Sperm mitochondrial dysfunction and oxidative stress as possible reasons for isolated asthenozoospermia.

Reduced sperm motility, defined as asthenozoospermia, is a frequent cause of male infertility, and is mainly connected with the dysfunction of sperm mitochondria. The aim of this study was to identify the proteins, and thereby the metabolic pathways, responsible for asthenozoospermia, using 2-DE and MALDI-TOF MS, and correlate the results obtained with those of two mitochondrial tests: JC-1 and MitoSox Red. The JC-1 test was performed to test sperm mitochondrial activity, and the MitoSox Red test was performed to check whether the observed sperm poor motility is associated with mitochondrial reactive oxygen species (ROS) production. To identify proteins strictly connected with reduced sperm motility, men with isolated asthenozoospermia (n = 4 versus 10 normozoospermic controls) alone were included in the study. The proteomic analyses resulted in the identification of 25 sperm proteins that are differentially expressed in asthenozoospermic individuals. Most of the identified proteins were downregulated and were involved in energy production; however, we have also identified structural sperm proteins and proteins secreted by the epididymis. The latter, together with the results from MitoSox Red assay, may provide insights into the pathophysiological basis of asthenozoospermia.

Expression pattern of ISL-1, TTF-1 and PAX5 in olfactory neuroblastoma.

Olfactory neuroblastoma (ONB) is a rare neoplasm of the sinonasal area with neuroendocrine differentiation. ISL-1, TTF-1 and PAX5 are transcription factors that are frequently upregulated in tumors showing neuroendocrine differentiation. The aim of our study was to evaluate these markers in a group of ONBs. We included 11 ONBs from 4 large university hospitals. Immunohistochemical expression of TTF-1, PAX5 and ISL-1 was evaluated. TTF-1, ISL-1 and PAX5 were expressed in 3/11 cases (27.27%, h-score: 3-45), 7/11 cases (63.64%, h-score: 23-200), and in 3/11 cases (27.77%, h-score 3-85), respectively. The patient with the strongest PAX5 reactivity exhibited an aggressive clinical course with rapid dissemination to the spine and death shortly after the diagnosis. No significant correlation in the expression of PAX5 and TTF-1 ( = 0.43; p = 0.18) was observed. ISL-1 is widely expressed in tumors with neuroendocrine differentiation and therefore of limited value in their differential diagnosis. TTF-1 positivity does not exclude the diagnosis of primary ONB, although usually only a small percentage of cells are positive. PAX5 expression is infrequent (27.27%) in ONB; however, if present it can be associated with a very aggressive clinical course.

Changes in lipid metabolism during last month of pregnancy and first two months of lactation in primiparous cows - analysis of apolipoprotein expression pattern and changes in concentration of total cholesterol, HDL, LDL, triglycerides.

The final weeks of pregnancy and period of increasing lactation abound with adaptive changes in the intensity of metabolic processes. Maintaining the homeostasis of an organism in prepartum and postpartum periods is the key condition in maintaining the health of the mother and the fetus/calf. The aim of the study was to analyze physiological changes in lipid metabolism in cows during the last month of first pregnancy and in the first two months of lactation, based on the expression of identified apolipoproteins and changes in selected parameters of the lipid metabolism in peripheral blood plasma. Statistically significant changes in the expression of identified apolipoproteins were observed for apolipoprotein A-1 precursor, apolipoprotein A-IV precursor, apolipoprotein E precursor and apolipoprotein J precursor. The lowest expression of the apolipoproteins was noted around parturition and higher expression was observed during the final weeks of pregnancy and during lactation. Tendencies of changes in the concentration of total cholesterol, HDL and LDL were similar in blood plasma from analyzed cows - in the last month of pregnancy a decrease was observed and subsequently an increase in the first two months of lactation was noted. In contrast to abrupt changes observed for total cholesterol, HDL and LDL, changes in concentration of triglycerides were not that extensive and during lactation this parameter was rather stable. Evaluation of changes in the analyzed parameters may contribute to a better understanding of the changes in lipid metabolism occurring in the body of pregnant and lactating young cows.

