetric waveform to elicit the same behavioral response, across a large range of behaviorally relevant current amplitudes. Using voltage-sensitive dye imaging of cortex in anesthetized animals with simultaneous cortical microstimulation, we found that altering microstimulation waveform shape shifted the cortical activation in a manner that mirrored the behavioral results. Taken together, these results are consistent with the hypothesis that asymmetric stimulation preferentially activates cell bodies, albeit at a higher threshold, as compared to symmetric stimulation. These findings demonstrate the sensitivity of the pathway to varying electrical stimulation parameters and underscore the importance of designing electrical stimuli for optimal activation of neural circuits.


Prophylactic Sacral Dressing for Pressure Ulcer Prevention in High-Risk Patients.
Patients in intensive care units are likely to have limited mobility owing to hemodynamic instability and activity orders for bed rest. Bed rest is indicated because of the severity of the disease process, which often involves intubation, sedation, paralysis, surgical procedures, poor nutrition, low flow states, and poor circulation. These patients are predisposed to the development and/or the progression of pressure ulcers not only because of their underlying diseases, but also because of limited mobility and deconditioned states of health. To assess whether treating high-risk patients with a prophylactic sacral dressing decreases the incidence of unit-acquired sacral pressure ulcers. An evidence-based tool for identifying patients at high risk for pressure ulcers was used in 3 intensive care units at an urban tertiary care hospital and academic medical center. Those patients deemed at high risk had a prophylactic sacral dressing applied. Incidence rates were collected and compared for the 7 months preceding use of the dressings and for 7 months during the trial period when the dressing was used. After the sacral dressing began being used, the number of unit-acquired sacral pressure ulcers decreased by 3.4 to 7.6 per 1000 patient days depending on the unit. A prophylactic sacral dressing may help prevent unit-acquired sacral pressure ulcers. Implementation of an involved care team with heightened awareness and increased education along with a prophylactic sacral dressing in patients deemed high risk for skin breakdown are all essential for success.


Exploring the electrochemical performance of graphite and graphene paste electrodes composed of varying lateral flake sizes.
We report the fabrication, characterisation (SEM/EDX, TEM, XRD, XPS and Raman spectroscopy) and electrochemical properties of graphite and graphene paste electrodes with varying lateral flake sizes. The fabricated paste electrodes are electrochemically analysed using both outer-sphere and inner-sphere redox probes, namely; hexaammineruthenium(iii) chloride, N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), potassium ferrocyanide(ii) and ammonium ferrous(ii) sulphate. Upon comparison of different graphite paste electrodes, a clear correlation between the lateral flake sizes (La), ranging from 1.5 mm-0.5 μm, and electrochemical activity (heterogeneous electron transfer (HET) kinetics) is evident, where an improvement in the HET is observed at smaller lateral flake sizes. We infer that the beneficial response evident when employing laterally smaller flakes is due to an increased number of edge plane like-sites/defects available upon the electrode surface, facilitating electron transfer. Interestingly, given that the overall lateral flake sizes of the graphenes utilised (10.0-1.3 μm) were significantly smaller than those studied previously, an improvement in HET kinetics was also evident with the reduction of lateral flake size; the extent to which is redox-probe dependent. Improvements are observed up to a distinct point, termed the 'lateral size threshold' (ca. ≤2 μm) where the electrochemical reversible limit is approached. Further support is provided from density functional theory (DFT), exploring the electronic structure (i.e. HOMO-LUMO) as a function of flake size, which demonstrates that the coverage of edge plane like-sites/defects comprising the geometric structure of the relatively small graphene flakes is such that effectively the entire flake has become electrochemically active. In this study, the importance of lateral flake size with respect to electrochemical reactivity at carbon-based electrodes has been demonstrated alongside a structural relationship upon HET performance, a phenomenon that has not previously been described in the literature. Such work is both highly important and informative for the field of electrochemistry and electrode performance, with potential implications in a plethora of areas, ranging from novel renewable energy sources to electroanalytical sensing platforms.


A new PCR-based bioassay strategy for the detection of type A trichothecenes in food.
Type A trichothecenes (primarily T-2 and HT-2 toxins) are common fungal metabolites found in a wide range of grains and other field crops grown in temperate climatic zones. By acting as potent inhibitors of protein synthesis, T-2 and HT-2 exert adverse effects particularly against rapidly proliferating tissues, including the bone marrow, the immune system and epithelial cells. Based on toxicity studies in laboratory and farm animals, a temporary tolerable daily intake for the sum of T-2 and HT-2 has been issued in the European Union. However, exposure assessments suggest that the combined intake of these natural compounds exceeds in many cases the proposed threshold. To further protect the consumers, it is therefore necessary to screen a large number of food samples for parts per billion levels of both T-2 and HT-2. Towards that goal, we are the first to report that these two type A trichothecenes induce fast and high-amplitude transcriptional changes in cultured human breast cancer cells. This specific response involving marker gene inductions by more than 1000-fold has been exploited to develop a real-time PCR-based screening method that displays a limit of detection of 5 ng g(-1) for T-2 and 10 ng g(-1) for HT-2. The practicability of this bioassay is demonstrated by its application to the detection of type A trichothecenes in different food matrices.


