Helena Donica

The effect of ursolic and oleanolic acids on human skin fibroblast cells

In this article, we look at how ursolic and oleanolic acids can be used for the purpose of quality control of natural products used in dermatocosmetology as well as of various other therapeutic preparations. Ursolic acid (UA) and oleanolic acid (OA) are pentacyclic triterpenes and they are constituents of many medicinal herbs. In this study, we analyzed the cytotoxic and anti-proliferative activity of OA and UA against normal human skin fibroblasts (HSF). Additionally, the scavenging activity of free radicals of both acids was analyzed. The sensitivity of cells to OA and UA activity was determined using a standard spectrophotometric (MTT) assay. The free radical scavenging activity of OA and UA was measured using the DPPH• test. The F-actin cytoskeletal proteins organization was analyzed using TRITC-phalloidine fluorescent staining. The cytotoxic activity of the analyzed acids was determined using Neutral Red (NR) uptake assay. Of the two isomeric compounds, UA showed a higher cytotoxic activity against HSF cells than did OA. Our investigations showed that OA, in view of its non-toxic nature, may be used as a supplementary factor for dermal preparations.

Comparative analysis of adiponectin isoform distribution in pregnant women with gestational diabetes mellitus and after delivery

To evaluate the distribution of circulating adiponectin isoforms in pregnant women with gestational diabetes mellitus (GDM) and compare isoform distribution changes 12 months after delivery.

Trichophyton mentagrophytes-associated Majocchi's granuloma treated with cryotherapy.

We here report the case of a woman with dermatophytosis of the thighs due to Trichophyton mentagrophytes where an unusual clinical picture posed considerable diagnostic and therapeutic problems. She presented numerous skin lesions located on the dorsolateral face of the left thigh and the medial surface of the right calf. The initial lesions consisted of small itchy pustules that evolved to exfoliation after scratching. Results of histopathologic examination of a skin biopsy were consistent with dermatophytosis, although the negativity of PAS staining did not allow confirmation of this diagnosis. Direct microscopic examination with 10% KOH was negative; however, skin cultures on BioMerieux media revealed Tr. Mentagrophytes. Following the diagnosis of Trichophyton infection, the patient was treated with a combination of isoconazole nitrate and difluocortolone valerate. After therapy, both direct microscopic mycologic examination and culture on BioMerieux medium were negative; however, the lesions persisted, assuming a completely different aspect. Cryotherapy with liquid nitrogen was started. This led to a spectacular improvement: the surface of the skin became almost normal, merely showing slight discoloration. An unusual clinical presentation and non-responsiveness to treatment should prompt revision of the primary diagnosis. A negative result of direct microscopy should not exclude the diagnosis of dermatophytosis. Cryotherapy should be considered in cases that do not respond to conventional antifungal medication.

Significance of plasma chromogranin A determination in neuroendocrine tumour (NET) diagnosis.

The secretory nature of NETs implies the determination of the CgA concentration as a standard marker. The concentration of CgA in plasma correlates with the degree of histopathological differentiation, tumor stage, and is an essential prerequisite for therapy. A retrospective analysis of the results of the plasma CgA concentrations in relation to histopathological and clinical findings (type of NET according to the WHO classification, severity of disease based on the presence of metastases and clinical symptoms) as well as somatostatin receptor scintigraphy was performed in 41 patients with NET. The patients were treated in The Regional Oncology of Lublin from February 2005 to May 2008. Data from the literature and results of this study suggest the use of CgA in the diagnosis and prognosis of NET. Plasma CgA concentration analysed together with histopathological assessment of tumor and the clinical picture is a useful marker in the diagnosis of neuroendocrine tumours. High plasma CgA concentrations may indicate the presence of highly-differentiated NET (WDNEC), and also may indicate the presence of tumor metastasis. The highest CgA concentrations were observed in patients with neuroendocrine tumors associated with carcinoid symptoms and the presence of metastases to the liver.

