Maria Szpetnar

Activity of MMP1 and MMP13 and Amino Acid Metabolism in Patients with Alcoholic Liver Cirrhosis

Background Alcoholic liver disease remains one of the most common causes of chronic liver disease worldwide. The aim of this study was to assess the usefulness of metalloproteinases (MMP1 and MMP13) as diagnostic markers of alcoholic liver disease and to determine the changes in free amino acid profile in the patients with alcoholic liver cirrhosis. Material/Methods Sixty patients with alcoholic liver cirrhosis treated in various hospitals of the Lublin region were randomly enrolled. The control group consisted of 10 healthy individuals without liver disease, who did not drink alcohol. Additionally, a group of alcoholics (22 persons) without liver cirrhosis was included in the study. The activity of MMP-1 and MMP-13 in blood plasma of patients and controls was measured using the sandwich enzyme immunoassay technique with commercially available quantitative ELISA test kits. Amino acids were determined by automated ion-exchange chromatography. Results No significant differences were observed in the activity of MMP-1 in alcoholics with or without liver cirrhosis or in controls. Increased serum MMP-13 was found in patients with liver cirrhosis (stage A, B, C) compared to the control group. Patients with alcoholic liver cirrhosis (stage A, B, C) demonstrated reduced concentrations of glutamic acid and glutamine compared to the control group. Plasma levels of valine, isoleucine, leucine, and tryptophan were significantly lower in patients with alcoholic liver cirrhosis (stage C) than in controls. Conclusions MMP-13 can be useful to confirm the diagnosis of alcoholic liver cirrhosis, but levels of MMP-1 are not significantly increased in patients with liver cirrhosis compared to controls. The serum branched-chain amino acid (BCAA) is markedly reduced in patients with stage C alcoholic liver cirrhosis.

Antioxidants in patients receiving total parenteral nutrition after gastrointestinal cancer surgery.

Total parenteral nutrition (TPN) is essential for patients with postoperative impairing gastrointestinal function who are unable to receive and absorb oral/enteral feeding for at least 7 days. Oxidative stress plays a major role in the ethiopathogenesis of cancers. In this study, total antioxidant status (TAS), glutathione peroxidase (GPx), superoxide dismutase, malondialdehyde and ascorbic acid were studied in patients operated because of small intestine, colorectal or pancreatic cancer and subsequently receiving TPN in comparison with patients receiving standard nutrition after the operation. TAS level and GPx activity were decreased in patients with small intestine cancer but did not differ in patients with colorectal and pancreatic cancer before and after surgery. In all patient groups receiving TPN, superoxide dismutase activity after the surgery was kept at the same level as before. On the fifth day after the surgery, malondialdehyde concentration in each group was restored to the value observed before surgery. On the fifth day of TPN treatment, ascorbic acid concentration was increased in every group of patients.

Interrelationships between Morphological, Densitometric and Mechanical Properties of Eggs in Japanese Quails (Coturnix Japonica)

Eggshell quality in birds results from mineral density and composition determining its mechanical endurance. The aim of the study was to determine interrelationships between morphological, densitometric and mechanical properties of eggs in Japanese quails. Twenty four eggs randomly collected from 17-week-old quails were subjected to morphological, denstiometric and mechanical evaluation using dual-energy X-ray absorptiometry (DEXA), quantitative computed tomography (QCT) and three-point bending test. Weight, height and width of eggs were positively correlated with the densitometric parameters obtained using DEXA (egg mineral density (EMD) and egg mineral content (EMC)) and QCT (total egg volume (TEvol) and total eggshell volume (TESvol)). Positive correlations were stated between TEvol and TESvol (r=0.52; P<0.05) and EMD and EMC r=0.83; P<0.05). Egg mineral density revealed positive correlations with TEvol and mean volumetric eggshell mineral density (MvESMD), while EMC was positively correlated with TEvol, TESvol and MvESMD (all P<0.05). Eggshell breaking strength was positively correlated with MvESMD (r=0.53; P<0.05) and negatively correlated with eggshell thickness (r=−0.50; P<0.05). In conclusion, the results obtained in this study showed numerous interrelationships between morphological, densitometric and mechanical properties of eggs in Japanese quails. Both DEXA and QCT were shown to be valuable tools for evaluation of whole egg and eggshell quality with superior prognostic value of QCT for eggshell mechanical endurance prediction. The elaborated experimental model may serve for further investigations on physiological, pharmacological, environmental, nutritional and toxicological factors influencing egg quality.

