Safety and tolerability of therapeutic plasma exchange in autoimmune neurological diseases - a retrospective single-centre analysis.

CLINICAL RATIONALE FOR STUDY: The sudden onset of autoimmune neurological diseases often threatens life. In such clinical situations, fast, effective and safe treatment is needed. Therapeutic plasma exchange (TPE) is an option in the treatment of autoimmune disorders. AIM OF STUDY: The aim was to assess the tolerability and safety of membrane-based therapeutic plasma exchange (mTPE) in patients with autoimmune neurological diseases. MATERIALS AND METHODS: A total of 410 TPE treatments were performed in 91 adult patients. The main reasons for performing TPE were: Guillain-Barre syndrome (39.56%), chronic inflammatory demyelinating polyradiculoneuropathy (20.88%), and myasthenia gravis (17.58%). RESULTS: A total of 183 (44.6%) mTPE treatments were performed without complications. In 18 (19.8%) patients, there were no complications observed in any of the mTPE procedures (a total of 83 procedures). Serious and life-threatening complications occurred during four (0.97%) mTPEs. The most common abnormality in laboratory tests was hypocalcaemia. In patients with a fibrinogen c oncentration ≥ 2 .63 g /L, m easured before the second plasmapheresis, coagulation in the TPE filter was more f requently o bserved (p = 0 .04). CONCLUSIONS AND CLINICAL IMPLICATIONS: Our study proves that the use of plasmapheresis conducted by filtration in the treatment of autoimmune neurological diseases is safe and well tolerated.

Mild hyponatremia discovered within the first 24 hours of ischemic stroke is a risk factor for early post stroke mortality.

BACKGROUND: Comorbidities, complications and laboratory abnormalities are common in stroke patients. One of the common problems is hyponatremia (serum sodium (Na) level <135 mmol/L), but the relationship between hyponatremia and the prognosis in patients with stroke is not well understood. OBJECTIVES: The aim of this study was to investigate the prevalence and severity of hyponatremia, as well as its impact on prognosis in stroke patients on admission to hospital. MATERIAL AND METHODS: The study involved the analysis of the first measurement of the Na level after the admission and its correlations with comorbidities, the scale of clinical assessment of stroke severity (NIHSS), the size and location of the stroke, and mortality. A retrospective study was conducted on 502 patients (among them 263 women) admitted to the hospital on stroke onset (440 ischemic stroke (IS) and 62 hemorrhagic stroke (HS) patients). The post-stroke mortality was defined as early if death occurred within 30 days. RESULTS: Hyponatremia was found in 18.4% of patients with IS and 25.8% of patients with HS, irrespective of age and gender. Hyponatremia is an independent prognostic factor of mortality in people with IS (p = 0.003). Na levels were lower in IS patients who died than in those who remained alive (134.8 ±4.99 mmol/L vs 136.6 ±3.01 mmol/L; p = 0.02). Higher mortality rate was observed among IS patients under 75 years of age and Na level ≤132 mmol/L. In patients with IS, hyponatremia correlates with NIHSS (p = 0.005) and the size and location of the stroke (p = 0.002). CONCLUSIONS: Hyponatremia is more frequently observed in patients with HS than IS. Mild hyponatremia is already known to be an independent prognostic factor in the mortality of people with IS and it may also have value as a prognostic factor in the mortality of the IS population. In a patient with a suspected stroke, there is a need to control electrolyte levels at the onset of the stroke, especially in patients with comorbidities, irrespective of age.

Association of 18bp insertion/deletion polymorphism, at -2549 position of VEGF gene, with diabetic vascular complications in type 2 diabetes mellitus.

