Escherichia coli is one of the most favorable hosts for this purpose. Although a number of strategies for optimizing protein production have been developed, the effect of gene overexpression on host cell growth has been much less studied. Here, we performed high-throughput tests on the E. coli a complete set of E. coli K-12 ORF archive (ASKA) collection to quantify the effects of overexpressing individual E. coli genes on its growth. The results indicated that overexpressing membrane-associated proteins or proteins with high abundances of branched-chain amino acids tended to impair cell growth, the latter of which could be remedied by amino acid supplementation. Through this study, we expect to provide an index for a fast pre-study estimate of host cell growth in order to choose proper rescuing approaches when working with different proteins.Recombinant protein production plays an essential role in both biological studies and pharmaceutical production.  After the whole-genome sequences of thousands of organisms have been well documented, overexpressing genes to get highly pure proteins for further characterization and engineering becomes an indispensable part of biochemistry, molecular biology, cell biology, and synthetic biology. Moreover, among the 239 US-FDA (Food and Drug Administration) approved therapeutic peptides and proteins, as well as their 380 drug variants, the majority are manufactured by recombinant protein production . Escherichia coli is one of the most widely-used hosts to express recombinant proteins due to a number of advantages. First, it grows quickly, with a doubling time of about 20 min in rich growth media [13 cells could be obtained from 1 L of liquid Lysogeny broth (LB) medium [E. coli such as the LB medium and the Terrific Broth (TB) medium. Fourth, the genetics of E. coli is well known, and it is convenient to remove certain genes from the genome for different purposes [E. coli by plasmid transformation. Last but not least, a large number of vectors, fusion tags, and mutant strains have been developed for optimal expression of target proteins in E. coli. Several review articles have been published recently to cover these topics [In both basic research and drug production, e topics ,5.A commonly encountered problem for recombinant protein production is impeded cell growth or reduced biomass accumulation. There are two major reasons for this phenomenon. The first is the general metabolic burden, which could be explained as the competition between biomass accumulation and recombinant protein production for metabolic materials such as cellular energy, ATP, and substrates, amino acids, . This coE. coli K-12 ORF archive (ASKA) collection, which is a complete set of E. coli strains for overexpressing individual E. coli K-12 genes [E. coli genes on its own cell growth and combined the results with bioinformatical analyses to identify shared features of proteins, which could hamper cell growth. For this purpose, we utilized the a complete set of gfp was constructed by the HiFi DNA Assembly Cloning Kit (New England BioLabs) with the F primer: 5′-gaattcattaaagaggagaaattaactatgagcaagggcgaagaactgtttacgg-3′ and the R primer: 5′-ctaattaagcttggctgcaggtcgacccttaatgatgatgatgatgatgtgagcctttatacag-3′. The gene of green fluorescent protein (GFP) was expressed under the control of the same promoter used for the ASKA strains. The E. coli AG1 strain, which is the host strain of the ASKA collection was purchased from Agilent Technologies .The ASKA (−) collection was obtained originally from the Coli Genetic Stock Center at Yale University. The no insert control of pCA24N was constructed by the Q5 Site-Directed Mutagenesis Kit , with the F primer: 5′-taagggtcgacctgcagccaagc-3′ and the R primer: 5′-atccgtatggtgatggtgatggtgagatcc-3′. The plasmid pCA24N-600nm = 0.15 with a total volume 150 µL of fresh LB media with 50 µg/mL chloramphenicol. Each plate had three biological replicates. The 96-well plates were sealed with oxygen-permeable membranes . The cell growth was monitored by reading the absorbance at 600 nm with microplate readers at 37 °C continuously. The doubling time was calculated by the equation: Doubling time = lg2/lgX. X is the growth rate in the exponential phase, which was automatically provided by Gen5 software designed for the BioTek microplate reader . The monitoring of GFP expression by fluorescence followed previous studies [Individual plates of the ASKA collection were replicated by inoculating 3 µL stock culture into 150 µL fresh LB media with 50 µg/mL chloramphenicol in each well of 96-well plates, and incubated at 37 °C overnight. The absorbance at 600 nm of each well was then read by the microplate reader. The overnight culture in each well was diluted to OD studies ,14.The software and online resources used for bioinformatical analyses were described in each subsection of Results and Discussion.d-1-thiogalactopyranoside (IPTG) for high-throughput growth tests. We monitored both recombinant protein production by the fluorescence intensity (600nm (p < 0.01 by the t-test), while commonly used concentrations of IPTG (0.5 to 1 mM) decreased the GFP expression significantly , leucine (Leu), and valine  are significantly increased in proteins which had severe effects on cell growth  of Ile, Leu, and Val. Most of the strains had improved growth, indicating that overexpressing proteins with high abundance of BCAAs could indeed impair cell growth, which could be then remedied by adding those BCAAs in growth media. b. We alsE. coli K12 proteins and in the group with severe growth effects was compared . For cellular localization, most of them are associated with membranes, which is consistent with the previous analysis of amino acid compositions, since membrane proteins tend to have higher abundances of nonpolar amino acids, including BCAAs due to their interactions with membrane lipids. Compared with the analysis of all the E. coli genes (p < 0.01 by the t-test). For biological processes, they span on all essential cellular processes. Compared with the analysis of all the E. coli genes (p < 0.01 by the t-test).For molecular functions, those proteins are distributed evenly in the three major categories: Enzymes, transporters, and binding proteins. Compared with the analysis of all the li genes , the frali genes , the frali genes , the frahttp://string-db.org) [Next, we analyzed the functional interaction network of proteins in the severe growth group by STRING database (-db.org)  (Figure asmB, lptD, lptG, dppC, lptF, and ftsQ. LptD, LptF, and LptG are three essential proteins in the lipopolysaccharide transport system [One cluster includes t system ,30,31. Dt system . FtsO ist system . AsmB ispheT, rplQ, rpsD, rpsG, and rnc. RplO, RpsD, and RpsG are components of ribosomes [Phe and editing activity [Another cluster contains ibosomes ,35,36. Ribosomes . PheT isactivity . Overexpactivity .bglH, mdtO, yciQ, yegI, and yihF. BglH is a carbohydrate-specific outer membrane porin [The last cluster includes ne porin . MdtO isne porin . YciQ isne porin . YihF anne porin . Again, ne porin .E. coli genes on its cell growth. Overexpression of membrane-associated proteins, or proteins with high BCAA abundances, tended to hinder cell growth. For recombinant protein production, it is suggested that the first thing is to check BCAA abundances, and supplementing BCAAs in growth media could recover cell growth when overexpressing proteins with high BCAA abundances. For membrane-associated proteins or proteins related to protein biosynthesis, it is recommended to reduce the rate of protein production with lower inducer concentrations, weaker promoters, or lower copy numbers of vector to improve cell growth for an increased total protein yields. In summary, we quantified the effect of overexpressing individual


The aims of this study were to (1) determine the association between diet quality using the Dietary Approaches to Stop Hypertension (DASH) score and cardiometabolic risk in a British working population and (2) identify employee characteristics associated with reporting a poorer quality dietary pattern.n = 5527) were included for sex-specific cross-sectional analyses. Dietary intakes were measured using 7-day food records. DASH score was calculated to determine diet quality. Logistic regression evaluated associations between (1) diet quality and increased cardiometabolic risk , and (2) poor diet quality (lowest fifth of DASH score distribution) and employee characteristics.British police employees enrolled (2007–2012) into the Airwave Health Monitoring Study  of increased cardiometabolic risk independent of established risk factors  and BMI: men OR 1.50 (95% CI 1.12–2.00), women: OR 1.84 (95% CI 1.19–2.97) compared to the healthiest diet group. Characteristics associated with reporting a poor quality diet were employment in Scotland vs. England: men OR 1.88 (95% CI 1.53–2.32), women: OR 1.49 (95% CI 1.11–2.00), longer working hours (≥ 49 vs. ≤40 h) men: OR 1.53 (95% CI 1.21–1.92) women: OR 1.53 (95% CI 1.12–2.09). For men, job strain (high vs. low) was associated with reporting a poor diet quality OR 1.66 (95% CI 1.30–2.12).The general population disparities in diet quality between England and Scotland were reflected in British police employees. The association of longer working hours and job strain with diet quality supports the targeting of workplace nutritional interventions.The online version of this article (doi:10.1007/s00394-017-1562-4) contains supplementary material, which is available to authorized users. Employment is considered beneficial to health and well-being when occupational hazards are controlled . Two typIt has been estimated that about a third of daily calorie intake is consumed in the workplace . TherefoThe two foremost food-based dietary patterns that have shown to be beneficial in the management of cardiometabolic disease risk are the Mediterranean diet and the dietary approaches to stop hypertension diet (DASH) . DietaryTo manage the increasing burden of cardiometabolic disease, public health strategies need to take account of wider environmental exposures as well as individual health behaviours. There is, therefore, a growing need to understand the relative influence of factors within different environments, such as the workplace, on dietary choice . Researcn = 15,404). The aims of this paper are to (1) measure the sex-specific association between diet quality determined Dietary Approaches to Stop Hypertension (DASH) score and cardiometabolic risk and (2) identify employee characteristics associated with reporting a dietary pattern associated with elevated cardiometabolic disease risk.The British police force employs over 250 000 men and women  across an = 322) as these diseases may affect cardiometabolic markers of interest. The present study sample (n = 5527) was comparable across key characteristics the total study sample  [We conducted a cross-sectional study using baseline data collected as part of the Airwave Health Monitoring Study between 2007 and 2012. The Airwave Health Monitoring Study was open to all police forces in Great Britain, recruitment procedures and baseline characteristics have been described previously . For the 42,112) . The AirDietary intake was measured using 7-day estimated weighted food diaries. Participants were provided with written instructions and requested to provide details on cooking methods, brand names and portion sizes. To aid portion size estimation, photographs based on those developed by Nelson et al. were provided . CalculaMean daily intakes were calculated for ‘positive’ and ‘negative’ food groups (g/day) and sodium (mg/day) for each participant  Supplementary Table S1 details the food group descriptions. The DASH scores were calculated as described by Fung et al. . Partici2). Waist circumference was measured between the lower rib margin and the iliac crest in the mid-axillary line using a Wessex-finger/joint measure tape .Enrolled participants attended a regional health-screening clinic. Trained research nurses used a standard protocol to conduct all clinical examinations as described previously . BrieflyFor each participant, cardiometabolic risk was estimated based on the presence (yes/no) of five established risk parameters: (1) central adiposity (waist circumference ≥ 94 cm for men and ≥ 80 cm for women), (2) dyslipidaemia , (3) elevated blood pressure , (4) inflammation (Hs-CRP ≥ 3 mg/L < 10 mg/L) and (5) impaired blood glucose control (HbA1c ≥ 5.7% and/or prescribed medication for glucose control). Increased cardiometabolic risk was defined as having three or more of these risk factors. Medication descriptions were matched to British National Formulary category and clinical indication determined.Information on occupational, lifestyle, medical history, socioeconomic and, demographic factors was collected during the health-screen visit using a structured on-line questionnaire. Total working hours  was classified into categories  based on previous research . Job dest tests were used for data with a normal distribution (mean and standard deviation presented) and Mann–Whitney U tests were used otherwise (median and inter quartile range presented). Associations across categorical variables were analysed using Chi-Squared test (χ2) with post-hoc comparisons conducted to determine the difference between subgroups [Ptrend) across the five ascending categories of DASH score (based on quintile ranges presented in Supplementary Table 3) and