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Introduction: District Sheikhupura encountered its first dengue outbreak in 2014 but lacked serological evidence and reports of risk factors associated with it. To assess this, a hospital-based study was conducted from January 2014 to December 2017. Methodology: Blood from 333 participants was collected, the serum obtained was tested for IgG and IgM antibodies against DENV using a commercially available ELISA kit. Results: The results showed that out of all (n= 333) samples tested, 120 were turned up positive for DENV, making an overall prevalence of 36%. Of the 120 confirmed cases, 55% (n = 66) were recorded in 2014, 10% (n = 12) in 2015, 27.5% (n = 33) in 2016, and 7.5% (n = 9) in 2017. It was found that 68.3% (n = 82) were male and 31.7% (n = 38) were female, with 61% (n = 74) patients aged between 11-30 years. The highest prevalence of infection, 94.2% (n = 113), was noted after the rainy season. During the study, the highest number of cases appeared in Ferozewala Tehsil. The factors age, gender, and season were found statistically significant with the prevalence of infection (p < 0.05). Conclusions: The study is the first report on the detection of dengue in the Sheikhupura district. The survey anticipated its geographical expansion, determined associated risk factors, and suggests active disease surveillance in the area.

Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of

Objective Patients who develop interferon-gamma autoantibodies (IFN-ɤ autoAbs) in adult-onset immunodeficiency (AOID) syndrome are more likely to develop opportunistic and recurrent intracellular infections. The assay to detect IFN-ɤ autoAbs is essential for the diagnosis and therapeutic monitoring of AOID syndrome. Therefore, this study applied the QuantiFERON assay for the detection of IFN-ɤ autoAbs. Methods Serum from patients with AOID syndrome (n = 19) and serum from healthy patients (n = 20) was collected and applied using 2 neutralizing platforms of enzyme-linked immunosorbent assay (ELISA) kits (the BD ELISA and the QuantiFERON ELISA) for IFN-ɤ autoAbs detection. Results The pooled serum from patients with AOID syndrome showed >50% inhibition at 1:5000 dilution (positive), whereas the pooled serum from healthy patients showed 1:5,000,000 according to the neutralizing QuantiFERON ELISA. Conclusion The QuantiFERON ELISA kit could be applied for the detection of IFN-ɤ autoAbs for the diagnosis and therapeutic monitoring of AOID syndrome.

Vaccination was the main approach to control spreading of FMD disease in Egypt. This study was carried out for evaluation of vaccination by governmental and prevailing local vaccines in cattle sera in menofia governorate during the period from October 2018 to May 2019. A total 130 serum samples were collected to evaluate the antibodies against FMDV. 30 samples were used as control, 50 samples collected from animals received local governmental vaccine and 50 samples from animals received local prevailing vaccine. 49 out of 100 serum samples (37.7%) were positive and 81 out of 130 serum samples (62.3%) were negative for NSP. Results were (0.0), 3(3.7%) pre-vaccination and 27(33.33%), 18(22.22%) 1st MPV- 25(30.86%), 18(22.22%) 2nd MPV-15(18.52%), 13(16.05%) 3rd MPV and (0.0), 9(11.11%) 4th MPV for serotype A. For serotype (O): 9(11.1%), 10(12.4%) pre-vaccination- 36(44.44%), 28(34.56%) 1st MPV-33(40.74%), 27 (33.33%) 2nd MPV-24(29.63%), 20(24.7%) 3rd MPV-14(17.3%), 12(14.80%) 4th MPV. For serotype (SAT2): 4(4.9%), 24(4.93%) pre-vaccination-30 (37.03%), 21(25.93%) 1st MPV-27(33.33%), 21(25.93%) 2nd MPV- 18 (22.22%), 14(17.3%) 3rd MPV-3 (3.71%), 4(4.93%) 4th MPV. In Conclusion. FMD controlling requires vaccines that will supply disparity of infected from vaccinated animals.

Background: Echinococcosis caused by Echinococcus granulosus is a zoonotic disease of public health significance, but there have been few studies of the infection in dogs in Nigeria. This study aimed to establish the seroprevalence of E. granulosus in dogs in four Local Government Area of Plateau State, Nigeria.Methods: A total of 179 dog sera were examined for the presence of E. granulosus antibody using an ELISA kit between May to October 2017.Results: Eleven of the sera (6.1%) were seropositive with a prevalence of 7.4% and 2.3% for dogs less than 2 years and those above 2 years of age respectively. More male dogs (7.5%) than females (4.1%) were seropositive. Seroprevalence was associated with 2 years dogs (OR=3.33, p=0.46) and male than female dogs (OR=1.90, p=0.53) but association were not significant. Dogs from Bassa had the highest seroprevalence (15.2%), followed by Mangu (5.9%), Kanke (4.8%) and Jos South (2.0%). Odds ratio showed an association which was not significant between Bassa (OR=3.3), Kanke (OR=2.50).Conclusion: This study shows that E. granulosus infection is present in dogs in some parts of Plateau State. More studies should be done to ascertain echinococcus infection in dogs in Nigeria to aid the formulation of control programmes to forestall its public health impacts. Annals of Medical Laboratory Science (2021) 1(2), 9 - 17

