A Cataract-Causing Mutation in the TRPM3 Cation Channel Disrupts Calcium Dynamics in the Lens.

TRPM3 belongs to the melastatin sub-family of transient receptor potential (TRPM) cation channels and has been shown to function as a steroid-activated, heat-sensitive calcium ion (Ca2+) channel. A missense substitution (p.I65M) in the TRPM3 gene of humans (TRPM3) and mice (Trpm3) has been shown to underlie an inherited form of early-onset, progressive cataract. Here, we model the pathogenetic effects of this cataract-causing mutation using 'knock-in' mutant mice and human cell lines. Trpm3 and its intron-hosted micro-RNA gene (Mir204) were strongly co-expressed in the lens epithelium and other non-pigmented and pigmented ocular epithelia. Homozygous Trpm3-mutant lenses displayed elevated cytosolic Ca2+ levels and an imbalance of sodium (Na+) and potassium (K+) ions coupled with increased water content. Homozygous TRPM3-mutant human lens epithelial (HLE-B3) cell lines and Trpm3-mutant lenses exhibited increased levels of phosphorylated mitogen-activated protein kinase 1/extracellular signal-regulated kinase 2 (MAPK1/ERK2/p42) and MAPK3/ERK1/p44. Mutant TRPM3-M65 channels displayed an increased sensitivity to external Ca2+ concentration and an altered dose response to pregnenolone sulfate (PS) activation. Trpm3-mutant lenses shared the downregulation of genes involved in insulin/peptide secretion and the upregulation of genes involved in Ca2+ dynamics. By contrast, Trpm3-deficient lenses did not replicate the pathophysiological changes observed in Trpm3-mutant lenses. Collectively, our data suggest that a cataract-causing substitution in the TRPM3 cation channel elicits a deleterious gain-of-function rather than a loss-of-function mechanism in the lens.

Cornual pregnancy rupture and massive hemorrhage: A case report.

BACKGROUND: Corneal pregnancy is rare and difficult to detect in the early stages. Due to the abundant blood supply in this area, a rupture can result in massive internal bleeding, shock, and even death. Therefore, immediate surgery is necessary, and patients must replenish their blood volume as soon as possible to ensure blood supply to important organs. For those whose blood pressure cannot immediately rise, surgery should be performed while resisting shock to buy time. CASE SUMMARY: We present the case of a 34-year-old Chinese woman at 19 weeks of gestation who had a corneal pregnancy. No abnormalities were detected in the examinations in the first trimester. This patient was 19 weeks pregnant and sought medical advice due to sudden lower abdominal pain, syncope, and hemorrhagic shock. After rescue and treatment, she recovered and was discharged from the hospital, afterwards, the patient gave birth to a child 7 years later. CONCLUSION: The early diagnosis of cornual pregnancy is mainly based on ultrasound. However, there is a high incidence of missed diagnosis and misdiagnosis of this disease. Patients may face serious and life-threatening conditions in case of the rupture of cornual pregnancy. This disease can be mainly treated by surgery.

Charged multivesicular body protein 4b forms complexes with gap junction proteins during lens fiber cell differentiation.

Charged multivesicular body protein 4b (CHMP4B) is a core sub-unit of the endosomal sorting complex required for transport III (ESCRT-III) machinery that serves myriad remodeling and scission processes of biological membranes. Mutation of the human CHMP4B gene underlies rare forms of early-onset lens opacities or cataracts, and CHMP4B is required for lens growth and differentiation in mice. Here, we determine the sub-cellular distribution of CHMP4B in the lens and uncover a novel association with gap junction alpha-3 protein (GJA3) or connexin 46 (Cx46) and GJA8 or Cx50. Immunofluorescence confocal microscopy revealed that CHMP4B localized to cell membranes of elongated fiber cells in the outer cortex of the lens-where large gap junction plaques begin to form-particularly, on the broad faces of these flattened hexagon-like cells in cross-section. Dual immunofluorescence imaging showed that CHMP4B co-localized with gap junction plaques containing Cx46 and/or Cx50. When combined with the in situ proximity ligation assay, immunofluorescence confocal imaging indicated that CHMP4B lay in close physical proximity to Cx46 and Cx50. In Cx46-knockout (Cx46-KO) lenses, CHMP4B-membrane distribution was similar to that of wild-type, whereas, in Cx50-KO lenses, CHMP4B localization to fiber cell membranes was lost. Immunoprecipitation and immunoblotting analyses revealed that CHMP4B formed complexes with Cx46 and Cx50 in vitro. Collectively, our data suggest that CHMP4B forms plasma membrane complexes, either directly and/or indirectly, with gap junction proteins Cx46 and Cx50 that are often associated with "ball-and-socket" double-membrane junctions during lens fiber cell differentiation.

