The A/A genotype of an interleukin-17A polymorphism predisposes to increased severity of atopic dermatitis and coexistence with asthma.

INTRODUCTION: Studies have found that the interleukin-23/T helper 17 (IL-23/Th17) pathway plays an important role in the pathogenesis of atopic dermatitis (AD). Inhibition of the IL-23/Th17 pathway with monoclonal antibodies reduces skin inflammation in animal models. AIM: To investigate the association between IL-17A and IL-23R gene single nucleotide polymorphisms (SNPs) and the development of AD in a Polish population. METHODS: Blood samples were collected from 166 patients with AD and 160 controls. We analyzed two SNPs, -152 G/A IL-17A and 1142 G/A IL-23R, using PCR and restriction fragment length polymorphism (RFLP) analysis. RESULTS: There was no statistically significant difference between the examined IL-17A SNP and the incidence of AD (P > 0.05 for all comparisons). Analysis of the IL-23R gene SNP showed no relationship between AD and the G/A genotype or presence of the A allele. The study did not establish any links between the IL-23R and IL-17A gene SNPs and the likelihood of developing AD resulting from gene-gene interaction. However, there was a significant relationship between the A/A genotype in the -152 G/A IL1-7A SNP and the coexistence of AD and asthma (P < 0.04). Analyzing the association between AD severity and the occurrence of IL-17A SNP, we found that subjects with the A/A genotype were at higher risk of developing moderate or severe AD (P = 0.02). CONCLUSIONS: We found no evidence of any effect of IL-17A or IL-23R SNPs on the occurrence of AD in our Polish population. However, the A/A genotype in IL-17A was found to predispose to increased AD severity and coexistence of AD and asthma.

Use of ionic chromatography in determining the contamination of apple juice by lactic acid.

High-performance anion exchange chromatography (HPAEC) with conductometric detection was used for the determination of the lactic acid content of apple juice subjected to fermentation with the strains of Lactobacillus brevis and Lactobacillus plantarum, obtained from a collection, at 20 and 30 degrees C. At the same time, lactate content was determined by means of enzymatic tests and biosensors. Lactic acid concentrations determined by the chromatographic method are similar to those obtained during analysis by enzymatic tests. However, acid concentrations determined by means of biosensors substantially diverge from these results.

Comparison of three staining techniques for the morphometric study of rainbow trout (Oncorhynchus mykiss) spermatozoa.

This study was designed to compare the performance of the kits Diff-Quick, Hemacolor and Spermac for staining the spermatozoa of rainbow trout. Automated sperm morphology analysis (ASMA) was performed using two image analysis programs to determine the sperm measurements: head size (length, width, area and perimeter), shape (ellipticity, rugosity, elongation and regularity) and tail length. Diff-Quick was found to be the best procedure for staining the trout spermatozoa. The use of this method rendered the highest number of cells correctly analyzed, and provided good colour intensity and contrast of the sperm head. No differences among the methods were detected in terms of tail length measurements. Mean values established using Diff-Quick for the main morphometric variables were: head length 2.93+/-0.13 microm; head width 2.33+/-0.15 microm and tail length 34.16+/-1.66 microm. Based on these findings, we recommend the Diff-Quick staining kit for its accurate and reproducible morphometric results. Notwithstanding, when analyzing the sperm tail of the rainbow trout, the Spermac method offers improved contrast.

Isolation and characterization of alpha1-proteinase inhibitor from common carp (Cyprinus carpio) seminal plasma.

