Kit Manufacturers Research Articles

Angiotensin-II receptor type 1 and endothelin-1 receptor type A agonistic autoantibodies in patients with STEMI: new kids on the block

Abstract Background The Prognosis after ST-elevation myocardial infarction (STEMI) remains poor. Angiotensin-II and endothelin-1 contribute to adverse prognosis after STEMI through molecular mechanisms that lead to myocardial inflammation, fibrosis and, ultimately, adverse myocardial remodeling. Functional autoantibodies against angiotensin II type 1 (AT1R-AAs) and endothelin-1 type A (ETAR-AAs) receptors bind to the same receptors as natural ligands, eliciting similar (and amplified) responses. In patients with STEMI, AT1R-AAs and ETAR-AAs have been associated with a higher risk of developing microvascular obstruction and left ventricular remodeling. Both microvascular obstruction and left ventricular remodeling are associated with a poor prognosis. Purpose To assess the prognostic role of AT1R-AAs and ETAR-AAs after STEMI. Methods Consecutive STEMI patients who underwent primary percutaneous coronary intervention within 12 h after pain onset were enrolled in this prospective study. The levels of AT1R-AAs and ETAR ETAR-AAs at the time of hospital admission were measured using an enzyme-linked immunosorbent assay (ELISA) in all patients. The incidence of major adverse cardiovascular events (MACE, defined as a composite of cardiovascular mortality, myocardial re-infarction, and hospitalizations for heart failure) during follow-up was the primary outcome. Autoantibody seropositivity was defined according to ELISA’s kit manufacturer’s instructions (levels > 10 U/mL). Results Two hundred patients with STEMI were enrolled. The baseline characteristics of the patients are shown in Table 1. Of these, 110 (55%) were seronegative for both autoantibodies, 44 (22%) were seropositive for one autoantibody (but not both), and 46 (23%) were seropositive for both autoantibodies. The incidence of MACE over a 14-month median follow-up was higher in patients with double (31%) and single (25%) seropositivity than in seronegative patients (13%, p=0.009 and p=0.06, respectively). Survival free from MACE was significantly different across the different degrees of autoantibody seropositivity (Figure 1). In the multivariable Cox regression analysis, adjusted for the variables that were significant in the univariate analysis (highlighted in bold in Table 1), double seropositivity was independently associated with an increased risk of MACE (hazard ratio 2.386, 95 % CI 1.471-3.864, p<0.001). Conclusions The degree of seropositivity for AT1R-AAs and ETAR-AAs is associated with an increased risk of MACE after STEMI. Our findings provide valuable mechanistic insights into the pathophysiology of STEMI and pave the way for future therapies focused on the improvement of long-term prognosis after acute coronary syndromes.

Bullous Pemphigoid Severity and Levels of Antibodies to BP180 and BP230

The correlation between serum levels of autoantibodies against bullous pemphigoid (BP) antigens 180 (BP180) and 230 (BP230) with BP disease severity is unclear. To investigate the correlation of anti-BP180 and anti-BP230 immunoglobulin G (IgG) antibody levels with BP disease severity. A search was performed of the Cochrane Central Register of Controlled Trials, Embase, and PubMed databases from their respective inception to April 11, 2024. Studies evaluating the correlation between serum levels of anti-BP180 or anti-BP230 IgG measured using enzyme-linked immunosorbent assay (ELISA) and disease severity assessed per the Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) or BP Disease Area Index (BPDAI) were included. No language or geographic restrictions were imposed. Nearly 0.4% of initially identified studies met the selection criteria. One researcher extracted data and another researcher confirmed data. The risk of bias was independently assessed by these researchers using the Quality Assessment of Diagnostic Accuracy Studies 2 tool, with discrepancies resolved by discussion with a third researcher. A random-effects model meta-analysis and a subgroup analysis were conducted based on the ELISA kit manufacturers. Pooled correlation coefficients of antibody levels with ABSIS and BPDAI. In all, 14 studies with 1226 participants were analyzed. The risk of bias of included studies was generally low. The meta-analysis found anti-BP180 autoantibody levels showed moderate correlation with objective BPDAI (r = 0.56; 95% CI, 0.46-0.64) at baseline, strong correlation (r = 0.63; 95% CI, 0.39-0.79) at 3-month follow-up, and moderate correlation (r = 0.53; 95% CI, 0.25-0.72) at 6-month follow-up. Anti-BP180 autoantibody levels also showed moderate correlation (r = 0.52; 95% CI, 0.39-0.62) with ABSIS at baseline, strong correlation (r = 0.62; 95% CI, 0.39-0.79) at 3-month follow-up, and moderate correlation (r = 0.53; 95% CI, 0.25-0.72) at 6-month follow-up. By contrast, anti-BP230 autoantibody levels showed no association with objective BPDAI and ABSIS at diagnosis and follow-up. The subgroup analysis found similar results when using different ELISA kits. The findings of this systematic review and meta-analysis indicated that anti-BP180 autoantibody levels may serve as an adjunctive tool for monitoring BP disease severity and guiding clinical care for patients with BP.