Whole-exome sequencing prioritizes candidate genes for hereditary cataract in the Emory mouse mutant.

The Emory cataract (Em) mouse mutant has long been proposed as an animal model for age-related or senile cataract in humans-a leading cause of visual impairment. However, the genetic defect(s) underlying the autosomal dominant Em phenotype remains elusive. Here, we confirmed development of the cataract phenotype in commercially available Em/J mice [but not ancestral Carworth Farms White (CFW) mice] at 6-8 months of age and undertook whole-exome sequencing of candidate genes for Em. Analysis of coding and splice-site variants did not identify any disease-causing/associated mutations in over 450 genes known to underlie inherited and age-related forms of cataract and other lens disorders in humans and mice, including genes for lens crystallins, membrane/cytoskeleton proteins, DNA/RNA-binding proteins, and those associated with syndromic/systemic forms of cataract. However, we identified three cataract/lens-associated genes each with one novel homozygous variant including predicted missense substitutions in Prx (p.R167C) and Adamts10 (p.P761L) and a disruptive in-frame deletion variant (predicted missense) in Abhd12 (p.L30_A32delinsS) that were absent in CFW and over 35 other mouse strains. In silico analysis predicted that the missense substitutions in Prx and Adamts10 were borderline neutral/damaging and neutral, respectively, at the protein function level, whereas, that in Abhd12 was functionally damaging. Both the human counterparts of Adamts10 and Abhd12 are clinically associated with syndromic forms of cataract known as Weil-Marchesani syndrome 1 and polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataract syndrome, respectively. Overall, while we cannot exclude Prx and Adamts10, our data suggest that Abhd12 is a promising candidate gene for cataract in the Em/J mouse.

"Ick bin een Berlina": dialect proficiency impacts a robot's trustworthiness and competence evaluation.

Background: Robots are increasingly used as interaction partners with humans. Social robots are designed to follow expected behavioral norms when engaging with humans and are available with different voices and even accents. Some studies suggest that people prefer robots to speak in the user's dialect, while others indicate a preference for different dialects. Methods: Our study examined the impact of the Berlin dialect on perceived trustworthiness and competence of a robot. One hundred and twenty German native speakers (M age = 32 years, SD = 12 years) watched an online video featuring a NAO robot speaking either in the Berlin dialect or standard German and assessed its trustworthiness and competence. Results: We found a positive relationship between participants' self-reported Berlin dialect proficiency and trustworthiness in the dialect-speaking robot. Only when controlled for demographic factors, there was a positive association between participants' dialect proficiency, dialect performance and their assessment of robot's competence for the standard German-speaking robot. Participants' age, gender, length of residency in Berlin, and device used to respond also influenced assessments. Finally, the robot's competence positively predicted its trustworthiness. Discussion: Our results inform the design of social robots and emphasize the importance of device control in online experiments.

Effect of Acupoint Hot Compress on Postpartum Urinary Retention After Vaginal Delivery: A Randomized Clinical Trial.