The fluctuation of free amino acids in serum during acute ischemic stroke

Abstract Currently, little data exists regarding the involvement of free amino acids (AA) in the pathogenesis of ischemic stroke (IS). Thus, our objective was to study the degree of the degree of fluctuation of free amino acids level in serum during the acute phase of IS. The study consisted of eighteen patients (female/male: 10/8; age: 73.1 ± 4.1) with acute IS that was confirmed by way of computed tomography, while twelve sex and age matched individuals were assigned as control group. During the study period, the patients did not receive any supplemental amino acids therapy that could affect the obtained results. The venous blood was obtained after >3 hours fasting at two time-points; time-point 1 – at admission to the hospital; time-point 2 – on day 5 from stroke onset. The blood for control purposes was collected only once, and the blood collection at time-point 1 was done before thrombolytic treatment (nine patients). The amino acids were identified using the Amino Acids Analyser (AAA 400) by INGOS Corp., Praha, Czech Republic. Our results revealed a statistically significant increase of glutamate, cystine and methionine on day 1 of stroke, in comparison to control, whereas, proline level was decreased on day 1 of stroke – in comparison to control serum. On comparing day 5 to the initial day of IS, elevation was observed of levels of asparagine, glycine, tyrosine, arginine, threonine, valine, leucine and phenylalanine. It can be said, then, that ischemic stroke induces both essential and nonessential amino acid fluctuations. Moreover, the decrease in proline and glutamine serum level with the simultaneous increase in the concentration of branch chain amino acids, Glu and Thr suggests a violent mobilization of the body’s proteins. Thus, a decrease of Pro and a simultaneous increase of Glu serum level could be considered as a marker of acute IS.

Thrombolytic treatment decreases glutamate/GABA ratio in serum during acute ischaemic stroke: a pilot study

Abstract There is no information about possible effect of recombinant tissue plasminogen activator (rtPA) therapy on excitotoxic/neuroprotective amino acids during acute phase of ischaemic stroke (IS). Our purpose was to evaluate iv thrombolytic treatment on glutamate (Glu) and gamma-aminobutyric acid (GABA) serum levels during acute IS. Eleven thrombolytic (rtPA group) and 12 non-thrombolytic (non-rtPA group) patients with acute IS were enrolled. The serum samples were obtained at three time points for rtPA group (time point 0: first to fourth hour of stroke; time point 1: immediately after rtPA administration; time point 2: on days 5–7 from stroke onset). The remaining patients had blood collection at two time points: time point 1: 5th–10th hour of stroke and time point 2: on days 5–7 of stroke. Glutamate and GABA were determined by the automated ion-exchange chromatography using Amino Acids Analyser (AAA 400) by INGOS Corp., Praha, Czech Republic. The statistically significant elevation of GABA serum level was noticed directly after thrombolysis (time point 1) in comparison to the corresponding time point in non-rtPA group [0.016 (0.002–0.032) μM/ml vs 0.001 (0.001–0.004) μM/ml for rtPA vs non-rtPA groups, respectively, median (first to third quartile), P < 0.05]. At the same time point, the Glu/GABA ratio was significantly decreased in rtPA group (P < 0.05) suggesting the decrease of excitotoxicity biomarkers in the blood after thrombolysis. Considering the beneficial effect of GABA receptor agonists, the elevation of GABA by rtPA should bring an additional positive features of thrombolytic treatment.

Content of selected amino acids in the gastrocnemius muscle during experimental hypothyroidism in rats

Abstract Introduction: Thyroid hormones affect protein turnover, and in the case of hypothyroidism a decrease in protein synthesis and reduced release of certain amino acids from skeletal muscles are observed. Changes in the amino acid system of skeletal muscles may be responsible for the occurrence of muscle disorders. Material and Methods: The study measured the content of selected amino acids in the gastrocnemius muscle of Wistar rats during experimental hypothyroidism induced by oral administration of methimazole at a concentration of 0.05% in drinking water for 90 d. The rats were divided into four groups: E1 (n = 6) - experimental males, E2 (n = 6) - experimental females, C1 (n = 6) - control males, and C2 (n = 6) control females. Results: A statistically significant reduction occurred in leucine, isoleucine, and 1-methylhistidine levels in males, and 1-methylhistidine in females, in comparison to the control groups. Conclusion: The hypothyroidism-induced changes in amino acid content may be responsible for the occurrence of skeletal muscle function disorders.

Effect of Uterine Inflammatory Status as well as Calcium and Magnesium Concentrations on the Uterine Involution Process in Dairy Cows

Abstract The aim of the study was a comparative analysis of uterine horn diameter, calcium and magnesium concentrations in blood serum and uterine washings in cows both with and without endometritis. The study was performed on 28 cows at 5, 22 and 42 DP (day post partum), of which 14 cows had been diagnosed with endometritis and the other 14 had no endometritis. The animals were selected based on clinical symptoms and cytological examination. Uterine horn diameter was determined in both examined groups of cows using an ultrasonographic device. The level of calcium and magnesium in the blood serum and uterine washings, and protein in uterine washings was determined using a BS-160 Mindray analyzer. Uterus diameter was similar in both examined groups of cows only at 5 DP, and on subsequent days of examination this parameter was higher in the endometritis group compared to the control (P<0.001). Calcium levels in the blood serum of cows with endometritis were lower than in cows without endometritis, both at 22 DP (P<0.001) and at 42 DP (P=0.004). The levels of calcium, magnesium and total proteins in uterine washings were higher in cows with endometritis compared to the group without endometritis, both at 22 and at 42 DPP (P<0.001). The results of this study confirm that the process of uterine involution is slower in cows with endometritis compared to cows without endometritis, and the myometrium of inflamed uterus does not utilize sufficient amounts of calcium and magnesium, which means that the concentrations of these elements in uterine fluid are higher than in cows without endometritis.

