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.

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.

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.

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.

Epha2 and Efna5 participate in lens cell pattern-formation.

Ephrin type-A receptor 2 (EPHA2) and one of its ligands, ephrin-A5 (EFNA5), have been associated with loss of eye lens transparency, or cataract, - an important cause of visual impairment. Here we show that mice functionally lacking EPHA2 (Epha2-null), EFNA5 (Efna5-null), or both receptor and ligand (Epha2/Efna5-null) consistently develop mostly transparent lenses with an internal refractive disturbance and a grossly disturbed cellular architecture. In situ hybridization localized Epha2 and Efna5 transcripts to lens epithelial cells and nascent fiber cells at the lens equator. In vivo labeling of Epha2-null lenses with a thymidine analog detected a significant decrease in lens epithelial cell proliferation within the germinative zone resulting in impaired early lens growth. Ex vivo imaging of Epha2-null, Efna5-null, and Epha2/Efna5-null lenses labelled in vivo with a membrane-targeted red fluorescent protein revealed misalignment of elongating fiber cells at the lens equator and loss of Y-suture pattern formation near the anterior and posterior poles of the lens. Immuno-fluorescent labeling of lens major intrinsic protein or aquaporin-0 (MIP/AQP0) showed that the precise, radial column patterning of hexagonal fiber cells throughout the cortex region was disrupted in Epha2-null, Efna5-null and Epha2/Efna5-null lenses. Collectively, these data suggest that Epha2 and Efna5 participate in the complex, global patterning of lens fiber cells that is necessary for maximal optical quality.

Multimorbidity and Socioeconomic Deprivation in Primary Care Consultations.

PURPOSE: The influence of multimorbidity on the clinical encounter is poorly understood, especially in areas of high socioeconomic deprivation where burdensome multimorbidity is concentrated. The aim of the current study was to examine the effect of multimorbidity on general practice consultations, in areas of high and low deprivation. METHODS: We conducted secondary analyses of 659 video-recorded routine consultations involving 25 general practitioners (GPs) in deprived areas and 22 in affluent areas of Scotland. Patients rated the GP's empathy using the Consultation and Relational Empathy (CARE) measure immediately after the consultation. Videos were analyzed using the Measure of Patient-Centered Communication. Multilevel, multi-regression analysis identified differences between the groups. RESULTS: In affluent areas, patients with multimorbidity received longer consultations than patients without multimorbidity (mean 12.8 minutes vs 9.3, respectively; P = .015), but this was not so in deprived areas (mean 9.9 minutes vs 10.0 respectively; P = .774). In affluent areas, patients with multimorbidity perceived their GP as more empathic (P = .009) than patients without multimorbidity; this difference was not found in deprived areas (P = .344). Video analysis showed that GPs in affluent areas were more attentive to the disease and illness experience in patients with multimorbidity (P < .031) compared with patients without multimorbidity. This was not the case in deprived areas (P = .727). CONCLUSIONS: In deprived areas, the greater need of patients with multimorbidity is not reflected in the longer consultation length, higher GP patient centeredness, and higher perceived GP empathy found in affluent areas. Action is required to redress this mismatch of need and service provision for patients with multimorbidity if health inequalities are to be narrowed rather than widened by primary care.

Lens ER-stress response during cataract development in Mip-mutant mice.

Major intrinsic protein (MIP) is a functional water-channel (AQP0) that also plays a key role in establishing lens fiber cell architecture. Genetic variants of MIP have been associated with inherited and age-related forms of cataract; however, the underlying pathogenic mechanisms are unclear. Here we have used lens transcriptome profiling by microarray-hybridization and qPCR to identify pathogenic changes during cataract development in Mip-mutant (Lop/+) mice. In postnatal Lop/+ lenses (P7) 99 genes were up-regulated and 75 were down-regulated (>2-fold, p=<0.05) when compared with wild-type. A pathway analysis of up-regulated genes in the Lop/+ lens (P7) was consistent with endoplasmic reticulum (ER)-stress and activation of the unfolded protein response (UPR). The most up-regulated UPR genes (>4-fold) in the Lop/+ lens included Chac1>Ddit3>Atf3>Trib3>Xbp1 and the most down-regulated genes (>5-fold) included two anti-oxidant genes, Hspb1 and Hmox1. Lop/+ lenses were further characterized by abundant TUNEL-positive nuclei within central degenerating fiber cells, glutathione depletion, free-radical overproduction, and calpain hyper-activation. These data suggest that Lop/+ lenses undergo proteotoxic ER-stress induced cell-death resulting from prolonged activation of the Eif2ak3/Perk-Atf4-Ddit3-Chac1 branch of the UPR coupled with severe oxidative-stress.

Lens transcriptome profile during cataract development in Mip-null mice.