the association of having three or more cardiometabolic risk markers. Orthogonal polynomial coefficients (SAS function: PROC IML) were generated and applied to the contrast statement to correct for the unequal spacing between median values of each quintile of DASH score. Established confounding variables were included in the two models: (1) minimally adjusted: age (continuous), (2) fully adjusted: age plus categorical variables: physical activity, smoking status, education, TV viewing, job strain, menopause status (women only) and continuous variables: mean energy intake and mean alcohol g/day. Where a high level of collinearity was indicated between confounding variables  the variable explaining the greatest amount of variability was retained in the model. Due to the level of missing data for employment rank and work environment, these variables were not included in the multivariable analyses to maintain analytical sample size. Body mass index (BMI) was included in a separate model (continuous variable) as it potentially lies on the causal pathway between diet and markers of cardiometabolic health. Second, we used logistic regression to investigate the employee characteristics associated with consuming a poor quality diet (classified in the lowest DASH score group). Initially, we conducted bivariate logistic regression to determine the relationship between poor diet quality  with each covariate. We then adjusted for established predictors of diet quality based on previous studies . Odds ratios (OR) are presented with corresponding 95% confidence intervals (95% CI).All statistical analyses were undertaken using SAS version 9.3 . To assess differences between two groups, independent ubgroups . To corrp < 0.0001. There were significant differences between men and women for all occupational characteristics measured, with men more likely to be in higher rank , and have mobile job roles  and work 49 h or more per week . Women were more likely to be classified as having no markers of cardiometabolic risk compared to men (9.1 vs. 18.0%), while men were more likely to be classified as having three or more elevated risk markers (41.4 vs. 24.8%). Of those participants that had three or more risk factors, 84% men and 95% women, had central adiposity as one of the risk factors. Waist circumference was strongly correlated with BMI for men and women .The characteristics of the population sample by sex are shown in Table p < 0.0001) and non-milk extrinsic sugars , while men obtained more energy from alcohol . With the exception of whole grains and sodium, there were significant differences between energy-adjusted intakes across all food groups and dietary fibre, Table There were significant differences between sources of energy intake between men and women. Women derived more energy from carbohydrates , Table Logistic regression analyses showed a dose–response relationship across quintiles of DASH score and the odds of having three or more markers of cardiometabolic risk for men and women (Ptrend = 0.037).In bivariate logistic regression household income was associated with poor diet quality (lowest fifth of DASH score distribution) in men but not in women, though in men the negative association observed in the highest income brackets did not remain significant after adjustment for established predictors of diet quality . Advancing age was associated with reduced odds of having a poor diet as was being in the highest category for physical activity. The highest category of education (bachelor degree or higher) was negatively associated with poor diet quality, and remained significant after adjustment . For men and women geographical region and smoking status were positively associated with poor diet quality , Table The present study demonstrated that diet quality determined by DASH score to be negatively associated with cardiometabolic risk within British police force employees. Employees in the lowest fifth of DASH score distribution had increased odds of cardiometabolic risk independent of other lifestyle factors and BMI. This study has also identified that a poor dietary score was also associated with other negative lifestyle behaviours . Furthermore, employees reporting long working hours and high job strain (for men) had increased odds of reporting a dietary intake that was associated with cardiometabolic risk.Previous studies in German and US police officers have observed the prevalence of metabolic syndrome to be higher in this occupational group compared to the general population , 25. In There were clear dietary differences across men and women, with female police employees more likely to report a diet with a higher concentration of fruit and vegetables, and lower concentration of processed and red meat. This observation supports a previous study that also reported that women more likely make healthier choices (higher fruit and vegetable intake with lower intake of high fat foods) compared to men . The difDespite DASH being commonly applied to US cohorts and its recommendation by the American Heart Association, to date the DASH score has not been widely applied to UK cohorts to determine cardiometabolic health . In the n = 247) observed that fruit and vegetable intake was positively associated with physical activity [In agreement with previous studies in European populations, we observed advancing age  and highactivity . In the activity .We observed that the number of weekly working hours had a positive association with odds of recording a poor quality diet a relationship that was independent of age, education and region of employment for men. Previous studies investigating dietary intake and number of working hours have mainly used food frequency questionnaires or food surveys to collect dietary data –40, therThe strengths of the present study are the large-scale collection of 7-day estimated weighted diet records from a young single occupational group who are potentially at higher risk of cardiometabolic disease compared to the general population , 25. TheIn conclusion, this study has profiled the dietary intakes and measured the diet quality (using the DASH score) of British police force employees. The novel aspect to this study has been the application of 7-day food diaries in the investigation of occupational exposures and diet quality. In general, the observed differences in diet quality across different employee characteristics  in the police force reflect those within the general British population. Our findings suggest that for male police force employees longer duration of weekly working hours and higher job strain are associated with reporting a diet quality indicative of increased cardiometabolic risk. These findings potentially support the suggestion that dietary differences may contribute to variation in cardiometabolic disease risk observed across employees working longer hours or with high job strain. To test this hypothesis, future longitudinal studies are required to investigate if diet quality meditates the relationship between these occupational exposures and cardiometabolic disease. Additionally, further investigation to examine the barriers to healthy eating amongst male employees working extended hours or with high job strain is warranted. Gaining an understanding of why this cohort of employees are more likely to consume a diet associated with increased cardiometabolic risk will assist nutritional and occupational practitioners in targeting workplace nutritional interventions.Below is the link to the electronic supplementary material.Supplementary material 1 (DOCX 71 KB)


The maternal mortality rate in Lao PDR (Laos) is still the highest in Southeast Asia, at 197 per 100,000 live births. Antenatal care (ANC) could contribute to maternal and child mortality reduction. The quality of ANC service remains inadequate and little information is available on the quality of health education and counseling services of health providers in Laos. This study aims to gain insight into the perceptions of stakeholders on both supply and demand sides of public ANC services in Laos and evidence for recommendations to improve the quality of ANC services.Semi-structured interviews were conducted with 50 participants from different stakeholder groups; on the demand side, couples with a currently pregnant woman and mothers with children under one year of age and a family member; and on the supply side, health providers, managers, policy makers of the Ministry of Health, and development partners. The interviews were voice recorded and transcribed verbatim for analysis by open and thematic coding, using the MAXQDA software program.All respondents reported that the number of pregnant women who visit ANC services has increased. However, an analysis of the supply side identified issues related to the quality of ANC that need to be improved in the areas of facilities, human resources, privacy and confidentiality, providers’ behavior, attitudes, and ineffective communication skills when it comes to providing health education and counseling to pregnant women and their family members. The analysis of the demand side mainly emphasized the issues of providers’ behavior, attitude, communication and unequal treatment, and the lack of privacy. Both sides also suggested solutions to the problems, such as training, effective materials, rewarding good role models, and building a feedback system.The number of public ANC services has increased, but both supply and demand sides experienced challenges with the quality of ANC. All respondents proposed possible solutions to improve quality of ANC service in public health facilities in Laos. Maternal and child mortality and morbidity are major health concerns worldwide . Ninety-The WHO prepared a framework for improvement of quality of care that requires effective provision of care and positive experience of care; it includes supply of competent, motivated staff and essential physical resources , 5. A syEffective pre-, intra-, and post-natal care interventions play an important role in reducing child mortality and morbidity –10. ReceThe quality of ANC services influences women’s healthcare seeking behavior , 13. EffIn Laos, the quality of ANC provision in general remains inadequate. However, little is known about the health education and counseling given to clients during ANC consultation. Previous studies suggested that ANC quality was poor in rural areas due to the lack of equipment and materials and weak health care provider skills , and thaThis study aims to fill that gap, by looking into the quality of communication, specifically health education and counseling in the context of ANC in Laos. Stakeholders at all levels in the health system, from central to community, were invited to give their views. The specific objectives were 1) to explore the provision of care and experiences from supply and demand sides with reference to a quality of care framework that included routine care, providing information, effective communication, respect and dignity, emotional support, competence, motivation, and essential physical resources, and 2) to gain perspectives of different stakeholders on ways to improve the quality of ANC in public healthcare facilities in Laos. The results of this research could provide health policy makers and planners with insights from practice to improve the quality of ANC services particularly in the development and implementation of the new Lao ANC guidelines.This was a qualitative study with semi-structured interviews, conducted from April to July 2017 with key informants from different stakeholder groups using guidelines with open questions. At the same time as the interviews, we made on-the-spot observations of the healthcare facilities we were visiting.We identified four stakeholder groups for this study. On the supply side: 1) directors and academic staff from each of the Department of Hygiene and Health Promotion, the Nutrition Centre, and the Information Education and Communication (IEC) Centre of the Ministry of Health (MoH) currently working in the area of nutrition, maternal and or child health; 2) health providers currently practicing at the ANC services, with at least one year’s experience in providing care to pregnant women, in the central, provincial, and district hospitals, as well as health centers; 3) representatives of development partners such Save the Children, WHO, UNICEF, UNFPA, Plan International, and World Food Program, staff currently working in the area of nutrition and/or maternal and child health; and on the demand side: 4) currently pregnant women, mothers with children under one year of age, and their family members (parent/husband/grandmother).Study sites were selected using both purposive and random selection. Vientiane Capital was purposively selected as the location of policy makers and development partners as well as the central level hospitals. Of the four provinces  in Southern Laos, where ANC services have been developed to a similar standard , two wern = 16); four couples of Lao Loum group, which is the majority ethnic group in Laos (two couples with currently pregnant women and two couples with mothers having children under one year of age) and four couples of an ethnic minority group (two Laven and two Brao (Lavae) couples with currently pregnant women, and mothers with children under one year of age) , one expatriate Master student (MJ), and two field research assistants. The interviewers were provided with a three-day