Application of Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry for Identification of Coagulase-negative Staphylococci Isolated from Milk of Cows with Subclinical Mastitis.

The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81.8%). The most frequent cause of subclinical mastitis was found to be Staphylococcus sciuri (39%), while S. vitulinus was detected in 15% of the milk samples. The results obtained indicate that MALDI-TOF MS can be used for the identification of CNS isolated from bovine mastitis as a method supplementary to biochemical tests.

Applicability of 2D gel electrophoresis and liquid chromatography in proteomic analysis of urine using mass spectrometry MALDI-TOF.

Proteomics including the studies of the structure, function and dependences between proteins is more and more extensively applied in human medicine and veterinary medicine. The analysis of protein profiles of tissues and body fluid from healthy and ill individuals allows to identify diagnostic, prognostic and predictive markers in various pathological states in people and animals. This paper presents preparation of urine samples for analysis in the mass spectrometer MALDI-TOF (Ultraflextreme, Bruker, Bremen, Germany) by means of two methods: liquid chromatography based on the system Nano-LC (PROTEINER FC II, Bruker Daltonics, Bremen Germany). and two-direction electrophoresis 2DE (GE Healthcare, United Kingdom). Both methods enable separation of the mixture under consideration into individual fractions of high purity indispensable for obtaining readable mass spectra. The purpose of this paper is to determine applicability of these methods in analysis of protein composition of urine samples.

The loop-mediated isothermal amplification assay for rapid diagnosis of Babesia canis canis infections in dogs.

The aim of this study was to use a rapid and easy DNA-based test, the loop-mediated isothermal amplification (LAMP), for diagnosis of Babesia canis canis infections in dogs. 10 DNA samples of 18S RNA-A and 10 DNA samples of 18S RNA-B of B. canis canis were used in the study. LAMP method could successfully detect DNA in all examined samples down to 0.1 pg dilution. Obtained results suggest that this method has high specificity and sensitivity and can be applied in analytical laboratories in diagnosis of canine babesiosis.

HPLC Identification of Copper (II)-Trans-Resveratrol Complexes in Ethanolic Aqueous Solution.

Reversed-phase high-performance liquid chromatography assay with the diode array detector was applied for detection of trans-resveratrol complexes with copper (II). Two complexes with copper to resveratrol ratio 3:2 and 1:1 were identified in ethanolic-aqueous solutions in neutral and acidic conditions. The matrix-assisted laser desorption ionization-time of flight mass spectrometry was used for evaluation of complexes in eluate liquid fraction. The structures of complexes were modeled by Titan, Spartan and HyperChem software. The findings obtained satisfactorily explain the chromatographic data and could provide useful an additional data about these forms in which is present trans-resveratrol in wine (free and/or forming complex with copper).

Peripheral mechanisms of intestinal dysmotility in rats with salsolinol induced experimental Parkinson's disease.

Gastrointestinal dysmotility in Parkinson's disease (PD) has been attributed in part to peripheral neurotoxine action. Our purpose was the evaluation of the salsolinol effect on intramuscular interstitial cells of Cajal (ICC), duodenal myoelectrical activity (DMA) and vagal afferent activity (VAA) in rats with experimental PD. Twenty rats were divided into 2 equal groups. Experimental PD was produced in one group by 3 weeks of the intraperitoneal salsolinol injections (50 mg/kg/day), whereas the 2-nd group served as control. DMA and VAA were recorded in both groups during fasting and stepwise--gastric distension (GD) of 10 ml. Subsequently fragments of duodenum were removed and intramuscular ICC were assessed as c-Kit antigen percentage in the duodenal muscular zone. Analyses of the fasting DMA and VAA recordings didn't reveal differences between the compared groups. During GD increase of DMA dominant frequency (p=0.04) and VAA frequency (p<0.01) was observed in the controls whereas in the salsolinol group both parameters remained unchanged. Image analysis of duodenum revealed decreased c-Kit expression in the salsolinol-injected animals (p=0.05). The results of our study may suggest the direct effect of salsolinol on both ICC and neuronal pathways of gastro-duodenal reflexes.

Peripheral mechanisms of intestinal dysmotility in the morphine tolerant and dependent rats.

