Optimal deployment of attentional gain during fine discriminations.
Most models assume that top-down attention enhances the gain of sensory neurons tuned to behaviorally relevant stimuli (on-target gain). However, theoretical work suggests that when targets and distracters are highly similar, attention should enhance the gain of neurons that are tuned away from the target, because these neurons better discriminate neighboring features (off-target gain). While it is established that off-target neurons support difficult fine discriminations, it is unclear if top-down attentional gain can be optimally applied to informative off-target sensory neurons or if gain is always applied to on-target neurons, regardless of task demands. To test the optimality of attentional gain in human visual cortex, we used functional magnetic resonance imaging and an encoding model to estimate the response profile across a set of hypothetical orientation-selective channels during a difficult discrimination task. The results suggest that top-down attention can adaptively modulate off-target neural populations, but only when the discriminanda are precisely specified in advance. Furthermore, logistic regression revealed that activation levels in off-target orientation channels predicted behavioral accuracy on a trial-by-trial basis. Overall, these data suggest that attention does not only increase the gain of sensory-evoked responses, but may bias population response profiles in an optimal manner that respects both the tuning properties of sensory neurons and the physical characteristics of the stimulus array.


Tetrahydrobiopterin recycling, a key determinant of endothelial nitric-oxide synthase-dependent signaling pathways in cultured vascular endothelial cells.
Tetrahydrobiopterin (BH4) is a key redox-active cofactor in endothelial isoform of NO synthase (eNOS) catalysis and is an important determinant of NO-dependent signaling pathways. BH4 oxidation is observed in vascular cells in the setting of the oxidative stress associated with diabetes. However, the relative roles of de novo BH4 synthesis and BH4 redox recycling in the regulation of eNOS bioactivity remain incompletely defined. We used small interference RNA (siRNA)-mediated "knockdown" GTP cyclohydrolase-1 (GTPCH1), the rate-limiting enzyme in BH4 biosynthesis, and dihydrofolate reductase (DHFR), an enzyme-recycling oxidized BH4 (7,8-dihydrobiopterin (BH2)), and studied the effects on eNOS regulation and biopterin metabolism in cultured aortic endothelial cells. Knockdown of either DHFR or GTPCH1 attenuated vascular endothelial growth factor (VEGF)-induced eNOS activity and NO production; these effects were recovered by supplementation with BH4. In contrast, supplementation with BH2 abolished VEGF-induced NO production. DHFR but not GTPCH1 knockdown increased reactive oxygen species (ROS) production. The increase in ROS production seen with siRNA-mediated DHFR knockdown was abolished either by simultaneous siRNA-mediated knockdown of eNOS or by supplementing with BH4. In contrast, addition of BH2 increased ROS production; this effect of BH2 was blocked by BH4 supplementation. DHFR but not GTPCH1 knockdown inhibited VEGF-induced dephosphorylation of eNOS at the inhibitory site serine 116; these effects were recovered by supplementation with BH4. These studies demonstrate a striking contrast in the pattern of eNOS regulation seen by the selective modulation of BH4 salvage/reduction versus de novo BH4 synthetic pathways. Our findings suggest that the depletion of BH4 is not sufficient to perturb NO signaling, but rather that concentration of intracellular BH2, as well as the relative concentrations of BH4 and BH2, together play a determining role in the redox regulation of eNOS-modulated endothelial responses.


Neutrophils are required for both the sensitization and elicitation phase of contact hypersensitivity.
Allergic contact dermatitis and its animal model, contact hypersensitivity (CHS), are T cell-mediated inflammatory skin diseases induced by contact allergens. Though numerous cellular and molecular players are known, the mechanism of chemical-induced sensitization remains poorly understood. Here, we identify neutrophils as crucial players in the sensitization phase of CHS. Genetic deficiency of neutrophils caused by myeloid-specific deletion of Mcl-1 or antibody-mediated depletion of neutrophils before sensitization abrogated the CHS response. Neutrophil deficiency reduced contact allergen-induced cytokine production, gelatinase release, and reactive oxygen species production in naive mice. Mast cell deficiency inhibited neutrophil accumulation at the site of sensitization. In turn, neutrophils were required for contact allergen-induced release of further neutrophil-attracting chemokines, migration of DCs to the draining lymph nodes, and priming of allergen-specific T cells. Lymph node cells from mice sensitized in the absence of neutrophils failed to transfer sensitization to naive recipients. Furthermore, no CHS response could be induced when neutrophils were depleted before elicitation or when normally sensitized lymph node cells were transferred to neutrophil-deficient recipients, indicating an additional role for neutrophils in the elicitation phase. Collectively, our data identify neutrophils to be critically involved in both the sensitization and elicitation phase of CHS.


Prostate-specific antigen production in the female breast: association with progesterone.
Prostate-specific antigen (PSA) is produced by the female breast. Prior in vitro evidence suggests that PSA expression in breast epithelial cells is regulated by androgens and progestins but not estrogens. The purpose of this study was to determine whether (a) PSA expression in breast nipple aspirate fluid (NAF) and in serum is influenced by progesterone (PG); (b) the ability to obtain NAF decreases with repeated breast aspirations; and (c) PSA in NAF correlates with abnormal NAF cytology. Eight pre- and three postmenopausal women with no breast cancer risk factors were enrolled in a pilot study and had NAF and serum collected every 3-4 days for a month to evaluate the influence of serum PG, luteinizing hormone, estradiol, and follicle-stimulating hormone on PSA in serum and in NAF. NAF was obtained in 99% (112 of 113) of aspiration visits. Median, mean, and peak NAF but not serum PSA levels were higher in pre- than in postmenopausal subjects. NAF PSA levels were associated with the rise or peak in serum PG in seven of eight premenopausal women (seven of seven with a PG surge) and in zero of three postmenopausal women. Considering all 11 women, there was an association between NAF PSA and PG (P = 0.005) but not luteinizing hormone, estradiol, or follicle-stimulating hormone. NAF volume did not significantly change over time. Atypical hyperplasia (9%) and hyperplasia without atypia (36%) were identified in the NAF of a subset of the subjects. Median, mean, and peak levels of NAF PSA (P = 0.05, 0.05, and 0.10, respectively) were higher in subjects with normal versus hyperplastic cytology. PSA production in the breast increases in association with PG. With aspiration every 3-4 days, NAF volume does not significantly decrease over time. NAF cytology and PSA levels in NAF may help identify women at increased breast cancer risk. Changes in biomarkers of breast cancer risk in NAF (including PSA and cytology) may predate mammographic abnormalities. NAF may, therefore, be useful as a breast cancer screening tool for young women who are not recommended to undergo mammography and as an adjunct to screen women who have mammograms performed.


Single nucleotide polymorphism in the microRNA-199a binding site of HIF1A gene is associated with pancreatic ductal adenocarcinoma risk and worse clinical outcomes.
Hypoxia-inducible factor-1 alpha (HIF-1α) is over-expressed in many cancers including pancreatic ductal adenocarcinoma (PDAC) and correlated with poor prognosis. We aim to determine the effect of germline genetic variants on the regulation of the homeostasis of the miRNA-gene regulatory loop in HIF1A gene and PDAC risk. HIF1A rs2057482 single nucleotide polymorphism (SNP) was genotyped in 410 PDAC cases and 490 healthy controls. The CC genotype SNP HIF1A is significantly correlated with PDAC risk (OR = 1.719, 95% CI: 1.293-2.286) and shorter overall survival (OS, P<0.0001) compared with the CT/TT alleles group. The C/T variants of rs2057482, a SNP located near the miR-199a binding site in HIF1A, could lead to differential regulation of HIF1A by miR-199a. Specifically, the C allele of rs2057482 weakened miR-199a-induced repression of HIF-1α expression on both mRNA and protein levels. In the PDAC tissue, individuals with the rs2057482-CC genotype expressed significantly higher levels of HIF-1α protein than those with the rs2057482-CT/TT genotype (P<0.0001). Both the CC genotype of SNP HIF1A and increased HIF-1α expression are significantly associated with shorter OS of patients with PDAC. After adjusted by TNM staging, differentiation grade, and the levels of CA19-9, both SNP HIF1A and HIF-1α expression retained highly significance on OS (P<0.0001). Taken together, our study demonstrates that host genetic variants could disturb the regulation of the miR-199a/HIF1A regulatory loop and alter PDAC risk and poor prognosis. In conclusion, the rs2057482-CC genotype increases the susceptibility to PDAC and associated with cancer progression.


Dietary intakes of retinol, carotenes, vitamin C, and vitamin E and colorectal cancer risk: the Fukuoka colorectal cancer study.
It has long been a matter of interest whether antioxidant vitamins are protective against colorectal cancer as well as human cancers in general, but epidemiological evidence is inconclusive. We investigated associations of dietary intakes of retinol and antioxidant vitamins with colorectal cancer risk in 816 incident cases of histologically confirmed colorectal cancer and 815 controls randomly selected for the Fukuoka colorectal cancer study in Japan. Dietary intakes were assessed by a PC-assisted interview regarding 148 food items. Statistical adjustment was made for body mass index, physical activity, calcium, and n-3 fatty acid intake and other factors. Retinol intake was significantly, inversely associated with colorectal cancer risk; the odds ratio for the highest vs. lowest was 0.55 (95% CI: 0.35, 0.88; P (trend) = 0.01) in women, but a modest increase in the risk was observed among men with the highest intake of retinol. Liver was the major source of retinol intake and showed similar associations with colorectal cancer risk in men and women. Intake of carotenes, vitamin C, and vitamin E were not related to colorectal cancer risk in either men or women. The study did not support a hypothesis that dietary intake of antioxidant vitamins is protective in the development of colorectal cancer.


