Search

Background:Borrelia burgdorferi sensu stricto is the causative agent of Lyme disease (LD). Possible early symptoms include flu-like symptoms and erythema migrans and later, the risk of disruption of the nervous system, joints, and heart. A two-tiered testing method is employed for serological diagnostics. The Public Health Agency of Canada guidelines recommend that samples tested negative on first-tiered test need not be confirmed by second-tiered test. Due to the challenging nature of diagnosis leading to misconceptions among physicians about false negatives, confirmatory testing is requested despite the initial negative result. Methods: Hundred screen-negative Lyme patient samples from 2007 to 2016 were tested by Western blot (WB) second-tiered confirmatory test upon physician's request in British Columbia to study the first-tiered screening test sufficiency. Results: Those negative for first-tiered enzyme-linked immunosorbent assay were also negative by WB. Conclusion: Results demonstrate that confirmatory testing is not necessary on screen-negative samples. Hence, first-tiered test is sufficient to rule out LD. [ABSTRACT FROM AUTHOR]

The measurement of biomarkers in blood and excreta can enable immune status assessment and provide prognostic information on individual health outcomes. In this respect, the fecal measurement of secretory immunoglobulin A (sIgA), the primary mammalian antibody for mucosal defense, has recently received increased interest in a few anthropoid primates, but a fecal sIgA assay for use in strepsirrhine primates has not yet been reported. Here, we develop and analytically validate a cost‐effective in‐house sandwich enzyme immunoassay for the extraction and measurement of sIgA in feces of redfronted lemurs (Eulemur rufifrons). We also tested a simple method for sIgA extraction that can be used under remote field conditions and undertook experiments to assess the robustness of sIgA concentrations to variation in processing and storage conditions of fecal extracts. Our analytical validation revealed that the assay recognizes immunoreactive sIgA in redfronted lemur feces, that sIgA can be measured accurately with no potential interference from the fecal matrix, and that assay reagents and performance are highly stable over time. The field‐friendly extraction procedure produced sIgA results strongly correlated with those generated by a standard laboratory extraction method. Short‐term storage at room temperature resulted in a slight decline in sIgA concentrations, whereas freezing extracts at −20°C kept sIgA levels stable for at least 3 months. Longer‐term storage of >5 months, however, led to a significant decline of sIgA concentrations. Multiple freeze‐thaw cycles did not affect sIgA levels. This study, therefore, provides the basis for measuring fecal sIgA in lemurs and possibly other strepsirrhines. When samples are processed properly and stored frozen, and when sIgA analysis can be performed within 3 months upon sample collection, fecal sIgA measurements can become a valuable tool for monitoring aspects of immunity and health in both zoo‐housed and wild‐living lemurs. Research Highlights: We established and analytically validated a cost‐effective, in‐house ELISA for measuring fecal secretory immunoglobulin A (sIgA) in redfronted lemurs.This method ensures accurate results even under challenging field conditions, with stable sIgA concentrations up to 3 months under frozen conditions.SIgA as a non‐invasive biomarker provides a valuable tool for monitoring aspects of immunity and health in zoo‐housed and wild lemurs. [ABSTRACT FROM AUTHOR]

Antibodies to azo dyes of the Congo Red (CR) type, which bind to amyloid fibrils both in vitro and in vivo, were obtained in order to develop an approach to therapy of amyloidoses based on the use of antibodies to amyloid components. Since CR and its analogues do not have readily activated groups necessary for covalent binding of these dyes to protein antigens, azo dye containing a fragment of a symmetric CR molecule and a free carboxyl group was synthesized. This compound, obtained from para-aminobenzoic acid and naphthionic acid, is 4-(1-amino-4-sulfonaphthyl-2-azo)benzoic acid (ASABA). It was shown that ASABA can be activated by a mixture of N-hydroxysuccinimide and 1,3-dicyclohexylcarbodiimide and then covalently attached to proteins. Conjugates of ASABA with king crab hemocyanin (HC–ASABA) and bovine serum albumin (BSA–ASABA) were synthesized. HC–ASABA was used as an antigen for rabbit immunization. Polyclonal antibodies from rabbit blood sera recognized ASABA in the conjugates, as well as free CR and its fluorene-based analogue, 2,7-bis(1-amino-4-sulfonaphthyl-2-azo)fluorene. These antibodies can be used for further work on the immune destruction of amyloid stained with CR type dyes. [ABSTRACT FROM AUTHOR]

Background: The etiology of temporomandibular disorders (TMD) is multifactorial, involving a complex interplay of psychological and physiological factors. While salivary biomarkers, particularly cortisol, play an important role in TMD pathophysiology, evidence in the literature is still scarce and inconsistent. Hence, this study aims to evaluate the applicability of salivary cortisol as a potential biomarker for TMD in adults. Methods: The Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) were used to accurately diagnose TMD patients. The study included adults, both male and female, aged 18–40 in the TMD group (n = 66) and non TMD participants (n = 66) matched for age and gender. Salivary samples were collected from participants at two time points: early and late morning. Cortisol levels in the samples were quantified using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using a one-way ANOVA to evaluate the correlations between cortisol levels and the study variables. Tukey's post-hoc tests were applied to adjust for multiple comparisons. Results: Salivary cortisol levels were significantly higher in the TMD group than in the corresponding controls (p = 0.034). In the TMD group, the mean cortisol levels early in the morning were 29.95 ± 75.05 (0-398.64), while the late morning levels were recorded as 4.87 ± 3.96 (0-17.13). In the control group, the mean cortisol levels early in the morning were 10.98 ± 16.83 (2.16–92.90), and late morning mean cortisol levels were 6.15 ± 6.13 (0-20.42). This indicates that the early morning levels of cortisol are higher in TMD patients (p = 0.046). In the subgroup analysis of the TMD, the mean salivary cortisol levels recorded were highest at 82.49 ± 124.34 (8.32-398.64) in patients having disc displacement without reduction with limited mouth opening. Furthermore, the mean salivary cortisol levels in the early morning were statistically higher (84.83 ± 132.80) in males compared to females (9.36 ± 9.01) (p = 0.008) with TMD. Conclusion: The result of this study suggests that salivary cortisol could be a potential biomarker for a specific TMD subtype (disc displacement without reduction with limited mouth opening). However, further studies are needed to better understand the role of cortisol biomarker in the underlying pathogenesis of TMD. [ABSTRACT FROM AUTHOR]

Objective In evaluation of systemic lupus erythematosus (SLE), anti–double-stranded DNA antibodies (anti-dsDNA) play a significant role in diagnosis, monitoring SLE activity, and assessing prognosis. However, evaluations of the performance and limitations for recently developed methods for anti-dsDNA assessment are sparse. Methods Specimens used for antinuclear antibody testing (n = 129) were evaluated for anti-dsDNA assay comparability across 4 medical centers in the United States. The methods compared were Werfen Quanta Lite dsDNA, Zeus Scientific dsDNA Enzyme Immunoassay, Bio-Rad multiplex immunoassay (MIA) dsDNA, ImmunoConcepts Crithidia, and Bio-Rad Laboratories Crithidia. Results For quantitative anti-dsDNA measurements, Spearman's correlation coefficient was highest between Zeus and Werfen (ρ = 0.86; CI, 0.81-0.90; P <.0001). Comparison of MIA to Werfen or Zeus yielded similar results to each other (ρ = 0.58; CI, 0.44-0.68; P <.0001; and ρ = 0.59; CI, 0.46-0.69; P <.0001, respectively), but lower than the correlation between Zeus and Werfen. Positive concordance between assays ranged from 31.4% to 97.1%, and negative concordance between assays ranged from 58.5% to 100%. The detection of anti-dsDNA in those with SLE diagnosis ranged from 50.9% to 77.4% for quantitative assays and 15.1% to 24.5% for Crithidia assays. Conclusion Current quantitative anti-dsDNA assays are not interchangeable for patient follow-up. Crithidia -based assays demonstrate high negative concordance and lack positive concordance among the methods. [ABSTRACT FROM AUTHOR]

The purpose of the study was to investigate the cultural method of the diagnosis of Chlamydophila abortus. For this purpose, laboratory diagnosis of pathological material from the farm was carried out at the LLP Research and Production Enterprise "Antigen", where, based on the data of anamnesis, data of occurrence of similar epizootic picture for the last two years, examination and diagnostic manipulations (clinical symptoms, autopsy data, epizootological anamnesis), the preliminary diagnosis "Enzootic abortion of ewes caused by Chlamydophila abortus" was made. Samples from the organs of aborted foetuses and swabs with vaginal contents were taken to confirm this diagnosis. Complement fixation tests with chlamydia antigen were found to be positive in 7 (23%) of the 30 animals that were examined. To isolate the pathogen, the technology of suspension cultivation in a bioreactor of the Chlamydophila abortus strain on McCoy cell culture was used. Isolated pathological material was grown in McCoy cell cultures that were suspended in a bioreactor in Dulbecco's Modified Eagle's Medium (DMEM) nutrient medium with 10% foetal bovine serum and antibiotics at a concentration of 2×105 cells/mL. Enzyme immunoassay was a confirmation of the successful result of cultivation. Enzyme immunoassay confirmed the presence of the pathogen in 60.9% of cases. 25 samples were received, which were positive. Thus, the accuracy of the cultural diagnostic method turned out to be almost 3 times higher than the serological diagnostic method. Serological examination revealed 23% presence of Chlamydophila abortus in the samples, and the cultural diagnostic method revealed 60.9%. Unfortunately, in practice, the use of the cultural diagnostic method is associated with a time-delayed result, which is a disadvantage of this diagnostic method compared to the use of polymerase chain reaction. [ABSTRACT FROM AUTHOR]

