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Richard E RobertsSchool of Biomedical Sciences, University of Nottingham, Nottingham, United KingdomAbstract: Hypertension is a risk factor for myocardial infarction, stroke, renal failure, heart failure, and peripheral vascular disease. One feature of hypertension is a hyperresponsiveness to contractile agents, and inhibition of vasoconstriction forms the basis of some of the treatments for hypertension. Hypertension is also associated with an increase in the growth and proliferation of vascular smooth muscle cells, which can lead to a thickening of the smooth muscle layer of the blood vessels and a reduction in lumen diameter. Targeting both the enhanced contractile responses, and the increased vascular smooth muscle cell growth could potentially be an important pharmacological treatment of hypertension. Extracellular signal-regulated kinase (ERK) is a member of the mitogen-activated protein kinase family that is involved in both vasoconstriction and vascular smooth muscle cell growth and this, therefore, makes it an attractive therapeutic target for treatment of hypertension. ERK activity is raised in vascular smooth muscle cells from animal models of hypertension, and inhibition of ERK activation reduces both vascular smooth muscle cell growth and vasoconstriction. This review discusses the potential for targeting ERK activity in the treatment of hypertension.Keywords: ERK, hypertension, smooth muscle, vasoconstriction

Laser threshold magnetometry using the negatively charged nitrogen-vacancy (NV−) centre in diamond as a gain medium has been proposed as a technique to dramatically enhance the sensitivity of room-temperature magnetometry. We experimentally explore a diamond-loaded open tunable fibre-cavity system as a potential contender for the realisation of lasing with NV− centres. We observe amplification of the transmission of a cavity-resonant seed laser at 721 nm when the cavity is pumped at 532 nm and attribute this to stimulated emission. Changes in the intensity of spontaneously emitted photons accompany the amplification, and a qualitative model including stimulated emission and ionisation dynamics of the NV− centre captures the dynamics in the experiment very well. These results highlight important considerations in the realisation of an NV− laser in diamond.

Background and Purpose: Stimulation of vascular beta-adrenoceptors causes vasodilatation through activation of adenylyl cyclase (AC) and plasma membrane potassium channels, and beta-adrenoceptors have been linked to activation of extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase in various cell lines. However, how these findings relate to functional responses in intact tissues is largely unknown. The aim of this study, therefore, was to investigate the role of ERK in beta-adrenoceptor-induced vasodilatation.Experimental Approach: Segments of porcine coronary artery were mounted in a Mulvany wire myograph and bathed in Krebs-Henseleit buffer gassed with 95% O(2)/5% CO(2) and maintained at 37 degrees C. Tissues were pre-contracted with the thromboxane mimetic U46619, endothelin-1 or KCl. Cumulative concentration-response curves to beta-adrenoceptor agonists or forskolin were then carried out in the absence or presence of the mitogen-activated protein kinase kinase (MEK) inhibitors PD98059 (10 or 50 microM) or U0126 (10 microM).Key Results: PD98059 caused a concentration-dependent leftward shift in response to isoprenaline (pEC(50) control, 7.5 +/- 0.1; 50 microM PD98059, 8.1 +/- 0.1: P < 0.05). Inhibition of MEK also enhanced the maximum relaxation seen with salbutamol, but not the responses to the beta(1)-adrenoceptor selective agonist xamoterol or the AC activator forskolin. There was no enhancement of the relaxations to beta-adrenoceptor agonists after inhibition of ERK activation in tissues pre-contracted with KCl or treated with the K(+) channel blocker tetraethylammonium.Conclusions and Implications: These data indicate that ERK inhibits beta(2)-adrenoceptor-mediated vasodilatation through a mechanism which may involve inactivation of plasma membrane potassium channels. [ABSTRACT FROM AUTHOR]

In a 2001 report, the U.S. National Institutes of Health called for more integration of the social sciences into health-related research, including research guided by theories and methods that take social and cultural systems into consideration. Based on a theoretical framework that integrates Hofstede's cultural dimensions with sociological theory, the authors used multilevel modeling to explore the association of culture with structural inequality and health disparities. Their results support the idea that cultural dimensions and social structure, along with economic development, may account for much of the cross-national variation in the distribution of health inequalities. Sensitivity tests also suggest that an interaction between culture and social structure may confound the relationship between income inequality and health. It is necessary to identify important cultural and social structural characteristics before we can achieve an understanding of the complex, dynamic systems that affect health, and develop culturally sensitive interventions and policies. This study takes a step toward identifying some of the relevant cultural and structural influences. More research is needed to explore the pathways leading from the sociocultural environment to health inequalities. [ABSTRACT FROM AUTHOR]