Using a three-step procedure, we purified (79 and 51.6-fold to homogeneity) and characterized the two isoforms (a and b) of alpha1-proteinase inhibitor-like protein from carp seminal plasma. The isoforms have molecular masses of 55.5 and 54.0 kDa, respectively. These inhibitors formed SDS-stable complexes with cod and bovine trypsin, chymotrypsin and elastase. The thirty-three amino acids within the reactive loop SLPDTVILNRPFLVLIVEDTTKSILFMGKITNP were identified for isoform b. The same first ten amino acids were obtained for isoform a, and this sequence revealed 100% homology to carp alpha1-proteinase inhibitor (alpha1-PI) from perimeningeal fluid. Both isoforms of alpha1-PI are glycoproteins and their carbohydrate content was determined to be 12.6 and 12.1% for a and b, respectively. Our results indicated that alpha1-PI is one of the main proteins of carp seminal plasma. Using polyclonal anti-alpha1-PI antibodies, alpha1-PI was for the first time localized to the carp testis. The presence of alpha1-PI in testis lobules and in the area surrounding spermatides suggests that this inhibitor may be involved in the maintenance of testis connective tissue integrity, control of spermatogenesis or protection of tissue and spermatozoa against unwanted proteolysis. Since similar alpha1-PI has been identified in rainbow trout semen it can be suggested that the presence of alpha1-PI in seminal plasma is a common feature of cyprinid and salmonid fish.

Effects of liquid storage on amidase activity, DNA fragmentation and motility of turkey spermatozoa.

Short-term liquid storage of turkey semen is of great interest in the management of turkey reproduction due to the extensive use of artificial insemination. This study examined changes in DNA fragmentation (using a comet assay), sperm motility characteristics (using computer-aided sperm analysis), and amidase activity (using a colorimetric assay) of turkey sperm stored for 24 and 48 h. In addition we found that turkey spermatozoa contain besides acrosin, additional two serine proteinases of molecular weight of 34 and 42 kDa. We found that, after 48 h of liquid storage, decreases in sperm motility characteristics and increases in amidase activity and DNA fragmentation occurred. An increase of amidase activity was found after 24h. Decreases in sperm motility and increase in DNA fragmentation were found after 48 h of storage. These data suggest that a decrease in turkey sperm quality during short-term storage is related to disturbances to the acrosome, presumably related to premature activation of acrosomal serine proteinases, and to a lesser extent a decrease in sperm motility characteristics and damage of sperm DNA.

Effects of UV irradiation and hydrogen peroxide on DNA fragmentation, motility and fertilizing ability of rainbow trout (Oncorhynchus mykiss) spermatozoa.

Preservation of DNA integrity is essential for protection of sperm quality. This study examined, with the use of comet assay, DNA fragmentation of rainbow trout (Oncorhynchus mykiss) spermatozoa subjected to UV irradiation (2,075 microW/cm(2), 0-15 min) or oxidative stress induced by hydrogen peroxide (0-20mM). Sperm motility and fertilizing ability were also measured. A dramatic increase in DNA fragmentation was recorded after 5 min UV irradiation but no significant changes in sperm motility were observed at this time. Longer irradiation resulted in a decrease in motility parameters and further increase of DNA fragmentation. UV irradiation caused a clear decrease in the percentage of eyed embryos and most of the embryos did not hatch. When highly diluted sperm suspensions (50,000-fold) were exposed to 0.1mM H(2)O(2) evident increase in DNA fragmentation was observed. On the other hand, when more concentrated sperm suspensions (diluted only 40-fold) were employed (in order to conduct motility and fertilization measurements at the same time) 1-20mM H(2)O(2) caused only moderate increase in DNA fragmentation and dose-dependent decline in sperm motility and fertilizing ability. This suggests that toxic effects of H(2)O(2) were primarily related to inhibition of sperm motility. Our results demonstrate that comet assay can be used for monitoring the effectiveness of fish sperm DNA inactivation by UV irradiation. Therefore, the comet assay together with sperm motility analysis can be applied in optimization works of gynogenetic procedures in fish. Lack of effectiveness of H(2)O(2) in inducing major DNA fragmentation suggests presence of mechanisms of antioxidative defense in rainbow trout spermatozoa.

Content of iron, copper and zinc in white sugar samples from Polish and other European sugar factories.