training course prior to data collection. The interview guides were piloted in the central and provincial levels with two MoH staff, four health providers and two mothers, which led to reformulation of several questions to finalize the guides.Before the interview started, the interviewers explained the aim of the study and the general topics that would be discussed. They were told that the records would be anonymous and that they could withdraw at any moment without giving a reason. Written consent was obtained before each interview. Most of the providers were interviewed in English by MJ with the support of the local academic researcher (SP) who translated when needed. The service users were interviewed directly in the Lao language by SP. All interviews were recorded. The time of interviews ranged from 32 to 113 min (average 68). Notes were made while conducting the interviews, for the summary afterwards. During the interview, spot observational field notes were made which were included in the summary of the interview.The recordings and interview notes were used to generate verbatim written transcripts for the data analysis, which were analyzed using the software program MAXQDA. The research team read the transcripts a few times and discussed the open coding process together to reach consensus on the code tree. Then, using the concepts of the quality of care of WHO framework , 5 all mThe findings related to provision of care, experience of care, essential physical and human resources, staff competence, and outcomes, are described based on the concepts of quality of care as defined by WHO . In TablI think some young staff are not able to provide ANC service properly. They cannot identify the status of the fetus.” “The majority of participants mentioned that health providers could not always perform routine care correctly, which may be partly attributed to the lack of training, materials, and guidelines. For example, providers mentioned that (young) staff may have to perform their tasks without having had any specific training, so they lacked competence to measure the position and heartbeat of the fetus when performing obstetrical examination, or were not trained to provide health education on nutrition, physical activities, and danger signs for health care seeking during pregnancy and postpartum period, as illustrated by the quote below.“I was not very happy when I visited the health center. The doctor did not provide me any medicine, but told me to buy it outside at a drug store. So it is better to go to district hospital and provincial hospital.” There were different understandings of how routine care should be provided, which can be seen from the diversity of guidelines observed as present in facilities. A few facilities developed their own clinical guidelines or SOPs, when a national standard guideline was not available. Another issue in routine care was a lack of essential medicines. For example, several respondents on the demand side mentioned that the health facilities had no medicine for them.Some providers did not know how to explain properly to women and family members, due to lack of training and materials. I myself have never been trained for health education and counseling.” “The respondents of both supply and demand sides alike mentioned that effective communication, especially in relation to counseling, was far from optimal. The main reasons given were similar as mentioned above: lack of time, guidelines, materials, and training. Counseling is not a stand-alone activity and is often given during physical examination. It was characterized by one-way provision of information rather than a mutual communication process. In some cases, health education was given to a group of several women and family members together at one time.I only accompanied my wife to the hospital and just waited outside the room. When I asked my wife about what the doctor told her, she said that the doctor talked very little and quickly during the physical exam, then asked her to go out, and she did not know and could not remember what the doctor talked about.” “All health providers mentioned that time limitation did not allow them to provide sufficient information, because other clients were waiting outside. Some providers felt shy to speak with the women too much, because it is not their cultural habit to do so. Some providers did not ask the women about their needs, because they were afraid of not being able to fulfill the needs, which would be unacceptable for them. Service users also reported that very limited information was provided, and only to women – not to their family members, as illustrated by the following quote:“Some clients may want to talk more, but sometimes they are afraid that if they talk more to the provider, he/she may not provide very good care to them. So they only talk about small issues even if they have a problem.” In some situations, providers explained that it is not their culture to have a discussion. Women see the doctor as an authority that should tell them what to do because of their knowledge. In addition, some healthcare providers noticed that even if women did have the opportunity to ask a question, they remained silent. Also, although it does not seem to be a hierarchical issue, the clients are sometimes reluctant to discuss their health problems with health providers since they may be afraid that the provider may become angry with them if they ask (too many) questions.“The issue is that they do not do counseling, while counseling is needed to bridge the gap between telling someone what to do or having a slogan or poster, and actually helping them [with tailor made advice] to achieve change.” According to development partners, participants at the MoH and some providers, when women do not ask questions or explain their personal issues to the healthcare providers, the provider cannot give appropriate counseling to the needs of the client. The development partners and the participants from the MoH all assumed that this way of providing information would result in better outcomes for both mother and newborn.Inadequate behavior and attitude of health providers was described as reflecting the problems of poor respect, lack of dignity and emotional support for users of ANC. Most participants mentioned that negative interpersonal interactions between health providers and service users occurred at all health facilities. For example, some providers were aggressive, unwelcoming, impolite, and unfriendly, spoke too loudly, used rude words, and did not smile to women and family members. Some service users mentioned that they experienced inappropriate behavior and negative attitude of health providers at least one time when they accessed ANC services in their community.“I used to go for ANC at the health center; the staff blamed me that my body was unclean, smelly , and shouted at me that next time I must take a bath before coming. The second time, when I went to district hospital for my child’s vaccination, a health provider scolded me, asking “where were you when our staff went to provide vaccination in the village?” I was scared and sad; I am a poor person but do not wish to meet this kind of person.” Some participants of the supply side tried to explain the reasons for such inappropriate behavior and negative attitude of health providers. Poor working environment resulting in stress, fatigue, frustration and poor job satisfaction were mentioned. These were caused by high workloads with long working hours, lack of supportive supervision, and insufficient salaries without additional incentives. Most providers indicated that they and their colleagues do not always comply with behavioral norms, such as speaking in a soft voice and properly greeting clients. Clients indeed indicated that some providers talked very loudly and this was seen as very inappropriate.lack of privacy. It was observed, and confirmed in interviews, that it is common that several women were examined in the same room at the same time, at all levels of health facilities. Several service users also mentioned that sharing a room for physical examination was not appropriate, and they felt very ashamed if other people saw their bodies.“I felt very ashamed during physical examination, because other women were in the same room with me, and the window was also open, so people outside the room might see my body.” Another issue was the Furthermore, people in waiting areas could easily overhear conversations held in the examination room. For example, the windows towards the waiting area were open, there were no closed walls between the different rooms of the ANC services, and in some facilities, and these services were provided in the waiting room itself.“I hesitated to visit ANC because I was afraid of paying more money to the provider. If I did not pay additional money or bring something to the health care worker, she may not treat me as well as she did others, especially her own family, those she knows well, and the richer people.” Moreover, most service user respondents reported unequal treatment, mainly at the lower level of health facilities, as a challenge. For example, providers paid more attention and were more polite to their own relatives, to more affluent users and to those who paid extra money to them.The majority of the respondents clearly felt that poor provision and experience of care is caused by a number of factors concerning weak physical and human resources, including staff numbers, staff competence, materials, medicines, and physical space. Supply-side participants suggested four reasons for lack of (competent) staff: 1) experienced staff was assigned to work at another ward; 2) qualified staff volunteers to move where better incentives are available; 3) experienced staff was retired, and 4) new staff lacks experience and specific training.Supply-side participants also noted the lack of specific health education materials and standard guidelines to be used at health facilities at all levels, possibly explained by problems with financial support, distribution mechanisms and development. For example, many participants mentioned that materials were sent from central to provincial level, at government expense, but then the provincial level has to send it on to district hospitals and health centers without any financial support. So materials that should have been available did not reach health providers at the lower levels. Several participants, particularly IEC academic staff, reported that most existing IEC materials were developed with support from development partners or government. IEC centers themselves have no specific budget to develop specific materials and were not involved in the development of clinical guidelines.“Our service is better than it has ever been; we have the rooms and place to provide the ANC service. Also the number of pregnant women who visit the ANC increased recently in our hospital.” Although we did not specifically ask about health outcomes, most participants referred to outcome indicators when discussing the quality of care. According to most participants from the supply side, utilization of ANC services has recently increased:However, some respondents, on supply and demand sides alike, stated that if the overall quality is not adequate, this increased utilization is not going to be sustainable.Abundant suggestions were made about how to improve the quality of ANC service with respect to inadequate provision of care, poor experience of care, and inadequate human and physical resources. The most relevant proposals are discussed based on the responses of the study participants and how these are linked to the problems noted above.“Besides integrating communication skill into the curriculum in medical school, I think we must continue to provide medical ethics training to health care providers, to repeat and remind them, because the existing curriculum of the university may not be complete.” .All participants on the supply side suggested that training would be the first option to improve the health staff competency and increase awareness of medical ethics. For example, short course training, particularly for new staff, on routine care and effective communication skills would help to refresh health providers’ knowledge and skills. Also they strongly recommended that refresher training on medical ethics should be done at least once a year for health providers at all levels, which might help to reduce the problems of lack of privacy/confidentiality and unequal treatment in the public health facilities. In addition, they also suggested that there is a need for more substantial, long-term training. For example, effective communication skills, particularly counseling skills and a comprehensive medical ethics course, should be integrated into the curriculum of medical schools, and offered as extra-curricular courses to providers, as one health manager said.