Maria Laszczyńska

Membrane stability and mitochondrial activity of human-ejaculated spermatozoa during in vitro experimental infection with Escherichia coli, Staphylococcus haemolyticus and Bacteroides ureolyticus

The aim of the study was to examine an in vitro effect of the three bacterial strains (Escherichia coli, Staphylococcus haemolyticus and Bacteroides ureolyticus) on ejaculated spermatozoa with reference to sperm membrane integrity and mitochondrial activity. The study was carried out on swim‐up‐separated spermatozoa from 12 normozoospermic volunteers. Sperm plasma membrane stability was evaluated by the LIVE/DEAD Sperm Viability Kit and by the merocyanine 540 test. Mitochondrial activity was evaluated using the JC‐1 test as well as the NADH‐dependent NBT assay. The percentage of dead cells was significantly higher in spermatozoa treated with B. ureolyticus as compared to that of control spermatozoa (P < 0.01). All the bacterial strains applied affected sperm plasma membrane architecture measured by M540 test (P < 0.01). Moreover, the presence of E. coli or B. ureolyticus was connected with significant decrease in both the number of cells with high mitochondrial transmembrane potential (ΔΨm) and the cells with normal oxidoreductive function of mitochondria (P < 0.05 as compared to untreated cells). To conclude, the contact of bacteria with ejaculated spermatozoa can be a reason for severe injury of sperm membrane stability and mitochondrial activity with potential consequences for male fertility.

Relationship between the concentrations of heavy metals and bioelements in aging men with metabolic syndrome.

Heavy metals may exacerbate metabolic syndrome (MS) but abnormal serum concentrations of bioelements may also co-exist with MS. The primary aim of the study was to assess the relationship of blood heavy metal and bioelement concentrations and MS, in men aged 50–75 years. Heavy metals—lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As), tungsten (W), Macroelements—magnesium (Mg) and calcium (Ca), and microelements—iron (Fe), zinc (Zn) copper (Cu), chromium (Cr), molybdenum (Mo), selenium (Se) and manganese (Mn), body mass index (BMI), waist to hip ratio (WHR), abdominal circumference (AC) and blood pressure (BP), total cholesterol (TCh), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG), fasting plasma glucose (FPG), insulin, and Homeostasis Model Assessment—Insulin resistance (HOMA-IR). The men with MS showed statistically significant higher Zn and lower Mg concentrations. Those with diabetes had higher Ca concentration and lower Mg concentration. Cr and Mn concentrations were significantly higher in obese men. The participants with hypertension had lower Mg concentration. We found statistically significant positive correlations (W-TCh, W-LDL, Mg-TCh, Mg-LDL, Ca-TCh, Ca-LDL, Ca-insulin, Ca-HOMAR-IR, Zn-TG, Zn-insulin, Zn-HOMA-IR, Cu-BP systolic, Mn-BMI, Mn-AC, Mn-WHR, Mn-insulin, Mn-HOMA-IR, Se-TCh, Se-LDL, Se-TG, Se-insulin, Se-HOMA-IR, Cr-TCh, Cr-HDL, Cr-LDL, Cr-TG) and negative correlations (Cd-insulin, Hg-WHR, W-insulin, W-HOMA-IR, Mg-BMI, Mg-AC, Mg-WHR, Mg-BP systolic, Mo-insulin, Mn-HDL). Tungsten may contribute to lipid disorders. Magnesium appears to play the protective role in the occurrence of metabolic disorders. Microelements Mn, Cr and Se may intensify MS.

Serum Adiponectin and Leptin Concentrations in Relation to Body Fat Distribution, Hematological Indices and Lipid Profile in Humans

The purpose of the study was to evaluate the relationship between serum adiponectin and leptin concentrations and body composition, hematological indices and lipid profile parameters in adults. The study involved 95 volunteers (BMI from 23.3 to 53 kg/m2). Anthropometric parameters were measured: body weight and height, waist and hip circumference, waist-to-hip ratio, body fat mass (BMF), subcutaneous and visceral fat mass (SFM, VFM), lean body mass (LBM), skeletal muscle mass (SMM). In serum we determined adiponectin and leptin concentrations, extracellular hemoglobin, total bilirubin, as well as lipid metabolism (TCh, HDL-Ch, LDL-Ch, TG). Mean adipokine levels were significantly higher in women (p ≤ 0.01), adiponectin significantly negatively correlated with body height and weight, systolic blood pressure and absolute LBM and SMM values. The same relation was observed for erythroid system indicators and lipid indicators. A positive correlation was exceptionally found between adiponectin and HDL-Ch. LEP negatively correlated with some percentage rates (%LBM, %SMM). Only in women, we observed a positive correlation between LEP and body weight, BMI and WHR. Studies on ADPN and the ADPN/LEP ratio as a valuable complementary diagnostic element in the prediction and prevention of cardiovascular diseases need to be continued.