Evaluation of Lactobacillus Sakei Probiotic Supernatant’s Effectiveness in Promoting Apoptosis and Decreasing Breast Cancer MCF-7 Cancerous Cells through Modulation of Bax and Bcl2 Genes

Introduction: Accumulating evidence has revealed that inducing apoptosis is an important strategy to control excessive breast cancer cell proliferation. In this study, the supernatant effect of the probiotic strain of Lactobacillus Sakei on Expression Level of Apoptosis-Related Genes (Bax/Bcl2) in Breast Cancer Cell Line (MCF-7) was investigated. Methods: In the experimental study, MCF-7 breast cancer cells were cultured in DMEM medium with 10% bovine serum. The cells were treated in 1, 2, 3, 4 and 5 mg/ml concentrations of sakei supernatant and incubated at 24, 48 and 72 hours. The MTT assay was used to measure cytotoxicity effect according to kit manufacturer's protocol in all three incubation times. MCF-7 cells with concentration of sakei supernatant at IC50 point (5 mg / ml) to examine the expression of genes Bax and Bcl2 were incubated for 72 hours. Real-Time PCR was performed to analyze the changes in the expression of Bax and Bcl2 genes. Results: The results of this study indicated that bioavailability of MCF-7 cell line reached the lowest value at concentration of 5 mg/ml compared to the control group. In addition, MCF-7 cell line treatment with Lactobacillus sakei supernatant at a dose of 5 mg/ml and 72 h incubation time, showed significantly increased expression of BAX (p-value= 0.0033) and significantly decreased expression of BCL-2 (p-value= 0.0278). Conclusion: The results indicate that Lactobacillus sakei supernatant can reduce the bioavailability of breast cancer cells by inducing apoptosis pathway.

ISOLATION AND ANALYSIS OF NATIVE DNA AND RNA FROM ROOTS OF RUBBER-PRODUCING SPECIES SCORZONERA TAU-SAGHYZ LIPSCH. ET G.G. BOSSE

Scorzonera tau-saghyz Lipsch. et G.G.Bosse, is endemic species, growing in South Kazakhstan and alternative (to Hevea brasilians) source of natural rubber, related to the Asteraceae Family. Natural rubber (NR) is an indispensable raw material for the production of different products for medicine, automotive and aviation industries. The introduction of tau-saghyz into the culture will make it possible to obtain NC not only in Kazakhstan, but also on the territory of the countries of the Northern latitudes. At the growing phases of ontogenesis of tau-saghyz (April-June) latex is actively synthesized in the leaves and transported to the roots. In our experiments it was necessary to conduct molecular genetic studies of the selected forms of tau-saghyz with high rubber concentration. In this regard, it became necessary to isolate and analyze native DNA and RNA from dry roots. Samples with the highest rubber content were used for DNA extraction. The main problem in the isolation of DNA from the dry roots of tau-saghyz is the significant content of natural rubber. In this research the following commercial kits were used: 1) DNeasy Plant Mini Kit (Qiagene); 2) GenElute Plant Genomic DNA Miniprep Kit (Sigma); 3) NucleoSpin® Plant II / Midi / Maxi kit (Macherey-Nagel). Further DNA isolation was carried out in accordance with the kit manufacturer's protocol. Quantitative DNA analysis was performed using both UV spectrophotometry (Nanodrop) and fluorometric analysis (Qubit) method. DNA extracted by the 2 kit (Sigma) was further purified by precipitation with ethanol and demonstrates satisfactory quality. Functional analysis of isolated DNA samples was performed both by restriction analysis and by DNA amplification. It is worth to mention that for reaction primers that are characteristic of gene AACT (ACETOACETYL-COA TRANSFERASE) Cucumis melo were used. This enzyme catalyzes the first reaction of biosynthesis in the mevalonate pathway, which the final product is IDP (isopentenyl diphosphate). The cis-prenyltransferase (CPT) enzyme catalyzes the biosynthesis of rubber by the sequential addition of IDP to the terminal group of the initiating allylic pyrophosphate (APP). As a result of PCR analysis a putative 1200 bp AAST gene for tau-saghyz was identified. In this study two methods of RNA isolation were chosen: 1. A method widely used for plants of the Cucurbitaceae family; 2. A method approved for plants with a high content of polyphenols. The RNA isolation was carried out as described in the relevant protocols with minor modifications (Verwoerd TC, Dekker BMM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17:2362). The final precipitates were dissolved in 50 µl of water pretreated with diethylpyrocarbonate. Quantitative analysis of RNA was carried out using UV spectrophotometry (Nanodrop). Functional analysis of extracted RNA samples was conducted by cDNA synthesis with usage of reverse transcriptase Superscript IV with the following amplification in PCR. In this reaction the primers specific for CPT gene of Taraxacum kok-saghyz were used. The prospective CPT gene was excised from the gel, purified and cloned into the pJET vector. In the further research it is planned to perform its sequencing and comparison with the CPT gene of Taraxacum kok-saghyz and Hevea brasiliensis.

