Role of cold shock proteins B and D in Aeromonas salmonicida subsp. salmonicida physiology and virulence in lumpfish (Cyclopterus lumpus).

Cold shock proteins (Csp) are pivotal nucleic acid binding proteins known for their crucial roles in the physiology and virulence of various bacterial pathogens affecting plant, insect, and mammalian hosts. However, their significance in bacterial pathogens of teleost fish remains unexplored. Aeromonas salmonicida subsp. salmonicida (hereafter A. salmonicida) is a psychrotrophic pathogen and the causative agent of furunculosis in marine and freshwater fish. Four csp genes (cspB, cspD, cspA, and cspC) have been identified in the genome of A. salmonicida J223 (wild type). Here, we evaluated the role of DNA binding proteins, CspB and CspD, in A. salmonicida physiology and virulence in lumpfish (Cyclopterus lumpus). A. salmonicida ΔcspB, ΔcspD, and the double ΔcspBΔcspD mutants were constructed and characterized. A. salmonicida ΔcspB and ΔcspBΔcspD mutants showed a faster growth at 28°C, and reduced virulence in lumpfish. A. salmonicida ΔcspD showed a slower growth at 28°C, biofilm formation, lower survival in low temperatures and freezing conditions (-20°C, 0°C, and 4°C), deficient in lipopolysaccharide synthesis, and low virulence in lumpfish. Additionally, ΔcspBΔcspD mutants showed less survival in the presence of bile compared to the wild type. Transcriptome analysis revealed that 200, 37, and 921 genes were differentially expressed in ΔcspB, ΔcspD, and ΔcspBΔcspD, respectively. In ΔcspB and ΔcspBΔcspD virulence genes in the chromosome and virulence plasmid were downregulated. Our analysis indicates that CspB and CspD mostly act as a transcriptional activator, influencing cell division (e.g., treB), virulence factors (e.g., aexT), and ultimately virulence.

Socioeconomic and geographical inequalities in delivery by cesarean section among women in Bangladesh, 2004-2017.

BACKGROUND: There is a dearth of evidence on the trends and inequalities in utilizing cesarean section (CS) among women in Bangladesh. Hence, this study aimed to estimate the socioeconomic and geographical inequalities in delivery by CS among Bangladeshi women from 2004 to 2017. METHODS: Data from Bangladesh Demographic and Health Survey 2004, 2007, 2011, 2014, and 2017 were analyzed using the WHO's Health Equity Assessment Toolkit (HEAT) software. Inequalities were measured using four summary measures: Difference (D), Population Attributable Risk (PAR), Population Attributable Fraction (PAF), and Ratio (R). Socioeconomic inequalities were assessed using two equity dimensions: household wealth status, and level of education, while geographical disparities were measured using two equity dimensions: place of residence, and sub-national regions. For each measure, point estimates and their 95% confidence intervals were reported. RESULTS: An increasing trend in the prevalence (weighted) of CS in Bangladesh use was found from 4.50% in 2004 to 32.73% in 2017 We found significant socioeconomic inequalities in CS in every survey point, with a higher concentration of CS among the rich (in 2017, PAR = 28.57; 95% CI: 26.69-30.46) indicating a pro-rich inequality, and higher educated (in 2017, PAF = 23.97; 95% CI: 12.26-35.68) sub-groups. We also identified significant geographical disparities in CS with a higher concentration of CS among people from urban areas (in 2017, PAR = 10.99; 95% CI: 10.19-11.79), and a coastal region (Khulna division) (in 2017, PAF: 30.48 (95% CI: 18.66-42.30). CONCLUSION: We observed both socioeconomic and geographical inequalities in CS exist in Bangladesh, though the trends of these inequalities were curved over time. Thus, it is important to comprehend these pro-rich and geographical inequalities better and implement appropriate interventions and policies to alleviate them.

Validity and reliability of an Arabic-language version of the postpartum specific anxiety scale research short-form in Jordan.