“I think other pregnant women would also like to get very good material with clear and very nice pictures to bring home to tell family members; for myself, I would get more support from them about my health and about child care.” Besides the training, nearly all participants on both supply and demand sides suggested that national standard guidelines and effective materials should be available to health providers at all levels, to help provide more effective ANC services with a constant standard. Participants of both sides specified that effective materials must be clear, with attractive pictures, for both health providers and service users. Many participants, the IEC staff in particular, proposed that the design of materials should involve the clients or health care users as well as providers, from community level up. Most participants also suggested providing specific material for clients to bring home to increase the knowledge of service users, and possibly to influence other people living in the same family and community. For example, many demand side participants mentioned that it would be good if women could be given effective materials that they could share with others, particularly husbands and elderly people who have more authority in their communities.“One way is to learn from colleagues abroad, specifically healthcare services in neighboring Thailand, which has similar culture and language, that would be the most effective way to increase health providers’ motivation to act differently.” All participants on the supply side mentioned that having good role models would be one of the most important ways to change health providers’ behavior and attitudes. For example, health providers could learn from and exchange experiences of best practices with other health providers, which might help them to gain more emotional control and increase confidence. Most participants on the supply side suggested that a good role model could be a person in their own health facilities, from other provinces, or even neighboring countries.Additionally, the majority of respondents strongly advised that one of the most important solutions, with low cost and simple implementation, would be a peer-feedback system, for example, a routine (weekly or monthly) feedback system among providers. There should also be individual feedback immediately by the manager. A feedback box should be more available and accessible for service users to give their feedback.The study aimed to analyze the current quality of ANC provision within the public healthcare system in Laos and ways to improve it, with a focus on the need for good health education and counseling. This was done by exploring perspectives of a range of supply and demand side actors using the WHO’s Quality of Care Framework . We founfree delivery vouchers for maternal and child health care [Although ANC utilization has recently increased, there are serious issues with quality of care, which was, without hesitation, made clear by both health providers and policy makers and confirmed by reports of service users. The increase in use can likely be attributed to the recent policy implementation of lth care , 22. Thelth care . In anotlth care , howeverThe results indicated that essential physical and human resources remain poor in most health facilities at all levels. These findings are consistent with other research reports , 25, 26 A lack of incentives for healthcare providers in their routine ANC service was identified as another major concern in most health facilities, which is found elsewhere, especially in China, where it was recently revealed that lack of incentives for health staff is a major concern, because they could not afford to increase payments for the staff . A systeIn addition to inadequate job performance skills, the unavailability of materials and healthcare guidelines limited effective performance by health providers . Having Most respondents felt that poor staff competencies and the limited attention to medical ethical issues might be improved by training. The “Health Sector Reform Strategy and Framework” also stated that increasing staff competency and medical ethics through training helps to improve quality of care . Taking Many study participants suggested that having role models of good practices is probably one good way to improve the interpersonal interactions between health providers and health users. Appropriate behavior and a positive attitude of health providers could become the normal practice at health facilities when health workers follow a good role model. Friendly, respectful and caring health providers can encourage health care utilization and client satisfaction . It woulAlthough ANC service users mentioned that they experienced inappropriate behavior of health providers, both supply and demand sides might not be aware of the full potential of the negative effect of that problem. A study in Hong Kong demonstrated that emotional or psychological abuse during pregnancy resulted in a greater risk of postnatal depression, higher risk of thinking of harming themselves, and significantly poorer mental health-related quality of life . TherefoIt is well-known that a good quality of care plays an important role in reducing child mortality and morbidity . Recent Since this study was conducted within the framework of a research to policy program, the specific information needs for ANC guideline development were taken into account. Additionally, future medical ethics and communication trainings are planned to be developed and integrated into the curriculum of the University of Health Sciences, Lao PDR.Since this qualitative study was only carried out in the two southern provinces of the country, we cannot generalize the findings to the whole country. However, according to the perceptions of the respondents from the central level, many issues arising in this study would be similar in other geographical areas of the country. Another issue is that some participants were interviewed in English with questions and answers translated back and forth between English and Lao, which could limit natural conversation between interviewers and interviewees. Respondents at community level were interviewed only in Lao, while some of them were from another ethnic group whose mother language was not Lao. However, dialect translation was done in some cases. All respondents appeared to express their opinions openly and recordings ensured that all information was collected for analysis.Although utilization of ANC services in Laos has recently increased, the quality of ANC service remains inadequate. The following actions should be taken by the Lao government: investing in the development of effective IEC materials for counseling and ANC guidelines; training on ANC routine care, counseling and medical ethics; increasing motivation for health care providers by providing direct incentives; and essential physical resources . Additionally strengthening the feedback system and building exposure to good role models into both pre-service and in-service training are strongly recommended. Additional information to deepen understanding of the communication issues could be gained from a participant observation study and program implementation.


In this work, the physicochemical properties and in vitro bioactivity and cellular viability of two commercially available bovine bone blocks  with different fabrication processes (sintered and not) used for bone reconstruction were evaluated in order to study the effect of the microstructure in the in vitro behavior. Scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectrometry, mechanical resistance of blocks, mercury porosimetry analysis, in vitro bioactivity, and cell viability and proliferation were performed to compare the characteristics of both allograft materials against a synthetic calcium phosphate block used as a negative control. The herein presented results revealed a very dense structure of the low-porosity bovine bone blocks, which conferred the materials’ high resistance. Moreover, relatively low gas, fluid intrusion, and cell adhesion were observed in both the tested materials. The structural characteristics and physicochemical properties of both ceramic blocks (sintered and not) were similar. Finally, the bioactivity, biodegradability, and also the viability and proliferation of the cells was directly related to the physicochemical properties of the scaffolds. In severely atrophic jaws, reconstructive bone surgery needs block grafting, especially in cases where the resorption of the edentulous maxilla can create a reverse maxillomandibular relation or an increased vertical distance between the jaws . In thesGiven the morbidity associated with using autografts, there is growing interest in developing animal or synthetic bone graft substitutes ,2,3. AltA good bone substitute should exhibit a similar morphology to a lost bone by displaying an internal architecture that allows the perfusion, adhesion, growth, and maturation of cells. It should also be highly hydratable to maintain an isosmotic environment that allows suitable mobility and growth of angiogenic factors, as well as low immunogenicity and being able to be fixed surgically with screws or wires. Besides that, the bone substitute should also be absorbed during a time that is compatible with bone formation ,10,11. FHydroxyapatite ceramics manufactured from natural materials, such as coral or bone, shows the advantage that they inherit some properties of the raw materials such as the pore structure ,15,16. THowever, the process of treating materials for bone grafts, and their ideal composition, have been discussed and are subject to discrepancies. For example, the sintering process can either be used or not when preparing bone grafting substitutes to promote physicochemical effects in these materials ,16,21. CSome authors studied five commercially available bone graft blocks, DIZG (Advancing the world of tissue transplantation) Human-Spongiosa, Tutobone, Puros Allograft Spongiosa, OsteoBiol Sp, and Bio-Oss were histologically studied. Three out of the five bone blocks contained organic/cellular remnants, as revealed by histology stated that their blocks were free of such remnants. These remnants could result in an unexpected and undesired immune response, inducing a foreign body reaction, and thus compromise the safety, biocompatibility and function of these three bone substitute materials . TutobonWe compared these materials with a synthetic tricalcium-phosphate (TCP), evaluated as a negative control of the analysis process, by analyzing the porosity, chemical, and mineralogical composition and sample microstructure. Hence, the objective of the present study was to evaluate the effect of the microstructure , on the in vitro behavior of the two commercially available mineralized bovine bone blocks prepared by two different processes: sintered and not sintered, in a simulated body fluid, and the differential cell proliferation of a mouse preosteoblastic cell line (MC3T3 cells) growing on these surfaces.®  called bovine bone 1, treated by a physicochemical process, e.g., the material was subjected to high-temperature deproteinization for sintering (950 °C), treated with organic solvent, and sterilized  + Ca2+ + 2OH− → Ca10(PO4)6(OH)2, and leads the nucleation on the surface of the globular particles of Ca–P powders that by that time transforms into a new Ca–P rich layer. This precipitate was growing with exposure time . The bovine bone 1 with an intermediate behavior showed some precipitation, but the dissolution of the material was still the dominant process (1.2% of the weight was lost). This behavior can be correlated with the crystallinity of the materials.The precipitation process was dominant in high crystalline a material and the dissolution process will be dominant in materials with poor crystallinity. The mechanism of bioactivity is a competitive process of dissolution–precipitation where it is possible to model the behavior of the materials changing the physicochemical characteristic of the graft materials through the manufacturing conditions, so we can design materials with specific needs.The biological behavior of the materials can be studied by cells cultures over your surface. In this way, several researches have been carried out to investigate the interaction between cells and biomaterials ,52. In tWithin the limitations of this study and in view of the results obtained, we cannot say which of the two materials (sintered or not sintered) is better. Each material can be used in a specific clinical application in reconstructive surgery of bone pathology. Bovine bone 1 (sintered), which represents the moderate dissolution pattern, allows implementation in situations requiring partial replacement by autologous bone with a matrix in place over the longer time. Bovine bone 2 (not sintered) with a faster dissolution rate allows its use in situations requiring rapid replacement by autogenous bone. The control material with of slow resorption pattern is suitable in the situation where dimensional stability of the implant is required.In conclusion, the data from this study reveal that variations in physical properties, like phases, crystallinity, and porosity of calcium phosphate-based materials of either a natural or synthetic origin, strongly depend on manufacturing procedures. The bovine bone materials are monophasic with high porosity and medium-high crystallinity, except for bovine bone 2, whose crystallinity is lower due to the presence of a bigger quantity of collagen mixed with the hydroxyapatite matrix. As expected, the best crystallinity corresponds to the synthetic material. Pore size is similar for the studied materials of a natural origin, with certain advantages for the synthetic TCP material in relation to external pores and micropores interconnection. The viability and proliferation of the cells growing in direct contact with the bovine bone blocks are relatively low in comparison to the control group.