Long-term survival in patients with mild or moderate impairment of left ventricular contractility during routine diagnostic left ventriculography.
Long-term survival in patients with mild to moderate impairment of left ventricular contractility in the absence of coronary artery disease has not been studied extensively but the prognosis is assumed to be good. One hundred sixty-eight patients with angiographic evidence of mild or moderate impairment of left ventricular contractility and no other significant cardiac disease at the time of routine diagnostic cardiac catheterization were studied to determine long-term survival and event-free survival. Clinical characteristics, electrocardiograms, chest X-rays, laboratory data, and hemodynamics including end-diastolic volume, end-systolic volume, stroke volume, ejection fraction, and regional wall motion at the time of catheterization were examined. Patients with moderate impairment were more likely to have dyspnea (P = 0.005) and an abnormal electrocardiogram (P = 0.006) than patients with mild impairment. Mean ejection fraction was 57% (P = 0.0001 vs. normal) in patients with mild impairment and was 47% (P = 0.0001 vs. normal) in patients with moderate impairment. Wall motion studies showed impairment to be generalized and more significantly abnormal in patients with moderate impairment. Survival at a mean of 138 months could be determined in 162 of the 168 patients (96%). Fourteen year actuarial survival was 92% for patients with mild impairment compared to 75% for patients with moderate impairment (P = 0.01). Long-term prognosis is good in patients found to have mild generalized impairment at the time of routine diagnostic cardiac catheterization. In patients with moderate impairment, closer follow-up to prevent arrhythmia and the onset of congestive heart failure appears to be warranted.


Mollaret's meningitis. A variant of recurrent hereditary polyserositis, both provoked by metaraminol.
Mollaret's meningitis is a rare condition with a characteristic clinical and cerebrospinal fluid picture. In many ways it resembles recurrent hereditary polyserositis (familial Mediterranean fever) in its natural history, pattern of attacks, and response to colchicine. Association of the two conditions has been reported, so far, in two patients only. In our patient the symptoms of both conditions were induced by a metaraminol provocative infusion. We have previously introduced this as a specific diagnostic and confirmatory test for recurrent hereditary polyserositis. The possibility that the two conditions represent different manifestations of a single disease is therefore strengthened.


A prospective longitudinal study of seasonality in African students living in the greater Washington, D.C. metropolitan area.
We conducted a prospective, longitudinal study of seasonality in a vulnerable population, i.e., African students who migrated to a temperate climate. Consistent with previous cross-sectional studies, we hypothesized lower mood and energy, and higher appetite and weight, in fall/winter than in spring/summer. Four cohorts of African students attending a year-long nursing school program without vacation in Washington, D.C., were assessed monthly for 1 year. Forty-three subjects (mean age = 33.46 +/- 6.25), consisting of predominantly females (76.7%), completed the study. The cohorts began their academic program in different seasons (one each in winter, spring, summer, and fall), inherently minimizing confounding influences on seasonality, such as academic and immigration stress, as well as allowing adjustment for an order effect. At each assessment, students completed three 100-mm visual analog scales for mood, energy, and appetite, and were weighed on a digital scale. For each standardized dependent variable, a repeated measure ANOVA was used and, if a significant effect of month was identified, averages for spring/summer and fall/winter were compared using paired t-tests. In addition, a mixed model for repeated measures was applied to raw (nonstandardized) data. Body weight was significantly higher in fall/winter than in spring/summer (p < 0.01). No seasonal differences in mood, energy, or appetite were found. Benefiting from certain unique features of our cohorts allowing adjustment for order effects, this is the first study to identify a seasonal variation in body weight with a peak in winter using longitudinal monthly measurements.


Effect of IL-1beta, PGE(2), and TGF-beta1 on the expression of OPG and RANKL in normal and osteoporotic primary human osteoblasts.
The RANKL/RANK/OPG pathway is essential for bone remodeling regulation. Many hormones and cytokines are involved in regulating gene expression in most of the pathway components. Moreover, any deregulation of this pathway can alter bone metabolism, resulting in loss or gain of bone mass. Whether osteoblasts from osteoporotic and nonosteoporotic patients respond differently to cytokines is unknown. The aim of this study was to compare the effect of interleukin (IL)-1beta, proftaglandin E(2) (PGE(2)), and transforming growth factor-beta1 (TGF-beta1) treatments on OPG and RANKL gene expression in normal (n = 11) and osteoporotic (n = 8) primary osteoblasts. OPG and RANKL mRNA levels of primary human osteoblastic (hOB) cell cultures were assessed by real-time PCR. In all cultures, OPG mRNA increased significantly in response to IL-1beta treatment and decreased in response to TGF-beta1 whereas PGE(2) treatment had no effect. RANKL mRNA levels were significantly increased by all treatments. Differences in OPG and RANKL responses were observed between osteoporotic and nonosteoporotic hOB: in osteoporotic hOB, the OPG response to IL-1beta treatment was up to three times lower (P = 0.009), whereas that of RANKL response to TGF-beta1 was five times higher (P = 0.002) after 8 h of treatment, as compared with those in nonosteoporotic hOBs. In conclusion, osteoporotic hOB cells showed an anomalous response under cytokine stimulation, consistent with an enhanced osteoclastogenesis resulting in high levels of bone resorption.


No effect of intrarenal converting enzyme inhibition on canine renal blood flow.
Captopril and teprotide were administered intra-arterially to the kidney and intravenously to inhibit intrarenal and extrarenal converting enzyme (CE), respectively. Captopril was infused at 0.4, 0.8, and 1.6 micrograms . kg-1 . min-1 intra-arterially, and teprotide was given at 0.4 and 0.8 micrograms . kg-1 . min-1 intra-arterially in salt-replete dogs (group 1). Both agents were also given intravenously in the dose of 0.2 mg/kg, known to cause maximal extrarenal CE inhibition. The intra-arterial infusions of the inhibitors had no effect on renal hemodynamics but caused a graded degree of intrarenal CE inhibition. A lesser degree of extrarenal CE inhibition was seen after captopril in these doses. Intravenous administration of captopril and teprotide inhibited extrarenal CE, but only captopril increased renal blood flow (13%). When teprotide was given in a higher dose (group 3, 1.02 mg/kg), it too increased renal blood flow. In salt-deplete dogs (group 2), captopril infused intra-arterially caused a degree of intrarenal CE inhibition comparable with that in the salt-replete dogs but again produced little or no renal hemodynamic changes. When given intravenously in this group, captopril inhibited extrarenal EE and elicited a greater increase in renal blood flow (36%) than in the group 1 dogs. These results indicate that in conscious dogs renal vasodilatation is associated with extrarenal, but not intrarenal, CE inhibition.


Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal.
The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium-an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5+ intestinal stem cells (ISCs). R-spondin ligands (RSPO1-RSPO4) engage distinct LGR4-LGR6, RNF43 and ZNRF3 receptor classes, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5+ ISCs in vivo. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5+ ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5+ ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration.


Cannabinoid exposure during pregnancy and its impact on immune function.
Cannabinoids are the most commonly abused illicit drugs worldwide. While cannabis can be beneficial for certain heath conditions, abuse of potent synthetic cannabinoids has been on the rise. Exposure to cannabinoids is also prevalent in women of child-bearing age and pregnant women. These compounds can cross the placental barrier and directly affect the fetus. They mediate their effects primarily through G-protein coupled cannabinoid receptors, CB1 and CB2. In addition to significant neurological effects, cannabinoids can trigger robust immunomodulation by altering cytokine levels, causing apoptosis of lymphoid cells and inducing suppressor cells of the immune system. Profound effects of cannabinoids on the immune system as discussed in this review, suggest that maternal exposure during pregnancy could lead to dysregulation of innate and adaptive immune system of developing fetus and offspring potentially leading to weakening of immune defenses against infections and cancer later in life. Emerging evidence also indicates the underlying role of epigenetic mechanisms causing long-lasting impact following cannabinoid exposure in utero.


Net basolateral potassium flux and short-circuit current in ouabain-treated frog skin.
A new technique has been developed to correlate K loss from cells (JK) across the basolateral membrane into a K-free ouabain Ringer solution and short-circuit current (Isc) for a model Na-transporting epithelium, the frog skin. Distinct differences were observed when the tissue was bathed in sulfate or chloride Ringer. In sulfate Ringer, K-free ouabain treatment caused both JK and Isc to decline in a nearly parallel fashion with time. JK-Isc was approximately 1 microA/cm2. In sulfate Ringer, isoproterenol caused parallel increases, whereas amiloride (apical side) caused parallel decreases in JK and Isc. In chloride Ringer, K-free ouabain treatment caused Isc to decline at a slightly faster rate than JK.JK-Isc was approximately 8 microA/cm2. Bumetanide decreased JK with very little effect on Isc. Barium caused small parallel changes in both Isc and JK. Amiloride decreased Isc with very little effect on JK. These experiments show that after ouabain treatment changes in JK from the cells across the basolateral membrane can largely account for changes in Isc. However, JK also occurs via neutral mechanisms and perhaps from cells not related to the transport pathway, demonstrating that there is not always a tight coupling of K loss at the basolateral membrane with Na entry across the apical membrane.


Type IV collagenases in human amniotic fluids and amnion epithelial cells.
Gelatin zymograms revealed in human second trimester amniotic fluids a constant M(r) 66,000 proteolytically active polypeptide that in immunoblotting was recognized by antiserum against the human fibroblast gelatinase/type IV collagenase. Isolated cultured human amnion epithelial cells produced the M(r) 66,000 proteinase and frequently a M(r) 92,000 one. The production of the former was enhanced by transforming growth factor-beta and that of the latter by tetradecanoyl phorbol acetate. Immunoprecipitation experiments indicated that the M(r) 66,000 amnion epithelial cell proteinase was identical with that found in the amniotic fluids and that the M(r) 92,000 one corresponded to the human macrophage/granulocyte gelatinase/type IV collagenase. Approximate quantifications of the M(r) 66,000 proteinase in amniotic fluids and sera from same individuals were assessed by comparison of dilution series of the samples and standard enzyme preparations in zymography. These measurements indicated that the amount of the proteinase in the amniotic fluids was about 5-10 micrograms/ml representing 0.1-0.2% of total proteins. In sera the corresponding figures were 30-40 micrograms/ml and 0.03-0.06%, respectively. Thus the relative concentration of the M(r) 66,000 proteinase in amniotic fluids was approx. 3-4-fold that in sera, suggesting local production of the protein. The proteinase may be produced by amnion epithelial cells that under so far unknown physiologic stimulus also may start secreting the M(r) 92,000 enzyme.


Characteristics of very poor outcome schizophrenia.
The authors compared 21 "Kraepelinian" schizophrenic patients who had been ill and dependent on others for the past 5 years with 76 chronic schizophrenic patients in remission or with exacerbations requiring hospitalization. The Kraepelinian patients met the criteria for schizophrenia by more diagnostic systems than the exacerbated patients, were less responsive to haloperidol, had more severe negative symptoms, and had similarly severe positive symptoms. They had cerebral ventricles that were more asymmetrical and a greater family history of schizophrenia spectrum disorders than the other chronic patients. These data suggest that patients with 5 years of illness and complete dependency on others may represent a subgroup of schizophrenia.