Of 490 middle and high school students attending alternative schools who participated in this study, 18.2% of females and 6.7% of males reported a past unwanted sexual experience. Relative to nonabused subjects, female and male adolescents who reported a forced sexual experience were more likely to exhibit externalizing problems such as sexual risk behaviors and internalizing problems such as depression and suicidal thoughts in the past two weeks. Females with a history of a coercive sexual experience also had increased odds of drug and alcohol use, and violence-related behaviors. The magnitude of these associations ranged from a twofold increase to a tenfold increase. Controlling for ethnicity, family income, and family support had less impact on the strength of associations between a forced sexual experience and sexual risk behaviors for females than for males. A forced sexual experience remained significantly associated with depression for females and suicidal thoughts for males. Alcohol use in the past 30 days was the only substance use-related problem that remained significant for females. Violence-related behaviors did not remain significant in the presence of control variables for females. Interventions focusing on adolescents with a history of a coercive sexual experience in general, and in alternative schools in particular, are suggested by these results. More research focusing on the differential impact of a coercive sexual experience on males and females is crucial. [ABSTRACT FROM AUTHOR]

Abstract Background Plasmodium falciparum malaria is a significant problem around the world today, thus there is still a need for new control methods to be developed. Because the sporozoite displays dual infectivity for both the mosquito salivary glands and vertebrate host tissue, it is a good target for vaccine development. Methods The P. falciparum gene, PF11_0394, was chosen as a candidate for study due to its potential role in the invasion of host tissues. This gene, which was selected using a data mining approach from PlasmoDB, is expressed both at the transcriptional and protein levels in sporozoites and likely encodes a putative surface protein. Using reverse transcription-polymerase chain reaction (RT-PCR) and green fluorescent protein (GFP)-trafficking studies, a transcript and protein expression profile of PF11_0394 was determined. Results The PF11_0394 protein has orthologs in other Plasmodium species and Apicomplexans, but none outside of the group Apicomplexa. PF11_0394 transcript was found to be present during both the sporozoite and erythrocytic stages of the parasite life cycle, but no transcript was detected during axenic exoerythrocytic stages. Despite the presence of transcript throughout several life cycle stages, the PF11_0394 protein was only detected in salivary gland sporozoites. Conclusions PF11_0394 appears to be a protein uniquely detected in salivary gland sporozoites. Even though a specific function of PF11_0394 has not been determined in P. falciparum biology, it could be another candidate for a new vaccine.

Abstract Background Although neighbourhood environments are often blamed for contributing to rising levels of obesity, current evidence is based predominantly on cross-sectional samples. This study examined associations between objectively-measured environmental characteristics of neighbourhoods and adiposity cross-sectionally and longitudinally over three years in children and their female carers. Methods Longitudinal study of 140 5-6 year-old and 269 10-12 year-old children and their female carers (n = 369). At baseline (2001) and follow-up (2004), height and weight were measured among children and self-reported among female carers, and were used to compute BMI z-scores and BMI, respectively. A Geographic Information System determined access to destinations (public open spaces, sports options, walking/cycling tracks), road connectivity (density of cul-de-sacs and intersections, proportion of 4-way intersections, length of 'access' paths (overpasses, access lanes, throughways between buildings)) and traffic exposure (length of 'busy' and 'local' roads) within 800 m and 2 km of home. Univariate and multivariable linear regression analyses examined associations between environmental characteristics and BMI/BMI z-scores at baseline and change in BMI/BMI z-scores over the three years. Results Cross-sectionally, BMI z-score was inversely associated with length (km) of access paths within 800 m (b = -0.50) and 2 km (b = -0.16) among younger and number of sport/recreation public open spaces (b = -0.14) and length (km) of 'access' paths (b = -0.94) within 800 m and length of local roads within 2 km (b = -0.01) among older children. Among female carers, BMI was associated with length (km) of walking/cycling tracks (b = 0.17) and busy roads (b = -0.34) within 800 m. Longitudinally, the proportion of intersections that were 4-way (b = -0.01) within 800 m of home was negatively associated with change in BMI z-score among younger children, while length (km) of access paths (b = 0.18) within 800 m was significant among older children. Among female carers, options for aerobics/fitness and swimming within 2 km were associated with change in BMI (B = -0.42). Conclusion A small number of neighbourhood environment features were associated with adiposity outcomes. These differed by age group and neighbourhood scale (800 m and 2 km) and were inconsistent between cross-sectional and longitudinal findings. However, the results suggest that improvements to road connectivity and slowing traffic and provision of facilities for leisure activities popular among women may support obesity prevention efforts.

Abstract Background Recent studies have implicated the epidermal growth factor receptor (EGFR) within the subparaventricular zone as being a major mediator of locomotor and masking behaviors in mice. The results were based on small cohorts of mice homozygous for the hypomorphic Egfrwa2 allele on a mixed, genetically uncontrolled background, and on intraventricular infusion of exogenous EGFR ligands. Subsequenlty, a larger study using the same genetically mixed background failed to replicate the original findings. Since both previous approaches were susceptible to experimental artifacts related to an uncontrolled genetic background, we analyzed the locomotor behaviors in Egfrwa2 mutant mice on genetically defined, congenic backgrounds. Methods Mice carrying the Egfrwa2 hypomorphic allele were bred to congenicity by backcrossing greater than ten generations onto C57BL/6J and 129S1/SvImJ genetic backgrounds. Homozygous Egfrwa2 mutant and wildtype littermates were evaluated for defects in locomotor and masking behaviors. Results Mice homozygous for Egfrwa2 showed normal daily locomotor activity and masking indistinguishable from wildtype littermates at two light intensities (200–300 lux and 400–500 lux). Conclusion Our results demonstrate that reduced EGFR activity alone is insufficient to perturb locomotor and masking behaviors in mice. Our results also suggest that other uncontrolled genetic or environmental parameters confounded previous experiments linking EGFR activity to daily locomotor activity and provide a cautionary tale for genetically uncontrolled studies.