Importance: Acupoint hot compress during the early postpartum period may benefit patients after a vaginal delivery, but the evidence of this effect is limited. Objective: To assess whether acupoint hot compress involving the abdominal, lumbosacral, and plantar regions could reduce the incidence of postpartum urinary retention, relieve postpartum uterine contraction pain, prevent emotional disorders, and promote lactation. Design, Setting, and Participants: This multicenter randomized clinical trial was conducted at 12 hospitals in China. Pregnant patients were screened for eligibility (n = 13 949) and enrolled after vaginal delivery (n = 1200) between January 17 and August 15, 2021; data collection was completed on August 18, 2021. After vaginal delivery, these participants were randomized 1:1 to either the intervention group or control group. Statistical analysis was based on per-protocol population. Interventions: Participants in the control group received routine postpartum care. Participants in the intervention group received routine postpartum care plus 3 sessions of a 4-hour acupoint hot compress involving the abdominal, lumbosacral, and plantar regions within 30 minutes, 24 hours, and 48 hours after delivery. Main Outcomes and Measures: The primary outcome was the incidence of postpartum urinary retention, defined as the first urination occurring more than 6.5 hours after delivery and/or use of an indwelling catheter within 72 hours after delivery. The secondary outcomes were postpartum uterine contraction pain intensity (assessed with the visual analog scale [VAS]), depressive symptoms (assessed with the Edinburgh Postnatal Depression Scale), and lactation conditions (including lactation initiation time, breastfeeding milk volume, feeding mood and times, and newborn weight). Results: Of the 1200 participants randomized, 1085 completed the study (537 in the intervention group and 548 in the control group, with a median [IQR] age of 26.0 [24.0-29.0] years). Participants in the intervention group compared with the control group had significantly decreased incidence of postpartum urinary retention (relative risk [RR], 0.58; 95% CI, 0.35-0.98; P = .03); improved postpartum uterine contraction pain when measured at 6.5 hours (median [IQR] VAS score, 1 [1-2] vs 2 [1-2]; P < .001), 28.5 hours (median [IQR] VAS score, 1 [0-1] vs 1 [1-2]; P < .001), 52.5 hours (median [IQR] VAS score, 1 [0-1] vs 1 [0-1]; P < .001), and 76.5 hours (median [IQR] VAS score, 0 [0-1] vs 0 [0-1]; P = .01) after delivery; reduced depressive symptoms (RR, 0.73; 95% CI, 0.54-0.98; P = .01); and increased breastfeeding milk volume measured at 28.5, 52.5, and 76.5 hours after delivery. No adverse events occurred in either of the 2 groups. Conclusions and Relevance: Results of this trial showed that acupoint hot compress after vaginal delivery decreased postpartum urinary retention, uterine contraction pain, and depressive symptoms and increased breastfeeding milk volume. Acupoint hot compress may be considered as an adjunctive intervention in postnatal care that meets patient self-care needs. Trial Registration: Chinese Clinical Trial Registry Identifier: ChiCTR2000038417.

Changes in obesity and lifestyle behaviours during the COVID-19 pandemic in Chinese adolescents: A longitudinal analysis from 2019 to 2020.

BACKGROUND: Since December 2019, the coronavirus disease 2019 (COVID-19) has become a global pandemic. Currently, the COVID-19 pandemic is still ongoing. What changes have taken place in the obesity and obesity-related lifestyle behaviours of adolescents during the first year of the COVID-19 pandemic? OBJECTIVE: This study aims at analysing the changes in obesity and lifestyle behaviours of Chinese adolescents before and 1 year after the outbreak of the COVID-19 pandemic, providing evidence for the global strategies to respond to the impact of the COVID-19 pandemic on adolescent obesity. METHODS: Physical examinations and student health and influencing factors questionnaires were conducted among 6047 adolescents aged 11-16 years by health professionals in Shanghai, China, before the COVID-19 pandemic (September-November of 2019) and 1 year after the outbreak of the COVID-19 pandemic (September-November of 2020). Paired χ2 tests, paired t-tests or Wilcoxon signed-rank test was used to evaluate the changes in the obesity prevalence, BMI and lifestyle behaviours from 2019 to 2020. RESULTS: 1 year after the outbreak of the COVID-19 pandemic, the obesity prevalence of Chinese adolescents rose from 14.2% to 15.4% (p < 0.01), mainly because of the increase in boys. And the average BMI increased from 20.3 to 21.2 kg/m2 (p < 0.01). Their lifestyle behaviours have also significantly changed. The mobile screen time increased from 0.25-1.50 h/day to 0.33-2.00 h/day (p < 0.01). The proportion of adolescents who participated in MVPA for ≥60 min/day on all 7 days during the past week dropped from 14.4% to 11.7% (p < 0.01). The generalized estimation equation analysis indicated that adolescents who participated in MVPA for ≥60 min/day on all 7 days had a lower likelihood of having obesity. Boys with computer time ≥2 h/day and girls with mobile screen time ≥2 h/day or TV time ≥2 h/day had a higher likelihood of having obesity. CONCLUSION: This study found that 1 year after the outbreak of the COVID-19 pandemic, the BMI and obesity prevalence of Chinese adolescents increased and obesity-related lifestyle behaviours have also changed.