The New TLC Method for Separation and Determination of Multicomponent Mixtures of Plant Extracts

The new mode of two-dimensional gradient thin layer chromatography (MGD-2D TLC) has been presented. Short distance development of sample in the first dimension leads to formation of the preconcentrated narrow zones. They are consecutively separated in the second dimension with the mobile phase gradient in several steps of development until the eluent reaches the further end of the chromatographic plate. The use of the above-mentioned technique allows isolating and then identifying the compounds of various polarity from the multicomponent mixture. The practical application of two-dimensional gradient thin layer chromatography has been performed for isolation of the two plant (Juniperus and Thymus) oils components as the examples of test mixtures. The experiments have been carried out with the use of silica gel plates as well as a normal phase condition. The results of solute separation with isocratic one-dimensional thin layer chromatography system have been compared with those of two-dimensional gradient system. It has been observed that application of the latter mode leads to almost triplicated number of zones in comparison with the former one. It is purposeful to apply the proposed mode to control the purity of the dominant component or components of the mixture.

FRI0119 Improvement in some, but not all, surrogate measures of cardiovascular disease following intensive treatment of early rheumatoid arthritis

Background Patients with rheumatoid arthritis (RA) have an elevated risk of cardiovascular disease (CVD), predominantly ischaemic heart disease (IHD). Systemic inflammation is thought to be a key contributor to this risk and it is hoped early therapeutic suppression of inflammation will reduce cardiovascular (CV) events in the long term. Insulin resistance, associated with metabolic syndrome, N-terminal pro-brain natriuretic peptide (NT-proBNP) and atherogenic lipid profiles have been proposed as potential surrogate measures of atherosclerosis in RA. Each has been shown to correlate with systemic inflammation, but their utility in RA above measurement of the acute phase response remains unknown. Objectives To determine whether suppression of inflammation in early RA influences surrogate measures of atherosclerosis, including the homeostasis model assessment of insulin resistance (HOMA-IR), NT-proBNP and dyslipidaemia. Methods CRP, fasting glucose and lipids, and in a sub-group, NT-proBNP and fasting insulin, were measured at baseline and week 26 in DMARD naïve RA patients, with 3-12 months symptoms, recruited into the Infliximab as Induction therapy for Early rheumatoid Arthritis (IDEA) multicentre double-blind randomised controlled trial. Patients met the 1987 ACR criteria. Treatment was randomised to IFX +MTX or MTX with 250mg IV methylprednisolone as induction therapy. 120mg IM methylprednisolone was given if DAS>2.4 at 3 time points within the first 26 weeks. HOMA-IR was calculated at each time point. Descriptive statistics, Wilcoxon and paired t-test, and Spearman’s correlation were performed. Results Data from 112 patients (69% female, mean age 53 years, 55% RF and 70% anti-CCP positive) were analysed. At baseline, 9% were on a statin and 14% an anti-hypertensive agent. 4 patients had known IHD and were excluded from the subsequent analysis. The median (range) CRP was 13mg/l (0-30) with CRP normal (<5.0mg/l) in 33%. Baseline CRP negatively correlated with HDL (rho -2.78, p=0.024) and total cholesterol (TC) (rho -2.33, p=0.026). 15%, 30%, 39%, and 25% had at risk levels for HDL, LDL, TC and triglyceride (TG), respectively1. In the subgroup of 79 patients with data on NT-proBNP and fasting insulin, 17% and 38% of patients had raised levels of NT-proBNP and HOMA-IR, respectively. Comparing week 26 to baseline, there was a significant fall in CRP (p<0.001; normal levels in 64%) and HOMA-IR (p=0.008; abnormal in 19%), but not NT-proBNP (p=0.132; elevated in 17%). With respect to the dyslipidaemia, HDL and TC rose (p<0.001 and p<0.001 respectively), but there was no significant change in LDL or TG. 5%, 46%, 61%, and 20% had at risk levels for HDL, LDL, TC and TG, respectively. Conclusions In this study, suppression of inflammation was associated with improvement in insulin resistance and increased HDL, but no significant change in NT-proBNP, TG or LDL following 26 weeks intensive RA treatment. Further analyses of biomarkers at week 78 and drug specific effects are underway. Long term follow up is being performed to determine if those patients with persistently abnormal CV biomarkers develop subclinical or overt CVD in the future despite intensive early treatment for RA. References JAMA. 2001;285(19):2486-97 Disclosure of Interest L.-A. Bissell: None Declared, S. Mackie: None Declared, L. Kozera: None Declared, J. Nam: None Declared, A. Burska: None Declared, E. Hensor: None Declared, H. Keen: None Declared, E. Villeneuve: None Declared, H. Donica: None Declared, P. Conaghan: None Declared, J. Andrews: None Declared, P. Emery: None Declared, A. Morgan Grant/Research support from: Supported in part by a research grant from Investigator-Initiated Studies Program of Merck Sharp & Dohme Limited. The opinions expressed in this paper are those of the authors and do not necessarily represent those of Merck Sharp & Dohme Limited

