Advantageous Extraction, Cleanup, and UHPLC-MS/MS Detection of Patulin Mycotoxin in Dietary Supplements and Herbal Blends Containing Hawberry from Crataegus spp.

Patulin (PAT) is a highly genotoxic mycotoxin still found as the common contaminant of various kinds of spoiled fruits and related commodities which are often endorsed as the health-enhancing products. Thus, a fast and convenient liquid-solid extraction followed by a solid-phase cleanup with the MycoSep®228 AflaPat multifunctional column was used for the highly efficient isolation of PAT with an average recovery of 112.7% from commercial dietary supplements and herbal blends formulated with dried hawberry. Analysis of the PAT content was carried out using gradient elution with a Synergi Polar C18 column (150 × 2 mm, 4 µm) and UHPLC system equipped with a mass spectrometer. PAT was detected in all (n=14) commercial single-component dietary supplements formulated with dried hawberry belonging to Crataegus monogyna and/or Crataegus laevigata. Similarly, PAT was detected in 67% of the studied multicomponent commercial herbal blends (n=6) that contained-in addition to hawberry-different amounts of apple, chokeberry, elderberry, hibiscus, or mallow. Moreover, the PAT content was determined in the hawberry collected from the mature wild hawthorn trees belonging to three botanical species, Crataegus monogyna Jacq., Crataegus laevigata (Poiret) DC, and Crataegus rhipidophylla Gand, growing in the recreational forest areas and in the law-protected state national forest park in Poland. In conclusion, to prevent PAT accumulation and reduce the health risk of consumers in globalizing markets, the implementation of improved cultivation/processing practices of hawthorn trees and hawberry as well as increased analytical control related to the presence of PAT in dietary supplements and herbal blends formulated with fresh, dried, or frozen hawberry should be urgently recommended.

New potential phytotherapeutics obtained from white mulberry (Morus alba L.) leaves.

The present work demonstrates the profound and unique phyto-pharmacological and nutritional profile of white mulberry (Morus alba L.) leaves which containing considerable amounts of easy digestive proteins, carbohydrates, micro- and macronutrients, polyphenols, free amino acids, organic acids. The wide range of significant biopharmaceutical activities of the aqueous and polar organic solvents extracts from mulberry leaves - including antidiabetic, antibacterial, anticancer, cardiovascular, hypolipidemic, antioxidant, antiatherogenic, and anti-inflammatory - have been critically discussed. The main objective was to demonstrate the results of recently published study on the components of white mulberry leaves exhibiting their biological activity in the various pathological and health human ailments. In addition, we intend to drawn the attention of researchers and public health workers for the extended exploration of this deciduous plant leaves as the source of potential indigenous nutraceuticals and functional food products to enable development of alternative prevention and treatment protocols offered in therapy of the common non-communicable diseases and malignances.

Fast Screening of Whole Blood Samples and Pharmaceutical Compounds for Enantiorecognition of Free L-T3 , L-T4 , and D-T4.

A fast screening method of whole blood was proposed for enantiorecognition of free L-T3 , L-T4 , and D-T4 . Stochastic microsensors based on four inulins (IN, IQ, TEX, and HD) immobilized on diamond paste (DP) were used for recognition of free L-T3 , L-T4 , and D-T4 . For the enantiorecognition of free L-T4 and D-T4 in whole blood and pharmaceutical samples, the best microsensor was the one based on TEX/DP (wide linear concentration ranges, and low limits of quantification). The best limit of detection for the assay of free L-T3 (400 fmol/L) was recorded using the microsensors based on HD/DP, while for the assay of free L-T4, and D-T4 the best limit of determination (1 pmol/L) was recorded using the TX/DP-based microsensor. For the enantiorecognition of free L-T3 in whole blood and pharmaceutical samples the best microsensor was the one based on HD/DP (the wider linear concentration range, and the lower limit of quantification - of pmol/L magnitude order). For the enantiorecognition of free L-T3 in whole blood and pharmaceutical samples the best microsensor was the one based on HD/DP (the wider linear concentration range, and the lower limit of quantification - of pmol/L magnitude order). Free L-T3 , L-T4 , and D-T4 were recovered with high reliabilities in whole blood samples (recoveries higher than 99.00%, with RSD values lower than 1.00%) and pharmaceutical samples (recoveries higher than 95.00% with RSD values lower than 1.00%).