The cellular cancer resistance of the SR/CR mouse.
The SR/CR mouse phenotype, first described in 1999 in BALB/c and later bred into C57BL/6 mice, is resistant to cancer formation following high doses of cancer cells administered intraperitoneally. The tumor cell targeting and destruction mechanisms have not been identified. By fluorescence-activated cell sorting analysis, the immune response of SR/CR mice after intraperitoneal injection of cancer cells was investigated and compared with parent strain mice. A massive influx of leukocytes into the peritoneal cavity was found. A large fraction of these leukocytes were polymorphonuclear granulocytes, macrophages and natural killer cells. A relative decrease in influx of B-cells compared with controls was demonstrated. Increased proportions of leukocytes belonging to the innate immune system were also demonstrated in splenocytes of SR/CR mice. Cytospins of peritoneal fluid from SR/CR mice after cancer cell injection showed formations of immune cells morphologically resembling polymorphonuclear granulocytes and macrophages adjoining the cancer cells. The results point to the potential involvement of innate immune cells in cancer immunology. Our data support migration of polymorphonuclear granulocytes, macrophages and NK cells into the peritoneum of the SR/CR mouse in response to intraperitoneal injection of S180 cancer cells. The cell composition of spleens of SR/CR mice reflected the differential regulation of the innate immune cells in peritoneal exudates. Both peritoneal exudates and the spleens of SR/CR mice contained decreased proportions of B-cells compared with BALB/c and C57BL/6 mice. We reproduce important aspects of previous published data and further extend them by showing differentially regulated populations of splenocytes including B-lymphocytes in SR/CR mice compared with parent strain controls. Importantly, this differentially regulated immune response of SR/CR mice could not be found in response to challenge with the lymphoma cell line EL-4.


Brassinosteroid Involvement in Arabidopsis thaliana Stomatal Opening.
Stomata within the plant epidermis regulate CO2 uptake for photosynthesis and water loss through transpiration. Stomatal opening in Arabidopsis thaliana is determined by various factors, including blue light as a signal and multiple phytohormones. Plasma membrane transporters, including H+-ATPase, K+ channels and anion channels in guard cells, mediate these processes, and the activities and expression levels of these components determine stomatal aperture. However, the regulatory mechanisms involved in these processes are not fully understood. In this study, we used infrared thermography to isolate a mutant defective in stomatal opening in response to light. The causative mutation was identified as an allele of the brassinosteroid (BR) biosynthetic mutant dwarf5. Guard cells from this mutant exhibited normal H+-ATPase activity in response to blue light, but showed reduced K+ accumulation and inward-rectifying K+ (K+in) channel activity as a consequence of decreased expression of major K+in channel genes. Consistent with these results, another BR biosynthetic mutant, det2-1, and a BR receptor mutant, bri1-6, exhibited reduced blue light-dependent stomatal opening. Furthermore, application of BR to the hydroponic culture medium completely restored stomatal opening in dwarf5 and det2-1 but not in bri1-6. However, application of BR to the epidermis of dwarf5 did not restore stomatal response. From these results, we conclude that endogenous BR acts in a long-term manner and is required in guard cells with the ability to open stomata in response to light, probably through regulation of K+in channel activity.


The usage, occurrence and dietary intakes of white mineral oils and waxes in Europe.
Dietary exposures to mineral hydrocarbons were estimated from information about patterns of usage, concentrations in foods and quantities of foods consumed. An industry survey showed that the largest usage of food-grade white mineral oils was in plastics manufacture although the majority are used in non-food applications. The largest volumes of wax usage were in packaging. Conservative estimates indicated that daily intakes of white mineral oils ranged from 0.39 to 0.91 mg/kg bw/day for adults and from 0.75 to 1.77 mg/kg bw/day for children (mean and 97.5th percentiles). Total wax intakes ranged from 0.08 to 0.19 mg/kg bw/day for adults and 0.23 to 0.64 mg/kg bw/day for pre-school children. When usage factors were applied, estimates of chronic intakes of white oils were reduced to 0.09-0.20 mg/kg bw/day for adults and to 0.17-0.39 mg/kg bw/day for children. Total wax intakes were reduced to 0.01-0.02 mg/kg bw/day for adults and to 0.02-0.06 mg/kg bw/day for children. For white mineral oils the principal source of exposure for most consumers was imported de-dusted grain. The principal source of potential wax exposure was from glazing agents on confectionery. There was no evidence of intakes exceeding SCF ADIs for microcrystalline waxes or certain white mineral oils and levels of exposure were similar to those of naturally-occurring mineral hydrocarbons in foods.