Parasitic nematode-induced modulation of body weight and associated metabolic dysfunction in mouse models of obesity.
Obesity is associated with a chronic low-grade inflammation characterized by increased levels of proinflammatory cytokines that are implicated in disrupted metabolic homeostasis. Parasitic nematode infection induces a polarized Th2 cytokine response and has been explored to treat autoimmune diseases. We investigated the effects of nematode infection against obesity and the associated metabolic dysfunction. Infection of RIP2-Opa1KO mice or C57BL/6 mice fed a high-fat diet (HFD) with Nippostrongylus brasiliensis decreased weight gain and was associated with improved glucose metabolism. Infection of obese mice fed the HFD reduced body weight and adipose tissue mass, ameliorated hepatic steatosis associated with a decreased expression of key lipogenic enzymes/mediators, and improved glucose metabolism, accompanied by changes in the profile of metabolic hormones. The infection resulted in a phenotypic change in adipose tissue macrophages that was characterized by upregulation of alternative activation markers. Interleukin-13 (IL-13) activation of the STAT6 signaling pathway was required for the infection-induced attenuation of steatosis but not for improved glucose metabolism, whereas weight loss was attributed to both IL-13/STAT6-dependent and -independent mechanisms. Parasitic nematode infection has both preventive and therapeutic effects against the development of obesity and associated features of metabolic dysfunction in mice.


Up-regulation of NOD1 and NOD2 through TLR4 and TNF-alpha in LPS-treated murine macrophages.
NOD1 (Card4) and NOD2 (Card15) are thought to be responsible for cytoplasmic defense against bacterial entry. To gain further knowledge about how their expressions are regulated in murine macrophages, we investigated the expression of NOD1 and NOD2 mRNAs after stimulation with various endotoxins, lipopolysaccharide, lipoteichoic acid and peptidoglycan. In macrophage RAW264.7 cells, the first and second rises in NOD1 and NOD2 mRNAs were observed at 2 hr and at 8-12 hr after endotoxin treatment. Increases in NOD1 and NOD2 mRNAs at 2 hr in lipopolysaccharide-treated RAW264.7 cells were reduced with the use of NF-kappaB inhibitor, caffeic acid phenethyl ester. In RAW264.7 cells, lipopolysaccharide-induced increases in NOD1 and NOD2 mRNAs were inhibited with anti-TLR4 antibody, and partially reduced in peritoneal macrophages obtained from TLR4-deficient mice. Furthermore, NOD1 and NOD2 mRNA expressions in RAW264.7 cells were increased by the treatment with proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), or IL-6. In TNF-alpha deficient macrophages, the expression of NOD molecules was minimal at 12 hr, and the second rise in NOD mRNA seen in lipopolysaccharide-treated RAW264.7 cells was inhibited with anti-TNF-alpha, but not with anti-IL-1beta or anti-IL-6 antibody. These observations suggest that immediate response of NODs to endotoxins could result from NF-kappaB activation via TLR signaling, whereas the second rise in NOD mRNAs might have resulted from TNF-alpha production possibly through NF-kappaB, TLR, and/or NOD signalings.


Walk Texas! 5-A-Day intervention for Women, Infant, and Children (WIC) clients: a quasi-experimental study.
This study was conducted to evaluate the Walk Texas! Clinical Counseling Guide for Nutrition, which is a brief, stage-based nutritional counseling guide designed for use in clinical settings. This study utilized a pre-test post-test quasi-experimental design, with two intervention and two comparison clinics that were matched for size and ethnicity. Intervention participants were staged for readiness to meet the 5-A-Day criteria for fruits and vegetables (F&V) and provided stage-based counseling. Women, Infant, and Children (WIC) counselors were trained to utilize the Walk Texas! brief staging instrument and to provide all counseling. Primary measures included: stage of change, barriers to change, attitudes toward F&V, self-efficacy, and self-reports of F&V consumption. These were collected at baseline and during follow-up visits to the WIC clinic. A total of 433 participants completed pre-test data across the four sites. A number of individuals transferred from their original clinic or did not return to their clinic during the study (n=62). Of the remaining 371 participants, a total of 225 participants returned at least two surveys, for a response rate of 60.6%. These were primarily native Spanish speaking, Hispanic women, of low educational level. Results were mixed. Although there was no significant increase in the cognitive constructs (self-efficacy, attitudes, etc.), participants in the intervention clinics reported a significant increase in stage of change and a composite measure of F&V in-take. In contrast, participants in the control clinics reported no change in these variables. Thus, there appears to be some utility in the use of the Walk Texas! Clinical Counseling Guide for Nutrition in this population.


Neonatal posterior fossa haemorrhage: a difficult ultrasound diagnosis.
Four newborn neonates with a history of birth trauma and significant neurological signs were examined with both ultrasound and computed tomography (CT) performed within hours of each other. Ultrasound was inadequate in initial assessment, as it missed or underestimated posterior fossa haemorrhage, particularly where this was extraparenchymal or vermian. CT is the examination of choice in these patients.


Swedish School Nurses' Experiences of Child Abuse.
Child abuse recognition and the protection of children is a global concern. In Sweden, the role of the school nurse (SN) is to promote schoolchildren's health and development and to identify and prevent harm. The aim of this study was to describe Swedish SN experiences of suspecting, identifying, and reporting child abuse and to compare them with respect to (a) years of experience as SN, (b) age of SN, and (c) pupil population size. A descriptive design was used. Two-hundred and thirty-three SNs completed a survey detailing their experiences. Most SNs (96%) reported having suspected a child suffering from physical or psychological abuse. Approximately half of them reported occurrences of honor-related violence (54%) and of child sexual abuse (57%). SNs with less nursing experience reported significantly less recognition and reporting of child abuse. The findings indicate that experiences of child abuse are common. Thus, it is vital that SNs have the necessary competency and support to identify and report suspected child abuse.


Monoclonal antibodies for the measurement of class-specific antibody responses in the green turtle, Chelonia mydas.
Monoclonal antibodies (Mabs) were developed against the known immunoglobulin classes of the green turtle, Chelonia mydas. Plasma protein fractions enriched for 5.7S IgY, 7S IgY, and IgM turtle immunoglobulins were used to immunize Balb/c mice for hybridoma production and for hybridoma screening. Fifteen hybridomas produced Mabs with specificity for turtle immunoglobulins and for affinity purified dinitrophenol (DNP) specific turtle antibodies. Three Mabs specific for either turtle 5.7S IgY heavy chain (HL814), 7S IgY heavy chain (HL857), or IgM heavy chain (HL846) were purified and used in an enzyme-linked immunosorbent assay (ELISA) to measure antibody responses in two turtles immunized with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) over a 10 month period. In both turtles the 7S IgY antibody response developed within 5 weeks of the first inoculation and remained high over the following 9 months. The 5.7S IgY antibody response was detected in one turtle at 3-4 months and in the other at 8 months, and reached high levels in both individuals by 10 months. The IgM responses were difficult to interpret. One turtle had pre-inoculation anti-DNP IgM antibody in its plasma and the other developed only a weak, transient response at about 4 months. The class-specific antibody activity in immune turtle plasma could be strongly inhibited by soluble DNP or by rabbit anti-DNP specific antiserum, showing that these antibody responses were directed predominantly to the DNP hapten on the DNP-BSA antigen. Antibody responses to the BSA carrier could not be detected in either turtle over the course of the immunization. Mab HL814, specific for an epitope on the 5.7S green turtle immunoglobulin heavy chain, will be useful for characterizing the molecular relationships of 5.7S, 7S and IgM heavy chains and the role of 5.7S antibody in humoral immunity in this species. All anti-turtle Ig Mabs were screened against the plasma globulins of Loggerhead (Caretta caretta), Olive Ridley (Lepidochelys olivacea), Kemp's Ridley (Lepidochelys kempi), Hawksbill (Eretmochelys imbricata), and Leatherback (Dermochelys coriacea). While the Mabs specific for IgM and 5.7S IgY reacted only with the green turtle, two Mabs specific for light chain reacted with all species except the leatherback, and nine mabs specific for 7S IgY heavy chain reacted with all five species. Thus, these Mabs may be useful for immunodiagnostic applications in these endangered species as well.


BRAF V600E mutation in metastatic colorectal cancer: Methods of detection and correlation with clinical and pathologic features.
The screening for BRAF V600E mutation is employed in clinical practice for its prognostic and potentially predictive role in patients with metastatic colorectal carcinoma (mCRC). Little information is available on the sensitivity and specificity of the testing methods to detect this mutation in CRC. By using serial dilution of BRAF mutant DNA with wild type DNA, we found that the sensitivity of allelic discrimination-Real Time PCR was higher than PCR-Sequencing (10% vs 20%). In agreement, the Real Time PCR assay displayed increased analytical sensitivity in detecting the BRAF V600E mutation as compared with PCR-Sequencing in a cohort of 510 consecutive CRCs (21 vs 16 cases). Targeted resequencing demonstrated that all cases negative by PCR-Sequencing had an allelic frequency of the BRAF mutation <20%, thus suggesting tumor heterogeneity. The association of BRAF mutations with clinical and pathological features was assessed next in a cohort of 840 KRAS exon 2 wild type CRC patients screened with the Real Time PCR assay. The BRAF V600E mutation frequency in this cohort was 7.8% that increased to 33.4% in females over 70 y of age with right-sided tumor location. BRAF mutations were also detected in 4.4% of male patients with left-sided tumors and aged <70 y. Fourteen of 61 (22.9%) BRAF V600E mutation bearing patients exhibited microsatellite instability (MSI) as assessed by T17 mononucleotide sequence within intron 8 of HSP110. Our study indicates that Real Time PCR-based assays are more sensitive than PCR-Sequencing to detect the BRAF V600E mutation in CRC and that BRAF mutations screening should not be restricted to selected patients on the basis of the clinical-pathological characteristics.