OBJECTIVE: The English-language Postpartum Specific Anxiety Scale (PSAS) is a valid, reliable measure for postpartum anxiety (PPA), but its 51-item length is a limitation. Consequently, the PSAS Working Group developed the PSAS Research Short-Form (PSAS-RSF), a statistically robust 16-item tool that effectively assesses PPA. This study aimed to assess and validate the reliability of an Arabic-language version of the PSAS-RSF in Jordan (PSAS-JO-RSF). METHODS: Using a cross-sectional methodological design, a sample of Arabic-speaking mothers (N = 391) with infants aged up to 6 months were recruited via convenience sampling from a prominent tertiary hospital in northern Jordan. Factor analysis, composite reliability (CR), average variance extracted (AVE), McDonald's ω, and inter-item correlation measures were all examined. RESULTS: Explanatory factor analysis revealed a four-factor model consistent with the English-language version of the PSAS-RSF, explaining a cumulative variance of 61.5%. Confirmatory factor analysis confirmed the good fit of the PSAS-JO-RSF (χ2/df = 1.48, CFI = 0.974, TLI = 0.968, RMSEA = 0.039, SRMR = 0.019, p < 0.001). The four factors demonstrated acceptable to good reliability, with McDonald's ω ranging from 0.778 to 0.805, with 0.702 for the overall scale. The CR and AVE results supported the validity and reliability of the PSAS-JO-RSF. CONCLUSION: This study establishes an Arabic-language version of the PSAS-JO-RSF as a valid and reliable scale for screening postpartum anxieties in Jordan.

Assessing the Gap Between Women's Expectations and Perceptions of the Quality of Intrapartum Care in Jordan: A Two-Stage Study Using the SERVQUAL Model.

INTRODUCTION: Although Jordan has made significant progress toward expanding the utilization of facility-based intrapartum care, prior research highlights that poor service quality is still persistent. This study aimed to identify quality gaps between women's expectations and perceptions of the actual intrapartum care received, while exploring the contributing factors. METHODS: Utilizing a pre-post design, quality gaps in intrapartum care were assessed among 959 women pre- and postchildbirth at a prominent tertiary hospital in northern Jordan. Data were gathered using the SERVQUAL scale, measuring service quality across reliability, responsiveness, tangibles, assurance, and empathy dimensions. RESULTS: The overall mean gap score between women's expectations and perceptions of the quality of intrapartum care was -0.60 (±0.56). The lowest and highest mean gap scores were found to be related to tangibles and assurance dimensions, -0.24 (±0.39) and -0.88 (±0.35), respectively. Significant negative quality gaps were identified in the dimensions of assurance, empathy, and responsiveness, as well as overall service quality (p < 0.001). The MLR analyses highlighted education (ß = 0.61), mode of birth (ß = -0.60), admission timing (ß = -0.41), continuity of midwifery care (ß = -0.43), physician's gender (ß = -0.62), active labour duration (ß = 0.37), and pain management (ß = -0.33) to be the key determinants of the overall quality gap in intrapartum care. CONCLUSION: Our findings underscore the importance of fostering a labour environment that prioritizes enhancing caregivers' empathetic, reassuring, and responsive skills to minimize service quality gaps and enhance the overall childbirth experience for women in Jordan. PATIENT OR PUBLIC CONTRIBUTION: This paper is a collaborative effort involving women with lived experiences of childbirth, midwives, and obstetrics and gynaecologist physicians. The original idea, conceptualization, data generation, and coproduction, including manuscript editing, were shaped by the valuable contributions of stakeholders with unique perspectives on intrapartum care in Jordan.

Inactivated Aeromonas salmonicida impairs adaptive immunity in lumpfish (Cyclopterus lumpus).

