Production of monoclonal antibody based on HA and NP protein of bat H18N11 influenza virus in mice

Master of ScienceDepartment of Diagnostic Medicine/PathobiologyWenjun MaInfluenza A viruses (IAVs) usually circulate in the waterfowl, but some of them with the capability to cross the species-specific boundaries and infect the mammalian host can cause severe zoonotic outbreaks. The sequences of H17N10 and H18N11 subtypes of bat IAVs have been identified, but the resource of these bat IAVs are still limited. Among eight proteins of bat IAVs, the surface glycoprotein HA and internal protein NP are important and can act as immunogenic proteins. They can be a good target for further research to study bat IAVs. Therefore, in this study, we produced a panel of NP and HA protein-specific monoclonal antibodies (mAbs) of bat H18N11 influenza virus. We immunized the BALB/C mice with baculovirus expressed HA and NP protein and then produced hybridoma cells by fusing spleen B cells with myeloma SP2/0 cells. We further tested these mAbs to identify the characteristics by using different immune assays. Six NP protein-derived mAbs were found specific and weakly bound with NP protein of H18N11 in immunofluorescence assay (IFA). These mAbs only reacted with NP protein of H18N11 virus other than those of conventional IAVs in IFA. In contrast, all the HA-specific mAbs failed to be reactive with HA of H18N11 in IFA. Isotyping of the mAbs was characterized by ELISA, and the results showed that all the NP-specific mAbs belong to IgG3. These NP-protein-derived mAbs specifically recognized the possible epitope with the amino acid sequence of position 445-458. To our surprise, all the NP-specific mAbs were not found reactive in the western blotting (WB) assay to detect the NP protein of H18N11 virus. Though the NP-specific mAbs were not specific in WB, they could detect NP of H18N11 by using IFA and ELISA. Therefore, the panel of NP mAbs can be a useful tool to study bat IAVs

Periodicity in the levels of serum plasminogen activator inhibitor-1 is a robust prognostic factor for embryo implantation and clinical pregnancy in ongoing IVF cycles

Context: Plasminogen activator inhibitor-1 (PAI-1) has been inversely correlated to proteolytic extracellular-matrix degradation exerted by urokinase-type (u-PA) and tissue-type plasminogen activators (t-PA). Any pathological disturbance in PAI-1 levels may lead to several pregnancy complications. Aims: To assess the influence of periodicity in serum PAI-1 levels on embryo implantation and clinical pregnancy outcome in IVF cycles Settings and Design: Prospective study of 120 IVF cycles at private infertility centre. Material and Methods: Endometrial response (ER) assessment by measuring Endometrial thickness (cm) and echopattern (grade). Serum PAI-1(ng/ml) measurement by ELISA method on day of hCG, day of ET and days 7 and 14 of ET. Main outcome measure was clinical pregnancy. Statistical Analysis: Student "t" test, ANOVA, Post-test for linear trend, Pearson Correlation. Results: PAI-1 levels declined from dhCG to dET (318.8 ± 36.1 to 176.1 ± 28.4) whereas they increased steadily from dET to d7 to d14ET (176.1 ± 28.4 to 285.2 ± 30.4 to 353.5 ± 150.4;P = 0.0004) in pregnant group (n = 31). Conversely, dhCG to dET levels increased in both nonpregnant (n = 75; 173.8 ± 18.3 to 280.8 ± 26.1) and biochemical pregnancy BCP (n = 14; 172.7 ± 31.1 to 216 ± 30.1) groups. The rising pattern from dET to d7 to d14ET was not observed in non-pregnant and BCP groups. ER thickness and grade shared significant correlation with serum PAI-1 on dET (Pearson r: ER = 0.28, Grade = 0.29) and d7ET (Pearson r: ER = 0.40, Grade = 0.23). Conclusions: Periodicity in serum PAI-1 levels offers a robust prognostic factor for predicting clinical pregnancy outcome. The dhCG to dET PAI-1 transition is a decisive factor for either transferring embryos in same/ongoing cycle or cryopreserving them and postponing ET to subsequent natural cycle

Follicular fluid dehydroepiandrosterone sulfate is a credible marker of oocyte maturity and pregnancy outcome in conventional in vitro fertilization cycles