Redefining Death: Urgent Need to Evolve toward a Homogeneous Definition of Death in India.
The interdependent circulatory-respiratory criteria and brainstem criteria are used to define death. Continuing life-support in brainstem dead patients is a legal conundrum for the health-care professionals and an ethical worry for the society at large. There is an urgent need to revisit this subject to evolve a more uniform definition of death irrespective of the circumstances. A compelling take-home message from this discussion is to spread awareness about brainstem death amongst healthcare professionals. Vadi S, Shroff S. Redefining Death: Urgent Need to Evolve Toward a Homogeneous Definition of Death in India. Indian J Crit Care Med 2019;23(8):368-370.


Medical home transformation: a gradual process and a continuum of attainment.
The patient-centered medical home is often discussed as though there exist either traditional practices or medical homes, with marked differences between them. We analyzed data from an evaluation of certified medical homes in Minnesota to study this topic. We obtained publicly reported composite measures for quality of care outcomes pertaining to diabetes and vascular disease for all clinics in Minnesota from 2008 to 2010. The extent of and change in practice systems over that same time period for the first 120 clinics serving adults certified as health care homes (HCHs) was measured by the Physician Practice Connections Research Survey (PPC-RS), a self-report tool similar to the National Committee for Quality Assurance standards for patient-centered medical homes. Measures were compared between these clinics and 518 non-HCH clinics in the state. Among the 102 clinics for which we had precertification and postcertification scores for both the PPC-RS and either diabetes or vascular disease measures, the mean increase in systems score over 3 years was an absolute 29.1% (SD = 16.7%) from a baseline score of 38.8% (SD = 16.5%, P ≤.001). The proportion of clinics in which all patients had optimal diabetes measures improved by an absolute 2.1% (SD = 5.5%, P ≤.001) and the proportion in which all had optimal cardiovascular disease measures by 4.4% (SD = 7.5%, P ≤.001), but all measures varied widely among clinics. Mean performance rates of HCH clinics were higher than those of non-HCH clinics, but there was extensive overlap, and neither group changed much over this time period. The extensive variation among HCH clinics, their overlap with non-HCH clinics, and the small change in performance over time suggest that medical homes are not similar, that change in outcomes is slow, and that there is a continuum of transformation.


Nurses' perception of the strategies to gaining professional power: A qualitative study.
Power in nursing is a broad concept that has a determining effect on the achievement of professional goals. Gaining power is essential for promoting the roles of nurses, improving their professional image and the consistent improvement of healthcare systems. This study was conducted to identify and clarify strategies for gaining power in the nursing profession through the experiences of Iranian nurses. The present qualitative grounded theory study was conducted on fifteen participants selected through purposive sampling. Data were collected through individual, in-depth, semi-structured interviews. The data obtained were analyzed using the guidelines provided by Corbin and Strauss, (2008 edition). This study was carried out in Qazvin city of Iran and lasted fourteen months (2015-2016). The analysis of the data were classified under the main theme of gaining human-professional power based on individual and organizational capacities, divided into four main categories, including respecting human values and ethical principles (with two subcategories, keeping the human symbols of power and commitment to moral obligations of power), promoting professional interactions (with two subcategories, paying attention to intraprofessional communication and paying attention to interprofessional communication), attempting professional endurance (with two subcategories, raising self-confidence and having professional commitment), and valuing potential capacities (with two subcategories, regard for individual capacities and regard for organizational capacities). The findings obtained suggest that success in gaining power in nursing, requires a cumulative focus on human, ethical, professional, individual and organizational capacities.


Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth.
Reactive oxygen species (ROS) including nitric oxide (NO) and superoxide anion (O(2)(-)) are associated with cell migration, proliferation and many growth-related diseases. The objective of this study was to determine whether there was a reciprocal relationship between rat coronary microvascular endothelial cell (CMEC) growth and activity/expressions (mRNA and protein) of endothelial NO synthase (eNOS) and NAD(P)H oxidase enzymes. Proliferating namely, 50% confluent CMEC possessed approximately threefold increased activity and expression of both enzymes compared to 100% confluent cells. Treatment of CMEC with an inhibitor of eNOS (L-NAME, 100 microM) increased cell proliferation as assessed via three independent methods, i.e. cell counting, determination of total cellular protein levels and [(3)H]-thymidine incorporation. Similarly, treatment of CMEC with pyrogallol (0.3-3 mM), a superoxide anion (O(2)(-)) generator, also increased CMEC growth while spermine NONOate (SpNO), a NO donor, significantly reduced cell growth. Co-incubation of CMEC with a cell permeable superoxide dismutase mimetic (Mn-III-tetrakis-4-benzoic acid-porphyrin; MnTBAP) plus either pyrogallol or NO did not alter cell number and DNA synthesis thereby dismissing the involvement of peroxynitrite (OONO(-)) in CMEC proliferation. Specific inhibitors of NAD(P)H oxidase but not other ROS-generating enzymes including cyclooxygenase and xanthine oxidase, attenuated cell growth. Transfection of CMEC with antisense p22-phox cDNA, a membrane-bound component of NAD(P)H oxidase, resulted in substantial reduction in [(3)H]-thymidine incorporation, total cellular protein levels and expression of p22-phox protein. These data demonstrate a cross-talk between CMEC growth and eNOS and NAD(P)H oxidase enzyme activity and expression, thus suggesting that the regulation of these enzymes may be critical in preventing the initiation and/or progression of coronary atherosclerosis.


