Dietary cholesterol and oxidised cholesterol: effects on sperm characteristics, antioxidant status and hormonal profile in rats.

Present study was designed to compare the potential effects of high serum levels of LDL and oxidised LDL (OxLDL) on spermatogenesis parameters in male Wistar rats. Animals were allocated into three groups and were fed for 14 weeks with normal, cholesterol-rich and oxidised cholesterol-rich diets. Blood lipid profile, sex hormones level, as well as sex organs weight were evaluated. The sex organs weight in oxidised cholesterol-fed group was significantly reduced (P < 0.05). Spermatozoa count in the group with high serum concentration of OxLDL (64 ± 4.2 × 10(6) ) was markedly lower (P < 0.01) than that of normal rats (87 ± 4.1 × 10(6) ) and rats with high serum level of LDL (90 ± 6.3 × 10(6) ). Similarly, the percentage of viable spermatozoa was significantly (P < 0.001) decreased from 78% to 52% by high level of OxLDL in serum. While, nonoxidised LDL did not have suppressive effects on spermatogenesis and organs weight. Consistent with these effects, the serum concentration of sex hormones including FSH (P < 0.001), LH (P < 0.001) and testosterone (P < 0.01) was significantly decreased only in rats with high level of OxLDL but not in rats with high level of nonoxidised LDL. In conclusion, high OxLDL level showed higher destructive effect on reproductive system compared to the high LDL level.

Fluorescence emission quenching of RdB fluorophores in attendance of various blood type RBCs based on Stern-Volmer formalism.

In this work, the optical properties of Rhodamine B (RdB) are investigated in the attendance of various red blood cells (RBCs). RdB fluorophores, as biological markers, is excited using SHG-CW Nd:YAG laser at 532 nm. In fact, the addition of biomolecules of interest to the reference fluorophore notably changes the fluorescence properties of the suspension. Here, laser induced fluorescence (LIF) spectrophotometry based on Stern-Volmer quenching formalism and field emission scanning electron microscope (FESEM) are employed here. According to the given fluorescence spectra, the spectral shift of emissions as well as quenching coefficients are assessed subsequently. The Stern-Volmer formalism is used to determine the quenching coefficients. In fact, RdB + RBCs suspensions contain a plenty of bioconjugates leading to the signal reduction and notable red shift in RdB fluorescence emissions. Furthermore, it is demonstrated that the positive blood type RBCs exhibit the higher quenching coefficients and the larger red shifts against those of negative blood types. This mainly arises from the nature of specific sugar antigens available on the RBC membranes as to N-acetylgalactosamine and galactose attached to the O-antigen terminal would enhance further quenching of the species. Moreover, a significant correlation appears between Stern-Volmer coefficients and the corresponding RBCs. In fact, distinct discrepancy takes place in quenching coefficients in terms various positive/negative blood types to envisage a facile method of blood typing.

Safe short circuit in cataract surgery: Incidence and risk factors for intraoperative medical action.

Cataract surgery has become the most frequent surgical procedure performed every year in Western countries. Perioperative patient circuit has to be adapted to the important medical needs and progress. Hence, a secure short circuit (SSC) for surgeries of the anterior segment of the eye under topical anaesthesia was created. Patients included in the circuit are selected first by surgeons and answer a medical questionnaire, they do not have any preoperative evaluation by anaesthesiologist, are monitored during surgery by the surgical team and in case of problem an intraoperative medical action (IMA) can be performed. We conducted a retrospective observational incidence study of the occurrence of the IMA, followed by a case control study. The primary outcome was to identify risk factors of IMA among the patients' medical history. Out of 2744 screened patients, 1592 patients were included during the period of November 2015 to November 2017. The rate of IMA was 5%, 81% of them presenting with intraoperative high blood pressure (HBP). In the case control study part, stepwise regression analysis revealed that a history of HBP and insulin-dependent diabetes (IDD) was significantly correlated with IMA (respectively, adjusted odds ratio 1.7, P=0.005 and 2.6, P=0.002). The low incidence of IMA showed that the SSC is a safe tool thanks to a selection and an optimised and secure pathway. A history of HBP and IDD was significantly associated with the occurrence of IMA. Therefore, an optimisation of the perioperative period would be beneficial in these cases.

