Polymorphism of Genes Encoding Inflammatory Interleukins and the Risk of Anterior Cruciate Ligament Injury: A Systematic Review and Meta-Analysis.

Sport injuries, including the anterior crucial ligament rupture (ACLR) seem to be related to complex genetic backgrounds, including the genes responsible for inflammatory response. This review and meta-analysis investigated the contribution of the polymorphisms of genes encoding inflammatory cytokines and their receptors to the risk of ACLR. The scientific databases Science Direct, EBSCO host, Scopus, PubMed, and Google Scholar were screened (completed on 14 June 2023) according to the established inclusion/exclusion criteria (only fully accessible, original, human case-control studies written in English concerning the effect of interleukin genes' polymorphisms on the occurrence of ACL injury were included) and statistical meta-analysis using R version 4.0.3 was performed. The PRISMA methodology was used to review articles. The review protocol was registered under the number CRD42024514316 in the Prospero database. Eighty-nine studies were identified and narrowed down to three original case-control studies used for the meta-analysis. The studies analyzed Polish, South African, and Swedish cohorts, altogether 1282 participants. The candidate polymorphisms indicated in the studies involved IL6 rs1800795, IL6R rs2228145 and IL1B rs16944. The systematic review showed the relationships between IL6 rs1800795 polymorphism and ACLR in the Polish subpopulation, and IL6R rs2228145 and IL1B rs16944 in the South African subpopulations. The meta-analysis revealed that the IL6 rs1800795 CG genotype was over-represented (OR = 1.30, 95% CI 1.02-1.66), while the CC genotype was under-represented (OR = 0.75, 95% CI 0.54-1.03) in ACLR subjects, but no significant impact of IL6R rs2228145 was shown. Additionally, a tendency of the IL1B rs16944 CT genotype to be protective (OR 0.89, 95% CI 0.70-1.14), while the TT to be a risk genotype (OR 1.19, 95% CI 0.84-1.68) was observed. Thus, the relationship between the interleukin receptor IL6R rs2228145 and ACLR risk was not confirmed. However, the impact of genes coding pleiotropic IL6 rs1800795 on the incidences of ACLR was clear and the effect of pro-inflammatory IL1B rs16944 was possible.

Proteome and phospholipidome interrelationship of synovial fluid-derived extracellular vesicles in equine osteoarthritis: An exploratory 'multi-omics' study to identify composite biomarkers.

Osteoarthritis causes progressive joint deterioration, severe morbidity, and reduced mobility in both humans and horses. Currently, osteoarthritis is diagnosed at late stages through clinical examination and radiographic imaging, hence it is challenging to address and provide timely therapeutic interventions to slow disease progression or ameliorate symptoms. Extracellular vesicles are cell-derived vesicles that play a key role in cell-to-cell communication and are potential sources for specific composite biomarker panel discovery. We here used a multi-omics strategy combining proteomics and phospholipidomics in an integral approach to identify composite biomarkers associated to purified extracellular vesicles from synovial fluid of healthy, mildly and severely osteoarthritic equine joints. Although the number of extracellular vesicles was unaffected by osteoarthritis, proteome profiling of extracellular vesicles by mass spectrometry identified 40 differentially expressed proteins (non-adjusted p < 0.05) in osteoarthritic joints associated with 7 significant canonical pathways in osteoarthritis. Moreover, pathway analysis unveiled changes in disease and molecular functions during osteoarthritis development. Phospholipidome profiling by mass spectrometry showed a relative increase in sphingomyelin and a decrease in phosphatidylcholine, phosphatidylinositol, and phosphatidylserine in extracellular vesicles derived from osteoarthritic joints compared to healthy joints. Unsupervised data integration revealed positive correlations between the proteome and the phospholipidome. Comprehensive analysis showed that some phospholipids and their related proteins increased as the severity of osteoarthritis progressed, while others decreased or remained stable. Altogether our data show interrelationships between synovial fluid extracellular vesicle-associated phospholipids and proteins responding to osteoarthritis pathology and which could be explored as potential composite diagnostic biomarkers of disease.

