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. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 4, pp. 664–669

Evaluation of the prognostic and predictive value of free light chains in patients with chronic lymphocytic leukemia – preliminary results

Introductionκ and λ serum free light chains (sFLCs) are produced during physiological lymphopoesis by plasmocytes and B lymphocytes in a constant ratio related to heavy chains. The measurement of sFLC plays an important role in the diagnosis and monitoring of patients with multiple myeloma (MM). The first reports suggested that sFLC disturbances might have prognostic value also in patients with chronic lymphocytic leukemia (CLL). Aim of the study: The aim of the study was to evaluate the relationship between sFLC concentration and recognized prognostic factors and clinical course of CLL. Materials and methods: The sFLC concentration was measured using a latex-enhanced immunoassay in 59 patients with newly diagnosed CLL. The relationship between sFLC concentration and time to start of the treatment (TFT), the response rate to therapy (ORR) and overall survival (OS) was assessed. ResultsA significant correlation was found between sFLC κ concentration and the clinical stage of leukemia according to Rai classification, β-2 microglobulin concentration, LDH activity, CD38 expression, as well as between sFLC λlevel and β-2 microglobulin concentration and platelet count (PLT ). There was also a correlation between the values of summated κ and λ and the clinical stage of disease according to Rai classification, β-2 microglobulin concentration, CD38 expression, white blood cells count (WBC), lymphocyte count (ALC) and hemoglobin (Hgb) concentration. The κ/λ ratio (FCLR) values were significantly different in the CD38+ and CD38- population. SummarySimple and reproducible clonality index, which constitutes the sFLC concentration assessment, can be an attractive, potential prognostic marker in patients with CLL, however further studies are needed on a larger group of patients especially in relation to the predictive value of sFLC

Carotid artery volumetric measures associate with clinical ten-year cardiovascular (CV) risk scores and individual traditional CV risk factors in rheumatoid arthritis; a carotid-MRI feasibility study

Background: Common carotid artery intima-media thickness (CIMT), as measured by ultrasound, has utility in stratification of the accelerated cardiovascular risk seen in rheumatoid arthritis (RA); however, the technique has limitations. Carotid magnetic resonance imaging (MRI) is emerging as a useful research tool in the general population, but has yet to be applied in RA populations. Our objectives were to describe the utility of carotid artery MRI (carotid-MRI) in patients with RA in comparison to healthy controls and to describe the association with RA disease phenotype. Methods: Sixty-four patients with RA and no history of cardiovascular (CV) disease/diabetes mellitus were assessed for RA and CV profile, including homeostasis model assessment-estimated insulin resistance (HOMA-IR) and N-terminal pro-brain natriuretic peptide (NT-proBNP). All underwent carotid-MRI (3 T), and were compared to 24 healthy controls. Univariable analysis (UVA) and multivariable linear regression models (MVA) were used to determine associations between disease phenotype and carotid-MRI measures. Results: There were no significant differences in carotid arterial wall measurements between patients with RA and controls. Wall and luminal volume correlated with 10-year CV risk scores (adjusted as per 2017 European League Against Rheumatism (EULAR) guidance); rho = 0.33 (p = 0.012) and rho = 0.35 (p = 0.008), respectively, for Joint British Societies-2 risk score. In UVA, carotid-MRI volumetric measures predominantly were associated with traditional CV risk factors including age, ever-smoking and HOMA-IR (p < 0.05). Lower body mass index was associated with wall maximum thickness (r = − 0.25 p = 0.026). In MVA, age was independently associated with wall volume (B 1.13 (95% CI 0.32, 1.93), p = 0.007) and luminal volume (B 3.69 (95% CI 0.55, 6.83, p = 0.022), and RA disease duration was associated with luminal volume (B 3.88 (95% CI 0.80, 6.97), p = 0.015). Conclusions: This study demonstrates the utility of carotid-MRI in RA, reporting an association between three-dimensional measures in particular and CV risk scores, individual traditional CV risk factors and RA disease duration. Carotid-MRI in RA is a promising research tool in the investigation of CVD

Assessment of bone sialoprotein in the saliva of women at peri- and postmenopausal age

Introduction and objective. Bone sialoprotein (BSP) is present in a non-collagenous fraction of bone matrix, in dentine, cement and calcified cartilage. The characteristic place where sialoprotein is produced makes it highly specific in relation to the bone tissue and the possibility of using it as a marker of bone metabolism. The study aimed to assess the BSP level in the saliva of female patients at the menopausal and postmenopausal period. Materials and method. The study included 71 women aged 45–74 years. In the examined subjects, bone mineral density and bone sialoprotein levels in saliva were determined. Patients also responded to survey questions about place of residence and the frequency of consumption of foods rich in calcium. The obtained results were statistically analyzed. Results. In the study population of women, BSP saliva concentration ranged from 0.40–14.97 ng/ml. The highest BSP values were observed in the control group (average – 5.66 ng/ml, median – 6.73 ng/ml,), the lowest in the osteopenia group (average – 3.09 ng/ml, median – 1.71 ng/ml). This relationship was close to statistical significance (p = 0.056). Place of residence of the surveyed women had neither effect on BSP concentration in saliva nor on the value of the T-score ratio. Similarly, there was no statistically significant relationship between the frequency of consumption of foods rich in calcium and BSP and T-score. Conclusions. It was found that the BSP level can be determined in saliva. Biochemical tests of saliva considering bone metabolism are an issue that should be continued in future, as the availability of material (saliva) for laboratory analyses, carries potential diagnostic opportunities