The role of ozone flux and antioxidants in the suppression of ozone injury by elevated CO2 in soybean.
The projected rise in atmospheric CO2 concentration is expected to increase growth and yield of many agricultural crops. The magnitude of this stimulus will partly depend on interactions with other components of the atmosphere such as tropospheric O3. Elevated CO2 concentrations often lessen the deleterious effects of O3, but the mechanisms responsible for this response have received little direct examination. Previous studies have indicated that protection against O3 injury by elevated CO2 can be attributed to reduced O3 uptake, while other studies suggest that CO2 effects on anti-oxidant metabolism might also be involved. The aim of this experiment was to test further the roles of O3 flux and antioxidant metabolism in the suppression of O3 injury by elevated CO2. In a two-year experiment, soybean [Glycine max (L.) Merr.] was exposed from emergence to maturity to charcoal-filtered air or charcoal-filtered air plus a range of O3 concentrations in combination with ambient or approximately twice-ambient CO2 concentrations in open-top field chambers. Experimental manipulation of O3 concentrations and estimates of plant O3 uptake indicated that equivalent O3 fluxes that suppressed net photosynthesis, growth, and yield at ambient concentrations of CO2 were generally much less detrimental to plants treated concurrently with elevated CO2. These responses appeared unrelated to treatment effects on superoxide dismutase, glutathione reductase, and peroxidase activities and glutathione concentration. Total ascorbic acid concentration increased by 28-72% in lower canopy leaves in response to elevated CO2 and O3 but not in upper canopy leaves. Increasing concentrations of atmospheric CO2 will likely ameliorate O3 damage to many crops due to reduced O3 uptake, increased carbon assimilation, and possibly as yet undetermined additional factors. The results of this study further suggest that elevated CO2 may increase the threshold O3 flux for biomass and yield loss in soybean.


Childhood and current cognitive function in healthy 80-year-olds: a DT-MRI study.
Diffusion tensor MR imaging (DT-MRI) yields information on early pathological changes in white matter of the ageing brain which may correlate with cognitive function. However, because individuals vary in their cognitive ability, a measurement of prior cognition from youth is required to understand fully the significance of MR imaging changes associated with ageing. Here, diffusion tensor parameters and cognitive function were measured in a cohort of 30 older subjects whose cognitive ability was measured at age 11 and 80. There was a significant correlation between diffusion anisotropy measured in the centrum semiovale at age 80 and mental ability determined at age 11 and 80. These novel results suggest, that MR imaging studies of white matter structure and its relationship to mental ability in ageing should control for early life cognition.


Computational-based identification and analysis of globally expressed differential genes in high-grade serous ovarian carcinoma cell lines.
Ovarian Cancer (OVCA) is the most occurring gynecological cancer worldwide, often diagnosed at a later stage and ultimate results in a high death rate. To overcome this serious health concern, it is important to understand the molecular mechanisms and equally significant to identify the putative biomarkers as well as the therapeutic drug targets for the early diagnosis and treatment of OVCA. In doing so, a strategy is designed to study the most frequently diagnosed cases of OVCA called as High-Grade Serous Ovarian Carcinoma (HGSOC) cell lines with the combination of computational biology, biostatistics and cancer informatics approaches. This study is directed to investigate the global gene expression profiling, and to perform the analyses of identified global Differently Expressed Genes (DEGs) of OVCA. The microarray dataset (GSE71524) is comprised of tumor and cell line samples of OVCA and it was used for the identification of DEGs in the current study. The STRING database was used to construct Protein-Protein Interaction (PPI) network of DEGs, and hub genes were identified by the CytoHubba. In addition, a functional enrichment analysis of up- and down-regulated DEGs was performed by a bioinformatics database called as DAVID. The microRNAs (miRNAs) and transcription factors (TFs) analyses were conducted with the aid of biological tools, MAGIA and GenCOdis3, respectively. As a result, the genes comprised of CSF1R, TYROBP, PLEK, FGR, ACLY, ACACA, LAPTM5, C1 or f162, IL10RA and CD163 were identified as hub genes. Additionally, miRNA analysis resulted in finding an association of zinc finger protein with OVCA comes out after implementing different algorithms. On the other hand, in the TFs analysis resulted in various DEGs that were enriched by NFAT, NF1 and GABP TFs. In this study, it was observed that ACACA, ACLY and CSF1R DEGs showed significant occurrence in different steps, and therefore, these genes were studied, precisely. Nevertheless, the results may help to discover the potential biomarkers with deep understanding of molecular mechanisms. However, further validation is required to explain the OVCA pathogenesis.


Cryptotanshinone inhibits oral squamous cell carcinoma through the autophagic pathway.
Oral squamous cell carcinoma (OSCC) is one of the most common malignant tumors with a low quality of life. Because traditional surgical treatment often causes large wounds and then affects the quality of life of patients, it is urgent to find new and efficient drugs with good safety for clinical treatment. This study aimed to identify potential anticancer drugs starting from the traditional Chinese medicine Salvia miltiorrhiza extract. Cryptotanshinone, a compound isolated from the Chinese herb Salvia miltiorrhiza, was found to significantly induce apoptosis and inhibit proliferation in OSCC. By electron microscopy, autophagosomes were found. Confocal fluorescence microscopy data showed that cryptotanshinone significantly induced autophagy in OSCC cells. Mechanistically, the western blot assay indicated that cryptotanshinone induced cell autophagy through the activation of the LC3 pathway, whereas the autophagy inhibitor 3-methyladenine attenuated these effects. Furthermore, we demonstrated that cryptotanshinone had a significant antitumor effect in a tumor xenograft model, and no damage to vital organs was observed. Our findings provide evidence that cryptotanshinone may be a novel therapeutic strategy for the treatment of OSCC.


Delivery of Oligonucleotide Therapeutics: Chemical Modifications, Lipid Nanoparticles, and Extracellular Vesicles.
Oligonucleotides (ONs) comprise a rapidly growing class of therapeutics. In recent years, the list of FDA-approved ON therapies has rapidly expanded. ONs are small (15-30 bp) nucleotide-based therapeutics which are capable of targeting DNA and RNA as well as other biomolecules. ONs can be subdivided into several classes based on their chemical modifications and on the mechanisms of their target interactions. Historically, the largest hindrance to the widespread usage of ON therapeutics has been their inability to effectively internalize into cells and escape from endosomes to reach their molecular targets in the cytosol or nucleus. While cell uptake has been improved, "endosomal escape" remains a significant problem. There are a range of approaches to overcome this, and in this review, we focus on three: altering the chemical structure of the ONs, formulating synthetic, lipid-based nanoparticles to encapsulate the ONs, or biologically loading the ONs into extracellular vesicles. This review provides a background to the design and mode of action of existing FDA-approved ONs. It presents the most common ON classifications and chemical modifications from a fundamental scientific perspective and provides a roadmap of the cellular uptake pathways by which ONs are trafficked. Finally, this review delves into each of the above-mentioned approaches to ON delivery, highlighting the scientific principles behind each and covering recent advances.


Study on nanoscale abrasive interaction between nanoprobe and self-assembled molecular surface for probe-based nanolithography process.
Mechano-chemical scanning probe lithography (MC-SPL) is a nanofabrication technique that is based on the nanoscale abrasive interaction between a sharp nanoprobe and an ultrathin organic resist-coated surface. This work reviews the fundamental aspects of the MC-SPL process. The abrasive interaction at the tip-workpiece contact interface that occurs in the MC-SPL process was investigated by experimentally as well as by molecular dynamics simulation. Experimental results showed that severe wear of the tip that can generate just after coming into contact with the sample and the defects on the organic resists lead to the pattern broadening. In addition, the results from the simulation using the tips modeled with various shapes disclosed that the scribed pattern width was largely affected more by the shape of the tip rather than by the tip-surface contact area and thus the severe pattern broadening and large displacement of alkanethiol molecules around the tip could occur in the case of using the tip with large cone angle. It was found that these phenomena were due to the strong van der Waals interactions between the alkanethiol molecules.


Formulation of an oral dosage form utilizing the properties of cubic liquid crystalline phases of glyceryl monooleate.
Glyceryl monooleate is a Food and Drug Administration-approved food additive which has the ability to form various liquid crystalline phases in the presence of various amounts of water. The unique properties of the cubic liquid crystalline phase that result upon the presence of excess body fluids at body temperature were utilized to formulate an oral dosage form containing furosemide as the model drug. The aim was to develop a formula, which has both bioadhesive and sustained release properties of the resultant cubic phase, so that increasing gastric residence time to improve bioavailability of the drug and at the same time obtaining a sustained action. The system was found to be affected by the limited solubility of furosemide in both the carrier system and the pH of surrounding medium. As a consequence, the addition of some solubility modifiers was investigated in order to obtain the desired properties of the expected liquid crystalline system.


V2 of tomato yellow leaf curl virus can suppress methylation-mediated transcriptional gene silencing in plants.
Tomato yellow leaf curl virus (TYLCV) is a DNA virus belonging to the genus Begomovirus. TYLCV replicates using double-stranded DNA intermediates that can become the target of plant transcriptional gene silencing (TGS). Here, we show that the V2 protein of TYLCV can suppress TGS of a transcriptionally silenced green fluorescent protein (GFP) transgene in Nicotiana benthamiana line 16-TGS. Through bisulfite sequencing and chop-PCR, we demonstrated that the TYLCV V2 can reverse GFP transgene silencing by reducing the methylation levels in the 35S promoter sequence. Both AtSN1 and MEA-ISR loci in Arabidopsis thaliana were previously reported to be strongly methylated, and we show that the methylation status of both loci was significantly reduced in TYLCV V2 transgenic Arabidopsis plants. We conclude that TYLCV can efficiently suppress TGS when it infects plants, and its V2 protein is responsible for the TGS suppression activity.


A positive consequence of the COVID-19 pandemic: how the counterfactual experience of school closures is accelerating a multisectoral response to the treatment of neglected tropical diseases.
Global access to deworming treatment is one of the public health success stories of low-income countries in the twenty-first century. Parasitic worm infections are among the most ubiquitous chronic infections of humans, and early success with mass treatment programmes for these infections was the key catalyst for the neglected tropical disease (NTD) agenda. Since the launch of the 'London Declaration' in 2012, school-based deworming programmes have become the world's largest public health interventions. WHO estimates that by 2020, some 3.3 billion school-based drug treatments had been delivered. The success of this approach was brought to a dramatic halt in April 2020 when schools were closed worldwide in response to the COVID-19 pandemic. These closures immediately excluded 1.5 billion children not only from access to education but also from all school-based health services, including deworming. WHO Pulse surveys in 2021 identified NTD treatment as among the most negatively affected health interventions worldwide, second only to mental health interventions. In reaction, governments created a global Coalition with the twin aims of reopening schools and of rebuilding more resilient school-based health systems. Today, some 86 countries, comprising more than half the world's population, are delivering on this response, and school-based coverage of some key school-based programmes exceeds those from January 2020. This paper explores how science, and a combination of new policy and epidemiological perspectives that began in the 1980s, led to the exceptional growth in school-based NTD programmes after 2012, and are again driving new momentum in response to the COVID-19 pandemic. This article is part of the theme issue 'Challenges and opportunities in the fight against neglected tropical diseases: a decade from the London Declaration on NTDs'.