Global usage of electronic nicotine delivery systems (ENDS) has been increasing in the last decade. ENDS are non-combustible tobacco products that heat and aerosolize a liquid containing humectants, with added flavorings and often nicotine. Though ENDS are promoted as a less harmful alternative to smoking, current evidence links their use to a wide range of deleterious health effects including acute and chronic lung damage. ENDS can elicit an inflammatory response and impair the innate immune response in the lungs. Exposure to ENDS flavorings results in abnormal activation of the lung epithelial cells and β-defensins, dysfunction of the macrophage phagocytic activity, increased levels of mucin (MUC5AC) and abnormal activation of the neutrophilic response (NETosis). ENDS menthol flavorings disrupt innate immunity and might be associated with allergies and asthma through activation of transient receptor potential ankyrin 1 (TRAP1). Recent studies have expanded our understanding of the relationship between the homeostasis of lung innate immunity and the immunomodulatory effect of the host-microbiota interaction. Alterations of the normal respiratory microbiota have been associated with chronic obstructive pulmonary disease (COPD), asthma, atopy and cystic fibrosis complications which are strongly associated with smoking and potentially with ENDS use. Little is known about the short-and long-term effects of ENDS on the respiratory microbiota, their impact on the innate immune response and their link to pulmonary health and disease. Here we review the interaction between the innate immune system and the respiratory microbiota in the pathogenesis of ENDS-induced pulmonary dysfunction and identify future areas of research. Global usage of electronic nicotine delivery systems (ENDS) has increased in the last decade, especially among youth and young adults . The preThe recent outbreak of electronic cigarettes, or vaping, product use-associated lung injury (EVALI) raised national concern about the harmful effects of ENDS products and constituents . SimilarThe respiratory microbiota has also shown to play a crucial role in the lung innate immunity and inflammatory response in health and disease . Recent Prevotella spp.), Firmicutes (including Streptococcus spp. and Veillonella spp.), Proteobacteria, and Actinobacteria  and viral communities (Anelloviridae family)  .Tropheryma whipplei in the LRT  microbial immigration from the oropharynx  ; (b) mic the LRT  and the  the LRT . A unifi the LRT .The airways epithelium provides a biophysical protective barrier  and a siHaemophilus species in patients diagnosed with COPD. Thus, the respiratory microbiota interacts with the airway epithelium and phagocytic cells in a positive feedback loop to develop immunological tolerance and prevent exaggerated inflammatory responses. Further research is needed to elucidate specific mechanisms explaining immunological tolerance in the respiratory tract and to identify potential intervention targets in respiratory pathologies.According to The production of mucus and antimicrobial peptides and proteins is a key element of the innate immune system of the lung. The airway surface liquid facilitates the mucociliary transport of toxicants, microorganisms, or particulates; lubricates and hydrates the airways; and, contributes to the epithelial barrier . The muc2, immunoglobulin A, lung, and nasal epithelium clone protein, collectins (surfactant protein A and surfactant protein D), mannan-binding lectin, cathelicidins, and β-defensins  in the airways (Nicotine has been shown to increase mucin production and mucus viscosity through activation of the α airways . Nicotin airways . However airways . Inhaled airways . Further airways . Thus, nin vitro model. Moreover, ENDS flavoring chemicals play a major role on the cytotoxic effects of e-liquids and e-liquid aerosols . ExposurStaphylococcus aureus, Streptococcus epidermidis, and Micrococcus luteus (Additionally, Clapp et al. showed that exposure to cinnamaldehyde-containing ENDS might decrease the mucociliary escalator motility by reducing bioenergetic activity  and may s luteus  and membs luteus . ENDS mes luteus , arthrits luteus , and asts luteus . Though in vitro, animal, and human studies are increasingly showing a link between the pathophysiology of respiratory disease and the exposure to ENDS aerosols (Little is known about the short- and long-term effects of ENDS on the respiratory microbiota, their impact on the innate immune response and their link to pulmonary health and disease. Preliminary studies have expanded our understanding of the relationship between the homeostasis of lung innate immunity and the immunomodulatory effect of the host-microbiota interaction. Existing and emerging evidence from aerosols . Relatedaerosols , asthma aerosols , and cysaerosols , which aFew studies have focused on the relationship between the respiratory microbiota and the immune response to air pollutants and toxicants in the respiratory tract. in vitro and in vivo models for the realistic evaluation of the respiratory microbiota-lung innate immune response in the context of immunotoxicity studies. Further research on the pathogenesis of flavor-vaping induced lung injury must consider the respiratory microbiota as a potential mediator of the immune and the inflammatory response after exposure to toxicants.The recent EVALI outbreak in the US, coinciding with a surge in youth vaping, generated a movement in the research community to establish research priorities and address challenges to support the Food and Drug Administration and World Health Organization efforts to regulate and control ENDS . ElucidaZQT, DJO, and IR initiated the idea for writing the manuscript, reviewed the literature, and edited the manuscript. ZQT drafted the manuscript. ZQT, DJO, IR, DC, DL, and SG revised, edited, and approved the final version of the manuscript.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.


Does the inclusion of a randomized inter-trial interval (ITI) impact performance on an Attentional Blink (AB) task? The AB phenomenon is often used as a test of transient attention ; howeverMage= 21.5, SD = 7.4) completed both versions of the task, in counterbalanced order.Thirty university students (21 female; age range 18–57, No significant difference in performance was found between the standard AB task and the AB task with the random ITI. Bayesian analyses suggested moderate evidence for the null.Temporal unpredictability did not appear to impact task performance. This suggests that the standard AB task has cognitive properties with regards to task difficulty, engagement, and motivation, that are inherently similar to tasks that employ a randomized ITI to measure sustained attention (e.g., the Psychomotor Vigilance Task; PVT;  Given the important role attention plays in behavioural outcomes , many reThe AB occurs when an observer has a reduced ability to perceive a second target (T2) within a set of distractors when it is presented within 800 ms of an initial target T1; see . The phePast research has investigated many factors thought to influence performance on the AB task, including individual differences . AlthougIn the context of AB, foreperiod refers to the time between trial commencement and presentation of T1. The impact of foreperiod on AB was investigated by lag 1 sparing, a robust feature of the AB  refers to the length of time in milliseconds between the presentation of T1 and T2. In AB tasks this is commonly referred to as the “lag”, which denotes the number of distractors between T1 and T2. Lag is arguably the strongest predictor of T2 perception: T2 is reliably perceived when presented immediately after T1 , making it resistant to the AB effect. This is known as f the AB . The abif the AB .Introducing a consistent lag or TOA  improves AB task performance, especially when participants are made explicitly aware of the consistency . Even whITI refers to the time between a participant’s response on a trial N−1, and the start of the next trial N. Although AB tasks classically employ a fixed ITI, the present research seeks to explore the role that the ITI plays in shaping task performance. The time between one trial ending and the next trial commencing may be crucial with regard to preparation and anticipation, and as yet these intervals have not been investigated in relation to the AB. This would be particularly relevant to research investigating the role of pre-stimulus alpha .Although historically it has been assumed that ITI does not affect AB performance, ITI has been found to influence both reaction time and accuracy in vigilance tasks, such as the Psychomotor Vigilance Task PVT;  and the increases the AB by reducing temporal predictability, thus lessening readiness for the task.Previous studies investigating the temporal predictability of the AB have manipulated temporal attributes within the RSVP stream. Usually this is achieved by identifying a temporal aspect which is typically variable , and applying a fixed duration, to see if this attenuates the AB. However, the current research seeks to investigate this by taking a temporal attribute of the