The Impact of the IL-10 Gene Polymorphism on mRNA Expression and IL-10 Serum Concentration in Polish Lupus Patients.

Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by the production of autoantibodies against a lot of nuclear components. Despite many studies on the genetic background of this disease, the pathogenesis remains unclear. The aim of the study is to comprehensively evaluate the polymorphism of the IL-10 promoter gene, its mRNA expression, and the serum IL-10 concentration of SLE female patients and females age-matched controls. Analyzing the association between the level of the tested cytokine and the polymorphism genotype-1082; -819; -592, we found statistically higher serum IL-10 levels in SLE patients compared to in healthy controls (11.9 ± 2.2 pg/mL vs. 9.4 ± 1.7 pg/mL, accordingly; p < 0.0001). We did not find statistically significant differences in the gene polymorphism of IL-10 among SLE patients and controls. The most significant observation derived from our study is that IL-10 mRNA transcripts are upregulated in SLE patients compared to in healthy controls (p < 0.0001). According to our results, the presence of the IL-10 genetic polymorphism has no clinical significance for the development of SLE, and subsequent differences in mRNA and IL-10 concentration results from the influence of other factors which should be the subject of further research.

Could KIM-1 and NGAL levels predict acute kidney injury after paracentesis? - preliminary study.

BACKGROUND: Kidney dysfunction is a common complication in patients with severe liver cirrhosis. There is a need for discovery and validation of novel biomarkers for earlier AKI detection. The aim of this study was to determine if tubular injury markers: NGAL and KIM-1 could be helpful in the early diagnosis of AKI in patients undergoing therapeutic paracentesis. METHODS: This preliminary study included 24 adult patients diagnosed with liver cirrhosis who had been hospitalized due to massive ascites requiring paracentesis. Pre- and post-paracentesis plasma samples were taken from each patient and biomarkers were measured. RESULTS: Before paracentesis, the levels of serum and urinary NGAL were similar between patients and controls; while urinary KIM-1 was markedly increased in liver cirrhotic patients (0.76 vs. 0.24 ng/ml; respectively). Although urinary NGAL levels in AKI patients were 5-time greater than in non-AKI subgroup, the difference did not reach statistical significance (13.2 vs 1.5 pg/mL, p = 0.06). Serum NGAL level, post-procedure, was 3 times greater in AKI subgroup. CONCLUSION: Kidney injury markers, especially serum NGAL, may be useful for the early detection of AKI. However, further research is required to determine if biomarkers of kidney injury may help identify patients with cirrhosis who would most likely benefit from early AKI prevention and treatment.

Mechanism of polypurine tract primer generation by HIV-1 reverse transcriptase.

HIV-1 reverse transcriptase (RT) possesses both DNA polymerase activity and RNase H activity that act in concert to convert single-stranded RNA of the viral genome to double-stranded DNA that is then integrated into the DNA of the infected cell. Reverse transcriptase-catalyzed reverse transcription critically relies on the proper generation of a polypurine tract (PPT) primer. However, the mechanism of PPT primer generation and the features of the PPT sequence that are critical for its recognition by HIV-1 RT remain unclear. Here, we used a chemical cross-linking method together with molecular dynamics simulations and single-molecule assays to study the mechanism of PPT primer generation. We found that the PPT was specifically and properly recognized within covalently tethered HIV-1 RT-nucleic acid complexes. These findings indicated that recognition of the PPT occurs within a stable catalytic complex after its formation. We found that this unique recognition is based on two complementary elements that rely on the PPT sequence: RNase H sequence preference and incompatibility of the poly(rA/dT) tract of the PPT with the nucleic acid conformation that is required for RNase H cleavage. The latter results from rigidity of the poly(rA/dT) tract and leads to base-pair slippage of this sequence upon deformation into a catalytically relevant geometry. In summary, our results reveal an unexpected mechanism of PPT primer generation based on specific dynamic properties of the poly(rA/dT) segment and help advance our understanding of the mechanisms in viral RNA reverse transcription.

Coordination between the polymerase and RNase H activity of HIV-1 reverse transcriptase.