Major intrinsic protein or aquaporin-0 (MIP/AQP0) functions as a water channel and a cell-junction molecule in the vertebrate eye lens. Loss of MIP function in the lens leads to degraded optical quality and cataract formation by pathogenic mechanisms that are unclear. Here we have used microarray-hybridization analysis to detect lens transcriptome changes during cataract formation in mice that are functionally null for MIP (Mip-/-). In newborn Mip-/- lenses (P1) 11 genes were up-regulated and 18 were down-regulated (>2-fold, p=<0.05) and a similar number of genes was differentially regulated at P7. The most up-regulated genes (>6-fold) in the Mip-/- lens at P1 included those coding for a mitochondrial translocase (Timmdc1), a matrix metallopeptidase (Mmp2), a Rho GTPase-interacting protein (Ubxn11) and a transcription factor (Twist2). Apart from Mip, the most down-regulated genes (>4-fold) in the Mip-/- lens at P1 included those coding for a proteasome sub-unit (Psmd8), a ribonuclease (Pop4), and a heat-shock protein (Hspb1). Lens fiber cell degeneration in the Mip-/- lens was associated with increased numbers of TUNEL-positive cell nuclei and dramatically elevated levels of calpain-mediated proteolysis of αII-spectrin. However red-ox status, measured by glutathione and free-radical levels, was similar to that of wild-type. These data suggest that while relatively few genes (∼1.5% of the transcriptome) were differentially regulated >2-fold in the Mip-/- lens, calpain hyper-activation acts as a terminal pathogenic event during lens fiber cell death and cataract formation.

Novel Galactosylated Poly(ethylene glycol)-Cholesterol for Liposomes as a Drug Carrier for Hepatocyte-Targeting.

In this study, three types of galactosylated cholesterol (i.e., gal-PEG194-chol, gal-PEG1000-chol and gal-PEG2000-chol) were synthesized with one terminal of polyethylene glycol of various chain lengths conjugated to the galactoside moiety, and the other terminal conjugated to the cholesterol. The galactose-modified liposomes were prepared by thin film-hydration method and doxorubicin (DOX) was loaded to the liposomes by using a ammonium sulfate gradient procedure. The liposomal formulations with galactosylated cholesterol were characterized. Flow cytometry and laser confocal scanning microscopy analyses showed that the galactose-modified liposomes facilitated the intracellular uptake of liposomes into HepG2 via asialoglycoprotein receptor (ASGP-R) mediated endocytosis. Cytotoxicity assay showed that the cell proliferation inhibition effect of galactose-modified liposomes was higher than that of the unmodified liposomes. Additionally, the study on frozen section of liver showed that the galactose-modified liposomes enhanced the intracellular uptake of liposomes into hepatocytes. Taken together, these results suggested that liposomes containing such galactosylated cholesterol (i.e., gal-PEG-chol), had a great potential as drug delivery carriers for hepatocyte-selective targeting.

How head and neck consultants manage patients' emotional distress during cancer follow-up consultations: a multilevel study.

Head and neck cancer (HNC) patients suffer substantial emotional problems. This study aimed to explore how utterance-level variables (source, type and timing of emotional cues) and patient-level variables (e.g. age, gender and emotional well-being) relate to consultants' responses (i.e. reducing or providing space) to patient expressions of emotional distress. Forty-three HNC outpatient follow-up consultations were audio recorded and coded, for patients' expressions of emotional distress and consultants' responses, using the Verona Coding Definitions of Emotional Sequence. Multilevel logistic regression modelled the probability of the occurrence of consultant-reduced space response as a function of patient distress cue expression, controlling for consultation and patient-related variables. An average of 3.5 cues/concerns (range 1-20) was identified per consultation where 84 out of 152 total cues/concerns were responded by reducing space. Cue type did not impact on response; likewise for the quality of patient emotional well-being. However, consultants were more likely to reduce space to cues elicited by patients, as opposed to those initiated by themselves. This reduced space response was more pronounced as the consultation continued. However, about 6 min into the consultation, this effect (i.e. tendency to block patients) started to weaken. Head and neck consultants' responses to negative emotions depended on source and timing of patient emotional expressions. The findings are useful for training programme development to encourage consultants to be more flexible and open in the early stages of the consultation.

Reassurance and distress behavior in preschool children undergoing dental preventive care procedures in a community setting: a multilevel observational study.

BACKGROUND: The effect of reassurance in managing distress among children who receive procedures of a less aversive nature has not been fully investigated. PURPOSE: This study aimed to investigate the relationship between reassurance by dental staff and distress behavior of preschool children receiving preventive procedures in a community setting. METHODS: Nurse-child interactions (n = 270) during fluoride varnish application were video recorded and coded. Multilevel logistic regression modeled the probability of the occurrence of child distress behavior as a function of reassurance provision, controlling for child-level and nurse-level variables. RESULTS: Child distress behavior was positively related to nurse verbal reassurance but negatively linked to the time that this reassurance occurred. Both child initial anxiety and nurse nonprocedural training increased the probability of observable distress behavior. CONCLUSIONS: The use of verbal reassurance to promote reception of mild invasive procedures was counterindicated, especially when offered early in the intervention ( ClinicalTrials.gov number: NCT00881790).