Serum concentrations of interleukin 18 and 25-hydroxyvitamin D3 correlate with depression severity in men with psoriasis

Objective Psoriasis and depression may have common mechanisms, such as systemic inflammation, dysfunction of the hypothalamic-pituitary-adrenal axis, and vitamin D3 deficiency. Among men with psoriasis, this study examined whether depression severity was associated with serum concentrations of different metabolic and inflammatory markers. Methods The study included 85 men with psoriasis (mean age ± standard deviation [SD], 47 ± 14 years) and 65 men without psoriasis (mean age ± SD, 44 ± 13 years). In both groups, we measured the body mass index; blood pressure; and serum concentrations of lipids, uric acid, lipase, interleukins 6 and 18, cortisol, and 25-hydroxyvitamin D3. All participants completed the Beck Depression Inventory. Other variables analyzed included psoriasis duration, the Psoriasis Area Severity Index, and the percentage of body surface area affected by psoriatic lesions. Results Compared with controls, patients with psoriasis had significantly greater depression severity, higher body mass indices, and higher serum concentrations of total cholesterol and interleukins 6 and 18; moreover, they had significantly lower serum 25-hydroxyvitamin D3 concentrations. In patients with psoriasis, depression severity correlated positively with psoriasis duration, the Psoriasis Area Severity Index, the percentage of body surface area affected by psoriatic lesions, and interleukin-18 concentration. In patients with psoriasis, depression severity correlated negatively with 25-hydroxyvitamin D3 concentration, but it did not correlate significantly with the serum concentrations of interleukin 6 and cortisol. Conclusions High concentrations of interleukin 18 and low concentrations of 25-hydroxyvitamin D3 may be associated with depression severity in men with psoriasis. Thus, further studies should examine whether effective anti-inflammatory treatments or vitamin D3 supplementation can improve depression outcomes in these patients.

The evaluation of 25(OH)D concentration in blood serum of chronic heart failure patients

Abstract This current study examined patients with severe chronic heart failure (CHF) in order to ascertain the held vitamin D, based on an assessment of calcidiol [25(OH)D] concentration. It also identified and evaluated possible correlations between 25(OH)D level and the concentration of total calcium, inorganic phosphates and creatinine concentration in their serum. Herein, venous blood samples were taken from 36 patients with CHF. Diagnosis was confirmed by echocardiographic, as well as by electrocardiographic examinations. In this work, the control group consisted of 41 randomly selected healthy individuals. The results of our study showed that CHF patients had significantly lower concentration of 25(OH)D, as well as total calcium and inorganic phosphates. Moreover, mean creatinine concentration was higher, in comparison to the control group, but did not exhibit statistical significance. As calcium-phosphate homeostasis is regulated by numerous factors, including PTH, neurohormonal factors and calcitriol (1,25(OH)2D), it is possible that vitamin D deficiency may play a significant role in the pathomechanism of CHF, and a lowered 25(OH)D level may be related to progression of the disease.