Feeding milk replacer instead of whole milk affects blood plasma proteome and lipid profile in preruminant calves.

The study was undertaken to determine the effect of feeding milk or milk-replacer on the blood plasma proteome and lipid profile in calves during the second week of life. Feeding milk-replacer significantly decreased the expression of plasma apoA-I. Age of calves affected apoA-I expression, which was higher on the 8th than on the 11th and 14th day of life. A significant effect of interaction between diet and age was also observed. The expression of apoA-IV, was significantly affected by diet and was lower in calves fed milk replacer. Expression of this protein was significantly lower at the 8th day of life and was up-regulated in the calves fed milk-replacer at the second week of life. Calves fed milk-replacer had greater expression of haptoglobin, which differed significantly between days of blood sampling, being higher on the 8th than on the 11th and 14th day. The interactive effect of diet and age affected haptoglobin expression, which was successively down-regulated in calves fed milk re- placer. Diet had a significant effect on the plasma lipid profile. Animals fed milk had a greater concentration of TC, HDLC and LDLC. The composition of milk-replacer, especially fat source, is probably the main factor that affects expression of proteins involved in cholesterol metabolism and level of components of lipid profile in calves fed formula. We claim that the initially increased level of haptoglobin, followed by its decrease during the second week of life in calves fed milk-replacer may indicate the presence of short-term stress induced by changes in the feeding system.

Mapping of polar fox renal cortex proteins using two-dimensional gel electrophoresis and mass spectrometry--a preliminary study.

The aim of the present study was to establish protein map of polar fox (Alopex lagopus) renal cortex. Kidney cortex proteins of isoelectric point ranging from 3 to 10 were analysed using two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Sixteen protein spots corresponding to thirteen different gene products were identified. These proteins were divided into following groups: lipid and fatty acid metabolism, amino acid metabolism, energetic pathways, regulatory proteins, transport proteins and structural proteins. This is the first attempt to create reproducible 2-D map, of renal cortex proteins characteristic for polar foxes, used as animal model for carnivores. It is worth emphasizing that the results of this study may broaden currently available protein databases.

Blood plasma protein and lipid profile changes in calves during the first week of life.

The present study was undertaken to determine blood plasma protein and lipid profile changes in healthy Polish Holstein-Fresian calves of Black-and-White variety. Blood was drawn immediately after birth, before first colostrum intake and at the 3rd, 6th, 12th, 24th, 36th, 48th and 72nd hour of life. Subsequent four blood samples were collected at 24 hour intervals until the 7th day of life. Plasma proteins within the isoelectric point ranging from 3.0 to 10.0 were separated using high resolution two-dimensional electrophoresis. Among the 74 protein spots detected and analyzed, 16 were significantly altered during the first week of life. Differentially expressed spots were excised from the gels and subjected to peptide mass fingerprinting using MALDI-TOF MS. In total, 12 spots were successfully identified, which correspond to three proteins, namely: apolipoprotein A-I, apolipoprotein A-IV and fibrinogen gamma-B chain. A gradual increase in plasma triglyceride, total cholesterol, HDL and LDL cholesterol values was shown during the first seven days of calves life. The lowest concentration of these indicators were observed at birth and was followed by a rapid increase during the first week of postnatal life. These changes appear to be related to the transition in energy sources, from a maternal nutrient supply comprising mainly carbohydrates and amino acids to a diet which was rich in fat--colostrum and milk. This was reflected by the intense up-regulation of plasma proteins related with lipid transport and lipoprotein metabolism during the first week of life.

Defining the blood plasma protein repertoire of seven day old dairy calves - a preliminary study.

