OBJECTIVES: The mangrove cricket, Apteronemobius asahinai, shows endogenous activity rhythms that synchronize with the tidal cycle (i.e., a free-running rhythm with a period of ~ 12.4 h [the circatidal rhythm]). Little is known about the molecular mechanisms underlying the circatidal rhythm. We present the draft genome of the mangrove cricket to facilitate future molecular studies of the molecular mechanisms behind this rhythm. DATA DESCRIPTION: The draft genome contains 151,060 scaffolds with a total length of 1.68 Gb (N50: 27 kb) and 92% BUSCO completeness. We obtained 28,831 predicted genes, of which 19,896 (69%) were successfully annotated using at least one of two databases (UniProtKB/SwissProt database and Pfam database).

OBJECTIVE: Lipofection-mediated introduction of the CRISPR/Cas9 system in porcine zygotes provides a simple method for gene editing, without requiring micromanipulation. However, the gene editing efficiency is inadequate. The aim of this study was to improve the lipofection-mediated gene editing efficiency by optimizing the timing and duration of lipofection. RESULTS: Zona pellucida (ZP)-free zygotes collected at 5, 10, and 15 h from the start of in vitro fertilization (IVF) were incubated with lipofection reagent, guide RNA (gRNA) targeting GGTA1, and Cas9 for 5 h. Lipofection of zygotes collected at 10 and 15 h from the start of IVF yielded mutant blastocysts. Next, ZP-free zygotes collected at 10 h from the start of IVF were incubated with lipofection reagent, gRNA, and Cas9 for 2.5, 5, 10, or 20 h. The blastocyst formation rate of zygotes treated for 20 h was significantly lower (p < 0.05) than those of the other groups, and no mutant blastocysts were obtained. Moreover, the mutation rates of the resulting blastocysts decreased as the incubation time increased. In conclusion, a lipofection-mediated gene editing system using the CRISPR/Cas9 system in ZP-zygotes is feasible; however, further improvements in the gene editing efficiency are required.

OBJECTIVE: Due to different concerns in using appropriate mosquito blood feeding techniques, this work focused on evaluating the effectiveness of two artificial mosquito feeding systems (Rutledge and Hemotek) and three different membranes (Parafilm, mouse and chicken skins). Female mosquitoes from an An. coluzzii strain aged between 2 and 5 days were exposed to blood with the two systems at time intervals (5, 10, 15, 20, 25 and 30 min) with blood used on the day of collection, the next day and 2 days after. RESULTS: Our results showed that the Hemotek system gave better blood feeding rates than the Rutledge system. Among the three membranes, the blood feeding rates with chicken and mouse skins were higher than those provided by the Parafilm membrane. Likewise, blood stored 1 day after collection gave higher levels than blood used on the day of collection and 2 days after. Regardless of the system, the lowest blood feeding rates were observed at 5 min compared to the other exposure times.

OBJECTIVE: Charcot foot is a rare complication to neuropathy and can cause severe foot deformities and ulcerations, which often require prolonged antibiotical treatment. The objective of this retrospective study was to investigate whether this treatment is associated to impaired renal function. RESULTS: In total, 163 patients were included, of whom 105 (64%) had received β-lactam antibiotics for a mean total duration of 13.0 months. There was a significant increase in the urine albumin/creatinine ratio in the group that received antibiotics (p = 0.017), and the use of antibiotics was associated to a subsequent diagnosis of nephropathy (p = 0.01). Patients treated with antibiotics had a 21.9% risk of developing subsequent nephropathy versus 5.2% for patients not treated with antibiotics. We suggest increased awareness on signs of nephropathy in patients with severe Charcot foot.

OBJECTIVE: The objective of this study is to assess gender representation in food and beverage print advertisements. RESULTS: The study follows a quantitative descriptive approach. Using a content analysis technique, we assessed the gender representation in 200 food and beverage print advertisements found in corner stores located in four areas around schools in Lima, Peru, and Guatemala City, Guatemala (100 advertisements per country). A total of 36% of the print advertisements exhibited a male main character for the case of Guatemala, while in Peru 14% of the print advertisements presented a male main character. Furthermore, in Guatemala, 22% of the main characters were male animated characters. Moreover, 27% of the print advertisements in Guatemala and 17%, in Peru were visually male-oriented. Overall, male characters appeared alongside sports references and in varied settings, whereas female characters were usually holding or consuming the product. In conclusion, although the majority of variables used to assess the representation of gender in food and beverage print advertisements were gender-neutral, those showing gender representation were mostly male-oriented. Despite its limited findings, the study provides evidence for the formulation of public policies and educational content aimed to protect children's and adolescents' health from the effects of food marketing.