Molecular insights into the lipid-carbohydrates metabolism switch under the endurance effort in Arabian horses.

BACKGROUND: Recent studies have shown that in Arabian horse muscle, long-term exercise-induced expression of genes related to fatty acid degradation and the downregulation of genes belonging to the glycolysis/gluconeogenesis and insulin signalling pathways. Long-lasting physical exertion may trigger the metabolism to switch the main energy source from carbohydrates to lipids due to higher caloric content. OBJECTIVES: To describe the metabolism adaptation at the whole transcriptome of blood to endurance effort in Arabian horses. STUDY DESIGN: In vivo experiment. METHODS: Venous blood samples from 10 Arabian horses were taken before and after a 120 km long endurance ride to isolate the RNA and perform the high-throughput NGS transcriptome sequencing. RESULTS: The results, including KEGG (Kyoto Encyclopaedia of Genes and Genomes) and GO (Gene Ontology) analyses, allowed us to describe the most significantly upregulated-ARV1, DGAT2, LIPE, APOA2, MOGAT1, MOGAT2, GYS1, GYS2 and downregulated-ACACA, ACACB, FADS1, FADS2 genes involved in carbohydrate and lipid metabolism. Also, the increased expression of RAF1, KRAS and NRAS genes involved in the Insulin pathway and PI3K-Akt was shown. MAIN LIMITATIONS: Limited sample size, Arabians used for endurance racing were not compared to Arabians from other equestrian disciplines. CONCLUSIONS: This general insight into the processes described supports the thesis of the lipid-carbohydrates metabolism switch in endurance Arabian horses and provides the basis for further research.

Metabolomic analysis indicated changes in triacylglycerols' levels as a result of training in Whippet dogs.

Regular physical effort produces metabolic changes manifested as adaptation to exercise and increasing performance. In humans these changes have been characterized at metabolome level as depending on the discipline. However, all sports involve some level of changes in protein, carbohydrate and lipid metabolism. Recently, also performance horses have been subjected to metabolic analyses, but similar studies were lacking in sports dogs. In this study we performed the metabolomic analysis in plasma of Whippet dogs regularly trained and competing in coursing events, and untrained dogs of the same breed, fed with the same diet. We have also compared the hematological and blood biochemical results in these two groups of dogs. Basic blood tests indicated that enzymes related to lipid metabolism (lipase and gamma-glutamyltransferase) differed considerably between the groups. Metabolomic analysis of plasma confirmed the metabolic shift expressed as the differences in triacylglycerols levels between training and non-training dogs, aimed at improving the use of fatty acids as a source of energy during exertion. Surprisingly, other classes of metabolites were only hardly changed when comparing training and non-training Whippets.

Exercise-induced Haematological and Blood Lactate Changes in Whippets Training for Lure Coursing.

Introduction: Whippets are traditionally trained to compete in lure coursing. While in humans and horses, training is routinely monitored by special tests, this is not carried out in the training of whippets. The aim of this study was to check if laboratory tests designed for racehorses could be useful in monitoring whippets training for lure coursing. Material and Methods: Blood samples were taken from 14 whippets at several time points: before exercise (including warm-up), immediately after, 15 min after and 30 min after exercise sessions of straight 400 m runs (T) and coursing (C). Routine haematological values and lactate concentrations (LA) were measured. Results: White blood cell count, red blood cell count, haemoglobin concentration and haematocrit increased significantly in both types of exertion, and no differences between the types were observed. The LA measured immediately after the run were increased, but there was no significant difference between the types of session (T and C). After both types of activity, LA decreased within 30 min post run by 9-11 mmol/L. Lactate concentrations were significantly higher 30 min after the T sessions than after the C sessions. Conclusion: The results confirmed that typical exercise-induced changes occurred in whippets training for lure coursing; however, the scale of changes was different to that in horses. The sampling scheme used in racehorses can be applied to whippets and can be useful as a laboratory tool for monitoring their training.

Less Typical Courses of Rhodococcus equi Infections in Foals.