Aeromonas salmonicida, a widely distributed aquatic pathogen causing furunculosis in fish, exhibits varied virulence, posing challenges in infectious disease and immunity studies, notably in vaccine efficacy assessment. Lumpfish (Cyclopterus lumpus) has become a valuable model for marine pathogenesis studies. This study evaluated several antigen preparations against A. salmonicida J223, a hypervirulent strain of teleost fish, including lumpfish. The potential immune protective effect of A. salmonicida bacterins in the presence and absence of the A-layer and extracellular products was tested in lumpfish. Also, we evaluated the impact of A. salmonicida outer membrane proteins (OMPs) and iron-regulated outer membrane proteins (IROMPs) on lumpfish immunity. The immunized lumpfish were intraperitoneally (i.p.) challenged with 104 A. salmonicida cells/dose at 8 weeks-post immunization (wpi). Immunized and non-immunized fish died within 2 weeks post-challenge. Our analyses showed that immunization with A. salmonicida J223 bacterins and antigen preparations did not increase IgM titres. In addition, adaptive immunity biomarker genes (e.g., igm, mhc-ii and cd4) were down-regulated. These findings suggest that A. salmonicida J223 antigen preparations hinder lumpfish immunity. Notably, many fish vaccines are bacterin-based, often lacking efficacy evaluation. This study offers crucial insights for finfish vaccine approval and regulations.

Assessing health-related quality of life among cancer survivors during systemic and radiation therapy in Bangladesh: a cancer-specific exploration.

BACKGROUND: Evaluating the effects of cancer diagnosis and treatment on a patient's overall well-being is crucial and health-related quality of life (HRQoL) is a reliable metric for assessing this impact. Little is known about HRQoL among cancer survivors across various stages and treatments. The study examined individual and clinical factors influencing HRQoL among cancer survivors. METHODS: A cross-sectional study was conducted in two specialised cancer care hospitals in Dhaka, Bangladesh. Cancer-diagnosed adults receiving treatment at selected hospitals from January to May 2022 were enrolled. The 5-level EuroQol-5 Dimensions version (EQ-5D-5L) instrument was used to collect HRQoL data. HRQoL scores were derived using UK value sets. The investigation used a multivariable Tobit regression model to determine the association between independent variables and HRQoL scores. RESULTS: A total of 607 adult patients were enrolled, with 55% being females and 66% aged 36 to 64 years. Reported health problems in five EQ-5D domains include mobility (11%), self-care (11%), usual daily activities (19%), pain/discomfort (21%), and anxiety/depression (46%). Patients with throat, brain, lung, blood, and liver cancer had lower utility scores. Advanced-stage cancer survivors had lower utility scores (ß = -49 units, 95% codfidence interval [CI]: -0.75 to -0.22) compared to early-stage survivors. Physically inactive survivors had lower utility scores by 0.41 units (95% CI: -0.51 to -0.30) compared to their counterparts. Private hospital patients had higher utility scores, whereas patients belonged to poor socioeconomic groups scored worse than wealthier ones. CONCLUSIONS: This study highlights the impact of clinical and individual characteristics on HRQoL among cancer survivors. These findings advocate for an enhanced Bangladeshi cancer patient care model through timely interventions or programs, early detection or diagnosis, tailored treatments, and the promotion of physical activity to bolster HRQoL outcomes.

Prevalence and determinants of physical violence against doctors in Bangladeshi tertiary care hospitals.

BACKGROUND: The increasing physical violence against doctors in the health sector has become an alarming global problem and a key concern for the health system in Bangladesh. This study aimed to determine the prevalence and associated factors of physical violence against doctors in Bangladeshi tertiary care hospitals. METHODS: A cross-sectional survey was performed among 406 doctors working in tertiary care hospitals. Data were collected using a self-administered questionnaire and the binary logistic regression model was employed for predicting physical violence against doctors. RESULTS: Of the participants, 50 (12.3%) doctors reported being exposed to physical violence in 12 months prior to the survey. According to logistic regression analysis, aged less than 30 years or younger, male and never-married doctors were prone to physical violence. Similarly, doctors from public hospitals and those worked in emergency departments were at higher risk of physical violence. More than 70% of victims reported that patients' relatives were the main perpetrators. Two-thirds of the victims referred to violence in the hospitals as a grave concern. CONCLUSIONS: Physical violence against doctors is relatively common in the emergency departments and public hospitals in Bangladesh. This study found that male and younger doctors were at high risk of exposing physical violence. To prevent hospital violence, authorities must develop human resources, bolster patient protocol and offer physician training.