task which is usually fixed (ITI) and varying it, to see if greater variability Mage = 21.5, SD = 7.4) provided written informed consent and participated voluntarily. Most were psychology undergraduates who were received course credit. Ethical aspects of the research were approved by the Australian National University Human Research Ethics Committee (protocol 2015/184), in accordance with the ethical standards of the 2008 Declaration of Helsinki.Thirty observers  with a refresh rate of 120 Hz and a resolution of 1,920 × 1,080 pixels. Behavioural responses for the AB were collected using a Cedrus RB-8 SUBJECT response box, with buttons labelled 2, 3, 4, 7, 8, and 9. Two buttons were left blank to enable participants to indicate if T2 did not appear or they did not perceive it. Experimental stimuli were created using Psychophysics Toolbox (version 3) for MATLAB R2012b .2) on a grey (17.3 cd/m2) background. Items were presented using an RSVP stream in 1.23° Helvetica font. Each trial commenced with a fixation cross and contained between 18 and 22 stimuli. Targets were numerals  and distractors were uppercase letters, excluding those that could be confused with numbers . Each stimulus in the stream appeared for exactly 100 ms (12 frames at 120 Hz). Most trials included two targets (T1 and T2), with some ‘blank’ trials including only one target. T1 appeared with a jitter of ±2 items, after 4-8 distractors, and T2 was presented at either lag 3 or lag 8. In both conditions, 30 trials were presented for each lag, with lags randomly interleaved. When each AB trial had concluded, participants were allocated 3.5 s in which to respond. If participants did not respond within the time frame, a non-response was recorded, and the next trial commenced. The ITI commenced immediately after the participant had keyed in their answer for T2, or after the time limit had elapsed.Stimuli were displayed in white .During the Participants were given two AB practice blocks with auditory feedback. Both practice trials used a fixed ITI. A high-pitched beep indicated that the participant gave the correct response, while a low-pitched beep indicated that the participant gave an incorrect response. The first practice block involved 12 trials at half speed (24 frames per second). The second practice block was at full speed. Following this, participants completed the two experimental blocks in counterbalanced order. Experimental trials took place in a darkened room to avoid peripheral distractions. To ensure consistency and minimise distraction, the experimenter was not present during the experimental trials. After completing both versions of the task, participants were asked verbally if they had detected any difference between the two versions of the task. Qualitative verbal responses were recorded. After answering this question, participants were verbally debriefed.lag8 − T2|T1lag3), and was compared between fixed-ITI and random-ITI conditions. T1 accuracy was also compared between conditions, to ensure that the blink magnitude results could not be attributable to systematic differences in T1 identification. Statistical analyses were conducted in jamovi, version 9.5.15 , which in simple terms gives an odds ratio of how much more likely the null is than the alternative hypothesis  for the fixed-ITI condition and .87  for the random-ITI condition. A Wilcoxon Signed Rank Test indicated there was no systematic difference in T1 accuracy between conditions, W = 167, p = .829, M (diff) = .01, SE (diff) = .03, 95% CI [−.04–.03], δ = .07. In addition, we carried out a Bayesian paired-samples t-test analysis using JASP  = .03, 95% CI [.11–.23], δ = 1.02, and the random-ITI condition, W = 446, p < .001, M (diff) = .15, SE (diff) = .03, 95% CI [.10–.21], δ = 1.02.t-test  indicated that blink magnitude did not significantly differ between fixed-ITI (M = .18 SD = .14%) and random-ITI  blocks, t(29) = .49, p = .631, M (diff) = .01, SE (diff) = .03, 95% CI [−.04–.07], δ = .09. A follow-up Bayesian paired-samples t-test using JASP showed moderate evidence for the null, using a Cauchy prior of .707, BF01 = 4.61, error % = .007.A paired F = 0.3, p = .617, η2p = .009, or order, F = 1.7, p = .202, η2p = .057, and no significant interaction between trial order and ITI, F  = 3.5, p = .072, η2p = .11. However, it should be noted that this analysis had low power, as it was not initially included as part of our experimental design, so would be worth exploring in further research. The results are plotted in As the trial order was randomised (either random ITI first or fixed ITI first), order effects were analysed in the results using a mixed ANOVA on blink magnitude, with ITI as the within-subjects factor and order as the between-subjects factor. There was no significant main effect of ITI (fixed vs. random), t-test  indicated that median reaction time (in seconds) did not significantly differ between fixed-ITI (M = 1.001 SD = .199) and random-ITI  blocks, t (29) = .30, p = .769, M (diff) = .008, SE (diff) = .026, 95%CI [−.045–.06], δ = .05. A follow-up Bayesian paired-samples t-test using JASP showed moderate evidence for the null, using a Cauchy prior of .707, BF01 = 4.94, error % = .011.Since reaction times are occasionally used as an indirect measure of task engagement . Only two participants correctly detected that one version of the task had randomised intervals between trials; one of these participants had extensive prior exposure of the original AB task through other research participation. This suggests that most participants were not consciously aware of the temporal difference between AB blocks.This finding does not show evidence that randomising the ITI affects performance in the AB task. A post-hoc Bayesian analysis shows moderate evidence for the null. This result expands upon findings made by Our analysis on order effects in the data was suggested by a reviewer, but since it was a post-hoc analysis we note that it is somewhat underpowered, so these results should be interpreted with caution. However, it is interesting to note that the learning effect from the first to the second session seems somewhat less in the random-first compared to the fixed-first group see . This woThese findings have useful implications for future research, as it provides confirmation that the standard AB task, typically used as a measure of transient attention, has similar levels of interest and difficulty to those tasks which employ a random ITI. Such tasks are typically used as a measure of sustained attention ; The method adopted in the present experiment can be summarised as being a highly pared-back version of the AB task, in conjunction with a brief but randomised ITI, akin to the PVT, though considerably reduced in length. The decision to use the simplified version of the AB task was mainly practical; fewer lags required fewer trials overall, increasing the number of trials possible for each lag and thus giving increased signal-to-noise ratio. However, the rationale for the use of the random interval was more complex.Although most experimental paradigms involving a PVT use longer randomised intervals of 2 to 10 s, this was deemed too long in the current experiment, which instead used a randomised ITI of 0.1 to 3.1 s. Firstly, the inclusion of a longer ITI would drastically alter the length of the two versions of the task, potentially rendering them incomparable. Secondly, it was believed that the use of a longer ITI might appear too obvious in contrast with the fixed ITI, and that this noticeable difference might consciously influence participants’ performance. Indeed, it appeared that the abridged ITI of 0.1 to 3.1 s did successfully evade detection in most cases.It is possible that, during the randomised ITI condition, participants may have performed differently during longer compared to shorter ITIs –there is certainly precedent for this in the literature  had a measurable effect on task performance and engagement in the SNARC effect .Click here for additional data file.10.7717/peerj.8677/supp-2Supplemental Information 2Click here for additional data file.10.7717/peerj.8677/supp-3Supplemental Information 3Click here for additional data file.10.7717/peerj.8677/supp-4Supplemental Information 4This folder contains the MATLAB code necessary to replicate the experiment and analyse the raw data from the paper.Click here for additional data file.10.7717/peerj.8677/supp-5Supplemental Information 5Click here for additional data file.10.7717/peerj.8677/supp-6Supplemental Information 6Each data point represents the proportion of correct responses for T2 given that T1 was identified correctly, from 30 trials.Click here for additional data file.10.7717/peerj.8677/supp-7Supplemental Information 7This analysis in jamovi shows that removing non-responses to T2 from all the analyses produces extremely similar results to those reported in the paper (where non-responses were scored as incorrect).Click here for additional data file.10.7717/peerj.8677/supp-8Supplemental Information 8This file should enable the reader to replicate all the statistical analyses in the paper using any statistical programClick here for additional data file.