A genomic region harboring the Pl1 allele from the Peruvian cultivar JC072A confers purple cob on Japanese flint corn (Zea mays L.).
Purple corn is a maize variety (Zea mays L.) with high anthocyanin content. When purple corn is used as forage, its anthocyanins may mitigate oxidative stresses causing lower milk production in dairy cows. In this study, we analyzed quantitative trait loci (QTLs) for anthocyanin pigmentation of maize organs in an F2 population derived from a cross between the Peruvian cultivar 'JC072A' (purple) and the inbred line 'Ki68' (yellowish) belonged to Japanese flint. We detected 17 significant QTLs on chromosomes 1-3, 6, and 10. Because the cob accounts for most of the fresh weight of the plant ear, we focused on a significant QTL for purple cob on chromosome 6. This QTL also conferred pigmentation of anther, spikelet, leaf sheath, culm, and bract leaf, and was confirmed by using two F3 populations. The gene Pl1 (purple plant 1) is the most likely candidate gene in this QTL region because the amino acid sequence encoded by Pl1-JC072A is similar to that of an Andean allele, Pl-bol3, which is responsible for anthocyanin production. The markers designed for the Pl1 alleles will be useful for the breeding of F1 lines with anthocyanin pigmentation in cobs.


Chip-Scale Droop-Free Fin Light-Emitting Diodes Using Facet-Selective Contacts.
Sub-micron-size light sources are currently extremely dim, achieving nanowatt output powers due to the current density and temperature droop. Recently, we reported a droop-free fin light-emitting diode (LED) pixel that at high current densities becomes a laser with record output power in the microwatt range. Here, we show a scalable method for selectively metallizing fins via their nonpolar side facet that allows electrical injection to sub-200 nm wide n-ZnO fins on p-GaN with at least 0.8 μm2 active area. Electrically addressable fin LEDs are fabricated in a linear array format using standard 2 μm resolution photolithography. Electroluminescence analysis across different pixels shows that the fin acts as the active region of the LED and generates a narrow-band ultraviolet emission between ≈368 and ≈390 nm. Investigating fins at high current densities, ranging from 100 to 2000 kA/cm2, shows that their emission increases without any decline even as the junction temperature reaches a range of 200-340 °C. The absence of electron leakage to p-GaN at high injection levels and an undetectable electron-hole escape from the fin at high temperatures indicate that the fin shape is highly efficient in controlling the nonradiative recombination pathways such as Auger recombination. The fin LED geometry is expected to enable the realization of high-brightness arrays of light sources at sub-micron-size regimes suitable for operation at high temperatures and high current densities.


The Role of Seagrass Traits in Mediating Zostera noltei Vulnerability to Mesograzers.
Understanding how intra-specific differences in plant traits mediate vulnerability to herbivores of relevant habitat-forming plants is vital to attain a better knowledge on the drivers of the structure and functioning of ecosystems. Such studies, however, are rare in seagrass-mesograzer systems despite the increasingly recognized relevance of mesograzers as seagrass consumers. We investigated the role and potential trade-offs of multiple leaf traits in mediating the vulnerability of the seagrass Zostera noltei to different mesograzer species, the amphipod Gammarus insensibilis and the isopod Idotea chelipes. We worked with plants from two different meadows for which contrasting chemical and structural traits were expected based on previous information. We found that plants with high vulnerability to mesograzers (i.e. those preferred and subjected to higher rates of leaf area loss) had not only higher nitrogen content and lower C:N, fibre, and phenolics, but also tender and thinner leaves. No trade-offs between chemical and structural traits of the seagrass were detected, as they were positively correlated. When leaf physical structure was removed using agar-reconstituted food, amphipod preference towards high-susceptibility plants disappeared; thus indicating that structural rather than chemical traits mediated the feeding preference. Removal of plant structure reduced the size of isopod preference to less than half, indicating a stronger contribution of structural traits (> 50%) but combined with chemical/nutritional traits in mediating the preference. We then hypothesized that the high environmental nutrient levels recorded in the meadow exhibiting high susceptibility modulate the differences observed between meadows in seagrass traits. To test this hypothesis, we exposed low-vulnerability shoots to eutrophic nutrient levels in a 6-week enrichment experiment. Nutrient enrichment increased Z. noltei nitrogen content and lowered C:N, fibre, and phenolics, but had no effect on structural traits. Overall, our findings help to better understand the trait-mediated seagrass susceptibility to mesograzers and reinforce the increasingly recognized role of structural defences against herbivory.


Forming a multinucleated cell: molecules that regulate myoblast fusion.
In mammals, cell fusion occurs among a limited number of cell types: sperm and oocytes during fertilization, trophoblasts during placenta formation, macrophages during giant cell and osteoclast formation and myoblasts in the formation of myofibers and myotubes. The molecular mechanisms involved in these membrane fusion events largely are unknown. This review will focus on the known molecules that regulate myoblast fusion with an emphasis on a novel signaling pathway involving the calcium-regulated transcription factor NFATC2 in the regulation of myoblast fusion.


Study of 5-HT release with a chronically implanted microdialysis probe in the ventral horn of the spinal cord of unrestrained rats during exercise on a treadmill.
The ventral horn of the spinal cord is profusely innervated by serotonin (5-hydroxytryptamine, 5-HT) which presumably modulates locomotor activity through motoneurons. However, direct evidence of correlation between 5-HT release and activation of motoneurons is still lacking. In order to appreciate the functional characteristics of this innervation, we have used microdialysis to monitor the release of 5-HT in the spinal cord of rats spontaneously running on a treadmill. For this purpose, we developed an original surgical procedure adapted for the chronic implantation of a microdialysis probe in the lumbar spinal cord. The probe was kept in place for 40 days, and microdialysis experiments were carried out at days 8, 16, 20 and 32. 5-HT was detected with HPLC coupled to electrochemistry. This technique demonstrated that release of 5-HT is not increased during exercise. However, a significant decrease was measured during postexercise rest. 5-HT could still be detected 32 days after probe implantation. Detailed histological and immunocytochemical analysis based on glial fibrillary acidic protein and 5-HT immunocytochemistry showed minimal gliosis and the presence of serotonergic varicose fibers, respectively, at the probe contact. It thus appears that microdialysis can be performed through a probe implanted chronically in the spinal cord of unrestrained rat during an endurance running exercise.


Non-preferred contrast responses in the Drosophila motion pathways reveal a receptive field structure that explains a common visual illusion.
Diverse sensory systems, from audition to thermosensation, feature a separation of inputs into ON (increments) and OFF (decrements) signals. In the Drosophila visual system, separate ON and OFF pathways compute the direction of motion, yet anatomical and functional studies have identified some crosstalk between these channels. We used this well-studied circuit to ask whether the motion computation depends on ON-OFF pathway crosstalk. Using whole-cell electrophysiology, we recorded visual responses of T4 (ON) and T5 (OFF) cells, mapped their composite ON-OFF receptive fields, and found that they share a similar spatiotemporal structure. We fit a biophysical model to these receptive fields that accurately predicts directionally selective T4 and T5 responses to both ON and OFF moving stimuli. This model also provides a detailed mechanistic explanation for the directional preference inversion in response to the prominent reverse-phi illusion. Finally, we used the steering responses of tethered flying flies to validate the model's predicted effects of varying stimulus parameters on the behavioral turning inversion.


Risk and Respirators: The Hazardous Trajectories of Soviet Occupational Safety, 1940s-80s.
This article explores a distinct Soviet policy of occupational care that emerged after World War II, when Soviet industry invented an array of respiratory protective equipment (RPE). The annual production of millions of devices highlighted the development of a complex Soviet transinstitutional system for insuring safe occupational breathing. Following key premises of respiratory safety policies in the activities of Soviet organizations, this article traces a biopolitical shift from mortality to vitality. By showing the curial role of RPE in the history of occupational rather than military safety in the twentieth century, the article fills a major research gap to feature Soviet modernity through the unique lens of industrial respiratory care in postwar Soviet Union between the late 1940s and the early 1990s.


A Drug Repurposing Approach for Antimalarials Interfering with SARS-CoV-2 Spike Protein Receptor Binding Domain (RBD) and Human Angiotensin-Converting Enzyme 2 (ACE2).
Host cell invasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is mediated by the interaction of the viral spike protein (S) with human angiotensin-converting enzyme 2 (ACE2) through the receptor-binding domain (RBD). In this work, computational and experimental techniques were combined to screen antimalarial compounds from different chemical classes, with the aim of identifying small molecules interfering with the RBD-ACE2 interaction and, consequently, with cell invasion. Docking studies showed that the compounds interfere with the same region of the RBD, but different interaction patterns were noted for ACE2. Virtual screening indicated pyronaridine as the most promising RBD and ACE2 ligand, and molecular dynamics simulations confirmed the stability of the predicted complex with the RBD. Bio-layer interferometry showed that artemisone and methylene blue have a strong binding affinity for RBD (KD = 0.363 and 0.226 μM). Pyronaridine also binds RBD and ACE2 in vitro (KD = 56.8 and 51.3 μM). Overall, these three compounds inhibit the binding of RBD to ACE2 in the μM range, supporting the in silico data.


Energy requirements for maintenance of crossbred beef cattle with different genetic potential for milk.
Maintenance energy requirements were estimated in two gestation and one lactation feeding trials for three groups of cows. All cows in the second gestation trial completed the first gestation and lactation trials. The three groups were chosen to represent cattle similar in growth rate and mature size but different in amount of milk provided to their calves. The low (L) group included Hereford x Angus, the medium (M) group included Red Poll x Angus and the high (H) group included Milking Shorthorn x Angus cows. Cows were individually fed to maintain net body weight (minus gravid uterus for gestation) constant. Allowances were made in energy intake for gestation and lactation. Cow weights were adjusted to an average condition score in each trial. Daily maintenance requirements during gestation were 18% lower than those during lactation. The H and M cows required 12% more energy per unit metabolic weight than L cows to maintain body weight during both gestation and lactation. Differences in milk production explained 23% of the variation in maintenance requirements, suggesting that important differences exist beyond those associated with milk production potential. Repeatabilities of maintenance requirement measurements ranged from .44 to .64. Maintenance requirements for calves under feedlot conditions in the postweaning phase were estimated from data collected from 494 calves, half-sibs and offspring of the cows described previously. Energy requirements were 11% higher for the H and M groups than for the L group.


