Health care providers' preparedness and health care protection against the third wave of COVID-19 pandemics in a resource-limited setting in Southwest Ethiopia: a multi-center cross-sectional study.

Introduction: the provision of quality health care during the COVID-19 pandemic depends largely on the health of health care providers. Health care providers as frontline caregivers dealing with infected patients play a significant role in limiting the outbreak of the disease by implementing safety and prevention practices. However, low and middle-income countries experience barriers to preparedness due to limited resources. Methods: an institutional-based cross-sectional study was conducted among 326 health care providers from August 10-25, 2021 in Gurage zonal public hospitals. A simple random sampling technique was used to select the study participants. A pretested self -administered structured questionnaire was used as a data collection tool. The data were entered into the Epi-data 3.1 and exported to Statistical package for the social sciences (SPSS) version 25.0 for analysis. Both descriptive statistics and inferential statistics were presented. Results: this study showed that 53.1%, of health care providers, had adequate preparation against COVID-19 pandemics. The finding showed that monthly income, occupation, and working experience were found to be significantly associated with health care providers' preparedness. Nearly one-quarter (24.8%), 28.3%, 34.5%, and 39.8% of health care providers had access to facemasks, alcohol sanitizer, glove, and isolation gowns respectively. Conclusion: the levels of health care providers' preparedness and health care protection against the third wave COVID-19 pandemic were found to be low. Based on our findings, the government and other stakeholders should design interventions to increase health care providers' preparedness to respond to the ongoing pandemic and purchase an adequate supply of personal protective equipment to protect the health care providers.

Anemia and its predictors among chronic kidney disease patients in Sub-Saharan African countries: A systematic review and meta-analysis.

INTRODUCTION: Anemia is a serious complication of chronic kidney disease (CKD) with a significant adverse outcome on the burden and progression of the disease. Hence, the study intended to assess the pooled prevalence of anemia and its predictors among CKD patients in Sub-Saharan African nations. METHODS: To identify the relevant studies systematic searches were carried out in Medline, EMBASE, HINARI, Google Scholar, Science Direct, and Cochrane Library. From selected studies, data were taken out with a standardized data extraction format prepared in Microsoft Excel. Inverse variance (I2) tests were employed to evaluate the heterogeneity across the included studies. Due to substantial heterogeneity among the studies, a random-effects meta-analysis technique was employed to estimate the pooled prevalence of anemia. Subgroup analysis, sensitivity analysis, and meta-regression analysis were carried out to search the possible bases of heterogeneity. Funnel plot symmetry, Begg's test, and Egger's regression test were employed to assess the existence of publication bias. In addition, factors associated with anemia among CKD patients were examined. All statistical analyses were carried out with STATA™ Version 14 software. RESULTS: A total of 25 studies with 5042 study participants were considered in this study. The pooled prevalence of anemia among CKD patients was estimated to be 59.15% (95% CI, 50.02-68.27) with a substantial level of heterogeneity as evidenced by I2 statistics (I2 = 98.1%; p < 0.001). Stage of CKD (3-5) (pooled odds ratio (POR) = 5.33, 95% CI:4.20-6.76), presence of diabetes mellitus (POR = 1.75, 95% CI: 1.10-2.78), hemodialysis history (POR = 3.06, 95% CI: 1.63-5.73), and female sex (POR = 2.50, 95% CI: 1.76-3.55) were significantly related with anemia. CONCLUSIONS: More than half of CKD patients were suffering from anemia. Stage of CKD, presence of DM, hemodialysis history, and being female sex were factors associated with anemia among CKD patients.

Attitudes towards receiving COVID-19 vaccine and its associated factors among Southwest Ethiopian adults, 2021.