Tumour protein 53 (p53) plays an important role in the instruction of the cell cycle. In a variety of transformed cell lines, tumour protein is expressed in high amounts, and it is believed to contribute to transformation and malignancy. This research aimed to detect the anti-p53 antibodies in sera of patients with various malignant tumours and to evaluate the sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA). A case-control study was conducted on samples from 49 patients with various types of malignant tumours at Sultanah Bahiyah Hospital, Alor Setar, Kedah, Malaysia, and 32 healthy control cases with non‐malignant disease collected from Universiti Sains Malaysia clinic, Penang, Malaysia. The antibodies against p53 protein in the serum samples were analysed using the commercial ELISA kit, Calbiochem® p53- ELISAPLUS. The results showed that the rate of anti-p53 antibodies in patients with various malignant tumours was 13 out of 49 (26.5 %), compared with only 2 out of 32 (6.25%) in healthy controls (p < 0.001). The sensitivity of this kit reached 28.6% and the specificity was 93.8%. In conclusion, these results suggest that the anti-p53 antibodies can be detected in different sera of malignant tumour patients and the ELISA kit is highly specific; nevertheless, its discrimination power is not perfect because of its low sensitivity to determine the anti-p53 antibodies.

This study evaluated the diagnostic role of cerebrospinal fluid leucine-rich alpha-2 glycoprotein (CSF LRG) concentration in children with acute bacterial meningitis, and its role in differentiation from aseptic meningitis. CSF LRG concentration was measured by ELISA Kit of 50 children with bacterial meningitis, 16 aseptic meningitis, and 20 children with normal CSF; control. CSF LRG was significantly elevated (p

This study was designed to detect the persistent infection with bovine viral diarrhea virus (PI- BVDV) among 114 bulls in Aswan Governorate. Sera samples were examined by ELISA and real time polymerase chain reaction (PCR) to detect BVDV antibodies and BVDV nucleic acid, respectively. The percent of infection was 23.7% (27/ 114) and 76.3% (87/ 114) were negative by ELISA technique, and 3.4% (3/ 87) for BVDV gene detection by real time PCR. Sensitivity of 89%and specificity of 96 % was reported using the commercially available ELISA kit for samples obtained from PI animals.

Mycoplasma gallisepticum (M. gallisepticum) is the primary agent of chronic respiratory disease causing important economic losses in the poultry industry. Compared to antibodies, aptamers used to diagnose M. gallisepticum have many advantages, such as being chemically, animal-free produced and easily modifiable without affecting their affinity. Herein, a single-stranded DNA (ssDNA) aptamer Apt-236 which can specifically bind to PvpA protein of M. gallisepticum with a Kd of 1.30 ± 0.18 nM was selected successfully. An indirect blocking ELAA (ib-ELAA) for M. gallisepticum antibodies detection was also developed using Apt-236, in which M. gallisepticum antibodies would block the binding-position of aptamers. Therefor positive sera would prevent color development whereas negative sera will allow a strong color reaction. The ib-ELAA was consistent with other three widely used assays in terms of the growth and decline of the antibody response to M. gallisepticum, and showed substantial agreement with the results obtained using a commercial ELISA kit in clinical chicken sera samples. Therefore, the ib-ELAA developed in this study was a new format for aptamer application and would be an alternative method for the surveillance of M. gallisepticum.

Introduction: Early childhood caries is a multifactorial disease. Saliva plays an important role in initiation and protection against caries, and its composition is greatly affected by nutritional status. This study was conducted to determine the impact of salivary lactoperoxidase and histatin-5 on the severity of ECC in relation to nutritional status. Materials and methods: The sample consisted of 120 children aged 5 years, classified into eight groups: mild ECC in underweight children, mild ECC in normalweight children, moderate ECC in underweight children, moderate in ECC normal weight children, severe ECC in underweight children, severe ECC in normalweight, caries-free (control) underweight children and caries-free normalweight children. Each group consisted of 15 children. Stimulated saliva was collected. Salivary lactoperoxidase was analysed using Human LPO/ Lactoperoxidase ELISA Kit (CLIA)-LS-F29892, and salivary histatin-5 was analysed using Human Histatin-5 ELISA Kit MBS705083_48T. Results: Lactoperoxidase and histatin-5 concentrations were significantly higher in caries-free children than in children with ECC, and they were higher in children with mild ECC than in children with moderate ECC or in children with severe ECC. They were significantly higher among children with normal weight than among those who were underweight (p 0.01), but there was significant interaction between these two variables and LPO and HST-5 together (p