OBJECTIVE: A community-based cross-sectional study was done to assess Plasmodium falciparum exposure in areas with different malaria endemicity in north-eastern Tanzania using serological markers; PfAMA-1 and PfMSP-119. RESULTS: Bondo had a higher seroprevalence 36.6% (188) for PfAMA-1 as compared to Hai 13.8% (33), χ2 = 34.66, p < 0.01. Likewise, Bondo had a higher seroprevalence 201(36.6%) for PfMSP-1 as compared to Hai 41 (17.2%), χ2 = 29.62, p < 0.01. Anti-PfAMA-1 titters were higher in malaria positive individuals (n = 47) than in malaria negative individuals (n = 741) (p = 0.07). Anti-PfMSP-1 antibody concentrations were significantly higher in malaria-positive individuals (n = 47) than in malaria-negative individuals (n = 741) (p = 0.003). Antibody response against PfAMA-1 was significantly different between the three age groups; < 5 years, 5 to 15 years and > 15 years in both sites of Bondo and Hai. Likewise, antibody response against PfMSP-119 was significantly different between the three age groups in the two sites (p < 0.001). We also found significant differences in the anti-PfAMA-1and anti-PfMSP-119 antibody concentrations among the three age groups in the two sites (p = 0.004 and 0.005) respectively. Immunological indicators of P. falciparum exposure have proven to be useful in explaining long-term changes in the transmission dynamics, especially in low transmission settings.

OBJECTIVE: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the novel coronavirus causing severe respiratory illness (COVID-19). This virus was initially identified in Wuhan city, a populated area of the Hubei province in China, and still remains one of the major global health challenges. RNA interference (RNAi) is a mechanism of post-transcriptional gene silencing that plays a crucial role in innate viral defense mechanisms by inhibiting the virus replication as well as expression of various viral proteins. Dicer, Drosha, Ago2, and DGCR8 are essential components of the RNAi system, which is supposed to be dysregulated in COVID-19 patients. This study aimed to assess the expression level of the mentioned mRNAs in COVID-19patients compared to healthy individuals. RESULTS: Our findings demonstrated that the expression of Dicer, Drosha, and Ago2 was statistically altered in COVID-19 patients compared to healthy subjects. Ultimately, the RNA interference mechanism as a crucial antiviral defense system was suggested to be dysregulated in COVID-19 patients.

OBJECTIVE: The clone devaluation is a phenomenon reported by the latest paper in which eeriness is evoked when people observe individuals with the same face (clone faces) compared to those with different faces. There are two possibilities that explain the clone devaluation effect. One is that the same facial features that clone faces have (duplication of facial features) induce the clone devaluation effect. The other possibility is that duplication of identities between people with clone faces is important for the clone devaluation effect. We thus conducted an experiment to investigate whether the duplication of identities or of facial features induces the clone devaluation effect. RESULTS: Participants evaluated eeriness of scrambled clone faces and scrambled different faces using the paired comparison method. There was only a slight difference in subjective eeriness between scrambled clone faces and scrambled different faces. Therefore, this study suggests that the duplication of local facial features does not play a key role in inducing the clone devaluation effect.

OBJECTIVE: Athletic performances are dynamic movements that are physically challenging and often predict individual success in ecological contexts. They stem from a complex integration of multiple phenotypic traits-e.g., morphological, physiological and behavioural-that dictate animal survival and individual fitness. However, directly quantifying athletic performances can be particularly challenging in cryptic, slow-moving species or not very reactive in attitude. Here we present and describe a rapid, simple, and low-cost method to measure athletic performance in post-metamorphic individuals of the fire salamander Salamandra salamandra. While extremely reactive during the larval stage, adult salamanders are, in fact, cryptic and relatively slow-moving. RESULTS: Forcing terrestrial juveniles to swim under standard, albeit ecologically plausible, laboratory conditions, and using an automatic point-mass tracking tool, we were able to measure maximal and average performance indicators of post-metamorphic individuals. This method avoids inter-individual variation in motivation, as it forces individuals to perform at their best. Moreover, with this method, measures of athletic performance will be directly comparable between larval and terrestrial stages, allowing to study the contribution of carryover effects to the wide range of processes implicated in the eco-evo-devo of athletic performance in salamanders.

