Anti-phospholipase A2 Receptor Antibody in Differentiation and Prognostication of Membranous Nephropathy.

Anti-M-type phospholipase A2 receptor (anti-PLA2R) antibody is believed to be associated with primary membranous nephropathy (pMN) and absent in secondary MN (sMN). There are few data regarding utility of anti-PLA2R antibody as a prognosticator. Our study aimed to compare the incidence of positive serum anti-PLA2R antibody titer in pMN versus sMN and correlation with clinical outcome. From August 2015 to July 2019, patients with biopsy-proven MN were evaluated for serum anti-PLA2R antibody titers by the enzyme-linked immunosorbent assay. The subset of cases was repeated to monitor the clinical response in terms of 24 h proteinuria. A total of 169 patients, 65 pMN and 104 sMN were studied. Anti-PLA2R antibody was found in 41 (63.08%) pMN with mean titer, 232.62 RU/mL, and 40 (38.46%) sMN with mean titer 253.59 RU/mL. Out of positive antiPLA2R antibody titer in pMN cases, 15 were retested twice to 5 times with mean titers of 78.95, 36.27, 13.9, and 15.45 RU/mL, respectively. Out of positive anti-PLA2R antibody in sMN cases, 11 were retested twice to five times with mean titers of 104.42, 122.49, 12.33, and 17.2 RU/mL, respectively. All patients with decreasing anti-PLA2R antibody titer in both groups had clinical remission, with a decrease in mean 24 h proteinuria from 7.11 g to 3.36 g in pMN and 5.97 g to 3.41 g in sMN. Ten pMN and 11 sMN patients without remission showed persistent positive anti- PLA2R antibody titer. Anti-PLA2R antibody titer may be elevated in pMN/sMN. It can also be used as a noninvasive prognostic marker for MN.

Pharmacokinetic/pharmacodynamic evaluation of marbofloxacin as a single injection for Pasteurellaceae respiratory infections in cattle using population pharmacokinetics and Monte Carlo simulations.

Population pharmacokinetic of marbofloxacin was investigated with 52 plasma concentration-time profiles obtained after intramuscular administration of Forcyl® in cattle. Animal's status, pre-ruminant, ruminant, or dairy cow, was retained as a relevant covariate for clearance. Monte Carlo simulations were performed using a stratification by status, and 1000 virtual disposition curves were generated in each bovine subpopulation for the recommended dosage regimen of 10 mg/kg as a single injection. The probability of target attainment (PTA) of pharmacokinetic/pharmacodynamic (PK/PD) ratios associated with clinical efficacy and prevention of resistance was determined in each simulated subpopulation. The cumulative fraction of response (CFR) of animals achieving a PK/PD ratio predictive of positive clinical outcome was then calculated for the simulated dosage regimen, taking into account the minimum inhibitory concentration (MIC) distribution of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni. When considering a ratio of AUC0-24 hr /MIC (area under the curve/minimum inhibitory concentration) greater than 125 hr, CFRs ranging from 85% to 100% against the three Pasteurellaceae in each bovine subpopulation were achieved. The PTA of the PK/PD threshold reflecting the prevention of resistances was greater than 90% up to MPC (mutant prevention concentration) values of 1 µg/ml in pre-ruminants and ruminants and 0.5 µg/ml in dairy cows.

Urinary Screening for Early Detection of Kidney Diseases.

OBJECTIVE: Urinary screening is a simple inexpensive tool to evaluate kidney functions. The authors carried out urinary screening of school children for early detection of kidney diseases. METHODS: Children in the age group 5-15 y were screened for urinalysis. They were divided in 2 groups; group-1 included 5-10 y and group-2 included >10-15 y old children. RESULTS: Urine samples of 3340(78%) out of 4283 enrolled children were tested. Abnormal samples were found in 5.75%; with proteinuria in 4.59%, pyuria in 3.29% and hematuria in 4.31%. Males constituted 47.71% in group-1 and 54.64% in group-2. Low body mass index was found in 94.1% group-1 and 78.99% group-2 children. Mild proteinuria was found in 1.2% group-1 and 2.56% group-2 children. Severe proteinuria was more in group-2 (0.77% vs. 0.06%) with female preponderance. Glucosuria was found in 1 boy of group-2. Urobilinogen was more in group-2 (0.65% vs. 0.24%) with male preponderance. Nitrituria was found in 9 girls. Pyuria (2.02% vs. 1.27%) and hematuria were more in group-2 (3.04% vs. 1.87%) with female preponderance. Combined proteinuria and hematuria (0.42% vs. 0.24%) as well bacteruria and fungaluria were more in group-2 (4.11% vs. 1.39%). Six of 192 children with abnormal urinary findings were treated; 1 for urinary calculus and 5 for urinary tract infection. CONCLUSIONS: Abnormal urinary findings were more common in children >10 y of age. Thus urinary screening program of children can become useful for early detection of kidney diseases and contribute towards building up of a healthy nation.