Jeavons syndrome existing as occipital cortex initiating generalized epilepsy.
Jeavons syndrome (JS) is one of the underreported epileptic syndromes characterized by eyelid myoclonia (EM), eye closure-induced seizures/electroencephalography (EEG) paroxysms, and photosensitivity. JS has been proposed as idiopathic generalized epilepsy (IGE) because of normal posterior dominant background activity and paroxysmal generalized ictal epileptiform discharges (EDs). However, we noticed subtle occipital EDs preceding EM and interictal posterior EDs using digital video-EEG. We studied clinical and EEG findings in JS to determine the specific occipital lobe relation to this "eye closure-induced" reflex IGE. We identified 12 children who met the diagnostic criteria of JS from January 2004 to April 2009 at the Hospital for Sick Children, Toronto, Canada. All patients had EM captured by video-EEG. We reviewed and described ictal EEG patterns, interictal abnormalities, and demographics, clinical, and neuroimaging findings. All patients but one were female (92%). Age at seizure onset ranged from 1.5 to 9 years, with a mean age of 4.9 years. Six patients (50%) were previously diagnosed as having absence epilepsy and 10 patients were on antiepileptic medications. All 12 patients had normal posterior dominant alpha rhythm, reactive to eye opening and closure. Spiky posterior alpha activity was noted with sustained eye closure in six patients (50%). Interictally, there were generalized EDs found in 10 patients (83%); four of them also had focal interictal EDs over the posterior head region. Eleven patients (92%) had evidence of focal posterior ictal EDs. EM and/or paroxysmal EDs were induced by photic stimulation in 9 (75%) and hyperventilation in 7 (58%). We observed two neurophysiologic findings in JS: (1) focal interictal EDs from posterior head region; and (2) predominant focal posterior ictal EDs preceding generalized EDs. Further clinical observations of seizures induced by eye closure, photic stimulation, and hyperventilation along with EEG paroxysms would raise the possibility of the occipital cortex initiating generalized epilepsy network involving the brainstem, and thalamocortical and transcortical pathways in JS.


The E3 ubiquitin ligase NEDD4 is an LC3-interactive protein and regulates autophagy.
The MAP1LC3/LC3 family plays an essential role in autophagosomal biogenesis and transport. In this report, we show that the HECT family E3 ubiquitin ligase NEDD4 interacts with LC3 and is involved in autophagosomal biogenesis. NEDD4 binds to LC3 through a conserved WXXL LC3-binding motif in a region between the C2 and the WW2 domains. Knockdown of NEDD4 impaired starvation- or rapamycin-induced activation of autophagy and autophagosomal biogenesis and caused aggregates of the LC3 puncta colocalized with endoplasmic reticulum membrane markers. Electron microscopy observed gigantic deformed mitochondria in NEDD4 knockdown cells, suggesting that NEDD4 might function in mitophagy. Furthermore, SQSTM1 is ubiquitinated by NEDD4 while LC3 functions as an activator of NEDD4 ligase activity. Taken together, our studies define an important role of NEDD4 in regulation of autophagy.


Should tourniquets be used in upper limb surgery? A systematic review and meta-analysis.
This study compares the intra- and post-operative outcomes of upper limb orthopaedic surgical procedures performed with and without tourniquet assistance. A systematic review was undertaken assessing the electronic databases Medline, CINAHL, AMED and EMBASE. The evidence-base was critically appraised using the Cochrane Bone, Joint and Muscle Trauma Group quality assessment tool. Study heterogeneity was statistically tested using Chi2 and I2 statistics. Where appropriate a random-effects meta-analysis was undertaken to pool results of primary studies assessing mean difference of each outcome. Two studies investigating fifty-five patients undergoing upper limb surgery were identified. The limited findings suggest that the use of tourniquets may reduce the incidence of technical difficulties during upper limb surgery. It remains unclear whether the application of a tourniquet can influence pain perception or operative duration. The evidence-base was considerably limited in both size and methodological quality. Further study is recommended to address the literature's methodological weaknesses.


Halogen Bonding in Sulphonamide Co-Crystals: X···π Preferred over X···O/N?
Sulphonamides have been one of the major pharmaceutical compound classes since their introduction in the 1930s. Co-crystallisation of sulphonamides with halogen bonding (XB) might lead to a new class of pharmaceutical-relevant co-crystals. We present the synthesis and structural analysis of seven new co-crystals of simple sulphonamides N-methylbenzenesulphonamide (NMBSA), N-phenylmethanesulphonamide (NPMSA), and N-phenylbenzenesulphonamide (BSA), as well as of an anti-diabetic agent Chlorpropamide (CPA), with the model XB-donors 1,4-diiodotetrafluorobenzene (14DITFB), 1,4-dibromotetrafluorobenzene (14DBTFB), and 1,2-diiodotetrafluorobenzene (12DITFB). In the reported co-crystals, X···O/N bonds do not represent the most common intermolecular interaction. Against our rational design expectations and the results of our statistical CSD analysis, the normally less often present X···π interaction dominates the crystal packing. Furthermore, the general interaction pattern in model sulphonamides and the CPA multicomponent crystals differ, mainly due to strong hydrogen bonds blocking possible interaction sites.


Do Sensory Stimulation Programs Have an Impact on Consciousness Recovery?
Objectives: Considering sensory stimulation programs (SSP) as a treatment for disorders of consciousness is still debated today. Previous studies investigating its efficacy were affected by various biases among which small sample size and spontaneous recovery. In this study, treatment-related changes were assessed using time-series design in patients with disorders of consciousness (i.e., vegetative state-VS and minimally conscious state-MCS). Methods: A withdrawal design (ABAB) was used. During B phases, patients underwent a SSP (3 days a week, including auditory, visual, tactile, olfactory, and gustatory stimulation). The program was not applied during A phases. To assess behavioral changes, the Coma Recovery Scale-Revised (CRS-R) was administered by an independent rater on a weekly basis, across all phases. Each phase lasted 4 weeks. In a subset of patients, resting state functional magnetic resonance imaging (fMRI) data were collected at the end of each phase. Results: Twenty nine patients (48 ± 19 years old; 15 traumatic; 21 > a year post-injury; 11 VS and 18 MCS) were included in our study. Higher CRS-R total scores (medium effect size) as well as higher arousal and oromotor subscores were observed in the B phases (treatment) as compared to A phases (no treatment), in the MCS group but not in the VS group. In the three patients who underwent fMRI analyses, a modulation of metabolic activity related to treatment was observed in middle frontal gyrus, superior temporal gyrus as well as ventro-anterior thalamic nucleus. Conclusion: Our results suggest that SSP may not be sufficient to restore consciousness. SSP might nevertheless lead to improved behavioral responsiveness in MCS patients. Our results show higher CRS-R total scores when treatment is applied, and more exactly, increased arousal and oromotor functions.


Substance use progression from adolescence to early adulthood: effortful control in the context of friendship influence and early-onset use.
In a sample of 998 ethnically diverse adolescents, a multiagent, multimethod approach to the measurement of adolescent effortful control, adolescent substance use, and friendship influence was used to predict escalations to early-adult tobacco, alcohol, and marijuana use by ages 22-23. Structural equation modeling revealed that adolescent substance use and friends' substance use tended to be highly correlated and together were robust predictors of a problematic pattern of usage for all substances in early adulthood. In addition, the adolescent effortful control construct directly predicted progressions to problematic use of tobacco and marijuana, but not for alcohol. In the alcohol model, effortful control interacted with the construct of substance use lifestyle (based on adolescent alcohol use and friends' substance use) when predicting problematic alcohol use in early adulthood. Results held when comparing across genders and across ethnic groups. These findings emphasize the importance of addressing adolescent self-regulation in interventions designed to treat and prevent early-adult substance abuse.


Subsensitivity of bronchodilator and systemic beta 2 adrenoceptor responses after regular twice daily treatment with eformoterol dry powder in asthmatic patients.
There is controversy as to the role of long acting beta 2 agonists such as eformoterol and, in particular, whether bronchodilator tolerance occurs during continuous therapy. The purpose of this study was to extend previous observations of bronchodilator subsensitivity with metered dose eformoterol aerosol in order to assess whether tolerance also occurs with a dry powder formulation of the same drug. Sixteen asthmatic patients of mean age 33 (range 18-53) years and FEV1 (% predicted) of 64 (3)%, of whom 13 were receiving inhaled corticosteroids, received regular treatment with eformoterol 24 micrograms twice daily or placebo twice daily (without beta 2 agonists) given concurrently for four weeks in a randomised double blind cross-over design. An initial two week run-in was used when beta 2 agonists were withdrawn and substituted with ipratropium bromide. Dose-response curves to eformoterol (cumulative dose 6-102 micrograms) for airways and systemic beta 2 responses were constructed at the end of each treatment period. Baseline values for airways and systemic responses were similar. The peak delta FEV1 response from the dose-response curve (as change from baseline) and the delta response for FEV1 and FEF25-75 at six hours after the last dose were attenuated after eformoterol compared with placebo: peak delta FEV1 response 1.001 with placebo v 0.841 with eformoterol (95% CI 0.04 to 0.28); at six hours 0.931 with placebo v 0.581 with eformoterol (95% CI 0.20 to 0.50); and for delta FEF25-75 at six hours 1.29 1/s with placebo v 0.87 1/s with eformoterol (95% CI 0.15 to 0.69). Morning peak expiratory flow rate was significantly improved during treatment with eformoterol (451 1/min) compared with placebo (399 1/min) (95% CI 21 to 82). Systemic beta 2 responses were blunted after eformoterol, together with a reduction in lymphocyte beta 2 receptor binding density. Regular twice daily eformoterol dry powder may produce bronchodilator subsensitivity in terms of both peak and duration of response to cumulative repeated doses of eformoterol. Systemic beta 2-mediated adverse effects also showed tolerance, which was mirrored by downregulation of lymphocyte beta 2 adrenoceptors.