Association between inter-pregnancy interval and risk of adverse birth outcomes in subsequent pregnancy: A retrospective study from Jordan.

BACKGROUND: Adverse birth outcomes, such as preterm birth and low birth weight (LBW), are leading causes of neonatal morbidity and mortality. In this study, we aimed to estimate the association between inter-pregnancy interval (IPI) and the risks of adverse birth outcomes in a subsequent pregnancy. METHODS: We conducted a retrospective analysis involving 630 mothers who delivered a singleton live infant at a leading tertiary hospital in northern Jordan from March to August 2021. Outcome variables were preterm birth (<37 weeks of gestation) and LBW (<2.5 kg). Using multivariable logistic regression, the association between IPI and these two adverse birth outcomes was investigated. RESULTS: The rates of preterm birth and LBW were 12.4% and 16.8%, respectively. Compared with an optimal IPI (24-36 months), a short IPI (<24 months) was positively associated with preterm birth (aOR: 4.09; 95% CI: 1.48-6.55) and LBW (aOR: 3.58; 95% CI: 1.57-5.15). Infants conceived after a long IPI (≥ 60 months) had increased odds of preterm birth (aOR: 3.78; 95% CI: 1.12-5.78) and LBW (aOR: 2.65; 95% CI: 1.67-4.03). Preterm delivery was also significantly associated with the mother's age (aOR: 1.10; 95% CI: 1.04-1.17), history of multiple cesarean births (aOR: 2.67; 95% CI: 1.14-4.29), prolonged rupture of membranes (aOR: 2.46; 95% CI: 1.10-5.52), and perinatal death (aOR: 3.42; 95% CI: 1.10-5.49). A mother's history of prior LBW (aOR: 4.39; 95% CI: 1.08-6.80), hypertensive disorders (aOR: 1.95; 95% CI: 1.03-3.89), and multiple cesarean births (aOR: 4.35; 95% CI: 2.10-6.99) was associated with LBW. CONCLUSIONS: Both short and long IPIs were related to preterm delivery and LBW. Optimal birth spacing is recommended to improve birth outcomes and must be considered when designing effective family planning programs.

Factors influencing and changes in childhood vaccination coverage over time in Bangladesh: a multilevel mixed-effects analysis.

INTRODUCTION: This study aimed to investigate the associated factors and changes in childhood vaccination coverage over time in Bangladesh. METHODS: Bangladesh's Demographic and Health Surveys from 2011, 2014, and 2017-18 provided data for this study on vaccination coverage among children aged 12 to 35 months. For three survey periods, multilevel binary logistic regression models were employed. RESULTS: The overall prevalence (weighted) of full vaccination among children aged 12-35 months were 86.17% in 2011, 85.13% in 2014, and 89.23% in 2017-18. Children from families with high wealth index, mothers with higher education, and over the age of 24 and who sought at least four ANC visits, as well as children from urban areas were more likely to receive full vaccination. Rangpur division had the highest change rate of vaccination coverage from 2011 to 2014 (2.26%), whereas Sylhet division had the highest change rate from 2014 to 2017-18 (34.34%). CONCLUSION: To improve immunization coverage for Bangladeshi children, policymakers must integrate vaccine programs, paying special attention to mothers without at least a high school education and families with low wealth index. Increased antenatal care visits may also aid in increasing the immunization coverage of their children.

A formative research to explore the programmatic approach of vaccinating the Rohingya refugees and host communities against COVID-19 infection in Bangladesh.