Taking into account the wide range of physiological effects of vitamin D on the human body and the dependence of its synthesis in the skin on insolation, it is of undoubted interest to study vitamin D status in various population groups in the Russian North. The purpose of this article was to investigate vitamin D levels in the blood serum of people of various age groups living in Khanty-Mansiysk, Russia. Materials and methods. The research was conducted at the clinical and biochemical laboratory of the district clinical hospital in KhantyMansiysk during the autumn–winter–spring periods of 2021, 2022 and 2023. The study involved 3120 residents of Khanty-Mansiysk (61°N), including 493 children and adolescents, 798 young people, 887 middle-aged and 942 older adults. Their serum 25(OH)D levels were determined using enzyme immunoassay; the arithmetic mean (M), standard deviation and median (Me) as well as minimum and maximum values were calculated. The obtained data were compared with the reference values. Results. Vitamin D deficiency was established in all studied groups of the population of Khanty-Mansiysk regardless of their sex, being the most pronounced in children and adolescents due to intensive growth (M = 23.5 ng/ml, Me = 22.1 ng/ml) and in older adults (M = 22.5 ng/ml, Me = 21.7 ng/ml) due to significantly declined synthesis of cholecalciferol in the skin when exposed to ultraviolet radiation. There is a need for a widespread implementation of educational programmes informing healthcare workers and the population about the importance of vitamin D for metabolism, the prevalence of conditions related to vitamin D deficiency, and the measures for hypovitaminosis D prevention, such as consuming vitamin D-rich foods and taking vitamin D supplements in combination with longer sun exposure time in the south of Russia and abroad.

Earwax, a semi-solid substance, of both humans and non-human animals contains the stress hormone cortisol. Extraction of cortisol from earwax requires the use of organic solvents to eliminate interference factors. This study aimed to investigate the effects of organic solvents on the extraction efficiency of earwax cortisol from Hanwoo cattle. A factorial arrangement was used to determine the effects of solvent source (methyl alcohol [MA], diethyl ether [DE]) and assay diluent buffer (ADB) (0 and 1 mL) on earwax cortisol concentration (ECC) in Hanwoo cattle (parity 3, body weight = 462 kg). A 25 mg portion of the earwax sample was suspended in ADB, while another 25 mg portion was not suspended prior to extraction with MA and DE. The ECC was measured using a commercial enzyme-linked immunosorbent assay cortisol kit. The results indicated that cortisol can be extracted from earwax with or without ADB. The extraction yield of cortisol from earwax was higher with MA than with DE (p < 0.05). In conclusion, MA is the preferred solvent for cortisol extraction owing to its high extraction yield, simple extraction process, and relatively low solvent consumption. Moreover, earwax cortisol can be used as a biomarker to evaluate the degree of chronic stress in Hanwoo cattle.

Abstract Background The etiology of temporomandibular disorders (TMD) is multifactorial, involving a complex interplay of psychological and physiological factors. While salivary biomarkers, particularly cortisol, play an important role in TMD pathophysiology, evidence in the literature is still scarce and inconsistent. Hence, this study aims to evaluate the applicability of salivary cortisol as a potential biomarker for TMD in adults. Methods The Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) were used to accurately diagnose TMD patients. The study included adults, both male and female, aged 18–40 in the TMD group (n = 66) and non TMD participants (n = 66) matched for age and gender. Salivary samples were collected from participants at two time points: early and late morning. Cortisol levels in the samples were quantified using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using a one-way ANOVA to evaluate the correlations between cortisol levels and the study variables. Tukey’s post-hoc tests were applied to adjust for multiple comparisons. Results Salivary cortisol levels were significantly higher in the TMD group than in the corresponding controls (p = 0.034). In the TMD group, the mean cortisol levels early in the morning were 29.95 ± 75.05 (0-398.64), while the late morning levels were recorded as 4.87 ± 3.96 (0-17.13). In the control group, the mean cortisol levels early in the morning were 10.98 ± 16.83 (2.16–92.90), and late morning mean cortisol levels were 6.15 ± 6.13 (0-20.42). This indicates that the early morning levels of cortisol are higher in TMD patients (p = 0.046). In the subgroup analysis of the TMD, the mean salivary cortisol levels recorded were highest at 82.49 ± 124.34 (8.32-398.64) in patients having disc displacement without reduction with limited mouth opening. Furthermore, the mean salivary cortisol levels in the early morning were statistically higher (84.83 ± 132.80) in males compared to females (9.36 ± 9.01) (p = 0.008) with TMD. Conclusion The result of this study suggests that salivary cortisol could be a potential biomarker for a specific TMD subtype (disc displacement without reduction with limited mouth opening). However, further studies are needed to better understand the role of cortisol biomarker in the underlying pathogenesis of TMD.

Monitoring of humoral immunity in 3–4 year-old anti-pertussis vaccinated children is one of the approaches for pertussis epidemiological surveillance. It’s a predictive indicator for comprehensive assessment of morbidity in the region. The agglutination reaction (RA) used earlier has been displaced with enzyme immunoassay (ELISA). So, two questions arose: regarding i) a relevance of using ELISA in whole-cell vaccinated individuals, and ii) correctness of large-scale data interpretation without applying a unified IgG reference value. Study aims were to assess humoral anti-pertussis population immunity by ELISA, to determine features of humoral protection in vaccinated subjects after using various vaccine types, to identify age features, to make recommendations for evaluating seromonitoring data. Blood serum samples from anti-pertussis vaccinated children in Samara region (n = 1729) were examined in 2016–2020; children were divided into 4 age groups ( 3, 3–4, 5–7, ≥ 7.1 years) and into 3 type vaccines groups: cellular, acellular and combined. We used two ELISA kits, which are designed to assess post-vaccination immunity, according to the manufacturer’s instructions, quantifying IgG againstB. pertussis, which differ in units and reference values. Statistical analysis was performed using StatTech program v.2.6.2. The largest and smallest percentage of seronegative children was in the cellular-vaccinated and acellular-vaccinated group, respectively. Seronegative percentage in the “indicator group” reaches 55.2% being significantly higher than acceptable 10.0%-level, and differs from the literature data (30–40%), which were obtained mainly with RA. This may suggest about an incorrect interpretation of post-vaccination immunity data evaluation strictly according to manufacturers’ reference criteria, especially for whole-cell vaccinated subjects. A method for calculating the seronegative percentage is proposed that relies on the prognostic negative data level as a correction factor. The required values were obtained during comparison studies of random serum samples from 3–4-year-old children (n = 70) using two ELISA kits (“RIDASCREEN®Bordetella”, “SeroPertussis™ IgG”) and intra-laboratory comparison panel. It should be given preference for sets with a greater prognostic negative data level, if population seronegative percentage is assessed.

Introduction. Hemorrhagic fever with renal syndrome (HFRS) is the most common zoonotic human viral disease in the Russian Federation. More than 98% of the HFRS cases are caused by Puumala orthohantavirus (PUU). Effective serological tests are required for laboratory diagnosis of HFRS. Objective. Construction of an enzyme immunoassay (ELISA) test system for detection of specific antibodies using standard antigen in the form of highly purified inactivated PUU virus as immunosorbent. Materials and methods. Preparation of PUU virus antigen, designing the ELISA for detection of specific antibodies, developing parameters of the ELISA system, parallel titration of HFRS patients sera by fluorescent antibody technique (FAT) and the new ELISA. Results and discussion. For the first time, ELISA based on purified inactivated PUU virus as standard antigen directly absorbed onto immunoplate was developed. Parallel titration of 50 samples from HFRS patients blood sera using FAT and the developed ELISA showed high sensitivity and specificity of this ELISA, with 100% concordance of testing results and significant level of correlation between the titers of specific antibodies in the two assays. Conclusion. The ELISA based on purified inactivated PUU virus as an immunosorbent can be effectively used for HFRS serological diagnosis and for mass seroepidemiological studies.