Morphology of ovaries in laron dwarf mice, with low circulating plasma levels of insulin-like growth factor-1 (IGF-1), and in bovine GH-transgenic mice, with high circulating plasma levels of IGF-1

BackgroundIt is well known that somatotrophic/insulin signaling affects lifespan in experimental animals, and one of the signs of aging is progressive gonadal dysfunction.MethodsTo study the effects of insulin-like growth factor-1 (IGF-1) plasma level on ovaries, we analyzed ovaries isolated from 2-year-old growth hormone receptor knockout (GHR-KO) Laron dwarf mice, with low circulating plasma levels of IGF-1, and 6-month-old bovine growth hormone transgenic (bGHTg) mice, with high circulating plasma levels of IGF-1. The ages of the Laron dwarf mutants employed in our studies were selected based on their overall survival (up to ~ 4 years for Laron dwarf mice and ~ 1 year for bGHTg mice).ResultsMorphological analysis of the ovaries of mice that reached ~50% of their maximal life span revealed a lower biological age for the ovaries isolated from 2-year-old Laron dwarf mice than their normal-lifespan wild type littermates. By contrast, the ovarian morphology of increased in size 6 month old bGHTg mice was generally normal.ConclusionOvaries isolated from 2-year-old Laron dwarf mice exhibit a lower biological age compared with ovaries from normal WT littermates at the same age. At the same time, no morphological features of accelerated aging were found in 0.5-year-old bGHTg mice compared with ovaries from normal the same age-matched WT littermates.

The effect of low and high plasma levels of insulin-like growth factor-1 (IGF-1) on the morphology of major organs: Studies of laron dwarf and bovine growth hormone transgenic (bGHTg) mice

It is well known that somatotrophic/insulin signaling affects lifespan in experimental animals. To study the effects of insulin-like growth factor-1 (IGF-1) plasma level on the morphology of major organs, we analyzed lung, heart, liver, kidney, bone marrow, and spleen isolated from 2-year-old growth hormone receptor knockout (GHR-KO) Laron dwarf mice (with low circulating plasma levels of IGF-1) and 6-month-old bovine growth hormone transgenic (bGHTg) mice (with high circulating plasma levels of IGF-1). The ages of the two mutant strains employed in our studies were selected based on their overall ~50% survival (Laron dwarf mice live up to ~4 years and bGHTg mice up to ~1 year). Morphological analysis of the organs of long-living 2-year-old Laron dwarf mice revealed a lower biological age for their organs compared with normal littermates, with more brown adipose tissue (BAT) surrounding the main body organs, lower levels of steatosis in liver, and a lower incidence of leukocyte infiltration in different organs. By contrast, the organs of 6-month-old, short-living bGHTg mice displayed several abnormalities in liver and kidney and a reduced content of BAT around vital organs.

Nuclear factor E2-related factor-2 (Nrf2) expression and regulation in male reproductive tract

BACKGROUND Nuclear factor E2-related factor-2 (Nrf2, Nfe2l2) plays an important, protective role in many tissues. However, information on molecular mechanisms of detoxification and drug metabolism regulated by Nrf2/NRF2 in testis and epididymis is scarce, but it may help to better characterize the function of blood-testis and epididymis barriers. METHODS Constitutive gene expression was analyzed by real time PCR with TaqMan Assay using ΔCT-method. Additionally, gene expression after treatment with oltipraz- specific Nrf2 inducer was evaluated using ΔΔCT-method. Cellular localization of the Nrf2 was visualized by immunohistochemical reaction. RESULTS The study showed that Nrf2 mRNA level in rat epididymis was higher than in testis. In human tissues, both testis and epididymis demonstrated similar expression levels of NRF2. Immunohistochemical analysis revealed NRF2/Nrf2 protein expression in testis and epididymis, which in the case of testis was dependant on spermatogenesis stage. Both in human and rat tissues constitutive expression of NQO1/Nqo1 was slightly higher in epididymis than in testis. Other Nrf2 regulated genes: GCLC/Gclc and UGT1A6/Ugt1a6 showed different ratios of testis/epididymis/liver expression levels. Treatment with oltipraz (Nrf2 inducer) resulted in significant induction of Nrf2 expression solely in corpus of epididymis. CONCLUSIONS Components of the Nrf2/NRF2 system along with coordinated genes are expressed in testis and epididymis. Moreover, some interspecies differences between rat and human were observed, which may impact extrapolation of experimental data into clinical findings. Studies on animal model showed that corpus of epididymis is the most responsive part of the male reproductive tract to oltipraz exposure at the gene expression level.