Application of a bromodeoxyuridine-Hoechst/ethidium bromide technique for the analysis of radiation-induced cell cycle delays in asynchronous cell populations.
A flow cytometric technique utilizing the continuous incorporation of bromodeoxyuridine (BrdU) into asynchronous cells to measure radiation-induced cell cycle delay is described. Following the incorporation of the BrdU label the cells are stained with ethidium bromide and the bis-benzimidazole Hoechst 33258. These fluorochromes have differential staining patterns. Hoechst 33258 fluoresces blue and is quenched by BrdU incorporated into cellular DNA during S phase. Ethidium bromide fluoresces red and is not quenched by BrdU. Therefore in cells that are cycling and synthesizing DNA new G1 and G2 compartments are created and this can be used to measure cell cycle delays following ionizing radiation to asynchronous cells. We have used this technique to evaluate two cell lines: a normal diploid human embryo fibroblast cell line MRC 5, which has inducible p53 and shows delays at both G1 and G2 checkpoints, and the human cervix carcinoma cell line HX 156. This cell line has been infected with human papilloma virus (HPV) 16, and therefore has inactivated p53 function and is blocked only at the G2 checkpoint. Using this method, cell cycle-dependent effects relating to the G2 block can be observed. The radiation-induced G2 block differs from that induced by drugs or heating in that cells are blocked in G2 irrespective of the phase of the cell cycle they are treated in. This method allows these different types of G2 block to be quantified.


First line targeted therapies in breast cancer: focus on bevacizumab.
The heterogeneity of metastatic breast cancer mandates the need to select therapies taking into account tumor and patient characteristics. Chemotherapy is indicated in the palliative setting especially when the disease is unresponsive to hormonal therapy or is hormone-receptor negative. The main chemotherapeutic agents are anthracyclines, taxanes, and capecitabine. The knowledge of the effects of currently approved agents and of the biology of breast cancer have paved the way for the evaluation of new treatment options, among which are anti-angiogenic agents. Angiogenesis inhibition has resulted in clinically significant improvements in the outcome of a variety of malignancies, including breast cancer. Bevacizumab, a monoclonal antibody anti-vascular endothelial growth factor (VEGF), is the most extensively studied anti-angiogenic compound. According to the results of a phase III trial in patients with untreated metastatic breast cancer, bevacizumab increases both objective response rate and median progression-free survival when combined with standard chemotherapy vs chemotherapy alone. The combination of anti-angiogenic drugs and other biologic agents is also being explored in an attempt to improve efficacy.


CD4+ T cell expression of MyD88 is essential for normal resolution of Chlamydia muridarum genital tract infection.
Resolution of Chlamydia genital tract infection is delayed in the absence of MyD88. In these studies, we first used bone marrow chimeras to demonstrate a requirement for MyD88 expression by hematopoietic cells in the presence of a wild-type epithelium. Using mixed bone marrow chimeras we then determined that MyD88 expression was specifically required in the adaptive immune compartment. Furthermore, adoptive transfer experiments revealed that CD4(+) T cell expression of MyD88 was necessary for normal resolution of genital tract infection. This requirement was associated with a reduced ability of MyD88(-/-)CD4(+) T cells to accumulate in the draining lymph nodes and genital tract when exposed to the same inflammatory milieu as wild-type CD4(+) T cells. We also demonstrated that the impaired infection control we observed in the absence of MyD88 could not be recapitulated by deficiencies in TLR or IL-1R signaling. In vitro, we detected an increased frequency of apoptotic MyD88(-/-)CD4(+) T cells upon activation in the absence of exogenous ligands for receptors upstream of MyD88. These data reveal an intrinsic requirement for MyD88 in CD4(+) T cells during Chlamydia infection and indicate that the importance of MyD88 extends beyond innate immune responses by directly influencing adaptive immunity.