Laminin ß2 gene missense mutation produces endoplasmic reticulum stress in podocytes.

Mutations in the laminin ß2 gene (LAMB2) cause Pierson syndrome, a severe congenital nephrotic syndrome with ocular and neurologic defects. LAMB2 is a component of the laminin-521 (α5ß2γ1) trimer, an important constituent of the glomerular basement membrane (GBM). The C321R-LAMB2 missense mutation leads to congenital nephrotic syndrome but only mild extrarenal symptoms; the mechanisms underlying the development of proteinuria with this mutation are unclear. We generated three transgenic mouse lines, in which rat C321R-LAMB2 replaced mouse LAMB2 in the GBM. During the first postnatal month, expression of C321R-LAMB2 attenuated the severe proteinuria exhibited by Lamb2(-/-) mice in a dose-dependent fashion; proteinuria eventually increased, however, leading to renal failure. The C321R mutation caused defective secretion of laminin-521 from podocytes to the GBM accompanied by podocyte endoplasmic reticulum (ER) stress, likely resulting from protein misfolding. Moreover, ER stress preceded the onset of significant proteinuria and was manifested by induction of the ER-initiated apoptotic signal C/EBP homologous protein (CHOP), ER distention, and podocyte injury. Treatment of cells expressing C321R-LAMB2 with the chemical chaperone taurodeoxycholic acid (TUDCA), which can facilitate protein folding and trafficking, greatly increased the secretion of the mutant LAMB2. Taken together, these results suggest that the mild variant of Pierson syndrome caused by the C321R-LAMB2 mutation may be a prototypical ER storage disease, which may benefit from treatment approaches that target the handling of misfolded proteins.

Prediction of nursery school-aged children who refuse fluoride varnish administration in a community setting: a Childsmile investigation.

BACKGROUND: Young children of pre-school age may find a minimal intervention (fluoride varnish application) difficult to tolerate. AIM: To determine the significant predictors for refusing a fluoride varnish application from child, parental and nurse behaviour factors. DESIGN: Data included videos from 238 children (52% female, aged 3-5 years) receiving a fluoride varnish application in a Scottish nursery school setting. The St Andrews Behavioural Interaction Scheme (SABICS) was used for video coding and retrieved child refusal status, initial anxious behaviour, and nurse behaviour. A parental survey collected parent's dental anxiety [Modified Dental Anxiety Scale (MDAS)] and the child's home behaviour [Strengths and Difficulties Questionnaire (SDQ)]. Child demographics, dental status, and previous varnish application experience were recorded. Multivariate binary logistic regression was applied to predict child refusal of the varnish application. RESULTS: The response rate was 79%. Twelve children refused. The significant predictors of varnish refusal included initial anxious child behaviour (ß = 5.14, P = 0.001), no previous varnish application (ß = -3.89, P = 0.04), and no nurse praise (ß = -1.06, P = 0.02). Information giving (P = 0.06) and reassurance (P = 0.08) were borderline significant. CONCLUSION: Initial anxiety behaviour, previous varnish experience, and not using praise by the nursing staff predicted fluoride varnish application refusal.

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.

Changes in group II phospholipase A2 gene expression in rat heart during sepsis.

BACKGROUND: This study was undertaken to investigate alterations of group II phospholipase A2 (PLA2) gene expression and its underlying mechanism in rat heart during different phases of sepsis. MATERIALS AND METHODS: Sepsis was induced by cecal ligation and puncture (CLP). Experiments were divided into three groups, control, early sepsis, and late sepsis. Early and late sepsis refers to those animals sacrificed at 9 and 18 h, respectively, after CLP. PLA2 enzyme activity, group II PLA2 protein level, messenger RNA (mRNA) abundance, transcription rate, and half-life were measured. RESULTS: PLA2 activity was decreased by 29% during early sepsis but it was increased by 49% during late sepsis. Group II PLA2 protein level was decreased by 27% during early sepsis but it was increased by 35.3% during late sepsis. Group II PLA2 mRNA was decreased by 21% during early sepsis but it was increased by 141% during late sepsis. The transcription rate of group II PLA2 mRNA was reduced by 25% during early sepsis but it was elevated by 67% during late sepsis. The half-life of group II PLA2 mRNA remained unaltered during early and late phases of sepsis. CONCLUSIONS: These results demonstrate that PLA2 activity, group II PLA2 protein level, the mRNA abundance, and transcription rate were concurrently underexpressed during early sepsis, while they were overexpressed during late sepsis, with no change in the degradation of gene transcript. These data indicate that the biphasic changes in group II PLA2 gene expression are regulated transcriptionally during sepsis.