INTRODUCTION: Many countries around the world are still affected by the global pandemic of coronavirus disease. The vaccine is the most effective method of controlling Coronavirus Disease 2019 (COVID-19). However, attitudes toward vaccination are heavily affected by different factors besides vaccine availability. OBJECTIVES: This study aimed to determine community attitudes toward the COVID-19 vaccine in Gurage Zone, Ethiopia. METHODS: A community-based cross-sectional study was conducted from November 15th to December 15th, 2021. A simple random sampling technique was used to select 364 participants in the study area. An interview-administered structured questionnaire was used to collect the data; the data was entered into Epidata 3.1 version, and then exported to SPSS version 23 for further analysis. Descriptive statistics were used to determine the characteristics of study participants. Binary and multivariable logistic regression analyses with a p-value of less than 0.05 were used as a measure of significance. RESULTS: In this study, 44.7% of study participants had a favorable attitude toward the COVID-19 vaccine. Perceived potential vaccine harm [AOR: 1.85; 95% CI (1.15-2.96)], Having ever had a chronic disease [AOR: 3.22; 95% CI (2.02-5.14)], community belief on the effectiveness of the vaccine [AOR: 2.02; 95% CI (1.27-3.22)], and average monthly income 3001-5000 ETB [AOR: 0.54; 95% CI (0.30-0.97)], average monthly income 5001-10000 ETB [AOR: 0.48; 95% CI(0.27-0.86)] were statistically significantly towards COVID-19 vaccination. CONCLUSIONS: Overall, less than half of the participants had a favorable attitude toward the COVID-19 vaccine. Perceived potential vaccine harm, having ever had a chronic disease, community belief in the effectiveness of the vaccine, and average monthly income were determinant factors of the community's attitude toward COVID-19 vaccination. As a result, information conversation with the community's awareness of the COVID-19 vaccination in reducing vaccine-related suspicion.

Self-care practice and its associated factors among diabetic patients attending public hospitals in Gurage zone southwest, Ethiopia.

BACKGROUND: Diabetes is a chronic disease that requires lifelong medical treatment and lifestyle modifications. Even though patients often neglect their own needs, self-care is an important factor in preventing and delaying complications related to diabetes. There are limited studies about self-care practice, and most of the studies conducted in Ethiopia focused on some parts of the recommended self-care practice. Therefore, this study aimed to assess the self-care practice and associated factors among diabetic patients in Gurage zone, south Ethiopia. METHODS: An institution-based cross-sectional study was conducted from February 6 to March 29, 2021. A systematic sampling method was employed to select 420 study participants. The data were collected using a pretested interviewer-administered questionnaire. All variables with P < 0.25 in the bi-variable logistic regression analysis were entered into multivariable logistic regression analysis. The statistical significance was declared at a p-value < 0.05. RESULTS: A total of 384 diabetes patients participated with a response rate of 91.4%. This study showed that more than half (60.4%) of the study participants had poor self-care practices. Being female (AOR: 2.40; 95% CI:1.31-4.40), rural residence (AOR:7.16;95% CI: 3.31-15.46), duration of diabetes treatment 5-10 years (AOR: 0.03; 95% CI: 0.1-0.11), duration of diabetes treatment ≥ 10 years (AOR:0.8; 95% CI: 0.03-0.21), haven't social support (AOR: 0.10; 95% CI: 0.05-0.23), haven't got health education (AOR: 0.17,95%CI 0.09-0.32) were factors significantly associated with self-care practice. CONCLUSIONS: Despite, the importance of diabetes self-care practice for the management of diabetes and preventing its complications, a high number of diabetes patients had poor self-care practices. Female, rural residence, duration of diabetes mellitus, lack of social support, and not get of health education were significantly associated with poor self-care practice. Therefore, health care providers should give attention to diabetic patients with the aforementioned factors that affect diabetic patients' self-care practices.

Lifespan and Associated Factors of Peripheral Intravenous Cannula Among Hospitalized Children in Public Hospitals of the Gurage Zone, Ethiopia, 2021.

Background: Peripheral intravenous cannulas are routinely used in hospital-admitted children requiring intravenous therapy. The majority of peripheral IVC lines are removed before completion of therapy due to cannula complications in children. Peripheral intravenous cannula securing is a painful procedure for children, so recognizing the variables associated with the peripheral intravenous cannula lifespan would help decrease the complications and increase the duration of cannula patency. Objective: The main aim of this study was to determine the lifespan and associated factors of peripheral intravenous cannula among hospitalized children in Gurage zone public hospitals, Ethiopia, 2021. Methods: We conducted an institution-based cross-sectional study design among 422 admitted children in public hospitals of the Gurage zone using a systematic random sampling technique. We collected data from interviews of parents using structured questionnaires and direct observations using checklists. The data was coded and entered into EPI-DATA version 3.1 and exported to SPSS version 25 for analysis. Bivariable and multivariable analysis was used by using a binary logistic regression model. Finally, the variables with a p-value of <0.05 with a 95% confidence interval (CI) from the multivariable analysis were considered statistically significant. Results: One hundred and sixty-six children (41.4%) had a short cannula lifespan (below 30 h). Multivariable logistic regression analysis showed that neonatal intensive care unit (NICU) [AOR = 4.975; 95% CI (2.811-8.805)], reason for removal (complication) [AOR = 3.277; 95% CI (1.924-5.583)], fluid [AOR = 2.285; 95% CI (1.274-4.100)], and blood transfusion [AOR = 2.407; 95% CI (1.005-5.572)] were the statistically significant variables associated with the lifespan of a peripheral intravenous cannula. Conclusion and Recommendation: The proportion of the short lifespan of the peripheral intravenous cannula was higher in hospitalized children and health care providers better to use low concentration electrolytes, low osmotic pressure, and weak alkalinity fluid. The cannula access might be the responsibility of the health-trained staff, and conserving immediate removal upon the presence of a signal that indicates a complication.