PURPOSE: Diabetes mellitus type 2 (T2DM) and its vascular complications are a serious world health problem. For this reason it is important to look for new diabetes complication risk factors. The aim of this study was to determine whether 18-bp insertion/deletion (I/D) polymorphism at -2549 position of the vascular endothelial growth factor (VEGF) gene is associated with diabetic vascular complications (DVC). MATERIAL AND METHODS: Caucasian subjects (n = 100) with T2DM were recruited for this study. Genotyping of the VEGF gene I/D polymorphism was done by the polymerase chain reaction (PCR) method. The results were correlated with laboratory and clinical data. RESULTS: In our population heterozygous of the VEGF gene polymorphism was observed most frequently (57%). DVC were observed in 53 patients. Heterozygous T2DM patients significantly more often suffered from heart failure (HF) and stroke (p = 0.05). Amongst all the DVC, D allele of the VEGF polymorphism had a significantly increased risk of diabetic retinopathy (DR) (OR = 1.31; p = 0.033) irrespective of the duration of diabetes, BMI, the glycemia control expressed by HbA1c, renal function, lipid values or applied treatment. The studied polymorphism did not correlate with coronary heart disease, peripheral vascular disease, cardiovascular death, diabetic kidney disease or applied treatment. CONCLUSIONS: The multivariate logistic regression analysis showed that the D allele in the promoter region of the VEGF gene is an independent risk factor of DR irrespective of other laboratory and clinical variables in T2DM patients. Our study suggests that I/D allele in the studied gene is associated with HF and strokes.

miRNA-146a-5p is upregulated in serum and cartilage samples of patients with osteoarthritis.

INTRODUCTION: Osteoarthritis (OA) is a widely prevalent joint disease leading to motor disability and pain. Appropriate indicators for identifying patients at risk for this progressive disease, identifying molecular events for detecting early phases of the disease, or biomarkers to screen for treatment responses, however, are lacking. Micro RNAs (miRNAs), which play crucial roles in OA, could be potential biomarkers of OA. Because circulating miRNA levels reflect the disease state, they may be useful for OA screening and as diagnostic tools, reducing the need for invasive procedures and minimizing the cost of current diagnostic methods. MATERIALS AND METHODS: The expression levels of 18 microRNAs (let-7e-5p, miR-21-5p, miR-93-5p, miR-101-3p, miR-103a-3p, miR-130a-3p miR-146a-5p, miR-16-5p, miR-193b-3p miR-199a-3p, miR-210-3p, miR-222-3p, miR-22-3p, miR-27a-3p, miR-27b-3p, miR-335-5p, miR-454-3p, and miR-98-5p) were analyzed by quantitative real-time polymerase chain reaction in the cartilage tissues and serum samples of 28 OA patients and were compared to those of 2 healthy controls. RESULTS: Expression of microRNA-146a-5p was significantly upregulated in the cartilage (p=0.006) and serum (p=0.002) of OA patients. The expression levels of miR-146a-5p in the serum were positively correlated with those in the cartilage (Pearson correlation coefficient R=0.32; p=0.002). CONCLUSION: miR-146a-5p was significantly overexpressed in patients with OA, both in the articular cartilage tissue and serum, with a positive correlation between the levels in both types of samples. Therefore, the miR-146a-5p serum level could reflect the molecular processes in the cartilage, suggesting its clinical utility as a biomarker for OA management. Implementing noninvasive biomarker using serum miRNAs involves the analysis of the misregulated miRNAs linked to the cartilage pathology.

MiRNA expression in the cartilage of patients with osteoarthritis.

BACKGROUND: Osteoarthritis (OA), the most prevalent disease of articulating joints, is a complex multifactorial disease caused by genetic, mechanical, and environmental factors. In this research, we evaluated miRNA expression in OA. METHODS: Forty tissue samples from 29 patients undergoing joint replacement for OA were evaluated. Tissue from two control patients undergoing hip replacement not related to OA was used as a control. Total RNA (containing miRNA species) from cartilage was isolated using a miRCURYTM [corrected] Isolation Kit. Expression of 19 miRNAs was assessed by real-time quantitative polymerase chain reaction. RESULTS: Expression of four miRNAs, miR-138-5p, miR-146a-5p, miR-335-5p, and miR-9-5p, was significantly upregulated in OA tissues (patients vs. control group). CONCLUSIONS: These findings may contribute to disease prevention and the development of therapeutic targets for OA.

