Enzyme Immunoassay Kit Research Articles

Plasma calcitonin gene-related peptide (CGRP) in migraine and endometriosis during the menstrual cycle.

ObjectiveMigraine, endometriosis, and the comorbidity of both are frequent pain disorders of special relevance for women. The neuropeptide calcitonin gene‐related peptide (CGRP) is critically involved in migraine, and circumstantial evidence suggests a role in endometriosis. We assessed CGRP levels at different times of menstrual cycle in four groups: healthy women, women with migraine or endometriosis and with the comorbidity of both.MethodsWomen with episodic migraine and women with a histologically confirmed endometriosis were recruited from specialized centers. For CGRP determination with a commercial enzyme immunoassay kit, cubital vein blood samples were collected on menstrual cycle day 2 ± 2 (during menstruation) and on day 15 ± 2 (periovulatory period). The primary endpoint of the study was the absolute difference of CGRP plasma levels between the menstrual and the periovulatory phase of all study groups. Groups were compared using nonparametric test procedures.ResultsA total of 124 women were included in the study. The change of CGRP plasma levels between menstruation and the periovulatory period was different between groups (p = 0.007). Women with comorbid migraine and endometriosis showed an increase of CGRP in the menstrual phase of +6.32 (interquartile range, IQR −3.64–13.60) compared to the periovulatory time, while healthy controls had a decrease of −10.14 (−22.54–0.91, p = 0.004). CGRP levels were different in the periovulatory phase among groups (p = 0.008), with highest values in healthy controls.InterpretationCGRP levels change significantly during the menstrual cycle. Different patterns in women with the comorbidity point to a deviant regulation of CGRP release.

Адіпонутрін як маркер метаболічних порушень у пацієнтів на гіпертонічну хворобу з надмірною масою тіла та ожирінням

Hypertension is a leading risk factor for coronary heart disease, stroke, peripheral arterial disease, kidney disease, vascular cognitive impairment, and mortality worldwide. Obesity in combination with hypertension continues to attract considerable attention from scientists. Despite the presence of experimental and clinical evidence of mutual burdening of hypertension and obesity, the mechanisms of formation of pathophysiological consequences of such a combination remain insufficiently defined. That is why in recent years an important role is given to the adiponutrin protein. The purpose of the study is to assess the nature of the relationship between adiponutrin levels and lipid and carbohydrate metabolism depending on the duration and degree of hypertension and the degree of obesity in patients with this comorbid pathology. Material and methods. The study included 107 patients with hypertension who were hospitalized in the cardiology department of Municipal noncommercial enterprise "Kharkiv City Clinical Hospital No.27" of Kharkiv municipal council. The age of patients ranged from 32 to 79 years (mean age was 58.6±9.88), who had not previously received regular antihypertensive therapy. The control group consisted of 20 age- and sex-appropriate healthy individuals. The level of adiponutrin was determined in 58 patients with hypertension of the 1st - 3rd degrees, aged from 32 to 79 years (mean age was 57.5±10.11 years), while hypertension of the 1st degree was diagnosed in 12 (20.7%) patients, hypertension of the 2nd degree - in 16 (27.6%), and hypertension of the 3rd degree - in 30 (51.7%) of the examined patients. Blood pressure was measured in patients in a sitting position after 5 minutes of rest, anthropometric measurements were performed, lipid profile and indicators of carbohydrate metabolism were evaluated, adiponutrin levels (pg/mL) were determined by enzyme immunoassay method using The RayBio® Adiponutrin Enzyme Immunoassay (EIA) Kit, (USA). Results and discussion. In all groups of patients with hypertension with overweight and obesity, adiponutrin levels were significantly elevated, and lipid and carbohydrate profiles tended to increase. Significant direct correlations were found between adiponutrine and triglycerides, with very low-density lipoprotein cholesterol of a moderate nature, and draws attention with a very high body mass index. In considering the differences in adiponutrin levels depending on gender, it was found that men have higher adiponutrin levels than women. Conclusion. Thus, we have the opportunity to consider adiponutrin as a marker of metabolic disorders

Plasma Osteopontin Correlates with Glycemic Control in Type 2 Diabetes Mellitus Patients