Levels and predictors of anxiety, depression, and stress during COVID-19 pandemic among frontline healthcare providers in Gurage zonal public hospitals, Southwest Ethiopia, 2020: A multicenter cross-sectional study.

INTRODUCTION: The provision of quality health care during the COVID-19 pandemic depends largely on the health of health care providers. However, healthcare providers as the frontline caregivers dealing with infected patients, are more vulnerable to mental health problems. Despite this fact, there is scarce information regarding the mental health impact of COVID-19 among frontline health care providers in South-West Ethiopia. OBJECTIVE: This study aimed to determine the levels and predictors of anxiety, depression, and stress during the COVID-19 pandemic among frontline healthcare providers in Gurage zonal public hospitals, Southwest Ethiopia, 2020. METHODS: An institutional-based cross-sectional study was conducted among 322 health care providers from November 10-25, 2020 in Gurage zonal health institutions. A simple random sampling technique was used to select the study participants. A pretested self -administered structured questionnaire was used as a data collection technique. The data were entered into the Epi-data version 3.01 and exported to SPSS version 25.0 for analysis. Both descriptive statistics and inferential statistics (chi-square tests) were presented Bivariable and Multivariable logistic regression analyses were made to identify variables having a significant association with the dependent variables. RESULTS: The results of this study had shown that the overall prevalence of anxiety, depression and stress among health care providers during the COVID-19 pandemic was 36%, [95% CI = (30.7%- 41.3%)], 25.8% [95% CI = (21.1%- 30.4%)] and 31.4% [95% CI = (26.4%- 36.0%)] respectively. Age, Adjusted Odds Ratio [AOR = 7.9], Educational status, [AOR = 3.2], low monthly income [AOR = 1.87], and presence of infected family members [AOR = 3.3] were statistically associated with anxiety. Besides this, gender, [AOR = 1.9], masters [AOR = 10.8], and degree holder [AOR = 2.2], living with spouse [AOR = 5.8], and family [AOR = 3.9], being pharmacists [AOR = 4.5], and physician [AOR = (0.19)], were found to be statistically significant predictors of depression among health care providers. Our study finding also showed that working at general [AOR = 4.8], and referral hospitals [AOR = 3.2], and low monthly income [AOR = 2.3] were found to be statistically significant predictors of stress among health care providers. CONCLUSION: Based on our finding significant numbers of healthcare providers were suffered from anxiety, depression, and stress during the COVID-19 outbreak. So, the Government and other stakeholders should be involved and closely work and monitor the mental wellbeing of health care providers.

Association of glycemic control and anthropometric measurement among type 2 diabetes mellitus: a cross-sectional study.

INTRODUCTION: Maintaining good glycemic control is the main therapeutic target for diabetes mellitus (DM) patients. Nevertheless, due to complex factors, the majority of patients worldwide persist poorly controlled. Hence the study was conducted to determine the association of anthropometric measurement and other factors with glycemic control among type 2 DM patients. METHODS: An institutional-based cross-sectional study was conducted among 366 type 2 DM patients from October-1 to November 30/2019. Data were collected by a pre-tested structured questionnaire and data abstraction format. The collected data were edited in Epi-data-4.4.1.0 and exported to SPSS-25 for analysis. The degree of association was assessed using logistic regression analysis and expressed in terms of p value and odds ratio with a 95%CI. p value < 0.05 was considered statistically significant. RESULTS: Of the total 366 participants, (66.1%) had poor glycemic control. Increased waist circumference (AOR 2.37, 95% CI 1.28-4.40 for overweight and AOR 3.31, 95% CI 1.28-8.58 for obesity), long duration of DM (AOR 2.12, 95% CI 1.14-3.95 for 5-10 years and AOR 5.67, 95% CI 1.76-11.30 for > 10 years), occupation (AOR 3.74, 95% CI 1.94-7.23 for government employee and AOR 4.90, 95% CI 2.19-10.94 for private employee), and family history of DM (AOR 1.93, 95% CI 1.08-3.43), were significantly associated with poor glycemic control. CONCLUSION: Nearly two-thirds of study participants had poor glycemic control. Increased waist circumference, long duration of DM, occupational status, and family history of DM were independently associated with poor glycemic control. Therefore, type 2 DM patients with these identified factors should be strictly monitored to maintain good glycemic control.

