The occurrence of endotoxin in sows with coliform mastitis

The purpose of this study was to determine the role of endotoxins in the etiology of coliform mastitis /CM/ in sows under field conditions, by using the Limulus amebocyte lysate /LAL/ test for the detection of endotoxins in the blood of pigs. For this purpose, blood samples from 40 healthy sows and 46 sows with clinical signs of CM were drawn once between 24 and 72 h post partum and tested accordingly. Only one clinically healthy sow 2.5% showed the presence of endotoxins in the serum. In sows clinically affected with one or more symptoms of the CM, the bacterial toxins were detected in 15 cases /32.5%/. The results of these studies support the observation that in only a certain percentage of all CM cases the occurrence of endotoxin in blood can be shown. Sampling time in relation to the occurrence of clinical symptoms and sampling frequency may have had an influence on the detectability of circulating endotoxins. Consequently, not all cases with endotoxemia may have been identified. The usefulness of the LAL test for endotoxin detection in the blood of pigs was confirmed.     

Automatic Detection of Whole Night Snoring Events Using Non-Contact Microphone

Objective

Although awareness of sleep disorders is increasing, limited information is available on whole night detection of snoring. Our study aimed to develop and validate a robust, high performance, and sensitive whole-night snore detector based on non-contact technology.

Design

Sounds during polysomnography (PSG) were recorded using a directional condenser microphone placed 1 m above the bed. An AdaBoost classifier was trained and validated on manually labeled snoring and non-snoring acoustic events.

Patients

Sixty-seven subjects (age 52.5±13.5 years, BMI 30.8±4.7 kg/m², m/f 40/27) referred for PSG for obstructive sleep apnea diagnoses were prospectively and consecutively recruited. Twenty-five subjects were used for the design study; the validation study was blindly performed on the remaining forty-two subjects.

Measurements and Results

To train the proposed sound detector, >76,600 acoustic episodes collected in the design study were manually classified by three scorers into snore and non-snore episodes (e.g., bedding noise, coughing, environmental). A feature selection process was applied to select the most discriminative features extracted from time and spectral domains. The average snore/non-snore detection rate (accuracy) for the design group was 98.4% based on a ten-fold cross-validation technique. When tested on the validation group, the average detection rate was 98.2% with sensitivity of 98.0% (snore as a snore) and specificity of 98.3% (noise as noise).

Conclusions

Audio-based features extracted from time and spectral domains can accurately discriminate between snore and non-snore acoustic events. This audio analysis approach enables detection and analysis of snoring sounds from a full night in order to produce quantified measures for objective follow-up of patients.     

Genetic structure of local Ukrainian cattle breeds

Analysis of 6 cattle breeds (5 local Ukrainian breeds and the Holstain breed) on the 9 polymorphous molecular-genetic markers (transferrin, ceruloplasmin, amylase-1, posttransferrin, receptor to vitamin D, haemoglobin, leptin, kapa-casein) was carried out. The rare allele of transferrin was revealed in two local breeds and the rare allele of leptin--in two another local breeds. Associations between syntenic loci (transferrin, ceruloplasmin, kappa-casein, receptor to vitamin D) were observed only in the local breeds but not in the Holstain one. Locus-specific conservation of the ancestor allelic variants in the local breeds and effect of selection on interloci associations are discussed.     

Participation of structural gene markers and anonymous DNA sequences in genetic differentiation in the species Ovis aries L. and Ovis nivicola borealis

Genetic variability and genetic differentiation in two Ovis species--domesticated (Askanian sheep, Sokilska and Kulunda sheep) and wild (bighorn sheep) were analyzed using different types of molecular-genetic markers--genetic-biochemical (30 loci) and DNA (ISSR-PCR) ones. High level of genetic variability was revealed. The average heterozygosity of biochemical markers loci was in range of 0.073-0.188. Markers involved into process of genofond divergence of two closely related species were revealed.     

Sleep/Wake Dynamics Changes during Maturation in Rats

Objectives

Conventional scoring of sleep provides little information about the process of transitioning between vigilance states. We applied the state space technique (SST) using frequency band ratios to follow normal maturation of different sleep/wake states, velocities of movements, and transitions between states of juvenile (postnatal day 34, P34) and young adult rats (P71).

Design

24-h sleep recordings of eight P34 and nine P71 were analyzed using conventional scoring criteria and SST one week following implantation of telemetric transmitter. SST is a non-categorical approach that allows novel quantitative and unbiased examination of vigilance-states dynamics and state transitions. In this approach, behavioral changes are described in a 2-dimensional state space that is derived from spectral characteristics of the electroencephalography.

Measurements and Results

With maturation sleep intensity declines, the duration of deep slow wave sleep (DSWS) and light slow wave sleep (LSWS) decreases and increases, respectively. Vigilance state determination, as a function of frequency, is not constant; there is a substantial shift to higher ratio 1 in all vigilance states except DSWS. Deep slow wave sleep decreases in adult relative to juvenile animals at all frequencies. P71 animals have 400% more trajectories from Wake to LSWS (p = 0.005) and vice versa (p = 0.005), and 100% more micro-arousals (p = 0.021), while trajectories from LSWS to DSWS (p = 0.047) and vice versa (p = 0.033) were reduced by 60%. In both juvenile and adult animals, no significant changes were found in sleep velocity at all regions of the 2-dimensional state space plot; suggesting that maturation has a partial effect on sleep stability.

Conclusions

Here, we present novel and original evidence that SST enables visualization of vigilance-state intensity, transitions, and velocities that were not evident by traditional scoring methods. These observations provide new perspectives in sleep state dynamics and highlight the usefulness of this technique in exploring the development of sleep-wake activity.     