A pilot study on the association between double positive Chlamydia pneumoniae serology and serum lipid indices alterations in obese adult females.

AIMS: To study the relationship between Chlamydia pneumoniae serologic status and serum lipid abnormalities in apparently healthy obese female subjects living in urban areas. MAIN METHODS: Serum samples from 117 apparently healthy females (mean age 50 years), classified as overweight/obese (OW/OB,n=74) or normal weight (NW,n=43) according to their body mass index (BMI), were tested for specific IgG and IgA antibodies against C. pneumoniae using ELISA assay. Serum triglycerides (TG), total cholesterol (TChol) and high-density lipoprotein cholesterol (HDLChol) concentrations were measured using enzymatic methods. Serum low-density lipoprotein cholesterol (LDLChol) was calculated using the Friedewald formula. Lifestyle data for all subjects were collected using a validated self-reported questionnaire. KEY FINDINGS: The prevalence of C. pneumoniae infection, as indicated by single positive serology (IgG(+) or IgA(+)), was significantly higher among the OW/OB females than the NW subjects (38.5%vs.27.0%,p=0.021). Females with double-positive serology (IgG(+)/IgA(+)) in the OW/OB group indicated significantly higher (p<0.05) mean serum TG, TChol and LDLChol levels compared to the double seronegatives (IgG(-)/IgA(-)) from this group. On the contrary, no statistical differences (p>0.05) were observed in these serum lipid indices between subjects in the NW group with considered opposite C. pneumoniae serology. Multivariate regression analysis on the 42 double-seropositive subjects, including both OW/OB and NW females and adjusted for such potential confounders as age, BMI and lifestyle factors, showed a significant association of double-positive C. pneumoniae serology with serum TG (ß=0.244;p=0.049),LDLChol (ß=0.332;p=0.037) and TChol-to-HDLChol ratio (ß=0.313;p=0.042). SIGNIFICANCE: These results suggest that possible chronic C. pneumoniae infection is a potential non-dietary factor in modification of the serum lipid profile in the adult OW/OB females.

Proteomic analysis of plasma to reveal the impact of short-term etanercept therapy in pediatric patients with enthesitis-related arthritis: a case report.

In a variety of countries, juvenile idiopathic arthritis (JIA) is the most common cause of chronic arthritis in childhood, yet its etiology is still unknown. In recent years, etanercept, an effective inhibitor of tumor necrosis factor alpha (TNF-alpha), was used as an alternative in certain oligoarticular JIA patients resistant to conventional nonsteroidal anti-inflammatory drugs (NSAIDs), disease-modifying anti-rheumatic drugs (DMARDs), and corticosteroid therapies, and it resulted in sustained improvement in JIA symptoms. This pilot study explores the alterations of specific panels of cytokines and protein profiles in plasma for two Taiwanese pediatric cases with diagnosed enthesitis-related arthritis (ERA), a type of JIA. The patients were studied before and after taking etanercept alone, using a high-content screening approach employing membrane-based human cytokine antibody microarray and the conventional two-dimensional gel electrophoresis (2-DE) proteomic technique. Specifically, 2-DE in combination with mass spectrometry (MALDI-MS) revealed the functional roles of plasma proteins associated with the regulation of immune responses during short-term etanercept treatment of children with ERA. Our study shows that this biotherapy improved clinical ERA manifestations through the regulation of inflammatory mediators, including several cytotoxic cellular cytokines (IL-2/IFN-g), chemokines (MCP-1), and growth factors (GRO) that affect the expression of specific acute phase proteins such as haptoglobins, immunoglobulin A, and fibrinogen-gamma chain. Meanwhile, an up-regulation of antithrombin chain I, vitamin-D binding protein (VDBP), and the various apolipoproteins was also observed after the administration of etanercept in both studied children. These results may be interpreted as the relevant predictive biomarkers of therapeutic responses to etanercept. They suggest that etanercept, which is still rarely used in Taiwan, is a viable treatment for JIA patients, without adverse health effects and increased risk of secondary infections.