Knowledge, Attitude, and Practices of Parents About Immunization of Infants and Its Associated Factors in Wadla Woreda, North East Ethiopia, 2019.

INTRODUCTION: Improving infant immunization coverage and timeliness is a key health policy objective in many developing countries such as Ethiopia. Despite this, full immunization coverage in Ethiopia becomes low with the concurrent significant burden of vaccine-preventable diseases among infants. A previously published study acknowledged that parental knowledge, attitude, and practice towards infant immunization are vital issues to improve coverage and influence uptake. OBJECTIVE: To assess parents' knowledge, attitude, practice, and its associated factors regarding immunization of infants at Wadla Woreda, North East Ethiopia, 2019. MATERIAL AND METHODS: A community-based cross-sectional study was employed among 418 parents from March to April 2019 in Wadla Woreda, North East Ethiopia. A stratified sampling technique was used to approach the study subjects. An interviewer-administered structured questionnaire was used. The collected data were cleaned, coded, and entered in EPI-Info 7.2 and transferred to SPSS version 25.0 for analysis. Binary logistic regression analyses were performed to identify variables having a significant association with parental knowledge, attitude, and practice. Variables with a p-value of ≤0.05 and AOR with 95% CI were declared as having a statistically significant association during multivariable logistic regression analysis. RESULTS: In this study, 65.1%, 57.3%, and 55.3% of the parents had good knowledge, a favorable attitude, and good practice towards infant immunization, respectively. Parent's educational status (AOR=5.330), urban residency (AOR=2.788), favorable attitude (AOR=4.308) and got immunization service two-three times (AOR=3.227) and four-five times (AOR=2.254) were statistically associated with knowledge of parents. Parents who attend primary school [AOR=0.451) and secondary school [AOR=0.320), parents who were mothers of the child [AOR=3.813), and respondents who had good knowledge about infant immunization (AOR= 4.592) were significantly associated with a favorable parental attitude. Infant immunization practice was significantly associated with parental education who attend primary school (AOR=2.513), secondary school (AOR=2.546) and higher education (AOR=11.988), parents who had good knowledge of infant immunization (AOR= 4.206), and short waiting time (AOR=3.881). CONCLUSION AND RECOMMENDATION: Parental KAP towards infant immunization was found to be lower than most of the study findings quoted in this study. Improving the knowledge, attitude, and practice of parents about immunization and vaccine-preventable diseases was recommended by providing health education and health promotion interventions.

Platelet Indices and Its Association with Microvascular Complications Among Type 2 Diabetes Mellitus Patients in Northeast Ethiopia: A Cross-Sectional Study.

PURPOSE: Platelet indices have been shown to be associated with complications of diabetes mellitus (DM). However, the clinical relevance of platelet indices for assessing the risk of being microvascular complications among patients with DM is largely overlooked. Hence this study aimed to determine the association of platelet indices with microvascular complications among type 2 DM patients. PATIENTS AND METHODS: A hospital-based cross-sectional study was conducted among 352 type 2 DM patients from April 1 to May 30/2019 at Debre Berhan Referral Hospital. Data were collected by interview, physical measurements, document review, and laboratory tests. Then the data were entered into Epi-data manager-4.4.1.0 and exported into SPSS-25 for analysis. Platelet indices were compared between participants with and without microvascular complications using an independent t-test. A logistic regression model was used to assess the association between microvascular complications and platelet indices. All inferences were considered statistically significant at P<0.05. RESULTS: One hundred seventeen (33.2%) of participants were suffering from at least one microvascular complications. The mean platelet volume (MPV), platelet distribution width (PDW), and platelet-large cell ratio (P-LCR) were significantly increased in DM patients with complications as compared to without complications (13.57±2.17fl vs. 11.76±1.93fl), (16.57±2.49fl vs. 14.97±2.41fl) and (28.09±7.58% vs. 24.19±6.46%), respectively (all are P˂0.001). In multivariate logistic regression analysis, MPV (AOR=1.68, 95% CI: 1.37-2.05), PDW (AOR=1.37, 95% CI: 1.15-1.63), P-LCR (AOR=1.07, 95% CI: 1.01-1.14), age (AOR=1.07, 95% CI: 1.01-1.12), and duration of DM (AOR=1.31, 95% CI: 1.18-1.46) were significantly associated with the presence of microvascular complications. CONCLUSION: MPV, PDW, and P-LCR were significantly higher among type 2 DM patients with microvascular complications. This indicates that platelet indices can be used as prognostic markers of vascular complications and can be used as simple and cost-effective parameters to predict microvascular complications particularly in patients with old age and long duration of DM.