Multiplexing analysis of the polyspecific intrathecal immune response in multiple sclerosis

Intrathecal synthesis of the antibodies specific to neurotrofic viruses: measles (M), rubella (R), Varicella-Zoster (Z), and/or H. simplex (H), known as "MRZH-reaction" plays important diagnostic role in multiple sclerosis (MS). Whereas the analysis of the oligoclonal IgG bands provides high sensitivity, the MRZH-reaction shows high specificity, and hence these methods complement each other. For the first time we applied multiplexing bead-based technology to simultaneously analyze cerebrospinal fluid (CSF) and serum concentrations of antibodies against these viruses, and to calculate the antibody specific indices (ASI's). The method shows reasonable precision: intra-assay, 2.9-6.7%, and inter-assay, 2.0-3.2%. The results are comparable with these obtained with other methods (ELISAs), including two runs of the certified external quality control schemes. Eighty-one percent of the MS cases (n=27) and none of the sex- and age-matched controls (n=14), except one subject with "borderline" anti-measles ASI of 1.5, showed intrathecal synthesis of IgG against at least one of the viruses discussed. The ratios of the MRZH-positive cases in the MS group were: 12/22 for M, 12/19 for R, 13/26 for Z, and 7/26 for H. We conclude that the multiplexing technology can be applied as a tool to study the intrathecal immune response in the diagnosis of MS.     

A novel dimeric dipeptide mimetic of the BDNF selectively activates the MAPK-Erk signaling pathway

On the basis of the structure of beta-turn of loop 2 of brain-derived neurotrophic factor (BDNF), its new dimeric dipeptide mimetic bis-(N-hexanoyl-L-seryl-L-lysine) hexamethylenediamide (GTS-201) was created. It activated TrkB and Erk, did not activate Akt, and exhibited neuroprotective activity in vitro at concentrations of 10^–5–10^–8 M. Unlike the mimetics that activate Erk and Akt, GTS-201 did not exhibit antidepressant properties. For the manifestation of the antidepressant activity of BDNF mimetics, the activation of its both major signaling pathways is required.     

The effect of new non-cross resistant antitumour agents on the energy state of human erythrocytes

Multidrug resistance (MDR) of tumour cells is related to the overexpression of ATP-dependent pumps responsible for the active efflux of antitumour agents out of resistant cells. Benzoperimidine and anthrapyridone compounds exhibit comparable cytotoxic activity against sensitive and MDR tumour cells. They diffuse extremely rapidly across the plasma membrane and render the ATP-dependent efflux inefficient. Such uptake could disturb an energy metabolism of normal cells possessing an elevated level of ATP-dependent proteins, especially erythrocytes having a high level of the MRP1, MRP4 and MRP5 proteins. In this study the effect of five antitumour agents: benzoperimidine (BP1), anthrapyridones (CO1, CO7) and reference drugs used in the clinic: doxorubicin (DOX) and pirarubicin (PIRA), on the energetic state in human erythrocytes has been examined. These compounds have various types of structure and kinetics of cellular uptake (slow--DOX, CO7, moderate--PIRA, fast--BP1, CO1) resulting in their different ability to saturate ATP-dependent transporters. The energetic state of erythrocytes was examined by determination of purine nucleotide contents (ATP, ADP, AMP), NAD(+) and values of adenylate energy charge (AEC) using an HPLC method. It was found that the level of nucleotides as well as the AEC value of erythrocytes were not changed during 24 h of incubation with these agents independently of their structure and ability to saturate ATP-dependent pumps. This is a very promising result in view of their potential use in the clinic as antitumour drugs against multidrug resistant cancers.     

Extracellular ATP induces hyperpolarization and motility stimulation of ciliary cells.

Cellular membrane potential and ciliary motility were examined in tissues cultures prepared from frog palate and esophagus epithelia. Addition of micromolar concentrations of extracellular ATP caused membrane hyperpolarization and enhanced the beat frequency. These two effects of ATP were 1) dose dependent, reaching a maximum at 10 microM ATP; 2) dependent on the presence of extracellular Ca2+ or Mg2+; 3) insensitive to inhibitors of voltage-gated calcium channels; 4) abolished after depleting the intracellular Ca2+ stores with thapsigargin; 5) attenuated by quinidine (1 mM), Cs+ (5-20 mM), and replacement of extracellular Na+ by K+; 6) insensitive to charybdotoxin (5-20 nM), TEA (1-20 microM), and apamin (0.1-1 microM); 7) independent of initial membrane potential; and 8) unaffected by amiloride. In addition, extracellular ATP induced an appreciable rise in intracellular Ca2+. Addition of thapsigargin caused an initial enhancement of the ciliary beat frequency and membrane hyperpolarization. These results strongly suggest the involvement of calcium-dependent potassium channels in the response to ATP. The results show that moderate hyperpolarization is closely associated with a sustained enhancement of ciliary beating by extracellular ATP.     

Low stimulation of NADH oxidation and oxygen consumption by 5-iminodaunorubicin and its derivatives

NADH oxidation and oxygen consumption mediated by 5-iminodaunorubicin and its derivatives have been studied. Experiments were carried out using two enzyme system: microsomes isolated from rat liver and cytochrome c-reductase. All 5-iminoanthracyclines examined were relatively poor electron transfer mediators. In addition, 5-imino-derivatives of daunorubicin modified at sugar moiety were less effective in stimulating NADH oxidation and oxygen radical production than 5-iminodaunorubicin itself. This may be due to the additive effect of 5-imino structure and the presence of bulky substituents at sugar moiety, which might make the compounds poor enzyme substrates.