Changes of intestinal motility and transit produced by tolerance to and dependence upon morphine have been partly attributed to peripheral mechanisms. We evaluated the effect of chronic peripheral morphine administration and peripheral mu-receptor blockade on vagal afferent activity (VAA) and c-Kit positive intramuscular cells of Cajal (ICCs). Ten rats were subjected to chronic subcutaneous morphine infusion for 72 h with subsequent VAA recording. Potential frequency was evaluated within recordings before and after mu receptor blockade by (D)-Phe -Cys -Tyr -(D)-Trp -Orn -Thr -Phe -Thr (CTOP) i.p. injections. Afterwards the rats were sacrificed and intramuscular c-Kit antigen expression was assessed by image analysis within removed fragments of duodenum and ascending colon. An equal group of rats served as a control for VAA and c-Kit expression. Analysis of VAA revealed similar frequencies of potentials in morphine tolerant / dependent rats before CTOP and in the controls. CTOP increased potential frequency in the morphine group which effect was visible mostly within the first 20 minutes (p=0.01). The morphine infused animals presented also higher c-Kit expression in both the duodenum (p<0.001) and the ascending colon (p<0.001) in comparison to the control group. Results of our study may indicate the involvement of both the intestinal wall and the long vago-vagal reflexes in tolerance to and dependence upon opioids.

[Effects of antibiotic therapy on endoscopic examination of the upper gastrointestinal tract in children with Helicobacter pylori infections]. / Wplyw antybiotykoterapii na obraz zmian endoskopowych górnego odcinka przewodu pokarmowego u dzieci zakazonych Helicobacter pylori.

The aim of the study was defining the antibiotic influence for endoscopic picture of the upper segment of gastrointestinal tract in Helicobacter pylori infection. We remained retrospectively analysis of 790 endoscopies. In 62 of the analysed patients we found Helicobacter pylori infection. We divided these children into two groups. The first group, contained 15 children, who received antibiotics for the previous year, for any reasons. The second, comparative group was made up of 47 children, who didn't receive antibiotics. We compared 18 characteristics of endoscopic picture in the oesophagus, stomach and duodenum mucosa. As a result of this study we conclude, that: 1) the most frequently the endoscopic changes were found in the stomach, as an erythema, an oedema and a nodularity of mucosa; 2) the inflammatory changes were in oesophagus and duodenum, as well; 3) we didn't observe the clear influence of antibiotics on the development of endoscopic changes in Helicobacter pylori infection; 4) in children, who didn't receive antibiotics, we observed polypus in the oesophagus, erosions in the stomach and ulcer in the duodenum. This may suggest the preventive role of antibiotics on intensity of gastrointestinal changes in Helicobacter pylori infected children.

The effect of the year angiotensin-converting enzyme inhibitors (ACE I) intake on circadian heart rate variability in patients with primary hypertension.

INTRODUCTION: The autonomic nervous system (ANS) disturbances have been considered as one of the important factors in development of essential hypertension. However information about the effect of antyhypertensive treatment (angiotensin-converting enzyme inhibitors--ACEI) on cardiac ANS activity is ccscarce. AIM: The purpose of the study was to evaluate the HRV circadian changes in patients with essential hypertension, treated with ACEI throughout the period of one year. METHODS: Ten patients with essential hypertension, aged 26 to 64 years (mean 54.3 +/- 12.3) and 10 healthy volunteers, matched for age and gender were qualified for the investigation. Twenty four-hour arterial blood pressure measurements with simultaneous ECG monitoring were conducted in both mentioned groups. The hypertensive patients were examined before and after 1 year of ACE inhibitor (enalapril) intake. HRV analysis was carried out by the fast Fourier transformation (FFT). The time and spectral parameters were compared in 8 examined subjects characteristic for dippers, in time periods of day (6 a.m.-10 p.m.) and night (10 p.m.-6 a.m.). RESULTS: The results obtained before the treatment, likewise in control group, presented night's increases of RMSSD (p < 0.05), pNN50 (NS, p > 0.05), VLF, LF (p < 0.05) and HF (NS) mean values with a decrease of LF/HF ratio (p < 0.05). Similar tendencies in the cardiac ANS activity were observed after 1 year of the enalapril intake. A comparison of day's HRV before and after one year of the ACE inhibition presented the increase in values of the parasympathetic system reflecting parameters: RMSSD, pNN50 (p = 0.01) and HF (NS) with a decrease of LF/HF ratio (p < 0.01). The evaluation of the night's HRV parameters presented the increase of HF mean value (p < 0.05), without any other significant changes between the trials. CONCLUSIONS: After 1-year of ACE inhibitor administration the only improvement of the day's parasympathetic activity was observed. The night's vagal activity suggested as one of the essential factor in development of hypertension remained unchanged in patients qualified into "dippers" hypertension group.