The cross-sectional seroprevalence study of the peste des petits ruminants (PPR) in sheep and goats was carried out in the Southern Peninsular region of India to ascertain the prevalence of PPR virus (PPRV) antibodies at the epidemiological units (epi-units) level in the small ruminant population. The serum samples were collected from various epi-units (villages) in the different states and union territory (UT) in Southern Peninsular region using a stratified random sampling methodology from August 2017 to March 2018. A total of 6643 serum samples [sheep (n = 2785) and goats (n = 3858)] were collected from 360 epi-units and were screened by PPR competitive ELISA kit for the detection of PPRV antibodies. The results revealed that the seroprevalence of PPR in small ruminants in Telangana, Andhra Pradesh, Karnataka, Tamil Nadu, and Kerala states, and Puducherry UT was 87.0%, 66.4%, 64.3%, 47.8%, 11.4%, and 50.4%, respectively in the studied region. Further, the results of the chi-squared test revealed that the PPRV antibodies across different states and UT in the region were associated (sheep-χ(2) = 218.8, p

El objetivo fue evaluar la frecuencia serológica individual y de rebaño, así como detectar los factores de riesgo de C. abortus, en siete estados donde se realiza la producción ovina en México. Se efectuó un estudio multifactorial, transversal y estratificado, con análisis de 5,321 muestras serológicas en 323 rebaños de 61 municipios. Se realizó ELISA con un kit comercial. Los factores de riesgo asociados con la enfermedad se determinaron mediante cuestionarios y análisis estadísticos con una prueba de Ji cuadrada con un intervalo de confianza del 95%. Del total de 5, 231 muestras de suero que fueron colectadas, 581 (10.92 %) dieron positivas a la prueba de ELISA. Los resultados por estado de sueros positivos fueron: Tlaxcala 13.08% (73/558); Sonora 12.45 % (102/819); Chihuahua 11.56 % (107/925); Hidalgo 11.34 % (97/855); Chiapas 10.15 % (60/591); Querétaro 9.69 % (79/815); Estado de México 7.09 % (63/758). La frecuencia de hatos seropositivos, fue de 43.34 % (140/323). Los resultados al ser agrupados por estado fueron los siguientes: Hidalgo 67.39 % (31/46), Querétaro 67.18 % (43/64); Sonora 40.92 % (19/47); Tlaxcala 33.33 % (12/36); Chiapas 31.57 % (12/38); Estado de México 25.45 % (14/55) y Chihuahua 24.32 % (9/37). Los principales factores de riesgo que favorecen la presencia del aborto enzoótico de las ovejas, fueron que estuviesen gestantes, que tuviesen una edad entre los 37 a los 48 meses y que el sistema de producción fuese intensivo. Estos estudios serológicos identificaron la presencia del aborto enzoótico de las ovejas en México, así como algunos de los factores de riesgo que inciden para favorecer su presencia.

Background While there is evidence of a link between irisin and bone metabolism, prior clinical evaluations have been limited to women with postmenopausal osteoporosis. The primary goal of this evaluation is to examine the relationship between irisin and bone mineral density (BMD) in geriatric Chinese men experiencing osteoporosis or osteopenia. Methods In this case–control study, 43 geriatric Chinese men were verified as having osteoporosis or osteopenia via dual-energy X-ray light absorption spectrophotometry, and 24 subjects were accepted as the controls. Serum irisin levels were detected by a commercial ELISA kit. Results Serum irisin levels were lowered in geriatric Chinese men with osteopenia and osteoporosis, and multiple linear regression analysis revealed that the serum irisin level is an independent factor impacting BMD. Conclusions and discussion Our data confirm a positive correlation between irisin levels and BMD in geriatric Chinese men. Irisin has a protective effect on bone health dependent on BMD, but large clinical trials are still required to verify the irisin and BMD relationship.