DNA methylation dynamics at imprinted genes during bovine pre-implantation embryo development.

BACKGROUND: In mammals, maternal differentially methylated regions (DMRs) acquire DNA methylation during the postnatal growth stage of oogenesis, with paternal DMRs acquiring DNA methylation in the perinatal prospermatagonia. Following fusion of the male and female gametes, it is widely accepted that murine DNA methylation marks at the DMRs of imprinted genes are stable through embryogenesis and early development, until they are reprogrammed in primordial germ cells. However, the DNA methylation dynamics at DMRs of bovine imprinted genes during early stages of development remains largely unknown. The objective of this investigation was to analyse the methylation dynamics at imprinted gene DMRs during bovine embryo development, from blastocyst stage until implantation. RESULTS: To this end, pyrosequencing technology was used to quantify DNA methylation at DMR-associated CpG dinucleotides of six imprinted bovine genes (SNRPN, MEST, IGF2R, PLAGL1, PEG10 and H19) using bisulfite-modified genomic DNA isolated from individual blastocysts (Day 7); ovoid embryos (Day 14); filamentous embryos (Day 17) and implanting conceptuses (Day 25). For all genes, the degree of DNA methylation was most variable in Day 7 blastocysts compared to later developmental stages (P < 0.05). Furthermore, mining of RNA-seq transcriptomic data and western blot analysis revealed a specific window of expression of DNA methylation machinery genes (including DNMT3A, DNMT3B, TRIM28/KAP1 and DNMT1) and proteins (DNMT3A, DNMT3A2 and DNMT3B) by bovine embryos coincident with imprint stabilization. CONCLUSION: The findings of this study suggest that the DNA methylation status of bovine DMRs might be variable during the early stages of embryonic development, possibly requiring an active period of imprint stabilization.

Amino acids in the uterine luminal fluid reflects the temporal changes in transporter expression in the endometrium and conceptus during early pregnancy in cattle.

In cattle, conceptus-maternal interactions are critical for the establishment and maintenance of pregnancy. A major component of this early interaction involves the transport of nutrients and secretion of key molecules by uterine epithelial cells to help support conceptus development during the peri-implantation period of pregnancy. Objectives were to: 1) analyze temporal changes in the amino acid (AA) content of uterine luminal fluid (ULF) during the bovine estrous cycle; 2) understand conceptus-induced alterations in AA content; 3) determine expression of AA transporters in the endometrium and conceptus; and 4) determine how these transporters are modulated by (Progesterone) P4. Concentrations of aspartic acid, arginine, glutamine, histidine, lysine, isoleucine, leucine, phenylalanine and tyrosine decreased on Day 16 of the estrous cycle but increased on Day 19 in pregnant heifers (P<0.05). Glutamic acid only increased in pregnant heifers on Day 19 (P<0.001). Asparagine concentrations were greater in ULF of cyclic compared to pregnant heifers on Day 7 (P<0.05) while valine concentrations were higher in pregnant heifers on Day 16 (P<0.05). Temporal changes in expression of the cationic AA transporters SLC7A1 SLC7A4 and SLC7A6 occurred in the endometrium during the estrous cycle/early pregnancy coordinate with changes in conceptus expression of SLC7A4, SLC7A2 and SLC7A1 (P<0.05). Only one acidic AA transporter (SLC1A5) increased in the endometrium while conceptus expression of SLC1A4 increased (P<0.05). The neutral AA transporters SLC38A2 and SLC7A5 increased in the endometrium in a temporal manner while conceptus expression of SLC38A7, SLC43A2, SLC38A11 and SLC7A8 also increased (P<0.05). P4 modified the expression of SLC1A1, -1A4, -1A5, -38A2, -38A4, -38A7, -43A2, -6A14, -7A1, -7A5 and -7A7 in the endometrium. Results demonstrate that temporal changes in AA in the ULF reflect changes in transporter expression in the endometrium and conceptus during early pregnancy in cattle, some of which are modified by P4.

