Conformational studies of alanine-rich peptide using CD and FTIR spectroscopy.

The circular dichroism (CD) and Fourier transform infrared (FTIR) methods were applied to the conformational studies of alanine-rich peptide Ac-K-[A]11-KGGY-NH2 (where K is lysine, A is alanine, G is glycine and Y is tyrozyne) in water, methanol (MeOH) and trifluoroethanol (TFE). The analysis of CD-spectra of the peptide in water at different concentrations revealed that the secondary structure content depends on the peptide concentration and pH of the solution. The increase of the peptide concentration causes a decrease of alpha-helix content and, simultaneously, an increase of beta-sheet structure, while the unordered structure is the predominant one. Additional elements are discovered in MeOH and TFE but alpha-helix and beta-turns predominate. Moreover, in these solutions the percentage content of the secondary structure does not depend on the temperature. FTIR measurements, carried out at higher peptide concentration (about one order of magnitude) than these CD measurements mentioned above, revealed that in water solution the solid state beta-sheet, and aggregated structures, dominate. However, in TFE the most abundant are alpha-helix and beta-turns structures. The thioflavine T assay showed the tendency of the studied peptide for aggregate.     

ABA and gibberellin-like substances during prehardening,cold acclimation,de- and reacclimation of oilseed rape

Previously published results showed that high relative reduction state of PSII (PSII excitation pressure) during both early seedling growth (prehardening) as well as cold deacclimation caused significant changes in growth pattern. The differences in elongation growth rate were related to the cold acclimation of photosynthetic apparatus and to frost resistance. To study changes in the hormonal balance connected with alterations in elongation growth rate observed during prehardening and deacclimation under different PSII excitation pressure (modulated by day-temperatures), endogenous concentration of ABA, GA₃ and GA-like substances (GAs) were analysed. Analyses were also performed during cold acclimation and reacclimation of plants characterized by different elongation growth rate triggered by prehardening or deacclimation under different day-temperatures. Growth under high PSII excitation pressure (prehardening) resulted in a significant increase in ABA and a considerable decrease in GAs contents. On the other hand, different ABA content played almost no role in controlling growth rate during cold deacclimation and subsequent reacclimation, when the induction of elongation growth was connected with the changes in concentration of GAs including GA₃. The possible role of ABA and GAs in controlling prehardening, cold acclimation and deacclimation is discussed.     

In vitro effect of short-term exposure to two synthetic peptides,alone or in combination with clarithromycin or rifabutin,on Cryptosporidium parvum infectivity

The viability of Cryptosporidium parvum after exposure to peptide antibiotics was studied by two different methods, a cell culture system and a double fluorogenic staining. The peptides KFFKFFKFF and IKFLKFLKFL exerted high cytotoxic effects on sporozoites, as demonstrated by cell cultures (complete inhibition after 60 min at 100 microg/ml) and flow cytometry (30% after 20 min at 100 microg/ml), but did not affect consistently the oocysts. Clarithromycin and rifabutin demonstrated less activity against sporozoites but higher activity against oocysts (30% after 180 min at 10 microg/ml). The combination between peptides and azithromycin or rifabutin exerted the highest activities.     

Capillary Zone Electrophoresis method for determination of (+)-S clopidogrel carboxylic acid metabolite in human plasma and urine designed for biopharmaceutic studies

Fast and reproducible Capillary Zone Electrophoresis (CZE) method for the quantification of (+)-S clopidogrel carboxylic acid metabolite in human fluids was elaborated for the first time. Optimal buffer and CZE conditions were established to obtain the complete separation of clopidogrel, its metabolite and piroxicam (internal standard), during one analytical run. Finally, resolution of the analytes was obtained in an uncoated silica capillary filled with a phosphate buffer of pH 2.5. The analytes were isolated from plasma and urine samples using solid phase extraction (SPE). Validation of the CZE method was carried out. The calibration curve of clopidogrel was linear in the range of 0.5–10.0 mg/L in plasma and urine, whereas for (+)-S carboxylic acid metabolite linearity was confirmed in the range of 0.25–20.0 mg/L in plasma and 0.25–10.0 mg/L in urine. Intra- and inter-day precision and accuracy were repeatable. LOD and LOQ were also estimated. SPE recovery of the analytes from plasma and urine was comparable and greater than 80%. The validated method was successfully applied in pharmacokinetic investigations of (+)-S carboxylic acid metabolite of clopidogrel following the oral administration of clopidogrel to patients prior to percutaneous coronary intervention.     