Paratuberculosis (Ptb) is a widespread chronic infection caused by Mycobacterium avium subsp. paratuberculosis (Map) that affects both domestic and wild ruminants. Most of the studies focused on cattle while, the prevalence of Ptb in sheep in different regions of the world is not well investigated. This study aimed to address this gap of knowledge by screening adult female sheep for paratuberculosis antibodies in different geographical regions of Tunisia. A total number of 338 female sheep from 15 small to middle‐sized, extensively managed sheep farms in six regions across Tunisia were sampled. Animals were clinically examined before blood sampling. Sera were tested for the presence of anti‐Mycobacterium avium subsp. paratuberculosis antibodies using a commercial ELISA kit. Six farms out of 15 comprised at least one seropositive animal and 11 female sheep out of 338 tested animals (3.25%; 95% CI = [1.83–5.73]) were seropositive to M. avium subsp. paratuberculosis. The seroprevalence was significantly lower in 5‐year‐old females (p = .04) and animals that do not graze (p = .02). Due to its huge economic and social impacts, paratuberculosis represents a health problem in Tunisia and several other countries Further investigations are needed to rank sheep Ptb in Tunisia among other diseases and to assess the main risk factors using a larger nation‐wide survey., Six farms out of 15 comprised at least one seropositive animal and 11 female sheep out of 338 tested animals (3.25%; 95% CI = [1.83–5.73]) were seropositive to M. avium subsp. paratuberculosis, The seroprevalence was significantly lower in 5‐year old and older females (p = .04) and animals that do not graze (p = .02). This findings confirm that Paratuberculosis is circulating in sheep herds in Tunisia.

We examined anti-SARS-CoV-2 IgG and IgM antibodies in 45 serum samples from 26 patients with COVID-19, who were admitted in our hospital by using three different ELISA kits. All patients had pneumonia at admission, and 7 patients required mechanical ventilator support and grouped in severe case. Anti-SARS-CoV-2 IgG and IgM antibodies turned to be partially positive between the 6th and 10th days, more than 84% positive between the 11th and 15th days, and 100% after the 16th day. One ELISA kit revealed poorer sensitivity for anti-SARS-CoV-2 IgM antibody. Negative conversion of IgM antibody was not observed in the 30th day in our cohort. All three ELISA kits showed no false positive reaction for negative serum samples. Between severe and moderate cases, there was no significant difference in the trends of anti-SARS-CoV-2 IgG and IgM antibody.

A cross sectional study was conducted during the period 2014-2015, to estimate the prevalence and assess contribution of some risk factors for the occurrence of enterotoxaemia in sheep, goats, cattle and camels in the Kingdom Saudi Arabia. A total of 1593 animals from 476 herds were investigated. ELISA tests were conducted using a commercial kit for the detection of different toxins produced by Clostridium perfringens causing the disease. The toxicogenic typing of C. perfringens in clinical samples by ELISA kit, revealed that the predominant types were C. perfringens type A (67.2%) followed by type D (16.4%), then type B (13.4%) and type C (3%). Further confirmation of representative C. perfringens isolated from enterotoxaemia positive animals was performed using polymerase chain reaction. The overall prevalence of enterotoxaemia disease at the animal and herd level was 27.2% and 26.47% respectively. There were significant differences in the prevalence of enterotoxaemia in animals between the different regions in KSA, animal species and months of the year. The highest prevalence was detected in Aljouf region (41.7 %), followed by Hail region (40.9%), Qassim (37.8%), Jazan (31.1%), the eastern region (26.1%) and Riyadh region (10.4%). The prevalence was the highest cattle (64.3%), followed by goats (29.9%), camels (21.5%) and sheep (21.4%). Risk factors such as regions (χ2 = 89.65, p = 0.000), months of the year (χ2 = 76.65, p = 0.000), animal species (χ2 = 50.81, p = 0.000), disease presence by years (χ2 = 29.75, p = 0.000), herds prevalence (χ2 = 1443.6, p = 0.000) showed statistically significant association with enterotoxaemia disease. In the multivariate analysis, species of animals (OR = 4.88, p = 0.000) was found to be the most statistically significant risk factor associated with enterotoxaemia. Cattle were found to be approximately five times more likely to have infection with enterotoxaemia (OR = 4.88, CI = 2.55-9.37, p=0.0001) compared to other animal species. High overall prevalence of enterotoxaemia cases in all animal species occurred during the period August – December of the year 2015 with the highest prevalence in September (50%).

Objective To explore whether auricular concha eletroacupuncture (ACEA) is effective in regulating lipid-lipoprotein metabolism in rats submitted to cold stress. Methods Thirty-six adult male Sprague–Dawley rats were placed in four groups(9 rats in each group), the rats in three groups of which were submitted to cold stress for fourteen days, the last one of which was a control group. After the cold stress process, in those three groups, the rats of one group were with no treatment (stress only), two were treated with either ACEA or auricular margin eletroacupuncture (AMEA) repeated for fourteen days. On the 14th day, all the rats were sacrificed after all experimental procedure for blood sampling. Blood glucose, triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were tested by using the collected serum. Plasma lecithin-cholesterol acyltransferase (LCAT) was measured with ELISA kit. Results ACEA down-regulated the TG level (P Conclusion ACEA played an important role in regulating lipid-lipoprotein metabolism in rats submitted to cold stress.

