ВИЗНАЧЕННЯ КЕТОПРОФЕНУ І ЦЕФТІОФУРУ У ВЕТЕРИНАРНИХ ПРЕПАРАТАХ СПЕКТРОФОТОМЕТРИЧНИМ МЕТОДОМ

The aim of the work. Development of a method for determination of ketoprofen and ceftiofur in combined veterinary drugs by spectrophotometric method. Materials and Methods. Veterinary drugs “Procefen-100” and “Kotsefen-200” (LLC ”Vetsyntez”, Kharkiv, Ukraine). The standard samples of ketoprofen, ceftiofur and excipients of the pharmacopoeial purity (Sigma-Aldrich) were used. Spectrophotometric (SF) measurements were performed on a CARY.WIN – UV-VIS-50 (Varian, USA) in quartz cells with 10 mm path length. Results and Discussion. Simultaneously, it is impossible to determine the ceftiofur and ketoprofen in combination drugs by direct spectrophotometry. In the absorption spectra of their model mixtures, there is an overlap of the maxima of detectable substances. The conditions of their separation for the next determination by direct spectrophotometry were experimentally selected by us. The optimal solvent was dichloromethane, in which the ketoprofen dissolves, while ceftiofur remains insoluble. 96 % ethanol was used to dissolve ceftiofur. The presence of excipients does not affect on the spectral characteristics of detected substances, which was demonstrated in the analysis of model mixtures of remedies with and without excipients. The technique consists in the extraction of ketoprofen by dichloromethane and its direct spectrophotometric determination at 254 nm wavelength, as well as a direct spectrophotometric determination at 290 nm wavelength of ceftiofur extracted by ethanol. Conclusions. The conditions of the ketoprofen and ceftiofur separation have been found. On the basis of which the method of separation and their quantitative spectrophotometric determination in two combined veterinary drugs with different ratio of the defined substances was developed. The metrological characteristics of the method were determined, and the results of quantitative determination of ketoprofen and ceftiofur were not statistically different from the results obtained by HPLC (by F- and t-criteria).Мета роботи. Розробка методики визначення кетопрофену та цефтіофуру в комбінованих ветеринарних препаратах спектрофотометричним методом. Матеріали і методи. Ветеринарні препарати «Процефен-100» і «Коцефен-200» (ТОВ «Ветсинтез», Харків, Україна). У роботі використовували стандартні зразки кетопрофену та цефтіофуру і допоміжних речовин фармакопейної чистоти (Sigma-Aldrich). Спектрофотометричні (СФ) вимірювання проводили на скануючому спектрофотометрі CARY.WIN – UV-VIS-50 (Varian, США) у кварцових кюветах із товщиною поглинаючого шару l = 1 см. Результати й обговорення. Одночасно неможливо визначити цефтіофур та кетопрофен у комбінованих лікарських препаратах прямим спектрофото-метруванням. В електронних спектрах поглинання їхніх модельних сумішей спостерігається перекривання максимумів визначуваних речовин. Експериментально підібрані умови їхнього розділення для наступного визначення прямим спектрофотометруванням. Оптимальним розчинником виявився дихлорметан, у якому кетопрофен розчиняється, тоді як цефтіофур залишається нерозчинним. Для розчинення цефтіофуру використовували етанол 96 %. Наявність допоміжних речовин не впливає на спектральні характеристики визначуваних речовин, що було показано при аналізі модельних сумішей препаратів із допоміжними речовинами та без них. Методика полягає у вилученні кетопрофену дихлорметаном і його прямому спектрофотометричному визначенні за довжини хвилі 254 нм, а також прямому спектрофотометричному визначенні цефтіофуру, вилученого етанолом, за довжини хвилі λ = 290 нм. Висновки. Встановлено умови розділення субстанцій кетопрофену та цефтіофуру, на основі чого розроблено методику розділення та кількісного спектрофотометричного визначення у двох комбінованих ветеринарних препаратах із різним співвідношенням визначуваних речовин. Розраховано метрологічні характеристики методики, а отримані результати кількісного визначення кетопрофену і цефтіофуру статистично не відрізняються від результатів отриманих заметодикою ВЕРХ (за F- та t-критеріями)

Screening of chorioamnionitis using volatile organic compound detection in exhaled breath: a pre-clinical proof of concept study

Chorioamnionitis is a major risk factor for preterm birth and an independent risk factor for postnatal morbidity for which currently successful therapies are lacking. Emerging evidence indicates that the timing and duration of intra-amniotic infections are crucial determinants for the stage of developmental injury at birth. Insight into the dynamical changes of organ injury after the onset of chorioamnionitis revealed novel therapeutic windows of opportunity. Importantly, successful development and implementation of therapies in clinical care is currently impeded by a lack of diagnostic tools for early (prenatal) detection and surveillance of intra-amniotic infections. In the current study we questioned whether an intra-amniotic infection could be accurately diagnosed by a specific volatile organic compound (VOC) profile in exhaled breath of pregnant sheep. For this purpose pregnant Texel ewes were inoculated intra-amniotically with Ureaplasma parvum and serial collections of exhaled breath were performed for 6 days. Ureaplasma parvum infection induced a distinct VOC-signature in expired breath of pregnant sheep that was significantly different between day 0 and 1 vs. day 5 and 6. Based on a profile of only 15 discriminatory volatiles, animals could correctly be classified as either infected (day 5 and 6) or not (day 0 and 1) with a sensitivity of 83% and a specificity of 71% and an area under the curve of 0.93. Chemical identification of these distinct VOCs revealed the presence of a lipid peroxidation marker nonanal and various hydrocarbons including n-undecane and n-dodecane. These data indicate that intra-amniotic infections can be detected by VOC analyses of exhaled breath and might provide insight into temporal dynamics of intra-amniotic infection and its underlying pathways. In particular, several of these volatiles are associated with enhanced oxidative stress and undecane and dodecane have been reported as predictive biomarker of spontaneous preterm birth in humans. Applying VOC analysis for the early detection of intra-amniotic infections will lead to appropriate surveillance of these high-risk pregnancies, thereby facilitating appropriate clinical course of action including early treatment of preventative measures for pre-maturity-associated morbidities