Replication of human immunodeficiency virus 1 (HIV-1) involves conversion of its single-stranded RNA genome to double-stranded DNA, which is integrated into the genome of the host. This conversion is catalyzed by reverse transcriptase (RT), which possesses DNA polymerase and RNase H domains. The available crystal structures suggest that at any given time the RNA/DNA substrate interacts with only one active site of the two domains of HIV-1 RT. Unknown is whether a simultaneous interaction of the substrate with polymerase and RNase H active sites is possible. Therefore, the mechanism of the coordination of the two activities is not fully understood. We performed molecular dynamics simulations to obtain a conformation of the complex in which the unwound RNA/DNA substrate simultaneously interacts with the polymerase and RNase H active sites. When the RNA/DNA hybrid was immobilized at the polymerase active site, RNase H cleavage occurred, experimentally verifying that the substrate can simultaneously interact with both active sites. These findings demonstrate the existence of a transient conformation of the HIV-1 RT substrate complex, which is important for modulating and coordinating the enzymatic activities of HIV-1 RT.

[Listeriosis of central nervous system in patients with ulcerative colitis - case study].

In the recently an increase morbidity inflammatory bowel disease, including ulcerative colitis was observed. The use of purine analogs and their metabolites are associated with a higher incidence of infections in this group of patients. Listeriosis is an infectious disease caused by a relatively anaerobic gram-positive bacteria - Listeria monocytogenes. Common symptoms include fever, nausea, vomiting and diarrhea, but these pathogens can also cause myocarditis, central nervous system infections, including brain abscesses and sepsis. Since the incidence of Listeria monocytogenes is higher in patients with inflammatory bowel disease than in the general population, it is important to pay special attention to this group of patients (in prophylaxis as well as treatment) as these infections are serious and often fatal among them.

[The role of gut microbiota in the pathogenesis of obesity]. / Rola flory jelitowej w patogenezie otylosci.

Obesity is a disease that develops as a result of long-term positive energy balance. In recent years, the influence of gut microflora composition, as a potential factor affecting the energy balance and contributing to fat accumulation, has been studied. It seems that bacteria can affect host energy balance through several mechanisms, such as increased fermentation of undigested polysaccharides and obtaining extra energy from the portion of food, reduced expression of FIAF (fasting-induced adipocyte factor) in the enterocytes with inhibitory activity towards intestinal lipoprotein lipase, and the increased release of peptide YY that slows the intestinal motility. It is also believed that changes in the composition of gut microflora may be one of the factors that induce systemic microinflammation in the obese, an important link in the pathogenesis of obesity related complications, including dyslipidaemia, hypertension and type 2 diabetes. However, the results of previous studies are inconclusive. Many of them have been carried out in an animal model and were not confirmed in studies involving humans. These discrepancies may be due to different composition of the diet, distinct physiological gut microflora and the methodology used in these studies. The present article reviews the current literature on the potential role of gut microflora in the pathogenesis of obesity.

[Vitamin K, bone metabolism and vascular calcification in chronic kidney disease]. / Witamina K a metabolizm kosci i kalcyfikacja naczyn w przewleklej chorobie nerek.

Atherosclerosis is the main cause of morbidity and mortality in the general population, and premature death in patients with chronic kidney disease (CKD) especially dialysis ones. Besides the typical cardiovascular risk factors there is a considerable vascular calcification of intima media in these patients. Vitamin K - dependent proteins play an essential role in the pathogenesis of mineral and bone disorders related to CKD, including vascular calcification. Vitamin K is a family of vitamins, varying in the number of isoprenoid groups (saturated or unsaturated) connected into 2-methyl-1,4-naphthoquinone ring in C3 position. Vitamin K-dependent proteins require carboxylation (VKDPs) for biological activation. The coagulant factors are the most well-known VKDPs, but the role of the other proteins, like Matrix Gla Protein (MGP), Growth Arrest Specific Gene 6 (Gas-6) and osteocalcin has been recently discovered. MGP prevents vascular calcification and Gas-6 affects vascular smooth muscle cell apoptosis and movement. Carboxylation of osteocalcin promotes bone formation. Additionally vitamin K increases proliferation of osteoblasts and apoptosis of osteoclasts, influencing on bone remodeling. There is few studies indicating for decreased consumption of vitamin K in the general population. The restrictive diet recommended for dialysis patients additionally diminishes its daily supply, increasing the chance for vitamin K deficiency in this population. Clinical consequences of inhibition of epoxide reductase by generally used anticoagulants, that inhibiting vitamin K cycle and preventing gamma-carboxylation of Gla proteins, in the peripheral tissue is hardly known. This paper summaries the state of the art knowledge focused on the role of vitamin K in mineral and bone metabolism disorders in CKD patients.