The Effect of Prednisolone on miR 15a and miR16-1 Expression Levels and Apoptosis in Acute Lymphoblastic Leukemia Cell Line: CCRF-CEM.

BACKGROUND: Glucocorticoids are probably the most important drugs in the treatment of childhood acute lymphoblastic leukemia (ALL). Prednisolone exerts its effect by induce apoptosis in lymphoid lineage cells. Micro RNAs are 18-24 nucleotides RNA implicated in the control of essential biological functions, including apoptosis. In the following study, the effect of prednisolone on the expression of miR 15a & miR16-1 and apoptosis in the CCRF-CEM cell line is investigated. METHODS: The cell line of CCRF-CEM was cultured in standard conditions. The changes in the miR 15a and miR 16-1 expression levels were determined by Real Time-PCR technique. Also, the apoptosis is evaluated by flow cytometry using Annexin V and PI staining. RESULTS: This study revealed that, the prednisolone induced apoptosis in a time dependent manner. Prednisolone in concentration of 700 µM was significantly increased the expression of miR 16-1 and miR 15a after 24 h and 48 h treatment (p<0.05). CONCLUSION: prednisolone-induced apoptosis might be mediated by up-regulation of these 2 miRNAs in CCRF-CEM cells.

Investigation of the memory impairment in rats fed with oxidized-cholesterol-rich diet employing passive avoidance test.

BACKGROUND: Recent studies have shown that hypercholesterolemia, besides being a risk factor for cardiovascular diseases, has also toxic effects on central nervous system. The design of the present study was to investigate the effects of dietary cholesterol and oxidized cholesterol on cognitive function. METHODS: Male Wistar rats were randomly divided into 3 groups. The animals were fed with three normal, 2% cholesterol-rich, and 2% oxidized cholesterol-rich diets for 14 weeks. Memory impairment was analyzed by passive avoidance test. Coenzyme Q10 content was also measured by a validate RP-HPLC method. Besides, lipid peroxidation in serum and brain tissue was determined by malondialdehyde concentration measurement. RESULTS: The results showed that feeding rats with high oxidized cholesterol diet for 14 weeks significantly impaired the cognitive function compared to the normal (P<0.001) and high cholesterol-fed groups (P<0.01). The memory impairment was positively correlated to the serum level of the oxidized LDL; it was significantly associated with the increased malondialdehyde concentration on the brain tissue of both groups (P<0.05 and P<0.001, respectively). The total antioxidant level in the serum was also decreased in rats fed with the oxidized cholesterol (P<0.05). Moreover, the brain coenzyme Q10 content was significantly declined in the animals fed with the oxidized cholesterol-rich diet compared to the animals fed with the normal (P<0.01) and cholesterol-rich diets (P<0.05). CONCLUSION: The results suggested that the high dietary intake of the oxidized-cholesterol might impair the memory that could be correlated to the oxidative stress and declined the coenzyme Q10 content of the brain tissue.

Fibronectin: source of mannose in a highly purified respiratory mucin.

Human bronchial mucin, solubilized in an aqueous solution of sodium azide and protease inhibitors, was purified by molecular sieve chromatography. The mucin was purified by Sepharose 4B and 2B column chromatography. Chemical analyses of this preparation showed a typical mucin profile of amino acids and carbohydrates, except for the presence of an appreciable amount of mannose. Sodium dodecyl sulfate-polyacrylamide gel (5%) electrophoresis of this material showed a high Mr glycoprotein at the top of the gel and two additional bands with mobilities of fibronectin subunits (230 and 210 kD). The fibronectin was separated from the mucin by geletin-Sepharose column chromatography, and the fibronectin eluted from the column was immunologically similar to fibronectin purified from human serum. Ion-exchange chromatography of purified mucin resulted in neutral and acidic fractions. The neutral mucin was the major component. Chemical composition of these two fractions indicated that the amount of threonine, serine, and sialic acid was higher in the acidic fraction, whereas the neutral fraction contained more proline, aspartic acid, leucine, glycine, fucose, and galactose than the acidic fraction.