Longidorus poessneckensis Altherr, 1974 and L. piceicola Lišková, Robbins & Brown, 1997 (Nematoda: Longidoridae): new records from Poland and the first description of the L. poessneckensis male and a bivulval female

Longidorus poessneckensis Altherr, 1974 and L. piceicola Lišková, Robbins & Brown, 1997 (Nematoda: Longidoridae) represent new records from Poland. These two species are described and illustrated together with a male and bivulval female of L. poessneckensis. In its general morphology and morphometrics, the male of L. poessneckensis is similar to the females, but has a spicule 100 μm long and one adanal pair, two double and a row of six single ventromedian supplements. Comments on the differential diagnosis of L. poessneckensis and two morphologically related species, L. uroshis Krnjaić, Lamberti, Krnjaić, Agostinelli & Radicci, 2000 and L. macrosoma Hooper, 1961 are given.     

Skeletal and functional status in patients with long-standing stroke

Introduction: Skeletal and functional status can be affected after stroke, mainly due to subjects' immobilisation. The aim of our study was to assess the functional and bone status in post-stroke patients with hemiplegia. Quantitative ultrasound at hand phalanges and bone mineral density (BMD) in the wrist and the calcaneal bone were compared between the paralysed and the non-paralysed side. The study was performed at the Department of Rehabilitation in Nowa Sol. Material and methods: The study included a group of 71 subjects (29 females and 42 males) who had suffered a stroke. Skeletal status was assessed by densitometry (Pixi, Lunar Corp., USA, forearm, calcaneus) and ultrasound (DBM Sonic 1200, IGEA, Italy, hand phalanges). Functional status was evaluated by the Barthel index. Results: The mean values of forearm BMD and ultrasound measurements were significantly lower in the affected limbs versus the opposite site. The mean Barthel index value was 73 ± 21.5. Reduction of the time interval between stroke and standing up positively influenced the difference between wrist BMD in the affected versus the opposite limb, both in the whole group of patients (r = 0.3, p 〈 0.05) and females (r = 0.41, p 〈 0.05). Forearm BMD in males in the affected limb correlated with the Barthel index (r = 0.35, p 〈 0.05). Conclusion: Fracture risk in post-stroke subjects may be increased due to disturbed skeletal and functional status. Reduction of the time interval between stroke and standing up improved wrist densitometric results. (Pol J Endocrinol 2011; 62 (1): 2-7).     

Carbonic anhydrase inhibitors. Selective inhibition of human tumor-associated isozymes IX and XII and cytosolic isozymes I and II with some substituted-2-mercapto-benzenesulfonamides

A series of 2-mercapto-substituted-benzenesulfonamides has been prepared by a unique two-step procedure starting from the corresponding 2-chloro-substituted benzenesulfonamides. Compounds bearing an unsubstituted mercapto group and the corresponding S-benzoyl derivatives were investigated as inhibitors of four isoforms of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), i.e., the cytosolic, ubiquitous isozymes CA I and II, as well as the transmembrane, tumor associated isozymes CA IX and XII. These derivatives were medium potency hCA I inhibitors (K_Is in the range of 1.5–5.7 μM), two derivatives were strong hCA II inhibitors (K_Is in the range of 15–16 nM), whereas the others showed weak activity. These compounds inhibited hCA IX with inhibition constants in the range 160–1950 nM and hCA XII with inhibition constants in the range 1.2–413 nM. Some of these derivatives showed a certain degree of selectivity for inhibition of the tumor-associated over the cytosolic isoforms, being thus interesting leads for the development of potentially novel applications in the management of hypoxic tumors which overexpress CA IX and XII.     

Comparative study of selected blood biochemical components in milk or milk-replacer fed calves during the second week of life

The experiment was carried out on 13 male Polish Black and White dairy calves of 75% share of the Holstein-Friesian (HF) breed during the second week of life. The animals were divided into two groups. One group (n=7) was fed mother's milk and the second (n=6) milk replacer. The dynamics of changes in concentration of selected blood biochemical components connected with nitrogen metabolism (plasma total protein, albumin, urea, endogenous creatinine) and with mineral metabolism (sodium, chloride, calcium, magnesium, zinc, copper and plasma osmotic pressure) were analyzed in both groups. The results show that the type of ingested food influences the concentration of indicators reflecting nitrogen metabolism. Changes of these parameters in calves fed milk replacer are possibly connected with advantageous catabolic changes. Stable concentrations of main extracellular fluid electrolytes and blood plasma osmotic pressure were found in both groups of calves. Constant blood plasma calcium, magnesium, zinc and copper concentrations observed during this study might also indicate the relative maturity of mechanisms maintaining water and electrolyte balance. Nevertheless, it seemsjustifiable to monitor the copper concentrations in plasma of young calves.