Proteomic profiling of oxidative stress in human victims of traffic-related injuries after lower limb revascularization and indication for secondary amputation.

Microsurgical replantation and revascularization are frequently performed to salvage devascularized severe lower-extremity fractures in the human victims of road traffic-related injuries. However, some patients require secondary amputation within 1 week of successful revascularization due to tissue necrosis and sepsis. Enhanced efforts to understand the underlying molecular mechanism of such events are needed and should characterized in depth. Thus, functional proteomics were applied in this study to evaluate the role of oxidative stress in acute injury following microsurgery in a set of human subjects surviving serious road traffic accidents. Changes in the levels of protein volume and the accompanying content in protein carbonylation were visualized using two-dimensional electrophoresis (2-DE) and immunoblot analysis. Since oxidation of some acute-phase proteins not only causes them to lose their function as antioxidants but also initiates the intracellular stress signaling pathway that regulates cytokine and chemokine responses, how cytokine expression correlated with oxidative stress was also evaluated via protein array assays. It was observed that the growth-regulated oncogene protein family (GRO), the range of IL-6, IL-8, IL-10 and monocyte chemotactic protein-1 (MCP-1), which are responsible for neutrophil and monocyte aggregation with subsequent cytotoxic effects, were significantly elevated in the plasma of amputees subjects, whilst the level of chemokine recruiting leucocytes into inflammatory sites (RANTES) was diminished in the salvaged group of patients. Our results suggest that severely oxidative injury during revascularization perturbs the normal redox balance and induces carbonylation of specific proteins, thereby activating pro-inflammatory factors leading to severe tissue damage. The dissimilar 2-DE protein and cytokine profiles revealed here might reflect distinct etiologies resulting in oxidative damage to tissues and may serve as pivotal indicators of local necrosis and the subsequent need for secondary amputation of limbs. We believe that the combination of proteomic and cytokine profile results presented in this work offers more reliable information and defines more sophisticated criteria in clinical practice than currently used C-reactive protein levels (CRP) or white blood cells counts (WBC) for predicting secondary amputation requirements in patients requiring limb salvage surgery.

Proteomic screening of antioxidant effects exhibited by radix Salvia miltiorrhiza aqueous extract in cultured rat aortic smooth muscle cells under homocysteine treatment.

AIM OF THE STUDY: Still little is known about the cellular mechanisms that contribute to the attenuated proliferation of aortic smooth muscle cells under the influence of the oxidative stress factors such as homocysteine (Hcy). Thus, we aimed to evaluate whether Salvia miltiorrhiza Bunge (Labiatae), a Chinese medicinal herb widely used in folk medicine for therapy of variety of human cardiovascular disorders would modulate this Hcy promoted growth effect in model animal aortic cells system. MATERIALS AND METHODS: The Salvia miltiorrhiza roots aqueous extract (SMAE) containing 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenyl lactic acid and salvianolic acid B, as confirmed by narrow-bore HPLC analyses with binary gradient elution was used in variable concentrations for the treatment of the rat aortic smooth muscle A10 cells under Hcy stimulation. Two-dimensional electrophoresis (2-DE) coupled with MALDI-TOF mass spectrometry was applied for the elucidation of protein changes characterizing the response of the rat A10 cells into the Hcy-induced oxidative stress. RESULTS: This study showed that a low dose (0.015 mg/mL) of the SMAE significantly inhibited growth (>60%, p<0.05) of the Hcy stimulated rat A10 cells. In addition, concentration of intracellular reactive oxygen species (ROS) obviously decreased in the rat A10 cells after its incubation with SMAE in terms of catalase increasing activity. Next, marked down-regulation of protein kinase C beta-1 (PKC beta-1) and phosphorylated mitogen-activated protein kinase (p-MAPK) expression suggest that observed inhibitory effect of the polyphenol-rich SMAE on the Hcy-induced growth of rat A10 cells was realized via the PKC/p44/42 MAPK-dependent pathway. The intensity changes of 10 protein spots in response of the rat A10 cells to the Hcy-induced oxidative damage as alpha-4-tropomyosin, vimentin, F1F0-ATP synthase (beta subunit), glucose regulated protein 75 (GRP75), actin (fragment), prohibitin, capping protein, plakoglobin, endoplasmic reticulum protein (ERp29), and peptidylprolyl isomerase A (PPIase A), were detected with statistical significance (p<0.05). Meanwhile, it was showed that used here SMAE resist carbonylation of specific cytoskeleton and chaperone proteins as vimentin, alpha-4-tropomyosin and GRP75, respectively, leading to phenotype transformations in the rat A10 cells. CONCLUSION: These data suggest that applied here SMAE exerts its protective effect through circulating ROS suppression and subsequent modulation of protein carbonylation in rat aortic smooth muscle A10 cells. Redox-proteomics protocol highlighted in this study may be applicable in facilitating the assessing potential novel molecular therapeutic targets to reduce cardiovascular risk related with elevated Hcy levels in various human populations and elucidating new mechanisms through which protein functions can be regulated by the redox status with the use of naturally occurring antioxidants.