AimTo explore the impact of puberty on refractive development and its interaction with outdoor time in children and adolescents.MethodsIn this 2-year observational study, students aged 7–13 years were selected with cluster sampling. All participants underwent cycloplegic refraction and axial length measurements once every year. Information of related factors was acquired through proper questionnaire or inquiry. The level of testosterone/estradiol was detected from the saliva of the subjects using the ELISA kit. Multiple linear regression and generalised estimating equation (GEE) were used to analyse the relationship among puberty, outdoor activities and refractive indicators.ResultsA total of 776 children and adolescents were included, with an average baseline age of 9.64±1.54 years and 53.6% boys. There were 350 myopes (55.2% of the 634 cyclopleged subjects) at baseline. There was a significant difference in the mean axial length changes and outdoor time among different puberty groups (for axial length: p=0.017, for outdoor time: p=0.015). Myopic parents, less outdoor time and more changes in estradiol were associated with greater changes in axial length and spherical equivalent (SE) (axial length changes: parental myopia β=0.230, outdoor time β=−0.250, changes in estradiol β=0.261; SE changes: parental myopia β=−0.267, outdoor time β=0.256, changes in estradiol β=−0.297). In the GEE model, the interaction between outdoor time and puberty was significantly associated with axial length (p=0.024, β=1.199).ConclusionsThis study implies puberty may play a regulating role on the relationship between outdoor time and refractive development among Chinese children and adolescents, which provides clues for in-depth mechanism interpretation and efficient intervention strategies.

Consumption of game in Germany has increased during the past 10 years. Wild boar (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) are the most frequently hunted and consumed game animals in Germany, yet information on the occurrence of zoonotic pathogens in these animal species is scarce. To better estimate the public health risk emanating from handling and consumption of game, this study investigated seroprevalences of Toxoplasma gondii in game hunted in the German federal state of Brandenburg during two hunting seasons from 2017 to 2019. Toxoplasma gondii-specific antibodies were detected in 24.4% (44/180, 95% CI: 18.4%-31.4%) of wild boar, 12.8% (16/125, 95% CI: 7.5%-20%) of roe deer and 6.4% (3/47, 95% CI: 1.3%-17.5%) of red deer using a commercial ELISA kit. Seroprevalences were similar in the two hunting seasons. Correlation between sex and seropositivity could not be observed. A rise in seroprevalence was seen with increasing age in all studied game species. Observed seroprevalences suggest that T. gondii is endemic in the sylvatic environment in the German federal state of Brandenburg and imply that game could represent a relevant source for human T. gondii infection.

Introduction: Cryptococcus neoformans is an encapsulated fungal pathogen that causes life-threatening meningoencephalitis in immunocompromised patients. This yeast secretes several potent immunogenic proteins by secretory vesicular mechanisms, such as HSP70 chaperone. Methods: The PCR-amplified HSP70 gene was cloned into a PET-28a(+) expression vector. The purified recombinant HSP70 (rHSP70) was evaluated by western blotting using an anti-His Tag-HRP antibody and then used for immunization of a rabbit. The serum of the immunized rabbit was tested against the whole lysate of C. neoformans in ELISA. Results: The antibodies in the rabbit’s serum recognized lysate of C. neoformans yeast. The highest antibody levels were achieved after the third booster injection. Conclusion: The rHSP70 showed to be a reliable candidate for the designing and development of an ELISA kit for early detection of cryptococcosis, and to screen a large number of specimens.

Sepsis is a cause of non-cardiac death in the hospital. Early and rapid diagnosis of septic patients is a challenge toincrease the expectancy of life. IL-6 and TNF-α are groups of pro inflammatory cytokines that initiate an initial inflammatoryresponse. Procalcitonin is a specific marker of bacterial infection. This study aimed to analyze the correlation of serumcytokine IL-6, TNF-α, procalcitonin and leukocyte count in suspected sepsis patients. This was a cross-sectionalobservational study consisting of 45 patients with suspected sepsis with procalcitonin level > 0.5 ng/mL. Procalcitonin levelwas measured with Enzyme-Linked Fluorescent Assay (ELFA) (VIDAS), IL-6 and TNF-α levels were measured with theU-CyTech Human Elisa kit (Bioscience, INC) and leukocyte counts were measured with SYSMEX-XN 1000. It was found in thisstudy that IL-6 levels ranged in 0 pg/mL – 73.29 ng/mL (mean 29.43 ng/mL), TNF-α levels were 0 pg/mL – 390.5 pg/mL (mean27.62 pg/mL), and the mean value of leukocytes was 20,139/μL. There was no correlation between leukocyte counts with IL-6(p=0.798 and r=0.040), TNF-α (p=0.304 and r = -0.160), and procalcitonin (p=0.323 and r = 0.154). There was no correlationbetween IL-6 levels with TNF-α levels (p=0.871 and r = -0.025), and procalcitonin levels (p = 0.466 and r = 0.112). There wasa weak negative correlation between TNF-α level and procalcitonin levels (p=0.006 and r = -0.403) and there was a weaknegative correlation between procalcitonin and TNF-α levels in suspected sepsis patients.