Neutral and acidic human tracheobronchial mucin. Isolation and characterization of core protein.

Human bronchial mucin from a patient suffering from chronic bronchitis was solubilized in aqueous solution containing sodium azide and protease inhibitors and purified by Sepharose 4B and 2B column chromatography. The mucin was further purified by cesium bromide density gradient centrifugation. Sodium dodecyl sulfate-polyacrylamide gel (7.5%) electrophoresis of this material showed high-molecular-weight mucin component(s) at the top of the gel. Chemical analysis of this preparation indicated a typical mucin profile of amino acids and carbohydrates. Ion-exchange chromatography resulted in resolution of the purified mucin into neutral and acidic fractions. Comparison of the chemical composition of these two fractions showed higher mole percentage of threonine, serine, sialic acid, and sulfate in the acidic fraction. Chemical deglycosylation of the purified mucin preparation with trifluoromethane sulfonic acid was carried out at 20 degrees C for 3 1/2 h. Sialic acid, fucose, galactose, and N-acetylglucosamine were completely removed, whereas traces of N-acetylgalactosamine were still detected. High-pressure liquid chromatography of the deglycosylated products from native, neutral, and acidic mucin preparations resulted in a principal peptide, P1, with identical amino acid composition. Cyanogen bromide (CNBr) treatment of the peptide P1 from neutral and acidic mucins and subsequent fractionation of the fragments by high-pressure liquid chromatography resulted in similar peptide profiles. The P1 peptide fraction was further subjected to high-pressure liquid chromatography in a second solvent system, which resulted in two peaks, P1a and P1b. Gel filtration of both peptides in 6 M guanidine hydrochloride indicated a single peak with molecular weight of approximately 97 kDa. The amino acid profile of the two peptides was dominated by high levels of threonine, serine, and proline, which combined accounted for nearly 39% of the total residues, and in most respects, the profile resembled that of native mucin. End-group analysis of the peptide P1a indicated a blocked N-terminus, whereas serine was found to be the N-terminal amino acid in the peptide P1b. Rabbit antibodies prepared against the peptide P1 from native tracheal mucin reacted strongly with neutral and acidic mucin as well as the mucin from human colon. Both neutral and acidic human tracheal mucins were immunologically reactive with mouse monoclonal antibody HMPFG-2, which was prepared against human mammary mucin. However, the response of this antibody to human colonic mucin was rather weak.

CoQ10 and L-carnitine attenuate the effect of high LDL and oxidized LDL on spermatogenesis in male rats.

BACKGROUND: It is estimated that one-third of infertility cases are due to male factors. Hyper-cholesterolemia is a social problem in many developed countries and contributed with a heterogeneous group of disorders characterized by an excess of cholesterol and its derivatives in the blood stream. PURPOSE: The objective of the present study was to investigate the protective effects of coenzyme Q10 and L-Carnitine supplementation on semen parameters, sperm function and reproductive hormone profiles in male Wistar rats with high LDL and Oxidized LDL (OxLDL) blood levels. METHODS: Animals were fed with cholesterol and oxidized cholesterol-rich diets for 14 weeks to elevate the LDL and OxLDL blood level, respectively. Pretreatment with coenzyme Q10 (10 mg/kg/day, oral) and L-Carnitine (350 mg/kg/day, oral) were conducted for 5 consecutive weeks. Sex hormones levels, malondialdehyde and total antioxidant concentrations, as well as testis, epididymis and seminal vesicle weight were also analyzed. RESULTS: Following high LDL and OxLDL blood levels, decrease in the sperms count and viability, weights of testis, epididymis and seminal vesicle as well as concentration of testosterone and LH hormone were observed. On the other hand, in contrast to reduction of total antioxidant level, malondialdehyde concentration, both in serum and testis, was increased. However, pretreatment with L-carnitine and coenzyme Q10 increased serum sex hormones level and improved semen parameters significantly. CONCLUSION: Overall, pretreatment with coenzyme Q10 and L-Carnitine attenuated the destructive effects of high LDL and oxidized LDL levels on spermatogenesis parameters in male rats.