Unique potentiometric detection systems for HPLC determination of some steroids in human urine.

Isocratic HPLC with potentiometric detection is used for the determination of some 17-ketosteroids (17-KS), e.g., androsterone, dehydroepiandrosterone and estrone, and their respective sulfated conjugates (17-KSS). Glassy carbon or composite electrodes containing a mixture of graphite and poly(vinyl chloride), PVC, were used as substrate electrodes. These substrates were covered either by montmorillonite or potassium tetrakis(p-chlorophenyl) borate containing PVC-based rubber phase membranes. The neutral 17-KS compounds were derivatized with Girard's reagent P (GP) to obtain cationic pyridinium acetohydrazones prior to the HPLC/potentiometric detection assay. No side reactions were observed, and the GP itself was not interfering. The method yielded accurate and reproducible results and was applicable to samples containing down to micromolar concentrations. Next, the 17-KSS compounds, acting as anionic charged molecules, were determined directly in human urine samples with the HPLC/potentiometry combination without preliminary derivatization. For this purpose, a new anion-sensitive potentiometric electrode was developed using a macrocyclic polyamine containing, PVC-based, rubber phase membrane. The three 17-KSS compounds were also determined accurately down to micromolar concentrations. Especially, the main androgen metabolites as dehydroepiandrosterone sulfate and androsterone sulfate could be selectively determined with a developed potentiometric sensor in human urine samples without time-consuming cleanup and preconcentration step.

An outbreak of body weight dissatisfaction associated with self-perceived BMI and dieting among female pharmacy students.

Some reports indicate that in various groups of society living in the highly developed countries a body weight perception and weight satisfaction tend to be inappropriate when compared with body mass index (BMI) calculated from estimated actual weight and height. Thus in present studies a relationship between body weight perceptions, measured actual BMI, gender, and dieting practices in a sample population of pharmacy students in Poland were examined to verify hypothesis that their incorrect self-perception would provoke occasional, seasonal and permanent eating disorders. Height and weight data of 178 pharmacy students (mean age 22.6+/-2.4 years) in Bydgoszcz, Poland, were collected and validated by completed self-reported questionnaire assessing their self-perceived body weight, desired body weight and past/current dieting practices. Only 34.4% of female and 37.1% of male pharmacy students was satisfied with their current body weight. Statistical analyses revealed significant differences in estimated BMI status (chi(2)=28.5; p=0.0001), desired body weight (chi(2)=15.6; p=0.0004) and past dieting (chi(2)=7.6; p=0.0050) by gender. In the male sub-group of students (n=27) unclear association (chi(2)=6.1; p=0.046) between measured actual BMI status and self-perceived body weight have been presented. Moreover, in male students a significant relationship (chi(2)=4.9; p=0.0261) between actual BMI status and both past as well as current weight control behavior in the form of dieting practices was exhibited. In case of a sub-group of female students (n=151) a diffuse association of actual BMI and self-perception of their body weight (chi(2)=69.5; p=0.0001) was obtained. However, a close relation (chi(2)=16.9; p=0.0007) between actual BMI and only past dieting practices was observed in females. Furthermore, in this last sub-group of students the significant relationship (chi(2)=53.9; p=0.0001) between measured actual BMI and desired body weight was also demonstrated. The study showed an evidence of distorted self-perception of body weight in both sub-groups of considered pharmacy students. There was a tendency to overestimate of body weight in female students, and to underestimate in male students. These results suggest common dissatisfaction of body weight, especially among females, who were more often engaged in dieting, despite not being overweight or obese according to measured actual BMI status.

