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Open access

Irena Kostovska, Tosheska Trajkovska, Sonja Topuzovska, Svetlana Cekovska, Goce Spasovski, Ognen Kostovski and Danica Labudovic

Summary

Background

Diabetic nephropathy (DN) is a leading cause of end-stage renal disease. Progressive damage and decline in the number of podocytes often occur in the early stages of DN. Thus, nephrin as a podocyte-specific protein may be regarded as a potential biomarker of early detection of DN. The aim of this study is to determine whether urinary nephrin is an earlier marker in DN than microalbuminuria and to test the significance of urinary nephrin as a marker for early detection of DN.

Methods

Our cross-sectional study included 90 patients with type 2 diabetes mellitus (T2DM), 30 patients with diagnosed DN and 60 patients without diagnosed DN. As a control group, we used 30 healthy subjects. All patients with T2DM were classified into three subgroups according to urinary microalbumin/creatinine ratio (UMCR): normoalbuminuric, microalbuminuric and macroalbuminuric patients. Nephrin in urine was measured by immunoenzyme assay, microalbumin with turbidimetric and creatinine with the photometric method. In blood sera, we measured a few standard biochemical parameters.

Results

Nephrinuria was found to be present in 100% of patients with T2DM and macroalbuminuria, in 88% with microalbuminuria, as well as 82% of patients with T2DM and normoalbuminuria. A concentration of urinary nephrin was significantly increased in all groups of subjects with T2DM compared to the control group (p<0.05). Nephrinuria correlated statistically negative with eGFR (r=-0.54). ROC analysis showed that nephrin has a total predicted probability of 96% in patients with DN.

Conclusions

Urinary nephrin is earlier, more specific and sensitive marker than microalbumin in early detection of DN.

Open access

Alenka Franko, Katja Goricar, Viljem Kovac, Metoda Dodic-Fikfak and Vita Dolzan

Summary

Background

This study aimed to investigate the association between NLRP3 rs35829419 and CARD8 rs2043211 polymorphisms and the risk of developing pleural plaques, asbestosis, and malignant mesothelioma (MM), and to study the influence of the interactions between polymorphisms and asbestos exposure on the risk of developing these diseases.

Methods

The case-control study included 416 subjects with pleural plaques, 160 patients with asbestosis, 154 subjects with MM and 149 subjects with no asbestos disease. The NLRP3 rs35829419 and CARD8 rs2043211 polymorphisms were determined using real-time PCR-based methods. In the statistical analysis, standard descriptive statistics was followed by univariate and multivariate logistic regression modelling.

Results

Asbestos exposure (medium and high vs low) was associated with the risk for each studied asbestos-related disease. An increased risk of pleural plaques was found for CARD8 rs2043211 AT + TT genotypes (OR = 1.48, 95% CI 1.01–2.16, p = 0.042). When the analysis was performed for MM patients as cases, and pleural plaques patients as controls, a decreased MM risk was observed for carriers of CARD8 rs2043211 TT genotype (OR = 0.52, 95% CI 0.27–1.00, p = 0.049). The interactions between NLRP3 rs35829419 and CARD8 rs2043211 genotypes did not influence the risk of any asbestos-related disease. However, when testing interactions with asbestos exposure, a decreased risk of asbestosis was found for NLRP3 CA+AA genotypes (OR = 0.09, 95% CI 0.01–0.60, p = 0.014).

Conclusions

The results of our study suggest that NLRP3 and CARD8 polymorphisms could affect the risk of asbestos-related diseases.

Open access

Anna Domosławska, Sławomir Zduńczyk and Tomasz Janowski

Abstract

Introduction: Significant improvement of sperm motility within one month effected by oral supplementation of selenium and vitamin E was described in four infertile male dogs which failed to conceive in their last three matings with different bitches.

Material and Methods: The dogs (a Golden Retriever, an English Cocker Spaniel, and two Tibetan Mastiffs) were supplemented daily with selenium (Se) (0.6 mg/kg organic Se yeast) and vitamin E (vit. E) (5 mg/kg) per os for 60 days. Semen was collected on days 0, 30, 60, and 90. The sperm concentration and motility parameters were evaluated by the CASA system, sperm morphology was explored by Diff-Quick staining, and live and dead spermatozoa were differentiated by eosin/nigrosin staining. The concentrations of Se and vit. E were measured in peripheral blood serum on semen collection days.