Formal and Legal Analysis of the Issues Related to the Placing on the Market and using Fire Pumps and Pump Kits

Aim: The aim of this article is to present the existing formal and legal conditions related to the placing on the market and using in the national market of pumps and pump kits – both fire pumps and those intended for firefighting water supply systems. The paper also discusses other selected regulations relating to the product group in question. In addition, an issue not arising from the construction regulations, but equally important in terms of ensuring an adequate level of safety in buildings, was raised, i.e. the admittance of control panels for firefighting equipment (in this case pumps), which can form an integral part of the pump kits in question. The results of the analysis can also be applied to other selected products in the area of fire protection and the requirements that these products must meet before they can be placed on the market and put into use. Introduction: Fire water systems are among the most commonly used fire extinguishing systems in buildings. Despite their popularity, quite often investors, designers and users of buildings, as well as the manufacturers of pump kits themselves, have doubts about the requirements for placing these products on the market. The authors’ discussion of the current requirements is intended to consolidate knowledge in this area, clarify the marketing of construction products and resolve possible doubts. Methodology: The publication uses the results of the authors’ own analysis on the basis of the interpretation of the legislation, as well as available normative and literature sources. Conclusions: The analysis shows that fire pumps and pump kits are covered by the obligation to carry out national assessment and verification of constancy of performance in system 1 and to draw up a national declaration of performance as well as to mark the product with a construction mark (before placing it on the market or making it available on the national market). Moreover, from 1 January 2021, the obligation under consideration was extended to pumps for fire water supply systems, and from 1 July 2022 also to pump kits for fire water supply systems. Furthermore, it was pointed out that the sub-assembly acting as the control panel for the firefighting equipment, insofar as it is present in the pump kits in question, is also subject to admittance Keywords: fire pumps, pumps for firefighting water supply systems, “B” construction mark, national technical assessment

Glia Maturation Factor Beta: A Novel Neuro-Impairment Prediction Factor in Toxoplasmosis.

Toxoplasma gondii, a neurotropic protozoan, infects up one to third of the world population. The parasite can invade a wide variety of nucleated cells but preferably glial cells. Glia maturation factor β (GMFβ), a 17KD protein expressed at high levels in the central nervous system is predominantly related to neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Multiple sclerosis. We aimed to determine the expression level of GMFβ and its relation to other pro-inflammatory factors (IL33, SDF1, and CCL2) on T. gondii infected human neuroblastoma cell line. The human neuroblastoma (SK_NMC C535) cell line was infected by 5×106 (1:1 ratio). The supernatant was collected after cell lysis and centrifugation. Total RNA was extracted using the Yekta Tajhiz RNA extraction kit. cDNA was synthesized based on RevertAid First Strand cDNA Synthesis Kit manufacturer's protocol (Parstous, cDNA synthesis kit, Iran). The specificity of each primer pair (GMFβ, IL33, SDF1, and CCL2) was provided by NCBI BLAST. Gene expression level was measured using Real-Time PCR. All experiments were conducted at the Hamadan University of Medical Sciences, western Iran in 2022. The GMFβ increased significantly up to 1.35-fold (P=0.007). The increase in GMFβ expression in neuroblastoma cells was consistent with the increase in pro-inflammatory factors (CCL2 (0.47), IL33 (0.152) and, SDF1 (1.33)). GMFβ upregulation can be a novel indicator of the destruction of nerve cells.

A novel derivatization method for the determination of ethyl carbamate in spirits by liquid chromatography with spectrophotometric detection

A novel derivatization method for the ethyl carbamate (EC) determination is introduced, representing a significantly more promising approach to the only previously described and time-demanding EC derivatization with the expensive 9–xanthydrol agent. The presented derivatization procedure utilizes a 4–(dimethylamino)benzaldehyde (p-DMAB) reagent to form a derivative with a recovery of 99.4 %. The quantitative EC reaction with p-DMAB proceeds within five minutes without any considerable by-products and the resulting derivative was confirmed by mass spectrometry. The derivatization procedure led to the development of a new chromatographic method with spectrophotometric detection at 420 nm for EC determination in spirits. The achieved quantification limit of 582.5 µg L–1 complies with the recommendation of the European Commission from 2010, demonstrating its practical use in food safety control. Within the optimization of detection, the electrochemical behaviour was included to reveal the possibility of amperometric detection. Compared to the method used so far, the proposed approach significantly reduces the cost and time required to screen this potential carcinogen and could lead to the manufacture of commercial colorimetric assay kits detecting hazardous EC concentration levels.

High Diagnostic Accuracy of Thyroid-Stimulating Hormone (TSH) Receptor Antibodies in Distinguishing Graves' Disease and Subacute Thyrotoxicosis in the Indian Population.