BACKGROUND: The vaccination of the Rohingya refugees and host communities against COVID-19 in Cox's Bazar started in August 2021. Government authorities and Non-Government Organisation partners implemented a project around the initial period of vaccination to improve awareness and access to target beneficiaries. We conducted formative research to understand the programmatic approach of this project and identify potential challenges and community perceptions regarding immunisation against COVID-19. METHODOLOGY: This was formative research in which we used a qualitative method of data collection. Purposively chosen 12 key-informant interviews and conveniently chosen 20 in-depth interviews were conducted using semi-structured interview guidelines from August to September 2022 in the Rohingya camp and host communities of Cox's Bazar District, Bangladesh. Ethical approval was obtained from the North South University Institutional Review Board, and written informed consent was obtained from all the participants. We used a thematic analysis approach to analyse the data. RESULTS: The project neither provided any promotional or tailored messages regarding the COVID-19 vaccine nor conducted any vaccine hesitancy surveys before implementing the project. The project did not provide any storage facilities for the vaccines' cold chain management but provided transport support to carry the vaccines from the district to the sub-district level. Community leaders were included in the decision-making process during local-level planning of the vaccination programme. The project supported the reporting of any adverse effects following immunisation from community members to the government health authorities. Vaccine hesitancy among participants was high in the early stages, but mass campaigns and vaccination of frontline health workers increased their acceptance. The major challenges reported by the informants were low budget and lower salaries of field staff, stacking of the registration process at the beginning, reluctance of participants, inadequate transportation and manpower, and inadequate baby feeding corners at vaccination centers. CONCLUSION: The findings from our study will help policymakers from the Government, the UN, and other humanitarian agencies to adapt and better address the issue of vaccine acceptance and strengthen the vaccination programme.

Pseudouridylation defect due to DKC1 and NOP10 mutations causes nephrotic syndrome with cataracts, hearing impairment, and enterocolitis.

RNA modifications play a fundamental role in cellular function. Pseudouridylation, the most abundant RNA modification, is catalyzed by the H/ACA small ribonucleoprotein (snoRNP) complex that shares four core proteins, dyskerin (DKC1), NOP10, NHP2, and GAR1. Mutations in DKC1, NOP10, or NHP2 cause dyskeratosis congenita (DC), a disorder characterized by telomere attrition. Here, we report a phenotype comprising nephrotic syndrome, cataracts, sensorineural deafness, enterocolitis, and early lethality in two pedigrees: males with DKC1 p.Glu206Lys and two children with homozygous NOP10 p.Thr16Met. Females with heterozygous DKC1 p.Glu206Lys developed cataracts and sensorineural deafness, but nephrotic syndrome in only one case of skewed X-inactivation. We found telomere attrition in both pedigrees, but no mucocutaneous abnormalities suggestive of DC. Both mutations fall at the dyskerin-NOP10 binding interface in a region distinct from those implicated in DC, impair the dyskerin-NOP10 interaction, and disrupt the catalytic pseudouridylation site. Accordingly, we found reduced pseudouridine levels in the ribosomal RNA (rRNA) of the patients. Zebrafish dkc1 mutants recapitulate the human phenotype and show reduced 18S pseudouridylation, ribosomal dysregulation, and a cell-cycle defect in the absence of telomere attrition. We therefore propose that this human disorder is the consequence of defective snoRNP pseudouridylation and ribosomal dysfunction.

Workplace violence against Bangladeshi registered nurses: A survey following a year of the COVID-19 pandemic.