Diabetes Mellitus (DM) is a metabolic disease characterized by hyperglycemia due to abnormal secretions and/or insulin activity. Osteopontin (OPN) is an important component of inflammation and insulin resistance, and vitamin D decreases insulin resistance. This study aimed to analyze the correlation between OPN and glycemic control and total 25-OH vitamin D in type 2 DM. An observational analytic study with a cross-sectional approach was performed in Dr. Moewardi Hospital, Surakarta, from May to September 2018. Plasma OPN levels were measured by a sandwich enzyme immunoassay kit from Elabscience 96T Human OPN (USA), and a total of 25-OH vitamin D was evaluated using the ELFA method from Biomerieux SA (France). Data were tested by Pearson correlation (r). Type 2 DM subjects consisted of 45 (54.2%) males and 38 (45.8%) females, 36 (43.45%) well- and 47 (56.65%) poorly-controlled. The average age was 56.81±9.76 years old. The mean of OPN level in poorly-controlled cases was significantly higher (20.27±3.20 ng/mL) than well-controlled ones (15.04±3.34 ng/mL) with p=0.001. There was no significant difference in total 25-OH vitamin D between well- and poorly-controlled groups (19.84±6.65 vs. 17.24±6.78 ng/mL, respectively, p=0.085). The correlation of OPN with glycemic control (fasting glucose, 2-hour post-prandial glucose, HbA1c) and total 25-OH vitamin D in all subjects with type 2 DM were r=0.241 (p=0.028), r=0.378 (p=0.0001) r=0.529 (p=0.0001) and r=-0.151 (p=0.173), respectively. This study suggested that plasma OPN level was correlated with glycemic control but not with serum total 25-OH vitamin D in type 2 DM. Further research was needed in populations of other types of DM and other research variables related to inflammation or insulin resistance.

Development of in-house ELISAs for the detection of anti-SARS‑CoV‑2 RBD and N IgG and IgM antibodies in biological samples

By the end of year 2019, the new virus SARS-CoV-2 appeared, causing the Coronavirus Disease 2019 (COVID-19), and spread very fast globally. A continuing need for diagnostic tools is a must to contain its spread. Till now, the gold standard method, the reverse transcription polymerase chain reaction (RT-PCR), is the precise procedure to detect the virus. However, SARS-CoV-2 may escape RT-PCR detection for several reasons. The development of well-designed, specific and sensitive serological test like enzyme immunoassay (EIA) is needed. This EIA can stand alone or work side by side with RT-PCR.In this study, we developed several EIAs including plates that are coated with either specially designed SARS-CoV-2 nucleocapsid or surface recombinant proteins. Each protein type can separately detect anti-SARS-CoV-2 IgM or IgG antibodies. For each EIAs, the cut-off value, specificity and sensitivity were determined utilizing RT-PCR confirmed Covid-19 and pre-pandemic healthy and other viruses-infected sera. Also, the receiver operator characteristic (ROC) analysis was performed to define the specificities and sensitivities of the optimized assay. The in-house EIAs were validated by comparing against commercial EIA kits. All in-house EIAs showed high specificity (98–99%) and sensitivity (97.8–98.9%) for the detection of IgG/IgM against RBD and N proteins of SARS-CoV-2.From these results, the developed Anti-RBD and anti-N IgG and IgM antibodies EIAs can be used as a specific and sensitive tool to detect SARS-CoV-2 infection, calculate the burden of disease and case fatality rates.

Testosterone and lean mass show a positive correlation with the technical performance of footballers.

Monitoring technical performance during a soccer match has become an indispensable practice, since the players perform different functions within the tactical system adopted by the team, resulting in specific technical actions that are fully responsive to the individual technical level of each player. Thus, current study aimed to investigate the possible direct and mediating relationship between salivary testosterone and the technical performance of semi-professional footballers during a training game. To characterize the sample, the anthropometric profile, body composition, biological maturation, lower limb power, speed, estimation of maximum oxygen consumption, and physical performance were used. To determine testosterone concentrations, saliva was collected before and after the training game, for subsequent measurement using enzyme immunoassay (ELISA) and Salimetrics Kits. For quantification of technical performance and participation with the ball during the training game, scout software was used from the footage of the training game. The reliability of the technical performance analyses was determined through the repetition test of two attempts (Cohen's Kappa agreement index). The direct relationship between postgame testosterone and total participation with the ball in the second half; the direct relationship between the percentage of lean mass with postgame testosterone and an apparent relationship between percentage of lean mass and total participation with the ball from the set of relations mediated by postgame testosterone concentrations. The results suggest that testosterone concentrations combined with high levels of lean mass may have an important role in the technical performance and participation with the ball of semi-professional footballers.