Reduction of coenzyme q10 content: a possible effect of isoproterenol on heart failure and myocardial infarction in rat.

Myocardial infarction (MI) was induced by subcutaneous injection of isoproterenol (ISO) to investigate the effect of ISO on Coenzyme Q10 (CoQ10) content of myocardium and subsequent effects on lipid peroxidation, electrocardiogram pattern and hemodynamic parameters of the rat's heart.36 male Wistar rats were divided randomly into 6 groups. To induce heart failure (HF) and MI, 10 and 100 mg/kg of ISO was administered subcutaneously for 10 and 2 consecutive days, respectively. The effects of ISO on myocardium CoQ10 content, concentration of malondialdehyde, ECG pattern and hemodynamic parameters of heart were analyzed.ISO-treated rats showed significant alteration in heart hemodynamic parameters such as reduction of left-ventricular systolic pressure, maximum and minimum rate of developed left ventricular pressure, besides increase of left ventricular end-diastolic pressure. Significant depletion of heart CoQ10 content (from 4.57 and 4.55 µg/100 mg tissue in control groups to 2.85 and 2.89 µg/100 mg tissue in ISO-induced HF and MI groups respectively) and increase in tissue levels of malondialdehyde (47.1 and 53.8 nmol/100 mg tissue in ISO-induced HF and MI groups, respectively) were also observed in ISO-treated animals compared with the normal animals (17.4 and 18.8 nmol/100 mg tissue in control groups, respectively). Additionally CoQ10 improved ISO effects on hemodynamic parameters and ECG pattern in ISO-induced HF and myocardial injury.The present findings have demonstrated that the cardiotoxic effects of ISO such as oxidative damage and hemodynamic declination might be related to depletion of CoQ10 concentration.

Effects of L-carnitine and coenzyme q10 on impaired spermatogenesis caused by isoproterenol in male rats.

Nowadays, cardiovascular diseases and male infertility are two big health problems in industrial countries.The aim of the present study was to investigate the protective role of coenzyme Q10 and L-Carnitine pretreatment in the impaired spermatogenesis caused by isoproterenol (ISO) in male rats.Thirty-two male Wistar rats were allocated in 4 groups. ISO was injected for 2 consecutive days (100 mg/kg) in ISO treated groups. Before ISO administration, pretreatment with Coenzyme Q10 (10 mg/kg/day) and L-Carnitine (350 mg/kg/day) were conducted for 20 consecutive days. Sex hormones level, malondialdehyde (MDA) and total antioxidant concentration as well as testis, epididymis and seminal vesicle weight were investigated.Increase in the concentration of MDA and decrease in total antioxidant level was observed following ISO administration. Accordingly, the sperm viability as well as testis, epididymis and seminal vesicle weights were decreased. In the case of sex hormones, the testosterone and LH levels were decreased and the concentration of FSH was increased. Pretreatment with L-carnitine and Coenzyme Q10 significantly decreased the MDA level and increased total antioxidant, LH and testosterone levels. Pretreatment with L-carnitine and Coenzyme Q10 also improved semen parameters and organs weight which were impaired by ISO administration.L-carnitine and Coenzyme Q10 pretreatment could protect spermatogenesis in male rats with ISO administration.

Atorvastatin reduces the myocardial content of coenzyme Q10 in isoproterenol-induced heart failure in rats.

The present study was aimed to study the effects of different doses of atorvastatin on Co Q10 content in the myocardium tissue in rats. A subcutaneous injection of isoproterenol (5 mg/kg/day) for 10 days was used for the induction of heart failure. Rats were randomly assigned to control, treatment with atorvastatin (5, 10, 20 mg/kg/day) and treatment with atorvastatin plus coenzyme Q10 (10 mg/kg/day). Coenzyme Q10 content of myocardium was measured using HPLC method with UV detector after hemodynamic parameters measurements. The malondialdehyde (MDA) content of the myocardium was evaluated in order to determine coenzyme Q10 antioxidative effect. A high dose of atorvastatin (20 mg/kg/day) was significantly reduced the myocardium content of coenzyme Q10 as compared with isoproterenol treated group (p<0.001). Compared with atorvastatin alone treated animals, co-administration of coenzyme Q10 with atorvastatin was improved the level of coenzyme Q10 in the myocardium (p<0.05, p<0.001). Increasing the dose of atorvastatin also led to increase in MDA content of the myocardium (p<0.01). Serum lipid profile showed no changes in atorvastatin treated groups. The results of this study demonstrate that high doses of atorvastatin reduce coenzyme Q10 content of the myocardium and increase lipid peroxidation in myocardium which is reversed by coenzyme Q10 co-administration.