Assessment of Immune Cell Activation in Pemphigus

(1) Background: Pemphigus is a blistering autoimmune disease of the skin and/or mucous membranes, characterised by the presence of specific autoantibodies directed against structural proteins of the human skin. Recent reports indicate that new haematological parameters, termed Extended Inflammation Parameters (EIP), can be used to assess the activation of immune cells during active inflammation. These include parameters assessing both neutrophil activation (NEUT-RI, NEUT-GI) and the number of activated lymphocytes (RE-LYMP). The aim of this study was to investigate the relationship between changes in NEUT-RI, NEUT-GI and RE-LYMP and the disease activity in patients with pemphigus. (2) Results: The study involved 32 patients with diagnosed different types of pemphigus. Neutrophil activation parameters (NEUT-RI and NEUT-GI) and lymphocytes (RE-LYMP) were significantly higher in these patients compared to the parameters in healthy participants (respectively p = 0.0127, p = 0.0011 and p = 0.0033). The increased quantity of activated lymphocytes (RE-LYMP) also correlated significantly with the extent of skin and/or mucosal lesions in patients assessed by the PDAI scale (p &lt; 0.02). (3) Conclusions: The NEUT-RI, NEUT-GI and RE-LYMP parameters proved to be appropriate markers of inflammation severity in pemphigus, also in relation to local lesions, which was not possible with the inflammation markers (CRP, ESR) used so far on a routine basis

Effect of infliximab on the levels of TNF-alpha and TGF-beta in the whole blood cultures of irradiated patients.

TGF-beta is supposed to be the major cytokine responsible for post-radiation fibrosis of healthy tissues and actively modifies post-radiation changes. The growth of TGF-beta level induces the expression of collagen synthesis gene which triggers off the production of fibrosis of hyaline membranes. The main purpose of this study was to discover the way and methods of reducing post-radiation damage of normal tissues and provide an adequate scientific justification for using Infliximab as an effective radio protector in the neoplasm radiotherapy. A group of 97 patients were subjected to the experiment. Randomly selected patients were assigned to 3 groups according to the radiation exposure. The samples of whole blood were suspended in RPMI 1640 growth medium standardized according to the number of leukocytes. Two milliliters of whole blood was taken from each patient immediately before irradiation and 100 microl sample of the blood was placed in wells with 0.8 mg/ml of Infliximab or without the preparation. TGF-beta levels in blood culture without cA2 before irradiation showed continuous rise from 3978 to 8950 pg/ml at the 96th h. In the post irradiated group without cA2, a continuous growth was recorded till the 48th h (from 4758 to 13324 pg/ml at the 24th h) and then a slight decline to 11950 pg/ml at 96th h, respectively. In the cultures with cA2, TGF-beta levels before irradiation showed also the peak value at the 48th h (from 4050 to 7340 pg/ml at the 48th h) and then started to go down (6500 pg/ml at the 72nd h and 5720 pg/ml at the 96th h). In the post-irradiated group, during the first 6 hours, there was a growth from 4717 pg/ml to 7462 pg/ml, and then a paradoxical increase to 16885 pg/ml at the 12th h. From the 12th h the values started to decrease to 6895 pg/ml at the 96th h. The obtained results confirmed the hypothesis of decreasing the TGF-beta expression by inactivating TNF-alpha with a monoclonal antibody (Infliximab) in the patients' whole blood culture in vitro. These observations are a good starting point for further experiments in vitro and in vivo, whose main objective is to reduce post radiation fibrosis

Effect of infliximab on the levels of TNF-alpha and TGF-beta in the whole blood cultures of irradiated patients.

TGF-beta is supposed to be the major cytokine responsible for post-radiation fibrosis of healthy tissues and actively modifies post-radiation changes. The growth of TGF-beta level induces the expression of collagen synthesis gene which triggers off the production of fibrosis of hyaline membranes. The main purpose of this study was to discover the way and methods of reducing post-radiation damage of normal tissues and provide an adequate scientific justification for using Infliximab as an effective radio protector in the neoplasm radiotherapy. A group of 97 patients were subjected to the experiment. Randomly selected patients were assigned to 3 groups according to the radiation exposure. The samples of whole blood were suspended in RPMI 1640 growth medium standardized according to the number of leukocytes. Two milliliters of whole blood was taken from each patient immediately before irradiation and 100 microl sample of the blood was placed in wells with 0.8 mg/ml of Infliximab or without the preparation. TGF-beta levels in blood culture without cA2 before irradiation showed continuous rise from 3978 to 8950 pg/ml at the 96th h. In the post irradiated group without cA2, a continuous growth was recorded till the 48th h (from 4758 to 13324 pg/ml at the 24th h) and then a slight decline to 11950 pg/ml at 96th h, respectively. In the cultures with cA2, TGF-beta levels before irradiation showed also the peak value at the 48th h (from 4050 to 7340 pg/ml at the 48th h) and then started to go down (6500 pg/ml at the 72nd h and 5720 pg/ml at the 96th h). In the post-irradiated group, during the first 6 hours, there was a growth from 4717 pg/ml to 7462 pg/ml, and then a paradoxical increase to 16885 pg/ml at the 12th h. From the 12th h the values started to decrease to 6895 pg/ml at the 96th h. The obtained results confirmed the hypothesis of decreasing the TGF-beta expression by inactivating TNF-alpha with a monoclonal antibody (Infliximab) in the patients' whole blood culture in vitro. These observations are a good starting point for further experiments in vitro and in vivo, whose main objective is to reduce post radiation fibrosis

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