Serum Levels of Fibroblast Growth Factor 21 Are Positively Associated with Aortic Stiffness in Patients with Type 2 Diabetes Mellitus.

Aortic stiffness (AS), assessed using carotid–femoral pulse wave velocity (cfPWV), is associated with cardiovascular disease in type 2 diabetes mellitus (T2DM). The relationship between serum fibroblast growth factor 21 (FGF-21) and AS in T2DM patients was evaluated. Fasting serum FGF-21 levels of 130 T2DM patients were measured using an enzyme immunoassay kit. A validated tonometry system was used to measure cfPWV (>10 m/s indicated AS). Of these T2DM patients, 34.6% were defined as the AS group. T2DM patients with AS were older; exhibited higher systolic blood pressure, diastolic blood pressure, and body fat mass; higher triglyceride, fasting glucose, glycosylated hemoglobin, and creatinine levels; higher urine albumin-to-creatinine ratios and serum FGF-21 levels; and lower estimated glomerular filtration rates. The FGF-21 level (odds ratio = 1.005, 95% confidence interval: 1.002–1.009, p = 0.002) as well as systolic blood pressure was an independent predictor of AS and positively correlated to cfPWV values (β = 0.369, p < 0.001) in T2DM patients. For T2DM patients, serum FGF-21 level could be a predictor for AS.

Serum GDF15 Level Is Independent of Sarcopenia in Older Asian Adults

Background: Growth differentiation factor 15 (GDF15), induced by tissue inflammation and mitochondrial stress, has received significant attention as a biomarker of mitochondrial dysfunction and has been implicated in various age-related diseases. However, the association between circulating GDF15 and sarcopenia-associated outcomes in older adults remains to be established. Aim: To validate previous experimental data and to investigate the possible role of GDF15 in aging and muscle physiology in humans, this study examined serum GDF15 levels in relation to sarcopenia-related parameters in a cohort of older Asian adults. Methods: Muscle mass and muscle function-related parameters, such as grip strength, gait speed, chair stands, and short physical performance battery score were evaluated by experienced nurses in 125 geriatric participants with or without sarcopenia. Sarcopenia was diagnosed using the Asian-specific cutoff points. Serum GDF15 levels were measured using an enzyme immunoassay kit. Results: Serum GDF15 levels were not significantly different according to sarcopenia status, muscle mass, muscle strength, and physical performance and were not associated with the skeletal muscle index, grip strength, gait speed, time to complete 5 chair stands, and short physical performance battery score, regardless of adjustments for sex, age, and BMI. Conclusions: These findings indicate that the definite role of GDF15 on muscle metabolism observed in animal models might not be evident in humans and that elevated GDF15 levels might not predict the risk for sarcopenia, at least in older Asian adults.

Characterization of antibodies in human immunoglobulin products from different regions worldwide

AimThe antibody levels against a broad spectrum of pathogens were assessed in commercial intravenous immunoglobulin (IVIG) manufactured from pooled plasma obtained from different global regions. MethodsTwenty-four IVIG commercial lots from eight manufacturers corresponding to 12 brands were analyzed. The plasma was collected in 10 countries/regions. Depending on each pathogen, antibody levels were measured using specific commercial IgG-specific enzyme immunoassay kits or by cell culture neutralization test and guinea pig skin neutralization test. A principal component analysis was performed. ResultsFor polio and diphtheria (reference markers of the US authorities), all IVIGs had relevant titers in accordance with reference levels. IVIGs from Canada, Australia, and the USA were positive for titers against globally distributed pathogens or those under vaccination programs in the developed world (parainfluenza, Epstein–Barr, varicella-zoster, influenza B, parvovirus B19, and measles viruses). IVIG from Taiwan and Hong Kong showed low antibody titers for these pathogens but high titers for Pseudomonas aeruginosa. IVIG from India had high titers for pathogens frequently found in developing countries (West Nile, dengue, chikungunya, and hepatitis E viruses and Streptococcus pneumoniae). IVIGs from Argentina, Spain, Israel, and Czechia showed intermediate antibody concentrations. ConclusionThe antibody profile in IVIG was greatly influenced by regional characteristics including climate, vaccination programs, and the prevalence of pathogens in the different countries and regions.

