In Vitro Screening of α-Amylase Inhibition by Selected Terpenes from Essential Oils

Purpose: To assess some terpenes from herbal products for possible inhibitory effects on serum α- amylase in order to ascertain their potential usefulness in the prevention and/or treatment of diabetes Type 2.Methods: Solutions of terpenes (citral, eukalyptol, β-pinene, myrcene, eugenol and terpineol) in deonized water were prepared by ultrasonic and manual mixing in four different concentrations ranging from 0.39 – 5.50 µmol cm-3. Commercial sera (with normal-N and high-H enzyme activity) were used as a source of α-amylase. α-Amylase activity was determined by standard methods using an automated analyzer.Results: All the selected terpenes at their maximal concentrations inhibited α-amylase in N-sera in the range 9.68 – 38.70 and 10.71 - 25.00 % for ultrasonic and manual mixing, respectively, while in H-sera, inhibition was in the range 17.10 - 21.05 and 13.58 – 25.92 % for ultrasonic and manual mixing, respectively. Regardless of the concentration of the inhibitor or the method of mixing, citral was the strongest inhibitor of α-amylase.Conclusion: The selected terpenes, in their appropriate concentrations, influence α-amylase activity to varying degrees. Principal component and agglomerative hierarchical analysis reveal that the most significant factor in α-amylase inhibition is the mode of mixing the samples, rather than their concentrations.Keywords: α-Amylase, β-Pinene, Citral, Diabetes, Essential oil, Eugenol, Eukalyptol, Myrcene, Terpenes, Terpineo

Partial purification and some biochemical properties of neonatal rat cutaneous glutathione S-transferases

1. 1. Previous studies have demonstrated the presence of glutathione S-transferases in the skin of rodents and humans. This study represents the first attempt to purify cytosolic glutathione S-transferases from skin of 3-day-old rats.2. 2. A partial purification of the enzyme was achieved by a two-step procedure: affinity chromatography followed by HPLC. Two peaks, one major (P-1) and one minor (P-2), were resolved by HPLC containing about 82% and 10% of the recovered activity, respectively.3. 3. The major form exhibited an overall purification of about 2270-fold with a specific activity of about 73 [mu]moles/min/mg protein towards 1-chloro-2,4-dinitrobenzene.4. 4. The kinetic data for P-1 yielded mean Km values of 2.39 mM for 1-chloro-2,4-dinitrobenzene and 0.72 mM for reduced glutathione, while the respective average Vmax values were found to be 212 and 101 [mu]moles/min/mg protein.5. 5. Significantly inhibition of enzyme activity was noted in the presence of 0.2 mM HgCl2, 0.63 [mu]M 1.2-naphthoquinone, 1.0 [mu]M triphenyltin chloride, and 12.5 [mu]M 17[beta]-estradiol-3-sulfate.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26995/1/0000562.pd

Parcijalna karakterizacija antibakterijskog peptida koji proizvodi probiotska bakterija Lactobacillus plantarum G2

The aim of this study was the partial characterization of the antimicrobial peptide bacteriocin G, produced by probiotic bacteria Lactobacillus plantarum G(2), which was isolated from a clinical sample of a healthy person. Antimicrobial substance was secreted in the supernatant of an L. plantarum G(2) culture, and showed a diverse spectrum of antimicrobial activity of all the tested strains of the genera Lactobacillus and the pathogenic bacteria Staphylococcus aureus and Salmonella abony. Isoclectric focusing revealed that bacteriocin G(2) is a cationic peptide (pI about 10) with a molecular mass of 2.2 kDa according to tricine sodium dodecyl sulphate polyacrylamide gel electrophoresis, SDS-PAGE. The antimicrobial activity of bactcriocin G(2) was diminished by the proteolytic action of trypsin and proteinase K. Bacteriocin G(2) preserved its biological activity in the temperature range 40-60 degrees C (15 min), which was lost at 80 degrees C. Bacteriocin G(2) was stable in the pH range 2-9, while treatment with 1 % Tween 80 and 1 % urea resulted in increased antimicrobial activity. The probiotic strain L. plantarum G(2) produces the antimicrobial substance proteinaceous in nature with bacteriocin characteristics. Bacteriocin production is one of the key properties of probiotic bacteria with clinical potential as anti-infective agents, which will increase the likelihood of its in vivo efficacy.Cilj ovog istraživanja je bila parcijalna karakterizacija antimikrobnog peptida iz probiotske bakterija Lactobacillus plantarum G2 izolovane iz kliničkog uzorka zdrave osobe. Antimikrobno jedinjenje iz L. plantarum G2, označeno kao bakteriocin G2, dobijeno iz supernatanta bakterijske kulture, pokazalo je širok spektar antimikrobne aktivnosti, inhibirajići rast svih ispitivanh vrsta roda Lactobacillus, kao i patogenih bakterija Staphylococcus aureus i Salmonella abony. Bakteriocin G2 je osetljiv na proteolitičko dejstvo tripsina i proteinaze K. Antimikrobna aktivnost je stabilna u opsegu 40-60°C (15 min), ali se gubi na temperaturi od 80°C. Ustanovljeno je da je bakteriocin stabilan na pH vrednostima između 2 i 9. Dejstvo Tween-a 80 i uree je dovelo do povećane inhibitorne aktivnosti. Prema IEF, bakteriocin G2 je katjonski protein, sa pI vrednošću oko 10, a molekulaska masa određena na osnovu tricin-SDS-PAGE je 2,2 kDa. Probiotski soj L plantarum G2 produkuje antimikrobno jedinjenje proteinske strukture sa karakteristikama bakteriocina. Sinteza bakteriocina je jedna od ključnih osobina probiotskih bakterija koje imaju klinički potencijal kao antiinfektivni agensi, jer značajno povećava verovatnoću njihove in vivo efikasnosti