Gonadotrophin therapy for ovulation induction in subfertility associated with polycystic ovary syndrome.
Approximately 15% of patients with PCOS remain anovulatory despite treatment with oral anti-oestrogen medications such as clomiphene citrate. In addition, about half of women with PCOS ovulating on anti-oestrogen treatment fail to conceive. Gonadotrophin stimulation is the next step in treatment for women who are "clomiphene resistant", however, results of gonadotrophin stimulation in women with PCOS are less successful. In PCOS associated with hypersecretion of LH, purified urinary follicle-stimulating hormone (u-FSH) preparations have theoretical advantages over the use of human menopausal gonadotrophin (hMG) preparations (containing both FSH and LH), but whether this claimed advantage extends into clinical practice remains uncertain. In addition, the use of gonadotrophin-releasing hormone analogues (GnRH-a) to produce pituitary desensitisation prior to ovulation induction in PCOS has been claimed to increase the success rates of treatment as well as reduce complications such as OHSS and multiple pregnancy. Gonadotrophin preparations have also been administered via different routes (intramuscular or subcutaneous), or using different stimulation regimens and protocols (step-up or standard) in an attempt to improve efficacy. To determine the effectiveness of urinary-derived gonadotrophins as ovulation induction agents in patients with PCOS trying to conceive. In particular, to assess the effectiveness of (1) different gonadotrophin preparations, (2) the addition of a gonadotrophin-releasing hormone agonist (GnRH-a) to gonadotrophin stimulation and (3) different modalities of gonadotrophin administration. The search strategy to identify RCTs consisted of (1) the Group's Specialised Register of Controlled Trials using the search strategy developed for the Menstrual Disorders and Subfertility Group as a whole (see the Review Group details for more information), (2) additional specific electronic Medline searches and (3) bibliographies of identified studies and narrative reviews. RCTs in which urinary-derived gonadotrophins were used for ovulation induction in patients with primary or secondary subfertility attributable to PCOS. Twenty three RCTs were identified, 9 of which were excluded from analysis. The data were extracted independently by 2 authors. The following criteria were assessed: (1) the methodological characteristics of the trials, (2) the baseline characteristics of the studied groups and (3) the outcomes of interest: pregnancy rate (per cycle), ovulation rate (per cycle), miscarriage rate (per pregnancy), multiple pregnancy rate (per pregnancy), overstimulation rate (per cycle) and ovarian hyperstimulation syndrome (OHSS) rate (per cycle). Where suitable, meta-analysis was performed using Peto's OR with 95% CI with the fixed effect Mantel-Haentszel equation. (1) A reduction in the incidence of OHSS with FSH compared to hMG in stimulation cycles without the concomitant use of a GnRH-a (OR 0.20; 95% CI 0.08-0.46) and (2) a higher overstimulation rate when a GnRH-a is added to gonadotrophins (OR 3.15; 95% CI 1.48-6.70). Although 14 RCTs were included in this review, few dealt with the same comparisons, all were small to moderate size and their methodological quality was generally poor. Any conclusions, therefore, remain tentative as they are based on a limited amount of data and will require further RCTs to substantiate them. In none of the comparisons was there a significant improvement in pregnancy rate but this may be due to the lack of power (i.e. insufficient patients randomised to demonstrate a significant difference between treatments). There was a trend towards better pregnancy rates with the addition of a GnRH-a to gonadotrophin stimulation and these interventions warrant further study. Despite theoretical advantages, urinary-derived FSH preparations did not improve pregnancy rates when compared to traditional and cheaper hMG preparations; their only demonstrable benefit was a reduced risk of OHSS in cycles when administered without the concomitant use of a GnRH-a. No conclusions can be drawn on miscarriage and multiple pregnancy rates due to insufficient reporting of these outcomes in the trials.


Gestational age at delivery of twins and perinatal outcomes: a cohort study in Aberdeen, Scotland.
Background: Twin pregnancy is associated with a threefold increase in perinatal death compared to singletons. The objective of this study was to determine the risk of perinatal death in twins by week of gestation and to quantify the effect of known risk factors. Methods: A cohort analysis was performed using data from the Aberdeen Maternity and Neonatal Databank (AMND). The exposure was gestational age at delivery and the primary outcome was perinatal death. Adjusted hazard ratios (aHRs) for perinatal death according to gestational age at delivery were determined by multivariate Cox proportional hazards regression modelling with robust standard errors to account for clustering in the twin infants. Confounders and risk factors quantified and adjusted for in the model included maternal age, smoking, parity, marital status and year of birth. Kaplan-Meier time to event analysis was used to determine the differences in survival according to chorionicity and assisted reproduction technologies (ART) conception status. Results: The population comprised of 7,420 twin babies born between 1950 and 2013 in the Grampian area of Northern Scotland. There were 272 stillbirths in the cohort (3.67%) and 273 neonatal deaths (3.68%). Compared to delivery at 37-38 weeks, delivery before 37 weeks was associated with a 2-fold increase in perinatal death. Monochorionic twins had a 2-fold increase in perinatal death compared to dichorionic twins (aHR 2.15, 95% CI 1.60-2.90). Twins conceived by ART did not have a greater risk of perinatal death compared to those naturally conceived (aHR 1.21, 95% CI 0.87-1.68) Conclusion: This study suggests that delivery of twins at 37-38 weeks is associated with the lowest risk of perinatal death.


Use of transrectal ultrasonography for assessment of the size and location of prostatic carcinoma in dogs.
To evaluate the use of transrectal ultrasonography (TRUS) for the assessment of prostatic tumors in dogs and to compare results for TRUS with results for other imaging modalities. 10 client-owned male dogs. Client-owned dogs identified with prostatic carcinoma were enrolled. Fluoroscopy, transabdominal ultrasonography (TAUS), TRUS, and MRI were performed on all dogs. Tumor measurements, urethral penetration (identification of abnormal tissue within the urethral lumen), and tumor extension into the urinary tract were recorded for all imaging modalities. Agreement between results for MRI (considered the criterion-referenced standard) and results for other modalities were compared. Median body weight of the 10 dogs was 26.3 kg (range, 9.4 to 49.5 kg). No complications were encountered during or after TRUS. Significant moderate to good agreements (intraclass correlation coefficients, 0.60 to 0.86) among TAUS, TRUS, fluoroscopy, and MRI were identified for tumor length and height. Assessments of urethral penetration and tumor extension into the bladder with TRUS did not differ significantly from those made with MRI and were superior in terms of absolute agreement with MRI when compared with those for TAUS. TRUS was successfully and safely used to evaluate prostatic carcinoma in dogs. There was moderate to good agreement with MRI results for tumor height and length measurements, and TRUS was found to be superior to TAUS for some assessments. Transrectal ultrasonography can be considered an adjunctive imaging modality for the performance of prostatic interventional procedures or assessment of response to treatment.