White sugar is a very pure food product, even though it contains very small, significant amounts of soluble and insoluble impurities. The content of these impurities has nutritional significance and determines the usefulness of sugar for various industrial applications. The aim was to evaluate the content of iron, copper and zinc in samples of white sugar from Polish factories compared with commercial white sugar samples from other European countries. The investigations were conducted on 72 campaign-averaged white sugar samples from 37 different Polish sugar factories from 1996 to 2000 and on 16 commercial white sugar samples from nine European countries. The content of iron, copper and zinc in those sugar samples was determined by means of FAAS both in the sediment and in the solution after filtration on 0.45- micro m filters of sugar water solution. The content of iron, copper and zinc was low (averages 0.29, 0.06 and 0.07 mg x kg(-1), respectively) in all the white sugar samples from Polish sugar factories and other European countries. Iron and copper found in all white sugar samples were mainly in insoluble form - 77 and 69%, respectively. The contents of water-insoluble iron and water-soluble zinc in white sugar increase with a lowering of the quality of sugar evaluated according to the standards of the EU sugar market regime.

A new procedure for measuring peripheral compression in normal-hearing and hearing-impaired listeners.

Forward-masking growth functions for on-frequency (6-kHz) and off-frequency (3-kHz) sinusoidal maskers were measured in quiet and in a high-pass noise just above the 6-kHz probe frequency. The data show that estimates of response-growth rates obtained from those functions in quiet, which have been used to infer cochlear compression, are strongly dependent on the spread of probe excitation toward higher frequency regions. Therefore, an alternative procedure for measuring response-growth rates was proposed, one that employs a fixed low-level probe and avoids level-dependent spread of probe excitation. Fixed-probe-level temporal masking curves (TMCs) were obtained from normal-hearing listeners at a test frequency of 1 kHz, where the short 1-kHz probe was fixed in level at about 10 dB SL. The level of the preceding forward masker was adjusted to obtain masked threshold as a function of the time delay between masker and probe. The TMCs were obtained for an on-frequency masker (1 kHz) and for other maskers with frequencies both below and above the probe frequency. From these measurements, input/output response-growth curves were derived for individual ears. Response-growth slopes varied from >1.0 at low masker levels to <0.2 at mid masker levels. In three subjects, response growth increased again at high masker levels (>80 dB SPL). For the fixed-level probe, the TMC slopes changed very little in the presence of a high-pass noise masking upward spread of probe excitation. A greater effect on the TMCs was observed when a high-frequency cueing tone was used with the masking tone. In both cases, however, the net effects on the estimated rate of response growth were minimal.

The effect of basilar-membrane nonlinearity on the shapes of masking period patterns in normal and impaired hearing.

Masking period patterns (MPPs) were measured in listeners with normal and impaired hearing using amplitude-modulated tonal maskers and short tonal probes. The frequency of the masker was either the same as the frequency of the probe (on-frequency masking) or was one octave below the frequency of the probe (off-frequency masking). In experiment 1, MPPs were measured for listeners with normal hearing using different masker levels. Carrier frequencies of 3 and 6 kHz were used for the masker. The probe had a frequency of 6 kHz. For all masker levels, the off-frequency MPPs exhibited deeper and longer valleys compared with the on-frequency MPPs. Hearing-impaired listeners were tested in experiment 2. For some hearing-impaired subjects, masker frequencies of 1.5 kHz and 3 kHz were paired with a probe frequency of 3 kHz. MPPs measured for listeners with hearing loss had similar shapes for on- and off-frequency maskers. It was hypothesized that the shapes of MPPs reflect nonlinear processing at the level of the basilar membrane in normal hearing and more linear processing in impaired hearing. A model assuming different cochlear gains for normal versus impaired hearing and similar parameters of the temporal integrator for both groups of listeners successfully predicted the MPPs.

Luteotrophic action of prolactin during the early luteal phase in pigs: the involvement of protein kinases and phosphatases.

