Applicability of 2,4-dinitrophenylhydrazine (DNPH) method of protein oxidative damage measurement in the seminal plasma of canine ( Canis lupus familiaris) and stallion ( Equus caballus).

Seminal plasma (SP) proteins are responsible for sperm functional quality. Developing a reliable method to determine the degree of oxidative damage of these proteins is important for establishing semen fertilizing ability. The main aim of the study was to verify the applicability of protein carbonyl derivatives measurement in the SP of canine and stallion, using a method with 2,4-dinitrophenylhydrazine (DNPH). The research material consisted of ejaculates obtained from eight English Springer Spaniels, and from seven half-blood stallions during the breeding and non-breeding season. The content of carbonyl groups in the SP was measured on the basis of the reactions with DNPH. The following reagent variants were used to dissolve protein precipitates: Variant 1 (V1) - 6M Guanidine solution and Variant 2 (V2) - 0.1M NaOH solution. It has been shown that to obtain reliable results for the measurement of protein carbonylated groups in the dog and horse SP, both 6M Guanidine and 0.1M NaOH may be used. A correlation was also found between the number of carbonyl groups and the total protein content in the canine (V1: r = -0.724; V2: r = -0.847) and stallion (V1: r = -0.336; V2: r = -0.334) SP. Additionally, the study showed a higher content (p≤0.05) of protein carbonyl groups in the stallion SP in the non-breeding season compared to the breeding season. The method based on the reaction with DNPH, due to its simplicity and cost effectiveness, appears to be suitable for large-scale application in the determination of the SP proteins oxidative damage in dog and horse semen.

Implementation and Sustainability of European Training Program on Organ Donation in Poland: Results and the Impact on Donation Indicators.

BACKGROUND: Hospital training called ETPOD-Essentials in Organ Donation-was introduced in Poland in 31 hospitals with under-utilized potential of donation. The aim of this study was to assess the effect in hospitals included and not included in program, before and after trainings. METHODS: The number of potential and effective donors, organs used, and number (%) of family refusals were compared at 10 and in 20 months after the training and in equal periods before. RESULTS: In trained hospitals, the number of potential donors increased (17% in 10 months, 10% in 20 months); in remaining hospitals, donors increased in 5% in both periods. In hospitals included in ETPOD, the number of effective donors increased (2% and 4.5%); in the whole country, donors also increased (5.6% and 2.7%). In ETPOD hospitals, the number of utilized organs increased (14.5% and 8.5%); in the rest, the increase was 3% and 7%. In trained hospitals, family refusals increased from 6.9% to 16.2% and from 8.9% to 10.7%; in the whole country, family refusals decreased from 11.7% to 11% in the short term and increased from 9.6% to 12.1% in the long term. CONCLUSIONS: In hospitals involved in the ETPOD program, the increase in organ donation is greater than in the rest of hospitals. Distinct benefit was observed in consent to organ donation.

Serum YKL-40 levels in patients with ovarian cancer and women with BRCA1 gene mutation--comparison to CA 125 antigen.

PURPOSE OF INVESTIGATION: Our work was undertaken to determine the usefulness ofYKL-40 as a tumor marker in patients with ovarian cancer and women with BRCA 1 gene mutations. METHODS: Our study population consisted of 111 patients. They were divided into five study groups: I--newly diagnosed ovarian caner, II--recurrence of ovarian cancer, III--complete remission, IV--benign epithelial tumors and V--patients with BRCA 1 gene mutations. YKL-40 and CA 125 were determined in patient sera. RESULTS: YKL-40 in newly diagnosed ovarian cancer patients was significantly higher (181.17 n/ml) than in patients with BRCA 1 gene mutation (97.74 ng/ml, p < 0.01), women with benign epithelial cancer (57.19 ng/ml, p < 0.005) and patients with ovarian cancer at the time of complete remission (58.12 ng/ml, p < 0.005). Taking 124 ng/ml as a cut-off value for YKL-40 (95th percentile for healthly women) we observed higher levels in 50% of patients from group I and in 38% from group II. CONCLUSIONS: YKL-40 appears to demonstrate no advantage over CA 125 as a biomarker of ovarian cancer, particularly in women with early-stage tumors. More research is needed on carriers of the BRCA 1 gene muation in view of the elevated YKL-40 concentrations in this group.

Determination of quercetin and its glucosides in onion by electrochemical methods.

Quercetin (Q), quercetin-3,4'-diO-beta-glucoside (Q3,4'G), quercetin-3-O-beta-glucoside (Q3G) and quercetin-4'-O-beta-glucoside (Q4'G) were determined in onion bulbs (Allium cepa) by HPLC with amperometric detection after analysis of the hydrodynamic voltammograms of flavonoid standards within the potential range of 50-1000 mV and by cyclic voltammetry (CV) method. The hydrodynamic voltammetric profiles of flavonoids showed that the peak current of Q, Q3G, Q4'G and Q3,4'G increased rapidly when the applied potential exceeded +450 mV (vs. SCE). High sensitivity and low background current were observed at the applied potential of +950 mV (vs. SCE). The lower limits of detection (LOD) were determined at signal-to-noise ratio of 3 and showed the following values: 8.05x10(-8)M (Q), 1.08x10(-7) M (Q3G), 1.22x10(-7) M (Q4'G) and 2.6x10(-7) M (Q3,4'G). The data provided by HPLC-UV-MS confirmed the presence of Q, Q3G, Q4'G and Q3,4'G in 80% methanol extracts of lyophilised onion bulbs. The CV method was applied for the qualitative assessment of onion flavonoids followed by the determination of anodic peak potential (E(a)) of the standards. The qualitative analysis of onion flavonoids was based on the anodic peak current (I(a)) of the extracts after external standards addition. The recorded cyclic voltammograms of the above flavonoid standards showed that all four compounds had well-defined oxidation waves with peak potentials of 310, 390, 482 and 800 mV (vs. Ag/AgCl) for Q, Q3G, Q4'G and Q3,4'G in 50 mM acetate-acetic buffer (pH 5.5) in 80% methanol, respectively. The study proved applicability of the CV method for identifying Q, Q3G, Q4'G and Q3,4'G in onion.