Prevalence of hypertension in overweight and obese children from a large school-based population in Shanghai, China.

BACKGROUND: The ongoing rise in the prevalence of hypertension in children and adolescents is considered to be accompanied with the epidemic of childhood overweight and obesity. In this study, we established a large scale cross-sectional study in Shanghai, China, which presented a new evidence for the correlation of hypertension prevalence with overweight and obesity stages in Chinese children and adolescents. METHODS: A school-based cross-sectional study was conducted during February to December 2009 in Shanghai, China, including total 78,114 children and adolescents. Body weight, height, waist circumference (WC) and blood pressure (BP) were measured. Overweight and obesity were defined according to sex- and age- specific Chinese reference data. RESULTS: Both SBP and DBP were very significantly increased in overweight (OW) and obese (OB) groups. With age and sex controlled, BMI and WC were independently positively correlated with SBP and DBP. The prevalence of high SBP, DBP and hypertension were markedly higher among OW and OB children than normal weight (NW) group. Odds ratios (ORs) for high SBP, high DBP and high BP were significantly greater in OW and OB children than NW group, and showed a trend increase correlating with obesity stages (all P <0.0001). According to the increasing OR with different combination of obese status of BMI and WC, WC has a stronger influence on hypertension. The combination of BMI and WC obese shows substantially higher ORs compared with those for either BMI or WC obese alone. CONCLUSIONS: In this study on a large school-based population in Shanghai, China, BMI and WC are positively correlated with SBP and DBP. Being overweight or obese greatly increased the risk of hypertension in Chinese children and adolescents, in which WC is considered as a more sensitive indicator than BMI.

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.

"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.

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.

Antisense inhibition of secretory and cytosolic phospholipase A2 reduces the mortality in rats with sepsis*.

OBJECTIVE: Phospholipase A(2) has been implicated to play a pivotal role in the pathogenesis of sepsis syndrome. The two major forms of phospholipase A(2) isoenzymes, secretory phospholipase A(2) and cytosolic phospholipase A(2), are overexpressed during sepsis. The objective of this study was to test the hypothesis that inhibition of the overexpressed secretory phospholipase A(2) and cytosolic phospholipase A(2) during sepsis benefits the disease's eventual outcome. DESIGN: Short-chain antisense oligonucleotide molecules were designed with the aid of computer software programs, and their in vitro efficacies were assessed in cell culture systems based on inhibition of target protein expression. The in vivo efficacies were determined in intact sepsis rats using 35-day survival rate as a primary efficacy end point. SETTING: Animal research laboratory at a university. SUBJECTS: Male Sprague-Dawley rats (180-200 g). INTERVENTIONS: Sepsis was induced by cecal ligation and puncture. Antibiotics were administered subcutaneously once daily at 12 mg/kg, for 20 days. Oligonucleotides (antisense or mismatch) were administered intravenously once daily at 2 mg/kg to 0.8 mg/kg in a decreasing order, for 20 days. MEASUREMENTS AND MAIN RESULTS: In cell culture systems, 21 of the 105 antisense constructs were found to be efficacious in inhibiting secretory phospholipase A(2) IIa and cytosolic phospholipase A(2) IVa protein expression. In sepsis rats, antisense oligonucleotides were capable of reducing their target protein expression by 18%-61% in major organs such as liver, heart, and kidney. In animal experiments, sepsis without any treatment (Group 1) had a median survival time of 2 days and a zero (0) percent survival rate at day 14. Sepsis with antibiotic treatment (Group 2) had a median survival time of 6 days and a 35-day survival rate of 28%. Sepsis with cotreatment of antibiotics and antisense oligonucleotides (one against secretory phospholipase A2 IIa and the other against cytosolic phospholipase A(2) IVa) (Group 4) increased the median survival time from 6 to 35 days and the 35-day survival rate from 28% to 58.8% as compared with antibiotics alone (Group 4 vs. Group 2; p <.05). Sepsis with cotreatment of antibiotics and mismatch oligonucleotides (Group 3) did not affect the median survival time and the 35-day survival rate as compared to antibiotics alone (Group 3 vs. Group 2; p >.05). CONCLUSIONS: The results demonstrate that antisense strategy against secretory phospholipase A(2) IIa and cytosolic phospholipase A(2) IVa can inhibit their target protein expression in major organs and greatly improve the clinical outcome, i.e., an absolute reduction in 35-day mortality of 30.8%, in rats with sepsis. Our studies, thus, provide an improved method for the treatment of sepsis by targeting multiple forms of phospholipase A(2) isoenzymes with DNA antisense oligomers.