A pharmacokinetic/pharmacodynamic model capturing the time course of torasemide-induced diuresis in the dog.

A pharmacokinetic/pharmacodynamic modelling approach was used to determine a dosage regimen which maximizes diuretic efficiency of torasemide in dogs. Kinetic profiles of plasma concentration, torasemide excretion rate in urine (TERU) and diuresis were investigated in 10 dogs after single oral administrations at 3 dose levels, 0.2, 0.8 and 1.6 mg/kg, and an intravenous injection of 0.2 mg/kg. Endogenous regulation was evidenced by a proteresis loop between TERU and diuresis. To describe the diuresis-time profile, TERU served as input into a turnover model with inhibition of loss of response, extended by a moderator acting on both loss and production of response. Estimated maximum inhibition of loss of response, Imax , was 0.984 showing that torasemide is an efficacious diuretic able to suppress almost total water reabsorption. A TERU50, value producing half of Imax , of 1.45 µg/kg/h was estimated from the model. Pharmacokinetic and pharmacodynamic parameters were used to simulate the torasemide dose-effect relationship after oral administration. Model predictions were in good agreement with diuresis measured in a validation study conducted in 10 dogs, which were administered oral doses of 0.15, 0.4, 0.75, 1.5 and 4.5 mg/kg for 5 days. Finally, oral dose associated with the highest daily diuretic efficiency was predicted to be 0.1 mg/kg.

Pharmacokinetics of marbofloxacin in pigs after intravenous and intramuscular administration of a single dose of 8 mg/kg: dose proportionality, influence of the age of the animals and urinary elimination.

The pharmacokinetics of marbofloxacin in pigs were evaluated as a function of dose and animal age following intravenous and intramuscular administration of a 16% solution (Forcyl(®) ). The absolute bioavailability of marbofloxacin as well as the dose proportionality was evaluated in 27-week-old fattening pigs. Blood PK and urinary excretion of marbofloxacin were evaluated after a single intramuscular dose of 8 mg/kg in 16-week-old male pigs. An additional group of 12-week-old weaned piglets was used for the evaluation of age-related kinetics. The plasma and urine concentration of marbofloxacin was determined using a HPLC method. Pharmacokinetic parameters were calculated using noncompartmental methods. After intravenous administration in 27-week-old fattening pigs, the total body clearance was 0.065 L/h·kg. After intramuscular administration to the same animals, the mean observed Cmax was 6.30 µg/mL, and the AUCINF was 115 µg·h/mL. The absolute bioavailability was 91.5%, and dose proportionality was shown within the dose range of 4-16 mg/kg. The renal clearance was about half of the value of the total clearance. The total systemic clearance values significantly decreased as a function of age, being 0.092 L/h·kg and 0.079 L/h·kg in pigs aged 12 and 16 weeks, respectively.

Tissue-specific alterations in lipoprotein lipase activity in the rat after chronic infusion of isoproterenol.

This study describes the consequences of sustained exposure to the nonspecific beta adrenergic agonist isoproterenol on the activity of lipoprotein lipase (LPL) in tissues that are sites for a large portion of the intravascular hydrolysis of triacylglycerols. Chronic exposure to isoproterenol was accomplished by means of osmotic pump infusion during four consecutive days, and assessment of the activity of LPL was performed at the end of the treatment. Four days of ISO infusion resulted in a large decrease in LPL activity of parametrial white adipose tissue, which declined to half of that found in the untreated group. In contrast, brown adipose tissue LPL activity was greatly increased (approximately threefold) by ISO treatment. The beta agonist raised LPL activity in red vastus lateralis muscle by a factor of four over control values, whereas heart enzyme activity was not significantly altered by the drug. These results demonstrate that catecholamines are part of the factors which determine the tissue specificity of LPL regulation. The tissue-specific changes in enzyme activity in response to isoproterenol, which are in very good agreement with the known metabolic actions of catecholamines in adipose and skeletal muscle tissues, attest to the key role of the enzyme in the partitioning of lipid substrates in situations of increased sympathetic activity.