Potentiometric detection of organic acids in liquid chromatography using polymeric liquid membrane electrodes incorporating macrocyclic hexaamines.

Potentiometric detection employing coated-wire electrodes was applied to the determination of organic acids in liquid chromatography (LC). Poly(vinyl chloride)-based liquid membranes, incorporating lipophilic macrocyclic hexaamines as neutral ionophores were used as electrode coatings. The selectivity and sensitivity of the macrocycle-based electrodes were found to be superior to an electrode based on a lipophilic anion exchanger (a quaternary ammonium salt). Sensitive detection was obtained for the di- and tricarboxylic acids tartaric, malonic, malic, citric, fumaric, succinic, pyruvic, 2-oxoglutaric and maleic acids after separation in reversed-phase LC. Detection limits (signal/4sigmanoise=3) of 6 pmol for malonic acid and 2 pmol for maleic acid were attained. The detection was explained using a molecular recognition model. The hexaamine-based potentiometric electrodes had a 1-s response time at 1 ml min(-1) flow-rates. They were stable for at least 4 months, with an intra-electrode variation of 3.2% (n=5).

Contribution of membrane surface charge in the interaction of lead and tin derivatives with model lipid membrane.

The objective of this study was to determine the interaction of modified bilayer lipid membranes (BLMs) with lead and tin organoderivatives. The relative depolarization, of the lipid membrane, caused by organo metals, was used to estimate their activity. Dodecyltrimethylammonium bromide (TMDA) and sodium dodecylsulfonate (AS-12) were used to modify BLMs. The trialkylderivatives of tin (IV) were found to be the most active towards the lipid membrane, whereas dialkyltin (IV) and trialkyllead (IV) derivatives were less active. Also, a correlation exists between depolarization activity and the lipophilicity of organo metal hydrolysis products. The surface charge of modified membranes had a secondary influence on depolarization efficiency of the tin and lead derivatives.

[Hearing loss, disorders of sound localization with the preservation ofsmell and taste in diabetics]. / Uposledzenie sluchu, zaburzenia lokalizacji dzwiekowej oraz zachowanie sie wechu i smaku u chorych na cukrzyce.

Diabetes is often complicated by serious medical conditions which could be related to the development of auditory system and cranial nerves lesions, disorder of sound localisation and decreased olfactory and taste ability. Cranial nerve palsies in diabetes are considered as an integral part of the main disease. Twenty nine children with diabetes and non-diabetics control group, without a history of exposure to noise, ototoxic drugs, or ear disease aged 4-19 years old were examined. Hearing impairment may be present in children with disease duration above 3-5 years and in children with other complications. Sound localisation tests were performed by the method of Zakrzewski from a free auditory field, measuring the angle of directional hearing acuity in 35 diabetics age from 16 to 78 years. Longer duration of diabetes was associated with higher directional hearing acuity. Higher hearing loss was observed with higher directional values in diabetics. The authors investigated smell and taste in 35 diabetics. All patients were treated with insulin. Impairment value of smell identification thresholds was much more frequent than impairment of smell perception. In no case raised threshold of taste perception.

[HPV virus infections and other promotion factors of the carcinogenic process in girls]. / Zakazenie wirusem HPV oraz inne czynniki promocyjne w procesie karcinogenezy u dziewczat.

HPV virus infections and other promotion factors in the development of carcinogenic process at 101 girls from different social groups were evaluated. Chronic infection of the cervix was found at the group of 56 smoking girls, aged 15-18, having sexual contacts with many partners. 10 of them were infected by HPV virus. These data indicate that particular health care and widely understood cervical carcinoma prophylaxis in such groups are necessary.

[Lichen sclerosus in girls--risk group for development of vulvar neoplasms in women]. / Lichen sclerosus u dziewczat--grupa ryzyka rozwoju raka sromu u kobiet?

Lichen sclerosus is one of the vulvar dystrophy, which is rare at children and young women. 20 girls, aged 3 to 14 years, with clinical and histopathological diagnosis--lichen sclerosus without dysplastic lesions--were under the control of The Gynecological Outpatient Clinic of Children and Girls in Szczecin. They were treated with estrogens (estriol with vitamin A--ointment) and vitamin A rich diet. Anti-inflammatory treatment was employed if necessary. After long-term treatment at 8 girls the total recovery was obtained, at 2, clinical symptoms decreased. Other girls are under clinical observation as before. The girls with lichen sclerosus are a risk group of vulvar carcinoma development in mature age.