Steroid hormones, especially glucocorticoids (GCs), are widely used to assess physiological responses to stressors. As steroid hormones are heavily metabolised prior to excretion, it is essential to validate enzyme immunoassays (EIAs) for measuring faecal glucocorticoid metabolites (fGCMs). Although problems with unvalidated assays have been raised repeatedly, their use persists widely.Lea et al. (2017) used an unvalidated corticosterone assay (CJM006) to relate fGCM concentrations to habitat quality, demography and population performance in the Cape mountain zebra (Equus zebra zebra). Here, we revisit their findings and evaluate the validity of their conclusions using a validated EIA. First, we evaluate the biological sensitivity of six EIAs (three group‐specific metabolite assays and three corticosterone assays, including CJM006) through a biological validation experiment (translocation) for two sub‐species of mountain zebra, Cape mountain and Hartmann's mountain zebra (E. z. hartmannae). Second, we reanalyse the faecal extracts from Lea et al. (2017) using a validated EIA.fGCM concentrations consistently increased following translocation, when using two 11‐oxoaetiocholanolone (lab codes: 72T and 72a) and an 11ß‐hydroxyaetiocholanolone (69a) EIA, but did not with three different corticosterone EIAs. All corticosterone EIAs (including CJM006) failed to detect an increase in fGCMs within the critical 48–72‐h period post translocation. Therefore, the CJM006 EIA utilised in Lea et al. (2017) does not sensitively measure hypothalamic–pituitary–adrenal (HPA) axis activity in CMZ faeces.Using a validated assay (72T), fGCM concentrations were no longer associated with adult sex ratio or habitat quality (measured by grassiness) and these variables were dropped from predictive models. fGCM concentrations now varied between seasons and were negatively associated with female fecundity (foal:mare ratio). Consequently, we can conclude that the results of the previous study are unreliable.We introduce the terms "insensitive" and "sub‐optimal" to categorise assays that are tested but fail validation, and assays that are comparatively poor at detecting relevant hormone changes, respectively. We discuss how both "insensitive" and "sub‐optimal" assays could lead to incorrect inferences about population stressors and counterproductive conservation recommendations. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

The eastern small eyed snake (Cryptophis nigrescens; CN) is an uncommon cause of snakebite in Australia despite the widespread distribution of the snake along the east coast of Australia. Diagnosis of envenomation relies on identification of the snake which is often not possible with animal snakebite cases. This study examined the immunoreactivity profile of CN venom towards specific rabbit IgG made against the medically relevant snake venom immunotypes found in Australia (tiger, brown, black, death adder and taipan). A simultaneous sandwich ELISA format was used to quantify CN venom binding to venom specific Protein A purified rabbit IgG. The binding profiles demonstrated weak binding of CN venom to rabbit IgG made against both tiger (N. scutatus) and black snake (P. australis) venoms with approximately 0.19% and 0.069% cross reactivity, respectively. However, the concentration of venom likely to be present in the urine of CN envenomed patients and the low cross reactivity suggest that envenomed veterinary patients are unlikely to be detected in the commercial snake venom detection kit. It is possible that CN envenomation is more common but may be underdiagnosed where snake venom antigen detection is relied upon solely. Serum biochemical abnormalities also overlap with other snake species found in the same geographical area. In respect of antivenom therapy, administration of tiger snake antivenom is supported by the binding data, but due to the low cross reactivity multiple vials may be required. Limited clinical evidence also supports the efficacy of tiger snake antivenom for envenomation by CN. [ABSTRACT FROM AUTHOR]

Introduction Winter warfare training (WWT) is a critical component of military training that trains warfighters to operate effectively in extreme environments impacted by snow and mountainous terrain. These environmental factors can exacerbate the disruption to the hormone milieu associated with operating in multi-stressor settings. To date, there is limited research on the physiological responses and adaptations that occur in elite military populations training in arduous environments. The purpose of this study was to quantify hormone responses and adaptations in operators throughout WWT. Materials and Methods Participants engaged in baseline laboratory metrics at their home station, Fort Carson, located in Colorado (CO) prior to WWT, for one week in Montana (MT) and one week in Alaska (AK). WWT periods were separated by approximately one month. Blood was collected upon wake at baseline (CO) and on the first and last day of WWT at each location (MT and AK). Plasma was analyzed for stress, metabolic, and growth-related hormones via enzyme-linked immunoassay (ELISA). Sleep quality was assessed via the Pittsburg Sleep Quality Index (PSQI) at baseline (CO) and on the first day of training in MT and AK. Cognitive function was evaluated using the Defense Automated Neurobehavioral Assessment (DANA) at baseline (CO) and on the first and last day of WWT in both MT and AK. Results Fourteen US Army operators in 10th Special Forces Group (SFG) Operational Detachment participated in winter warfare training (WWT; age: 31.5 years; 95%CI[28.1, 34.3]; height: 180.6 cm; 95%CI[177.3, 183.4]; weight: 87.4 kg.; 95%CI[80.6, 97.7]; body fat: 18.9%; 95%CI[13.7, 23.1]; male: n=13; female: n=1). Plasma adrenocorticotropic hormone (ACTH) levels increased from baseline (19.9 pg/mL; 95%CI[8.6, 24.2])  to pre-WWT (26.9 pg/mL; 95%CI [16.2, 37]; p=0.004), decreased from pre-  (26.9 pg/mL; 95%CI [16.2, 37]) to post-WWT in MT (22.3 pg/mL; 95% CI [8, 23.7]; p=0.004;), and increased from pre-  (25 pg/mL; 95%CI[ 28.4) to post-WWT (36.6 pg/mL; 95%CI [17.9, 48.9]) in AK (p=0.005). Plasma cortisol levels decreased from pre- (174 ng/mL; 95%CI[106.2, 233.6])  to post-WWT (94.5 ng/mL; 95%CI[54.8, 101.7]) in MT (p=0.001) and, conversely, increased from pre- (123.1 ng/mL; 95%CI[97.5, 143.9]) to post-WWT  (162.8 ng/mL; 95%CI[128, 216.7]) in AK (p<0.001). Alterations in growth-related hormones (insulin-like growth factor 1 [IGF-1], insulin-like growth factor binding protein 3 [IGFBP-3], and sex hormone binding globulin [SHBG]) were observed throughout WWT (p<0.05). The Total Testosterone / Cortisol ratio (TT / CORT; molar ratio) was lower pre-WWT in MT (0.04; 95%CI[0.01,0.04) compared to baseline in CO (0.07; 95%CI[0.04, 0.07]; p=0.042). Triiodothyronine (T3) levels increased from pre-  (101.7 ng/dL; 95%CI[93.7, 110.4]) to post-WWT  (117.8 ng/dL; 95%CI[105.1, 129.4]) in MT (p=0.042). No differences in sleep quality were reported between locations (CO, MT, and AK). Alterations in cognitive function were exhibited between locations and during WWT in both MT and AK (p<0.05). Conclusions Over the course of WWT, elite operators experienced alterations in stress, metabolic, and growth-related hormones, as well as cognitive performance. The increase in stress hormones (i.e. ACTH and cortisol) and reduction in cognitive performance following training in AK are suggestive of heightened physiological strain, despite similarities in physical workload, self-reported sleep quality, and access to nutrition. The variation in hormone levels documented between MT and AK may stem from differences in environmental factors, such as lower temperatures and harsh terrain. Further research is warranted to provide more information on the combined effects of military training in extreme environments on operator health and performance. [ABSTRACT FROM AUTHOR]

Background: Assessing the risk of measles outbreaks and identifying the susceptible parts of the population is essential to timely intervention. Infants between 6–12 months are increasingly susceptible to measles but evaluating the performance of high throughput enzyme immunoassays (ELISAs) in infants

Coccidioidomycosis is a fungal disease associated with soil exposure that frequently goes undiagnosed due at least in part to its nonspecific presentation and the lack of clinical suspicion by health care providers. Currently available diagnostics for coccidioidomycosis offer qualitative results that can suffer from low specificity, while semiquantitative assays are labor-intensive and complex and can require multiple days to complete. Furthermore, significant confusion exists regarding the optimal diagnostic algorithms and appropriate usage of available diagnostic tests. This review aims to inform clinical laboratorians and treating clinicians about the current diagnostic landscape, appropriate diagnostic strategies, and future diagnostic directions for coccidioidomycosis, which is expected to become more prevalent due to increased migration into areas of endemicity and climate changes.

Abstract Purpose The clinical significance of negative toxin enzyme immunoassays (EIA) for Clostridioides difficile infections (CDIs) is unclear. Our study aimed to investigate the significance of toxin EIA-negative in the diagnosis and prognosis of CDI. Methods All stool specimens submitted for C. difficile toxin EIA testing were cultured to isolate C. difficile. In-house PCR for tcdA, tcdB, cdtA, and cdtB genes were performed using C. difficile isolates. Stool specimens were tested with C. difficile toxins A and B using EIA kit (RIDASCREEN Clostridium difficile toxin A/B, R-Biopharm AG, Darmstadt, Germany). Characteristics and subsequent CDI episodes of toxin EIA-negative and -positive patients were compared. Results Among 190 C. difficile PCR-positive patients, 83 (43.7%) were toxin EIA-negative. Multivariate analysis revealed independent associations toxin EIA-negative results and shorter hospital stays (OR = 0.98, 95% CI 0.96–0.99, p = 0.013) and less high-risk antibiotic exposure in the preceding month (OR = 0.38, 95% CI 0.16–0.94, p = 0.035). Toxin EIA-negative patients displayed a significantly lower white blood cell count rate (11.0 vs. 35.4%, p

Nonylphenol is an aromatic organic compound that has an estrogen-like effect and has a negative effect on the human endocrine system. A method has been developed for the competitive determination of nonylphenol using magnetic particles, rabbit antiserum, nonylphenol conjugate with soybean trypsin inhibitor (STI) and biotin. The principle of the analysis is the formation of immune complexes on the surface of magnetite particles due to covalent immobilization of protein G through the oriented immobilization of polyclonal antibodies from rabbit serum during a competitive reaction between the free analyte (nonylphenol) and the bound one (as part of the nonylphenol-STI-biotin conjugate) for the binding sites of specific antibodies. The detection of formed immune complexes is proposed to be carried out using a streptavidin-polyperoxidase conjugate, which makes it possible to achieve a nine-fold gain in the level of the analytical signal. The developed ELISA using magnetite particles allows us to achieve a detection limit of nonylphenol at the level of 3.8 ng/mL, which is 14.5 times lower in comparison with the classic competitive ELISA (55 ng/mL). Based on the results of the experimental work, the optimized volume of the test sample was 500 μL, which makes it possible to concentrate low-contaminated samples by 17 times. [ABSTRACT FROM AUTHOR]