Soroepidemiological survey of chlamydia abortus infection in bovine in the State of Pernambuco, Brazil

The objective of this study was to conduct a seroepidemiological survey of Chlamydia abortus infection in dairy cattle herds. A total of 303 blood serum samples were collected from 24 property in Vale do Ipanema microregion in the state of Pernambuco, Brazil. For the diagnosis of C. abortus infection, a commercial enzyme immunoassay kit (ELISA) was used. A prevalence of 34.0% (103/303; 95% CI: 28.7%-39.7%) of infected animals was identified. In 79.8% (19/24) of the properties, at least one infected animal was detected. The risk factors identified were: semi-intensive system (OR = 3.47, p ≤ 0.000), extensive system (OR = 8.14; p ≤ 0.000), supply of water in troughs and directly at the fountain (OR = 2.29, p = 0.002), pasture rent (OR = 1.72, p = 0.041), use of artificial insemination (AI) (OR = 3.07, p = 0.002), and use of AI associated with natural mount (OR = 2.22, p = 0.003). The occurrence of C. abortus infection in dairy cattle in the state of Pernambuco, Brazil, was recorded for the first time. It is concluded that the infection by this agent is present in the analyzed herds and that hygienic and sanitary management measures based on the identified risk factors should be implemented to avoid reproductive losses and losses to the producers.

HEV prevalence and potential risk factors in a large multi-ethnic youth cohort in China

BackgroundThis cohort study was designed to investigate the prevalence of and potential risk factors of HEV infection in a large multi-ethnic youth cohort in China.MethodsBlood samples were collected from participants (n = 6269) and serum was isolated. All serum samples were tested for anti-HEV IgG, anti-HEV IgM antibodies using commercial enzyme immunoassay kits (Wantai Biological Pharmacy Enterprise, Beijing, China).ResultsThe overall rate of anti-HEV IgG and anti-HEV IgM prevalence was 4.78% and 0.14%, 0.03% were positive for both anti-HEV IgG and anti-HEV IgM antibodies. Anti-HEV IgG positivity is significantly higher in females (5.27%) compared to males (4.14%) (P = 0.028). Anti-HEV IgG prevalence is significantly (P = 0.0001) higher in Dong (17.57%), Miao (12.23%), Yi (11.04%), Gelao (9.76%), and Bai (10.00%) compared to other ethnic groups. It is significantly higher in Guizhou (11.4%), Sichuan (10.1%), Yunnan (9.3%), and Guangxi (6.9%) than that other province. We found that ethnicity and provincial background are significantly associated with HEV infection in this cohort.ConclusionThis study provides comprehensive information on HEV prevalence in multi-ethnic populations in China. However, our study only focused on a youth population from different provinces of China. Future studies are recommended to investigate HEV prevalence in other age groups of the ethnic populations.

Targeted Temperature Management Suppresses Hypoxia-Inducible Factor-1α and Vascular Endothelial Growth Factor Expression in a Pig Model of Cardiac Arrest.

BackgroundThe hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF)/VEGF receptor subtype 2 (VEGFR-2) pathway has been implicated in ischemia/reperfusion injury. The aim of this study was to clarify whether whole-body hypothermic targeted temperature management (HTTM) inhibits the HIF-1α/VEGF/VEGFR-2 pathway in a swine model of cardiac arrest (CA) and cardiopulmonary resuscitation (CPR).MethodsTwenty-four domestic male Beijing Landrace pigs were used in this study. CA was electrically induced with ventricular fibrillation and left untreated for 8 min. Return of spontaneous circulation (ROSC) was achieved in 16 pigs, which were randomly assigned either to normothermia at 38 °C or to HTTM at 33 °C (each group: n = 8). HTTM was intravascularly induced immediately after ROSC. The core temperature was reduced to 33 °C and maintained for 12 h after ROSC. The serum levels of HIF-1α, VEGF, VEGFR-2, and neuron-specific enolase (NSE) were measured with enzyme immunoassay kits 0.5, 6, 12, and 24 h after ROSC. The expression of HIF-1α, VEGF, and VEGFR-2 in cerebral cortical tissue was measured by RT-PCR and Western blot analysis 24 h after ROSC. Neurological deficit scores and brain cortical tissue water content were evaluated 24 h after ROSC.ResultsThe serum levels of HIF-1α, VEGF, and VEGFR-2 were significantly increased under normothermia within 24 h after ROSC. However, these increases were significantly reduced by HTTM. HTTM also decreased cerebral cortical HIF-1α, VEGF, and VEGFR-2 mRNA and protein expression 24 h after ROSC (all p < 0.05). HTTM pigs had better neurological outcomes and less brain edema than normothermic pigs.ConclusionThe HIF-1α/VEGF/VEGFR-2 system is activated following CA and CPR. HTTM protects against cerebral injury after ROSC, which may be part of the mechanism by which it inhibits the expression of components of the HIF-1α/VEGF/VEGFR-2 signaling pathway.