Hydrophobicity-aided potentiometric detection of catecholamines, beta-agonists, and beta-blockers in a mixed-solvent capillary electrophoresis system.

A series of cationic drug-like substances with distinct basicity, hydrogen-bonding ability, and hydrophobicity, including three catecholamines, two beta-agonists, and thirteen beta-blockers, was successfully detected in a capillary electrophoresis system using an end-capillary coupled potentiometric sensor consisting of a PVC-based liquid membrane deposited directly on a 100 mum diameter copper rod. The electrophoretic separation was performed on a 72 cm x 75 microm id uncoated fused-silica capillary with an acidic background electrolyte containing phosphoric acid in a water-acetonitrile mixture, pH* 2.8. Samples were injected electrokinetically at 5.0 kV for 10 s and a running voltage of 19.5 kV was applied. Excluding the bufuralol/practolol pair, baseline separation of all substances was achieved in the developed CE system within 9 minutes. A linear relationship (R(2) 0.8752) between the sensitivity of the applied potentiometric detector and the parameter log P characterising the hydrophobicity of the analytes was demonstrated. The best observable limits of detection (LODs) were obtained for the highly hydrophobic substances, i. e. bufuralol (8.10 x 10(-8) M injected concentration, S/N = 3), propranolol, alprenolol, and clenbuterol (ca. 1.10 x 10(-7) M). In the case of hydrophilic catecholamines and carbuterol their LODs with potentiometric detection were lowered by a factor of almost one thousand, reaching a value of 6.6 x 10(-5) M.

Supramolecular chiro-biomedical assays and enantioselective HPLC analyses for evaluation of profens as non-steroidal anti-inflammatory drugs, potential anticancer agents and common xenobiotics.

The permanent world-wide increase in therapeutic administration of racemic profens as easy available non-prescribed analgesic drugs and a common first-choice anti-inflammatory agents was recently linked with renewed interest in their beneficial use, also as enantiopure formulations, to treat and/or prevent a variety of human malignancies including its four major types as colorectal, breast, lung, and prostate cancer. This underlies the continuous need of selecting perfectly suited chiral separation methods of profens capable to determine nanolevels of a distomer in presence of the eutomer in a variety of complex biological and environmental media. Thus, current improvements for direct enantiomeric separations of profens by well defined supramolecular-based chiral HPLC and recently developed monolithic, combinatorial, bimodal and polymeric chiral stationary phases employing a modern supramolecular chirality concepts has been outlined in this review. The use of diverse supramolecular approaches for chiral HPLC as an easy accessible tool enabling fast development of nanoscale enantioselective, high-throughput and gradient screening procedures for in situ monitoring of stereoselective ADME properties of profens in range of anticancer drug discovery technologies has been also addressed.

Diversity oriented high-throughput screening of 1,3,4-oxadiazole modified chlorophenylureas and halogenobenzamides by HPLC with peptidomimetic calixarene-bonded stationary phases.