Thyrotoxicosis is a common clinical condition encountered in endocrine practice. Graves' disease and subacute thyroiditis are the two common causes of thyrotoxicosis and often have overlapping clinical and biochemical features. 99mTc thyroid scintigraphy is the most commonly used confirmatory test to differentiate the two conditions but is not available in the majority of the second-tier cities of India. However, obtaining thyroid stimulating hormone (TSH) receptor antibodies (TSHrAb), another accurate test to differentiate the two conditions, in second-tier cities by outsourcing to labs in major cities is a feasible option nowadays. However, the data on the performance of TSHrAb to differentiate the two conditions in Indian patients is limited. Hence, we have evaluated the diagnostic accuracy of TSHrAb in the Indian population to differentiate Graves' disease and subacute thyroiditis. This prospective studywas conducted on 115 consecutive newly diagnosed thyrotoxicosis patients presenting to the Department of Endocrinology at a tertiary health care centre in India. Clinical parameters like throat pain, duration of symptoms, and grade of goitre were noted. Measurement of total tri-iodothyronine (TT3), total thyroxine (TT4), TSH, TSHrAb, and 99mTc thyroid scintigraphy were performed in all participants. All participants were followed up at least for six months after the recruitment. Increased tracer uptake (>4%) and/or increased thyroid to parotid trace uptake ratio (>2.5) were used to diagnose Graves' disease. Eighty-one and 34 patients were diagnosed with Graves' disease and subacute thyroiditis, respectively. TT3/TT4 ratio had low diagnostic accuracy (area under the curve (AUC): 0.6, best cut-off: 15.6, sensitivity: 53.1%, specificity: 79.4%). TSHrAb had the best AUC (0.9) to distinguish Graves' disease from subacute thyroiditis (cut-off: 2.0 IU/L, sensitivity: 97.5%, specificity: 100%). In contrast, the kit manufacturer's reference range (1.75 IU/L) was slightly more sensitive (98.8%), but less specific (94%). The TT3/TT4 ratio is not a good test to differentiate Graves' disease and subacute thyroiditis. TSHrAb is accurate in distinguishing Graves' disease from subacute thyroiditis and a level of 2.0 may be a more accurate cut-off to differentiate the two conditions in the Indian population.

Investigation of proapoptotic and cytotoxic effects of 2-aminobenzothiazole on human laryngeal carcinoma cells.

In the present study, we investigated the effects of 2-aminobenzothiazole application on human laryngeal carcinoma cells. Human larynx epidermoid carcinoma (HEp-2) (ATCC® CCL-23™) cells were purchased from American Type Culture Collection (ATCC, USA). Human larynx epidermoid carcinoma HEp-2 cells were cultured in complete Dulbecco's Modified Eagle's Medium (DMEM) supplemented with fetal bovine serum (FBS) (10%) and penicillin/streptomycin (1%) in a CO2 (5%) incubator under standard cell culture conditions. 2-aminobenzothiazole was prepared, and further dilutions ranging from 3.13 to 100 μM were prepared in fresh culture DMEM. HEp-2 cells on 96 well plates were incubated with the prepared dilutions of 2-aminobenzothiazole for 24, 48, and 72 hours. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test performed cytotoxicity evaluation and viability percentages. The annexin-V staining technique detected 2-aminobenzothiazole-triggered apoptosis of HEp-2 cells. The activated caspases 3/7 on HEp-2 cells after 2-aminobenzothiazole exposure were evaluated with flow cytometric analysis. The membrane potential changing of HEp-2 cells was measured following the Muse™ MitoPotential kit manufacturer instructions. MTT cytotoxicity test results showed that the viability of human laryngeal carcinoma cells decreased with an increase in the application of 2-aminobenzothiazole for 24 hours. The highest growth inhibition by 2-aminobenzothiazole for short-term application of 24 hours was detected at the highest concentration of 2-aminobenzothiazole (100 µM). The results underline that the cytotoxic effect of 2-aminobenzothiazole is dose-dependent. Cytotoxicity test results for an application time of 48 hours showed that the cytotoxicity of 2-aminobenzothiazole is dose-dependent on HEp-2 cells. The required dose of 2-aminobenzothiazole to decrease the cell viability to 50 percent has been 9-fold augmented. Annexin-V findings showed that after exposure to IC50 concentration of 2-aminobenzothiazole for 24 hours, HEp-2 cells underwent the early apoptotic stage (25.99%) and late apoptotic (16.69%), whereas 56.93% of the treated cells were alive. Only 0.39% of 2-aminobenzothiazole treated cells were necrotic. All study results showed that 2-aminobenzothiazole triggered apoptosis on HEp-2 cells with a percentage of total apoptotic cells 42.62 compared to untreated HEp-2 cells. Caspase 3/7 activation results showed that only 0.65% of control HEp-2 cells were with activated caspase 3/7, and 99.35% live cells. The analysis data from the Muse cell analyzer revealed that the percentage of cells with intact mitochondrial membranes was 21.30 after 2-aminobenzothiazole application, and 79.9% were cells with depolarized mitochondrial membranes. It has been understood that the depolarization of the inner mitochondrial membrane has been considered a dysfunction in mitochondria as a sign of apoptosis and drug toxicity. Based on all study findings, 2-aminobenzothiazole has cytotoxicity on human laryngeal carcinoma cells in a dose and time-dependent manner. That means that it decreased viability via inducing caspase-dependent apoptosis. Consequently, it was concluded that 2-aminobenzothiazole has good potential to lead to cytotoxicity and apoptosis on human laryngeal carcinoma cells and, after deeper in vitro and in vivo investigations, can be a good candidate for designing anticancer drugs with high efficiency.