Correcting for enzyme immunoassay changes in long term monitoring studies

Enzyme immunoassays (EIAs) are a common tool for measuring steroid hormones in wildlife due to their low cost, commercial availability, and rapid results. Testing technologies improve continuously, sometimes requiring changes in protocols or crucial assay components. Antibody replacement between EIA kits can cause differences in EIA sensitivity, which can hinder monitoring hormone concentration over time. The antibody in a common cortisol EIA kit used for long-term monitoring of stress in wildlife was replaced in 2014, causing differences in cross reactivity and standard curve concentrations. Therefore, the objective of this study was to develop a method to standardize results following changes in EIA sensitivity. We validated this method using cortisol concentrations measured in the hair of brown bears (Ursus arctos).•We used a simple linear regression to model the relationship between cortisol concentrations using kit 1 and kit 2.•We found a linear relationship between the two kits (R2 = 0.85) and used the regression equation (kit2 = (0.98 × kit1) + 1.65) to predict cortisol concentrations in re-measured samples.•Mean predicted percent error was 16% and 72% of samples had a predicted percent error <20%, suggesting that this method is well-suited for correcting changes in EIA sensitivity.

CSF Neurofilament-H Levels as a Potential Prognostic Marker in Patients of Guillain- Barré Syndrome- A Cohort Study

Introduction: The prognosis of Guillain-Barré Syndrome (GBS) at an early stage with explicit biomarkers is critical to distinguish patients with possibility of poor recovery. Cerebrospinal Fluid (CSF) serves as an impending source for biomarkers that portrays the exact biochemical changes. Aim: To find out if there is any prognostic value of high CSF phosphorylated Neurofilament Heavy subunit (pNf-H) levels, measured during first two weeks of onset of GBS, as assessed by the level of disability at six months after the onset of GBS. Materials and Methods: The cohort study was conducted in the Department of Neurology and Department of Immunology and Molecular Medicine, at the Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India, over a period of two years from August 2015 to August 2017. Sixty two patients who satisfied the required diagnostic standards for GBS (study group) and 35 patients with tension-type headache (control group) were selected for the study. After clinical and electrophysiological assessment, CSF samples were collected. A commercially available sandwich enzyme immunoassay kit, manufactured by BioVendor-Laboratorní medicína (Czech Republic), was used for measuring human pNf-H quantitatively. Results: Mean CSF pNf-H level in patients with good outcome was 325.3 pg/mL whereas, in patients with poor outcome it was 3655.2 pg/mL. CSF pNf-H levels were found to be suggestively higher in GBS patients with poor outcome as compared to those with good outcome. Only eight patients in good outcome group had pathologically high CSF Nf-H levels whereas 10 patients in poor outcome group had CSF Nf-H levels ≤730 pg/mL. The odds ratio was 17.1 (95% Confidence Interval (CI) 3.83-76.29). Thus, high CSF Nf-H levels on admission predicted poor outcome in GBS (p-value &lt;0.001). Moderate degree of positive correlation was found between CSF Nf-H levels and outcome (F score) at six months (R=0.684; p-value &lt;0.001). Conclusion: It can be determined that higher values of CSF pNf-H in GBS (acute stage), could serve as a predictive marker indicative of poor prognosis.

Detection of steroid and thyroid hormones in mammalian teeth.