[Diagnostic value of antineutophil cytoplasmic antibodies]. / Znaczenie diagnostyczne przeciwcial przeciw cytoplazmie granulocytów obojetnochlonnych.

Antineutrophil cytoplasmic antibodies (ANCA) constitute a family of auto-antibodies directed against various components of the neutrophil cytoplasm. The indirect immunofluorecence assays detected three fluorescent staining patterns: cANCA--cytoplasmic; pANCA--perinuclear and aANCA--atypical. Occurence ANCA is mainly associated with Wegener's granulomatosis and vasculitis, but they are also detected in autoimmune diseases (eg. in systemic lupus erythematosus, in rheumathoid arthritis, Sjögren's syndrome, in dermatomyositis) and in inflamatory bowel diseases (Crohn disease, colitis ulcerosa). Presence of ANCA was found also in primary sclerosing cholangitis, in chronic infections and in person using some kinds of drugs. The aim of the study was to review recent investigations concerning prevalence of ANCA and their diagnostic value not only for vasculitis but also for the other disease in which they are detected.

The influence of weight loss on anaerobic threshold in obese women.

UNLABELLED: Obesity is associated with decreased physical activity. The aim of the study was to assess the anaerobic threshold in obese and normal weight women and to analyse the effect of weight-reduction therapy on the determined thresholds. PATIENTS AND METHODS: 42 obese women without concomitant disease (age 30.5 ± 6.9y; BMI 33.6 ± 3.7 kg·m(-2)) and 19 healthy normal weight women (age 27.6 ± 7.0y; BMI 21.2 ± 1.9 kg·m(-2)) performed cycle ergometer incremental ramp exercise test up to exhaustion. The test was repeated in 19 obese women after 12.3 ± 4.2% weight loss. The lactate threshold (LT) and the ventilatory threshold (VT) were determined. Obese women had higher lactate (expressed as oxygen consumption) and ventilator threshold than normal weight women. The lactate threshold was higher than ventilatory one both in obese and normal weight women (1.11 ± 0.21 vs 0.88 ± 0.18 L·min(-1), p < 0.001; 0.94 ± 0.15 vs 0.79 ± 0.23 L·min(- 1), p < 0.01, respectively). After weight reduction therapy neither the lactate nor the ventilatory threshold changed significantly. The results concluded that; 1. The higher lactate threshold noted in obese women may be related to the increased fat acid usage in metabolism. 2. Both in obese and normal weight women lactate threshold appears at higher oxygen consumption than ventilatory threshold. 3. The obtained weight reduction, without weight normalisation was insufficient to cause significant changes of lactate and ventilatory thresholds in obese women. Key pointsResults showed that adolescent young female gymnasts have an altered serum inflammatory markers and endothelial activation, compared to their less physically active peers.Physical activities improved immune system.Differences in these biochemical data kept significant after adjustment for body weight and height.

Demyelinating polyneuropathy and lymphoplasmacytic lymphoma coexisting in 36-year-old man: A case report.

BACKGROUND: Lymphoplasmacytic lymphoma is a rare non-Hodgkin's lymphoma, occurring mostly in the elderly. It develops slowly and leads to malignant proliferation of lymphoid line cells in the bone marrow, lymph nodes and spleen. It may also affect nerve roots and meninges; some patients develop sensorimotor polyneuropathy which may precede general symptoms of lymphoma. CASE SUMMARY: We present a case of a 36-year-old man diagnosed in 2012 with chronic inflammatory demyelinating polyneuropathy (CIDP), then he was hospitalized in 2019 due to progressive symptoms of heart failure and significant weight loss over the previous four months. Based on clinical and laboratory findings a diagnosis of lymphoplasmacytic lymphoma was suspected and confirmed by bone marrow flow cytometry. There was no improvement in the results of laboratory tests and the patient's condition after immediate implementation of chemotherapy. Patient died on the fifth day of treatment. CONCLUSION: While CIDP and malignant disease co-occurrence is rare, it should be suspected and investigated in patients with atypical neuropathy symptoms.