AIMS: To investigate the prevalence of workplace violence and its associated factors among Bangladeshi registered nurses. BACKGROUND: Workplace violence is prevalent among nurses, particularly in developing countries. However, the issue has never been examined in Bangladeshi nurses. METHODS: Between February 26 and July 10, 2021, this cross-sectional survey involving 1264 registered nurses was conducted. Workplace violence was determined by the Workplace Violence Scale (WVS). A multivariable logistic regression model was fitted to find the factors associated with workplace violence. This study complies with the EQUATOR (STROBE) checklist. RESULTS: Of the 1264 nurses, 885 (70%) nurses reported being exposed to workplace violence in the previous year. Three hundred twenty-four (324; 25.6%) nurses reported physical violence, whereas 902 (71.4%) nurses reported nonphysical violence. According to the multivariable logistic regression model, male nurses, nurses in the Sylhet division, emergency department nurses, nurses working extended hours, and non trained nurses to tackle workplace violence were prone to physical violence. Furthermore, public hospital nurses and non trained nurses to tackle workplace violence were more likely to be exposed to nonphysical violence. Nurses who had not been exposed to workplace violence were satisfied with their current job, but those who had been exposed to workplace violence were dissatisfied and intended to leave their current job. CONCLUSIONS AND IMPLICATIONS FOR NURSING AND HEALTH POLICY: High prevalence of workplace violence underscores nurses' current working conditions, which are particularly poor in public hospitals and emergency departments. Moreover, the COVID-19 pandemic put unprecedented pressure on the whole healthcare system and caused various difficulties for healthcare workers. To develop a zero-violence practice environment, health authorities should implement policy-level interventions. Healthcare staff should be guided to deal more successfully with patients and coworkers to create a positive working environment.

Prevalence and determinants of contraceptive method use among Bangladeshi women of reproductive age: a multilevel multinomial analysis.

BACKGROUND: Much scholarly debate has centered on Bangladesh's family planning program (FPP) in lowering the country's fertility rate. This study aimed to investigate the prevalence of using modern and traditional contraceptive methods and to determine the factors that explain the contraceptive methods use. METHODS: The study used data from the 2017-18 Bangladesh Demographic and Health Survey (BDHS), which included 11,452 (weighted) women aged 15-49 years in the analysis. Multilevel multinomial logistic regression was used to identify the factors associated with the contraceptive method use. RESULTS: The prevalence of using modern contraceptive methods was 72.16%, while 14.58% of women used traditional methods in Bangladesh. In comparison to women in the 15-24 years age group, older women (35-49 years) were more unwilling to use modern contraceptive methods (RRR: 0.28, 95% CI: 0.21-0.37). Women who had at least a living child were more likely to use both traditional and modern contraceptive methods (RRR: 4.37, 95% CI: 3.12-6.11). Similarly, given birth in the previous 5 years influenced women 2.41 times more to use modern method compared to those who had not given birth (RRR: 2.41, 95% CI: 1.65-3.52). Husbands'/partners' decision for using/not using contraception were positively associated with the use of both traditional (RRR: 4.49, 95% CI: 3.04-6.63) and modern methods (RRR: 3.01, 95% CI: 2.15-4.17) rather than using no method. This study suggests rural participants were 21% less likely to utilize modern methods than urban participants (RRR: 0.79, 95% CI: 0.67-0.94). CONCLUSION: Bangladesh remains a focus for contraceptive use, as it is one of the most populous countries in South Asia. To lower the fertility rate, policymakers may design interventions to improve awareness especially targeting uneducated, and rural reproductive women in Bangladesh. The study also highlights the importance of male partners' decision-making regarding women's contraceptive use.

HIV infection and multidrug resistant tuberculosis: a systematic review and meta-analysis.

BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) in HIV infected individuals is a serious threat to global efforts to combat tuberculosis. Inconsistent findings on the association between HIV infection and MDR-TB were present in many studies. We aimed to review existing data on the relationship between HIV infection and MDR-TB systematically to assess the contribution of HIV on MDR-TB worldwide. We also investigated the patterns of MDR-TB by age, country-wise income, study designs, and global regions. METHODS: We utilized PubMed, Google Scholar, and ScienceDirect databases to select eligible studies for meta-analysis that were published between January 12,010, and July 30, 2020. The random-effects model was used to obtain the pooled odds ratio of the crude association between HIV and MDR-TB with a 95% confidence interval. We investigated the potential publication-bias by checking funnel plot asymmetry and using the Egger's test. Moreover, we assessed the heterogeneity using the I2 statistic. Sensitivity analysis was performed based on sample size and adjustment factors. The protocol was registered with PROSPERO-CRD42019132752. RESULTS: We identified 1603 studies through a database search, and after subsequent eliminations we selected 54 studies including 430,534 TB patients. The pooled odds of MDR-TB was 1.42 times higher in HIV-positive patients than HIV-negative patients (OR=1.42,CI=1.17-1.71, I2=75.8%). Subgroup analysis revealed that the estimated pooled odds for South-East Asian countries was 1.86, which is the highest in WHO regions (OR=1.86,CI=1.30-2.67, I2=0.00%), followed by Europe and Africa. The effect estimate was found to be higher for primary MDR-TB (OR=2.76,CI=1.70-4.46, I2=0.00%). There was also a trend towards increased odds of MDR-TB for HIV patients older than 40 years (OR=1.56,CI=1.17-2.06). The association was found to be significant in high-burden TB countries (OR=1.75, CI=1.39-2.19) and in high-income countries (OR=1.55, CI=1.06-2.27). CONCLUSION: Such findings indicate that HIV infection raises the risk of MDR-TB, and after contrasting it with the results of the earlier pooled study, it appeared to be an upward risk trend. Moreover, we found that the risk is the highest in the South-East Asian region. A balanced allocation of resources is needed to halt both primary and secondary MDR-TB, particularly in HIV infected people with 40 years of age and older.