This study aims to investigate the concentration of traffic air pollution and the level of testosterone, fasting glucose, cortisol, and lipid profile among traffic police in Jakarta. Testosterone plasma and blood biochemicals in traffic police and the police officer were analyzed using ELISA kit. Air quality data from the Regional Environmental Agency (BPLHD) Jakarta 2012 were used for the characterization of exposure. The analysis used a t-test and linear regression test to show the relationship levels of pollutants exposure with the effects of reproductive dysfunction (p

Introduction: The U-Dream line of products are marketed as natural health product sleep aids in Canada and as dietary supplements in the United States. Several user reviews of the product mention concerning side effects not typically associated with the listed herbal ingredients stated on the product label. Based on these concerns an analytical study was undertaken to determine if the products contained any undeclared pharmaceuticals. Methods: Product was screened by high resolution mass spectrometry (HRMS) for known pharmaceuticals with sedative and sleep promoting properties. Based on the mass spectral screens an unknown compound was isolated for characterization by nuclear magnetic resonance (NMR) and presence confirmed by Enzymelinked Immunosorbent Assay (ELISA). Results: The mass spectral analyses indicated the presence of an undeclared analogue of the pharmaceutical drug zopiclone within the product lot tested. NMR characterization confirmed the compound to be a brominated analogue of zopiclone and a commercial zopiclone/eszopiclone ELISA kit tested positive. Discussion: The undeclared compound was found to be an analogue of zopiclone whereby the chlorine atom was substituted with bromine. Given the results of the ELISA assay and the structural similarity to zopiclone it is likely the compound exhibits biological activity. Of considerable concern is not only the potential of the unknown compound to exhibit pharmacological activity, but the lack of a safety profile by which the risk can be properly assessed. Conclusions: The natural health product regulations provide a framework for high quality, safe and efficacious products to access the market. It is the responsibility of the manufacturer to assure traceability and transparency in their supply chain and establish verifiable compliance with GMP. This study illustrates the importance of careful evaluation of analytical data in order to detect undeclared adulterants and highlights the need for an active monitoring and surveillance system for potentially high-risk products.

Objective This study aimed to examine whether honey and propolis Trigona can reduce the level of leptin in central obesity human. Methods It was a quasi-experimental study, population and sample were central obesity and normal weight as control. Sample size was 30 participants taken using purposive sampling. Results Intervention as long as 14 days make an increased level of leptin as many as 97.076 pg/ml in control group and decreasing respectively 171.803 pg/ml and 245.293 pg/ml in honey and propolis intervention group, both significant statistically. The dose of honey given was 105 mg/days divided into 3 packs, and each pack contains 35 mg. The dose of propolis was 60 mg/days and divided into 3 capsules, each of it contains 20 mg. Intervention as long as 14 days and leptin levels examined using human ELISA Kit three times, that is before intervention, 7 days after the intervention and 14 days after intervention. Conclusion Honey and propolis Trigona has the ability to decrease leptin level of central obesity participants. These honey bee products are potential to be a dietary supplement for central obesity patients.

Objective The objective of the research was to determine the giving of Holothuria scabra on the level of Docoxahexaenoic Acid (DHA) of breast milk with preterm chilbirth. Methods The research was a quasi experimental study with control group design. The samples were postpartum mothers with preterm childbirth consisted of 40 people divided into two groups. Postpartum mothers given Holothuria scabra capsule were intervention group and the postpartum mothers not given Holothuria scabra were control group. The sample of mother's milk of pretest of each group was taken in the seventh day and seven day later the sample was taken 3 cc. Intervention was done for 7 days per sample with 3 times dosage consisted of 2 capsules per day. The sample of breast milk was examined with Human DHA ELISA kit. The data were analyzed using Mann–Whitney test and Wilcoxon test. Results The results of the research indicated the difference of DHA level between intervention group and control group with a value of p = value 0.006. After the intervention, the average increase was 187.02 μg/mL for intervention group and 7.05 μg/mL for control group with a value of 1151.04 μg/mL.

Allergic responses are mainly caused by IgE, which is often located on the cell surface. The current diagnostic method detects both allergen-specific IgE and total IgE levels, but a number of allergic patients have a normal serum IgE level, which is a poor clinical correlate for allergy. Here, we developed a simple method to detect the level of cell-bound IgE by dissociating it from blood cells with lactic acid. Dissociated cell-bound IgE and plasma IgE levels were detected using the same ELISA kit at the same time. We established two clinical cohorts: an allergic patient group and a healthy participant group. In general, cell-bound IgE correlated well with plasma IgE; however, some patients exhibited high cell-bound IgE levels but low plasma IgE levels. We recommended 350 ng/mL peripheral blood total IgE (cell-bound IgE + plasma IgE) as the cut-off value for allergy diagnosis. Using this indicator, 90.32% of our allergic patients were correctly diagnosed. The peripheral blood total IgE level is a promising clinical diagnostic indicator in allergic patients and will provide more guidance for allergy diagnosis and therapeutic evaluation.