Zonulin Family Peptide Levels in Ascites and Serum in Patients with Liver Cirrhosis: A Preliminary Study.

INTRODUCTION: Zonulin is a protein that reversibly modulates the permeability of tight junction of the small intestine wall. As the serum concentration of "zonulin family peptides" (ZFPs) is considered to be a sensitive and useful marker of intestinal wall permeability, its serum level may affect the volume of ascites fluid and change in gut microbiota. The aim of the study was to assess the association between concentrations of ZFPs in serum and ascites in relation to the severity of liver cirrhosis. METHODS: The preliminary study included 24 adult patients diagnosed with alcoholic or viral liver cirrhosis. 18 healthy adult subjects were enrolled as the control group. In patients and controls, there were measured serum and ascites (only in patients) ZFPs, serum bilirubin, creatinine, alanine aminotransferase, total protein, and C-reactive protein (CRP). RESULTS: Cirrhotic patients had lower serum hemoglobin (11.6 vs. 14.3 mg/dL; p < 0.001), platelet count (178 vs. 305 × 103/mm3; p < 0.01), total protein and albumin (58.6 vs. 74.3 g/dL; p < 0.001, 26.6 vs. 42.3 g/dL; p < 0.001, respectively), and serum ZFPs (30.5 vs. 62.0 ng/mL; p < 0.001) in comparison to controls. In patients with cirrhosis serum bilirubin, C-reactive protein level and INR were higher than in controls (3.07 vs. 0.96 mg/dL; 36.9 vs. 5 mg/L; 1.53 vs. 0.95; p < 0.001, respectively). Patients with low ZFP levels were characterized with lower ascites ZFP levels (0.25 vs. 16.4 ng/mL; p < 0.001) and ascites/serum index (0.011 vs. 0.462; p < 0.001). There were negative correlations between ascites ZFPs and platelet count (R = -0.497; p < 0.01) and positive correlation with INR (R = 0.640; p < 0.001). ZFP index positively correlated with platelet count (R = 0.726; p < 0.001) and negatively with INR (R = -0.392; p = 0.06). CONCLUSIONS: Decrease serum ZFP levels seem to reflect their decreased liver synthesis but not increased gut permeability in patients with liver cirrhosis. The physiologically low level of ZFPs in transudate is increased in exudate.

Long-term effects of lipase inhibition by orlistat on gastric emptying and orocecal transit time of a solid meal.

BACKGROUND: We assessed the impact of a prolonged lipase inhibition upon gastric emptying (GE) and orocecal transit time (OCTT) of a 355-kcal low-fat solid meal. METHODS: In double-blind manner, 40 obese women BMI > 30 kg/m2, randomly allocated into two equal groups, took orally t.i.d. 120 mg orlistat or placebo during 8 weeks of a weight-reducing management. At randomization and after 2 months, GE was measured simultaneously with OCTT by means of a 13C-octanoic acid and a hydrogen breath test, respectively. Lipolytic activity was evaluated with a 13C-mixed triglyceride breath test (13C-MTGBT). RESULTS: A profound lipase inhibition by orlistat was confirmed by a 79.5% +/- 16.9% reduction of the cumulative 6-h 13C recovery with 13CMTGBT. GE remained unchanged either in the orlistat (T1/2, 188 +/- 35 min start versus 198 +/- 36 min end) or the placebo (T1/2, 191 +/- 35 min start versus 180 +/- 39 min end) group. OCTT increased from 208 +/- 54 min to 271 +/- 64 min (P < 0.01) after orlistat treatment and did not change significantly (216 +/- 76 vs. 234 +/- 72 min) in the placebo group. CONCLUSIONS: No adverse effect on the GE and a moderate prolongation of the OCTT of a low-fat solid meal is to be expected under a prolonged treatment with orlistat at a typical dosage regimen.