The pathogenicity of many bacteria, including Bacillus cereus and Staphylococcus aureus, depends on pore-forming toxins (PFTs), which cause the lysis of host cells by forming pores in the membranes of eukaryotic cells. Bioinformatic analysis revealed a region homologous to the Lys171-Gly250 sequence in hemolysin II (HlyII) from B. cereus in over 600 PFTs, which we designated as a "homologous peptide". Three β-barrel PFTs were used for a detailed comparative analysis. Two of them—HlyII and cytotoxin K2 (CytK2)—are synthesized in Bacillus cereus sensu lato; the third, S. aureus α-toxin (Hla), is the most investigated representative of the family. Protein modeling showed certain amino acids of the homologous peptide to be located on the surface of the monomeric forms of these β-barrel PFTs. We obtained monoclonal antibodies against both a cloned homologous peptide and a 14-membered synthetic peptide, DSFNTFYGNQLFMK, as part of the homologous peptide. The HlyII, CytK2, and Hla regions recognized by the obtained antibodies, as well as an antibody capable of suppressing the hemolytic activity of CytK2, were identified in the course of this work. Antibodies capable of recognizing PFTs of various origins can be useful tools for both identification and suppression of the cytolytic activity of PFTs. [ABSTRACT FROM AUTHOR]

Coccidiomycosis is a potentially life-threatening fungal infection endemic to certain regions of Argentina. The infection is caused by Coccidioides spp. and is primarily diagnosed by Coccidioides antibody (Ab) detection. Access to rapid, highly accurate diagnostic testing is critical to ensure prompt antifungal therapy. The sōna Coccidioides Ab Lateral Flow Assay (LFA) performs faster and requires less laboratory infrastructure and equipment compared with other Ab detection assays, potentially providing a substantial improvement for rapid case screening in coccidioidomycosis-endemic regions; however, validation of this test is needed. Thus, we aimed to evaluate the analytical performance of the sōna Coccidioides Ab (LFA) and compare agreement with anti-Coccidioides Ab detection assays. A total of 103 human sera specimens were tested, including 25 specimens from patients with coccidioidomycosis and 78 from patients without coccidioidomycosis. The sōna Coccidioides Ab Lateral Flow Assay (LFA) was performed with a sensitivity of 88%, and specificity and accuracy of 87%. Furthermore, the Coccidioides Ab LFA had good agreement with other anti-Coccidioides Ab detection assays. Our findings suggest the sōna Coccidioides Ab LFA has satisfactory performance and may be useful for diagnosing coccidioidomycosis in endemic regions. [ABSTRACT FROM AUTHOR]

Background Multistep laboratory testing is recommended for the diagnosis of Clostridioides difficile infection (CDI). The aim of this study was to present the impact of multistep CDI diagnostic testing in an academic hospital system and evaluate the toxin B gene polymerase chain reaction (PCR) cycle threshold (Ct) values of PCR-positive tests. Methods In October 2022, our system began reflex testing all PCR-positive stool samples with the C. DIFF QUIK CHEK COMPLETE (Techlab), an enzyme immunoassay–based test with results for the glutamate dehydrogenase antigen (GDH) and C difficile toxin A/B. Hospital-onset (HO) CDI and CDI antibiotic use before and after testing were tracked. Ct values were obtained from the Infectious Diseases Diagnostic Laboratory. Receiver operating curve analysis was used to examine the sensitivity and specificity for identifying GDH+/toxin+ and GDH−/toxin− at various Ct thresholds. Results The HO-CDI rate decreased from 0.352 cases per 1000 patient-days to 0.115 cases per 1000 patient-days post–reflex testing (P <.005). Anti-CDI antibiotics use decreased, but the decrease was not commensurate with CDI rates following reflex testing. PCR+/GDH+/toxin+ samples had a lower mean Ct value than PCR+/GDH–/toxin– samples (23.3 vs 33.5, P <.0001). A Ct value of 28.65 could distinguish between those 2 groups. Fifty-four percent of PCR+/GDH+/toxin− samples had a Ct value below that cut-off, suggesting the possibility of CDI with a negative toxin test. Conclusions Reflex testing for a laboratory diagnosis of CDI results in rapid, systemwide decreases in the rate of HO-CDI. Additional research is needed to distinguish CDI from C difficile colonization in patients with discordant testing. [ABSTRACT FROM AUTHOR]

Objective The serum squamous cell carcinoma antigen (SCCA) level is a well-known tumor marker for squamous cell carcinoma. In this study, we examined the impact of immunoglobulin (Ig)-bound macromolecular SCCA on serum SCCA levels measured by 2 different methods. Methods Seventy-five serum samples with an SCCA level >5.0 ng/mL as determined by a chemiluminescent immunoassay (CLIA) were also analyzed using a chemiluminescent enzyme immunoassay (CLEIA). The levels of IgG- and IgA-type anti-SCCA antibodies, which form immunoglobulins and macromolecules, respectively, were determined using an enzyme-linked immunosorbent assay. An absorption test was performed to confirm the presence of anti-SCCA antibodies. Results The correlation coefficient between the values measured by CLEIA and CLIA was 0.768. The ratio of SCCA levels measured by CLEIA to those measured by CLIA in 14 samples with IgG-type anti-SCCA antibodies was significantly lower than that in samples without these antibodies (P <.031). Absorption tests showed that SCCA levels measured by CLIA might be falsely high in samples with IgG-type anti-SCCA antibodies, probably due to reactions with SCCA1. Conclusion The level of SCCA as measured by CLIA and CLEIA methods correlate well, but the presence of SCCA antibodies can affect the results of the CLIA method. [ABSTRACT FROM AUTHOR]

Purpose: The clinical significance of negative toxin enzyme immunoassays (EIA) for Clostridioides difficile infections (CDIs) is unclear. Our study aimed to investigate the significance of toxin EIA-negative in the diagnosis and prognosis of CDI. Methods: All stool specimens submitted for C. difficile toxin EIA testing were cultured to isolate C. difficile. In-house PCR for tcdA, tcdB, cdtA, and cdtB genes were performed using C. difficile isolates. Stool specimens were tested with C. difficile toxins A and B using EIA kit (RIDASCREEN Clostridium difficile toxin A/B, R-Biopharm AG, Darmstadt, Germany). Characteristics and subsequent CDI episodes of toxin EIA-negative and -positive patients were compared. Results: Among 190 C. difficile PCR-positive patients, 83 (43.7%) were toxin EIA-negative. Multivariate analysis revealed independent associations toxin EIA-negative results and shorter hospital stays (OR = 0.98, 95% CI 0.96–0.99, p = 0.013) and less high-risk antibiotic exposure in the preceding month (OR = 0.38, 95% CI 0.16–0.94, p = 0.035). Toxin EIA-negative patients displayed a significantly lower white blood cell count rate (11.0 vs. 35.4%, p < 0.001). Among the 54 patients who were toxin EIA-negative and did not receive CDI treatment, three (5.6%) were diagnosed with CDI after 7–21 days without complication. Conclusion: Our study demonstrates that toxin EIA-negative patients had milder laboratory findings and no complications, despite not receiving treatment. Prolonged hospitalisation and exposure to high-risk antibiotics could potentially serve as markers for the development of toxin EIA-positive CDI. [ABSTRACT FROM AUTHOR]

We report on an analytical and biological validation of a commercial cortisol enzyme immunoassay (EIA) to measure glucocorticoids (GC) in feces of Geoffroy's spider monkeys (Ateles geoffroyi). Validation of endocrinological methods for each sample matrix and study species is crucial to establish that the methods produce reliable results. For the analytical validation of the EIA, we assessed parallelism, accuracy, and precision. We carried out a biological validation based on three well‐studied GC patterns with the following predictions: (1) increased fecal GC metabolite (fGCM) concentrations after veterinary intervention; (2) increased fGCM concentrations during early morning hours; and (3) higher fGCM concentrations during gestation than in other female reproductive states. For the first prediction, we sampled feces of two zoo‐housed females 2 days before, the day of, and 2 days after a veterinary intervention. For the second prediction, we analyzed 284 fecal samples collected from 12 wild males using a linear mixed model (LMM). For the third prediction, we analyzed 269 fecal samples of eight wild females using an LMM. Analytical validation revealed that the EIA showed parallelism, was accurate, and precise within each assay. However, there was elevated variation in between‐assay precision. The biological validation supported all predictions: (1) the two zoo‐housed females showed a substantial increase in fGCM concentrations 2.5 and 11 h after veterinary intervention; (2) there was a negative effect of sample collection time on fGCM concentrations (i.e., higher concentrations during early morning); (3) gestating females had significantly higher fGCM concentrations than lactating females. Thus, we analytically validated the commercial EIA and, despite between‐assay variation, we were able to find three biologically relevant GC signals in captive and wild settings, and in males and females. We are therefore confident that the method can be used to noninvasively address behavioral endocrinology questions in Geoffroy's spider monkeys. Research Highlights: Analytical validation of a commercial cortisol enzyme immunoassay for human serum to measure glucocorticoid metabolites in fecal sample extracts of Geoffroy's spider monkeys.Biological validation showing elevated fecal glucocorticoid metabolite levels in response to capture and anesthesia in captivity, during early morning hours in wild males, and in gestation compared with lactation and cycling in wild females.Noninvasive method available in home‐range country for future use in basic research of behavioral endocrinology and conservation studies in Geoffroy's spider monkey. [ABSTRACT FROM AUTHOR]