Proteomic analysis of uterine fluid during the pre-implantation period of pregnancy in cattle.

The aims of this study were (i) to characterize the global changes in the composition of the uterine luminal fluid (ULF) from pregnant heifers during pregnancy recognition (day 16) using nano-LC MS/MS; (ii) to describe quantitative changes in selected proteins in the ULF from days 10, 13, 16 and 19 by Isobaric tags for Relative and Absolute Quantification (iTRAQ) analysis; and (iii) to determine whether these proteins are of endometrial or conceptus origin, by examining the expression profiles of the associated transcripts by RNA sequencing. On day 16, 1652 peptides were identified in the ULF by nano-LC MS/MS. Of the most abundant proteins present, iTRAQ analysis revealed that RPB4, TIMP2 and GC had the same expression pattern as IFNT, while the abundance of IDH1, CST6 and GDI2 decreased on either day 16 or 19. ALDOA, CO3, GSN, HSP90A1, SERPINA31 and VCN proteins decreased on day 13 compared with day 10 but subsequently increased on day 16 (P<0.05). Purine nucleoside phosphorylase (PNP) and HSPA8 decreased on day 13, increased on day 16 and decreased and increased on day 19 (P<0.05). The abundance of CATD, CO3, CST6, GDA, GELS, IDHC, PNPH and TIMP2 mRNAs was greater (P<0.001) in the endometrium than in the conceptus. By contrast, the abundance of ACTB, ALDOA, ALDR, CAP1, CATB, CATG, GD1B, HSP7C, HSP90A, RET4 and TERA was greater (P<0.05) in the conceptus than in the endometrium. In conclusion, significant changes in the protein content of the ULF occur during the pre-implantation period of pregnancy reflecting the morphological changes that occur in the conceptus.

Predictive value of bovine follicular components as markers of oocyte developmental potential.

The follicle is a unique micro-environment within which the oocyte can develop and mature to a fertilisable gamete. The aim of this study was to investigate the ability of a panel of follicular parameters, including intrafollicular steroid and metabolomic profiles and theca, granulosa and cumulus cell candidate gene mRNA abundance, to predict the potential of bovine oocytes to develop to the blastocyst stage in vitro. Individual follicles were dissected from abattoir ovaries, carefully ruptured under a stereomicroscope and the oocyte was recovered and individually processed through in vitro maturation, fertilisation and culture. The mean (±s.e.m.) follicular concentrations of testosterone (62.8±4.8 ngmL(-1)), progesterone (616.8±31.9 ngmL(-1)) and oestradiol (14.4±2.4 ngmL(-1)) were not different (P>0.05) between oocytes that formed (competent) or failed to form (incompetent) blastocysts. Principal-component analysis of the quantified aqueous metabolites in follicular fluid showed differences between oocytes that formed blastocysts and oocytes that degenerated; l-alanine, glycine and l-glutamate were positively correlated and urea was negatively correlated with blastocyst formation. Follicular fluid associated with competent oocytes was significantly lower in palmitic acid (P=0.023) and total fatty acids (P=0.031) and significantly higher in linolenic acid (P=0.036) than follicular fluid from incompetent oocytes. Significantly higher (P<0.05) transcript abundance of LHCGR in granulosa cells, ESR1 and VCAN in thecal cells and TNFAIP6 in cumulus cells was associated with competent compared with incompetent oocytes.

Alterations in expression of endometrial genes coding for proteins secreted into the uterine lumen during conceptus elongation in cattle.