Aeromonas salmonicida infection kinetics and protective immune response to vaccination in sablefish (Anoplopoma fimbria).

Effective vaccine programs against Aeromonas salmonicida have been identified as a high priority area for the sablefish (Anoplopoma fimbria) aquaculture. In this study, we established an A. salmonicida infection model in sablefish to evaluate the efficacy of commercial vaccines and an autogenous vaccine preparation. Groups of 40 fish were intraperitoneally (ip) injected with different doses of A. salmonicida J410 isolated from infected sablefish to calculate the median lethal dose (LD50). Samples of blood, head kidney, spleen, brain, and liver were also collected at different time points to determine the infection kinetics. The LD50 was estimated as ~3 × 105 CFU/dose. To evaluate the immune protection provided by an autogenous vaccine and two commercial vaccines in a common garden experimental design, 140 fish were PIT-tagged, vaccinated and distributed equally into 4 tanks (35 fish for each group, including a control group). Blood samples were taken every 2 weeks to evaluate IgM titers. At 10 weeks post-immunization, all groups were ip challenged with 100 times the calculated LD50 for A. salmonicida J410. A. salmonicida was detected after 5 days post-infection (dpi) in all collected tissues. At 30 days post-challenge the relative percentage survival (RPS) with respect to the control group was calculated for each vaccine. The RPS for the bacterin mix was 65.22%, for Forte Micro 4® vaccine was 56.52% and for Alpha Ject Micro 4® was 30.43%, and these RPS values were reflected by A. salmonicida tissue colonization levels at 10 days post-challenge. Total IgM titers peaked at 6-8 weeks post-immunization, where the autogenous vaccine group showed the highest IgM titers and these values were consistent with the RPS data. Also, we determined that the A. salmonicida A-layer binds to immunoglobulins F(ab)' in a non-specific fashion, interfering with immune assays and potentially vaccine efficacy. Our results indicate that vaccine design influences sablefish immunity and provide a guide for future sablefish vaccine programs.

Vibrogen-2 vaccine trial in lumpfish (Cyclopterus lumpus) against Vibrio anguillarum.

Lumpfish (Cyclopterus lumpus), a native fish of the North Atlantic Ocean, is utilized as cleaner fish to biocontrol sea lice infestations in Atlantic salmon aquaculture. However, bacterial infections are affecting cleaner fish performance. Vibrio anguillarum, the aetiological agent of vibriosis, is one of the most frequent bacterial infections in lumpfish, and effective vaccine programmes against this pathogen have been identified as a high priority for lumpfish. Vibrogen-2 is a commercial polyvalent bath vaccine that contains formalin-inactivated cultures of V. anguillarum serotypes O1 and O2, and Vibrio ordalii. In this study, we evaluated Vibrogen-2 efficacy in lumpfish against a local isolated V. anguillarum strain. Two groups of 125 lumpfish were bath-immunized, bath-boost-immunized at four weeks post-primary immunization, and intraperitoneally (i.p.) boost-immunized at eight weeks post-primary immunization. The control groups were i.p. mock-immunized with PBS. Twenty-seven weeks post-primary immunization, the fish were i.p. challenged with 10 or 100 times the V. anguillarum J360 LD50 dose. After the challenge, survival was monitored daily, and samples of tissues were collected at ten days post-challenge. Commercial vaccine Vibrogen-2 reduced V. anguillarum tissue colonization and delayed mortality but did not confer immune protection to C. lumpus against the V. anguillarum i.p. challenge.