Data on symptoms of major depressive episodes were examined for the 1994 cohort (n = 2,417) of the Alameda County Study (mean age = 65). In addition to age, we examined gender, education, marital status, social isolation and social support, perceived physical and mental health, chronic medical conditions, functional impairment, life events, financial strain, and neighborhood quality. The point prevalence of symptoms of major depressive episodes was 6.6 percent for men and 10.1 percent for women, with a trend for prevalence to increase with age. When the effects of the other psychosocial risk factors were controlled, there were no significant age effects. Multivariate analyses demonstrated that apparent initial age effects were due almost entirely to chronic health problems and functional impairment. The implications are clear: Healthy, normal-functioning older adults are at no greater risk of depression than younger adults. Apparent age-related effects on depression are attributable to physical health problems and related disability. [ABSTRACT FROM AUTHOR]

Although religiosity is protective for mortality and morbidity, its relationship with depression is unclear. We used the 1994 Alameda County Study survey of 2,537 subjects aged 50-102 to analyze associations between two forms of religiosity and depression as well as the extent to which religiosity buffers relationships between stressors and depression. Non-organizational religiosity included prayer and importance of religious and spiritual beliefs; organizational religiosity included attendance at services and other activities. Non-organizational religiosity had no association with depression; organizational religiosity had a negative relationship that weakened slightly with the addition of health controls. Both forms of religiosity buffered associations with depression for non-family stressors, such as financial and health problems. However, non-organizational religiosity exacerbated associations with depression for child problems, and organizational religiosity exacerbated associations with depression for marital problems, abuse, and caregiving. Religiosity may help those experiencing non-family stressors, but may worsen matters for those facing family crises. [ABSTRACT FROM AUTHOR]

Limited data on ethnic group differences among young adolescents exist regarding the prevalence of mental distress, social stress, and resources. The aim of this cross-sectional study was to examine ethnic differences among African American (AA), European American (EA), Hispanic American (HA), and Asian American adolescents in mental distress, social stress, and resources. A total of 316 adolescents completed the self-administered questionnaire: The DSD, Somatic Symptom Scale, SAFE-C, FES, Coping, and Rosenberg's Self-Esteem Scale. The findings revealed that (1) Asian Americans and HAs reported significantly higher scores on social stress and mental distress and lower scores on resources than EAs; (2) Compared to EAs, AAs and HAs were more likely to experience social stress (2.2 < or = OR < or = 4.3), and HAs are more likely to have suicidal ideation (OR = 2.04; 95% CI = 1.04-3.98). Findings highlighted the importance of understanding the role of social stress in the development of adolescents' mental distress. Identified ethnic minority adolescents are vulnerable groups who require additional research and attention from health care providers. [ABSTRACT FROM AUTHOR]

The goal of this study was to explore whether a history of sexual abuse is associated with high-risk sexual behaviors among female adolescents attending alternative schools in a large urban city in the southwestern United States, and to examine the role of depression and substance abuse in explaining this association. One hundred eighty-four sexually active female adolescents constituted the sample for this analysis. Forty-nine (26.6%) reported that they were forced to have sex. Having a history of sexual abuse substantially increased sexual risk behaviors. Adolescents reporting a history of sexual abuse, compared to those who did not report such a history, were significantly more likely to have initiated sexual activity (intercourse) before age 14, to have had three or more sexual partners in the last 3 months, and to have had a history of sexually transmitted diseases. These associations remained significant afar controlling for age, ethnicity, and family income. Depression and substance abuse did not explain the association between sexual abuse and high-risk sexual behaviors. It seems reasonable to conclude that adolescents with a history of sexual abuse have greater difficulty practicing safe sexual behaviors than do those who have not been sexually abused. Given the prevalence of child sexual abuse and the extent of its impact, it is critical that intervention strategies for adolescent females address the issue of abuse and help them adopt self-protective sexual behaviors. The findings also highlight the importance of targeting adolescents who attend alternative schools. [ABSTRACT FROM AUTHOR]