Results: Before administration, the concentrations of Se in blood plasma were low (86.0–165.0 μg/L). After 30 days of treatment there was an observable improvement in total and progressive sperm motility and kinematic parameters (VAP, VSK, VCL, ALH, BCF, and RAPID). The percentages of live and normal morphology sperm cells were also higher. There was also an observable increase in Se and vitamin E concentrations in blood serum. Bitches were successfully mated and delivered four to six puppies.

Conclusion: Supplementation with Se and vit. E improved rapid sperm motility and restored fertility in infertile dogs with low Se status.

Open access

Maciej Klockiewicz, Małgorzata Sobczak-Filipiak, Tadeusz Jakubowski and Ewa Długosz

Abstract

Introduction: Canine roundworm T. canis and T. leonina infections were investigated in experimentally infected farm mink (Neovison vison) to describe the pattern of pathological lesions in this paratenic host.

Material and Methods: Infections in mink developed following ingestion of embryonated eggs of either parasite or mice tissue infected with both parasite species.

Results: Comparative analysis of haematoxylin- and eosin-stained slides showed essential differences among the experimental groups. The lesions observed included eosinophil and mononuclear inflammatory infiltrates of the intestinal wall and local lymph nodes, inflammation and haemorrhages in liver tissues, and interstitial inflammation and mineralisation of the kidneys and lungs. Larvae migrating through the minks’ bodies also caused particularly salient enlargement of lymphoid follicles in the spleen and inflammatory infiltrates of mononuclear cells in skeletal and heart muscles.

Conclusions: It is assumed that histopathological lesions appeared as a local and general host response to invasive L3 T. canis and T. leonina larvae migrating through the tissues of infected farm mink. Interestingly, mink infected with embryonated eggs had more pronounced lesions than animals infected with tissue larvae. Detailed histopathological examinations of parenchymal organs and striated muscles revealed lesions resembling those observed in other paratenic host species due to toxocarosis.

Open access

Maciej Klockiewicz, Tadeusz Jakubowski, Małgorzata Sobczak-Filipiak, Justyna Bartosik and Ewa Długosz

Abstract

Introduction: Farm mink (Neovison vison) can be naturally exposed to T. canis and T. leonina pathogens on the farm. If mink were hosts, it would imply some veterinary public health as well as animal welfare issues. For this reason, the aim of the study was to determine whether mink might be definitive or paratenic hosts of these parasites. Material and Methods: Four groups of mink were infected with both parasite species using larvated eggs or feed containing mouse tissue previously infected with the parasites. Following inoculation, the infections were monitored in vivo by faecal examination for 14 weeks p.i., and then western blotting and ELISA were performed. Results: Coprology did not reveal any canine roundworm eggs, neither were nematodes found in mink intestines during post mortem examination. The specific IgG antibodies recognising excretory/secretory (ES) antigens of both parasite species were identified in mink sera. Single T. leonina tissue larvae were found in digested organs. Conclusions: Our results confirm that farm mink may contribute both T. canis and T. leonina infections. It was proved that farm mink were not their definitive hosts, and therefore mink faeces need not be considered a source of canine roundworm eggs in any soil it fertilises. Nonetheless, as farm mink may be a paratenic host for both parasite species, this may have some impact on the health and welfare of infected animals.

Open access

Lütfiye Tutkun, Servet Birgin İritaş, Serdar Deniz, Özgür Öztan, Sedat Abuşoğlu, Ali Ünlü, Vugar Ali Türksoy and Sultan Pınar Çetintepe

Summary

Background

Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) are well-known biomarkers of systemic inflammation that have been associated with many diseases in the past. In this study, we aimed to determine the relationship between impaired lung functions and the levels of these biomarkers in DMAc exposed people.

Methods

101 non-exposed control subjects (Group 1) and 109 DMAc-exposed workers from the polyvinyl chloride (PVC) industry were included in the study. In the next step, the exposed group was divided into two groups according to the level of exposure (Group 2 and 3). DMAc, TNF-α, IL-6, creatinine, ALT, AST, GFR and standard spirometry measurements were carried out in all subjects.