Different patterns of nuclear and mitochondrial penetration by the G3 PAMAM dendrimer and its biotin-pyridoxal bioconjugate BC-PAMAM in normal and cancer cells in vitro.

The intracellular localization and colocalization of a fluorescently labeled G3 amine-terminated cationic polyamidoamine (PAMAM) dendrimer and its biotin-pyridoxal (BC-PAMAM) bioconjugate were investigated in a concentration-dependent manner in normal human fibroblast (BJ) and squamous epithelial carcinoma (SCC-15) cell lines. After 24 hours treatment, both cell lines revealed different patterns of intracellular dendrimer accumulation depending on their cytotoxic effects. Cancer cells exhibited much higher (20-fold) tolerance for native PAMAM treatment than fibroblasts, whereas BC-PAMAM was significantly toxic only for fibroblasts at 50 µM concentration. Fibroblasts accumulated the native and bioconjugated dendrimers in a concentration-dependent manner at nontoxic range of concentration, with significantly lower bioconjugate loading. After reaching the cytotoxicity level, fluorescein isothiocyanate-PAMAM accumulation remains at high, comparable level. In cancer cells, native PAMAM loading at higher, but not cytotoxic concentrations, was kept at constant level with a sharp increase at toxic concentration. Mander's coefficient calculated for fibroblasts and cancer cells confirmed more efficient native PAMAM penetration as compared to BC-PAMAM. Significant differences in nuclear dendrimer penetration were observed for both cell lines. In cancer cells, PAMAM signals amounted to ~25%-35% of the total nuclei area at all investigated concentrations, with lower level (15%-25%) observed for BC-PAMAM. In fibroblasts, the dendrimer nuclear signal amounted to 15% at nontoxic and up to 70% at toxic concentrations, whereas BC-PAMAM remained at a lower concentration-dependent level (0.3%-20%). Mitochondrial localization of PAMAM and BC-PAMAM revealed similar patterns in both cell lines, depending on the extracellular dendrimer concentration, and presented significantly lower signals from BC-PAMAM, which correlated well with the cytotoxicity.

In vitro cytotoxicity of the ternary PAMAM G3-pyridoxal-biotin bioconjugate.

A third-generation polyamidoamine dendrimer (PAMAM G3) was used as a macromolecular carrier for pyridoxal and biotin. The binary covalent bioconjugate of G3, with nine molecules of biotin per one molecule of G3 (G3(9B)), and the ternary covalent bioconjugate of G3, with nine biotin and ten pyridoxal molecules (G3(9B10P)), were synthesized. The biotin and pyridoxal residues of the bioconjugate were available for carboxylase and transaminase enzymes, as demonstrated in the conversion of pyruvate to oxaloacetate and alanine to pyruvate, respectively, by in vitro monitoring of the reactions, using (1)H nuclear magnetic resonance spectroscopy. The toxicity of the ternary bioconjugate (BC-PAMAM) was studied in vitro on BJ human normal skin fibroblasts and human squamous cell carcinoma (SCC-15) cell cultures in comparison with PAMAM G3, using three cytotoxicity assays (XTT, neutral red, and crystal violet) and an estimation of apoptosis by confocal microscopy detection. The tests have shown that BC-PAMAM has significantly lower cytotoxicity compared with PAMAM. Nonconjugated PAMAM was not cytotoxic at concentrations up to 5 µM (NR) and 10 µM (XTT), and BC-PAMAM was not cytotoxic up to 50 µM (both assays) for both cell lines. It has been also found that normal fibroblasts were more sensitive than SCC to both PAMAM and BC-PAMAM. The effect of PAMAM and BC-PAMAM on the initiation of apoptosis (PAMAM in fibroblasts at 5 µM and BC-PAMAM at 10 µM in both cell lines) corresponded with cytotoxicity assays for both cell lines. We concluded that normal fibroblasts are more sensitive to the cytotoxic effects of the PAMAM G3 dendrimer and that modification of its surface cationic groups by substitution with biologically active molecules significantly decreases that effect, confirming that PAMAM G3 is a useful candidate as a carrier for active biocompound delivery.