Chemical communication using pheromones is thought to have contributed to the diversification and speciation of insects. The species-specific pheromones are detected by specialized pheromone receptors (PRs). Whereas the evolution and function of PRs have been extensively studied in Lepidoptera, only a few PRs have been identified in beetles, which limits our understanding of their evolutionary histories and physiological functions. To shed light on these questions, we aimed to functionally characterize potential PRs in the spruce bark beetle Ips typographus ("Ityp") and explore their evolutionary origins and molecular interactions with ligands. Males of this species release an aggregation pheromone comprising 2-methyl-3-buten-2-ol and (4S)-cis-verbenol, which attracts both sexes to attacked trees. Using two systems for functional characterization, we show that the highly expressed odorant receptor (OR) ItypOR41 responds specifically to (4S)-cis-verbenol, with structurally similar compounds eliciting minor responses. We next targeted the closely related ItypOR40 and ItypOR45. Whereas ItypOR40 was unresponsive, ItypOR45 showed an overlapping response profile with ItypOR41, but a broader tuning. Our phylogenetic analysis shows that these ORs are present in a different OR clade as compared to all other known beetle PRs, suggesting multiple evolutionary origins of PRs in bark beetles. Next, using computational analyses and experimental validation, we reveal two amino acid residues (Gln179 and Trp310) that are important for ligand binding and pheromone specificity of ItypOR41 for (4S)-cis-verbenol, possibly via hydrogen bonding to Gln179. Collectively, our results shed new light on the origins, specificity, and ligand binding mechanisms of PRs in beetles., Competing Interests: Conflict of Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

Identification of youth presenting early risk factors for psychosis may facilitate preventive intervention. Through school-based screening, we recruited 112 children aged 9-12 years who presented multiple putative antecedents of schizophrenia (ASz), a family history of schizophrenia (FHx), or neither of these risk factors (typically-developing; TD). Clinical and functional outcomes were assessed at age 17-21 years (N = 93). Compared to the TD group, the ASz group had higher total Prodromal Questionnaire (PQ) scores (β = 10.59, 95% CI = 3.76, 17.42) and total psychopathology scores (β = 6.13, 95% CI: 1.03, 11.23), were more likely to score above-threshold on the PQ positive symptoms scale (OR = 4.00, 95% CI = 1.08, 14.83), and had lower scores on the Social and Occupational Functioning Scale (β = -9.43, 95% CI = -15.08, -3.77) at follow-up. The FHx and TD groups did not differ on any outcome. Findings suggest that population screening for putative antecedents of schizophrenia may identify children who would benefit from preventative intervention., (© 2024. The Author(s).)

Monocytes and macrophages (Mos/Mϕs) play diverse roles in wound healing by adopting a spectrum of functional phenotypes; however, the regulation of such heterogeneity remains poorly defined. We enhanced our previously published Bayesian inference TF activity model, incorporating both single-cell RNA sequencing and single-cell ATAC sequencing data to infer transcription factor (TF) activity in Mos/Mϕs during skin wound healing. We found that wound Mos/Mϕs clustered into early-stage Mos/Mϕs, late-stage Mϕs, and APCs, and that each cluster showed differential chromatin accessibility and differential predicted TF activity that did not always correlate with mRNA or protein expression. Network analysis revealed two highly connected large communities involving a total of 19 TFs, highlighting TF cooperation in regulating wound Mos/Mϕs. This analysis also revealed a small community populated by NR4A1 and NFKB1, supporting a proinflammatory link between these TFs. Importantly, we validated a proinflammatory role for NR4A1 activity during wound healing, showing that Nr4a1 knockout mice exhibit decreased inflammatory gene expression in early-stage wound Mos/Mϕs, along with delayed wound re-epithelialization and impaired granulation tissue formation. In summary, our study provides insight into TF activity that regulates Mo/Mϕ heterogeneity during wound healing and provides a rational basis for targeting Mo/Mϕ TF networks to alter phenotypes and improve healing., (Copyright © 2024 by The American Association of Immunologists, Inc.)

Chronic wounds in diabetic patients are associated with significant morbidity and mortality; however, few therapies are available to improve healing of diabetic wounds. Our group previously reported that low-intensity vibration (LIV) could improve angiogenesis and wound healing in diabetic mice. The purpose of this study was to begin to elucidate the mechanisms underlying LIV-enhanced healing. We first demonstrate that LIV-enhanced wound healing in db/db mice is associated with increased IGF1 protein levels in liver, blood, and wounds. The increase in insulin-like growth factor (IGF) 1 protein in wounds is associated with increased Igf1 mRNA expression both in liver and wounds, but the increase in protein levels preceded the increase in mRNA expression in wounds. Since our previous study demonstrated that liver was a primary source of IGF1 in skin wounds, we used inducible ablation of IGF1 in the liver of high-fat diet (HFD)-fed mice to determine whether liver IGF1 mediated the effects of LIV on wound healing. We demonstrate that knockdown of IGF1 in liver blunts LIV-induced improvements in wound healing in HFD-fed mice, particularly increased angiogenesis and granulation tissue formation, and inhibits the resolution of inflammation. This and our previous studies indicate that LIV may promote skin wound healing at least in part via crosstalk between the liver and wound. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland., (© 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.)