[Autonomic nervous system and gastric myoelectric activities in type 1 and type 2 diabetes]. / Aktywnosc autonomicznego ukladu nerwowego a czynnosc mioelektryczna zoladka w cukrzycy typu 1 i 2.

The aim of the study was to compare autonomic function and gastric myoelectric activity in patients with type 1 and 2 of diabetes. We used the skin electrodes for recording gastric myoelectric activity (EGG) and the heart rate variability (HRV). Patients were divided into two groups. The first group: 19 patients with IDDM. The second-16 patients with NIDDM. We made two comparative control groups, matched with age and gender. We conclude, that decrease in gastric myoelectric activity was observed in both groups of patients, mostly in 1 group (IDDM) of patients. There was increase of heart rate and reduction of spectral analysis parameters in both types of diabetic. In patients with type 1 we noticed the increase of sympathetic activity (LF/HF ratio increased). In patients with type 2 there was the non significant decrease in sympathetic activity (LF/HF ratio decreased). On the basis of our observations we confirmed the correlation among autonomic function tested by HRV and changes of stomach electric activity. Decrease in HRV and gastric myoelectric activity could be indicating damage of parasympathetic system in IDDM.

[The effect of aging on the activity of the autonomic nervous system in long distance runners]. / Wplyw starzenia sie na aktywnosc autonomicznego ukladu nerwowego u biegaczy dlugodystansowych.

The aim of the study was to evaluate the effect of endurance physical training on the autonomic function in runners. The study was performed on 20 long distance runners, divided into two groups. The first group of runners contained 12 young men of age of 20-24 years (age 21.75 +/- 1.4 years: mean S.D.--standard deviation). Into the second group 9 experienced runners, who practice long distance running for over 40 years, were qualified. The mean age of those runners was 52.9 +/- 7.2 years. Moreover two control groups, matched for age and gender were recruited. One of them was made up of 12 healthy volunteers (age 21.6 +/- 1.23 years), the second contained 9 men (age 52.9 +/- 7.2 years). In this study, the measurement of the cardiac autonomic nervous system, using the HRV computer assisted programme (Proster, Poland), was performed. The characteristics of the HRV (heart rate variability) were estimated. We have observed, that the RR intervals in runners were longer than in healthy volunteers (1033.5 +/- 185.4 ms vs. 888.1 +/- 115.3 ms. p < 0.01). We have observed a tendency towards increasing of the RR intervals with the increase of age, of the tested person, as well. A reverse tendency, a shortening of the RR intervals with ageing, was observed in the control groups. The standard deviation of the RR intervals in runners was higher than in the control group (p < 0.05). In the young runners we noticed the prevalence of parasympathetic activity, expressed by the HF (high frequency) spectra (875.7 ms2 vs. 816.7 ms2) in comparison with the healthy individuals. In the old runners the prevalence of sympathetic activity, expressed by LF (low frequency) spectra (1088.75 ms2 vs. 220.1 ms2, p < 0.05) and parasympathetic activity (920.38 ms2 vs. 294.25 ms2) in comparison with the control group, was observed. It was expressed by the increase of the LF/HF ratio (0.75 vs. 2.1). The results of our study indicate, that the autonomic, particularly sympathetic, activity in sportsmen is not affected by ageing up to the sixth decade of life.

[Myoelectric activity of the stomach and esophageal pH changes in reflux disease]. / Aktywnosc mioelektryczna zoladka a zmiany pH w przelyku w chorobie refluksowej.

UNLABELLED: The oesophageal inflammation during gastrointestinal reflux disease (GERD) has been considered to be an acid and pepsine related disorder. The oesophagus remains under the prolonged exposure of gastric acid, which damages the mucosa and causes an esophagitis. Amongst the most important pathogenetic factors of GERD, the delayed gastric emptying and transient episodes of low esophageal sphincter (LES) relaxation have been described. The examinations like electrogastrography (EGG) and 24-hours pH-metry are useful for the evaluation of these disturbances. AIM: The aim of the study was to confirm the co-existence among the abnormal gastric myoelectric activity and the esophageal pH changes in patients with GERD. METHODS: Ten patients (8 m., 2 f.) 20-61 years of age (mean 46 +/- 15.3 years) with GERD were examined. The EGG recording lasted one hour: 30 min. in fasting state and 30 min. after a standard meal. The 24-hours pH-metry measurement, five cm above the upper edge of LES, was done on separate days. A control group, matched for number and age, for the comparison of the EGG recording, was recruited. RESULTS: Amongst the patients suffering from GERD the results of fasting EGG showed the lower normogastria (mean 45.3 +/- 21.9% vs. 86.5 +/- 8.8%: p < 0.01) and the high dysrrhytmia time period (mean 54.7 +/- 21.9% vs. 13.5 +/- 9.6%: p < 0.01), than in the control group. After the meal, the dysrrhytmia persisted (mean 43.5 +/- 24.3% vs. 9.6 +/- 11.5%: p < 0.01). The mean increase of the slow waves amplitude after the meal was 0 +/- 55% vs. 80 +/- 146%; p < 0.05. The recording of the 24-hours pH-metry showed an increase of the pH < 4 episodes number (mean 329 +/- 246) and time percentage (mean 12.2 +/- 6.6%). The patients obtained mean 65.5 +/- 61 points in the De Meester and Johnson scoring. A significant correlation among the postprandial dysrrhytmia and reflux episodes time percentages was observed (p < 0.05). CONCLUSIONS: The results of our studies suggest the importance of the gastric myoelectric activity disturbances in pathogenesis of GERD.