Serum Thyroglobulin (Tg) assay by immunoradiometric methods (IRMA): comparison of concentrations obtained with Cisbio Bioassays vs Beckman Coulter kit in patients with differentiated thyroid cancer.

Introduction: In clinical practice, Thyroglobulin is a tumor marker for the post-operative follow-up of differentiated thyroid cancer, provided that any thyroid residual remains are eliminated beforehand, either by simple total thyroidectomy, or most often supplemented by isotopic totalization with Iode131. The aim of this study were to compare serum Tg values measured with 02 kit manufacturers of immunoradiometric type (IRMA) and to evaluate their cost/effectiveness ratio. Methods: We included patients followed for differentiated thyroid cancer operated and irradiated with Iodine 131 at the nuclear medicine department of Tlemcen University Hospital. Their serum Thyroglobulin was dosed in the laboratory of the department with the Tg-IRMA kit of Cisbio Bioassays taken in this study as «Gold standard» and the Tg-IRMA kit of Beckman Coulter. Results: Ninety-nine (99) serums of patients with CDT were measured for Thyroglobulin. The correlation of the Thyroglobulin values obtained with the 02 kits is positive and significant (r= 0.98; p < 0.01). The equation of the regression line of Passing- Bablok is of type y = 0.7 x + 0 (with Y = Tg-IRMA Cisbio Bioassays and X = Tg-IRMA Beckman Coulter). The difference in the averages analyzed with the Bland-Altman diagram is = 0 (p > 0.05), indicating that there is no difference in the mean of Tg between the 02 kits. The analysis by ROC curve of the Tg-IRMA Beckman Coulter kit Thyroglobulin values at the threshold of 0.7 and 1 ng/mL finds respectively a sensitivity and a specificity of 100% and 93% with an area under the curve = 0.99. Conclusion: There is a strong correlation between Thyroglobulin concentrations obtained with the 02 kits, so a budget reduction in the management of our patients with diffrentiated thyroid cancer is possible, in favor of the Tg-IRMA kit of Beckman Coulter with a good cost/ efficiency ratio.

A-051 Reference Intervals for Serum T4 and FT4 among Pregnant Women linked to the Mãe Curitibana Program

Abstract Background Thyroid diseases are frequent in women of childbearing age and can have negative maternal and fetal repercussions when not identified and properly treated. Physiological changes during pregnancy cause changes in thyroid function tests, which is why they must be carefully interpreted and, whenever possible, based on local and specific reference values for pregnant women. Determining the serum level of total (T4T) or free (FT4) T4 is fundamental for defining the diagnosis of thyroid dysfunctions between clinical (TSH and T4 altered) or subclinical (only TSH altered). This definition modifies the treatment recommendations for thyroid disorders in various published guidelines. Objectives To determine the reference intervals for FT4 and T4T of pregnant women, in the first trimester, residing in the city of Curitiba and followed by the municipal public health system. Methods Longitudinal study with a sample of 225 pregnant women aged over 18 years, linked to prenatal care in the public health system (SUS) of Curitiba city, with up to 14 weeks of gestation. Pregnant women were selected proportionally to each health district in the city, to represent the entire population of the city served by the SUS. Pregnant women who had TSH outside the reference range, positive ATPO, family history of thyroid disease, use of antithyroid drugs, use of medications that could interfere with the measurement of the markers under study were excluded. The studied population had a median urinary iodine concentration considered adequate for pregnant women (158 mcg/L). Serum FT4 and T4T were measured by chemiluminescent immunoassay on Atellica IM Analyzer. Continuous variables were expressed as mean and standard deviation and percentiles were determined and variations between 2.5 and 97.5 percentiles were considered as the normal reference interval. The results were analyzed and compared with the reference interval provided by the manufacturer for non-pregnant women. Results Of the 225 pregnant women (with mean ± standard deviation of gestational age 8 weeks ± 2.3; with mean ± standard deviation of age 27 years ± 5.9) the FT4 values of the pregnant women ranged between 0.83 and 1.68 ng/dL, with mean ± standard deviation of 1.23 ± 0.15. The 2.5th and 97.5th percentiles were 0.94 and 1.64 ng/dL. Compared with the kit manufacturer’s reference values for non-pregnant women (0.95–1.47 ng/dl), we would have 08 pregnant women with a high FT4 result. The T4T values of the pregnant women varied between 9.2 and 12.2 ng/dL, with a mean ± standard deviation of 10.9 ± 2.48. The 2.5th and 97.5th percentiles were 9.2 and 12.2 ng/dL. Compared with the kit manufacturer’s reference values for non-pregnant women (4.87 and 11.72 ng/dL), we would have 25 pregnant women with high T4T results. Conclusion The FT4 and T4T reference intervals of first trimester pregnant women are different from the non-pregnant population, however this difference is less relevant in FT4. In T4T, the lower limit of normal in pregnant women is much higher than in the normal population, which makes it less sensitive for the definition of clinical hypothyroidism in first trimester pregnant women.