Insects are able to detect a plethora of olfactory cues using a divergent family of odorant receptors (ORs). Despite the divergent nature of this family, related species frequently express several evolutionarily conserved OR orthologues. In the largest order of insects, Coleoptera, it remains unknown whether OR orthologues have conserved or divergent functions in different species. Using HEK293 cells, we addressed this question through functional characterization of two groups of OR orthologues in three species of the Curculionidae (weevil) family, the conifer-feeding bark beetles Ips typographus L. ("Ityp") and Dendroctonus ponderosae Hopkins ("Dpon") (Scolytinae), and the pine weevil Hylobius abietis L. ("Habi"; Molytinae). The ORs of H. abietis were annotated from antennal transcriptomes. The results show highly conserved response specificities, with one group of orthologues (HabiOR3/DponOR8/ItypOR6) responding exclusively to 2-phenylethanol (2-PE), and the other group (HabiOR4/DponOR9/ItypOR5) responding to angiosperm green leaf volatiles (GLVs). Both groups of orthologues belong to the coleopteran OR subfamily 2B, and share a common ancestor with OR5 in the cerambycid Megacyllene caryae, also tuned to 2-PE, suggesting a shared evolutionary history of 2-PE receptors across two beetle superfamilies. The detected compounds are ecologically relevant for conifer-feeding curculionids, and are probably linked to fitness, with GLVs being used to avoid angiosperm nonhost plants, and 2-PE being important for intraspecific communication and/or playing a putative role in beetle-microbe symbioses. To our knowledge, this study is the first to reveal evolutionary conservation of OR functions across several beetle species and hence sheds new light on the functional evolution of insect ORs., (© 2022 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Background: In individuals at clinical high-risk for psychosis, elevated cortisol levels predict subsequent onset of psychotic disorder. However, it is unclear whether cortisol alterations are evident at an earlier clinical stage and promote progression of psychosis expression. This study aimed to address this issue by investigating whether cortisol levels in childhood were associated with the emergence of attenuated psychotic symptoms in early adulthood. In exploratory analyses, we examined whether cortisol and psychosocial stress measures interacted in predicting attenuated psychotic symptoms., Methods: A sample of children (N = 109) enriched for psychosis risk factors were recruited at age 9-12 years and assessed at age 11-14 years (T1) and 17-21 years (T2). Measures of psychopathology, psychosocial stressors, and salivary cortisol were obtained at T1. Attenuated psychotic symptoms were assessed at T2 using the Prodromal Questionnaire., Results: Diurnal cortisol (β = 0.915, 95% CI: 0.062-1.769) and daily stressors (β = 0.379, 95% CI: 0.034-0.723) at T1 were independently associated with total Prodromal Questionnaire scores at T2 after accounting for demographic factors and T1 psychopathology. Exploratory analyses indicated a significant interaction between T1 diurnal cortisol and daily stressors (β = 0.743, 95% CI: 0.081-1.405), with the highest predicted T2 total Prodromal Questionnaire scores occurring when both diurnal cortisol and daily stressors were increased., Conclusions: Our findings suggest that daily stressors and elevations in diurnal cortisol in late childhood/early adolescence increases risk for developing attenuated psychotic symptoms. These findings emphasize the importance of assessing environmental and biological risk factors for psychosis during neurodevelopmentally vulnerable time periods., (Copyright © 2021 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

Insulin-like growth factor (IGF)-1 plays important role in tissue repair through its ability to stimulate wound cell activity. While IGF-1 is expressed locally by wound cells, liver-derived IGF-1 is also present at high levels in the circulation, and the contributions of local vs circulating IGF-1 to wound levels remain undefined. The hypothesis of this study was that liver is a primary source of IGF-1 during skin wound healing. To test this hypothesis, we utilized a model that allows inducible ablation of IGF-1 specifically in liver of adult mice. We demonstrate that ablation of liver IGF-1 leads to >85% loss of circulating IGF-1 and ~60% decrease in wound IGF-1 during the proliferative phase of healing in both male and female mice. This reduction of liver-derived IGF-1 did not alter local mRNA expression of Igf1 in wounds. Knockdown of liver IGF-1 significantly delayed wound re-epithelialization and reduced granulation tissue formation and collagen deposition. Knockdown of liver IGF-1 also significantly reduced angiogenesis and resulted in persistent macrophage accumulation. In summary, liver is a primary source of IGF-1 in skin wounds and contributes to many aspects of both epithelial and dermal healing.