SCIENTIFIC RELEVANCE. The sanctions currently imposed on the Russian Federation requireusing Russian diagnostic products in vaccine quality assessments, as well as searching for alternative testing methods, in particular, serological ones.AIM. This study aimed to demonstrate the possibility of using alternative serological methods, including immunoenzymatic, direct agglutination, and indirect haemagglutination methods, for the identification of tularaemia and brucellosis vaccines in studies and quality assessments.MATERIALS AND METHODS. This study used the established pharmacopoeial reference standard (RS) for the bacterial suspension opacity of 10 international opacity units, the established pharmacopoeial RS for and two commercial batches of the live brucellosis vaccine (Brucella abortus 19 BA), and the established and candidate pharmacopoeial RSs for the live tularaemia vaccine (Francisella tularensis 15 NIIEG). These pharmacopoeial RSs were certified by the Scientific Centre for Expert Evaluation of Medicinal Products. Serological testing used Russian commercial diagnostic products, including dry diagnostic sera (polyvalent brucellosis and tularaemia sera for agglutination tests), a liquid erythrocytic diagnostic preparation of tularaemia immunoglobulin, and enzyme immunoassay (ELISA) diagnostic kits for detecting the causative agents for brucellosis and tularaemia. Statistical analysis involved using Microsoft Excel (P=0.95) for ELISA results and qualitative evaluation for the results of direct agglutination and indirect haemagglutination tests.RESULTS. All the tested batches demonstrated positive results. Live brucellosis vaccine batches showed positive results in the slide agglutination tests, while live tularaemia vaccine batches yielded positive results in the tube agglutination tests. All indirect haemagglutination tests showed haemagglutination in live tularaemia vaccine samples at the same concentration as positive control samples (6.25×10⁶ cells/mL). ELISA tests showed similar optical density values (D) for the two vaccines and positive control samples. Live tularaemia and brucellosis vaccines (undiluted, 1.0×10⁹ cells/mL) had D=2.133±0.273 and D=0.127±0.013, whereas the corresponding control samples had D=1.942±0.056 and D=0.123±0.007, respectively. The results reflected the presence of brucellosis or tularaemia microbes in the samples, confirming the identity of the vaccines.CONCLUSIONS. Serological immunoenzymatic, direct agglutination, and indirect haemagglu tination methods with Russian diagnostic products can be used to identify live brucellosis and tularaemia vaccines during quality assessment. The agglutination method with Russian diagnostic sera can be recommended as an alternative quality assessment method for the identification of live brucellosis and tularaemia vaccines, as this method offers time efficiency, simple visual evaluation of results, and low costs and relatively long shelf lives of diagnostic products. However, ELISA and indirect haemagglutination methods cannot be recommended for this purpose because of their labour-intensive and uneconomical nature. The results of this study may support the introduction of the agglutination method in the regulatory documents for live brucellosis and tularaemia vaccines (as an alternative method).

Abstract Background Although E. coli is generally a well-opted platform for the overproduction of recombinant antigens as heterologous proteins, the optimization of expression conditions to maximize the yield of functional proteins remains empirical. Herein, we developed an optimized E. coli (BL21)-based system for the overproduction of soluble immunoreactive HCV core/envelope proteins that were utilized to establish a novel immunoassay for discrimination of active HCV infection. Methods The core/E1-E2 genes were amplified and expressed in E. coli BL21 (DE3) in the absence/presence of glycylglycine. The antigenic performance of soluble proteins was assessed against 63 HCV-seronegative (Ab−) sera that included normal and interferent sera (HBV and/or chronic renal failure), and 383 HCV-seropositive (Ab+) samples that included viremic (chronic/relapsers) and recovered patients’ sera. The color intensity (OD450) and S/Co values were estimated. Results The integration of 0.1–0.4M glycylglycine in the growth media significantly enhanced the solubility/yield of recombinant core and envelope proteins by ~ 225 and 242 fold, respectively. This was reflected in their immunoreactivity and antigenic performance in the developed immunoassay, where the soluble core/E1/E2 antigen mixture showed 100% accuracy in identifying HCV viremic sera with a viral RNA load as low as 3800 IU/mL, without cross-reactivity against normal/interferent HCV-Ab−sera. The ideal S/Co threshold predicting active viremia (> 2.75) showed an AUC value of 0.9362 (95% CI: 0.9132 to 0.9593), with 87.64, 91.23% sensitivity and specificity, and 94.14, 82.11% positive and negative predictive values, respectively. The different panels of samples assayed with our EIA showed a good concordance with the viral loads and also significant correlations with the golden standards of HCV diagnosis in viremic patients. The performance of the EIA was not affected by the immunocompromised conditions or HBV co-infection. Conclusion The applicability of the proposed platform would extend beyond the reported approach, where glycylglycine, low inducer concentration and post-induction temperature, combined with the moderately-strong constitutive promoter enables the stable production of soluble/active proteins, even those with reported toxicity. Also, the newly developed immunoassay provides a cost-effective point-of-care diagnostic tool for active HCV viremia that could be useful in resource-limited settings.

It has been shown for the first time that humic preparations contain steroidal phytohormones— brassinosteroids (BS), the amount and composition of which vary depending on the source of raw materials and the method of its processing. It has been established that the qualitative and quantitative composition of BS correlates with the content of humic substances (humic acids). By the example of humic preparations obtained from sapropel, it was shown that alkaline treatment leads to the release of a noticeable amount of BS present in the form of conjugates. These results indicate that BS are important components of humic preparations and undoubtedly contribute to a wide range of physiological effects of these agricultural preparations. [ABSTRACT FROM AUTHOR]

Background Standalone nucleic acid amplification tests (NAATs) are frequently used to diagnose Clostridioides difficile infections (CDI), although they may be unable to distinguish colonization from disease. A 2-stage algorithm pairing NAATs with toxin immunoassays (Toxin) may improve specificity. We evaluated clinical outcomes of patients who were NAAT+/Toxin+ versus NAAT+/Toxin− and treated versus untreated NAAT+/Toxin− cases through systematic review and meta-analysis. Methods We searched EMBASE and MEDLINE from inception to April 1, 2023 for articles comparing CDI outcomes among symptomatic patients tested by NAAT and Toxin tests. The risk differences (RD) of all-cause mortality and CDI recurrence were computed by random effects meta-analysis between patients who were NAAT+/Toxin+ and NAAT+/Toxin−, as well as between patients who were NAAT+/Toxin− and treated or untreated. Results Twenty-six observational studies comprising 12 737 patients were included. The 30-day all-cause mortality was not significantly different between those who were NAAT+/Toxin+ (8.4%) and NAAT+/Toxin− (6.7%) (RD = 0.41%, 95% confidence interval [CI] = −.67, 1.49). Recurrence at 60 days was significantly higher among patients who were NAAT+/Toxin+ (19.8%) versus NAAT+/Toxin− (11.0%) (RD = 7.65%, 95% CI = 4.60, 10.71). Among treated compared to untreated NAAT+/Toxin- cases, the all-cause 30-day mortalities were 5.0% and 12.7%, respectively (RD = −7.45%, 95% CI = −12.29, −2.60), but 60-day recurrence was not significantly different (11.6% vs 7.0%, respectively; RD = 5.25%, 95% CI −1.71, 12.22). Conclusions Treatment of patients who were NAAT+/Toxin− was associated with reduced all-cause mortality but not recurrence. Although subject to the inherent limitations of observational studies, these results suggest that some patients who are NAAT+/Toxin− may benefit from treatment. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Almaty Technological University is the property of Almaty Technological University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Faecal glucocorticoid metabolites (FGMs) have gained relevance in ecological studies and population monitoring, allowing non-invasive remote sampling without the need to capture and handle animals. Enzyme immunoassays (EIAs) and radioimmunoassays (RIAs) are commonly used for FGM determination. Although these methods should be validated for each species, non-validated tests are still widely used to determine FGMs in wildlife. Near-infrared reflectance spectroscopy (NIRS) is a predictive method requiring calibration against reference methods used to assess FGMs in wildlife. EIAs and RIAs have been utilized to determine FGMs in chamois (Rupicapra spp.), a medium-sized mountain ungulate.This study aims to assess the potential of NIRS to determine FGMs and to evaluate the correlation among analytical methods used to determine FGMs in chamois. Faecal samples from 125 Alpine chamois (Rupicapra rupicapra rupicapra) and 125 Pyrenean chamois (R. pyrenaica pyrenaica) were collected from the field, frozen at − 20 °C, lyophilized, grounded, and scanned using a NIRSystems 5000 monochromator over a 1108–2492 nm wavelength. After this non-destructive NIRS analysis, FGMs were extracted and analysed using four immunoassays previously used in chamois studies: a 125-I-corticosterone RIA, a cortisol EIA, and two 11-oxoetiocholanolone EIAs (72a and 72T). Only the 11-oxoetiocholanolone 72T EIA has been validated for Alpine chamois. NIRS predictions were calibrated and cross-validated for each of the four immunoassays. The correlation among the four immunoassays was assessed using Spearman's rank.The coefficient of determination for NIRS calibration (R²) values ranged from 0.37 to 0.75, and the ratio of performance to deviation values from 1.2 to 1.6. Therefore, NIRS could not predict FGM concentration in chamois faeces. This could be due to the complexity and variability of the FGM detected by the immunoassays as reference methods, and by potential interference of other compounds in the faecal matrix. The correlation among the immunoassays was low overall.As a conclusion, NIRS cannot be recommended for measuring FGMs in chamois. The low correlation among the immunoassays used for FGM determination raises concern about the reliability of previous studies using non-validated methods in chamois. Only biologically validated tests should be used to assess FGMs to avoid incorrect inferences about biological responses in physiological, conservational, and ecological studies or population monitoring. These conclusions are applicable beyond the species studied here.