Mutation of the EPHA2 Tyrosine-Kinase Domain Dysregulates Cell Pattern Formation and Cytoskeletal Gene Expression in the Lens.

Genetic variations in ephrin type-A receptor 2 (EPHA2) have been associated with inherited and age-related forms of cataract in humans. Here, we have characterized the eye lens phenotype and transcript profile of germline Epha2 knock-in mutant mice homozygous for either a missense variant associated with age-related cataract in humans (Epha2-Q722) or a novel insertion-deletion mutation (Epha2-indel722) that were both located within the tyrosine-kinase domain of EPHA2. Confocal imaging of ex vivo lenses from Epha2-indel722 mice on a fluorescent reporter background revealed misalignment of epithelial-to-fiber cell meridional-rows at the lens equator and severe disturbance of Y-suture formation at the lens poles, whereas Epha2-Q722 lenses displayed mild disturbance of posterior sutures. Immunofluorescent labeling showed that EPHA2 was localized to radial columns of hexagonal fiber cell membranes in Epha2-Q722 lenses, whereas Epha2-indel722 lenses displayed disorganized radial cell columns and cytoplasmic retention of EPHA2. Immunoprecipitation/blotting studies indicated that EPHA2 formed strong complexes with Src kinase and was mostly serine phosphorylated in the lens. RNA sequencing analysis revealed differential expression of several cytoskeleton-associated genes in Epha2-mutant and Epha2-null lenses including shared downregulation of Lgsn and Clic5. Collectively, our data suggest that mutations within the tyrosine-kinase domain of EPHA2 result in lens cell patterning defects and dysregulated expression of several cytoskeleton-associated proteins.

Primary Hyperparathyroidism in Pregnancy Followed by Successful Delivery: a Case Report.

BACKGROUND: Primary hyperparathyroidism (PHPT) in pregnancy has a negative impact. Effective treatment should be timely adopted. METHODS: We report a case of a 24-year-old pregnant woman admitted with PHPT, hypercalcemia crisis, hypokalemia, thyroid nodules, hyperthyroidism, and intrauterine single live fetus in the 2nd trimester of pregnancy. Right parathyroidectomy and partial thyroidectomy were timely performed. Postoperative pathology suggested parathyroid adenoma with capsule invasion and thyroid nodules. RESULTS: Postoperative serum PTH and Ca2+ were effectively reduced. Eventually, a healthy fetus was delivered via cesarean at full term. CONCLUSIONS: Parathyroidectomy within reasonable operative timing can improve maternal and fetal prognosis in PHPT during pregnancy, especially with concomitant hypercalcemia crisis.

How are emotional distress and reassurance expressed in medical consultations for people with long-term conditions who were unable to receive curative treatment? A pilot observational study with huntington's disease and prostate cancer.