BACKGROUND: We hypothesized that genes that are up-regulated in the uterine endometrium at the initiation of conceptus elongation in cattle, and that encode for secreted proteins, contribute to the composition of the uterine luminal fluid (ULF) and ultimately, drive conceptus elongation. The aims of this study were to: 1) screen endometrial transcriptomic data for genes that encode secreted proteins on Day 13; 2) determine temporal changes in the expression of these genes during the estrous cycle/early pregnancy; 3) determine if expression of these genes is affected by altered concentrations of progesterone (P4) in vivo and 4) determine if the protein products of these genes are detectable in ULF. RESULTS: Of the fourteen candidate genes examined, quantitative real-time PCR analysis revealed the expression of APOA1, ARSA, DCN, LCAT, MUC13, NCDN, NMN, NPNT, NXPH3, PENK, PLIN2 and TINAGL1 was modulated in the endometrium (P<0.05) as the estrous cycle/early pregnancy progressed. APOA1, DCN and NPNT expression was higher in cyclic compared to pregnant heifers, and pregnancy increased (P<0.05) the expression of LCAT, NCDN, NMN, PLIN2 and TINAGL1. The magnitude of the increase in expression of APOA1, PENK and TINAGL1 on Day 13 was reduced (P<0.05) in heifers with low P4. Furthermore, low P4 decreased (P<0.05) the expression of LCAT and NPNT on Day 7, while an early increase (P<0.05) in the expression of NXPH3 and PLIN2 was observed in heifers with high P4. The protein products of 5 of the candidate genes (APOA1, ARSA, LCAT, NCDN and PLIN) were detected in the ULF on either Days 13, 16 or 19 of pregnancy. CONCLUSION: Using a candidate gene approach, we determined that both P4 concentration and the presence of the conceptus alter endometrial expression of PLIN2, TINAGL1, NPNT, LCAT, NMN and APOA1. Comparison of the expression profiles of these genes to proteins detected in ULF during conceptus elongation (i.e., Days 13 through 19) revealed the presence of APOA1, ARSA, LCAT, NCDN as well as members of the PLIN family of proteins that may play roles in driving conceptus elongation in cattle.

Towards rational use of antibiotics for suspected secondary infections in Buruli ulcer patients.

BACKGROUND: The emerging disease Buruli ulcer is treated with streptomycin and rifampicin and surgery if necessary. Frequently other antibiotics are used during treatment. METHODS/PRINCIPAL FINDINGS: Information on prescribing behavior of antibiotics for suspected secondary infections and for prophylactic use was collected retrospectively. Of 185 patients that started treatment for Buruli ulcer in different centers in Ghana and Bénin 51 were admitted. Forty of these 51 admitted patients (78%) received at least one course of antibiotics other than streptomycin and rifampicin during their hospital stay. The median number (IQR) of antibiotic courses for admitted patients was 2 (1, 5). Only twelve patients received antibiotics for a suspected secondary infection, all other courses were prescribed as prophylaxis of secondary infections extended till 10 days on average after excision, debridement or skin grafting. Antibiotic regimens varied considerably per indication. In another group of BU patients in two centers in Bénin, superficial wound cultures were performed. These cultures from superficial swabs represented bacteria to be expected from a chronic wound, but 13 of the 34 (38%) S. aureus were MRSA. CONCLUSIONS/SIGNIFICANCE: A guide for rational antibiotic treatment for suspected secondary infections or prophylaxis is needed. Adherence to the guideline proposed in this article may reduce and tailor antibiotic use other than streptomycin and rifampicin in Buruli ulcer patients. It may save costs, reduce toxicity and limit development of further antimicrobial resistance. This topic should be included in general protocols on the management of Buruli ulcer.

Transcriptomic changes in the bovine conceptus between the blastocyst stage and initiation of implantation.

Conceptus-maternal communication is vital for the successful establishment and maintenance of pregnancy, yet relatively little information exists for many of the mechanisms and the nature of the conceptus signals responsible for this cross-talk. Sub-optimal communication, resulting from impairment of conceptus development and/or from abnormal uterine receptivity, contributes to a high incidence of embryonic mortality. Therefore, detailed examination of the mechanisms regulating both pre- and peri-implantation conceptus development are necessary to fully understand the factors regulating successful post-hatching development, pregnancy recognition and implantation signaling. Despite significant progress in understanding of the temporal changes in the transcriptome of the uterine endometrium, there is only a rudimentary knowledge of the genes and pathways governing growth and development of the cattle conceptus. Furthermore, although there are a large number of studies describing gene expression profiles in bovine embryos focused mainly during the earlier preimplantation stages (up to and including Day 7), very little information exists for the post-hatching embryo and elongating conceptus. This period of development is arguably more important as a significant proportion of all embryonic loss occurs between Days 8 and 16 of pregnancy in cattle, corresponding to the time of hatching of the blastocyst from the zona pellucida and its subsequent elongation coincident with the time of maternal recognition of pregnancy. Given that this is a critical period in development leading up to maternal recognition and establishment of pregnancy, the identification of key genes and pathways regulating these crucial developmental events is essential.

Conceptus-endometrium crosstalk during maternal recognition of pregnancy in cattle.