Endocrine tools can provide an avenue to better understand mammalian life histories and predict how individuals and populations may respond to environmental stressors; however, few options exist for studying long-term endocrine patterns in individual marine mammals. Here, we (i) determined whether hormones could be measured in teeth from four marine mammal species: narwhal (Monodon monoceros), beluga (Delphinapterus leucas), killer whale (Orcinus orca) and Atlantic walrus (Odobenus rosmarus rosmarus); (ii) validated commercially available enzyme immunoassay kits for use with tooth extracts; and (iii) conducted biological validations for each species to determine whether reproductive hormone concentrations in teeth correlated with age of sexual maturity. Tooth extracts from all species had measurable concentrations of progesterone, testosterone, 17β-estradiol, corticosterone, aldosterone and triiodothyronine (T3); however, cortisol was undetectable. Parallelism between the binding curves of assay kit standards and serially diluted pools of tooth extract for each species was observed for all measurable hormones. Slopes of accuracy tests ranged from 0.750 to 1.116, with r2 values ranging from 0.977 to 1.000, indicating acceptable accuracy. Biological validations were inconsistent with predictions for each species, with the exception of female killer whales (n = 2), which assumed higher progesterone and testosterone concentrations in mature individuals than immature individuals. Instead, we observed a decline in progesterone and testosterone concentrations from infancy through adulthood in narwhal (n = 1) and walruses (n = 2) and higher reproductive hormone concentrations in immature individuals than mature individuals in belugas (n = 8 and 10, respectively) and male killer whales (n = 1 and 2, respectively). While unexpected, this pattern has been observed in other taxa; however, further analytical and biological validations are necessary before this technique can be used to assess individual mammalian endocrine patterns.

Immunogenicity and Safety of Inactivated SARS-CoV-2 Vaccine in People Living with HIV: A Cross-Sectional Study

Background: To analyze the efficacy and safety of SARS-CoV-2 inactivated vaccine in people living with HIV (PLWH).Methods: A total of 143 PLWH were included in the study. All patients were confirmed with HIV-1 infection. We also enrolled 50 healthy individuals vaccinated with two doses of SARS-CoV-2 vaccine as controls. A commercially available magnetic chemiluminescence enzyme immunoassay kit was used to detected serum IgG and IgM against SARS-CoV-2.Findings: Serum levels of SARS-CoV-2-specific IgG were significantly higher in the control group than in the PLWH group (P=0.001). Overall, 76% of individuals in the control group achieved IgG seroconversion after vaccination compared with 58% in the PLWH group (P=0.024). The time after vaccination in IgG seronegative PLWH was significantly longer compared with PLWH with IgG seropositive (43.38 ± 34.96 vs 30.27 ± 20.12 days, P=0.005). In PLWH with IgG seropositivity, CD4+ T cell counts before antiretroviral therapy (ART) (P=0.015) and at IgG detection (PInterpretation: The inactivated COVID-19 vaccine appears to be safe with good immunogenicity in PLWH.Funding: This study was supported by Clinical Research Startup Program of Southern Medical University by High-level University Construction Funding of Guangdong Provincial Department of Education (No. LC2016PY003).Declaration of Interest: None of the authors have competing interests to disclose.Ethical Approval: The study was performed in accordance with the Declaration of Helsinki and was approved by the Institutional Ethics Committee of Nanfang Hospital (NFEC-2021-178).

Whiskers as a novel tissue for tracking reproductive and stress-related hormones in North Pacific otariid pinnipeds.