Factors associated with pain and osteoarthritis at the hip and knee in Great Britain's Olympians: a cross-sectional study.
Knowledge of the epidemiology and potentially modifiable factors associated with musculoskeletal disease is an important first step in injury prevention among elite athletes. This study investigated the prevalence and factors associated with pain and osteoarthritis (OA) at the hip and knee in Great Britain's (GB) Olympians aged 40 and older. This is a cross-sectional study. A survey was distributed to 2742 GB Olympians living in 30 countries. Of the 714 (26.0%) who responded, 605 were eligible for analysis (ie, aged 40 and older). The prevalence of hip and knee pain was 22.4% and 26.1%, and of hip and knee OA was 11.1% and 14.2%, respectively. Using a multivariable model, injury was associated with OA at the hip (adjusted OR (aOR) 10.85; 95% CI 3.80 to 30.96) and knee (aOR 4.92; 95% CI 2.58 to 9.38), and pain at the hip (aOR 5.55; 95% CI 1.83 to 16.86) and knee (aOR 2.65; 95% CI 1.57 to 4.46). Widespread pain was associated with pain at the hip (aOR 7.63; 95% CI 1.84 to 31.72) and knee (aOR 4.77; 95% CI 1.58 to 14.41). Older age, obesity, knee malalignment, comorbidities, hypermobility and weight-bearing exercise were associated with hip and knee OA and/or pain. This study detected an association between several factors and hip and knee pain/OA in retired GB Olympic athletes. These associations require further substantiation in retired athletes from other National Olympic Committees, and through comparison with the general population. Longitudinal follow-up is needed to investigate the factors associated with the onset and progression of OA/pain, and to determine if modulation of such factors can reduce the prevalence of pain and OA in this population.


Evaluation of the Abbott RealTime MTB and RealTime MTB INH/RIF Assays for Direct Detection of Mycobacterium tuberculosis Complex and Resistance Markers in Respiratory and Extrapulmonary Specimens.
The Abbott RealTime MTB (RT MTB) assay is a new automated nucleic acid amplification test for the detection of Mycobacterium tuberculosis complex (MTBC) in clinical specimens. In combination with the RealTime MTB INH/RIF (RT MTB INH/RIF) resistance assay, which can be applied to RT MTB-positive specimens as an add-on assay, the tests also indicate the genetic markers of resistance to isoniazid (INH) and rifampin (RIF). We aimed to evaluate the diagnostic sensitivity and specificity of RT MTB using different types of respiratory and extrapulmonary specimens and to compare performance characteristics directly with those of the FluoroType MTB assay. The resistance results obtained by RT MTB INH/RIF were compared to those from the GenoType MTBDRplus and from phenotypic drug susceptibility testing. A total of 715 clinical specimens were analyzed. Compared to culture, the overall sensitivity of RT MTB was 92.1%; the sensitivity rates for smear-positive and smear-negative samples were 100% and 76.2%, respectively. The sensitivities of smear-negative specimens were almost identical for respiratory (76.3%) and extrapulmonary (76%) specimens. Specificity rates were 100% and 95.8% for culture-negative specimens and those that grew nontuberculous mycobacteria, respectively. RT MTB INH/RIF was applied to 233 RT MTB-positive samples and identified resistance markers in 7.7% of samples. Agreement with phenotypic and genotypic drug susceptibility testing was 99.5%. In conclusion, RT MTB and RT MTB INH/RIF allow for the rapid and accurate diagnosis of tuberculosis (TB) in different types of specimens and reliably indicate resistance markers. The strengths of this system are the comparably high sensitivity with paucibacillary specimens, its ability to detect INH and RIF resistance, and its high-throughput capacities.


Differential influence of vitamin C on the peripheral and cerebral circulation after diving and exposure to hyperoxia.
We examined if the diving-induced vascular changes in the peripheral and cerebral circulation could be prevented by oral antioxidant supplementation. Fourteen divers performed a single scuba dive to eighteen meter sea water for 47 min. Twelve of the divers participated in a follow-up study involving breathing 60% of oxygen at ambient pressure for 47 min. Before both studies, participants ingested vitamin C (2 g/day) or a placebo capsule for 6 days. After a 2-wk washout, the study was repeated with the different condition. Endothelium-dependent vasodilator function of the brachial artery was assessed pre- and postintervention using the flow-mediated dilation (FMD) technique. Transcranial Doppler ultrasound was used to measure intracranial blood velocities pre- and 90 min postintervention. FMD was reduced by ∼32.8% and ∼21.2% postdive in the placebo and vitamin C trial and posthyperoxic condition in the placebo trial by ∼28.2% ( P < 0.05). This reduction in FMD was attenuated by ∼10% following vitamin C supplementation in the hyperoxic study ( P > 0.05). Elevations in intracranial blood velocities 30 min after surfacing from diving were reduced in the vitamin C study compared with the placebo trial ( P < 0.05). O2 breathing had no postintervention effects on intracranial velocities ( P > 0.05). Prophylactic ingestion of vitamin C effectively abrogated peripheral vascular dysfunction following exposure to 60% O2 but did not abolish the postdive decrease in FMD. Transient elevations of intracranial velocities postdive were reduced by vitamin C. These findings highlight the differential influence of vitamin C on peripheral and cerebral circulations following scuba diving, which are only partly mediated via hyperoxia.


New indices in scholarship assessment.
Faculty members' contributions to research and scholarship are measured by a variety of indices. Assessment also has become an integral part of the Accreditation Council for Pharmacy Education's accreditation process for professional programs. This review describes some of the newer indices available for faculty scholarship assessment. Recently described metrics include the h-index, m-quotient, g-index, h(2) index, a-index, m-index, r-index, ar index, and the creativity index. Of the newer scholarship metrics available, the h-index and m-quotient will likely have the most widespread application in the near future. However, there is no substitute for thoughtful peer review by experienced academicians as the primary method of research and scholarship assessment.


Genome location and identification of functions defective in the Bartha vaccine strain of pseudorabies virus.
We have shown previously (Lomniczi et al., J. Virol. 52:198-205, 1984) that the Bartha vaccine strain of pseudorabies virus has a deletion in the short unique (Us) region of its genome--a deletion that is related to the absence of virus virulence. This strain is, however, also defective in other genes involved in virulence. We show here that virulence can be restored by marker rescue of the Bartha strain to which an intact Us has been restored (but not to the parental Bartha strain) by sequences derived from approximate map units 0.460 and 0.505 of the wild-type virus genome. No difference in the ability to grow in cell culture was observed between parental Bartha, Bartha 43/25a (Bartha to which an intact Us has been restored), or the doubly rescued Bartha strains. However, only the doubly rescued Bartha strain was virulent for both chickens and pigs and replicated to high titers when inoculated directly into the brains of chickens. The sequences that could restore virulence to the Bartha 43/25a strain encode four genes, all of which are involved in processes leading to the assembly of nucleocapsids. Since these sequences rescue virulence, it appears that a function that plays a role in nucleocapsid assembly is defective in the Bartha strain and that this defect contributes to the lack of virulence of this virus.


Effect of bottlenecking on evolution of the nonstructural protein 3 gene of hepatitis C virus during sexually transmitted acute resolving infection.
Sexual partners of patients infected with the hepatitis C virus (HCV) often have detectable HCV-specific T-cell responses in the absence of seroconversion, suggesting unapparent, spontaneously resolving infection. To determine whether differences in the evolutionary potential of bottlenecked inoculum may explain the low rate of HCV persistence after sexual exposure, we have investigated changes in the entire HCV nonstructural 3 (NS3) gene over time in a chronic carrier and compared his viral quasispecies with that of the acute-phase isolate of his sexual partner, who developed acute resolving hepatitis C. The overall rate of accumulation of mutations, estimated by regression analysis of six consecutive consensus NS3 sequences over 8 years, was 1.5 x 10(-3) mutations per site per year, with small intersample fluctuations related to changes in environmental conditions. Comparison of quasispecies parameters in one isolate of the chronic carrier with those of the acute-phase isolate of the infected partner revealed a higher heterogeneity and lower proportion of nonsynonymous mutations in the former. All NS3 sequences from the acute-phase isolate clustered with a single sequence from the chronic isolate, despite complete HLA mismatch between the patients, suggesting bottlenecking during transmission. The low risk of viral persistence after sexual exposure to HCV may be related to the selection of a limited number of viral particles carrying a particular combination of mutations which may further limit the potential of a relatively homogeneous quasispecies to rapidly diversify and overcome the immune response of the exposed host.


ECONOMICS NOBEL: Dealing With Biases and Discrete Choices.
This year's Bank of Sweden Prize in Economic Sciences, given in honor of Alfred Nobel, goes to two researchers who gave the field of microeconomics--the study of individuals' economic behavior--new tools to help draw conclusions from imperfect data. James Heckman of the University of Chicago wins half of this year's prize for coming up with ways to deal with selection biases. Daniel McFadden of the University of California, Berkeley, tackled a different conundrum: how to quantify discrete choices rather than continuous ones.


Responses to Bacteria, Virus, and Malaria Distinguish the Etiology of Pediatric Clinical Pneumonia.
Plasma-detectable biomarkers that rapidly and accurately diagnose bacterial infections in children with suspected pneumonia could reduce the morbidity of respiratory disease and decrease the unnecessary use of antibiotic therapy. Using 56 markers measured in a multiplexed immunoassay, we sought to identify proteins and protein combinations that could discriminate bacterial from viral or malarial diagnoses. We selected 80 patients with clinically diagnosed pneumonia (as defined by the World Health Organization) who also met criteria for bacterial, viral, or malarial infection based on clinical, radiographic, and laboratory results. Ten healthy community control subjects were enrolled to assess marker reliability. Patients were subdivided into two sets: one for identifying potential markers and another for validating them. Three proteins (haptoglobin, tumor necrosis factor receptor 2 or IL-10, and tissue inhibitor of metalloproteinases 1) were identified that, when combined through a classification tree signature, accurately classified patients into bacterial, malarial, and viral etiologies and misclassified only one patient with bacterial pneumonia from the validation set. The overall sensitivity and specificity of this signature for the bacterial diagnosis were 96 and 86%, respectively. Alternative combinations of markers with comparable accuracy were selected by support vector machine and regression models and included haptoglobin, IL-10, and creatine kinase-MB. Combinations of plasma proteins accurately identified children with a respiratory syndrome who were likely to have bacterial infections and who would benefit from antibiotic therapy. When used in conjunction with malaria diagnostic tests, they may improve diagnostic specificity and simplify treatment decisions for clinicians.