Successful growth and development of the posthatching blastocyst and pregnancy establishment are a result of the interaction between a competent embryo and a receptive uterine environment. We examined the global transcriptome profiles of the Day 16 bovine conceptus and pregnant endometrium tissues using RNA-Seq to identify genes that contribute to the dialogue during the period of pregnancy recognition. Using stringent filtering criterion, a total of 16 018 and 16 262 transcripts of conceptus and pregnant endometrium origin, respectively, were identified with distinct tissue-specific expression profiles. Of these, 2261 and 2505 transcripts were conceptus and endometrium specific. Using Cytoscape software, a total of 133 conceptus ligands that interact with corresponding receptors on the endometrium and 121 endometrium ligands that interact with corresponding receptors on the conceptus were identified. While 87 ligands were commonly detected, 46 were conceptus specific and 34 endometrium specific. This study is one of the first to provide a comprehensive list of potentially secreted molecules in the conceptus that interact with receptors on the endometrium and vice versa during the critical window of maternal recognition of pregnancy. The identified tissue-specific genes may serve as candidates to study pregnancy recognition and they or downstream products may represent potential early markers of pregnancy.

Comparative analysis of nuclear transfer embryo-derived mouse embryonic stem cells. Part II: gene regulation.

In a mouse model nuclear transfer embryo-derived embryonic stem cell lines (ntESCs) of various genetic backgrounds and donor cell types were compared with reference ESCs and analyzed comprehensively at molecular level as a second part of a larger study. Expression profiles of ntESCs established by different NT-methods (piezoelectric microinjection or zona-free) were indistinguishable. However, expression profiling analyses identified differentially regulated genes between reference ESCs and ntESCs from different genetic backgrounds. A number of pluripotency and stemness marker genes significantly differed at the mRNA level between the cell lines. However, cluster and lineage analyses revealed that such differences had no effect on cell differentiation and cell fate. Regardless of the donor cell type, gene expression profiles of ntESCs were more similar to each other than to their counterpart fertilized embryo-derived ESCs of the same genotype. Overall, the results indicated that expression profile differences may be related to the genotype rather than to technical variations.

Comparative analysis of nuclear transfer embryo-derived mouse embryonic stem cells. Part I: cellular characterization.

Embryonic stem cells derived from nuclear transfer embryos (ntESCs) are particularly valuable for regenerative medicine, as they are a patient-specific and histocompatible cell source for the treatment of varying diseases. However, currently, little is known about their cellular and molecular profile. In the present study, in a mouse model different donor cell-derived ntESCs from various genetic backgrounds were compared with reference ESCs and analyzed comprehensively at the cellular level. A number of pluripotency marker genes were compared by flow cytometry and immunocytochemistry analysis. Significant differences at the protein level were observed for POU5F1, SOX2, FGF4, NANOG, and SSEA-1. However, such differences had no effect on in vitro cell differentiation and cell fate: derivatives of the three germ layers were detected in all ntESC lines. The neural and cardiac in vitro differentiation revealed minor differences between the cell lines, both at the mRNA and protein level. Karyotype analyses and cell growth studies did not reveal any significant variations. Despite some differences observed, the present study revealed that ntESC lines had similar differentiation competences compared to other ESCs. The results indicate that the observed differences may be related to the genotype rather than to the nuclear transfer technology.

RNA sequencing reveals novel gene clusters in bovine conceptuses associated with maternal recognition of pregnancy and implantation.

Successful establishment and maintenance of pregnancy can be attained only through optimum conceptus-maternal cross talk. Despite significant progress in our understanding of the temporal changes in the transcriptome of the uterine endometrium, we have only a rudimentary knowledge of the genes and pathways governing growth and development of the bovine conceptus. In particular, very little information exists for the posthatching embryo and elongating conceptus. This period of development is arguably the most important, as approximately 40% of all embryonic loss occurs between Days 8 and 17 of pregnancy in cattle. Here, we describe the global transcriptome profile of the bovine conceptus at five key stages of its pre- and peri-implantation growth (Days 7, 10, 13, 16, and 19) using state-of-the-art RNA sequencing techniques. More than 287 million reads were generated at the five stages, and more than 22 700 unique transcripts were detected. Analysis of variance followed by self-organizing maps identified differentially regulated (P < 0.05) genes organized in nine gene clusters forming a sequential transcript dynamics across these developmental stages. Of particular interest, genes in clusters 3 (n = 236) and 6 (n = 1409) were significantly up-regulated on Days 16 and 19, suggesting a role in maternal recognition and initiation of implantation. This transcriptome analysis of the bovine conceptus will provide a blueprint of the dynamic changes in gene expression occurring during maternal recognition and implantation and will complement existing knowledge of the temporal changes in the endometrial transcriptome, thus facilitating a better understanding of conceptus-maternal cross talk during the peri-implantation period of pregnancy.