Resection of isolated local and metastatic recurrence in periampullary adenocarcinoma.
The majority of patients with periampullary cancer develop local or metastatic recurrence despite successful negative margin resection. Unfortunately, there are no established therapeutic strategies for managing these patients. The literature on the surgical resection of recurrent disease is limited. This is a retrospective study evaluating patients who underwent reoperative resection of recurrent periampullary cancer at a single institution between 1990 and 2011. Perioperative outcomes were compared with those of the original primary resections for patients with local recurrence. Kaplan-Meier curves were used to evaluate survival. Twenty-two patients underwent reoperative resection following the successful primary resection of periampullary cancers. Median survival from the time of reoperation was 28.1 months. A greater survival benefit was seen in patients undergoing reoperative resection with >15 months between the primary resection and recurrence (40.6 months versus 8.2 months; P < 0.05). Complication rates were lower after reoperative resection compared with the primary resection (20% versus 70%). Perioperative characteristics including operative time, estimated blood loss and hospital stay were similar in both the primary and reoperation procedures. Surgical resection of periampullary cancer recurrence is feasible, safe and may offer survival benefits in comparison with alternative treatment modalities. Reoperative resection should be considered, especially in patients in whom the time to recurrence is lengthy.


Feasibility and safety of augmented-reality glass for computed tomography-assisted percutaneous revascularization of coronary chronic total occlusion: A single center prospective pilot study.
Percutaneous coronary intervention (PCI) of chronic total occlusion (CTO) may be facilitated by projection of coronary computed tomography angiography (CTA) datasets in the catheterization laboratory. There is no data on the feasibility and safety outcomes of CTA-assisted CTO PCI using a wearable augmented-reality glass. A total of 15 patients scheduled for elective antegrade CTO intervention were prospectively enrolled and underwent preprocedural coronary CTA. Three-dimensional and curved multiplanar CT reconstructions were transmitted to a head-mounted hands-free computer worn by interventional cardiologists during CTO PCI to provide additional information on CTO tortuosity and calcification. The results of CTO PCI using a wearable computer were compared with a time-matched prospective angiographic registry of 59 patients undergoing antegrade CTO PCI without a wearable computer. Operators' satisfaction was assessed by a 5-point Likert scale. Mean age was 64 ± 8 years and the mean J-CTO score was 2.1 ± 0.9 in the CTA-assisted group. The voice-activated co-registration and review of CTA images in a wearable computer during CTO PCI were feasible and highly rated by PCI operators (4.7/5 points). There were no major adverse cardiovascular events. Compared with standard CTO PCI, CTA-assisted recanalization of CTO using a wearable computer showed more frequent selection of the first-choice stiff wire (0% vs 40%, p < 0.001) and lower contrast exposure (166 ± 52 vs 134 ± 43 ml, p = 0.03). Overall CTO success rates and safety outcomes remained similar between both groups. CTA-assisted CTO PCI using an augmented-reality glass is feasible and safe, and might reduce the resources required for the interventional treatment of CTO.


Recent approaches to the prioritization of candidate disease genes.
Many efforts are still devoted to the discovery of genes involved with specific phenotypes, in particular, diseases. High-throughput techniques are thus applied frequently to detect dozens or even hundreds of candidate genes. However, the experimental validation of many candidates is often an expensive and time-consuming task. Therefore, a great variety of computational approaches has been developed to support the identification of the most promising candidates for follow-up studies. The biomedical knowledge already available about the disease of interest and related genes is commonly exploited to find new gene-disease associations and to prioritize candidates. In this review, we highlight recent methodological advances in this research field of candidate gene prioritization. We focus on approaches that use network information and integrate heterogeneous data sources. Furthermore, we discuss current benchmarking procedures for evaluating and comparing different prioritization methods.


Informed Consent and Cognitive Dysfunction After Noncardiac Surgery in the Elderly.
Cognitive dysfunction 3 months after noncardiac surgery in the elderly satisfies informed consent thresholds of foreseeability in 10%-15% of patients, and materiality with new deficits observed in memory and executive function in patients with normal test performance beforehand. At present, the only safety step to avoid cognitive dysfunction after surgery is to forego surgery, thereby precluding the benefits of surgery with removal of pain and inflammation, and resumption of normal nutrition, physical activity, and sleep. To assure that consent for surgery is properly informed, risks of both cognitive dysfunction and alternative management strategies must be discussed with patients by the surgery team before a procedure is scheduled.