Results

When compared to the control group, TNF-α and IL-6 levels were significantly high compatible with the increase of DMAc levels, in the exposed groups. Urinary DMAc Levels were 0.06 mg/L in the control group. This level is significantly low when compared to exposed and severely exposed group (2.43 mg/L and 3.17 mg/L). TNF-α levels were 56.86 pg/mL, 145.52 pg/mL and 230.52 pg/mL in control, exposed and severely exposed groups. IL-6 levels were found to be 38.08 pg/mL, 89.19 pg/mL and 116 pg/mL for control, exposed and severely exposed groups, respectively. Similarly, the FEV1/FVC ratio decreased especially in the severely exposed group (p 0.001).

Conclusions

In our study, results have revealed that TNF-and IL-6 levels are promising biomarkers in the early diagnosis of lung function impairment in inhalational DMAc exposure.

Open access

Dorota Kostrzewa-Nowak, Rafał Buryta and Robert Nowak

Summary

Background

During karate fight muscles work at a very high intensity, and their contractions are extremely strong. The movement pattern contains a great number of feints, dodges, frequent changes in movements’ tempo and direction, hits and kicks, all of which is highly stressful for athlete’s organism, including the immune system.

Methods

T lymphocyte subsets’ distribution and selected cytokines in peripheral blood of three elite karate athletes aged 30 years old (range 21–31 years) with minimum 15 years of training experience were analysed in two experiments: at the beginning of the preparatory phase (a progressive test until exhaustion; an analysis of immune system’s selected parameters and cardiorespiratory fitness measures, including VO2max, VE, AT, MVV, MET, Rf), and during the start-up period (Karate Championships; an analysis of selected parameters of the immune system).

Results

Maximal effort caused an increase in total lymphocyte percentage (p<0.05). A decrease in Th cells in recovery (p<0.05 compared to post-exercise), and an increase in Th naïve cells in recovery (p<0.05) were observed. A significant increase in CD8+ central memory cells (p<0.05) was found only after the progressive test, and no changes in both central and effector memory subsets of CD4+ cells during the first experiment. An increase (p<0.05) in Treg and Th1 and a decrease (p<0.05) in Th2 cells’ distribution during recovery time were found. Additionally, changes (p<0.05) in TNF-α, IL-6, IL-8, IL-10 and IL-12p70 were observed.

Conclusion

Post-effort disorder in immune balance activated compensation pathways involving CD4+ cells. Treg and Th1 cells seem to be subsets of key importance involved in the anabolic effect of physical effort, at least among karate athletes.

Open access

Ana Ninić, Nataša Bogavac-Stanojević, Miron Sopić, Jelena Munjas, Jelena Kotur-Stevuljević, Milica Miljković, Tamara Gojković, Dimitra Kalimanovska-Oštrić and Vesna Spasojević-Kalimanovska

Summary

Background

Coronary artery disease (CAD) is one of the most important causes of mortality and morbidity in wide world population. Dyslipidemia, inflammation and oxidative stress may contribute to disruption of endothelium structure and function, atherosclerosis and CAD. Our study was aimed to determine whether Cu/Zn superoxide dismutase (Cu/Zn SOD) and Mn superoxide dismutase (Mn SOD) gene expression could be modulated by oxidative stress in CAD patients.

Methods

This study included 77 CAD patients and 31 apparently healthy persons. Serum lipid levels, high sensitivity C-reactive protein (hsCRP), total antioxidant status (TAS) and thiobarbituric acid-reacting substances (TBARS) were measured. SOD isoenzymes gene expression was determined in peripheral blood mononuclear cells using quantitative polymerase chain reaction.

Results

Mn SOD messenger ribonucleic acid (mRNA) levels were significantly lower in CAD patients than in controls (p=0.011), while Cu/Zn SOD mRNA levels did not change significantly between tested groups (p=0.091). We found significantly lower high-density lipoprotein-cholesterol (HDL-c) (p<0.001) and TAS (p<0.001) levels and significantly higher hsCRP (p=0.002) and TBARS (p<0.001) in CAD patients than in controls. There were significant positive correlations between TAS and Mn SOD mRNA (ρ=0.243, p=0.020) and TAS and Cu/Zn SOD mRNA (r=0.359, p<0.001). TBARS negatively correlated only with Cu/Zn SOD mRNA (ρ=-0.215, p=0.040). TAS levels remained independent predictor for Mn SOD mRNA levels (OR=2.995, p=0.034).