Insects detect odors using an array of odorant receptors (ORs), which may expand through gene duplication. How and which new functions may evolve among related ORs within a species remain poorly investigated. We addressed this question by functionally characterizing ORs from the Eurasian spruce bark beetle Ips typographus, in which physiological and behavioral responses to pheromones, volatiles from host and nonhost trees, and fungal symbionts are well described. In contrast, knowledge of OR function is restricted to two receptors detecting the pheromone compounds (S)-(-)-ipsenol (ItypOR46) and (R)-(-)-ipsdienol (ItypOR49). These receptors belong to an Ips-specific OR-lineage comprising seven ItypORs. To gain insight into the functional evolution of related ORs, we characterized the five remaining ORs in this clade using Xenopus oocytes. Two receptors responded primarily to the host tree monoterpenes (+)-3-carene (ItypOR25) and p-cymene (ItypOR27). Two receptors responded to oxygenated monoterpenoids produced in larger relative amounts by the beetle-associated fungi, with ItypOR23 specific for (+)-trans-(1R, 4S)-4-thujanol, and ItypOR29 responding to (+)-isopinocamphone and similar ketones. ItypOR28 responded to the pheromone E-myrcenol from the competitor Ips duplicatus. Overall, the OR responses match well with those of previously characterized olfactory sensory neuron classes except that neurons detecting E-myrcenol have not been identified. The characterized ORs are under strong purifying selection and demonstrate a shared functional property in that they all primarily respond to monoterpenoids. The variation in functional groups among OR ligands and their diverse ecological origins suggest that neofunctionalization has occurred early in the evolution of this OR-lineage following gene duplication., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Insect odorant receptor (OR) genes are routinely expressed in Human Embryonic Kidney (HEK) 293 cells for functional characterization ("de-orphanization") using transient or stable expression. However, progress in this research field has been hampered because some insect ORs are not functional in this system, which may be due to insufficient protein levels. We investigated whether codon optimization of insect OR sequences for expression in human cells could facilitate their functional characterization in HEK293 cells with stable and inducible expression. We tested the olfactory receptor co-receptor (Orco) proteins from the bark beetles Ips typographus ("Ityp") and Dendroctonus ponderosae ("Dpon"), and six ItypORs previously characterized in Xenopus laevis oocytes and/or HEK cells. Western blot analysis indicated that codon optimization yielded increased cellular protein levels for seven of the eight receptors. Our experimental assays demonstrated that codon optimization enabled functional characterization of two ORs (ItypOR25 and ItypOR29) which are unresponsive when expressed from wildtype (non-codon optimized) genes. Similar to previous Xenopus oocyte recordings, ItypOR25 responded primarily to the host/conifer monoterpene (+)-3-carene. ItypOR29 responded primarily to (+)-isopinochamphone and similar ketones produced by fungal symbionts and trees. Codon optimization also resulted in significantly increased responses in ItypOR49 to its pheromone ligand ( R )-(-)-ipsdienol, and improved responses to the Orco agonist VUAA1 in ItypOrco. However, codon optimization did not result in functional expression of DponOrco, ItypOR23, ItypOR27, and ItypOR28 despite higher protein levels as indicated by Western blots. We conclude that codon optimization may enable or improve the functional characterization of insect ORs in HEK cells, although this method is not sufficient for all ORs that are not functionally expressed from wildtype genes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Roberts, Yuvaraj and Andersson.)

Ethnopharmacological Relevance: Asphodelus tenuifolius Cav. (Asphodelaceae), a wild, terrestrial, annual stemless herb, is widely used in traditional medicine for the treatment of hypertension, diabetes, atherosclerosis and circulatory problems. A previous research study from our laboratory revealed that A. tenuifolius has beneficial effects in reducing blood pressure and improves aortic endothelial dysfunction in chronically glucose fed rats. Despite the fact that A. tenuifolius reduces blood pressure and improves endothelial function in vivo, there are no detailed studies about its possible mechanism of action., Aim of the Study: This study was designed to provide pharmacological basis and mechanism of action for the traditional use of A. tenuifolius in hypertension and circulatory problems. We explored the vasorelaxant effect of A. tenuifolius and its underlying vasorelaxation mechanism in porcine coronary artery rings., Materials and Methods: Aqueous methanolic crude extract of A. tenuifolius was prepared by maceration process and then activity guided fractionation was carried out by using different polarity based solvents. Phytochemical studies were carried out using LC-DAD-MS. Segments of porcine distal coronary artery were set up in a wire myograph for isometric force measurements. Extract/fractions of A. tenuifolius seeds were tested for vasodilator activity by measurement of changes in tone after pre-contraction with the thromboxane mimetic U46619 in the presence or absence of inhibitors of intracellular signaling cascades., Results: Crude extract/fractions of A. tenuifolius produced dose dependent endothelium independent vasorelaxant response in coronary rings, whereas, the butanol fraction of A. tenuifolius (BS-AT) produced the largest relaxation response with 100% relaxation at 1 mg/ml, therefore the mechanism of relaxation of this fraction was determined. The relaxation to BS-AT was unaffected by removal of the endothelium, pre-contraction with KCl, or the presence of the non-selective potassium channel blocker tetraethylammonium, indicating that the relaxation was endothelium-independent, and does not involve activation of potassium channels. BS-AT (1 mg/ml) inhibited the contractile response to calcium,the L-type calcium channel activator BAY K8664,and ionomycin, indicating that it inhibits calcium-induced contractions. The relaxation response to BS-AT was attenuated in the absence of extracellular calcium. However, relaxations to BS-AT were also reduced after deletion of calcium from intracellular stores with cyclopiazonic acid. Incubation with 1 mg/ml BS-AT also inhibited phosphorylation of myosin light chains in homogenates of coronary artery., Conclusion: The butanol extract of Asphodelus tenuifolius produces a large endothelium-independent relaxation of the porcine coronary artery through inhibition of calcium-induced contractions. The effect appears to be downstream of calcium influx, possibly through inhibition of myosin light chain kinase. This study supports previous studies demonstrating that A. tenuifolius reduces blood pressure. Future studies will aim to determine the active compounds underlying this response., (Copyright © 2020. Published by Elsevier B.V.)