Serum Concentration of Free Amino Acids in Dogs Suffering From Perianal Tumours

Abstract The aim of the study was to determine serum free amino acid concentration in male dogs suffering from benign and malignant perianal tumours. Serum concentrations of cysteic acid, glutamic acid, glutamine, aspartic acid, glycine, γ-aminobutyric acid, tryptophan, methionine, arginine, taurine, threonine, tyrosine, alanine, valine, isoleucine, and leucine were significantly changed in dogs suffering from benign and/or malignant tumours when compared to the control group (P<0.05). Serum concentration of serine, phenylalanine, lysine and histidine was not influenced in dogs with neoplastic disease (P>0.05). The evaluation of serum free amino acid concentration, has shown that threonine, glutamic acid, alanine, valine, isoleucine, leucine, γ-aminobutyric acid, and glycine have the highest diagnostic and prognostic value in dogs suffering from benign and malignant perianal tumours. Highly diagnostic and prognostic value in relation to benign tumour growth was also found while evaluating branched chain and acidic groups of amino acids. Serine, phenylalanine, lysine, and histidine, as well as alkaline amino acids were proved not to have diagnostic and prognostic value in dogs with the tumours. Thus, the evaluation of free amino acid concentration may serve diagnostic purposes and help in tumour malignancy differentiation. The elaborated experimental model may be used in further studies on neoplastic disease development and monitoring of applied treatment efficiency.

Sex-related differences of bone properties of pelvic limb and bone metabolism indices in 14-month-old ostriches (Struthio camelus)

ABSTRACT 1. Sex-related differences of long pelvic limb bones and serum bone metabolism indices were evaluated in 14-month-old female (N = 7) and male (N = 7) ostriches of similar body weights. 2. Densitometric parameters of femur, tibia and tarsometatarsus were determined using quantitative computed tomography (volumetric bone mineral density, calcium hydroxyapatite density and mean volumetric bone mineral density) and dual energy X-ray absorptiometry (bone mineral density and bone mineral content) methods. Geometrical parameters such as cortical bone area, cross-sectional area, second moment of inertia, mean relative wall thickness and cortical index were determined in the midshaft of bones. Mechanical properties of bones (maximum elastic strength and ultimate strength) were evaluated using three-point bending test. Serum concentrations of free amino acids, osteocalcin, N-terminal propeptide of type I procollagen, C-terminal telopeptides of type II collagen and total antioxidative capacity were also determined. 3. Bone weight and relative bone weight of all bones were significantly higher in males than in females. Significantly lower values of trabecular bone mineral density and calcium hydroxyapatite density were found in the trabecular bone of tibia in males. The highest number of the sex-related differences was observed in the tarsometatarsus where bone length, bone mineral content, cortical bone area, cross-sectional area and ultimate strength were higher in males. Serum concentrations of taurine, hydroxyproline, valine and isoleucine were significantly higher in males. 4. Higher loading of the tarsometatarsus in comparison to femur and tibia may be an important factor interacting with sex hormones in regulation of bone formation and mineralisation processes. Sex-related differences of bone properties were associated with increased serum concentration of selected amino acids in males.

Actylise treatment does not influence nitric oxide metabolites serum level

BACKGROUND Nitric oxide (NO) is synthesized by Nitric Oxide Synthases (NOS), the family of enzymes capable to conduct the conversion of Arginine (Arg) into the NO and Citrulline (Cit). Currently, only the administration of recombinant tissue plasminogen activator (rtPA) is recommended for acute ischemic stroke (AIS) treatment. To allow solubility of rtPA, Arg is added as a constituent of the drug. Our purpose was to check the effect of alteplase administration on NO metabolites concentration in the blood. METHODS Eighteen AIS patients were selected into the study. Nine of them received thrombolytic therapy (rtPA group). The serum samples were obtained at 3 time-points for rtPA group (time-point 0: 1st-4th hour of stroke; time-point 1: immediately after rtPA administration; time-point 3: on day 5-7 from stroke onset). Remaining patients (non-rtPA group) had blood collection at two time-points: time-point 1: 1st-10th hour of stroke and time-point 2: on day 5-7 of stroke. Arg and Cit were determined by the automated ion-exchange chromatography using Amino Acids Analyzer. NO serum level was indirectly evaluated with the usage of commercially available kits that measuring the nitrate/nitrite level. RESULTS Significant increase of Arg serum level was noticed at time-point 1, directly after the iv thrombolysis in comparison to non-rtPA group. However, the products of the reaction catalyzed by NOS (NO and Cit) did not rise after the thrombolysis. CONCLUSIONS Current study showed that Arg administration simultaneously with rtPA, as a constituent of Actylise, does not affect serum NO metabolites level.