Retention profiles in series of the neutral and highly hydrophobic 1,3,4-oxadiazoles containing chlorophenylurea and halogenobenzamide moiety and indicating analgesic activity were determined in the isocratic standard- and narrow-bore HPLC systems employing, respectively, various octadecylsilica and different calixarene bonded stationary phases. When acetonitrile - 2.65 mM phosphoric acid (55 : 45, %, v/v), pH* 3.25, mobile phase was applied retention of these compounds increased with decline of their overall hydrophobicity according to the general preference of more polar compounds by calixarene cavity in time of its non-specific host-guest supramolecular interactions with halogenated substances. The size of calixarene nanocavity and its upper-rim substitution did not change the observed retention order, resolution and selectivity of separation for oxadiazoles. Compared to the retention on the non-end-capped and the highly-end-capped octadecylsilica HPLC column a most improved separation of some regioisomers of halogenated 1,3,4-oxadiazoles were observed on both used calixarene-type HPLC supports. In addition, preliminary data on the self-assembled supramolecular crystal structure of exemplary 1,3,4-oxadiazolchlorophenylurea with cis-elongated conformation was reported and formation of the monovalent inclusion host-guest complexes between 1,3,4-oxadiazoles and each calixarene-type stationary phase was studied with molecular modelling MM+ and AM1 methods. The structural, isomeric and energetic factors leading to the hydrogen bond stabilized inclusion complexes between these species were considered and used for explanation of observed retention sequence and selectivity of 1,3,4-oxadiazoles separation in applied calixarene-based HPLC systems. All these data would be useful in future development of optimized procedures enabling encapsulation of 1,3,4-oxadiazolurea-type drugs with calixarenes.

Supramolecular systems-based HPLC for chiral separation of beta-adrenergics and beta-adrenolytics in drug discovery schemes.

Increasing amount of data considering polymorphism, splice variants and various affinity states of beta-adrenoceptors has resulted in a new range of opportunities for enantiopure beta-adrenergic and beta-adrenolytic drug discovery and continuous development of reliable high-throughput screening procedures enabling tissue specific pharmacological evaluation of these drugs. Design and fast pharmacological profiling of single enantiomeric molecules combining beta-adrenoceptor affinity with other pharmacophores is also still challenging ability. As the use of chiral stationary phases in HPLC has particularly benefited from results of supramolecular chemistry, this review summarises recent achievements provided by this technique in deciphering of enatiorecognition phenomena affecting pharmacological selectivity of beta-adrenergics and beta-adrenolytics and modifying the efficiency of currently proposed beta-adrenoceptor-targeted therapies. Detailed characteristic of chiral separation performance of these drugs in the range of available supramolecular HPLC systems has also been presented.

Residental factors affecting nutrient intake and nutritional status of female pharmacy students in Bydgoszcz.

The aim of present study was to estimate nutrient intake as well as nutritional status of female pharmacy students from Bydgoszcz, and to investigate relationship of these factors with type of usual residence place during academic year The 24-hour recall method was used to evaluate dietary intake of 47 subjects. Measured values of height, body mass and four skinfolds thickness were used for calculation of BM, FFM, %FM indices. An analysis of nutritional status of studied population showed lower body mass and BMI in the sub-group of female students residing outside of their family home. In comparison to the female students living without parents percentage of energy provided by total fat (29.9%) was significantly less and percentage of energy from carbohydrate was significantly higher (55.4%) than students who reside with their parents. Elevated intake of phosphorus and retinol accompanied by inadequate intake of riboflavin, calcium, iron and copper was exhibited in both residence-type related sub-groups of investigated female pharmacy students.

[Viability of murine 3T3 fibroblasts on the poly(methyl methacrylate) surface modified by constant UV irradiation]. / Przezywalnosc fibroblastów 3T3 na powierzchni poli(metakrylanu metylu) modyfikowanego promieniowaniem UV.