Keratinized tissues, including whiskers, are ideal for acquiring a record of physiological parameters. Most tissues provide a snapshot of physiological status; however, whiskers may support longitudinal sampling for reproductive and stress-related hormones, if hormones are incorporated as whiskers grow and concentrations change with physiological state. Whiskers from female Steller sea lions (Eumetopias jubatus) and northern fur seals (Callorhinus ursinus) were serially sectioned and pulverized and steroid hormones were extracted. Standard methods were used to validate enzyme immunoassay kits for cortisol, progesterone, 17β-estradiol and testosterone. All hormones were measurable in whisker segments from both species with progesterone concentrations showing cyclical patterns, which appear to signify previous pregnancies or luteal phases. Yearly progesterone concentrations were greater in years a pup was produced compared with years when no pup was observed. Free-ranging female Steller sea lions had reproductive rates between 0 and 1.0 (0.53 ± 0.33, n = 12) using a yearly progesterone concentration of 30pg/mg or greater to classify a reproductive year as producing a pup and below 30pg/mg as non-reproductive. Cortisol concentrations were greater near the root and rapidly declined, lacking any obvious patterns, throughout the rest of the whisker. Progesterone and testosterone concentrations were able to help determine sex of unknown individuals. Immunohistochemistry revealed that steroid hormones most likely do not leach out of whiskers based on the deposition patterns of progesterone and cortisol being present throughout the whisker length. Overall, measuring steroid hormones in whiskers can reveal individual reproductive histories over multiple years in sea lions and fur seals. Cyclical patterns of δ15N were useful for identifying periods of up to ~10years of growth within whiskers, and measuring both stable isotopes and hormones may be useful for differentiating periods of active gestation from diapause and potentially track multi-year reproductive histories of female otariids.

Levels of carboxylated and undercarboxylated osteocalcin in patients with type 2 diabetes

Introduction: Osteocalcin (OC) is a bone-derived protein that undergoes vitamin K-dependent carboxylation. The undercarboxylated form of the protein (ucOC) is released in the circulation during the process of bone resorption. Experimental studies on mice and rats have revealed that ucOC is involved in the regulation of energy homeostasis, linking in this way the bone, pancreas, and adipose tissue metabolism. Experimental studies suggest no hormonal role for the carboxylated form (cOC) of the protein. Aim: In the current study we aimed to examine the levels of OC in its carboxylated and undercarboxylated form in patients with type 2 diabetes and control subjects, and to compare the vitamin K status between the two groups. Materials and Methods: The present cross-sectional study involved a sample of 46 adults type 2 diabetes patients and a control group of 19 individuals. The carboxylated and undercarboxylated forms of OC were measured in serum by using highly sensitive sandwich-type enzyme immunoassay kits. Vitamin K status was evaluated by the ratio ucOC/cOC. Student’s two-tailed unpaired t-test was used to compare the groups. Results: UcOC and cOC serum levels were significantly lower in patients with type 2 diabetes compared to controls. We found no difference in the vitamin K status between the groups. Conclusion: Our results show that OC might be involved in the regulation of carbohydrate metabolism. In humans, it appears that the carboxylation state might not be essential for the hormonal role of the protein as in mice and rats.

Diagnostic Accuracy of Synovial Neopterin, TNF-α and Presepsin in Periprosthetic Joint Infection: A Prospective Study.

Due to the lack of specificity of conventional diagnostic tools, the prediction of periprosthetic joint infections (PJI) remains challenging. The purpose of this study was to evaluate the accuracy of synovial fluid neopterin, presepsin, and TNF-α as diagnostic parameters and to compare it to the biomarkers recommended in the 2018 definition of periprosthetic hip and knee infection. Between August 2018 and July 2019, a prospective cohort study was conducted in 80 patients with painful hip, shoulder, and knee arthroplasty. In addition to medical history, clinical and laboratory data were gathered. PJI was diagnosed based on the 2018 definition of periprosthetic hip and knee infection. Synovial joint fluid was analyzed for biomarker measurement using standard quantitative enzyme immunoassay kits. Fifty-three patients (66%) were classified as the aseptic group and twenty-seven patients (34%) as the PJI group. The mean levels of synovial fluid neopterin were significantly higher (p < 0.01) in the PJI group than those in the aseptic group (aseptic 8.3 ± 6.9 vs. PJI 20.9 ± 21.4 nmol/L). The average values of synovial fluid TNF-α and presepsin were not significantly higher in the PJI group than those in the aseptic group (presepsin: aseptic 0.13 ± 0.19 vs. PJI 0.11 ± 0.32 ng/mL, p = 0.08; TNF-α: aseptic 6.6 ± 7.3 vs. PJI 46.3 ± 123.2 pg/mL, p = 0.17). Synovial fluid neopterin was 59% specific and 74% sensitive with a cut-off value of 7.2 nmol/L. The sensitivity and specificity of synovial fluid TNF-α were 63 and 51% with a cut-off value of 3.9 pg/mL. Synovial fluid presepsin was 51% specific and 29% sensitive with a cut-off value above 0.06 ng/mL. Synovial fluid neopterin appears to a reliable diagnostic marker for detection of PJI. In contrast, synovial fluid TNF-α and presepsin are not suitable to exclude or diagnose PJI.