OBJECTIVE: It is unclear whether how people with long-term conditions express distress, and how clinicians respond, influences perceptions of consultation outcomes. The pilot study examined emotional distress and reassurance in consultations with people whose long-term conditions (at the time of consultations) were treated using active surveillance or symptom management (as no curative treatment was suitable). METHODS: An observational pilot study was conducted involving consultations between people with long-term conditions and their respective clinician. Consultations between three clinicians (two Huntington's Disease; one Prostate Cancer) and 22 people with long-term conditions (11 Huntington's Disease; 11 Prostate Cancer) were audio-recorded. Participants also completed an expanded Consultation and Relational Empathy (CARE) Measure. Two researchers coded sessions using Verona Coding Definitions of Emotional Sequences (VR-CoDES/VR-CoDES-P). Code frequencies were calculated, t tests performed between conditions, and Pearson's correlations performed for associations between CARE responses and clinician utterances. RESULTS: People with long-term conditions expressed emotional distress on average 4.45 times per session, averaging 1.09 Concern and 3.36 Cue utterances. Clinicians responded with more explicit (2.59) and space-providing (3.36), than non-explicit (1.86) and space-reducing (1.09), responses per session. Clinicians expressed spontaneous reassurance on average 5.18 times per session, averaging 3.77 Cognitive and 1.5 Affective reassurance utterances. Huntington's Disease consultations featured significantly more 'Cues', 'Concerns' and 'Overall' 'Emotional Distress', and 'Cognitive' and 'Overall' 'Reassurance'. CONCLUSION: Emotional distress was expressed more using hints than explicit concern utterances. Clinicians predominantly explicitly explored distress rather than providing information/advice and provided advice using spontaneous cognitive reassurance. People with Huntington's Disease expressed more concerns and received more reassurance, indicating different needs between conditions. Future research is required to explore emotional distress and reassurance in a larger sample of participants and long-term condition types, and how the practical implications of these findings may be used to enhance outcomes of consultations. TRIAL REGISTRATION: N/A.

Effects of Midwifery Care on Mode of Delivery, Duration of Labor and Postpartum Hemorrhage of Elderly Parturients.

BACKGROUND: We aimed to explore the influence of midwifery care on the mode of delivery (MOD), duration of labor and postpartum hemorrhage (PPH) of elderly parturients. METHODS: A total of 165 elderly parturients admitted to Tongde Hospital of Zhejiang Province, China from May 2018 to Aug 2019 were allocated into a study group (n=85, midwifery care) and a control group (n=80, conventional nursing care). Visual analogue scale (VAS) was employed to estimate the pain of parturients, and self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were employed to evaluate the anxiety and depression status. The satisfaction of patients on nursing care was scored by a self-made satisfaction questionnaire. RESULTS: The parturients in the study group presented higher spontaneous vaginal delivery (SVD) rate (P < 0.05), and had shortened duration of first and second stages of labor and total duration of labor (P < 0.05). The incidence of both PPH and neonatal asphyxia in the study group was lower than that in the control group (both P < 0.05). The VAS, SAS and SDS scores in the study group were significantly lower than those in the control group (P < 0.05), and the patients' satisfaction in the study group was significantly higher (P < 0.05). CONCLUSION: Midwifery care is effective for the delivery of elderly parturients, which significantly improves VSD rate, shortens duration of labor, reduces incidence of PPH and neonatal asphyxia, as well as relives labor pain, anxiety and depression, and increases satisfaction of parturients.

Factors Associated with Medication Adherence for People Living with Acute HIV Infection in a Tertiary Chinese Hospital in Beijing.

Both the management and caregiving intervention of people living with HIV (PLWH), especially during acute HIV-1 infection, represent a public health issue and a form of social support. This current study analyzed the demographic and clinical factors associated with antiretroviral therapy (ART) adherence of PLWH from positive HIV diagnosis to ART initiation in a tertiary Chinese hospital in Beijing. A total of 200 participants diagnosed with acute HIV-1 infection were enrolled in this study. We collected demographic and clinical data by the use of a self-reported questionnaire. Bivariate and multivariate logistic regressions were used to determine associations between potential variables and outcomes. We found that medication adherence was impacted by years of ART and number of reminders (all P < 0.05). In addition, medication adherence was associated with viral load at 48 weeks (P = 0.035). Future studies are needed to investigate effective interventions that could facilitate ART adherence.

Mutation of the TRPM3 cation channel underlies progressive cataract development and lens calcification associated with pro-fibrotic and immune cell responses.