For this purpose, we have recently developed a new concept of device based on Electrodes Array for Sampled-Current Voltammetry (EASCV), which is compatible with miniaturization and portability. In this work, to improve the sensitivity of the analytical method, we added a preconcentration step before EASCV analysis, combining sampled-current voltammetry with anodic stripping voltammetry. Lead was chosen as analyte for this probe of concept owing to its high toxicity. The conditions for electrodeposition of lead on gold were optimized by means of under potential deposition. Current intensities 300 times higher than with linear sweep anodic stripping voltammetry were obtained, showing the interest in the method. The value of the sampling time directly affected the sensitivity of the sensor given by the slope of the linear calibration curve. The sensor exhibited a limit of detection of 1.16 mg L Heavy metals such as lead, mercury and cadmium are very toxic even at trace level with European drinking water guidelines set at 10, 1 and 5 μg/L, respectively . To faciElectrochemical studies on this field focused on more sensitive, reproducible and selective systems. The improvement of the detection of heavy metals at trace level requires the optimization of anodic stripping voltammetry. Thus, Krowa-Eisner et al. used Subtractive Anodic stripping voltammetry (SASV) ,19. Thisversus the applied potential. Thus, the renewal of the electrode surface was assured and a fresh solution was always available close to the electrode surface. Since the data acquisition that does not require the use of a potential ramp was simplified, the device can be portable and cost-effective. Our previous studies focused on the interest in the method to mimic dropping mercury electrodes and to circumvent the problem of passivation during analysis.We have previously reported a new analytical method called Electrodes Array for Sampled-Current Voltammetry (EASCV) based on sampled-current voltammetry performed on an electrode array ,23. A poThe aim of this new study is to combine an electrodeposition step with EASCV for heavy metal detection. The use of an electrode array in EASCV allows a coupling of methods that was not possible before with dropping mercury electrodes. Since a new electrode covered by the same amount of metal will be analyzed at each applied potential, high current intensities will be expected with a simplified data processing for an easier adaptation to portable device. Lead detection was chosen in this work as an example of application.3 solution followed by ultra-pure water before use to avoid metal contamination.Lead (II) nitrate or Lead (II) chloride 99% were purchased from Acros and Aldrich, France, respectively. All solutions were prepared with ultra-pure water . All glassware and the electrochemical cell were rinsed with a 10% HNOThe electrodes prepared by photolithography ,24 were 3 69%. After rinsing abundantly with ultra-pure water, the electrode array was washed in several baths of ultra-pure water with slow stirring (50 rpm).After analysis, the electrodes were regenerated in HNOThe electrochemical analyses were performed in a standard three-electrode configuration, with a platinum wire counter electrode, a saturated calomel reference electrode (SCE) and a gold electrode depending on the experiment:2) inserted in a tube of glass (polished before each experimentation)(1) A gold bar ((2) An electrode array  under deaerated conditions in an electrochemical cell adapted to the electrode Concerning the homemade electrode array the electrochemical experiments were performed in a homemade electrochemical cell  22]. Th. Th22]. ® Model potentiostat/galvanostat with a versaSTAT LC Low Current Interface (Princeton Applied Research) and the versaStudio Software.Cyclic voltammetry and chronoamperometry experiments were carried out with a VersaSTAT3 AMETEK−1 sodium chloride or 0.01 mol L−1 KNO3 potassium nitrate as supporting electrolyte according to the nature of the lead salts.The electrochemical analysis was performed in ultra-pure water, containing either 0.1 mol LThe electrochemical signal of lead depends on the nature of the electrode. In this study, gold was chosen as a good alternative to mercury ,13,14 anTwo cathodic peaks beginning at −0.025 V/SCE (1) for UPD1 and −0.275 V/SCE (2) for UPD2 appeared at more positive potentials than the lead equilibrium potential (−0.46 V/SCE). This phenomenon called UPD occurs when a metal is deposited on an electrode of different nature. Thus, the metal is deposited at a potential higher than its equilibrium potential due to a difference in affinity between the metal and the surface of the electrode compared with the metal-metal interaction. The interactions between the metal and the electrode surface are stronger than the metal-metal interactions. Therefore the energy required to reduce the first layer of metal is less important than that required for multilayer formation . The appThe UPD signal in stripping voltammetry analysis offers several advantages . First, Different electrodeposition potentials (Ed) were tested ranging from −0.3 V/SCE to −0.7 V/SCE for the same deposition time (td = 60 s) .The peak current increased when the electrodeposition potential was more negative. A first peak was observed at −0.075 V/SCE. The presence of a shoulder at −0.25 V/SCE for Ed = −0.35 V/SCE showed the appearance of a second peak, which was clearly visible when Ed = −0.5 V/SCE. Finally, a third peak appeared for Ed = −0.7 V/SCE. This peak probably corresponded to the multilayer deposition of lead since it occurred at −0.53 V/SCE under these conditions . It has −7 mol L−1 lead chloride was electrodeposited at a potential of −0.5 V/SCE on a gold electrode of the array.To study the effect of the electrodeposition time, a solution of 5 × 10d.d up to 90 s. A decrease was observed when higher electrodeposition times were used (120 s), probably due to a weak adherence between lead and the gold electrode surface. A time of 90 s was then chosen in further experiments as a good compromise between the analysis time and the current intensity.−9 mol cm−2 [The surface area occupied by lead on a flat gold surface has been estimated to be 1.6 × 10mol cm−2 . For a gThis corresponds to a charge of:This value is 33 times higher than the experimental value, showing that a submonolayer was obtained after an electrodeposition step of 90 s. It confirmed the UPD electrodeposition of lead on the electrode surface.3, followed by extensive rinsing with ultra-pure water.After each analysis, the electrode array was electrochemically oxidized and then washed in concentrated HNO−1 of lead chloride. versus time with and without blank subtraction. The potentials applied to the electrodes of the array were incremented by 0.03 V between −0.25 V/SCE and 0.15 V/SCE. A current peak was observed at short times, which has been previously linked to the response time of the potentiostat [A first test was carried out for a concentration of 15 μmol Lntiostat . HoweverThe variation of the current was higher in the presence of lead compared with the blank, with current values about seven times higher than in the absence of lead a,b.versus time. In The onset potential corresponding to the oxidation of lead was around −0.2 V/SCE, which corresponded to the potential of UPD1 in To show the interest of the method compared to linear sweep anodic stripping voltammetry, an analysis of a lead solution was carried out under the same electrodeposition conditions and with the same electrode surface .As expected, the current intensity of the peak obtained by linear voltammetry was at least 300 times lower than the maximum value obtained by EASCV. The difference was more pronounced if the sampling time was shorter. Indeed, in linear voltammetry, lead deposited on the electrode surface began to be reoxidized from −0.45 V/SCE and when the potential of the peak was reached, a high amount of lead was already oxidized. In EASCV, each electrode was independent of each other and the same initial amount of lead was present on the electrode surface when a new potential was applied. The high current intensities is an advantage of EASCV since the method does not require a Faraday cage and it is also very promising for application with ultramicroelectrodes.2 was performed on the same electrode array.To simplify, the variation of the current as a function of lead concentration was studied by applying three potentials close to the maximum of current observed in θ: 0.005 s and 0.007 s. In This experiment was repeated three times and the average obtained is shown in For highest concentrations of lead, the current decreased when the potential increased, probably due to the higher kinetics of the oxidation reaction. Therefore, the calibration curves were plotted from the current values obtained at 0.07 V/SCE. These calibration curves for the two different sample times are given in With −1 for θ = 0.005 s and 1.41 mg L−1 for 0.007 s. These data clearly demonstrate the influence of the sampling time on the results. First, a shorter sampling time led to higher current intensities and to a higher slope of the calibration curve  was similar to those of EASCV. For comparison, the limit of detections LOD of sensors with gold working electrodes reported in literature are given in −1 in EASCV and 1.02 mg L−1 in linear voltammetry). It underlines that the performances of the sensor should be improved by changing the nature and size of the working electrodes of the array.The sensitivity was significantly lower than for EASCV owing to the lower current range. Owing to this low current range, a Faraday cage was necessary to obtain a good precision of the current values. It is also interesting to note that significantly lower sensitivities were previously reported for anodic stripping voltammetry of lead performed on gold electrode (0.043  and 0.01−1 L ). HoweveThe aim of this article being the effect of coupling EASCV with an electrodeposition step on the current signal, the selectivity of the device was not studied and similar interferents already reported for anodic stripping voltammetry on gold electrode ,13,14,33−1, which was similar to those of anodic stripping voltammetry on a single electrode. This value is high to measure lead in drinking water (10 μg L−1), but it is close to the limit of 0.5 mg L−1 for industrial wastewater according to the decree of 2 February 1998 [This study proposes a new analytical technique for the detection of trace metals by coupling EASCV with anodic stripping voltammetry. As a first advantage, the simplicity of the method would allow analyses on field with a portable system. Since a high electrochemical signal was expected with this method, under potential deposition of lead on the gold electrode array was used to simplify the electrochemical signal. Interestingly, the comparison of this technique with linear voltammetry showed that the maximum current intensity was 300 times higher for a same concentration. It results in a significantly higher sensitivity of the sensor given by the slope of the linear calibration curve. This result is very promising since it underlines the interest of coupling EASCV with a preconcentration step. The application of this first sensor for lead detection led to a limit of quantification of 1.16 mg Lary 1998 . An impr


Instead of having affiliations The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.


Instead of “The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.


An incomplete disclaimer was used.In the original article, there was an error. Disclaimer:A correction has been made to Disclaimer: The findings and conclusions in this publication are those of the author(s) and should not be construed to represent any official USDA or U.S. Government determination or policy. Mention of any trade name, proprietary product or specific equipment does not constitute a guarantee or warranty by USDA-ARS and does not imply its approval to the exclusion of other products that may also be suitable. The USDA-ARS is an equal opportunity and affirmative action employer and all agency services are available without discrimination.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.