Difference in symptom severity between early and late grass pollen season in patients with seasonal allergic rhinitis

<p>Abstract</p> <p>Background</p> <p>For the development of forecasts for seasonal allergic rhinitis symptoms, it is essential to understand the relationship between grass pollen concentrations and the symptoms of grass pollen allergic patients.</p> <p>Objective</p> <p>The aim of this study was to delineate this relationship between seasonal allergic rhinitis symptoms and grass pollen concentrations in the Netherlands.</p> <p>Methods</p> <p>Grass pollen allergic patients (n = 80 [2007] - 84 [2008]) were enrolled into the study. They were asked to enter their seasonal allergic rhinitis symptoms (runny nose, sneezing, blocked nose, post nasal drip, and eye symptoms) daily on a scale from 0 to 3 to the study centre either by short message service (SMS) or by internet from May-July 2007 and April-July 2008. Daily pollen counts were used to define the early and the late grass pollen season as the period 'before and during' respectively 'after' the first grass pollen peak (more than 150 pollen/m³).</p> <p>Results</p> <p>At similar grass pollen concentrations, the daily mean of the individual maximum symptom scores reported in the early season were higher as compared to that reported in the late season [differences of -0.41 (2007) and -0.30 (2008)]. This difference could not be explained by medication use by the patients nor by co-sensitization to birch.</p> <p>Conclusions</p> <p>We conclude that seasonal allergic rhinitis symptoms at similar grass pollen concentrations are more severe in the early flowering season as compared to those in the late flowering season. This finding is not only relevant for development of forecasts for seasonal allergic rhinitis symptoms but also for understanding symptom development and planning and analysis of clinical studies.</p

Variations of Plasmid Content in Rickettsia felis

Background: Since its first detection, characterization of R. felis has been a matter of debate, mostly due to the contamination of an initial R. felis culture by R. typhi. However, the first stable culture of R. felis allowed its precise phenotypic and genotypic characterization, and demonstrated that this species belonged to the spotted fever group rickettsiae. Later, its genome sequence revealed the presence of two forms of the same plasmid, physically confirmed by biological data. In a recent article, Gillespie et al. ( PLoS One. 2007; 2( 3): e266.) used a bioinformatic approach to refute the presence of the second plasmid form, and proposed the creation of a specific phylogenetic group for R. felis. Methodology/ Principal Findings: In the present report, we, and five independent international laboratories confirmed unambiguously by PCR the presence of two plasmid forms in R. felis strain URRWXCal(2)(T), but observed that the plasmid content of this species, from none to 2 plasmid forms, may depend on the culture passage history of the studied strain. We also demonstrated that R. felis does not cultivate in Vero cells at 37 degrees C but generates plaques at 30 degrees C. Finally, using a phylogenetic study based on 667 concatenated core genes, we demonstrated the position of R. felis within the spotted fever group. Significance: We demonstrated that R. felis, which unambiguously belongs to the spotted fever group rickettsiae, may contain up to two plasmid forms but this plasmid content is unstable

International Veterinary Epilepsy Task Force consensus proposal: Medical treatment of canine epilepsy in Europe

In Europe, the number of antiepileptic drugs (AEDs) licensed for dogs has grown considerably over the last years. Nevertheless, the same questions remain, which include, 1) when to start treatment, 2) which drug is best used initially, 3) which adjunctive AED can be advised if treatment with the initial drug is unsatisfactory, and 4) when treatment changes should be considered. In this consensus proposal, an overview is given on the aim of AED treatment, when to start long-term treatment in canine epilepsy and which veterinary AEDs are currently in use for dogs. The consensus proposal for drug treatment protocols, 1) is based on current published evidence-based literature, 2) considers the current legal framework of the cascade regulation for the prescription of veterinary drugs in Europe, and 3) reflects the authors’ experience. With this paper it is aimed to provide a consensus for the management of canine idiopathic epilepsy. Furthermore, for the management of structural epilepsy AEDs are inevitable in addition to treating the underlying cause, if possible