MiRNA-513a-5p inhibits progesterone receptor expression and constitutes a risk factor for breast cancer: the hOrmone and Diet in the ETiology of breast cancer prospective study.

MicroRNAs (miRNAs) might be considered both predictors and players of cancer development. The aim of the present report was to investigate whether many years before the diagnosis of breast cancer miRNA expression is already disregulated. In order to test this hypothesis, we compared miRNAs extracted from leukocytes in healthy women who later developed breast cancer and in women who remain healthy during the whole 15-year follow-up time. Accordantly, we used a case-control study design nested in the hOrmone and Diet in the ETiology of breast cancer (ORDET) prospective cohort study addressing the possibility that miRNAs can serve as both early biomarkers and components of the hormonal etiological pathways leading to breast cancer development in premenopausal women. We compared leukocyte miRNA profiles of 191 incident premenopausal breast cancer cases and profiles of 191 women who remained healthy over a follow-up period of 20 years. The analysis identified 20 differentially expressed miRNAs in women candidate to develop breast cancer versus control women. The upregulated miRNAs, miR-513-a-5p, miR-513b-5p and miR-513c-5p were among the most significantly deregulated miRNAs. In multivariate analysis, miR-513a-5p upregulation was directly and statistically significant associated with breast cancer risk (OR = 1.69; 95% CI 1.08-2.64; P = 0.0293). In addition, the upregulation of miR-513-a-5p displayed the strongest direct association with serum progesterone and testosterone levels. The experimental data corroborated the inhibitory function of miR-513a-5p on progesterone receptor expression confirming that progesterone receptor is a target of miR-513a-5p. The identification of upregulated miR-513a-5p with its oncogenic potential further validates the use of miRNAs as long-term biomarker of breast cancer risk.

Expression of inhibitory regulators of innate immunity in patients with active tuberculosis.

BACKGROUND: Tuberculosis (TB) is an important cause of morbidity and mortality worldwide. Toll-like-receptors (TLRs) are important for the recognition of the causative agent Mycobacterium tuberculosis. Negative regulation of TLRs is necessary to control deleterious inflammatory damage, but could provide a means of immune evasion by M. tuberculosis as well. METHODS: To obtain insight in the extent of expression of inhibitory regulators of immunity in patients with active TB, peripheral-blood-mononuclear-cells (PBMCs) and plasma were obtained from 54 TB patients and 29 healthy blood donors from Chittagong, Bangladesh. Bilateral alveolar macrophages were obtained from an infected versus a contralateral normal lung segment of 9 patients. Statistical analyses were performed using Mann-Whitney U and Wilcoxon matched pairs testing. Correlations were calculated using the Spearman rho test. RESULTS: PBMCs harvested from TB patients demonstrated increased mRNA expression of IL-1-receptor-associated-kinase-M, suppressor-of-cytokine-signalling-3 and Toll-interacting-protein. Flow cytometry revealed enhanced expression of IL-1-receptor-like-1 (ST2) on lymphocytes. Plasma soluble ST2 was elevated in patients with TB and correlated with established TB biomarkers, most strongly with soluble interleukin-2 receptor subunit α and interleukin-8. Alveolar macrophage mRNA expression of negative TLR regulators did not differ between the infected and contralateral lung side. CONCLUSION: These results show enhanced expression of distinct negative regulators of innate immunity in PBMCs of patients with TB and identify plasma soluble ST2 as a potential novel biomarker for TB disease activity.