Fasting plasma insulin in normal-weight apparently healthy adults in a Nigerian population: Reference intervals and influence of age, gender and anthropometric variables

Introduction: In Nigeria, there is paucity of data regarding indigenously established reference intervals for fasting plasma or serum insulin concentrations in the local populations. This study was designed to establish the reference interval of fasting plasma insulin (FPI) among apparently healthy young and middle-aged adults in a Nigerian population. The influence of age, gender, and anthropometric variables on FPI concentration were examined. Materials and Methods: The study was a cross-sectional descriptive study involving 210 reference individuals aged 18 to 64 years. Physical, anthropometric and biochemical variables were measured including FPI and fasting plasma glucose (FPG). The reference interval for FPI was determined using the non-parametric percentile method. Correlation studies between FPI and age, anthropometric indices, and HOMA-IR were carried out. Results: The study involved a total of 210 healthy normal-weight non-diabetic adults consisting of 110 males (52.4%) and 100 females (47.6%) as reference individuals. Reference intervals for FPI for the male, female, and total study participants were 0.1 –11.1mIU/L, 1.3 –13.2mIU/L and 0.1 – 13.03mIU/L respectively. There were statistically significant positive correlations between FPI and age (r = 0.001), WC (r = 0.0.302, p = 0.0001), WHR (r = 0.220, p = 0.0001), SBP (r = 0.137, p = 0.047) FPG (r = 0.165, p = 0.017) and HOMA-IR (r = 0.985, p = 0.0001). Conclusion: The reference interval of FPI using the Biointecho human insulin ELISA kit is 0.1–13.0mIU/L. This is not much at variance with 0.7–9.0mIU/L quoted by the kit manufacturer, but valued widely with that of other commercially available insulin assay kits from different manufacturers. Similar to reports of similar studies, FPI correlated positively with FPG, BMI. WC, BP and age.

Department of Microbiology, Collage of Veterinary Medicine, University of Diyal, Iraq

This study was done at the college of Veterinary Medicine, University of Diyala, from September 2021 to April 2022 to assess the seroprevalence and molecular detection of avian influenza virus (AIV) H5N8 in layer chickens. All samples came from 8 commercial flocks of layer chickens. Clinical sings included, severe respiratory infection with enlargement of the head, neck, and prominent edema of the face, followed by excessive lacrimation, sinusitis, severe rales, and cyanosis in all parts of unfeather skin predominantly the wattles comb, and the legs. In this regard, ELISA was used as a serological test and RT-PCR as a molecular detection technique. Accordingly, (364) serum samples from 8 probable AIV (H5N8) infected layer farms were collected and subjected to ELISA test using commercial kit that specific for IgG antibodies to AIV H5N8. The results showed a significant increase of IgG antibodies in such serum samples at days 70 and 200 of age. According to the instruction manual of ELISA kit manufacturer and to mean titers of layers of present study, all farms are infected with AIVH5N8 strain. However, at the age of 200 days, 48 postmortem tissue samples including trachea, lung and liver of affected birds were collected from clinically and sub-clinically infected flocks from all farms. These samples were processed, RNA extracted and submitted to RT-PCR using specific primers for H5N8 strain. The results showed that 32 out of the 48 tissue samples (66.6%) tested positive for H5N8.The resulted Amplicon (320bp) was commercially sequenced and analyzed. The sequencing of the local AIV H5N8 revealed a 99% sequence identity with the reference sequences. The detected strain was registered in GenBank data (NCBI) under acc.number ON247929.1. Phylogenetic tree for locally detected virus in comparison to data from of NCBI was created and showed that the investigated S1(ON247929.1, AIV (A/laying hens/Iraq/(H5N8) segment 4 hemagglutinin (HA) gene, partial cds, local sample is closely related to reference isolates from the NCBI acc. no. of MW961428.1, MW961444.1, MW961476.1, MW961436.,1 and MW961484.1. These strains of the Influenza H5 virus have been deposited from Nigeria.