Serum free fatty acids are not involved in acute exercise-induced reduction of LPL in rat tissues.

The present studies were designed to verify whether preventing the rise in serum levels of nonesterified fatty acids (NEFA) by adrenergic blockade would interfere with the decrease in tissue lipoprotein lipase (LPL) activity caused by acute exercise in rats. Ninety minutes before being killed, male rats were injected intraperitoneally with either saline, the beta-adrenergic blocker propranolol (25 mg/kg body weight), or the alpha 2-adrenergic blocker yohimbine (3 mg/kg body weight). Half of each group was killed at rest and the other half immediately after a 1-h run on a treadmill. LPL was determined in white adipose tissue (WAT), heart, and red vastus lateralis muscle (VLM). Exercise enhanced serum levels of NEFA 50% over resting values in saline-injected rats. The latter increase was totally abolished in animals having received propranolol or yohimbine. The activity of LPL in WAT, heart, and red VLM was approximately 35% lower in exercised rats than in resting animals. Serum triacylglycerols were also reduced by the run. Neither propranolol nor yohimbine interfered with any of these reductions. Exercise did not change serum glucose levels in saline-injected rats but decreased it in those injected with propranolol or yohimbine. Serum insulin was unchanged by exercise and by the antagonists. These findings suggest that the beta- and alpha 2-adrenergic pathways, as well as the exercise-induced rise in serum levels of NEFA, are not responsible for the early reducing effect of a 1-h run on tissue LPL activity in untrained rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Beta-adrenergic blockade and lipoprotein lipase activity in rat tissues after acute exercise.

The present experiments were aimed at evaluating the acute effects of exercise on lipoprotein lipase (LPL) activity in untrained rats. The activity of LPL was measured in postheparin plasma (PHP) before and at various times after a 1-h run on a treadmill (22 m/min, O degrees grade). LPL in PHP was 50% below pre-exercise levels immediately and 3 h after the run but was increased 65% over resting levels 24 h postexercise. To further characterize the very early fall in LPL activity in response to exercise and to assess the possible involvement therein of the beta-adrenergic pathway, LPL in heart, vastus lateralis muscle (VLM), and white (WAT) and brown (BAT) adipose tissues was determined at rest and immediately after exercise in rats that were treated or not with nadolol (25 mg.kg-1.day-1 for 30 days). Immediately after 1 h of exercise, there was a reduction in total enzyme activity in WAT (40% below resting levels), BAT (-58%), VLM (-53%), and heart (-30%). Exercise reduced serum triacylglycerol levels (-64%) and doubled those of nonesterified fatty acids. beta-Adrenergic blockade did not affect any of these variables. Both exercise and nadolol lowered serum cholesterol levels by approximately 20%, but the effects were not additive. These results show that the global intravascular pool of LPL undergoes divergent, time-dependent alterations in response to a single bout of moderate exercise. The acute downregulation of postheparin plasma LPL immediately after exercise reflected a fall in the total enzyme pool of all tissues studied.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute effects of exercise on triacylglycerol metabolism in fasted, untrained rats treated with nadolol.

The current study investigated the early response of some determinants of triacylglycerol (TG) metabolism to acute exercise in rats that were treated or not with the nonselective beta-adrenergic blocker, nadolol (25 mg.kg b.wt.-1.day-1 for 30 days). Measurements of hepatic TG secretion rate (HTGSR), postheparin plasma hepatic TG lipase (HTGL) activity, and that of lipoprotein lipase (LPL) in postheparin plasma, heart, vastus lateralis muscle (VLM), white (WAT) and brown (BAT) adipose tissues were carried out in untrained rats at rest or immediately after a 1 h run on treadmill (22 m.min-1, 0 degrees grade). All animals were in the fasted state. Exercise reduced serum TG levels (46% below resting levels) and doubled those of nonesterified fatty acids. HTGSR was enhanced (+28%) by exercise while HTGL activity was not modified. The decrease in postheparin plasma LPL activity (-24%) caused by exercise was consistent with a reduction in enzyme activity in WAT (-32%), BAT (-45%) and heart (-25%). One h of treadmill running did not influence LPL activity in VLM. beta-Adrenergic blockade did not affect any of the variables of lipid metabolism, except for a slight decrease in specific activity of LPL in the heart (-14%). This study demonstrates that absolute variations in HTGSR and in LPL activity cannot account for the acute fall in serum TG levels caused by moderate exercise, in fasted, untrained rats. In addition, these results show that the beta-adrenergic pathway is either not involved in, or not necessary for, the latter effects of exercise on TG metabolism.