In order to gain insight into the regulation of vascular tone by mitochondria, the effects of mitochondrial complex III inhibitors on contractile responses in porcine isolated coronary arteries were investigated. Segments of porcine coronary arteries were set up for isometric tension recording and concentration response curves to contractile agents were carried out in the absence or presence of the complex III inhibitors antimycin A or myxothiazol. Activity of AMP kinase was determined by measuring changes in phosphorylation of AMP kinase at Thr172. Pre-incubation with 10 μM antimycin A (Q i site inhibitor), or myxothiazol (Q o site inhibitor) led to inhibition of the contraction to the thromboxane receptor agonist U46619. Similar effects were seen on contractile responses to extracellular calcium, and the L-type calcium channel opener BAY K 8644, suggesting that both antimycin A and myxothiazol inhibit calcium-dependent contractions. The inhibitory effect of antimycin A was still seen in the absence of extracellular calcium, indicating an additional effect on a calcium independent pathway. The AMP kinase inhibitor dorsomorphin (10 μM) prevented the inhibitory of antimycin A but not myxothiazol. Furthermore, antimycin A increased the phosphorylation of AMP kinase, indicating an increase in activity, suggesting that antimycin A also acts through this pathway. These data indicate that inhibition of complex III attenuates contractile responses through inhibition of calcium influx. However, inhibition of the Q i site can also inhibit the contractile response through activation of AMP kinase., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Chronic wounds in diabetic patients represent an escalating health problem, leading to significant morbidity and mortality. Our group previously reported that whole body low-intensity vibration (LIV) can improve angiogenesis and wound healing in diabetic mice. The purpose of the current study was to determine whether effects of LIV on wound healing are frequency and/or amplitude dependent. Wound healing was assessed in diabetic (db/db) mice exposed to one of four LIV protocols with different combinations of two acceleration magnitudes (0.3 and 0.6 g) and two frequencies (45 and 90 Hz) or in non-vibration sham controls. The low acceleration, low frequency protocol (0.3 g and 45 Hz) was the only one that improved wound healing, increasing angiogenesis and granulation tissue formation, leading to accelerated re-epithelialization and wound closure. Other protocols had little to no impact on healing with some evidence that 0.6 g accelerations negatively affected wound closure. The 0.3 g, 45 Hz protocol also increased levels of insulin-like growth factor-1 and tended to increase levels of vascular endothelial growth factor in wounds, but had no effect on levels of basic fibroblast growth factor or platelet derived growth factor-bb, indicating that this LIV protocol induces specific growth factors during wound healing. Our findings demonstrate parameter-dependent effects of LIV for improving wound healing that can be exploited for future mechanistic and therapeutic studies., Competing Interests: TK has a patent pending on the application of vibration for therapeutic treatment for tissue repair “Method and system for physical stimulation of tissue” (#2013/0165,824). The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Roberts, Bilgen, Kineman and Koh.)

Background: Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize ecologically important ORs, using two systems for heterologous expression. Our final aim was to gain insight into the ligand-OR interaction of the functionally characterized ORs, using a combination of computational and experimental methods., Results: We annotated 73 ORs from an antennal transcriptome of I. typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are responsive to single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. Their responses and antennal expression correlate with the specificities, localizations, and/or abundances of olfactory sensory neurons detecting these enantiomers. We use homology modeling and molecular docking to predict their binding sites. Our models reveal a likely binding cleft lined with residues that previously have been shown to affect the responses of insect ORs. Within this cleft, the active ligands are predicted to specifically interact with residues Tyr84 and Thr205 in ItypOR46. The suggested importance of these residues in the activation by ipsenol is experimentally supported through site-directed mutagenesis and functional testing, and hydrogen bonding appears key in pheromone binding., Conclusions: The emerging insight into ligand binding in the two characterized ItypORs has a general importance for our understanding of the molecular and functional evolution of the insect OR gene family. Due to the ecological importance of the characterized receptors and widespread use of ipsenol and ipsdienol in bark beetle chemical communication, these ORs should be evaluated for their potential use in pest control and biosensors to detect bark beetle infestations.

Background: Mismatch negativity (MMN) and P3a amplitude reductions are robust abnormalities of sensory information processing in schizophrenia, but they are variably present in different profiles of risk (family history vs. clinical high risk) for the disorder. This study aimed to determine whether these abnormalities characterize children presenting replicated risk factors for schizophrenia, using longitudinal assessment over the ages of 9-16 years in children with multiple replicated antecedents of schizophrenia (ASz) and with family history of schizophrenia (FHx), relative to typically developing (TD) peers., Methods: A total of 105 children (52 female) sampled from the community were assessed at ages 9-12 years and approximately 2 and 4 years later. Linear mixed models were fitted to MMN and P3a peak amplitudes and latencies, with intercept and slope estimates from 32 ASz and 28 FHx children compared with those of 45 TD peers., Results: In ASz relative to TD children, MMN amplitude initially increased and then prominently decreased during adolescence. Both ASz and FHx children had greater P3a amplitude than TD children at 11 years, which decreased with age, in contrast to P3a amplitude increasing during adolescence in TD youths. MMN abnormalities were specific to ASz children who continued to present symptoms during follow-up., Conclusions: Age-dependent MMN and P3a abnormalities demarcate adolescent development of ASz and FHx from TD children, with auditory change detection abnormalities specific to ASz children with continuing symptoms and attention-orienting abnormalities characterizing both ASz and FHx risk profiles. Follow-up is required to determine whether these abnormalities index vulnerability for schizophrenia or an illness nonspecific developmental delay., (Copyright © 2020 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