Transient-receptor-potential cation channel, subfamily M, member 3 (TRPM3) serves as a polymodal calcium sensor in diverse mammalian cell-types. Mutation of the human TRPM3 gene (TRPM3) has been linked with inherited forms of early-onset cataract with or without other eye abnormalities. Here, we have characterized the ocular phenotypes of germline "knock-in" mice that harbor a human cataract-associated isoleucine-to-methionine mutation (p.I65M) in TRPM3 (Trpm3-mutant) compared with germline "knock-out" mice that functionally lack TRPM3 (Trpm3-null). Despite strong expression of Trpm3 in lens epithelial cells, neither heterozygous (Trpm3+/- ) nor homozygous (Trpm3-/- ) Trpm3-null mice developed cataract; however, the latter exhibited a mild impairment of lens growth. In contrast, homozygous Trpm3-M/M mutants developed severe, progressive, anterior pyramid-like cataract with microphthalmia, whereas heterozygous Trpm3-I/M and hemizygous Trpm3-M/- mutants developed anterior pyramidal cataract with delayed onset and progression-consistent with a semi-dominant lens phenotype. Histochemical staining revealed abnormal accumulation of calcium phosphate-like deposits and collagen fibrils in Trpm3-mutant lenses and immunoblotting detected increased αII-spectrin cleavage products consistent with calpain hyper-activation. Immunofluorescent confocal microscopy of Trpm3-M/M mutant lenses revealed fiber cell membrane degeneration that was accompanied by accumulation of alpha-smooth muscle actin positive (α-SMA+ve) myofibroblast-like cells and macrosialin positive (CD68+ve) macrophage-like cells. Collectively, our mouse model data support an ocular disease association for TRPM3 in humans and suggest that (1) Trpm3 deficiency impaired lens growth but not lens transparency and (2) Trpm3 dysfunction resulted in progressive lens degeneration and calcification coupled with pro-fibrotic (α-SMA+ve) and immune (CD68+ve) cell responses.

The Human Takes It All: Humanlike Synthesized Voices Are Perceived as Less Eerie and More Likable. Evidence From a Subjective Ratings Study.

Background: The increasing involvement of social robots in human lives raises the question as to how humans perceive social robots. Little is known about human perception of synthesized voices. Aim: To investigate which synthesized voice parameters predict the speaker's eeriness and voice likability; to determine if individual listener characteristics (e.g., personality, attitude toward robots, age) influence synthesized voice evaluations; and to explore which paralinguistic features subjectively distinguish humans from robots/artificial agents. Methods: 95 adults (62 females) listened to randomly presented audio-clips of three categories: synthesized (Watson, IBM), humanoid (robot Sophia, Hanson Robotics), and human voices (five clips/category). Voices were rated on intelligibility, prosody, trustworthiness, confidence, enthusiasm, pleasantness, human-likeness, likability, and naturalness. Speakers were rated on appeal, credibility, human-likeness, and eeriness. Participants' personality traits, attitudes to robots, and demographics were obtained. Results: The human voice and human speaker characteristics received reliably higher scores on all dimensions except for eeriness. Synthesized voice ratings were positively related to participants' agreeableness and neuroticism. Females rated synthesized voices more positively on most dimensions. Surprisingly, interest in social robots and attitudes toward robots played almost no role in voice evaluation. Contrary to the expectations of an uncanny valley, when the ratings of human-likeness for both the voice and the speaker characteristics were higher, they seemed less eerie to the participants. Moreover, when the speaker's voice was more humanlike, it was more liked by the participants. This latter point was only applicable to one of the synthesized voices. Finally, pleasantness and trustworthiness of the synthesized voice predicted the likability of the speaker's voice. Qualitative content analysis identified intonation, sound, emotion, and imageability/embodiment as diagnostic features. Discussion: Humans clearly prefer human voices, but manipulating diagnostic speech features might increase acceptance of synthesized voices and thereby support human-robot interaction. There is limited evidence that human-likeness of a voice is negatively linked to the perceived eeriness of the speaker.

CLG from Hemp Seed Inhibits LPS-Stimulated Neuroinflammation in BV2 Microglia by Regulating NF-κB and Nrf-2 Pathways.