Aim: To investigate if the level of dehydroepiandrosterone sulfate (DHEA-s) in follicular fluid (FF) influences the competence of oocytes to fertilize, develop to the blastocyst stage, and produce a viable pregnancy in conventional in vitro fertilization (IVF) cycles. Settings and Design: Prospective study of age-matched, nonpolycystic ovary syndrome (PCOS) women undergoing antagonist stimulation protocol involving conventional insemination and day 5 blastocyst transfer. Materials and Methods: FF levels of DHEA-s and E2 were measured by a radio-immuno-assay method using diagnostic kits. Fertilization rate, embryo development to the blastocyst stage and live birth rate were main outcome measures. Cycles were divided into pregnant/nonpregnant groups and also into low/medium/high FF DHEA-s groups. Statistical analysis was done by GraphPad Prism V software. Results: FF DHEA-s levels were significantly higher in pregnant (n = 111) compared to nonpregnant (n = 381) group (1599 ± 77.45 vs. 1372 ± 40.47 ng/ml; P = 0.01). High (n = 134) FF DHEA-s group had significantly higher percentage of metaphase II (MII) oocytes (91.5 vs. 85.54 vs. 79.44%, P < 0.0001), fertilization rate (78.86 vs. 74.16 vs. 71.26%, P < 0.0001), cleavage rate (83.67 vs. 69.1 vs. 66.17%, P = 0.0002), blastocyst formation rate (37.15 vs. 33.01 vs. 26.95%, P < 0.0001), and live birth rate (29.85 vs. 22.22 vs. 14.78%, P = 0.017) compared to medium (n = 243) and low (n = 115) FF DHEA-s groups, respectively despite comparable number of oocytes retrieved and number of blastocysts transferred. FF DHEA-s levels correlated significantly with the attainment of MII oocytes (Pearson r = 0.41) and fertilization rates (Pearson r = 0.35). Conclusion: FF DHEA-s level influences the oocyte maturation process and is predictive of fertilization, embryo development to the blastocyst stage and live birth rates in non-PCOS women undergoing conventional IVF cycles

Geographic disparities and temporal changes of COVID-19 incidence risks in North Dakota, United States

Abstract Background COVID-19 is an important public health concern due to its high morbidity, mortality and socioeconomic impact. Its burden varies by geographic location affecting some communities more than others. Identifying these disparities is important for guiding health planning and service provision. Therefore, this study investigated geographical disparities and temporal changes of the percentage of positive COVID-19 tests and COVID-19 incidence risk in North Dakota. Methods COVID-19 retrospective data on total number of tests and confirmed cases reported in North Dakota from March 2020 to September 2021 were obtained from the North Dakota COVID-19 Dashboard and Department of Health, respectively. Monthly incidence risks of the disease were calculated and reported as number of cases per 100,000 persons. To adjust for geographic autocorrelation and the small number problem, Spatial Empirical Bayesian (SEB) smoothing was performed using queen spatial weights. Identification of high-risk geographic clusters of percentages of positive tests and COVID-19 incidence risks were accomplished using Tango’s flexible spatial scan statistic. ArcGIS was used to display and visiualize the geographic distribution of percentages of positive tests, COVID-19 incidence risks, and high-risk clusters. Results County-level percentages of positive tests and SEB incidence risks varied by geographic location ranging from 0.11% to 13.67% and 122 to 16,443 cases per 100,000 persons, respectively. Clusters of high percentages of positive tests were consistently detected in the western part of the state. High incidence risks were identified in the central and south-western parts of the state, where significant high-risk spatial clusters were reported. Additionally, two peaks (August 2020-December 2020 and August 2021-September 2021) and two non-peak periods of COVID-19 incidence risk (March 2020-July 2020 and January 2021-July 2021) were observed. Conclusion Geographic disparities in COVID incidence risks exist in North Dakota with high-risk clusters being identified in the rural central and southwest parts of the state. These findings are useful for guiding intervention strategies by identifying high risk communities so that resources for disease control can be better allocated to communities in need based on empirical evidence. Future studies will investigate predictors of the identified disparities so as to guide planning, disease control and health policy