Background Defective cellular elements constitute an important challenge to achieve predictable periodontal regeneration. In an attempt to improve the cellularity of periodontal defects, gingival fibroblasts were implanted without their associated extracellular elements in periodontal defects to expose them to periodontal tissue mediators. In order to investigate the regenerative potential of gingival fibroblasts translocated into periodontal defects, the present study was designed to clinically and biochemically investigate the use of gingival fibroblasts (GF) and their associated mesenchymal stem cells (GMSC) in the treatment of intrabony periodontal defects. Methods A total of 20 subjects were randomly divided into two groups (n = 20). Group I: ten patients were included with ten intrabony periodontal defects that received β-calcium triphosphate (β-TCP) followed by collagen membrane defect coverage, while group II: (10 patients) ten periodontal defects received cultured gingival fibroblasts (GF) on the β-TCP scaffold and covered by a collagen membrane. The clinical evaluation was carried out at the beginning and at 6 months. Gingival crevicular fluid (GCF) samples were collected directly from the test sites for the quantitative measurement of PDGF-BB and BMP-2 using the ELISA kit at 1, 7, 14, and 21 days after surgery. Results Group II reported a significantly greater reduction in vertical pocket depth (VPD) and CAL gain compared with group I after 6 months. Radiographic bone gain was statistically higher in group II compared with group I. A significantly higher concentration of PDGF-BB was observed in group II on days 1, 3, and 7 compared with group I. Conclusions Translocation of gingival fibroblasts from gingival tissue to periodontal defects could be a promising option that increases cellular elements with regeneration potential. The concept of total isolation of gingival fibroblasts using occlusive membranes must be re-evaluated.

Furazolidone, a banned nitrofuran antibiotic, is still illegally used in aquaculture and livestock husbandry. A commercial Enzyme-linked Immunosorbent Assay (ELISA) from Europroxima, The Netherlands is described, and validated for the screening of 3-amino-2-oxazolidinone (AOZ), metabolite of furazolidone antibiotic in the muscle of shrimp. Research works was conducted at Veterinary Drug Residue Analysis Laboratory, Institute of Food and Radiation Biology, Atomic Energy Research Establishment, Savar, Dhaka-1349, between July 2018 and December 2019. Validation was done in accordance with the European Commission (EC) decision 2002/657/EC in terms of specificity, detection capability CCβ, selectivity, ruggedness, and stability. Homogenized sample was subjected to acid hydrolysis for releasing of tissue bound metabolite AOZ, and derivatized with o-nitrobenzaldehyde followed by ethyl acetate/n-hexane extraction and detection by ELISA reader. The detection capability CCβ of the method was lower or at 0.3 µg/kg accepting 0% false compliant result. Cut-off factor Fm and the positivity threshold value T was 0.716 and 0.982, respectively. Mean OD of blank sample B was1.225. The assay was valid because T was greater than Fm and smaller than B. The kit was stable even after ten months of storage at recommended temperature and showed good sensitivity. From the practical point of view, the kit studied has good reliability, and could be used in the routine application for the control of illegal use of the drug.

Level of enzyme peptidyl arginine deiminase type 4 (PAD-4) was determined in serum samples collected from 98 females with primary breast cancer within January, 2017 – April, 2018 divided into 5 groups according to immunohistochemistry data: group 1 – luminal A cancer, 2 – luminal B Her-2 negative, 3 – luminal B Her-2 positive, 4 – non-luminal Her-2 positive, 5 – triple negative cancer. Samples were collected prior to the onset of any anti-cancer treatment, and in 41 cases – 15-24 days after performing radical surgery. Control samples were obtained from 20 healthy females. Serum PAD-4 level was measured with Human PAD-4 ELISA Kit (Wuhan Fine Biotech Co., Ltd, China) on automatic analyzer ADALTIS Personal LAB (Adaltis S.r.l., Italy). Statistical analysis was performed by using software IBM SPSS Statistics 19. Mean serum PAD-4 level before treatment was 9.0 ng/ml that did not change after surgery. However, mean PAD-4 level for healthy females was 1.5 ng/ml (0.0-2.0) ng/ml that significantly differed from cancer group (Mann–Whitney U test, U = 38.500, p0.001). in contrast PAD-4 level in various cancer groups was: group 1 – 11.05 ng/ml, group 2 – 11.9 ng/ml, group 3 – 10.8 ng/ml, group 4 – 7.99 ng/ml, group 5 – 9.9 ng/ml. Thus, level of serum PAD-4 was higher in groups with more favorable luminal cancer, but further research is needed to make definitive conclusions.