During the early postnatal period in calves various adaptational changes occur. These functional, morphological and also metabolic alteration are reflected by blood plasma protein changes as they are secreted and shed from many cells and tissues. Blood plasma protein pattern of an adult cattle differs in some respect when compared with neonatal calves. There exist a very few data concerning 2-D maps of neonatal calves blood plasma. The above prompted us to establish protein pattern of this biological fluid characteristic of healthy, 7 day old, Polish Black-and-White (Polish Friesian) breed calves. Blood plasma proteins of the isoelectric point ranging from 4.0 to 7.0 were analyzed by the aid of high resolution two-dimensional electrophoresis (2-DE). Subsequently, 79 excised protein spots corresponding to 23 different gene products were identified using matrix-assisted laser desorption/ionisation mass spectrometer (MALDI-TOF MS). Protein map obtained in the present study may be useful in assessing the changes in the calves blood plasma protein profiles occurring in response to different physiological and/or pathophysiological factors.

Urinary excretion of low molecular weight proteins in goats during the neonatal period.

Urinary protein excretion occurs in neonates of many animal species, as well as in human neonates. However, the incidence, dynamics, and mechanism of proteinuria have not been unambiguously explained. The aims of this study were to investigate into excretion of selected protein fractions of molecular weight less than 69 kDa (LMW), evaluation of intensity and dynamics of changes during the first month of kids' life, and an attempt to explain the causes of neonatal proteinuria. The analysis were carried out on 16 kids of White Improved goats, over the period from birth until 30 days of age, using clearance methods. Urine proteins were separated electrophoretically (SDSPAGE), and their concentration and percentage content was determined by densitometric method with the use of archiving and image analysis software. The proteins found in the urine were grouped as HMW, LMW and albumin. For six fractions of LMW proteins, excretion rates and percentage content of the urinary total LMW protein pool were calculated. It has been demonstrated that neonatal proteinuria in goat kids is associated with a high level of excretion of proteins of lower molecular weight than albumin (69 kDa). A strong dynamics of changes in excretion of particular LMW protein fractions with age was observed, which may imply not only an increased permeability of glomerular filtration barrier, especially over the first days of life, but also a selectivity of reabsorption mechanisms in the nephrons. An increased permeability of glomerular filtration barrier for proteins during the first days of life may represent the adaptive mechanism for removal of protein excess from the organism. The urinary LMW protein pool may also contain proteins resulting from the hydrolysis in the tubular cells.

Plasma proteome analysis: 2D gels and chips.

The knowledge of concentration, modification and interaction of proteins is fundamental in determining the phenotype of living organisms. Plasma, the primary clinical specimen, contains numerous and diverse proteins. The functions of these proteins are as manifold as the diversity of the protein themselves. Many of them have been largely used for many years as biomarkers of diseases and indicators of the physiological functions. The study of plasma proteome promises to be a significant advance in various areas of biological and clinical research. Two-dimensional polyacrylamide gel electrophoresis is considered as a primary tool in separating thousand of plasma proteins. This approach enables comparing normal and diseased samples revealing differently expressed proteins. Other proteomic techniques suitable for plasma analysis such as protein microarrays are now either established or are still being improved. This article briefly reviews the application of two-dimensional electrophoresis and the current status of technical aspects for plasma proteome.

[Tuberculous tubo-ovarian abscess as a distant complication of Caesarean section]. / Gruzlicze zapalenie przydatków jako odlegle powiklanie po cieciu cesarskim.

A 37-year-old patient, two mounts after the third caesarean sections, was rehospitalised due to severe lower abdomen pains and fever. She had been treated with antibiotics several times with no apparent result. Laparotomy was performed and the tubo-ovarian abscess was resected. The histological diagnosis was--adnexal tuberculosis. The postoperative period was febrile with prolonged inguinal lymphatic nodes reaction. She was constantly treated with broad-spectrum antibiotics and left hospital after 30 postoperative days. The disease was probably the result of symptomless lung tuberculosis, she had suffered from in the past.

[Microbiological evaluation of the effectiveness of gynalgin in the treatment of vaginitis]. / Ocena mikrobiologiczna skutecznosci Gynalginu w zapaleniach pochwy u kobiet.