Introduction. Serum IgG measurement is used in the diagnosis and monitoring of many diseases. Therefore, it seems important to study the impact of coronavirus pandemic on IgG levels in population and the role of this parameter in COVID-19. The aim of this study was to compare mean IgG levels in sera obtained from 31 hospitalized COVID-19 patients, 30 healthy donors before pandemic and 34 donors vaccinated with «Sputnik V» (have not had COVID-19). Materials and methods. Total IgG was quantitated by two enzyme immunoassays (EIAs): «IgG Total-ELISA-BEST» kit certificated in Russia and homemade competitive EIA utilizing bispecific monoclonal antibodies (bsAbs) against human IgG (HIgG) and horseradish peroxidase (HRP). Results and discussion. The groups did not show differences in IgG levels (regardless of sex) with both methods giving comparable results. However, «IgG Total-ELISA-BEST» kit revealed statistically significant differences in mean serum IgG levels in subgroups of male patients depending on the levels of antibodies to viral RBD-antigen: below and above 400 BAU/ml. In the first subgroup (10 men) the mean serum IgG content was 14.3 ± 4.1 mg/mL, while in the second (6 men) — 6.9 ± 2.7 mg/mL. Conclusion. Sera obtained before pandemic contained the same mean IgG concentrations as sera from donors vaccinated with «Sputnik V» and COVID-19 patients. The relatively decreased mean IgG concentration was found only in COVID-19 male patients with anti-RBD antibodies levels above 400 BAU/ml. In light of literature data on association of decreased serum IgG with COVID-19 severity, it would be reasonable to further compare larger groups, taking into account clinical differences. The possibility of using bsAbs for human Ig measurements by competitive EIA was demonstrated.

Timely diagnosis and prevention of cardiovascular diseases is based on markers that detect changes in the athlete’s body at an early stage of disease. To implement this task, it is important to use novel laboratory techniques. We have carried out a comparative determination of immunological markers, specific antibodies to angiotensin, bradykinin, histamine, dopamine, serotonin and functional indicators of the cardiovascular system in athletes of various qualifications. The object of study included athletes of the Russian national teams who underwent an in-depth examination (IME) as part of medical survey. The participants were divided into groups depending on the sport arts and qualifications. Representatives of the group “Cyclic sports” included athletes without a category of 30 people and 29 qualified athletes (1st step, Candidate Master, Master of Sports, etc.). The “Combat sports” group consisted of 32 people without a category and 31 athletes with qualifications similar to those indicated above. Athletes of “Speed-strength” sports are represented by a group of 31 people without a category and 29 athletes with qualifications. The functional parameters of the cardiovascular system included analysis of heart rate, systolic blood pressure and diastolic blood pressure, which were compared with control values established for the athletes. The immunological indexes were determined in blood serum of the athletes and in subjects from control group by means of the solid-phase ELISA method using conjugated antigens of angiotensin, bradykinin, histamine, dopamine, serotonin for absorption on the plates. To compare the indexes, a control group of 30 people without cardiovascular symptoms was examined. Diastolic blood pressure for athletes of cyclic sports and combat sports was below the control values, and, for athletes of speed-strength sports, it was higher than in control group. A decrease in diastolic blood pressure for the above subgroup of athletes may indicate characteristic adaptive physiological changes in myocardium. The levels of immune-related indexes for serotonin, dopamine for the athletes of all groups corresponded to the control values. The only exception concerned qualified athletes of cyclic sports, which significantly differed from the normal values, compared to athletes without a sports category. Significantly high immunological parameters for histamine and angiotensin did not depend on the athlete’s qualification. In athletes involved in speed-strength sports, the levels of indexes for bradykinin did not differ from the normal values, and, for athletes in cyclic sports and combat sports, they significantly exceeded the norm.We have shown that different levels and types of sports activities affect the performance of cardiovascular system in their own way. The changes in immunological parameters reflect regulatory state of cardiovascular system. Accordingly, their simultaneous increase against the control may indicate participation in the development of cardiovascular diseases. The differences in their levels for athletes involved in combat sports, cyclic and speed-strength sporting activities show deeper changes in regulatory systems associated with duration and level of physical activities.

The development of COVID-19 is accompanied by involvement of various cytokines in pathological process. Their change depends on age, concomitant pathology and some other factors that have not been sufficiently studied in elderly patients with coronary heart disease (CHD). The content of cytokines in blood plasma of patients aged 60-74 in the early period of recovery from COVID-19 also remains unknown. The aim of our study was to determine the content of systemic cytokines in elderly patients with coronary heart disease at the early stages of recovery after COVID-19. The patients aged 60-74 with CHD who had COVID-19 of moderate severity grade (n = 40) made up the main group. in which the study of The cytokine levels were studied in blood plasma 3 to 4 weeks after recovery. The control group consisted of 38 elderly patients with coronary heart disease and negative tests for COVID-19. of cytokines in both groups were determined by means of flow cytometry with “Becton Dickinson FACS Canto 2 (USA)” machine using appropriate reagent sets. We have found that, in elderly patients with CHD at 3-4 weeks after recovery from COVID-19, if compared with elderly patients with CHD without COVID-19, the content of IL-6 in blood plasma was increased to higher degree and at statistically significant difference, up to 32.9±2.3 pg/mL versus 6.5±0.7 pg/mL in the control group (p < 0.001). Excessive content in the main group was also detected for IL-17 in blood plasma which was 25.4±1.9 pg/mL at the early post-COVID-19 period, whereas it was 7.8±0.7 pg/mL (p < 0.001) in the age-matched patients with CHD only. TNFα and IFNγ levels were elevated among 60-74-year-old patients with CHD and COVID-19 at 3-4 weeks after recovery, being 128.6±2.7 pg/mL and 57.6±2.8 pg/mL, respectively. In control group, the concentration of these cytokines was significantly lower, i.e., 56.3±2.2 pg/mL and 25.9±1.7 pg/mL, respectively (p < 0.001). The changes in contents of other cytokines studied seemed less pronounced, and the level of IL-4 was not significantly different between the both groups. Hence, in elderly patients with CHD and COVID-19 at 3-4 weeks after recovery, IL-6, IL-17, TNFα and IFNγ are the most elevated cytokines at the systemic level.

Scientific relevance. The immunoglobulin A (IgA) impurity content in parenteral human immunoglobulins should be determined in accordance with the State Pharmacopoeia of the Russian Federation by kinetic nephelometry, radial immunodiffusion, or enzyme immunoassay (ELISA) with a reference standard. The International Standard (IS) for the content of IgA is certified using gravimetry and radial immunodiffusion. However, neither of the existing standards for the content of IgA in human immunoglobulins is currently certified using all three compendial methods. This prevents analysts from comparing test results obtained by different methods and may lead to an underestimation of the IgA content in human immunoglobulins.Aim. This study aimed to determine the procedure for the development, certification, and use of a pharmacopoeial reference standard (RS) for the content of IgA in human immunoglobulins.Materials and methods. The authors studied candidate RSs for the IgA content derived from human plasma for fractionation. The IgA content determination involved kinetic nephelometry, radial immunodiffusion, and ELISA, as well as commercial test kits and the IS. The authors quantified the IgA impurity in samples of commercial human immunoglobulins from various manufacturers. The data analysis involved descriptive statistics and variance analysis using Microsoft Excel and Statistica 10.Results. The authors established a pharmacopoeial standard with a certified IgA content of 1.98 mg/mL (expanded uncertainty, 0.44 mg/mL; coverage coefficient, k=2; confidence level, 95%) for IgA impurity quantification in human immunoglobulins by radial immunodiffusion and ELISA and that of 1.31–2.64 mg/mL (expanded uncertainty, 0.67 mg/mL; coverage ratio, k=3; confidence level, 99%) for intralaboratory quality control of IgA impurity quantification by kinetic nephelometry, radial immunodiffusion, and ELISA.Conclusions. The pharmacopoeial standard developed in the study has been included in the register of standards of the State Pharmacopoeia of the Russian Federation as the Reference Standard for the Content of Immunoglobulin Class A (IgA) (Registry No. 3.1.00454). The pharmacopoeial standard is intended for the standardisation of analytical methods for the-determination of the IgA impurity content in parenteral human immunoglobulins.