Poly(methyl methacrylate) (PMMA) is commonly applied both in dentistry (bone cement) and in ophthalmology (contact lens, intraocular implants). High adhesion of fibroblasts to bone cement is desirable but keratoprosthesis (intraocular lens--IOL) should be characterized by good transparency and indicate no cells adhesion. Some earlier reports show that sterilization and modification of surface properties of some polymers can be achieved by UV-irradiation which causes a serious physicochemical change in polymer materials to depth of 4 microm. In present studies the surface of PMMA films (30 microm) was continuously irradiated with monochromatic UV-light at 254 nm in varied time intervals. The viability of murine 3T3 fibroblasts cultivated by 12 hours on the surfaces of the non-exposed and the UV-irradiated PMMA in relation to the control fibroblast cell line cultured on the surface of borosilicate glass was estimated with standard fluorescence microscopy procedure. It was found, that 3T3 fibroblasts are highly sensitive to the chemical changes of irradiated surface of PMMA, i.e. increase of surface hydrophilicity and oxidation degree accompanied by decrease of polymer molecular mass and increase of free monomer fraction as determined on the results of contact angle measurements and attenuated total reflection infrared spectra. The percentage of living 3T3 fibroblast cells was 87% after cultivation on the untreated virgine PMMA and only 47% after cultivation on the polymer which was pre-irradiated by 72 h. No further changes in fibroblasts viability (47%) was observed in cultures developed on PMMA after 96 h of its introductory permanent irradiation. However, a period of 48 h of such constant UV-irradiation was quite optimal to obtain a specific modification of PMMA surface leading to significantly increased viability of cultured 3T3 fibroblasts up to the 94%.

[Obesity development associated with viral infections]. / Infekcje wirusowe w etiologii rozwoju otylosci.

This overview of the significance of viral infections in the development of human obesity is presented within context of the commonly recognized obesity risk factors, including the personal and public health consequences of obesity in various countries. In addition, the results of past and recently published studies on the recognition of six taxonomically different viruses which can undoubtedly be associated with obesity progression in some species of animals are summarized. More attention is focused on the results of preliminary epidemiological studies indicating that human infection by the avian adenovirus SMAM-1 or the human adenovirus Ad-36 can be objectively related to symptoms, prevalence, and complications of obesity in some adult men. Proposed pathogenic pathways involved in the observed cases of viral infection-dependent obesity in animals and humans are also briefly described. Urgent implementation of high-throughput diagnostics procedures is advised to extend viral infection-oriented modes of prevention, recognition, and therapy of obesity currently available in modern societies.

[Potentiometric detection of beta-adrenolytic and beta-adrenergic drugs in HPLC systems]. / Potencjometryczna detekcja leków beta-adrenolitycznych oraz beta-adrenergicznych w ukladach HPLC.

This paper presents a comprehensive review of reference literature and the principal results of our own studies on the use of inclusion effects of macrocyclic compounds in the potentiometric detection of hypotensive and antiarrhythmic drugs as well as bronchodilators following their separation in HPLC systems. It was found that the properties of the liquid membrane electrodes embodying some macrocyclic compounds (i.e. trioctylated alpha-cyclodextrin, dibenzo-18-crown-6 or calix[6]arene hexaethylester) enabled reasonable prediction of their sensitivity in terms of the computed hydrophobicity and polarizability parameters of the analyzed drugs. The statistically significant equations enabling the optimalization of the composition of liquid membrane electrodes and the prediction of their detection limits in relation to the molecular structures of cationic drugs were established by applying multiparameter regression procedures. In particular it was indicated that the use of a liquid membrane electrode with trioctylated alpha-cyclodextrin enables sensitive detection (<10(-8) M) of highly hydrophobic drugs (logP >2.5 on the Parham-Hall-Kier scale) such as clenbuterol, bufuralol, and propanolol under HPLC conditions. These results indicate that the developed potentiometric detection method of hypotensive drugs can be considered as a promising alternative to the immunoassay procedures (ELISA) currently recommended for the toxicological determination of trace amounts of the drugs in biological samples. It was also shown that the set of experimentally determined detection limits characterizing the sensitivity of the four different liquid membrane electrodes used in the cation-exchange HPLC system enabled reliable screening and proper pharmacological classification of the beta-adrenergic and beta-adrenolytic drugs analyzed. Using principal component analysis (PCA) of the potentiometric data, it was stated that a differentiation of cardioselective from non-cardioselective beta1-blockers commonly administered as the hypotensive drugs was possible. This result indicates that the new method of potentiometric detection in HPLC systems can be applied as an effective high-throughput screening (HTS) procedure of large combinatorial libraries of newly synthesized drug-like candidates. This may increase the possibilities of identifying the lead compounds to be directed to further specialized biological tests, thus enabling the successful design and formulation of a new generation of more selective and pharmacogenome-oriented hypotensive drugs with reduced side-effects.