Cannabidiol Decreases Metalloproteinase Activity and Normalizes Angiogenesis Factor Expression in UVB-Irradiated Keratinocytes from Psoriatic Patients

The development of psoriasis is associated with the consequences of oxidative stress and inflammation leading to metabolic changes locally, in the skin cells, and systemically, in the blood. Therefore, the aim of this study was to analyze the effect of psoriasis vulgaris (PsV) and psoriatic arthritis (PsA) on the basal plasma/keratinocyte levels of matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs), and angiogenesis factors, as well as to evaluate the effect of CBD on these parameters in keratinocytes isolated from psoriatic/healthy individuals with and without in vitro irradiation by UVB. A quantitative chemiluminescent method of detection based on an ELISA protocol and zymography technique was used during analysis. It was shown that activity levels of MMP-9 and TIMP-2 in PsA plasma were higher than in PsV. Changes in the proteolytic activity were accompanied by an increase in markers of angiogenesis (angiopoietin-2, HGF, VEGF, TNFα, PDGF, FGF), where in the specific case of angiopoietin-2 and TNFα, the overexpression in PsV was significantly stronger than in PsA. CBD application to keratinocytes partially restored levels of MMP-1/2/3/7 and TIMP-1/2 (in an effect which was particularly enhanced by UVB irradiation), as well as levels of the examined angiogenic factors except TNFα (levels of which were increased in psoriatic keratinocytes and decreased in healthy keratinocytes). Presented results indicate that CBD may be suggested as an antiangiogenic factor that reduces the proinflammatory action of UVB in psoriatic keratinocytes and partially has a protective effect for healthy keratinocytes.

Evaluation and Comparison with Other High-Sensitivity Methods of Analytical Performance and Measured Values of a New Laboratory Test for Cardiac Troponin I Assay

Bakground The aim of this study was to evaluate both analytical characteristics and clinical results of a new chemiluminescent method for the measurement of cardiac troponin I (cTnI), named VITROS ® High Sensitivity Troponin I Assay, using the VITROS® 3600 automated platform. The results found with this new method were compared to those observed with hs-cTnI ARCHITECT and ECLIA hs-cTnT ELECSYS methods. Methods For evaluation of analytical performance and comparison of clinical results, plasma samples (lithium-heparin), were collected from apparently healthy subjects and patients with cardiovascular diseases. Results The hs-cTnI VITROS method showed values for limit of blank (LoB 0.33 ng/L), limit of detection (LoD, 0.91 ng/L), limit of quantifications at 20% (LoQ 20% CV, 1.82 ng/L), and 10% (LoQ 10% CV, 4,74 ng/L), which are comparable to those previously reported for other hs-cTnI methods. Moreover, the clinical results of the hs-cTnI VITROS method were found to be closely correlated to those of hs-cTnI ARCHITECT (R = 0,9883, N = 198) and ECLIA hs-cTnT Elecsys (R = 0,9704, N = 293) methods. Conclusions The hs-cTnI VITROS method shows analytical performance comparable to other cTnI and cTnT assay. The results of this study confirm that there are significant systematic differences among hs-cTnI methods. Further multicenter studies using larger reference populations are needed in order to obtain a better estimation, especially of the 99° percentile URL values categorized for sex and age of hs-cTnI and hs-cTnT methods.

Antioxidant potential of seminal plasma in normozoospermia and asthenozoospermia

The study objective is to evaluate the antioxidant potential of seminal plasma in normozoospermia and asthenozoospermia.Materials and methods. Samples of ejaculate from 57 men of reproductive age were studied. Standard spermiologic examination and determination of the antioxidant potential of the seminal fluid using an original chemiluminescent method were performed. The method allowed to measure duration of the latent period from the moment of a decrease in luminescence of the solution containing reactive oxygen species (ROS) after addition of the seminal fluid until the most dramatic increase in luminescence corresponding to decreased antioxidant effect of the seminal fluid.Results. In the samples, the latent time varies from 4.6 to 17.5 minutes. Compared with normozoospermia, the antioxidant potential is significantly lower (p = 0.05) in men with asthenozoospermia (about 1.4 times), and even lower in patients with asthenoteratozoospermia (about 1.7 times) (p = 0.03). An inverse weak correlation was found between the activity of leukocytes and latent time in pathospermia (r = –0.23 and –0.18 for asthenozoospermia и asthenoteratozoospermia, respectively). A direct strong correlation was found between ROS-producing sperm activity and latent time in normozoospermia (p = 0.79), and inverse correlation in pathospermia (r = –0.26 and –0.62 for asthenozoospermia и asthenoteratozoospermia, respectively).Conclusion. Pathospermia is characterized by antioxidant seminal plasma deficiency, more pronounced for asthenoteratozoospermia. The antioxidant system of seminal plasma correlates with the ROS-producing ability of leukocytes and, to a greater extent, of the sperm. With normozoospermia, the oxidative balance is maintained, with pathospermia, a higher value of ROS production by sperm corresponds to a lower value of antioxidant capacity.

DETERMINATION OF CYTOKINE STATUS INDICATORS AND ANGIOGENESIS FACTORS IN PATIENTS WITH EXTERNAL GENITAL ENDOMETRIOSIS AND VITAMIN D DEFICIENCY

The study was performed to find an association between some cytokine status indicators, level of vascular endothelial growth factors, and vitamin D in women with infertility and external genital endometriosis (EGE) in order to increase the effectiveness of treatment of this pathology. The low vitamin D status in the dynamics was determined in 240 patients aged 25 to 35 years with EGE planning pregnancy by determining the level of 25 (OH) D in the blood serum using the chemiluminescent method. Interleukin status (IL-1β, IL-6, IL-4), TNF-α, VEGF-R1 in blood serum were determined using enzyme-multiple immunoassay. The results of the studies revealed an increased level of IL-6, IL-1β, and TNF-α in groups with low vitamin D content. In normal vitamin D significantly lower levels of vascular endothelial growth factor (VEGF-R1) in the blood serum were registered. Pregnancy rates were higher in groups with normal 25 (OH) D levels in the blood serum. The mean serum VEGF-R1 level in pregnant women who became pregnant on their own was 1.3-1.5 times lower.

A Portable Chemiluminescence Assay of Alkaline Phosphatase Activity to Monitor Pasteurization of Milk Products

ABSTRACT A chemiluminescence assay using a handheld luminometer to measure the activity of alkaline phosphatase was developed that can detect 0.002% or more of unpasteurized milk in various milk products. Evaluation of the assay followed an National Conference on Interstate Milk Shipments (NCIMS)–approved protocol in which aliquots of pasteurized milk products were spiked with raw milk at various levels. Milk products evaluated included skim white milk, 1 and 2% fat content white milk, whole white milk, strawberry-flavored 1% fat content milk, chocolate-flavored 1% fat content milk, half-and-half, and heavy cream. Split samples were prepared, and alkaline phosphatase activities were determined in triplicate on 4 days by three NCIMS-accredited laboratories by the chemiluminescent method and NCIMS-approved reference methods. Equivalence of the chemiluminescent method to the approved reference methods was demonstrated for all eight products evaluated over a range of raw milk concentration from 0 to 0.5%, using criteria established by NCIMS, in which mean results obtained by the three laboratories by the chemiluminescent method were within 1 standard deviation of the mean results obtained by the NCIMS-approved reference methods at each alkaline phosphatase concentration. Procedures for measurement of microbial and reactivated alkaline phosphatase were also established for the method. HIGHLIGHTS