This article aims to present several interesting and less typical courses of Rhodococcus equi infections in foals, collected during the 2019-2021 foaling seasons in some Polish studs. The study was conducted by the Division of Veterinary Epidemiology and Economics, Warsaw University of Life Sciences-SGGW, and concentrated on ultrasonographic contribution to diagnostics and treatment of the disease. Among many standard cases of rhodococcal pneumonia, some rare ones occurred. The aforementioned issues include the potential contribution of rhodococcal infection to a grave outcome in a prematurely born filly, lost as a yearling, so-called "extrapulmonary disorders" (EPD), a hypothesis of inherited immunodeficiency with grave outcome in a breeding dam line from one stud, and macrolide-induced anhidrosis. The main benefit of this report would be to supplement the general picture of clinical rhodococcosis.

The effect of IL10 gene polymorphism on obesity parameters in highly physically active young men.

The main aim of this study was to investigate the association between 5 polymorphisms of the interleukin 10 (IL10) gene and body composition parameters in physically active young men. A cohort of 131 young men was enrolled and the following IL10 single-nucleotide polymorphisms (SNPs) were analysed: rs1518111, rs1878672, rs3024496, rs3024498 and rs3024505. The subjects were divided into groups depending on obesity parameters: body mass index (BMI) and percentage of body fat tissue (fat %). Statistical analysis was conducted for alleles, genotypes and haplotypes, and an association between SNPs and body composition parameters was analysed using four genetic models: dominant, recessive, codominant and overdominant mode of inheritance (MOI). The only statistically significant result in polymorphisms was found for rs3024505 in the over-dominant model with BMI (p = 0.04) and with fat % (p = 0.02). The haplo.score function showed an association between BMI and CCGTA (respectively) haplotype in the additive model (score = -2.00, p = 0.04) and in the dominant model (score = -2.30, p = 0.02). The obtained results indicate a statistically significant contribution of selected IL10 polymorphisms in the regulation of body weight in physically active individuals.

MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

An increase in the percentage of monocytes with reduced HLA-DR expression and immunosuppressive properties has been reported in numerous human neoplastic diseases, including lymphoma. However, there are no analogous studies on phenotypical variations in the peripheral blood monocytes in dogs with lymphoma. The aim of this study was to determine the difference in the expression of the MHCII molecule on peripheral blood monocytes in dogs with lymphoma before any treatment (NRG) and in dogs that had previously received glucocorticoids (RG) in comparison to healthy dogs. Flow cytometry immunophenotyping of peripheral blood leukocytes was performed using canine-specific or cross-reactive antibodies against CD11b, CD14 and MHCII. In the blood of dogs with lymphoma (NRG and RG), compared to that of healthy ones, the MHCII+ and MHCII- monocytes ratio was changed due to an increase in the percentage of MHCII- monocytes. The number of MHCII- monocytes was significantly higher only in RG dogs compared to healthy ones, which might result from the release of these cells from the blood marginal pool due to the action of glucocorticoids. Our results encourage further studies to assess if changes in MHCII expression affect immune status in dogs with lymphoma.

Comparative Histology of C Thyrocytes in Four Domestic Animal Species: Dog, Pig, Horse, and Cattle.

The number, morphology, and distribution of C thyrocytes within the thyroid gland vary among species; however, studies in domestic animals are limited. In this study we compared the morphology, distribution pattern, and percentage of C thyrocytes in four domestic species: dogs, pigs, horses, and cattle. Eighty thyroid glands, 20 per species, were examined. C thyrocytes were visualized immunohistochemically with anti-calcitonin rabbit polyclonal antibody alone and combined with the periodic acid Schiff method to simultaneously visualize C thyrocytes with the basement membranes of thyroid follicles. C thyrocyte morphology varied considerably between species, from oval- (dogs) and spindle-shaped (pigs) to polymorphic (cattle and horses). Bovine C thyrocytes demonstrated cytoplasmic protrusion. C thyrocytes were located intrafolliculary (all species), epifollicularly (dogs, horses, cattle), or interfolicularly (cattle). Most porcine and bovine C thyrocytes existed individually whereas canine C thyrocytes usually formed clusters. In horses, they tended to form groups of various shapes and sizes or even rims encompassing whole follicles. In all species, the number of C thyrocyte profiles increased from the periphery to the central area of the thyroid lobe. The mean total fraction of C thyrocytes in the superficial, intermediate, and central areas were as follows: 2.55%, 8.43%, and 12.48% in dogs; 3.81%, 7.66%, and 10.79% in pigs; 1.55%, 7.44%, and 8.87% in horses; and 2.62%, 10.75%, and 12.96% in cattle. No statistical differences in the total number of C thyrocyte profiles were observed among species (8.87% in dogs, 8.58% in cattle, 7.98% in pigs, and 5.83% in horses). Our results indicated that the studied species displayed their own morphological characteristics and distribution pattern of C thyrocytes; however, total numbers of C thyrocyte profiles and their localization within the thyroid lobe are comparable.