Effects of metal-contaminated soils on the accumulation of heavy metals in gotu kola (Centella asiatica) and the potential health risks: a study in Peninsular Malaysia

Centella asiatica is a commonly used medicinal plant in Malaysia. As heavy metal accumulation in medicinal plants which are highly consumed by human is a serious issue, thus the assessment of heavy metals in C. asiatica is important for the safety of consumers. In this study, the heavy metal accumulation in C. asiatica and the potential health risks were investigated. Samples of C. asiatica and surface soils were collected from nine different sites around Peninsular Malaysia. The concentration of six heavy metals namely Cd, Cu, Ni, Fe, Pb and Zn were determined by air-acetylene flame atomic absorption spectrophotometer (AAS). The degree of anthropogenic influence was assessed by calculating the enrichment factor (EF) and index of geoaccumulation (Igeo). The heavy metal uptake into the plant was estimated through the calculation of translocation factor (TF), bioconcentration factor (BCF) and correlation study. Estimated daily intakes (EDI) and target hazard quotients (THQ) were used to determine the potential health risk of consuming C. asiatica. The results showed that the overall surface soil was polluted by Cd, Cu and Pb, while the uptake of Zn and Ni by the plants was high. The value of EDI and THQ showed that the potential of Pb toxicity in C. asiatica was high as well. As heavy metal accumulation was confirmed in C. asiatica, daily consumption of the plant derived from polluted sites in Malaysia was not recommended

Src Kinases Are Required for a Balanced Production of IL-12/IL-23 in Human Dendritic Cells Activated by Toll-Like Receptor Agonists

BACKGROUND: Pathogen recognition by dendritic cells (DC) is crucial for the initiation of both innate and adaptive immune responses. Activation of Toll-like Receptors (TLRs) by microbial molecular patterns leads to the maturation of DC, which present the antigen and activate T cells in secondary lymphoid tissues. Cytokine production by DC is critical for shaping the adaptive immune response by regulating T helper cell differentiation. It was previously shown by our group that Src kinases play a key role in cytokines production during TLR4 activation in human DC. PRINCIPAL FINDINGS: In this work we investigated the role of Src kinases during different TLRs triggering in human monocyte-derived DC (MoDC). We found that Src family kinases are important for a balanced production of inflammatory cytokines by human MoDC upon stimulation of TLR3 and 8 with their respective agonists. Disruption of this equilibrium through pharmacological inhibition of Src kinases alters the DC maturation pattern. In particular, while expression of IL-12 and other inflammatory cytokines depend on Src kinases, the induction of IL-23 and co-stimulatory molecules do not. Accordingly, DC treated with Src inhibitors are not compromised in their ability to induce CD4 T cell proliferation and to promote the Th17 subset survival but are less efficient in inducing Th1 differentiation. CONCLUSIONS: We suggest that the pharmacological modulation of DC maturation has the potential to shape the quality of the adaptive immune response and could be exploited for the treatment of inflammation-related diseases

Fusion of metabolomics and proteomics data for biomarkers discovery: case study on the experimental autoimmune encephalomyelitis

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Determinants of the urinary and serum metabolome in children from six European populations

Background Environment and diet in early life can affect development and health throughout the life course. Metabolic phenotyping of urine and serum represents a complementary systems-wide approach to elucidate environment–health interactions. However, large-scale metabolome studies in children combining analyses of these biological fluids are lacking. Here, we sought to characterise the major determinants of the child metabolome and to define metabolite associations with age, sex, BMI and dietary habits in European children, by exploiting a unique biobank established as part of the Human Early-Life Exposome project (http://www.projecthelix.eu). Methods Metabolic phenotypes of matched urine and serum samples from 1192 children (aged 6–11) recruited from birth cohorts in six European countries were measured using high-throughput 1H nuclear magnetic resonance (NMR) spectroscopy and a targeted LC-MS/MS metabolomic assay (Biocrates AbsoluteIDQ p180 kit). Results We identified both urinary and serum creatinine to be positively associated with age. Metabolic associations to BMI z-score included a novel association with urinary 4-deoxyerythronic acid in addition to valine, serum carnitine, short-chain acylcarnitines (C3, C5), glutamate, BCAAs, lysophosphatidylcholines (lysoPC a C14:0, lysoPC a C16:1, lysoPC a C18:1, lysoPC a C18:2) and sphingolipids (SM C16:0, SM C16:1, SM C18:1). Dietary-metabolite associations included urinary creatine and serum phosphatidylcholines (4) with meat intake, serum phosphatidylcholines (12) with fish, urinary hippurate with vegetables, and urinary proline betaine and hippurate with fruit intake. Population-specific variance (age, sex, BMI, ethnicity, dietary and country of origin) was better captured in the serum than in the urine profile; these factors explained a median of 9.0% variance amongst serum metabolites versus a median of 5.1% amongst urinary metabolites. Metabolic pathway correlations were identified, and concentrations of corresponding metabolites were significantly correlated (r > 0.18) between urine and serum. Conclusions We have established a pan-European reference metabolome for urine and serum of healthy children and gathered critical resources not previously available for future investigations into the influence of the metabolome on child health. The six European cohort populations studied share common metabolic associations with age, sex, BMI z-score and main dietary habits. Furthermore, we have identified a novel metabolic association between threonine catabolism and BMI of children