Effects of LDL and oxidized LDL on cardiac function in isoproterenol-induced myocardial infarction in rat.

Notable discussions have been developed over the distinctive effects of LDL and oxidized LDL (OxLDL) on myocardial functions. The aim of the present study was to investigate the effects of OxLDL on electrocardiogram and hemodynamic parameters of rat's heart in isoproterenol (ISO)-induced myocardial infarction (MI) model.Male Wistar rats were allocated in to 6 groups and receive one of the 3 formulated diets (standard, cholesterol-rich and oxidized cholesterol-rich diets). After 14 weeks to induce MI, rats in 3 groups were received ISO (100 mg/kg) for 2 consecutive days subcutaneously. Lipid profiles, electrocardiogram patterns and hemodynamic parameters of all groups were investigated.Serum levels of LDL, cholesterol and triglycerides were significantly higher in the fat-rich diet fed groups compared to control group (P<0.001). The ISO-treated rats showed a marked reduction in the R-amplitude, R-R interval, LVSP, left ventricular contractility (LVdP/dtmax) and relaxation (LVdP/dtmin) as well as severe elevation in ST-segment and LVEDP value compared to the respective normal rats. High serum level of OxLDL resulted in significant exacerbation in the destructive effects of ISO on R-amplitude, R-R interval, LVSP, left ventricular contractility (LVdP/dtmax), relaxation (LVdP/dtmin), ST-segment and LVEDP values. Additionally, heart to body weight ratio as an index of myocardial edematous was also increased significantly. However, changes in these parameters in rats fed with cholesterol-rich diet were not significant.Generally, results indicated that the effects of high OxLDL level on electrocardiogram and hemodynamic parameter after MI was more reliable than effects of high LDL level.

Serum concentrations and hypoglycemic effect of gliclazide: crosspovidone solid dispersion on streptozotocin induced diabetic rats.

Gliclazide is practically insoluble in water and its GI absorption is limited by its dissolution rate. Our previously published works indicated that preparing gliclazide-crosspovidone solid dispersion in the drug/ carrier ratio of 1:1 using cogrinding technique is able to enhance drug dissolution rate. The coground of gliclazide-crosspovidone was administrated to the rats and the hypoglycemic effects of pure drug, a physical mixture and the coground were considered in 3 groups of rats weighing 200-250 g (n=6). The rats were made diabetic by single intravenous administration of streptozotocin (60 mg/kg). Each of the rats received a single dose of gliclazide (equivalent to 40 mg/kg) as pure drug, physical mixture and coground in an aqueous suspension. Glucose level was assessed via glucometer after collecting the blood samples from tail vein. Gliclazide concentration in plasma was assessed applying high pressure liquid chromatography. According to 1-way ANOVA, Student-Newman-Keuls test, the coground revealed enhanced hypoglycemic effects as well as higher serum gliclazide concentration relative to pure drug and its corresponding physical mixture in the all sampling times. The area under serum glucose concentration curve vs. time for the pure gliclazide, physical mixture and coground formulations were 3 090.5±79, 3 018.8±96 and 2 374.0±73 mg.h/dl, respectively. Correspondingly, their area under serum gliclazide concentration curve vs. time were 1 171.8±156.8, 1 379.5±96.2 and 4 827.7±637.5 µg.h/ml. It follows that; formulation of gliclazide-crosspovidone coground is able to improve oral absorption of the drug.

Synthesis of molecularly imprinted monolithic fibers for solid-phase microextraction of acetaldehyde from head-space of beverages stored in PET bottles.