The interactions between interleukin-1 family genes: IL1A, IL1B, IL1RN, and obesity parameters.

BACKGROUND: Obesity has been recognized as a worldwide growing problem, producing many pathologies including the promotion of "proinflammatory state." The etiology of human obesity is still only partially understood; however, the genetic background has been proved. Its nature is complex, and currently, it appears that the combined effects of the interactions among multiple genes should receive more attention. Due to the fact that obesity promotes proinflammatory conditions, in this study, we investigated the genetic polymorphism of IL-1 family genes in healthy people with normal and elevated body mass index (BMI) and fat %. RESULTS: The single-nucleotide polymorphisms (SNPs) within the IL1A -889C > T (rs1800587), IL1B + 3954 T > C (rs1143634), and IL1RN -87G > A (rs2234677) genes alone were associated neither with BMI nor fat % values in tested group. The associations between SNP-SNP interaction and BMI for the IL1B × IL1RN interactions were significant for dominant model (p = 0.02) and codominant model (p = 0.03). The same SNP-SNP interaction (IL1B × IL1RN) was associated also with fat % for codominant (p = 0.01) and recessive (p = 0.002) models. CONCLUSIONS: This study further confirmed that IL-1 family genes are involved in genetic background of obesity. It has been shown that interaction IL1B × IL1RN was associated with both BMI and fat % with rare T allele protecting form higher values. Thus, even if certain polymorphisms in single genes of IL-1 family cannot be defined as related to obesity in examined population, the genetic interrelationships should be analyzed.

IL-6 Polymorphisms Are Not Related to Obesity Parameters in Physically Active Young Men.

Interleukin 6 (IL-6) is a cytokine with both pro- and anti-inflammatory actions, but is also considered as a "metabolic hormone" involved in immune responses, affecting glucose, protein and lipid metabolism. It has been proposed to be related to obesity, but various results have been presented. Thus, in this study, the very homogenous population of young, male military professionals, living in the same conditions involving high physical activity, has been selected to avoid the influence of environmental factors. The subjects were divided into groups depending on the obesity parameters BMI (body mass index) and fat percentage (fat%), and the following IL-6 SNPs (Single Nucleotide Polymorphisms) were analyzed: rs1800795, rs1800796 and rs13306435. No relation was found between obesity parameters and IL-6 polymorphisms rs1800795, rs1800796 and rs13306435. It may be postulated that even if a genetic predisposition involves IL-6 genes, this effect in individuals with obesity of a low grade is minor, or can be avoided or at least markedly reduced by changes in lifestyle.

Gut Microbiota Manipulation in Foals-Naturopathic Diarrhea Management, or Unsubstantiated Folly?