An overview of caspase: Apoptotic protein for silicosis

Silicosis is a chronic lung disease characterized by granulomatous and fibrotic lesions, which occurs due to accumulation of respirable silica mineral particles. Apoptosis is an important phenomenon of cell death in silicosis. The relationship between silica dust and its exposure is well established. But, the complex chain of cellular responses, which leads to caspase activation in silicosis, has not been fully discovered. Caspase activation plays a central role in the execution of apoptosis. Silica-induced apoptosis of the alveolar macrophages could potentially favor a proinflammatory state, occurring in the lungs of silicotic patients, resulting in the activation of caspase prior to induction of the intrinsic and extrinsic apoptosis pathways. Recent studies indicated that apoptosis may involve in pulmonary disorders. This review summarizes the current knowledge about the underling mechanism of biochemical pathways in caspase activation that have been ignored so far in silicosis. In addition, caspase could be a key apoptotic protein that can be used as an effective biomarker for the study of occupational diseases. It may provide an important link in understanding the molecular mechanisms of silica-induced lung pathogenesis

Follicular fluid insulin like growth factor-1 (FF IGF-1) is a biochemical marker of embryo quality and implantation rates in in vitro fertilization cycles

Context: Insulin-like growth factor-1 (IGF-1) has been reported to play a role in human follicular and embryonic development. However, earlier studies carried out mostly in animal models or in culture mediums supplemented with IGF-1 have been unable to directly link IGF-1 with embryo quality. Results correlating IGF-1 with pregnancy outcome have also been ambiguous so far. Aim: The aim of this study is to find if in situ follicular-fluid level of IGF-1 is predictive of embryo quality and implantation rates in in vitro fertilization (IVF) cycles. Settings and Design: Prospective study involving 120 cycles of conventional IVF-embryo transfer in infertile women. Subjects and Methods: IGF-1 concentrations were estimated in pooled follicular-fluid on the day of oocyte-pickup. Embryo quality was assessed daily at different developmental stages. Cycles were sorted into low and high follicular fluid insulin-like growth factor-1 (FF IGF-1) groups according to the median value of measurement. Embryo quality, clinical pregnancy and implantation rate were the main outcome measures. Statistical Analysis: Graph-pad Prism 5 statistical package. Results: FF IGF-1 correlates with embryo quality (Pearson r = 0.3894, r2 = 0.1516, P 58.50 ng/mg protein (receiver operating characteristics AUC : 0.85 ± 0.03, 95% CI: 0.78-0.91). Conclusion: FF IGF-1 is a plausible biochemical marker of embryo quality and implantation rate and correlates with clinical pregnancy rates in conventional IVF cycles

Prevalence of Eimeria spp. with associated risk factors in dairy calves in Sylhet, Bangladesh

Abstract Background Bovine eimeriosis is thought to be very important for the productivity and health of cattle all over the world. Despite the importance of cattle farming in Sylhet, little is known about the prevalence of bovine Eimeria spp. and the risk factors connected with it. Objectives We conducted a study to evaluate the prevalence, species diversity and associated risk factors of Eimeria spp. in a population of 50 cattle farms from 12 upazilas (sub‐district) in Sylhet district. Methods Faecal samples were collected randomly from a total of 554 calves ranging in age from 1 month to 2 years old during a period of 7 months. We used Flotation and McMaster techniques for parasitological examination. Species identification was done by using their morphological and morphometric characteristics. Results Out of 554 calves, 308 were found to be positive for Eimeria species (55.60%). Seven species of Eimeria were identified. Among the identified species, E. bovis (38.98%), E. zuernii (26.17%) and E. alabamensis (22.38%) were found to be the most prevalent species. Mixed and species‐specific Eimeria infection were (24.73%; 95% CI 21.32–28.49) and (30.87%; 95% CI 27.17–34.84), respectively. In addition, the highest prevalence was observed at Zakigonj (68%; 95% CI 58.34–76.33) and the lowest at Companygonj (40%; 95% CI 30.94–49.80). Eimeria species intensity ranged between 50 and 76,550 oocyst per gram of faeces. Analysis of associated risk factors by using multivariate logistic regression analysis revealed that age, gender and body condition were significantly (p < 0.05) associated with Eimeria infection. Conclusions Based on these present findings, it can be assumed that ‘coccidia belong to the most prevalent pathogens in the population of calves in the study area’. Thus, the findings of this study could be used as tools for adoptive surveillance and effective control and prevention of the disease in cattle populations in this region

Additional file 1 of Geographic disparities and temporal changes of COVID-19 incidence risks in North Dakota, United States

Additional file 1. The data used in this study have been provided as supporting information files together with the submission