The healthy benefits of hemp (Cannabis sativa L.) seed have often been attributed to its oils and proteins. Recent studies reveal that hemp seed phenylpropionamides could also show various bioactivities. Continuation of our study on hemp seed provided a phenylpropionamide, coumaroylaminobutanol glucopyranoside (CLG). This work investigated the neuroprotective effect of CLG and its underlying mechanism using lipopolysaccharide-induced BV2 microglia. Our study demonstrated that CLG increased adenosine monophosphate-activated protein kinase (AMPK) expression, suppressed the nuclear factor-kappa B (NF-κB) signaling pathway by inhibiting the phosphorylation of IκBα and NF-κB p65 and decreased proinflammatory cytokine levels in a concentration-dependent manner. Furthermore, CLG reduced the production of cellular reactive oxygen species and stimulated the nuclear factor erythroid 2-related factor 2 (Nrf-2) signaling pathway. Collectively, these results suggested that CLG effectively and simultaneously inhibited inflammatory responses and oxidative stress through the NF-κB and Nrf-2 signaling pathways. AMPK was also involved in the anti-inflammatory effect of CLG. This study provides new insights into the diverse bioactive constituents of hemp seed.

A charged multivesicular body protein (CHMP4B) is required for lens growth and differentiation.

Charged multivesicular body protein 4B (CHMP4B) functions as a core component of the endosome sorting complex required for transport-III (ESCRT-III) machinery that facilitates diverse membrane remodeling and scission processes in eukaryotes. Mutations in the human CHMP4B gene underlie rare, inherited forms of early-onset lens opacities or cataract. Here we have characterized the lens phenotypes of mutant (knock-in) mice harboring a human cataract-associated mutation (p.D129V) in CHMP4B (Chmp4b-mutant) and conditional knockdown mice deficient in lens CHMP4B (Chmp4b-CKD). In situ hybridization localized Chmp4b transcripts to lens epithelial cells and elongating fiber cells at the lens equator. Heterozygous Chmp4b-mutant (D/V) mice were viable and fertile with lenses grossly similar to those of wild-type. However, homozygous Chmp4b-mutant (V/V) mice died by embryonic day 15.5 (E15.5) with grossly abnormal eye and brain histology. Chmp4b-CKD mice displayed variable degrees of lens dysmorphology including lens ablation. Immuno-localization of aquaporin-0 (AQP0) revealed lens fiber cell degeneration in homozygous Chmp4b-mutant (V/V) mouse embryos and in embryonic and postnatal Chmp4b-CKD mice. DNA fragmentation (TUNEL) analysis revealed global cell death in homozygous Chmp4b-mutant (V/V) embryos, whereas, cell death was confined to the lens of Chmp4b-CKD mice. Immuno-localization of the monocyte/macrophage marker macrosialin (CD68) suggested that severe lens degeneration in Chmp4b-CKD mice resulted in an ocular immune cell response. Collectively, these mouse data suggest that (1) heterozygous, germ-line mutations in Chmp4b may not manifest as cataract, (2) homozygous, germ-line mutations in Chmp4b are embryonic lethal, and (3) conditional loss of Chmp4b results in arrest of lens growth and differentiation.

Hemp (Cannabis sativa L.) Seed Phenylpropionamides Composition and Effects on Memory Dysfunction and Biomarkers of Neuroinflammation Induced by Lipopolysaccharide in Mice.

Hempseed has achieved a growing popularity in human nutrition, particularly regarding essential amino acids and fatty acids. The multiple positive attributes of hempseed have led to the further study of its constituents. In this study, hempseed extract containing phenylpropionamides (TPA) was obtained and its chemical profile and content were obtained using high-performance liquid chromatography technology based on previous study. The anti-neuroinflammatory effect of TPA extract was evaluated using a lipopolysaccharide (LPS)-induced mouse model. Fourteen phenylpropionamides (TPA) were identified in the obtained extract with a total content of 233.52 ± 2.50 µg/mg extract. In mice, TPA prevented the learning and spatial memory damage induced by LPS. Increased brain levels of IL-1ß, IL-6, and TNF-α in the LPS-induced mice were reduced by TPA treatment. Furthermore, TPA attenuated LPS-induced hippocampal neuronal damage in mice. This study demonstrates the nutraceutical potential of hempseed from a neuroprotective perspective.