The microbiological effectiveness of the preparation Gynalgin produced by POLFA Pharmaceutical Works in Rzeszów was assessed in cases of vulvovaginitis in 55 patients with clinically diagnosed inflammatory conditions of the lower genital tract, who were given Gynalgin tablets in 10-day courses. Vaginal smears were examined three times for the presence of bacteria, fungi and trichomonas vaginalis (before and immediately after the treatment, and two weeks later). In the initial examination in five vaginal smears mixed bacterial flora was found, in 6 smears trichomonas was present, in 4--bacteria and fungi, and in one--trichomonas and fungi. After the treatment in control examinations I and II the number of the isolated bacterial strains was lower, trichomonas was no longer present, and the number of fungi was reduced evidently. In the light of these microbiological examinations Gynalgin was found to exert a strong fungicidal, bactericidal and antitrichomonal activity, and the results of laboratory investigations agreed with those of clinical trials of Gynalgin effectiveness.

Seasonal variations of antipyrine pharmacokinetics parameters in calves.

In the experiment the values of parameters of antipyrine kinetics were defined (Vd - volume of distribution, t0.5 - half-life, CA - metabolic clearance) in calves in January, April, July and October 1986 and in January and July 1987. Statistically significant increase of distribution volume and increase of hepatic antipyrine clearance were recorded, as well as significant shortening of half-life of this substance in the organism of tested animals in summer 1986 and 1987, compared with the remaining seasons of the year. Antipyrine pharmacokinetics in winter, spring, autumn 1986 and winter 1986/87 did not differ significantly.

Pharmacokinetics of antipyrine in calves during first 35 days of life.

The experiment was carried out on 10 bull-calves of black-white breed, aged 2-35 days. The following parameters of antipyrine pharmacokinetics were determined: Vd - volume of distribution, t0.5 - biological half-life, CA - metabolic clearance. It has been shown statistically that the volume of antipyrine distribution (expressed in 1/kg) was significantly decreasing with age of examined animals. The antipyrine half-life proved to be the longest on the 20th day of calves' life and the shortest on the 10th and 30th day. The maximum values of metabolic clearance of antipyrine were observed in calves aged 10 days and the minimum ones in animals aged 20 days. In the cases of t0.5 and CA a tendency to changes of values of these parameters of antipyrine pharmacokinetics in 10 days periods was observed.

Monthly variations in the pharmacokinetics of antipyrine in calves.

In the experiment (on the basis of values of antipyrine (phenazone) pharmacokinetics parameter), biotransformation activity of calves liver during a year was determined. The experiment was carried out on calves aged 28-30 days. Volume of distribution (Vd), half-life (t0.5) and metabolic clearance (CA) of antipyrine were from month to month determined. Not significant changes between values of Vd, t0.5 and CA in several months were observed. Results of experiment indicated that calves' liver is characterized by the relatively stabile biotransformational activity during the whole year.

[The influence of ACTH and hydrocortisone on drug kinetics in calves in the example of the antipyrine test]. / Wplyw ACTH i hydrokortyzonu na kinetyke leku u cielat na przykladzie testu antypirynowego.

The aim of the experiment carried out on 2 groups (to comprise 8 specimens) bull calves, of ncb. breed, at the age of 60 to 65 days and of an average body weight 90 +/- 5 kg, was to define the effect of intramuscular ACTH (0.2 j.m./kg b.w.) and hydrocortisone (0.2 mg/kg b.w.) injections on the hepatic biotransformation of antipyrine. No changes were noticed in the quantity of antipyrine distribution space Vd under the influence of the hormonal preparations applied. Shorter time of antipyrine halflife from the body T/2 was observed, yet the differences stated have not confirmed statistically. ACTH and hydrocortisone injections caused the increase in constant elimination value of antipyrine from the organism k as well as the increase in the quantity of hepatic antipyrine clearance ClA. The differences observed have not turned out to be statistically significant.

[Sodium, potassium and chloride levels in the kidney tissue of piglets in the postnatal period]. / Poziom elektrolitów: sodu, potasu i chlorków w tkance nerkowej prosiat w okresie postnatalnym.