Gate-controlled electron transport in coronenes as a bottom-up approach towards graphene transistors.
Graphene is considered to be a large aromatic molecule, the limiting case of the family of polycyclic aromatic hydrocarbons. This fascinating two-dimensional material has many potential applications, including field effect transistors (FETs). However, the graphene sheets in these devices have irregular shapes and variable sizes, and contain various impurities and defects, which are undesirable for applications. Moreover, the bandgap of graphene is zero and, consequently, the on/off ratios of graphene FETs are small, making it difficult to build logic circuits. To overcome these difficulties, we report here a bottom-up attempt to fabricate nanoscale graphene FETs. We synthesize structurally well-defined coronene molecules (consisting of 13 benzene rings) terminated with linker groups, bridge each molecule to source and drain electrodes through the linkers, measure conductance and demonstrate the FET behaviour of the molecule.


On optimally combining pieces of information, with application to estimating 3-d complex-object position from range data.
New asymptotic methods are introduced that permit computationally simple Bayesian recognition and parameter estimation for many large data sets described by a combination of algebraic, geometric, and probabilistic models. The techniques introduced permit controlled decomposition of a large problem into small problems for separate parallel processing where maximum likelihood estimation or Bayesian estimation or recognition can be realized locally. These results can be combined to arrive at globally optimum estimation or recognition. The approach is applied to the maximum likelihood estimation of 3-D complex-object position. To this end, the surface of an object is modeled as a collection of patches of primitive quadrics, i.e., planar, cylindrical, and spherical patches, possibly augmented by boundary segments. The primitive surface-patch models are specified by geometric parameters, reflecting location, orientation, and dimension information. The object-position estimation is based on sets of range data points, each set associated with an object primitive. Probability density functions are introduced that model the generation of range measurement points. This entails the formulation of a noise mechanism in three-space accounting for inaccuracies in the 3-D measurements and possibly for inaccuracies in the 3-D modeling. We develop the necessary techniques for optimal local parameter estimation and primitive boundary or surface type recognition for each small patch of data, and then optimal combining of these inaccurate locally derived parameter estimates in order to arrive at roughly globally optimum object-position estimation.


Spatial heterogeneities of current and future hurricane flood risk along the U.S. Atlantic and Gulf coasts.
We evaluate the spatial heterogeneities of hurricane flood risk along the United States (U.S.) Atlantic and Gulf coasts under two different climate scenarios (current and future). The flood hazard is presented as the hurricane surge flood level with 1% annual exceedance probability (100-year flood) under the two scenarios, where the future scenario considers the effect of hurricane climatology change and sea level rise towards late-21st-century under a high emission scenario (RCP8.5). This hazard information is combined with estimated vulnerability and disaster resilience of coastal communities to map the relative current and future risk employing different risk definitions. Several geographical techniques and spatial distributional models (e.g., spatial autocorrelation, spatial hotspot analysis, and spatial multivariate clustering analysis) are applied to systematically analyze the risk and identify statistically significant hotspots of the highest risk. Most of the high-risk hotspots are found in the Gulf coast region, particularly along the west coast of Florida. However, two out of three risk evaluation approaches also indicate New York City as a risk hotspot under the future climate-showing that the resultant risk is sensitive to the consideration of evaluation factors (i.e., hazard, vulnerability, and resilience). Additionally, we apply a machine-learning algorithm-based spatial multivariate approach to map the spatially distinct groups based on the values of risk, hazard, vulnerability, and resilience. The results show that the counties in the highest risk group (value >3rd quartile, 15% of total counties, including New York City) in the future lack specifically in the community capital and the social components of community resilience. This assessment of coastal risk to hurricane flood has important policy-relevant implications to provide a focus-for-action for risk reduction and resilience enhancement for the U.S., where 6.5 million households live in the hurricane flood-prone areas.


Biofabrication: an overview of the approaches used for printing of living cells.
The development of cell printing is vital for establishing biofabrication approaches as clinically relevant tools. Achieving this requires bio-inks which must not only be easily printable, but also allow controllable and reproducible printing of cells. This review outlines the general principles and current progress and compares the advantages and challenges for the most widely used biofabrication techniques for printing cells: extrusion, laser, microvalve, inkjet and tissue fragment printing. It is expected that significant advances in cell printing will result from synergistic combinations of these techniques and lead to optimised resolution, throughput and the overall complexity of printed constructs.