The effect of calorie restriction on the presence of apoptotic ovarian cells in normal wild type mice and low-plasma-IGF-1 Laron dwarf mice

BackgroundIt is known that caloric restriction extends lifespan and can minimize age-related dysfunction of the reproductive system. We became interested in how caloric restriction influences apoptosis, which is a crucial process that maintains ovarian cell homeostasis.MethodsWe examined ovarian cells in: 2.5-year-old wild type mice on caloric restriction (CR) or fed ad libitum (AL) and Laron dwarf mice (GHR-KO) at the same ages on CR or fed AL. Apoptosis was assessed by histochemical analysis on paraffin sections of ovarian tissue.ResultsMorphological and histochemical analysis revealed that CR improved reproductive potential in 2.5-year-old WT littermates and GHR-KO female mice, as indicated by the increased number of ovarian follicles. The level of apoptosis in ovarian tissue was higher in WT mice on a CR diet compared with WT mice on the AL diet. In GHR-KO mice, the level of apoptosis in ovaries was similar for mice on CR and on AL diets and bigger than in WT mice on CR.ConclusionsMorphological and histochemical analysis revealed a younger biological age of the ovaries in 2-year-old WT littermates and GHR-KO female mice on CR compared with animals fed AL.

Relationship between serum magnesium concentration and metabolic and hormonal disorders in middle-aged and older men.

The aim of this study was to determine the relationship between serum magnesium (Mg) concentrations in middle-aged and older men, and the occurrence of selected metabolic and hormonal disorders. The study involved a total of 313 men aged 50-75 years, and who underwent standard anthropometric measurements. Also determined were blood serum Mg, lipid parameters - total cholesterol (TCh), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG) and carbohydrate metabolism parameters - glucose, insulin (I). In men with a normal total testosterone (TT) concentration, serum Mg was significantly higher compared to patients with a total testosterone deficiency. The subjects with metabolic syndrome had lower serum Mg compared to patients without metabolic syndrome. Serum Mg concentration in type 2 diabetes mellitus was lower compared to men without diabetes. In patients with arterial hypertension, serum Mg concentration was lower than in patients without hypertension. Statistically significant negative correlations were found for the relationships between Mg concentration and body mass index, waist-to-hip ratio, abdominal circumference, arterial blood pressure. Statistically significant, positive correlations were found for the relationships between Mg concentration and TT, TCh, LDL concentrations. The lower serum Mg level may be conducive to the development of total testosterone deficiency, arterial hypertension, diabetes, and therefore metabolic syndrome.

Novel Morphological Findings of Human Sperm Removal by Leukocytes in In Vivo and In Vitro Conditions: Preliminary Study

Biological significance of increased number of leukocytes in ejaculate remains a subject of controversy. The aim of this research was to describe the morphological characteristics of the sperm elimination by leukocytes in in vivo and in vitro conditions using natural stimulator of the immune system—uropathogenic Escherichia coli, O75:HNT, isolated from semen.

Evaluation of spermatozoa of the rat in hyperprolactinaemia induced by Metoclopramide

Summary. Metoclopramide (MCP) in doses of 2.2 mg kg−1 body weight, being administered intraperitoneally to rats for 14 d, provokes a premature release of germ cells in the testes. The number of spermatozoa is reduced. In addition the spermatozoa exhibit defects of head and tail structures. After the administration of MCP for 14 d, followed by a 14‐d break in applying the drug, the number of spermatozoa in the lumen of the epididymal duct continues to be low. Many spermatozoa are damaged. Only after a 66‐d‐break in MCP administration is the morphology of the seminiferous epithelium restored, and a large number of spermatozoa appear in the lumen of the epididymal duct.