Scientific relevance. Anti-hepatitis B virus surface-antigen (HBsAg) immunoglobulins are used to prevent hepatitis B in adults and children after exposure and to treat mild to moderate acute viral hepatitis B. The clinical effectiveness of human immunoglobulin preparations is determined by their potency, which is assessed by the content of antibodies to hepatitis B virus surface antigen (anti-HBs antibodies). Currently, this assessment involves using immunoassay techniques, such as enzyme-linked immunosorbent assay (ELISA).Aim. This study examined several mathematical methods for analysing the experimental data obtained in ELISA-based anti-HBs antibody potency assays of human immunoglobulin preparations.Materials and methods. This study used the international standard for human anti-HBs immunoglobulin, two immunoglobulin preparations, and an ELISA test kit for the detection and quantification of anti-HBs antibodies in serum or plasma samples.Results. Using sandwich ELISA, the authors ascertained that the measured anti-HBs antibody concentration depended on the choice of calibration curve calculation method (i.e. manual analysis, parallel-line analysis using PARALINE software, linear regression, and 4-parameter logistic regression). The measured anti-HBs antibody concentrations varied by ± 19 IU/mL. According to the study results, an incorrectly selected method of data analysis can lead to an erroneous calculation of the analyte potency (concentration) in the test sample.Conclusions. The study demonstrated the need for improved mathematical methods for the evaluation of experimental data used to determine the anti-HBs antibody concentration in human immunoglobulin preparations. It is essential to switch from manual to automated calculation (for example, using a 4-parameter logistic model), taking into account the requirements for bioanalytical methods and the capabilities of the available equipment.

Scientific relevance. Currently, two types of methods are used to evaluate the potency of diphtheria and tetanus toxoids: the gold standard, which involves administering toxins to immunised animals, and serological methods, which involve quantifying protective antibodies in the serum of immunised animals. International validation studies of serological methods for assessing the potency of diphtheria and tetanus toxoids have resulted in revisions to the relevant chapters of the WHO Manual for Quality Control of Diphtheria, Tetanus and Pertussis Vaccines, as well as the European, Japanese, and several other pharmacopoeias. Consequently, some diphtheria and tetanus vaccine manufacturers have substituted the potency evaluation methods that require administering toxins to animals with alternative serological methods.Aim. The study aimed to assess the suitability, reproducibility, and feasibility of an alternative enzyme immunoassay method (ELISA) for assessing the potency of diphtheria and tetanus components of diphtheria, tetanus, and whole-cell pertussis (DTwP) vaccines and to determine the possibility of implementing this method in Russia using standards and reagents manufactured in the country.Materials and methods. The study used combined vaccines for diphtheria prophylaxis, diphtheria toxin, and reference vaccines. The potency of diphtheria and tetanus toxoids was determined in guinea pigs and mice by the pharmacopoeial lethal challenge method and an alternative ELISA method.Results. ELISA and lethal challenge methods demonstrated comparable results of potency determination: 230 IU/mL vs 264 IU/mL (diphtheria toxoid), 188 IU/mL vs 160 IU/mL (tetanus toxoid), respectively. As opposed to the lethal challenge, the serological method allows testing the efficacy of several antigens in the same animals. The study showed the possibility of using purified diphtheria and tetanus toxoids manufactured in Russia as coating antigens. The authors obtained comparable antibody titres for each animal, using plates coated with international standards and Russian-made diphtheria and tetanus toxoids.Conclusions. The authors demonstrated the possibility of using ELISA to determine the potency of diphtheria and tetanus toxoids in DTwP vaccines. Moreover, the study demonstrated the suitability of Russian purified diphtheria and tetanus toxoids as coating antigens. Researchers should continue working on enzyme immunoassay validation with a view to harmonising national and international methods for assessing the potency of diphtheria and tetanus toxoid vaccines, as these efforts will not only facilitate the registration of foreign vaccines in Russia but also accelerate the approval of Russian vaccines in other countries.

Abstract Oxytocin has traditionally been known for its physiological effects on muscle contraction associated with birth and lactation, but in the last years is widely used as a biomarker of “positive experiences” in psychology and behavior. Different types of samples have been used for oxytocin measurements with saliva samples having the particular advantage of an easy and non-stressful collection. However, the low concentration of oxytocin in saliva can represent a limitation for its use. For this reason, sensitive assays and even a previous sample treatment in some cases are required for saliva oxytocin quantification. In addition, the lack of standardized and generally agreed-upon approach to peripheral oxytocin measurement leads to large discrepancies between different laboratories, that use different sample treatment protocols and different assays. The main objectives of this review are to describe the current status of the use of saliva for oxytocin measurement, provide details of the different sample processing techniques that can be applied and inform about the analytical techniques and assays available in different animal species, and also in humans for comparative purposes. It is expected that this information can contribute to an increase in the knowledge about the measurements of oxytocin in saliva and to its wider use in the future.

The ex situ polar bear population in the United States is not sustainable. Current management practices include pairing mates at five to six years of age, but there is evidence that wild bears have sired cubs as early as two years old. Timing acquisition of sexual maturity in male polar bears could aid in determining when mates should be paired in ex situ populations. To address this, non-invasive fecal testosterone metabolite monitoring was evaluated as a technique for establishing age of sexual maturity in 14 male polar bears (aged 1–6.9 years) for 1–6 years. Testosterone metabolite concentrations were monitored throughout the year and evaluated by breeding season (Jan. 1 – May 21) compared to non-breeding season (May 22 – Dec. 31). Sexual maturity was defined as when testosterone metabolite concentrations were significantly greater during breeding season than non-breeding season and bi-weekly averages were elevated above baseline for a minimum of ten weeks during breeding season mimicking patterns in adult male polar bears. Testosterone metabolite concentrations ranged from 5.9 to 8342 ng/g. One bear met the defined criteria at two years of age and an additional three bears met criteria at three years old. Due to the sampling strategy and the requirements set forth by the definition of sexual maturity, there were confounding results. In two cases, bears determined to be sexually mature the year prior, were not categorized as mature the next year. Another bear demonstrated greater testosterone metabolite values during the non-breeding season than breeding season. This study demonstrated the variability in fecal testosterone metabolites in juvenile polar bears and results suggest that hormone assessment alone is not adequate for determining sexual maturity in ex situ male polar bears. However, if used in tandem with behavioral monitoring and semen collection, when possible, could aid breeding management decisions for ex situ populations.

Hemolysin II (HlyII)—one of the pathogenic factors of Bacillus cereus, a pore-forming β-barrel toxin—possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibition of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemolysis on the presence of proline at position 324 in HlyII outside the conformational antigenic determinant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, obtained via multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interactions between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled the identification of the amino acid residues N339 and K340, included in the conformational antigenic determinant recognized by HlyIIC-20. [ABSTRACT FROM AUTHOR]

The study was carried out on Panthera species (4 lions and 5 tigers) housed in Bannerghatta Biological Park, Bengaluru, and Sri Chamarajendra Zoological Gardens, Mysuru. Estrus behavior such as rolling, licking, nudging, chuffing, frequent urination, whitish discharge, vocalisation, lordosis and copulatory behavior such as vocalisation, allowed to be mounted, aggression towards male, mounting, and biting of the nape of female were exhibited both in lions and tigers which helped to identify the time of fecal sample collection. The study's main aim was to carry out noninvasive and noncontact pregnancy detection in these wild felids. Feces samples collected from the female animals on day 0 (day of mating), 30, 45, 60, 90, 100, and 10th day postpartum were analysed for 13, 14-dihydro-15-keto-PGF2a metabolite (PGFM) levels using Enzyme Immunoassay (EIA). A significant increase in PGFM concentrations was seen from day 45 to day 60 in pregnant lionesses and tigresses used to detect pregnancy. The PGFM levels from day 60 were significantly different between pregnant and nonpregnant lionesses and tigresses. The PGFM levels were found to rise from day 45 and drop to basal levels by day 60 in 2 tigresses which may be due to fetal resorption. This study effectively detected pregnancy in lionesses and tigresses between day 45 and day 60 but requires further investigation due to the paucity in literature about PGFM analysis in tigers and lions. [ABSTRACT FROM AUTHOR]

Samples of macrophyte algae were collected from an ecotope of Kandalaksha Gulf, the White Sea. These algae were selected from substrates, storm wracks and after long-term storage in the soil, and they were assessed by indirect competitive enzyme immunoassay for the presence of low-molecular-weight metabolites of micromycetes (mycotoxins). In Fucus distichus, F. serratus, F. vesiculosus, and Ascophyllum nodosum, the concentrations of all analyzed toxic substances from storm wracks were uniformly reduced compared to those in living thalli. For Laminaria digitata, Saccharina latissima, and Ahnfeltia plicata, in living individuals of which mycotoxins were not found or were extremely rare, cases of contamination with citrinin, mycophenolic acid, emodin and sterigmatocystin were found in storm wracks. After 12 months of burial in soil F. distichus samples had only 9 components out of 16 analyzed. In F. serratus, F. vesiculosus, A. nodosum, and Pelvetia canaliculata, only emodin, mycophenolic acid, alternariol, and ergot alkaloids were determined. [ABSTRACT FROM AUTHOR]