Molecularly imprinted monolithic fibers were synthesized and evaluated for solid-phase microextraction (SPME) of acetaldehyde from head-space of beverages stored in poly(ethylene terephthalate) (PET) bottles. The fibers were prepared by co-polymerization of methacrylic acid and ethylene glycole diethacrylate as functional monomer and cross-linker, respectively. Acetaldehyde was used as template molecule during polymerization process, in order to leave specific cavities after leaching off from the co-polymer. Narrow bore (i.d. 100 µm) glass capillaries as molds were filled with polymerization mixture containing cross linker, functional monomer, template molecule, initiator and porogen solvent. This mixture is then polymerized to form a continuous porous monolith that conforms to the shape of the mold. The resulting monolithic fibers were employed by a micro-syringe to extract acetaldehyde from head-space of sample solutions. Gas chromatography/mass spectrometry (GC/MS) was used to analyze adsorbed acetaldehyde to the fibers. Parameters influencing adsorption of acetaldehyde from head-space of standard solutions to the fiber and subsequently desorption in the GC chamber were optimized. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.01 and 0.03 µg mL(-1), respectively. The intra-day and inter-day precisions of the peak areas for five replicates were 8 and 15%, respectively.

Molecular characterization of a mucin-type antigen associated with human pancreatic cancer. The DU-PAN-2 antigen.

This work describes the molecular characterization of a human pancreatic cancer-associated antigen defined by a murine monoclonal antibody (DU-PAN-2). DU-PAN-2 antigen was isolated from a pancreatic adenocarcinoma cell line (HPAF) or patient's ascitic fluid, and the antigenic activity was monitored by competitive inhibition radioimmunoassay. Affinity chromatography and CsCl/guanidine HCl density gradient centrifugation were employed to remove other populations of mucin-type glycoproteins and noncovalently associated proteins, respectively. Three electrophoretically distinct components were detected by 1% agarose gel electrophoresis and were resolved by chromatography on Sepharose CL-4B. The major fraction (FII) was subjected to carbohydrate and amino acid analyses. The sum of threonine, serine, proline, glycine, and alanine comprised more than 50% of the amino acid residues. The saccharide units, O-glycosidically linked to the peptide via GalNAc, contained fucose, galactose, GlcNAc, GalNAc, and sialic acid. The total carbohydrate content of FI and FII was 80.8% and 77.4% by weight, respectively. The molecular weight of FII antigen showed two species of molecules of 1.45 X 10(6) and 4.59 X 10(6) by analytical sedimentation equilibrium. DU-PAN-2 antigen was susceptible to neuraminidase, pepsin, Pronase, and papain digestion. These results suggest that both protein components and sialic acid residues may play important roles in the binding of DU-PAN-2 antibody.

Aberrant expression of monoclonal antibody-defined colonic mucosal antigens in inflammatory bowel disease.

Human proximal colon from patients with inflammatory bowel disease and from controls was studied by two techniques to detect tumor-associated antigen expression. A panel of four murine monoclonal antibodies that recognize tumor-associated antigens was used to test purified colonic mucins for epitope expression by radioimmunoassay and to test formalin-fixed, deparaffinized sections of colon by the immunoperoxidase technique. The panel included monoclonal antibodies 19-9, B72.3, DU-PAN-2, and CSLEX1. Colonic mucins were purified from uninvolved surgical specimens by gel filtration with Sepharose 4B and cesium chloride-guanidine hydrochloride density gradient ultracentrifugation. Purified mucins from uninvolved colonic mucosal specimens from 4 of 7 patients with ulcerative colitis expressed one or more of these epitopes by radioimmunoassay, whereas mucins from 6 disease controls did not. Reactivity patterns were heterogeneous. Immunoperoxidase testing demonstrated staining with two or more antibodies in 14 of 18 involved inflammatory bowel disease segments, whereas control sections rarely stained with these antibodies, with the exception of 19-9. Sections of uninvolved mucosa from 4 of 9 patients with ulcerative colitis stained with two or more antibodies. Staining patterns were heterogeneous. The results demonstrate that colonic expression of tumor-associated epitopes occurs frequently in involved segments from both patients with ulcerative colitis and with Crohn's disease, whereas only patients with ulcerative colitis frequently expressed these epitopes in uninvolved segments.