Three residues in the common beta chain of the human GM-CSF, IL-3 and IL-5 receptors are essential for GM-CSF and IL-5 but not IL-3 high affinity binding and interact with Glu21 of GM-CSF.
The beta subunit (beta c) of the receptors for human granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-3 (IL-3) and interleukin-5 (IL-5) is essential for high affinity ligand-binding and signal transduction. An important feature of this subunit is its common nature, being able to interact with GM-CSF, IL-3 and IL-5. Analogous common subunits have also been identified in other receptor systems including gp130 and the IL-2 receptor gamma subunit. It is not clear how common receptor subunits bind multiple ligands. We have used site-directed mutagenesis and binding assays with radiolabelled GM-CSF, IL-3 and IL-5 to identify residues in the beta c subunit involved in affinity conversion for each ligand. Alanine substitutions in the region Tyr365-Ile368 in beta c showed that Tyr365, His367 and Ile368 were required for GM-CSF and IL-5 high affinity binding, whereas Glu366 was unimportant. In contrast, alanine substitutions of these residues only marginally reduced the conversion of IL-3 binding to high affinity by beta c. To identify likely contact points in GM-CSF involved in binding to the 365-368 beta c region we used the GM-CSF mutant eco E21R which is unable to interact with wild-type beta c whilst retaining full GM-CSF receptor alpha chain binding. Eco E21R exhibited greater binding affinity to receptor alpha beta complexes composed of mutant beta chains Y365A, H367A and I368A than to those composed of wild-type beta c or mutant E366A. These results (i) identify the residues Tyr365, His367 and Ile368 as critical for affinity conversion by beta c, (ii) show that high affinity binding of GM-CSF and IL-5 can be dissociated from IL-3 and (iii) suggest that Tyr365, His367 and Ile368 in beta c interact with Glu21 of GM-CSF.


Sports related to drowning.
Aquatic sports are included in the top list of risky practices as the environment per se carries a possibility of death by drowning if not rescued in time. Not only are aquatic sports related to a high risk of death, but also all sports practiced on the water, over the water and on ice. Whatever the reason a person is in the water, drowning carries a higher possibility of death if the individual is unable to cope with the water situation, which may simply be caused by an inability to stay afloat and get out of the water or by an injury or disease that may lead to physical inability or unconsciousness. The competitive nature of sports is a common pathway that leads the sports person to exceed their ability to cope with the environment or simply misjudge their physical capability. Drowning involves some principles and medical interventions that are rarely found in other medical situations as it occurs in a deceptively hostile environment that may not seem dangerous. Therefore, it is essential that health professionals are aware of the complete sequence of action in drowning. This article focuses on the pulmonary injury in sports and recreational activities where drowning plays the major role.


Cloning of alpha 1(IV) and alpha 2(IV) collagen cDNAs from rabbit corneal endothelial cell RNA.
To understand the role of type IV collagen in embryogenesis, regeneration, and tissue repair of the cornea at the molecular level, the authors isolated clones coding for rabbit alpha 1(IV) and alpha 2(IV) chains from a cDNA library constructed with rabbit corneal endothelial cell RNA. The isolated alpha 2(IV) cDNA clones encode a part of a 5' untranslated region, the signal peptide, the 7S domain, part of the triple-helical domain, and the entire carboxyl-terminal nontriple-helical (NC1) domain. By cross hybridization, using one of the alpha 2(IV) cDNA inserts, a cDNA clone encoding the alpha 1(IV) chain was isolated from a lambda gt10 library primed with oligo(dT). The clone covered a short portion of the triple-helical domain, the entire NC1 domain, and a short 3' untranslated region. The nucleotide-sequence analysis of these clones provides, for the first time to the authors' knowledge, the primary structure of the carboxyl-terminal portion of both rabbit alpha 1(IV) and alpha 2(IV) collagen chains. Between the alpha 1(IV)NC1 and the alpha 2(IV)NC1, 61% and 66% sequence similarities were observed at the amino acid and nucleotide levels, respectively. The locations of all the 12 cysteinyl residues were conserved in the two NC1 sequences.


Endocrine effects of relaxin overexpression in mice.
Relaxin is a small peptide hormone with a variety of biological functions. To investigate the systemic endocrine effects of relaxin, we produced mice with transgenic overexpression of the Rln1 gene, Tg(Rln1), driven by rat insulin 2 promoter. The expression of relaxin was detected in the pancreas of the transgenic animals. An analysis of the sera from the transgenic animals revealed at least 20-fold elevation of the level of bioactive relaxin. Transgenic animals had normal viability and fertility in both sexes. Transgenic overexpression of Rln1 did not rescue the undescended testis phenotype in Insl3-deficient males, suggesting that in vivo relaxin does not interact with the insulin-like 3 factor receptor, leucine-rich repeats-containing G protein-coupled receptor 8, Lgr8. Phenotypically, the excess of relaxin resulted in hypertrophic nipple development in virgin female mice. Deletion of the relaxin receptor, leucine-rich repeats-containing G protein-coupled receptor 7, Lgr7, in Tg(Rln1) animals abrogated the development of enlarged nipples in females, indicating that relaxin exerts its effect through Lgr7 alone. The levels of previously defined targets of relaxin signaling, such as matrix metalloproteinases 2 and 9, vascular endothelial growth factor, or nitric oxide, were similar in the sera of the transgenic and wild-type mice. However, the total plasma protein concentration in male Tg(Rln1) mice was lower than that in control animals. The livers of male Tg(Rln1) mice exhibited significantly higher hydroxyproline content, indicative of increased collagen deposition. Our results indicate that relaxin overexpression causes gender-specific changes in liver collagen metabolism.