Biodiversity of Drosophila sp. from the Natural Environment based on the Cytochrome Oxidase subunit 1 Gene

Research on insect phylogenetics is intricated by their similar morphology and significant genetic diversity. The cytochrome oxidase subunit 1 (CO1) gene is the most widely utilized mitochondrial DNA gene in the identification and study of animal molecular biodiversity. This study aims to identify and reconstruct the phylogeny of fruit flies from North Sulawesi using the cytochrome oxidase subunit 1 (CO1) gene. Fruit flies were obtained from 5 (five) areas in North Sulawesi, namely Siau (L1), North Minahasa (L2), Minahasa (L3), Southeast Minahasa (L4), and Bolaang Mongondow (L5). Fruit fly imago limbs were used as a tissue source for genomic DNA extraction. Genomic DNA extraction was carried out using the Quick-DNA™ Miniprep Kit manufacture protocol. The CO1 gene amplification was carried out by the PCR method, and the visualization of the amplicons was carried out by the 1.5% gel electrophoresis method. Nucleotide sequencing used a sequencing service at First BASE Singapore with a bidirectional sequencing method. CO1 gene amplification of each sample was visualized at 690 bp to 702 bp length. After analyzing the CO1 gene concession area using the MEGA XI program, it is found that Drosophila at L1 has 702 bp, L2 has 703 bp, L3 has 698 bp, L4 has 700 bp, and L5 has 697 bp. Based on alignment analysis using the BLAST method, it is found that the L1 fruit fly has a similarity rate of 99.29% (E=0.0) to Drosophila parapallidosa [MK659836.1]. The L2 fruit fly also has a similarity rate of 96.86% with Drosophila parapallidosa [MK659836.1]. The L3 fruit fly has a similarity level of 94.94% with Drosophila parapallidosa [MK659836.1]. The L4 fruit fly has a similarity rate of 94.43% with Drosophila parapallidosa [MK659836.1]. However, the L5 fruit fly shows a similarity rate of 96.86% with Drosophila rubida [EU493593.1]. The reconstruction results with the MEGA XI program using the Minimum Evolution model obtain two monophyletic groups where the fruit fly in Bolaang Mongondow is in a monophyletic group different from other fruit flies. The results of this study prove the variation in fruit fly species in North Sulawesi based on the identification of the CO1 gene.

Significance of 3D printing for a sustainable environment

The additive manufacturing, also known as 3D printing, allows for a complete control over the entire manufacturing process that can be tuned to any application. In recent years, it has been tested for its role in various areas like environmental contaminant monitoring, providing solutions for the energy generation and storage and healthcare. To encourage sustainable detection platforms for the pollutants, the sensing electrodes have been reported to be 3D printed because of enhanced electrochemical properties coming from the high surface area of the printed materials. In general, the conventional methods of electrode preparation are time consuming, expensive, and mostly not adaptable. 3D printing however, negates all these challenges. Similarly, in the energy generation and storage field, the rapid and lightweight materials used during 3D printing make them viable and suitable alternative. Since 3D printing is a bottom-up method for the fabrication, the amount of raw material consumed can be tuned and the by-products or wastage can be minimized. Apart from these demanding areas, additive manufacturing, in last 2 years which witnessed epidemic outspread, has supported health sector immensely in fight against COVID. 3D printing allowed the rapid manufacturing of COVID-19 detection kits and helped to maintain the COVID-19 safeguards in place. Sample collection swabs, respirators, and other components of the personal protective equipment kits were among many products developed using 3D printing during the pandemic. Keeping these things in mind, this review encapsulates the use of 3 D printing for energy application, detection of water and biological contaminants, and as safeguard tool during COVID pandemic.

Anything But a Biopsy: The Quest for Noninvasive Alternatives in Heart Transplantation.

Anything But a Biopsy: The Quest for Noninvasive Alternatives in Heart Transplantation.

FREQUENCY OF TRANSFUSION-TRANSMISSIBLE INFECTIOUS DISEASES AMONG BLOOD DONORS AT AKHTER SAEED TRUST HOSPITAL LAHORE, PAKISTAN