Background and Objectives: Metabolic syndrome (MetS) is a multiple risk factor for atherosclerosis, cardiovascular disease, type 2 diabetes and strokes. One-third of middle-age women are at risk of MetS, which predisposes them to type 2 diabetes and cardiovascular disease. Changes in the regulation of anti-inflammatory cytokines―which play an important role in pathologic processes―may contribute to inflammatory disorders. Cytokine polymorphisms are known to have an impact on gene expression. The purpose of this study was to search for the relationship between the IFNG polymorphisms and the levels of proinflammatory cytokines. Materials and Methods: This study, conducted in West Pomeranian Voivodeship, Poland, involved 416 women. Of these women, 33.6% of them had primary education, 44.8% lived in cities with a population of over 100,000, and 82.7% were married. Of the participants, 28.4% met the criteria for MetS. The study involved: interview performed to collect sociodemographic and medical data, anthropometric measurements, as well as venous blood collection for biochemical analysis, genetic testing and the measurement of inflammatory markers. Results: The link between the IFNG (rs2430561) polymorphism and serum PIC (proinflammatory cytokines) levels was tested with regard to MetS. In the MetS+ subgroup, the T/T and A/T genotypes of the IFNG gene were accompanied by higher IL-6 levels than in the MetS− subgroup. Conclusion: Our study has not confirmed a direct link between the IFNG polymorphisms and the levels of inflammatory biomarkers. Nevertheless, the T/T and A/T genotypes of the IFNG gene may predispose to elevated IL-6 levels. Metabolic syndrome (MetS) is a multiple risk factor for atherosclerosis, cardiovascular disease, type 2 diabetes and strokes. It is a worldwide health problem; whose incidence rate ranges from 13.8% to over 60% in different populations . MetS coAs a result of reduced sex hormone secretion in the perimenopausal period―a natural consequence of gonadal dysfunction and aging― many women develop chronic diseases from the sixth decade onwards, which affects their quality of life. As defined by the stages of reproductive aging workshop (STRAW) criteria, perimenopause is the time between the first major changes in the length of the menstrual cycle  and the period of 12 consecutive months without menstruation . PotentiOne-third of middle-age women are at risk of developing MetS, which predisposes them to type 2 diabetes and cardiovascular disease (CVD) ,15,16.Obesity, insulin resistance and type 2 diabetes entail chronic inflammation, resulting from abnormal levels of cytokines, acute phase reactants and other inflammatory signaling markers . MetS isIFNG +874 A/T single nucleotide polymorphism (SNP) located at the position +874 of intron 1 can affect the secretion of IFNγ. The IFNγ production is determined by the polymorphic IFNG (rs2430561) gene, which includes the T allele, contributing to high IFNγ production and the A allele responsible for low IFNγ production. Average levels of IFNγ cytokines are higher in healthy carriers of the T allele than in those with the A allele. It has been clearly demonstrated that the T-to-A polymorphism has a direct effect on the level of IFNγ production [Changes in the regulation of anti-inflammatory cytokines―which play an important role in pathologic processes―may contribute to inflammatory disorders. Cytokine polymorphisms are known to have an impact on gene expression. Interferon gamma (IFNγ) is the main proinflammatory cytokine. Studies show that the oduction .IFNG polymorphisms and the levels of proinflammatory cytokines  in women aged 45–60 years.The purpose of this study was to analyze the relationship between the This study, conducted in West Pomeranian Voivodeship (Poland), involved 416 women, whose mean age was 53.5 years. The inclusion criteria were female sex, the age of 45–60 years, using MHT and no history of inflammatory, psychiatric or neoplastic diseases. All participants were non-smokers, drank less than 20 g of pure alcohol per day or drank occasionally no more than 40 g of pure alcohol and abstained from alcohol at least two days per week. Of the participants, 33.6% had primary education, 27.7% had tertiary education, 44.8% lived in cities with a population of over 100,000, almost 40% lived in rural areas, 82.7% of the women were married, 28.4% met the criteria for MetS. Each participant gave informed written consent to take part in this study.The study was carried out in accordance with ethical standards and the Declaration of Helsinki. The protocol was approved by the bioethical committee of the Pomeranian Medical University, Szczecin, Poland on 19 June 2017 .IFNG (rs2430561) polymorphism. The biologic material for DNA analysis was stored and transported in accordance with the quality management system of the Genetic Laboratory, Clinic and Department of Psychiatry .The study involved: interview performed to collect sociodemographic and medical data ; anthropometric measurements ; venous blood collection for biochemical analysis, genetic testing and the measurement of inflammatory markers in accordance with the procedures for sampling and transport of the biologic material. Blood was taken between 7.00 and 9.30 a.m. after overnight fasting and ten-minute rest in a sitting position. Blood was collected to two Vacutainer tubes : a tube containing 1-g/L dipotassium (K2) ethylenediaminetetraacetic acid (EDTA) and a tube for biochemical serum analysis (7 mL). The levels of triglycerides, HDL and glycemia after overnight fasting were determined. Next, DNA was isolated for genetic analysis of the Additionally, the levels of inflammatory markers  were determined.n = 118) comprising respondents who met the criteria for MetS based on the International diabetes federation (IDF) classification [n = 298)—including those who did not meet the above-mentioned criteria. The respondents were assigned to the MetS+ subgroup if they had at least three out of the five components of MetS: waist size ≥80 cm; glycemia on an empty overnight fasting ≥100 mg/dL (5.6 mmol/L) or related pharmacotherapy; the levels of triglycerides ≥150 mg/dL (1.7 mmol/L) or related pharmacotherapy; HDL cholesterol ≤50 mg/dL (1.3 mmol/L) or related pharmacotherapy; systolic blood pressure ≥130 and/or diastolic blood pressure ≥85 mmHg or related pharmacotherapy.Based on the results of laboratory and anthropometric measurements, the women were qualified to two subgroups: the MetS+ subgroup (fication  and the p < 0.05. All tests were two-tailed. The two groups (MetS+ and MetS−) were compared using the Student’s t-test in the case of normal data distribution and the Mann–Whitney U test in the case of skewed data distribution. To assess the link between MetS and particular genotypes, four inheritance models were tested with the Bayesian information criterion (BIC) in order to select the best one. An effect of particular variables and alleles on the development of MetS was calculated using odds ratio (OR) with 95% confidence interval (CI).Statistical analysis was performed using Statistica 13.0 PL  and R software (R Core Team (2013). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria). Statistical significance was set as The results presented in this manuscript are a part of the larger project performed by a team of researchers.p < 0.001) between waist size, the levels of glycemia after overnight fasting, triglycerides and cholesterol, systolic and diastolic blood pressure and MetS. A statistically significant relationship (p < 0.05) was also noticed between Mets and the level of IL-6. MetS was not statistically significantly related to the levels of IL-1α, IL-1β, TNFα and IFNγ  polymorphism with regard to MetS. The T/T genotype was found in 45% of the participants, the A/T genotype in 39% and the A/A genotype in 16%. The T allele was more common (64%) than the A allele (36%). There were no significant differences in the distribution of the genotypes and alleles between the participants who met the criteria for MetS and those who did not. We tested the link between the IFNG (rs2430561) polymorphism and serum PIC levels with regard to MetS. In the MetS+ subgroup, the T/T and A/T genotypes of the IFNG gene were accompanied by higher IL-6 levels than in the MetS− subgroup.We analyzed the distribution of the genotypes and alleles of the IFNG gene in any of the MetS subgroups  in older women with and without MetS, found that those with MetS had higher levels of cytokines and risk factors for cardiovascular disease  [ificant) . Other aificant) . In a stificant) .IFNG +874 T polymorphism has an effect on the expression of this gene. The A/A genotype was accompanied by lower levels of IFNγ than the T/T genotype and the heterozygous T/A genotype entailed average IFNγ levels [IFNG +874 T/A polymorphism was linked to lower levels of total cholesterol and triglycerides [In our study, MetS was significantly related to the levels of IL-6, but not to the levels of IL-1α, IL-1β, TNFα and IFNγ. The results show that the γ levels . IFNγ taγ levels . Polymorγ levels . It was ycerides .IFNG +874 polymorphism on the levels of selected inflammatory biomarkers in patients with and without MetS. Respondents having no history of inflammatory, psychiatric or cancerous diseases and not using MHT were qualified to participate in the study. All participants were non-smokers and did not drink excess alcohol. The results did not reveal significant differences in the distribution of genotypes and alleles between the Mets+ and the Mets− subgroups. However, in the MetS+ subgroup, the T/T and A/T genotypes of the IFNG gene were accompanied by higher IL-6 levels than in the MetS− subgroup. There were no significant differences in the levels of other biomarkers .We analyzed the influence of the IFNG gene on the level of IL-6. The study by Kim et al. demonstrated that the risk of cardiovascular disease, estimated using the Framingham risk score (FRS ≥ 10%), was associated with the socioeconomic status of the respondents more than with Mets [The study presented here has some limitations. Due to the substantial influence of sociodemographic factors and health behaviors on the respondents’ health status, we are not able to clearly determine the influence of the T/T and A/T genotypes of the ith Mets . Anotherith Mets . It is aith Mets . Furtherith Mets .IFNG polymorphisms and the levels of the inflammatory biomarkers in perimenopausal women analyzed with regard to MetS. However, the presence of the T/T and A/T genotypes of the IFNG gene may predispose to elevated IL-6 levels.Our study has not confirmed a direct link between the


Mugilogobius myxodermus is an endemic freshwater fish in china. Herein, we sequenced and assembled the first complete mitogenome of M. myxodermus. The total length of the mitogenome is 16,495 bp, containing 13 protein-coding genes, two ribosomal RNAs (rRNA), two non-coding regions (control region and origin of light-strand replication), and 22 transfer RNAs (tRNA). The overall base composition of M. myxodermus is 26.76% T, 28.97% C, 27.81% A, and 16.45% G, respectively. A phylogenetic tree showed that M. myxodermus clustered closest to M. abei. The complete mitogenome of M. myxodermus provides a resource for studies on biogeography and evolution of this gobiid fish.The  Mugilogobius myxodermus . The specimens of M. myxodermus were collected from the Lake Tai at Wuxi, China  and identified by morphology (Larson http://www.lasdr.cn/pages/resdata_dataview.jsp?id=10069100). Total genomic DNA was extracted from the tail fin with the HiPure Tissue DNA Mini Kit . The versatile primers were designed on the basis of the mitogenome sequence of Mugilogobius abei (GenBank Accession No. NC_023353) , two non-coding regions (control region and origin of light-strand replication), and 22 transfer RNAs (tRNA). Except for the eight tRNAs  and ND6 genes encoded on the L-strand, all other mitochondrial genes are encoded on the H-strand. The overall nucleotide composition of M. myxodermus is 26.76% T, 28.97% C, 27.81% A, and 16.45% G, respectively. All 13 protein-coding genes start with ATG except COXI which starts with GTG. Most of the protein-coding genes (10 of 13 genes) end with TAA, TAG, and AGG, the other three genes  have T or TA incomplete stop condon, which are very typical in many other gobiid fishes (Chen and Wen M. myxodermus contains two non-coding regions: the control region (D-loop) is 850 bp in length, which is located between ProtRNA and PhetRNA genes; another small non-coding region, which is a 36 bp fragment and the origin of light-strand replication (OL), is located between the AsntRNAand CystRNAgenes. The gene content, structure, and arrangement of mitogenome in M. myxodermus are similar to those observed in other fishes  were mined from GenBank. A maximum likelihood phylogenetic analysis was conducted using the software Mega 6 (M. myxodermus was phylogenetically placed near the M. abei and M. chulae, with a very short branch length. The complete mitochondrial genome of M. myxodermus will be useful for further evolution and biogeography studies of gobiid fishes.All available full mitogenome sequences of subfamily Gobionellinae and two non-percomorph outgroups (e Mega 6  (Tamura e Mega 6 . M. myxo