OBJECTIVE: Pebrine, caused by the microsporidium Nosema bombycis, is one of the severe diseases in Thai polyvoltine strains of the silkworm Bombyx mori. Studies showing the presence of Lactobacillus species in the silkworm gut, where the Nosema parasites enter, suggests that these bacteria may have a protective effect. The aim of this study was to investigate the effect of supplementation of Lactobacillus casei on the survival ratio of silkworm larvae challenged with N. bombycis. RESULTS: A group of silkworm larvae of the commercial Thai polyvoltine hybrid strain DokBua was supplemented with L. casei on the second day of the 2nd, 3rd, 4th, and 5th instar. When a control group of silkworm larvae were challenged with N. bombycis on the second day of the 4th instar, the survival rate was 68%, but it was 91% for larvae supplemented with L. casei. For those larvae that survived the treatments until pupation, we determined the growth characters larval weight, cocooning ratio, and pupation ratio, and the economic characters cocoon weight and cocoon shell weight. When infected with N. Bombycis, growth characters were significantly higher in larvae also receiving L. casei.

OBJECTIVE: Microbial analysis in milk preserved using heat-assisted Pulsed Electric Field (PEF) need to be assessed. In this study we analyze the microbial quality and virulence-associated genes in milk samples preserved using heat-assisted PEF from several producers in Indonesia. RESULTS: Milk samples were collected consisting of raw milk, milks taken after the heating, PEF, mixing, cooling, and packaging. Microbiological and Polymerase Chain Reaction (PCR) detection for virulence genes were performed. Heat-assisted PEF treatment gave 2.7-7.47 log reduction for TPC; 1.6-2.56 log reduction for MPN number; 3.13-6.48 log reduction for S. aureus; and for B. cereus there was an increase of 0.76 log and a reduction of 0.46 log. While milk samples from thermal pasteurization gave log reduction numbers of TPC, MPN, and S. aureus respectively 5.28; 2.56; and 4.73, for B. cereus was increasing 2.4 log. Producer C performed the best results with significant reduction compared with others (p < 0.005). There were no colonies of L. monocytogenes found in all of the samples. PCR results showed that milk samples possessed virulence genes 17.5% (10/57) of invA genes, 54.4% (31/57) of nheA genes, 68.4% (39/57) of cytK genes, 38.6% (22/57) of nuc genes, 63.2% (36/57) of ileS genes, while hly and actA genes were not detected.

OBJECTIVE: To develop a collision engine (haptic force feedback simulator) compatible with a 5-degrees-of-freedom (DOF) haptic wand. This has broad applications such as telerobotic ultrasound systems. Integrating force feedback into systems is critical to optimize remote scanning. A collision engine compatible with a 5-DOF haptic wand was developed based on the Gilbert-Johnson-Keerthi algorithm. The collision engine calculated force during collision between the wand and a virtual object based on code developed using MATLAB. A proportional force was subsequently returned to a user via the haptic wand, thereby simulating the collision force for the user. Three experiments were conducted to assess the accuracy of the collision engine on curved and flat surfaces. RESULTS: The average errors in calculation of distances between the wand and virtual object were 2.1 cm, 3.4 cm, and 4.2 cm for the model of the human hand, cylinder, and cuboid, respectively. The collision engine accurately simulated forces on a flat surface, though was less accurate on curved surfaces. Future work will incorporate haptic force feedback into a telerobotic ultrasound system. The haptic force simulator presented here may also be used in the development of ultrasound simulators for training and education.

OBJECTIVE: In previous studies using Illumina Infinium methylation arrays, we have identified DNA methylation marks associated with cancer predisposition and progression. In the present study, we have sought to find appropriate technology to both technically validate our data and expand our understanding of DNA methylation in these genomic regions. Here, we aimed to assess the repeatability of methylation measures made using QIAseq targeted methyl panel and to compare them with those obtained from the Illumina HumanMethylation450 (HM450K) assay. We included in the analysis high molecular weight DNA extracted from whole blood (WB) and DNA extracted from formalin-fixed paraffin-embedded tissues (FFPE). RESULTS: The repeatability of QIAseq-methylation measures was assessed at 40 CpGs, using the Intraclass Correlation Coefficient (ICC). The mean ICCs and 95% confidence intervals (CI) were 0.72 (0.62-0.81), 0.59 (0.47-0.71) and 0.80 (0.73-0.88) for WB, FFPE and both sample types combined, respectively. For technical replicates measured using QIAseq and HM450K, the mean ICCs (95% CI) were 0.53 (0.39-0.68), 0.43 (0.31-0.56) and 0.70 (0.59-0.80), respectively. Bland-Altman plots indicated good agreement between QIAseq and HM450K measurements. These results demonstrate that the QIAseq targeted methyl panel produces reliable and reproducible methylation measurements across the 40 CpGs that were examined.