BACKGROUND. Adiponectin, vaspin and leptin are only a few of these numerous adipocytokines. Little is known about the behavior of adipocytokines and how adipose tissue metabolism is affected in this Type 1 DM model. In this study we investigated the serum levels of adiponectin, leptin, vaspin in streptozotocin(STZ) induced diabetic rats. MATERIAL AND METHODS. Twelve Spraque Dawley albino rats were included in the study. The animals were divided into two groups. The first group was diabetic (D) (n: 6) and 60mg / kg STZ was administered intraperitoneally (i.p.) to these rats. The second group was the non-diabetic control (ND) group (n: 6). All the animals were euthanized by cervical dislocation. Quantification of vaspin, Adiponectin, leptin in serum was performed using the ELISA kit. RESULTS. Adiponectin, vaspin levels of diabetic group were found to be statistically lower than of control group (p

Background: In September 2017, the AOAC INTERNATIONAL Stakeholder Panel for Alternative Methods adopted Standard Method Performance Requirement (SMPR®) 2017.021, “Quantitation of Wheat, Rye, and Barley Gluten in Oats,” as guidance for the validation of methods for measuring gluten in oat products. The SMPR requires prospective methods to demonstrate adequate recovery (50–200%) based on the analysis of a set of reference samples. Objective: This document provides specific methods and data on the preparation of such validation materials and their analysis by an R5 ELISA kit to demonstrate the SMPR recovery estimation procedure. Methods: Seven reference samples were made by spiking wheat, rye, and barley into gluten-free oat flour at two levels, 10 and 20 mg/kg. The levels of gluten were determined by a wet chemical method based on the Codex Alimentarius definition of gluten. Results: The recoveries for wheat, rye, and barley were 122, 425, and 349%, respectively, for the R5 ELISA kit. The wet chemical method for estimating gluten in a sample of pure grain demonstrated repeatability relative SDs ranging from 1.40 to 2.75%. Conclusions: The reference materials are suitable to estimate ELISA kit responses to wheat, rye, and barley and calculate recoveries. Highlights: A series of oat flours spiked with wheat, rye, and barley flours were developed to be used as reference materials. A wet chemical method was established to estimate gluten contents based on the Codex definition. The reference materials are available for purchase to support further method development and validation.

Introduction Physical activity is one important factor in the nervous system of animals, it may affect the structure also the function of the brain. Regular aerobic exercises have a good effect on the whole body and thus improves the sense of well-being. Aim The aim of this experiment was to evaluate the effect of aerobic physical activity in white laboratory rats for 21 days. Methods The experimental animals used were twenty Wistar rats, divided into 2 groups of 10 animals they were subjected to forced swim test for 21 days for 60-90 min swim. The evaluation of levels of Adrenaline was performed on 1st, 7th, 14th day and on the last day of the experiment day 21 using ELISA kit protocol, also body mass was compared between groups. Results Our results showed that inducing aerobic physical activity for 21 days on the rats affects their levels of adrenaline. Comparatively, the control group of rats had significantly higher levels of adrenalin compared with day 21 (p=0.435) but lower compared with day 7 and 14 (p=0.231). There was also a difference in body mass which demonstrates adaptability to the surroundings and better coping with physical stress. Conclusion Regular aerobic activity for 21 days, for 60-90 min swim has a positive impact on adrenaline level also this aerobic exercise protocol could have a positive impact on reducing and maintaining body weight, thus preventing overweight.

Bacground. Obstructive sleep apnea-hypopnea syndrome (OSAHS) is associated with increased incidence and progression of atrial fibrillation (AF). Experimental studies indicate that OSAHS can modify the cardiac extracellular matrix (ECM). However, significant mechanisms of cardiac remodeling in AF patients with OSAHS have not been uncovered. The aim: to evaluate the impact of OSAHS on serum levels of tissue inhibitor of matrix metalloproteinase-1(TIMP-1) and hypoxia-inducible factor-1α (HIF-1α) in AF patients with ischemic heart disease (IHD) and/or arterial hypertension (AH), suffering from OSAHS. Material and methods. We examined 140 non-valvular AF patients with IHD and/or AH. Cardiorespiratory monitoring was conducted in the studied subjects and they were divided into 2 groups according to the presence of OSAHS: group 1 – non-OSAHS patients; group 2 – patients with OSAHS. Serum levels of TIMP-1 and HIF-1α were assayed using commercially available ELISA kit. Results. The patients with OSAHS had higher levels of TIMP-1 and HIF-1α than non-OSAHS patients (p