Prolactin (PRL) involvement in the regulation of luteal steroidogenesis in pigs during the early luteal phase and pregnancy is well documented. The intracellular mechanism of PRL action in steroidogenic cells, however, is not fully recognized yet. In the current study, we have tested the hypothesis that protein kinase C (PKC) and tyrosine kinases (PTK) as well as serine-threonine (PP) and tyrosine phosphatases (PTP) are involved in PRL signaling in luteal cells originated from the early corpora lutea (CL) of cyclic sows. Luteal cells (50 000 cells/ml M199) were incubated for 8 h (37 degrees C) with PRL (200 ng) and low density lipoproteins (LDL) to stimulate P(4) production. In addition, treatments included: PKC inhibitors--staurosporine and chelerythrine chloride; tyrosine kinase inhibitors--genistein and tyrphostin; serine-threonine phosphatase inhibitors--okadaic acid, cantharidin (inhibitors of PP1/2A) and cypermethrin (inhibitor of PP2B); and tyrosine phosphatase inhibitor--sodium orthovanadate. Moreover, after incubation (37 degrees C) with PRL (200 ng) for 2, 5, 10 or 20 min, luteal cells were homogenized and cytosolic as well as membrane fractions have been obtained. This was followed by partial purification of the subcellular fractions by DEAE-cellulose chromatography and determination of PKC activity by measuring the transfer of (32)P from [gamma-(32)P]ATP to histone III-S. In unstimulated porcine luteal cells the major proportion of PKC activity was present in the cytosol. Incubation of luteal cells with PRL resulted in a rapid, time dependent increase in the amount of PKC activity in the membrane fraction and a decrease in the amount of PKC activity in the cytosol fraction. PKC activity in the membrane fraction was maximal after 5 min of exposure the cells to PRL. Inhibitors of PKC and PTK suppressed PRL and LDL-induced P(4) production by porcine luteal cells. It is of interest that stimulated P(4) production was also reduced by inhibitors of PTP and PP1/2A (okadaic acid, cantharidin). In contrast, cypermethrin did not affect P(4) production stimulated by PRL and LDL. The results of the current study support the hypothesis that PKC and tyrosine kinases are intracellular mediators of PRL action in porcine luteal cells during the first days of the estrous cycle. The involvement of protein phosphatases in transmission of the PRL signal in early luteal cells in pigs is also suggested.

Intensity discrimination and detection of amplitude modulation.

Thresholds for detection of low-rate sinusoidal amplitude modulation and for detection of intensity increments were measured over a wide range of levels in an examination of the relationship between these fundamental aspects of intensity processing. As expected, thresholds measured with a continuous 1-kHz tone decrease with increasing carrier/pedestal level. For levels between 6 and 85 dB SPL the data are well described by 10 log delta I/I = 0.44.(20 log m) + D(fm), where delta I/I is the Weber fraction for increment detection, m is the modulation index at threshold, and D(fm) depends on modulation rate (fm). The relationship between the psychometric functions for modulation and increment detection is also consistent with this equation. The data indicate a clear relationship between modulation and increment detection and thus provide an important additional consideration for models of modulation processing. No existing models provide an adequate account of this relationship.

Effects of fast-acting high-frequency compression on the intelligibility of speech in steady and fluctuating background sounds.

This study examines whether speech intelligibility in background sounds can be improved for persons with loudness recruitment by the use of fast-acting compression applied at high frequencies, when the overall level of the sounds is held constant by means of a slow-acting automatic gain control (AGC) system and when appropriate frequency-response shaping is applied. Two types of fast-acting compression were used in the high-frequency channel of a two-channel system: a compression limiter with a 10:1 compression ratio and with a compression threshold about 9 dB below the peak level of the signal in the high-frequency channel; and a wide dynamic range compressor with a 2:1 compression ratio and with the compression threshold about 24 dB below the peak level of the signal in the high-frequency channel. A condition with linear processing in the high-frequency channel was also used. Speech reception thresholds (SRTs) were measured for two background sounds: a steady speech-shaped noise and a single male talker. All subjects had moderate-to-severe sensorineural hearing loss. Three different types of speech material were used: the adaptive sentence lists (ASL), the Bamford-Kowal-Bench (BKB) sentence lists and the Boothroyd word lists. For the steady background noise, the compression generally led to poorer performance than for the linear condition, although the deleterious effect was only significant for the 10:1 compression ratio. For the background of a single talker, the compression had no significant effect except for the ASL sentences, where the 10:1 compression gave significantly better performance than the linear condition. Overall, the results did not show any clear benefits of the fast-acting compression, possibly because the slow-acting AGC allowed the use of gains in the linear condition that were markedly higher than would normally be used with linear hearing aids.