Serum leptin concentration in patients with type 2 diabetes

Abstract With the increasing importance of early type 2 diabetes (DM2) and obesity detection, it is useful to reevaluate leptin role in these conditions. Our study aimed at investigating circulating leptin concentrations in a group of patients with DM2, and at assessing in detail whether leptin concentrations correlate with selected biochemical, clinical parameters and markers of systemic inflammation in patients with DM2 and in healthy volunteers. In our work, we analysed samples and data drawn from 71 patients aged 61.4 ± 11.7 years, who have been diagnosed with type 2 diabetes, as well as from a healthy control group (HC) consisting of 51 healthy subjects with a mean age of 57.8 ± 13.7 years. Therein, the concentration of leptin in the DM2 patients was significantly higher than in the HC (p < 0.01), with median value of 16.59 (IQR 8.58-33.39) ng/ml in the DM2, vs median value of 6.66 (IQR 4.52-21.40) ng/ml in the HC. In the analysis of variance, higher leptin concentrations were revealed in the DM2 group as compared to the HC, and this figure remained significant after adjusting for gender and age (p < 0.001). Moreover, it was independent of HOMA-IR (p = 0.003). However, the differences in leptin levels between the groups disappeared when additional adjustments for anthropometric parameters (BMI, waist circumference) were applied (p = 0.088). Beyond the aforementioned, significant positive correlations were found in the DM 2 group between leptin level and CRP (r=0.256; p < 0.05) and IL-6 (r = 0.345; p < 0.01). Among the selected variables, only gender and BMI were included in the predictive model explaining the variability of leptin, and, in total, were responsible for 72.6% of the original variation of the studied adipocytokine. The results of this study have led to conclusion that leptin may participate in the complex pathogenesis of DM2 and be a predictor of the development of this disease. As higher concentrations of leptin coexist with obesity, and this situation correlates positively with markers of inflammation (CRP, IL-6), leptin level, hence, should be considered in the pathogenesis of DM2.

Comparison of the free and total light chain assays in serum and urine samples with immunofixation electrophoresis for detecting monoclonal proteins in patients with monoclonal gammopathy

ABSTRACT Monoclonal protein (M-protein) is produced by a malignant clone of plasma cells. Detected in serum and/or urine, this typically indicates multiple myeloma (MM) or other monoclonal gammopathy (MG). In a majority of MM cases, with the production of intact monoclonal immunoglobulin (Ig), malignant plasmocytes and/or B lymphocytes often produce excessive amounts of free light chains (FLCs). Excessive synthesis of FLCs lowers the ability of renal proximal tubules to re-absorb FLCs, which results in abnormally high levels of FLCs in the urine (Bence Jones protein, BJP). In laboratory practice, there are tests available for the quantitative measurement of only FLCs κ and λ or for total light chains (TLCs). These tests measure both free forms and bound in the (Ig) molecules forms as light chains that are evident in the serum and in urine. The purpose of this study was to evaluate the FLCs and TLCs approaches in screening serum and urine samples of patients with MM, doing so in comparison to the results of immunofixation (IFE) assessment. A second purpose was to assess the suitability of the collected material for obtaining the most reliable results. The results of serum FLCs (sFLCs) assays suggest that this approach is of the highest reliability and diagnostic usefulness in the detection of MG with excess production of FLCs, in comparison to other available tests. In our work, when κ band light chains were detected in serum IFE (sIFE), 91% patients had their FLCs concentrations beyond the reference range, whereas 89% patients had increased λ FLCs when λ band light chains were detected in sIFE. We also found abnormal sFLC κ/λ ratios in 86.4% and 88.9% of all subject patients who had κ or λ band light chains detected in their sIFE, respectively.