Background: Blood transfusion is an essential lifesaving treatment. The unsafe blood transfusion practices are one of the reasons of spreading transfusion-transmissible infections among individuals. It is necessary to screen the all donated blood units for HBV, HCV, HIV, VDRL, and MP.
 Aims: This aim of this study was to evaluate the burden of transfusion-transmissible infectious diseases caused by blood transfusions among donors.
 Materials & Methodology: It was a prospective observational study which included the 9114 blood donors who attended the Akhtar Saeed Trust Hospital Lahore from January 2020 to September 2022. The informed consent was taken from all blood donors. 3-5 ml of venous sample was drawn by using aseptic technique from blood donors on enrollment. Screening of blood donors was done by Chemiluminescence immunoassay (Maglumi-800) for HBV, HCV, HIV and VDRL. The MP was confirmed by peripheral blood picture on slide. According to the automation kit manufacturer’s guidelines, cut-off value for reporting positive results of HBV, HCV, HIV and VDRL was calculated. Data was analyzed by using SPSS version 25.0.
 Results: The mean age was 27.79 + 12.1 years. The numbers of volunteer and replacement donors were 961 (10.54%) and 8153 (89.45%) respectively. The total numbers of positive donors for transfusion transmissible infectious diseases were 591/9114 (6.49%). The seropositivity was found to be 170/9114 (1.86%) for HBV, 324/9114 (3.55%) for HCV, 33/9114 (0.36%) for HIV, 64/9114 (0.70%) for VDRL, and 0/9114 (0.0%) for MP respectively.
 Conclusion: There is currently a low risk of transfusion transmissible infectious diseases but the availability of safe blood is contingent on screening tests and appropriate donor selection.
 

Evaluation of Vitamin D level in thalassemia patients: The experience of a single center

BACKGROUND: Beta-thalassemia, a hereditary blood disease transmitted through families, has become increasingly relevant with rising life expectancies, leading to bone disease being a significant cause of morbidity. Among the symptoms observed in these patients, bone pain and back pain are frequently reported. Vitamin D is believed to play a crucial role in reducing these symptoms. AIM: The objective of this study was to assess the Vitamin D levels in thalassemic patients and investigate potential correlations with other factors. PATIENTS AND METHODS: A cross-sectional study was conducted, involving a random selection of 48 patients with beta-thalassemia (major and intermediate types) aged 7 years and above. The patients were registered at the Hereditary Blood Disease Center in Wasit province, located in the South of Iraq, during the period from January to May 2022. Demographic data, including age, sex, address, diagnosis, type of chelation therapy, and frequency of blood transfusions, were collected from patients’ files. Biochemical data, such as mean hemoglobin, mean serum ferritin, mean serum calcium, and Vitamin D levels at the time of the study, were also recorded. Vitamin D levels below 30 ng/ml were considered low level or deficiency, whereas 30 ng/ml and above were considered normal, as indicated by the kit manufacturer. Furthermore, the height, weight, and body mass index were evaluated in the studied patients with their written consent. SPSS version (23) was employed for data management and statistical analysis, utilizing a significant P = 0.05 and the Pearson’s correlation. RESULTS: The study revealed that 42 patients (87.5%) had low Vitamin D levels (below 30 ng/ml). The mean Vitamin D level was 18.23 ng/ml, with a maximum reading of 45 ng/ml. It was observed that Vitamin D deficiency was more prevalent in younger patients and those with higher ferritin levels, although the differences were not statistically significant. CONCLUSION: Vitamin D deficiency is common in patients with B-thalassemia, as indicated by this study.

CODA: a combo-Seq data analysis workflow.

The analysis of the combined mRNA and miRNA content of a biological sample can be of interest for answering several research questions, like biomarkers discovery, or mRNA-miRNA interactions. However, the process is costly and time-consuming, separate libraries need to be prepared and sequenced on different flowcells. Combo-Seq is a library prep kit that allows us to prepare combined mRNA-miRNA libraries starting from very low total RNA. To date, no dedicated bioinformatics method exists for the processing of Combo-Seq data. In this paper, we describe CODA (Combo-seq Data Analysis), a workflow specifically developed for the processing of Combo-Seq data that employs existing free-to-use tools. We compare CODA with exceRpt, the pipeline suggested by the kit manufacturer for this purpose. We also evaluate how Combo-Seq libraries analysed with CODA perform compared with conventional poly(A) and small RNA libraries prepared from the same samples. We show that using CODA more successfully trimmed reads are recovered compared with exceRpt, and the difference is more dramatic with short sequencing reads. We demonstrate how Combo-Seq identifies as many genes and fewer miRNAs compared to the standard libraries, and how miRNA validation favours conventional small RNA libraries over Combo-Seq. The CODA code is available at https://github.com/marta-nazzari/CODA.

Implementation of Public Health Genomics in Africa: Lessons from the COVID-19 pandemic, challenges, and recommendations.

Public Health Genomics (PHG) is a relatively new field. The wide application of genomic technologies played a pivotal role in elucidating the full genomic sequence of the SARS-CoV-2 virus. This breakthrough proved to be the starting point in the manufacture of diagnostic kits and the subsequent making of vaccines. Beyond the COVID-19 pandemic, many African countries can take advantage of the various investments in genomic technologies to introduce and intensify the use of genomics for public health gain. Public Health Genomics effectively monitors, prevents, and manages non-communicable and infectious diseases. However, there are several challenges to implementing PHG in Africa. In this perspective article, we discuss the utilization of PHG during the COVID-19 pandemic, the lessons learned from using PHG to manage and contain the COVID-19 pandemic, as well as potential challenges Africa may face when putting PHG into practice compared to challenges of other regions. We also discuss our recommendations for overcoming these challenges.