A mixed-methods feasibility study protocol to assess the communication behaviours within the dental health professional-parent-child triad in a general dental practice setting.

BACKGROUND: The promotion of twice yearly application of fluoride varnish (FVA) to the teeth of pre-school children in the dental practice is one component of Scotland's child oral health improvement programme (Childsmile). Nevertheless, evidence shows that application rates of FVA are variable and below optimal levels. The reasons are complex, with many contextual factors influencing activity. However, we propose that one possible reason may be related to the communication challenges when interacting with younger children. Therefore, the primary aim of the study is to assess the feasibility of conducting a video observational study in primary dental care. The secondary aim is to assess the communication behaviours of dental professionals and those of the parents to predict child cooperation when receiving FVA using this video observational study design. METHODS: Approximately 50 eligible pairs of parents and child patients aged between 2 years and 5 years from general dental practices will be recruited to participate in the study. The consecutive mixed-method study will consist of two parts. The first part will be cross-sectional observations of the dental health professional-child-parent communication during dental appointments conducted in the general dental practice setting, using video recording. The second part will be a post-observation, semi-structured interview with parents and dental health professionals respectively. This will be implemented to explore their views on the acceptability and feasibility of being observed using video cameras during treatment provision. DISCUSSION: The mixed-methods study will allow for directly observing the communication behaviours in the clinical setting and uncovering the views of participating dental health professionals and parents. Therefore, the study will enable us to [i] explore new ways to study the nature of triadic interaction of dental health professional-child-parent, [ii] identify dental health professionals' effective communication behaviours that promote child patient and parent's experience of using preventive dental service and [iii] to assess the feasibility of the study through uncovering the views of dental health professionals and parents.

KIR3DL1-Negative CD8 T Cells and KIR3DL1-Negative Natural Killer Cells Contribute to the Advantageous Control of Early Human Immunodeficiency Virus Type 1 Infection in HLA-B Bw4 Homozygous Individuals.

Bw4 homozygosity in human leukocyte antigen class B alleles has been associated with a delayed acquired immunodeficiency syndrome (AIDS) development and better control of human immunodeficiency virus type 1 (HIV-1) viral load (VL) than Bw6 homozygosity. Efficient CD8 T cell and natural killer (NK) cell functions have been described to restrain HIV-1 replication. However, the role of KIR3DL1 expression on these cells was not assessed in Bw4-homozygous participants infected with HIV-1 CRF01_A/E subtype, currently the most prevalent subtype in China. Here, we found that the frequency of KIR3DL1-expressing CD8 T cells of individuals homozygous for Bw6 [1.53% (0-4.56%)] was associated with a higher VL set point (Spearman rs = 0.59, P = 0.019), but this frequency of KIR3DL1+CD8+ T cells [1.37% (0.04-6.14%)] was inversely correlated with CD4 T-cell count in individuals homozygous for Bw4 (rs = -0.59, P = 0.011). Moreover, CD69 and Ki67 were more frequently expressed in KIR3DL1-CD8+ T cells in individuals homozygous for Bw4 than Bw6 (P = 0.046 for CD69; P = 0.044 for Ki67), although these molecules were less frequently expressed in KIR3DL1+CD8+ T cells than in KIR3DL1-CD8+ T cells in both groups (all P < 0.05). KIR3DL1-CD8+ T cells have stronger p24-specific CD8+ T-cell responses secreting IFN-γ and CD107a than KIR3DL1+CD8+ T cells in both groups (all P < 0.05). Thus, KIR3DL1 expression on CD8 T cells were associated with the loss of multiple functions. Interestingly, CD69+NK cells lacking KIR3DL1 expression were inversely correlated with HIV-1 VL set point in Bw4-homozygous individuals (rs = -0.52, P = 0.035). Therefore, KIR3DL1-CD8+ T cells with strong early activation and proliferation may, together with KIR3DL1-CD69+NK cells, play a protective role during acute/early HIV infection in individuals homozygous for Bw4. These findings highlight the superior functions of KIR3DL1-CD8+ T cells and KIR3DL1-CD69+NK cells being a potential factor contributing to delayed disease progression in the early stages of HIV-1 infection.