Rotavirus cases among Rotarix vaccinated children below five years during the 2018 outbreak in the Kingdom of Eswatini

Background: A large outbreak of diarrhoeal disease in children under 5 years was reported in four surveillance sentinel sites Mbabane Government Hospital, Raleigh Fitkin Memorial Hospital, Hlathikhulu Government Hospital and Good Shepherd Hospital. Rapid increase of number diarrhoeal cases, were reported from week 34 to 41 (August to October 2018). Approximately 1,535 children presented with acute diarrhea, vomiting and dehydration. Methods and materials: Diarrhoeal stool samples were routinely collected for surveillance from children less than 5 years of age to determine the etiology of the diarrhoea. A total of 139 stool samples were tested for rotavirus using enzyme immunoassay kit, Molecular characterization using polyacrylamide gel electrophoresis, semi nested reverse-transcription polymerase chain reaction, sequencing and phylogenetic analysis. Rotavirus negative stool samples were further tested for bacteria and parasites using molecular diagnostic assays. Results: The rotavirus positive samples were positive for group A rotavirus with a prevalence of (117/211) 53% with the age groups mostly affected being;<12 months = (68/117)58.1%, 3-24 months = 23.9% (28/117) and >24 months = 16.2%(19/117). The PAGE patterns observed were long RNA migration pattern. The G3P[8] 59/75(78.6%) rotavirus strain were mainly associated with the outbreak. Phylogenetic analysis of the partial length VP4 P[8] associated G3 clustered together with other P[8] strains detected in Africa and distantly related to vaccine strains (Rotarix™ and RotaTeq™). Pathogens that were observed in Eswatini were Shigella/Enteroinvasive E. coli 18% (11/61) was frequently detected followed by Campylobacter jejuni/coli 13% (8/61) and Giardia lamblia 13% (8/61). Conclusion: Majority of the children affected by the rotavirus received vaccination and the rotavirus genotype G3P[8] was the major cause of these outbreak. It also important to note that the G3P[8] that was circulating was the wild type stains not related to the vaccine strains. As evident from the Maximum likelihood phylogenetic tree generated, G3P[8] study strains clustered in different lineages as compared to the vaccine strains (Rotarix™ and RotaTeq™). Further studies on whole genome sequence analysis are required to determine the evolutionary relationship among the strains and surveillance of rotavirus among the infants should be continued while increasing the rotavirus vaccine coverange and add a booster dose to increase protection.

In vivo effects of nitrosyl hydrogen on cardiac function and sarcoplasmic reticulum calcium pump (SERCA2a) in rats with heart failure after myocardial infarction.

Abnormal Ca2+ circulation in cardiomyocytes is an important cause of decreased myocardial contractility in failing hearts. Nitroxyl hydrogen (HNO) can oxidize Ca2+ cycle-related proteins, alter their biological functions, promote Ca2+ recovery as well as release, and enhance myocardial contractility. In this study, we aim to observe the effect of nitrosyl hydrogen (HNO) on the cardiac function of rats with heart failure and elucidate the underlying mechanism. Twenty six male Wistar rats were randomly divided into heart failure group (HF group), Angeli's salt treatment group (HF + AS group) and sham operation group (Sham group). The HF + AS group rats were treated with HNO donor Angeli's salt by intraperitoneal injection of 1 mg/kg/d, and the rats in the HF group and the Sham group were given the same amount of normal saline. Cardiac function was measured by echocardiography before and after treatment. NT-proBNP was measured by enzyme immunoassay (ELISA) kit after treatment. Western blot were used to measure the expression of sarcoplasmic reticulum Ca2+-ATPase (SERCA) in protein levels in rats. The activities of SERCA2a were detected by the biochemical kit finally. We found that Nitrosyl hydrogen could significantly increase LVEF, +dp/dt, -dp/dt (P<0.05), significantly decrease NT-ProBNP and LVEDP (P<0.01), and significantly enhance the activities of SERCA2a protein (P<0.05). These findings suggest that Nitrosyl hydrogen could improve the cardiac function possibly by increasing protein activities of SERCA2a in rats.