OBJECTIVES: Financial protection mechanisms are in place to overcome the costs of a few diseases in India. Our objective was to estimate the costs incurred due to Oral Potentially Malignant Disorders (OPMD) and to determine predictors of such costs. RESULTS: We found that the median (Interquartile range IQR) total costs of OPMD was Indian Rupees (INR) 500 (350-750), direct medical costs was INR 0 (0-50), direct non-medical costs was INR 150 (40-200) and indirect costs was INR 350 (250-500). The travel cost to attend the health facilities [INR 100 (40-150)] and the patient's loss of wages [INR 200 (150-400)] mainly accounted for the direct non-medical and indirect costs respectively. The median expenditure on smokeless and smoking forms of tobacco was INR 6000 (5400-7200) and INR 2400 (1800-3600) respectively. On multiple linear regression analysis, rural residence, belonging to below poverty line family, being a sole earner in the family, number of months since diagnosis and first visit at a private provider were found to be the significant predictors of total costs of OPMD. Financial protection mechanisms are needed for covering the direct non-medical and indirect costs. Early management of OPMD might mitigate the costs of OPMD.

OBJECTIVE: Near visual impairment (VI) is a common disability in an aging population. Near vision is crucial in activity of daily living including reading, smartphone and computer use and meal preparation. This study was conducted to determine the association between near visual acuity (VA) and contrast sensitivity (CS) with activity of daily living (ADL) among visually impaired older adults. RESULTS: A total of 208 participants aged  ≥  60 were recruited from the population-based longitudinal study on neuroprotective model for healthy longevity. Habitual near VA and CS were measured using Lighthouse near VA chart and Pelli-Robson CS chart, respectively. Lawton instrumental activities of daily living (IADL) was used to assess ADL. There are 41.8% participants with near visual impairment and 28.7% among them had IADL disability. Independent t test showed significant lower mean IADL score among visually impaired participants [t(206)  =  2.03, p  =  0.04]. IADL score significantly correlated with near VA (r  =   - 0.21, p  =  0.05) but not with CS (r  =   - 0.14, p = 0.21). Near VA (B  =   - 0.44, p  =  0.03) and age (B  =   - 0.07, p  =  0.01) significantly predicted IADL. The findings show poorer VA renders higher IADL disability, which may necessitate interventions to improve ADL among visually impaired older adults.

OBJECTIVE: Although perceived neighbourhood environment is considered a predictor of leisure-time physical activity (LTPA), evidence for this is limited in South Asia. Thus, the aim was to determine the association between neighbourhood social and built environment features in carrying out LTPA among adults in Colombo District, Sri Lanka. A cross-sectional study among 1320 adults was carried out using validated questionnaires for physical activity (PA) and built environment data collection. Multiple logistic regression analysis was conducted to assess the associations between environment characteristics and LTPA after adjusting for gender, age, employment status, income level and sector of residence. RESULTS: A total of 21.7% of adults participated in some LTPA. The commonest type of LTPA was walking; carried out by 14.5%. Moderate and vigorous activity at leisure was carried out by 10.3% and 3.9% respectively. Perceived social acceptance for PA was positively associated with LTPA. Out of the built environment characteristics perceived infrastructure for walking, and recreational facilities for PA were negatively associated with LTPA. Self-efficacy emerged as an important positive correlate of LTPA. The participants were positively influenced by the self-efficacy and perceived social environment which should be addressed when promoting LTPA.

OBJECTIVE: To identify sociodemographic factors and health conditions associated with self-rated wellbeing for Aboriginal and Torres Strait Islander adults. Participants were recruited via investigator networks and an online panel provider with an established nationwide panel of Aboriginal and Torres Strait Islander adults. Those interested were invited to complete a survey that included an assessment of wellbeing using a visual analogue scale. Data was collected from October-November 2019 and August-September 2020. Exploratory analyses were conducted to ascertain factors associated with self-rated wellbeing for Aboriginal and Torres Strait Islander adults. RESULTS: Having more than enough money to last until next pay day, full-time employment, completion of grade 12, having a partner, and living with others were significantly associated with higher wellbeing among Aboriginal and Torres Strait Islander adults. A self-reported history of depression, anxiety, other mental health conditions, heart disease, or disability were associated with lower self-rated wellbeing scores. Our findings indicate a need for further investigation among these socioeconomic and patient groups to identify how to improve and support the wellbeing of Aboriginal and Torres Strait Islander adults.

Author information: (1)Institute of Health Informatics, University College London, London, UK. (2)Institute of Health Informatics, University College London, London, UK. alvina.lai@ucl.ac.uk.

OBJECTIVES: We investigated the existence and uniqueness of a fixed point for the mapping satisfying generalized rational type contraction conditions in metric space endowed with partial order. Suitable examples are presented to justify the results obtained. RESULT: Some new fixed point results have been obtained for a mapping fulfilling generalized contractions. The uniqueness of the fixed point is also the part of the study based on an ordered relation. One example is given for a result which is not valid in the usual metric space.