In this paper the level of Na, K and Cl was determined on 108 kidney slices obtained from 12 kidney, removed from 6 clinical healthy piglets. The animals were 4 weeks old. The kidney tissue prepared by method as described by Appelboom et al. (1958). The electrolyte concentration in the cortex, outer medulla and inner medulla was calculated per 1 kg wet and dry tissue. There is a distinct (significant differences) electrolyte gradient in the piglet kidney tissue (the maximum electrolyte concentration in the papilla area). Low Na content even in inner medulla, indicates little capacity for urine concentration in the 4 week old piglets. Individual differences of the electrolyte content in kidney tissue, may indicate different rate of functional development of piglets kidney. No significant difference between left and right kidney was stated in the electrolytes concentration.

[Effect of hypophysin and desoxycorticosterone acetate on kidney function in 3-month-old calves]. / Wplyw hipofizyny i octanu dezoksykortykosteronu na czynnosc nerek u trzymiesiecznych cielat.

The experiment has been carried out on 6 bulls of cb. breed at the age of 3 months. Clearance methods were used to examine the hypophysin (0.1 u V/kg) and desoxycorticosterone acetate (0.1 mg/kg) influence on the size of diuresis, glomerular filtration and excretion of the following: sodium, potassium, chlorides, calcium, magnesium and inorganic phosphorus with urine. Three months old calves had (in comparing to calves in neonatal period) higher glomerular filtration level and lower coefficient of endogenous creatinine purification, however the GFR did not still reach the GFR level of adult animals. Hypophysin and desoxycorticosterone acetate influenced both the process of glomerular filtration (decrease) and tubular transportation of water, Na, K and Cl. After the acetate had been applied, higher condensation of urine was noticed than after the application of hypophysin. Administering DOC to the calves caused decrease in sodium and chlorides excretion with urine as well as increase in kidney purification of potassium. Increased excretion of potassium ion and the decrease in urine pH after DOC injection may suggest that older calves compared with the ones at the age of 2-5 weeks have their kidney mechanism regulating potassium and acid-basic balance developed and efficient. Significant effect of hypophysin and desoxycorticosterone acetate on the process of calcium, magnesium and inorganic phosphorus excretion with the urine has not been stated.

[Kidney function in calves in the neonatal period. I. Studies of the capacity to store electrolytes Na, K, Cl, Ca, Mg and Pn]. / Funkcja nerek u cielat w okresie neonatalnym. I. Badania nad zdolnoscia do oszczedzania elektrolitów Na, K, Cl, Ca, Mg i Pn.

The capability of the kidneys of 12 bull-calves at the age of 7-35 days of life to save electrolytes Na, K, Cl, Ca, Mg, Pn were examined. The examinations were made with the clearance methods using inulin as the indicator of the glomerular filtration of plasma. It was proved, that as the calves grow and develop, there arise changes in the glomerular filtration, tubular resorption and in the excretion of the examined minerals in urine. There was observed, at the same time, the relatively stabile level of these electrolytes in blood. It shows the functional development of the calves kidneys and the fact, that in the proper environmental conditions and particularly at the proper feeding, the kidneys are able to maintain the favorable, from the point of view of the internal balance and the needs of the developing organism, level of the electrolytes.

[Kidney function in calves in the neonatal period. II. Studies of the capacity to reduce and increase urine concentration]. / Funkcja nerek u cielat w okresie neonatalnym. II. Badania nad zdolnoscia do rozcienczania i zageszczania moczu.

Examinations were made with the use of same materials as in Part I and according to the same methodology. The great capability of the calves kidneys to economise water was observed. The kidneys are able already in the second week of calves life to reduce and increase urine concentration within limits from 65.70 to 955.54 mmol/kg H2O, keeping at the same time molality of the body fluids on relatively the same level. It may prove high efficiency of the hypothalamic pituitary antidiuretic system. It seems, that because of the probable immaturity of tubules it was difficult to create the high osmotic gradient in the renal medulla what is confirmed by the changing clearances of the free water and the resorption water. It also seems, that the rate with which urine is passing through the renal tubules is of great importance in the process of thickening the urine in this period.