Restoration of lipoprotein lipase activity in insulin-deficient rats by insulin infusion is tissue-specific.

The activity of lipoprotein lipase was measured in white and brown adipose tissues, red vastus lateralis muscle, and heart of rats that have been insulin deficient (streptozotocin, 75 mg.kg-1) for 2 weeks, and that have then received implants of insulin-delivering minipumps (17 U.kg-1.day-1) for 1 or 4 days. Normal glycemia was restored in insulin-deficient animals after 4 days of insulin treatment. Hypertriglyceridemia, but not hypercholesterolemia, was reversed after 4 days of insulin infusion. After 2 weeks of insulin deficiency, fasting lipoprotein lipase activity was lowered in all tissues studied. In white adipose tissue, lipoprotein lipase decreased to 50% of control values. After a single day of insulin infusion, even if tissue weight has not yet been greatly affected, total activity was completely restored to control levels. Enzyme activity in brown adipose tissue was also depressed in deficient animals, and insulin infusion was followed by a slow recovery of activity, to a level intermediate between those of control and insulin-deficient groups. Insulin status had milder effects on lipoprotein lipase activity in vastus lateralis muscle than in the adipose tissues. Deficient rats displayed 60% less activity than controls, and 4 days of hormone infusion only partially restored enzyme activity. There was a large loss of lipoprotein lipase in the heart following 2 weeks of insulin depletion, which was not counteracted by hormone infusion. Thus the speed and extent of recovery of lipoprotein lipase activity following hormone replacement in insulin-deficient animals varied widely among tissues. These findings suggest that insulin is part of the factors that determine the tissue specificity of lipoprotein lipase regulation.

Isozymes of Superoxide Dismutase in Mitochondria and Peroxisomes Isolated from Petals of Carnation (Dianthus caryophyllus) during Senescence.

The balance between reactions involving free radicals and processes which ameliorate their effect plays an important role in the regulation of plant senescence. In this study a method was developed to isolate peroxisomes and mitochondria from carnation (Dianthus caryophyllus L. cv Ember) petals. Based on electron microscopy and marker enzyme levels, the proportion of peroxisomes to mitochondria increases during senescence. The superoxide dismutase (SOD) content of these fractions was examined. Mitochondria and peroxisomes were shown to contain two electrophoretically distinct SODs, a manganese-, and an ironcontaining SOD. The Mn- and Fe-SOD were found to have relative molecular weights of 75,000 and 48,000 and isoelectric points of 4.85 and 5.00, respectively. The presence of a Fe-SOD in mitochondria and peroxisomes is unique because this enzyme is usually located in chloroplasts. The activity of these two isoenzymes decreased during senescence in mitochondria but remained high in peroxisomes from senescent tissue. It is suggested that peroxisomes play a particular role in the process of senescence.

Mutagenicity of basic fractions derived from lamb and beef cooked by common household methods.

Mutagen production was examined in lamb and beef in relation to certain common household cooking methods. Mutagenicity was assessed, after extraction of the basic fraction of cooked meat samples, using Salmonella typhimurium strain TA1538 with added rat-liver S-9 homogenate. Little or no mutagenicity was found in barbecued lamb chops, in microwave-cooked lamb chops, sirloin steak, leg of lamb, or rolled beef loaf, in roasted leg of lamb or rolled beef loaf, in stewed blade steak or in boiled chuck steak. However, the basic fraction from well-done, edible fried or grilled meat contained mutagenic activity equivalent to approximately 30,000 TA1538 revertants/100 g cooked meat. It was found tht the mutagenic activity of grilled lamb chops, sirloin and rump steaks was directly related to the average surface temperatures attained during cooking. Use of butter as a frying medium was particularly associated with higher mutagenicity in meat samples. Fried meats (rump and fillet steaks) generally yielded higher mutagenic activity than did grilled meats (rump steak, lamb chops) at comparable temperatures of the cooking medium. Using similar cooking procedures, lamb did not differ markedly from beef in mutagenic activity.

[Comparison of amoxicillin and amoxiclav in the therapy of respiratory infections]. / Uporedenje amoksicilina i amoksiklava u terapiji respiracionih infekcija (RI).