The construction of a low affinity Ca 2+ -probe that locates to the cell cortex and cell surface wrinkles, is described called. EPIC3 (ezrin-protein indicator of Ca 2+ ). The novel probe is a fusion of CEPIA3 with ezrin, and is used in combination with a Ca 2+ -insensitive probe, ezrin-mCherry, both of which locate at the cell cortex. EPIC3 was used to monitor the effect of Ca 2+ influx on intra-wrinkle Ca 2+ in the macrophage cell line, RAW 264.7. During experimentally-induced Ca 2+ influx, EPIC3 reported Ca 2+ concentrations at the cell cortex in the region of 30-50 μM, with peak locations towards the tips of wrinkles reaching 80 μM. These concentrations were associated with cleavage of ezrin (a substrate for the Ca 2+ activated protease calpain-1) and released the C-terminal fluors. The cortical Ca 2+ levels, restricted to near the site of phagocytic cup formation and pseudopodia extension during phagocytosis also reached high levels (50-80 μM) during phagocytosis. As phagocytosis was completed, hotspots of Ca 2+ near the phagosome were also observed., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hypothalamic amenorrhoea (HA) accounts for approximately 30% of cases of secondary amenorrhoea in women of reproductive age. It is caused by deficient secretion of hypothalamic gonadotrophin-releasing hormone, which in turn leads to failure of pituitary gonadotrophin and gonadal steroid release. Functional HA (FHA) is defined as HA occurring in the absence of a structural lesion and is predominantly caused by significant weight loss, intense exercise or stress. Treatment of FHA is crucial in avoiding the long-term health consequences on fertility and bone health, in addition to reducing psychological morbidity. This article summarises our understanding of the mechanisms underlying FHA, the evidence base for its clinical management and emerging therapies., Competing Interests: Conflict of interest statement: The authors declare that there is no conflict of interest., (© The Author(s), 2020.)

The intracellular events underlying phagocytosis, a crucial event for innate immunity, are still unresolved. In order to test whether the reservoir of membrane required for the formation of the phagocytic pseudopodia is maintained by cortical ezrin, and that its cleavage is a key step in releasing this membrane, the cleavage of cortical ezrin was monitored within living phagocytes (the phagocytically competent cell line RAW264.7) through expressing two ezrin constructs with fluorescent protein tags located either inside the FERM or at the actin-binding domains. When ezrin is cleaved in the linker region by the Ca 2+ -activated protease calpain, separation of the two fluorophores would result. Experimentally induced Ca 2+ influx triggered cleavage of peripherally located ezrin, which was temporally associated with cell expansion. Ezrin cleavage was also observed in the phagocytic pseudopodia during phagocytosis. Thus, our data demonstrates that peripheral ezrin is cleaved during Ca 2+ -influx-induced membrane expansion and locally within the extending pseudopodia during phagocytosis. This is consistent with a role for intact ezrin in maintaining folded membrane on the cell surface, which then becomes available for cell spreading and phagocytosis., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

This article investigates the effect of using different playing speeds of 2D animation on the decision accuracy and reaction times of elite footballers. Groups of players from different age categories (under 16, under 18 and under 23 year olds) elite academy footballers watched 2D animations at five different ratios of real-life speed (0.5, 0.75, 1, 1.25 and 1.5 times real-world speed). Participants responded by choosing, from a number of options, which was the best attacking option in each 2D animation clip. After each clip, players also rated each clip for how "game-like" or realistic the scenario felt. Results showed a significant difference between all groups, whereby participants became faster and more accurate by age. Also, the result of "game-like" perception probing which compared perception in each group to each speed showed that the under 16 age group had statistically lower perception accuracy compared to the under 18 and under 23 age groups. This highlights that the more expertise a player has the faster information is processed., (© 2020 Elsevier B.V. All rights reserved.)

The measurement and manipulation of cytosolic free Ca 2+ of neutrophils is crucial for investigating the mechanisms within living neutrophils which generate Ca 2+ signals and the cellular responses triggered by them. Optical methods for this are the most applicable for neutrophils, and are discussed here, especially the use of fluorescent indicators of Ca 2+ and photoactivation of reagents involved in Ca 2+ signaling. Both of these synthetic agents can be loaded into neutrophils as lipid-soluble esters or can be microinjected into the cell. In this chapter, we outline some of the techniques that have been used to monitor, visualize, and manipulate Ca 2+ in neutrophils.