Background: A rapid and reliable diagnostic test is needed to reduce mortality through early diagnosis of invasive aspergillosis (IA) in patients with hematological malignancies. Objective: To evaluate the efficacy of serum and bronchoalveolar lavage (BAL) Aspergillus galactomannan lateral flow assay (GM-LFA) in IA diagnosis and determine the correlation of GM-LFA with GM enzyme immunoassay (GM-EIA) in patients with hematological malignancies. Methods: In this prospective multicenter study, we used serum and BAL fluid samples from patients with hematological malignancies and suspected IA and performed GM-LFA and GM-EIA. According to the EORTC/MSGERC criteria, patients were grouped as proven (n = 6), probable (n = 22), possible IA (n = 55), or no IA (n = 88). The performance of serum GM-LFA at 0.5 optical density index (ODI) and area under the curve (AUC) were calculated. Spearman's correlation analysis and kappa statistics were performed to determine the agreement between the tests. Results: GM-LFA showed an AUC of 0.832 in proven/probable IA (sensitivity [SEN], specificity [SPE], negative predictive value [NPV], and diagnostic accuracy were 75%, 100%, 92.6%, and 93.9%, respectively, at a 0.5 ODI) versus that in no IA. A moderate positive correlation was noted between the GM-LFA and GM-EIA scores (p = 0.01). The observed agreement between the tests at 0.5 ODI was almost perfect (p < 0.001). After excluding patients who received mold-active antifungal prophylaxis or treatment, the SEN, SPE, NPV, and diagnostic accuracy for proven/probable IA were 76.2%, 100%, 93.3%, and 94.5%, respectively. Conclusions: Serum GM-LFA demonstrated high discriminatory power and good diagnostic performance for IA in patients with hematological malignancies. [ABSTRACT FROM AUTHOR]

C1 inhibitor of serine proteases (C1-INH) performs a regulatory function in the complement system and vascular permeability. Deficiency of C1-INH leads to various forms of angioedema, including hereditary angioedema (HAE). The cause of HAE is a genetically determined violation of the synthesis of C1-INH. A decrease in the level of C1-INH to 50% relative to the norm leads to an increase in the production of bradykinin, which is the basis for the diagnosis of HAE. The development of affordable ELISA for the quantitative determination of C1-INH is a popular direction for clinicians. During the development of a new kit for quantitative determination of C1-INH, two mouse monoclonal antibodies (mAb) with different epitope specificities were obtained. On their basis, a sandwich-type ELISA was developed. The specificity of the obtained mAb's was confirmed using the medical device “Berinert”. To prepare calibrators, C1-INH was affinity purified from human blood plasma using a sorbent with immobilized mAbs. The identity of the C1-INH protein was confirmed by PAGE electrophoresis, immunoblotting, and mass spectrometry on MALDI-TOF/TOF UltrafleXtreme mass spectrometer. To assess the quality indicators of developed reagents kit, studies were carried out in accordance with GOST R 51352-2013 and TU 21.20.23-041-01967164-2022. Values of quality indicators: accuracy — 93.53%; measurement linearity interval — 22.00-176.07 ng/mL. Using the developed ELISA test system, we examined 28 blood sera from healthy donors and 7 blood sera from patients with confirmed HAE. In the same samples, the content of C1-INH was determined by turbidimetric method, using the "Diagnostic reagents for in vitro immunochemical studies of specific blood proteins. Model: C1-esterase inhibitor (C1 EsteraseInhibitor)" (Aptec, Belgium). The correlation coefficient was 0.94 (p < 0.05). It was found that the diagnostic sensitivity and specificity of the developed ELISA is 100%. As a result of the study, an original ELISA test system for the quantitative determination of C1-INH was developed "Reagent kit for enzyme-linked immunosorbent assay of human C1-inhibitor (C1-inh PS)".

Studies of real opportunities for physical skills of athletes sufficiently depend on their adaptive potential for increasing physical loads. Extreme physical and psychoemotional loads may lead to overwork and decreased physical ability in professional sportsmen. These adaptation processes are regulated by the main biochemical systems of the body. A special role belongs to the factors of humoral immunity, i.e., natural antibodies, which are a component of innate immunity. They circulate in blood of healthy persons in absence of obvious antigenic stimulation. Analytical techniques for measuring the level of natural antibodies that reflect the state of the system of endogenous bioregulators involved into the molecular mechanisms of adaptation process have been developed. An important role among them is played by the regulators of the opioid system β-endorphin and orphanin. The biochemical and immunological parameters were determined in 10 athletes active in figure skating (Master of Sports), whose average age was 16±0.4 years, and sport experience of 9±1 years. The duration of the study was divided into 5 stages and was 62 days. During the dynamic observations in the course of intensive training, no clear shifts in biochemical parameters were revealed towards adaptation stress and delayed recovery. The level of natural antibodies to orphanin and beta-endorphin was measured in the athletes blood serum by ELISA techique. It is found that each athlete is characterized by individual immune profile. At the initial stage of the examination, the level of antibodies to beta-endorphin was within normal ranges, except for its decrease in one athlete. The level of antibodies to orphanin in majority of cases was higher than normal, probably, due to inhibitory control of the pain signal. Further study in time dynamics revealed that the immunological parameters, natural antibodies to opioid peptides, change in accordance with the state of adaptation resources in the athletes. These indexes reflect psycho-emotional potential and pain tolerance threshold for athletes from the start of training and throughout the entire period. Therefore, from a prognostic point of view, it is important to monitor the content of natural antibodies to beta-endorphin and orphanin in athletes in the course of training. Such individual monitoring of the athlete’s immunological indices allows us to select a more effective, personal training program.

Introduction. Registration of regular epidemic outbreaks of mumps infection among young people, difficulties in the clinical diagnosis of the disease, the presence of specific clinical manifestations in adults with a severe course of infection make it necessary the improvement of the quality of diagnosis of the disease. The aim of the work is to develop a standard definition of a clinical case of mumps and to conduct a comparative assessment of the effectiveness of the diagnostic test systems used for mumps diagnostics based on the determination of their sensitivity and specificity. Materials and methods. To develop a standard definition of a clinical case of mumps based on ten combined clinical and laboratory parameters, a mathematical model was built using the logistic regression method. The study included 84 patients with mumps infection (main group) and 66 patients with non-epidemic sialoadenitis (comparison group). For a comparative evaluation of the diagnostic systems VectoParotitis-IgM, VectoParotitis-IgG, ELISA-Mumps-IgM, ELISA-Mumps-IgG, Anti-Mumps Virus ELISA-IgM, and Anti-Mumps Virus ELISA-IgG, a blood serum samples from patients (n = 41) with a diagnosis mumps? were tested in dynamics to assess their sensitivity, specificity, and diagnostic effectiveness. Results. The clinical symptom dry mouth increased the chance of diagnosing mumps by 13.7 times, the presence of bilateral lesions of the salivary glands by 6.5 times, the increase in the level of diastase in the urine by 3.2 times. The coefficient of determination R2 was 61.1; the sensitivity was 93% and the specificity 67%; p 0.001. Diagnostic effectiveness of Anti-Mumps Virus ELISA-IgG test system was 84.4%, ELISA-Mumps-IgG 87.2%, and "VectoParotitis-IgG" 88.5%. Conclusions. Based on the combination of following clinical symptoms observed simultaneously in patient, of the general weakness, dry mouth, bilateral lesions of the salivary glands, an increase in the level of diastase in the urine, it is possible to establish a clinical diagnosis of mumps with a probability of 81%. The highest diagnostic effectiveness of domestically manufactured test systems for quantitation of IgG antibodies to mumps virus in sick people has been demonstrted.

Simple Summary: Glucocorticoids (GCs) play a vital role in the body's response to stress. Therefore, scientists use GC level measurements as a valuable tool to understand the well-being of animals in behavioral, conservation, and welfare research. GC levels can be measured from different sample types, one of them being saliva. Saliva samples are especially useful because they can be collected rather easily and in a non-invasive manner. The aim of this study is to validate a method to measure salivary GCs, i.e., an enzyme immunoassay, in two monkey species: capuchin monkeys and squirrel monkeys. Individuals of both species were trained to provide saliva samples. Using high-pressure liquid chromatography, we confirmed that two main GCs, cortisol, and cortisone, are present in the saliva of these species and that they can be detected by the enzyme immunoassay. We further verified the biological validity of the assay in the two species by showing that it can track the decrease in cortisol levels throughout the day, which is typical in these monkeys. Our findings support the usefulness of the enzyme immunoassay to measure salivary cortisol levels in these non-human primate species and highlight its potential as a valid tool in research and welfare assessment. Glucocorticoids (GCs) are mammalian steroid hormones involved in a variety of physiological processes, including metabolism, the immune response, and cardiovascular functions. Due to their link to the physiological stress response, GC measurement is a valuable tool for conservation and welfare assessment in animal populations. GC levels can be measured from different matrices, such as urine and feces. Moreover, especially in captive settings, measuring GCs from saliva samples proved particularly useful as those samples can be collected non-invasively and easily from trained animals. Salivary GC levels can be measured using a variety of analytical methods, such as enzyme immunoassays. However, it is crucial to validate the analytical method for each specific application and species when using a new matrix. Using high-pressure liquid chromatography and a cortisol enzyme immunoassay, we show that the main glucocorticoids secreted in the saliva of squirrel monkeys and brown capuchin monkeys are cortisol and cortisone. Our biological validation found the expected salivary cortisol level to decline throughout the day. Our findings support the reliability of salivary cortisol measurements and their potential to be used as a valid tool in research and welfare assessment for these non-human primates. [ABSTRACT FROM AUTHOR]