[The effect of chronic alcohol use on heart rate variability]. / Wplyw przewleklego spozycia alkoholu na zmiennosc rytmu zatokowego serca.

A long lasting alcohol intake causes, amongst numerous systemic damages, also the autonomic nervous system (ANS) dysfunction, which causes the autonomic heart rate regulation disorders. The aim of the study was to evaluate the autonomic regulation of the circulation in chronic alcoholism. Seventeen alcoholics, 24-55 years of age (mean 43 +/- 5.2 years) were examined. They have been abstainers for 2-6 years. The cardiac ANS function was evaluated using the HRV measurement. The HRV was registered using V6 EKG lead. The recording was performed through the 15 min of resting conditions and 5 min of the deep breathing test. A group containing healthy volunteers, matched for age and gender, for the comparison of the HRV results was recruited. In the examined group, during the resting conditions, the significant RR period changes weren't observed (999.7 +/- 139.2 vs. 967 +/- 144.9; p > 0.05). The nonsignificant lower values of the spectral analysis parameters of HRV: LF (954.1 +/- 1162.6 vs. 1456.4 +/- 1327.1; p > 0.05) and HF (676.4 +/- 414.2 vs. 1557 +/- 1854.4; p > 0.05) and LF/HF ratio (1.5 +/- 1.14 vs. 1.38 +/- 1.28; p > 0.05) were also noticed. In response to the DB test, the mean value of the RR period wasn't significantly changed (921.4 +/- 152.3 vs. 930.6 +/- 137.8; p > 0.05). In DB test the significant decrease of LF (3465.8 +/- 2750.1 vs. 11558.6 +/- 7902.5; p < 0.001) and HF (406.1 +/- 366.8 vs. 1665 +/- 1757.1; p < 0.01) was observed. No significant change of LF/HF mean ratio (11.6 +/- 6.97 vs. 14.7 +/- 11.6; p > 0.05) was noticed. The results of our study indicate on the maintenance of the HRV disorders in chronic alcoholics, during the abstinence.

Isolation and characterization of two antigens of Corynebacterium hofmannii.

Banach, T. M. (University of Manitoba, Winnipeg, Canada), and R. Z. Hawirko. Isolation and characterization of two antigens of Corynebacterium hofmannii. J. Bacteriol. 92:1304-1310. 1966.-A serologically active substance, extracted from sonically treated cells of Corynebacterium hofmannii with hot HCl, produced two precipitin lines by immunodiffusion tests with a hyperimmune homologous serum. Extracts of other species failed to precipitate with the hofmannii antiserum. The active fraction was eluted from a diethylaminoethyl cellulose column in the third adsorption peak at a linear concentration of 0.5 m KCl, and produced two precipitin lines which corresponded in identity to those formed by the acid extract. Separation of the antigens was achieved by rechromatography on a Sephadex G-200 column; the major antigen was designated A; the minor, B. The homogeneity and purity of each antigen was established by immunoelectrophoresis and, in addition, that of antigen A by disc electrophoresis. Biochemical analyses showed that both antigens were composed of a major protein component with polysaccharide and nucleic acid present in an approximate ratio of 17:3:1, respectively. Glutamic acid, aspartic acid, alanine, glycine, valine, and leucine were the main amino acids present. Antigen A contained 17% less protein and 3.5% less carbohydrate than antigen B. The principal sugars of antigen A were identified as arabinose and glucose. The molecular weight, estimated by gradient centrifugation, was 16,500 for antigen A and 21,000 for antigen B.