Randomly hospitalized patients with respiratory tract infections admitted to three pulmonary departments of the Golnik Institute for Pulmonary Diseases and Tuberculosis were enrolled in an open, comparative clinical study of Amoksiklav and Amoxicillin. A group of 26 patients with a mean age of 64.5 years presenting with pneumonia (13), exacerbation of chronic bronchitis (12) and bronchiectasis (1) were given Amoskilav, while another 20 patients with a mean age of 61.4 years presenting with pneumonia (9), exacerbation of chronic bronchitis (5), bronchiectasis (5) and sinusitis (1) received Amoxicillin. The efficacy of treatment was assessed by bacteriological findings of respiratory tract specimens, sputum and blood leucocytosis, macroscopic purulence of sputum and the presence of fever. The bacteriological findings are shown in detail. Leucocytosis and macroscopic purulence of sputum significantly improved on Amoksiklav therapy (p less than 0.05) while with Amoxicillin there was no significant improvement. With respect to the presence of fever, there was no significant difference between Amoksiklav and Amoxicillin. The overall clinical and bacteriological response was very good and good in 88.5% of patients treated with Amoksiklav compared to 75% of those receiving Amoxicillin. Additionally, 1000 pathogenic strains were tested for their response to Amoksiklav and Amoxicillin. Amoksiklav proved superior against strains of Branhamella catarrhalis, E. coli, coagulase-negative staphylococci and K. pneumoniae (p less than 0.01).

Effect of hepatic vagotomy on postexercise substrate levels in food-restricted rats.

The metabolic effects of a selective hepatic vagotomy (HV) were investigated at rest and immediately after a 50-min exercise period (26 m/min, 0% grade) in rats subjected to an overnight 50% food restriction. This dietary restriction reduced liver glycogen content to 50% of normal resting concentrations (2.2-2.8 g/100 g). No significant differences between HV and sham-operated rats were found in resting and exercising beta-hydroxybutyrate, glucose, glycerol, and insulin concentrations. Postexercise liver glycogen concentrations were reduced to approximately 1.0 g/100 g in both HV and sham-operated groups. This decrease was associated with significantly (P less than 0.01) lower postexercise glycogen levels in the soleus muscle of HV rats (2.6 times) along with higher plasma free fatty acid concentrations (P less than 0.01). These data provide evidence that HV combined with a progressive decrease in liver glycogen content may influence substrate regulation during exercise. They also support the concept of the existence of hepatic glucoreceptors responsive to a decrease in liver glycogen content.

Identification of different classes of superoxide dismutase in carnation petals.

Isoelectric focusing and nondenaturing two-dimensional electrophoresis, followed by an enzymatic characterization involving use of specific inhibitors were applied to identify the different forms of the superoxide dismutase (SOD) extracted from petals of cut carnation. Electrophoresis was carried out either with the crude extract or with fractions obtained after ion exchange chromatography of the extract. Three forms of the SOD were identified (Cu,Zn-SOD, Fe-SOD and Mn-SOD). They have close pI's (4.7 to 4.9) and most of them are composed of several isoforms.

Effect of selective hepatic vagotomy on plasma FFA levels in resting and exercising rats.

Metabolic effects of a selective hepatic vagotomy (HV) were investigated in nonfasted (N) and 24-h fasted (F) rats, at rest and immediately after a 50-min exercise period (26 m/min, 0% grade). In nonfasted rats, no significant differences between HV and sham-operated (SHM) groups were found in blood substrates [free fatty acids (FFA) or glucose], insulin, and muscle glycogen levels, either at rest or after exercise. In F rats, liver glycogen was almost completely depleted at rest. This depletion was associated with a significantly (P less than 0.05) lower plasma FFA concentration at rest in HV compared with SHM rats (mean +/- SE, 0.57 +/- 0.04 vs. 0.83 +/- 0.1 mmol/l). No differences in FFA levels were observed between the same two groups after the exercise period. Exercise, however, resulted in a 2.5 times greater glycogen breakdown in the soleus muscle of HV compared with SHM rats. Hepatic vagotomy in the F condition was also associated with lower resting and exercising insulin concentrations. It is concluded that HV, when followed by a 24-h fast, may influence metabolic substrate regulation at rest and to a certain extent during exercise. These data support the concept of the existence of hepatic glucoreceptors responsive to a decrease in liver glycogen content.