[Obesity and bone metabolism]. / Otylosc a metabolizm kosci.

Both bone and adipose tissue change their size, shape and distribution during the whole human being's life. Many factors, including genetic factors, hormones and activity of nervous system are responsible for these changes. It is generally accepted that obesity has a protective effect on bone tissue. On the other hand some authors present an opposite results--the lack of beneficial effect of obesity on development of osteoporosis fractures. The aim of this article was to present and discuss the relations between adipose tissue and bone metabolism.

Osteoprotegerin--does it play a protective role in the pathogenesis of bone loss in obese perimenopausal women?

INTRODUCTION: Assessment of serum osteoprotegerin (OPG) concentrations in obese patients in comparison to healthy controls and evaluation of a possible correlation between OPG and other markers of bone turnover or calcitropic hormones. MATERIAL AND METHODS: 50 obese perimenopausal women without concomitant diseases (BMI 36.7 +/- 4.1 kg/m(2), mean age 50.4 +/- 4.9 yrs). The control group consisted of 19 healthy women (BMI 24.2 +/- 2.1 kg/m(2); mean age 53.8 +/- 5.1 yrs). In all patients serum concentration of OPG, C telopeptide of type I collagen containing the crosslinking site (CTX), osteocalcin, parathormone (PTH) and vitamin D (25-OH-D(3)) was assessed. Dual energy x-ray absorptiometry (the DXA method) of the lumbar spine and femoral neck was performed using a Lunar DPXL to measure bone marrow density (BMD). RESULTS: In obese perimenopausal women serum OPG, osteocalcin and 25-OH-D(3) levels were significantly lower, and the serum PTH level was significantly higher in comparison to healthy controls. A significantly positive correlation was found between serum OPG level and age in both obese and control subjects. CONCLUSION: The serum OPG level in obese perimenopausal women is significantly lower in comparison to healthy controls and does not correlate significantly with biochemical markers of bone turnover, calcitropic hormones and BMD. It probably cannot play a protective role in the pathogenesis of bone loss in obese perimenopausal women.

Comparative studies of physical properties of kinesiotapes.

BACKGROUND: Nowadays we observe growing popularity of kinesiotaping as a supportive method in physiotherapy. In documents available on kinesiotaping we can find that mechanical properties of tapes are similar to the ones of a human skin, but usually there is hardly any numerical data characterizing these properties. OBJECTIVE: Therefore, testing and comparing physical properties of commercially available kinesiotapes seems to be important. METHODS: Physical properties of five commercially available kinesiotapes were examined. Strain vs. stress data was collected up to 15 N. Program Origin 9.0 was used for data analysis. RESULTS: The obtained results show that up to about 2 N the strain vs. stress characteristics of the tested tapes are similar while for greater stress they differ essentially. CONCLUSIONS: An alternative, to commonly used, way of defining relative strain is proposed. This definition could be more suitable in those cases when desired tape tensions are higher than 50% i.e. in ligament and tendon techniques.

Functional Stability of the Human Kappa Opioid Receptor Reconstituted in Nanodiscs Revealed by a Time-Resolved Scintillation Proximity Assay.

Long-term functional stability of isolated membrane proteins is crucial for many in vitro applications used to elucidate molecular mechanisms, and used for drug screening platforms in modern pharmaceutical industry. Compared to soluble proteins, the understanding at the molecular level of membrane proteins remains a challenge. This is partly due to the difficulty to isolate and simultaneously maintain their structural and functional stability, because of their hydrophobic nature. Here we show, how scintillation proximity assay can be used to analyze time-resolved high-affinity ligand binding to membrane proteins solubilized in various environments. The assay was used to establish conditions that preserved the biological function of isolated human kappa opioid receptor. In detergent solution the receptor lost high-affinity ligand binding to a radiolabelled ligand within minutes at room temperature. After reconstitution in Nanodiscs made of phospholipid bilayer the half-life of high-affinity ligand binding to the majority of receptors increased 70-fold compared to detergent solubilized receptors--a level of stability that is appropriate for further downstream applications. Time-resolved scintillation proximity assay has the potential to screen numerous conditions in parallel to obtain high levels of stable and active membrane proteins, which are intrinsically unstable in detergent solution, and with minimum material consumption.