Conclusions

Results of this study showed that Mn SOD gene expression were decreased in CAD patients compared to controls and can be modulated by non-enzymatic antioxidant status in blood.

Open access

Dragana Jovanović, Marina Roksandić-Milenković, Jelena Kotur-Stevuljević, Vesna Ćeriman, Ivana Vukanić, Natalija Samardžić, Spasoje Popević, Branislav Ilić, Milija Gajić, Marioara Simon, Ioan Simon, Vesna Spasojević-Kalimanovska, Milica Belić, Damjan Mirkov, Zorica Šumarac and Vladislav Milenković

Summary

Background

The objective of this prospective study was to evaluate whether soluble programmed cell death-1/programmed cell death-ligand 1 (PD-1/PD-L1) and serum amyloid A1 (SAA1) are potential diagnostic, predictive or prognostic biomarkers in lung cancer.

Methods

Lung cancer patients (n=115) with advanced metastatic disease, 101 with non-small cell lung cancer, NSCLC (77 EGFR wild-type NSCLC patients on chemotherapy, 15 EGFR mutation positive adenocarcinoma patients, 9 patients with mPD-L1 Expression ≥50% NSCLC – responders to immunotherapy), and 14 patients with small cell lung cancer (SCLC) were examined. ELISA method was used to determine sPD-L1 and SAA1 concentrations in patients’ plasma.

Results

Significantly higher blood concentrations of sPD-L1 and SAA1 were noted in lung cancer patients compared with a healthy control group. In PD-L1+ NSCLC patients, a significantly higher sPD-L1 level was noticed compared to any other lung cancer subgroup, as well as the highest average SAA1 value compared to other subgroups.

Conclusions

It seems that sPD-1/PD-L1 might be a potential biomarker, prognostic and/ or predictive, particularly in patients treated with immunotherapy. Serum amyloid A1 has potential to act as a good predictor of patients’ survival, as well as a biomarker of a more advanced disease, with possibly good capability to predict the course of disease measured at different time points.

Open access

Wen Wang, Ci-You Huang, Zhuo-Ping Wang, Shan-Shan Xu, Tie-Yong Qian, Yi-Ding Chen and Wei-Guo Wu

Summary

Background

The chemokine C-C motif ligand 11, also known as eotaxin-1, has been identified as a novel mediator of inflammatory bone resorption. However, little is known regarding a potential role for CCL11/Eotaxin-1 in postmenopausal osteoporosis.

Objective

The scope of this study was to explore the relationship between serum CCL11/Eotaxin-1 concentrations and disease progression of postmenopausal females with osteoporosis.

Methods

A total of 83 postmenopausal women diagnosed with osteoporosis were enrolled. Meanwhile, 82 postmenopausal women with normal bone mineral density (BMD) and 85 healthy controls inner child-bearing age were enrolled as control. The Dual-energy X-ray absorptiometry was used to examine the BMDs at the femoral neck, lumbar spine 1-4 and total hip of all participants. Serum CCL11/Eotaxin-1 levels were examined by enzyme-linked immunosorbent assay. We also included inflammation marker interleukin-6 (IL-6) as well as a serum marker of bone resorption C-telopeptide cross-linked collagen type 1 (CTX-1). The Visual Analogue Scale (VAS) and Oswestry Disability Index (ODI) were recorded to evaluate the clinical severity in POMP females.

Results

Serum CCL11/Eotaxin-1 levels were significantly elevated in postmenopausal osteoporotic patients PMOP patients compared with PMNOP and healthy controls. We observed a significant negative correlation of serum CCL11/Eotaxin-1 levels with lumbar spine, femoral neck and total hip BMD. Furthermore, serum CCL11/ Eotaxin-1 concentrations were also positively related to the VAS and ODI scores. Last, serum CCL11/ Eotaxin-1 concentrations were positively associated with IL-6 and CTX-1 levels. These correlations remain significant after adjusting for age and BMI. Multivariate linear regression analysis demonstrated that CCL11/Eotaxin-1 could serve as an independent marker.

Conclusions

Serum CCL 11/Eotaxin-1 may serve as a candidate biomarker for postmenopausal osteoporosis. Therapeutics targeting CCL11/Eotaxin-1 and its related signalling way to prevent and slow progression of PMOP deserve further study.