Diarrhea in foals is a problem of significant clinical and economic consequence, and there are good reasons to believe microbiota manipulation can play an important role in its management. However, given the dynamic development of the foal microbiota and its importance in health and disease, any prophylactic or therapeutic efforts to alter its composition should be evidence based. The few clinical trials of probiotic preparations conducted in foals to date show underwhelming evidence of efficacy and a demonstrated potential to aggravate rather than mitigate diarrhea. Furthermore, recent studies have affirmed that variable but universally inadequate quality control of probiotics enables inadvertent administration of toxin-producing or otherwise pathogenic bacterial strains, as well as strains bearing transferrable antimicrobial resistance genes. Consequently, it seems advisable to approach probiotic therapy in particular with caution for the time being. While prebiotics show initial promise, an even greater scarcity of clinical trials makes it impossible to weigh the pros and cons of their use. Advancing technology will surely continue to enable more detailed and accurate mapping of the equine adult and juvenile microbiota and potentially elucidate the complexities of causation in dysbiosis and disease. In the meantime, fecal microbiota transplantation may be an attractive therapeutic shortcut, allowing practitioners to reconstruct a healthy microbiota even without fully understanding its constitution.

Serological Survey of Leptospira Infection in Arabian Horses in Poland.

Leptospirosis is one of the most common zoonotic infections worldwide, including in most livestock, some companion animals, horses, wildlife, and humans. Epidemiological estimation of its prevalence in all species is difficult due to the variety of clinical presentations and challenges regarding laboratory diagnosis. The purpose of this study was to measure the seroprevalence of leptospiral infection in Arabian horses kept in the largest breeding farms in Poland, representing over 15% of the Polish Arabian horse population. Leptospira antibodies were detected by MAT (cut-off 1:100) in 33.2% of serum samples (204 of 615 animals) (CI 95%: 29.6-37.0%), most frequently reacting with the serovar Grippotyphosa, similar to previous reports in populations of randomly selected horses. These results indicated high Leptospira seropositivity, thus, although any form of clinical leptospirosis is rare, it may be postulated that the leptospiral exposure is widespread.

Variations in haematological and biochemical parameters in healthy ponies.

BACKGROUND: Breed specific reference ranges for selected blood parameters are recommended for proper interpretation of blood tests, but there are only few reports dealing with ponies. The purpose of this study was to investigate if blood parameters differ among ponies' classes and to check if general normal values for equine species are applicable to ponies. RESULTS: All, except total protein concentration, biochemical parameter significantly (p < 0.05) differed among ponies' classes. The most pronounced difference was noted in blood lactate concentrations, higher (p < 0.001) in the smallest ponies (class A). In all groups of ponies muscle enzymes (aspartate aminotransferase and creatine kinase) and urea were high when compared to normal values for equine species, but triglycerides and creatinine were low. Blood lactate concentration was high in comparison with normal values for horses only in class A ponies'. CONCLUSIONS: In healthy ponies, blood lactate concentration significantly differs between height classes. Normal values for equine species should not be directly applied to interpret the lactate, triglycerides, aspartate aminotransferase and creatine kinase values in ponies.

Association between peroxisome proliferator-activated receptor-alpha, -delta and -gamma gene (PPARA, PPARD, PPARG) polymorphisms and overweight parameters in physically active men.

Peroxisome proliferator-activated receptors (PPARs) have unique functions in energy metabolism regulation but are also involved in regulation of the inflammatory process and obesity. The aim of this study was to analyse potential associations between polymorphisms of PPARA (rs1800206), PPARD (rs1053049; rs2267668) and PPARG (rs1801282) and overweight parameters. One hundred and sixty-six males, unrelated Caucasian military professionals, were recruited in the genetic case-control study conducted in the period 2016-2019. All the participants were aged 21-41 and had similar levels of physical activity. Body mass, height and body composition were measured. The participants were divided into two groups depending on their BMI (body mass index) and FMI (fat mass index). The control group consisted of people with BMI between 20.0 and 25.0 or FMI values ≤ 6, while the overweight group consisted of people with BMI of ≥ 25.0 or FMI values > 6. Genomic DNA was isolated from extracted buccal cells. All samples were genotyped using real-time polymerase chain reaction (real-time PCR). It was found that two polymorphisms rs2267668 and rs1053049 of the PPARD gene were significantly associated with BMI: SNP rs2267668 for the dominant (OR = 2.04, 95%CI 1.01-4.11, p-value = 0.04) model (A/G-G/G vs A/A). The likelihood of being overweight was over 2 times smaller for allele A. A relationship between the polymorphism of PPARG (rs1801282) and BMI was found for the overdominant (OR = 2.03, 95%CI 1.03-4.00, p-value = 0.04) model (C/G vs C/C-G/G). Significant associations were found in different models for PPARD, PPARG and PPARA genes with BMI. In SNP rs2267668 for the codominant genetic model (G/G vs A/A) (p-value = 0.04) and in SNP rs1053049 for the codominant (C/C vs T/T) (p-value = 0.01) and the recessive genetic model (C/C vs T/T-C/T) (p-value = 0.004) all polymorphisms were associated with BMI. In conclusion, it was found that three of the four polymorphisms (rs1053049, rs2267668, rs1801282) selected are associated with the risk of being overweight. Having said that, one has to bear in mind that DNA variants do not fully explain the reasons for being overweight. Therefore more research is needed to make a thorough assessment using the latest genomic methods in sequencing and genotyping, combined with epigenomics, proteomics, transcriptomics, and metabolomics.