Functional deficiency of vitamin K in hemodialysis patients in Upper Silesia in Poland.

PURPOSE: Functional vitamin K deficiency (both K1 and K2) is postulated to be one of the most relevant links between chronic kidney disease and vascular calcification in hemodialysis (HD) patients. Recommended dietary restrictions in HD patients superimposed on diversity of eating habits across the countries may affect the prevalence of functional vitamin K deficiency. The aim of this study was to determine the level of functional vitamin K deficiency and its relation to vitamin K1 intake in HD patients in Upper Silesia in Poland. METHODS: Protein-induced vitamin K absence or antagonist-II (PIVKA-II) and undercarboxylated matrix Gla protein (ucMGP) were assessed by ELISA in 153 stable, prevalent HD patients and 20 apparently healthy adults (to establish normal ranges for PIVKA-II and ucMGP). Daily phylloquinone intake was assessed using a food frequency questionnaire. RESULTS: PIVKA-II and ucMGP levels were increased in 27.5 and 77.1 % of HD patients in comparison with the reference ranges in apparently healthy controls, respectively. In 45 % of cases, the increased PIVKA-II level was explained by insufficient phylloquinone intake for Polish population (recommended intake: >55 µg for women and >65 µg for men). Applying ROC analysis, we showed that vitamin K1 intake below 40.2 µg/day was associated with increased PIVKA-II levels. There was no correlation between vitamin K1 intake and plasma concentration of ucMGP, or between PIVKA-II and ucMGP. CONCLUSIONS: (1) Functional vitamin K1 deficiency is explained by low vitamin K1 intake in less than half of HD patients. (2) Undercarboxylated matrix Gla protein level is a poor surrogate for functional vitamin K1 deficiency.

[Impact of the mass-reductive therapy with orlistat on 25-(OH)-D3 and PTH concentration in sera of obese, menopausal women]. / Wplyw terapii odchudzajacej z zastosowaniem Orliastu na stezenie 25-(OH)-D3 i PTH w surowicy u otylych kobiet w wieku okolomenopauzalnym.

UNLABELLED: Epidemiological studies suggest a protective influence of obesity against postmenopausal bone loss. Lower risk of osteoporotic fractures was described in obese patients. However there were only a few studies which examined the effect of weight reduction on bone metabolism and results of these studies are controversial. The aim of the study was to evaluate the influence of weight reduction program using Orlistat on bone metabolism in perimenopausal women. Twenty obese women with simple obesity and without concomitant diseases (BMI 37.1 +/- 3.0 kg/m2, mean age 49.8 +/- 4.6 yrs) were enrolled into this study. The control group consisted of 20 healthy women (mean age 53.5 +/- 5.4 yrs, BMI 24.1 +/- 2.2 kg/m2). All patients have participated in a 3-month weight reduction therapy that consisted of: a 1000-1200 kcal/ day balanced diet (daily calcium consumption about 500mg), Orlistat 3 x 120mg a day and regular physical exercises. Before the weight reduction therapy and after 10% reduction of body weight, serum concentrations of PTH, 25-(OH)-D3, total calcium and phosphorus, total cholesterol were assessed. Dual energy x-ray absorptiometry (DEXA method) of lumbar spine and femoral neck, measuring BMD was performed once, after a 3-month weight reduction therapy using Lunar DPXL. All these measurements were performed only once in control subjects. After a 3-month weight reduction program in patients treated with Orlistat the mean weight loss was 11.6 +/- 5.1 kg which is 12.1 +/- 4.78 %. BMI decreased from 37.1 +/- 3.0 kg/m2 at baseline to 32.6 +/- 2.7 kg/m2 post-treatment. The body weight reduction resulted in significant decrease of body fat and total cholesterol concentration. In obese subjects serum concentration of 25-(OH)-D3 was significantly lower and serum concentration of PTH was significantly higher in comparison to healthy controls, both before and after weight reduction therapy. Serum concentration of PTH, 25-(OH)-D3, total calcium and phosphorus did not change significantly after therapy with Orlistat. CONCLUSION: 3-month weight reduction program using Orlistat did not influence significantly bone metabolism.