Pancreatic acini possess endothelin receptors whose internalization is regulated by PLC-activating agents.
Endothelin-1 (ET-1) and ET-3 mRNA have been found in the pancreas. We investigated the ability of ET-1, ET-2, and ET-3 to interact with and alter dispersed rat pancreatic acinar cell function. Radiolabeled ETs bound in a time- and temperature-dependent fashion, which was specific and saturable. Analysis demonstrated two classes of receptors, one class (ETA receptor) had a high affinity for ET-1 but a low affinity for ET-3, and the other class (ETB receptor) had equally high affinities for ET-1 and ET-3. No specific receptor for ET-2 was identified. Pancreatic secretagogues that activate phospholipase C (PLC) inhibited binding of 125I-labeled ET-1 (125I-ET-1) or 125I-ET-3, whereas agents that act through adenosine 3',5'-cyclic monophosphate (cAMP) did not. A23187 had no effect on 125I-ET-1 or 125I-ET-3 binding, whereas the phorbol ester 12-O-tetradecanoylphorbol 13-acetate reduced binding. The effect of cholecystokinin octapeptide (CCK-8) was mediated through its own receptor. Stripping of surface bound ligand studies demonstrated that both 125I-labeled ET-1 and 125I-labeled ET-3 were rapidly internalized. CCK-8 decreased the internalization but did not change the amount of surface bound ligand. Endothelins neither stimulate nor alter changes in enzyme secretion, intracellular calcium, cAMP, or [3H]inositol trisphosphate (IP3). This study demonstrates the presence of ETA and ETB receptors on rat pancreatic acini; occupation of both receptors resulted in rapid internalization, which is regulated by PLC-activating secretagogues. Occupation of either ET receptor did not alter intracellular calcium, cAMP, IP3, or stimulate amylase release.


How Tilting the Head Interferes With Eye-Hand Coordination: The Role of Gravity in Visuo-Proprioceptive, Cross-Modal Sensory Transformations.
To correctly position the hand with respect to the spatial location and orientation of an object to be reached/grasped, visual information about the target and proprioceptive information from the hand must be compared. Since visual and proprioceptive sensory modalities are inherently encoded in a retinal and musculo-skeletal reference frame, respectively, this comparison requires cross-modal sensory transformations. Previous studies have shown that lateral tilts of the head interfere with the visuo-proprioceptive transformations. It is unclear, however, whether this phenomenon is related to the neck flexion or to the head-gravity misalignment. To answer to this question, we performed three virtual reality experiments in which we compared a grasping-like movement with lateral neck flexions executed in an upright seated position and while lying supine. In the main experiment, the task requires cross-modal transformations, because the target information is visually acquired, and the hand is sensed through proprioception only. In the other two control experiments, the task is unimodal, because both target and hand are sensed through one, and the same, sensory channel (vision and proprioception, respectively), and, hence, cross-modal processing is unnecessary. The results show that lateral neck flexions have considerably different effects in the seated and supine posture, but only for the cross-modal task. More precisely, the subjects' response variability and the importance associated to the visual encoding of the information significantly increased when supine. We show that these findings are consistent with the idea that head-gravity misalignment interferes with the visuo-proprioceptive cross-modal processing. Indeed, the principle of statistical optimality in multisensory integration predicts the observed results if the noise associated to the visuo-proprioceptive transformations is assumed to be affected by gravitational signals, and not by neck proprioceptive signals per se. This finding is also consistent with the observation of otolithic projections in the posterior parietal cortex, which is involved in the visuo-proprioceptive processing. Altogether these findings represent a clear evidence of the theorized central role of gravity in spatial perception. More precisely, otolithic signals would contribute to reciprocally align the reference frames in which the available sensory information can be encoded.


[Field study of nursing care quality. Interaction and asepsis in clinical practice].
Competence as communication skills and as skilled practice of asepsis were studied by observing four nurses while interacting with patients and performing intravenous procedures. Nurses were observed using sterile equipment for methods of intravenous therapy. Asepsis is performed frequently, but through misunderstanding in the learning of asepsis or improper model learning the nurses may establish incorrect routines. When performing procedures, unexpected factors can distract both the expert and the inexperienced, resulting in a failure to apply basic aseptic techniques. The nurses showed an interest in the patient by listening and giving responses. Nurses may control interactions with the patient by using undesirable communication skills which include incomplete sentences, incomplete explanations and closed questions. Asked to evaluate their own behavior, the nurses did not estimate whether or not the appropriate skills were applied in observed situations. To improve the quality of nursing care performance it is recommended to further develop and apply skill training programs.


Use of in situ solid-phase adsorption in microbial natural product fermentation development.
It has been half a century since investigators first began experimenting with adding ion exchange resins during the fermentation of microbial natural products. With the development of nonionic polymeric adsorbents in the 1970s, the application of in situ product adsorption in bioprocessing has grown slowly, but steadily. To date, in situ product adsorption strategies have been used in biotransformations, plant cell culture, the production of biofuels, and selected bulk chemicals, such as butanol and lactic acid, as well as in more traditional natural product fermentation within the pharmaceutical industry. Apart from the operational gains in efficiency from the integration of fermentation and primary recovery, the addition of adsorbents during fermentation has repeatedly demonstrated the capacity to significantly increase titers by sequestering the product and preventing or mitigating degradation, feedback inhibition and/or cytotoxic effects. Adoption of in situ product adsorption has been particularly valuable in the early stages of natural product-based drug discovery programs, where quickly and cost-effectively generating multigram quantities of a lead compound can be challenging when using a wild-type strain and fermentation conditions that have not been optimized. While much of the literature involving in situ adsorption describes its application early in the drug development process, this does not imply that the potential for scale-up is limited. To date, commercial-scale processes utilizing in situ product adsorption have reached batch sizes of at least 30,000 l. Here we present examples where in situ product adsorption has been used to improve product titers or alter the ratios among biosynthetically related natural products, examine some of the relevant variables to consider, and discuss the mechanisms by which in situ adsorption may impact the biosynthesis of microbial natural products.


Dielectrophoretic placement of quasi-zero-, one-, and two-dimensional nanomaterials into nanogap for electrical characterizations.
DEP is one of promising techniques for positioning nanomaterials into the desirable location for nanoelectronic applications. In contrast, the lithography technique is commonly used to make ultra-thin conducting wires and narrow gaps but, due to the limit of patterning resolution, it is not feasible to make electrical contacts on ultra-small nanomaterials for a bottom-up device fabrication. Thus, integrating the lithography and dielectrophoresis, a real bottom-up fabrication can be achieved. In this work, the device with the nanogap in between two nanofinger-electrodes is made using electron-beam lithography from top down and the ultra-small nanomaterials, such as colloidal PbSe quantum dots, polyaniline nanofibers, and reduced-graphene-oxide flakes, are placed in the nanogap by DEP from bottom up. The threshold electric field for the DEP placement of PbSe nanocrystals was roughly estimated to be about 8.3 × 10(4) V/cm under our experimental configuration. After the DEP process, several procedures for reducing contact resistances are attempted and measurements of intrinsic electron transport in versatile nanomaterials are performed. It is experimentally confirmed that electron transport in both PbSe nanocrystal arrays and polyaniline nanofibers agrees well with Prof. Ping Sheng's model of granular metallic conduction. In addition, electron transport in reduced-graphene-oxide flakes follows Mott's 2D variable-range-hopping model. This study illustrates an integration of the electron-beam lithography and the DEP techniques for a precise manipulation of nanomaterials into electronic circuits for characterization of intrinsic properties.


The effect of A 2A receptor antagonist (SCH 442416) on the mRNA expression of Kir 2.1 and Kir 4.1 channels in rat retinal Müller cells under hypoxic conditions in vitro.
To investigate the situation of inwardly rectifying potassium channels Kir 2.1 and Kir 4.1 under hypo-xic conditions, and whether the A2A receptor antagonist could modulate the mRNA expression of Kir 2.1 and Kir 4.1 channels in retinal Müller cells in vitro under hypoxic conditions. Müller cells were treated with 0.1 1, 10 and 100 µM of A2A receptor antagonist (SCH 442416) under hypoxic conditions for 24 h, and the expression of the Kir 2.1 and Kir 4.1 mRNA channels was examined using real-time polymerase chain reaction (qPCR). There were no significant changes in the mRNA expression of the Kir 2.1 or Kir 4.1 channels under hypoxic conditions compared with normal conditions after 24 h cultured in vitro. The mRNA expression of Kir 2.1 and Kir 4.1 channels treated with 0.1 µM SCH 442416 under hypoxic conditions were increased, but at higher concentrations of SCH 442416, the mRNA expression of the Kir 2.1 and Kir 4.1 channels decreased. The A2A receptor antagonist (SCH 442416) could increase mRNA expression of the Kir 2.1and Kir 4.1 channels in Müller cells to protect the retinal neurons in vitro under hypoxic conditions.


ASCL2 reciprocally controls key trophoblast lineage decisions during hemochorial placenta development.
Invasive trophoblast cells are critical to spiral artery remodeling in hemochorial placentation. Insufficient trophoblast cell invasion and vascular remodeling can lead to pregnancy disorders including preeclampsia, preterm birth, and intrauterine growth restriction. Previous studies in mice identified achaete-scute homolog 2 (ASCL2) as essential to extraembryonic development. We hypothesized that ASCL2 is a critical and conserved regulator of invasive trophoblast cell lineage development. In contrast to the mouse, the rat possesses deep intrauterine trophoblast cell invasion and spiral artery remodeling similar to human placentation. In this study, we investigated invasive/extravillous trophoblast (EVT) cell differentiation using human trophoblast stem (TS) cells and a loss-of-function mutant Ascl2 rat model. ASCL2 transcripts are expressed in the EVT column and junctional zone, which represent tissue sources of invasive trophoblast progenitor cells within human and rat placentation sites, respectively. Differentiation of human TS cells into EVT cells resulted in significant up-regulation of ASCL2 and several other transcripts indicative of EVT cell differentiation. Disruption of ASCL2 impaired EVT cell differentiation, as indicated by cell morphology and transcript profiles. RNA sequencing analysis of ASCL2-deficient trophoblast cells identified both down-regulation of EVT cell-associated transcripts and up-regulation of syncytiotrophoblast-associated transcripts, indicative of dual activating and repressing functions. ASCL2 deficiency in the rat impacted placental morphogenesis, resulting in junctional zone dysgenesis and failed intrauterine trophoblast cell invasion. ASCL2 acts as a critical and conserved regulator of invasive trophoblast cell lineage development and a modulator of the syncytiotrophoblast lineage.