Releasing Three Orphaned White Rhinoceroses (Ceratotherium simum) to the Game Reserve in South Africa. Rehabilitation, Translocation and Post-Release Observations.

White rhinoceros (Ceratotherium simum) is one of the most famous victims of poachers in Africa. One of the methods for dealing with decreasing rhino numbers is rehabilitating wounded and/or orphaned animals to successfully release them back into the wild. The status of rescued animal differs among individuals, but general procedures must be established and constantly improved. This study presents the history of successful release of three orphaned white rhino females; rehabilitated for 15 months in Wildlife Rehabilitation Centre in a private game reserve in South Africa. Female A was three years old, female B was one year old and the youngest female was three months old on arrival. The animals were rehabilitated together despite the differences in their age and size, with particular attention paid to keeping them as wild as possible. After being weaned and becoming old enough to go back to the wild, they were released at a distance from the rehabilitation centre, which required immobilization and translocation. Since the rhinos were released, they have been successfully living in the wild. All procedures used in this study proved to be sufficient for preparing the animals for life in the wild and can be recommended for other centres.

Current Trends in Understanding and Managing Equine Rhodococcosis.

The aim of this review was to summarize studies on equine rhodococcosis over the last decade. For many years Rhodococcus equi has remained one of the major health challenges in the equine breeding industry worldwide. Recently, many novel approaches and ideas have been described and some of them were initially implemented into the clinical practice. This study reviews a variety of new information about neonatal susceptibility, clinical appearance, considered and applied diagnostic procedures and treatment alternatives, factors limiting accurate prognosis, ideas regarding environmental management and prophylaxis considerations. Although multiple research were conducted, the main problems such as high morbidity and mortality, a lack of reliable prevention strategies and treatment limitations are still unresolved and require further scientific effort.

Extracellular Vesicles in the Blood of Dogs with Cancer-A Preliminary Study.

Extracellular vesicles (EVs) are a heterogeneous population of submicron-sized structures released during the activation, proliferation, or apoptosis of various types of cells. Due to their size, their role in cell-to-cell communication in cancer is currently being discussed. In blood, the most abundant population of EVs is platelet-derived EVs (PEVs). The aim of this study was to estimate the absolute number and the origin of EVs in the blood of healthy dogs and of dogs with various types of cancer. The EV absolute number and cellular origin were examined by flow cytometry technique. EVs were classified on the basis of surface annexin V expression (phosphatidylserine PS+) and co-expression of specific cellular markers (CD61, CD45, CD3, CD21). The number of PEVs was significantly higher in dogs with cancer (median: 409/µL, range: 42-2748/µL vs. median: 170/µL, range: 101-449/µL in controls). The numbers of EVs derived from leukocytes (control median: 86/µL, range: 40-240/µL; cancer median: 443/µL, range: 44-3 352/µL) and T cells (control median: 5/µL, range: 2-66/µL; cancer median: 108/µL, range: 3-1735/µL) were higher in dogs with neoplasia compared to healthy controls. The estimation of PEV and leukocyte-derived EV counts may provide a useful biological marker in dogs with cancer.