Effects of one-year tofacitinib therapy on bone metabolism in rheumatoid arthritis.

Janus kinase (JAK) inhibitors are used to treat rheumatoid arthritis (RA). We assessed the effects of tofacitinib on bone density and bone markers in association with clinical and laboratory parameters in RA. Tofacitinib stabilized bone density and resulted in a positive balance of bone turnover. INTRODUCTION: Janus kinase (JAK) inhibitors emerged as new therapeutic options in rheumatoid arthritis (RA). We have little information on how it affects areal and volumetric bone mineral density (BMD) and bone turnover markers. The aim of this study was to assess the effects of 1-year tofacitinib therapy on bone metabolism in RA. METHODS: Thirty RA patients with active disease were treated with either 5 mg bid or 10 mg bid tofacitinib for 12 months. We determined DAS28, CRP, IgM rheumatoid factor (RF), and anti-cyclic citrullinated peptide (CCP) levels, as well as serum levels of sclerostin, osteocalcin (OC), P1NP, DKK-1, OPG, RANKL, and 25-hydroxy-vitamin D3. Areal and volumetric BMD were assessed by DXA and peripheral quantitative CT (QCT), respectively. RESULTS: Twenty-six patients (13 on each arm) completed the study. Tofacitinib was clinically effective by suppressing DAS28, CRP, and HAQ. This was accompanied by the attenuation of further bone loss. Tofacitinib therapy significantly increased OC, OPG, and vitamin D3, while decreased CTX levels (p < 0.05). Age and multiple bone markers (OC, CTX, P1NP, RANKL) inversely correlated with L2-4 and femoral neck BMD by DXA. CRP, DAS28, and RANKL inversely determined volumetric BMD by QCT. Age, CRP, anti-CCP, and DKK-1 influenced the effects of tofacitinib therapy on BMD changes. CONCLUSIONS: One-year tofacitinib treatment stabilized BMD in RA patients and resulted in a positive balance of bone turnover as indicated by bone biomarkers. Further studies are needed to evaluate the potential beneficial effects of JAK inhibitors on inflammatory bone loss.

Safety, clinical outcome, and fracture rate of femoropopliteal stenting using a 4F compatible delivery system.

OBJECTIVE: To determine the safety, clinical outcome, and fracture rate of femoropopliteal interventions using 4F stents. METHODS: Between January 2010 and December 2011, 112 symptomatic patients were treated by stent implantation. Ten patients were lost to follow up; therefore, 102 patients (62 men; mean age 66.4 ± 10.1 years) were retrospectively analyzed. The indication for femoropopliteal revascularization was severe claudication (Rutherford-Becker score = 3) in 63 (62%) patients and chronic critical limb ischemia (Rutherford-Becker score = 4-6) in 39 (38%). Follow up included palpation of peripheral pulses and measurement of ankle brachial index. In patients with suspected in-stent restenosis duplex ultrasonography was performed. In 2013, patients were asked to return for a fluoroscopic examination of the stents. RESULTS: 114 lesions (Trans-Atlantic InterSociety Consensus-C and D, n = 45) were treated with 119 stents (Astron Pulsar, n = 42; Pulsar-18, n = 77). Lesions were long (≥100 mm) in 49 cases and heavily calcified in 35. Stents were long (≥120 mm) in 46 cases. Ten stents were partially overlapped. The technical and clinical success rates were 100%. Two puncture related complications were noted, neither of which required surgical repair. Eleven patients died (myocardial infarction, n = 4; stroke, n = 2; cancer, n = 5) and nine patients underwent major amputation (above knee, n = 4). The primary patency rate was 83% at 6 months and 80% at 12 months. The primary assisted patency rate was 97% at 6 months and 94% at 12 months. The secondary patency rate was 86% at 6 months and 85% at 12 months. The prevalence of fractures was 26% (type III and IV, 10%) after an average follow up of 25 months. CONCLUSION: Femoropopliteal stenting using a 4F compatible delivery system can be accomplished with a low complication rate, acceptable fracture rate, and with similar 12 month patency and revascularization rates as their 6F counterparts.

Health risk factors and mortality in Pécs City, Hungary in the 1990s.

In the period of 1990-1994 an increase of all causes mortality for 35-74 years old males was observed both in Pécs and in all Hungary. From 1994 to 1997 the mortality decreased. Similar changes, but of smaller dimension, were observed in the female population. The increasing mortality of the early 1990s is attributed primarily to the extra psycho-social stress of this period. The data of the population survey at Pécs in 1995-96 were compared to the data of earlier surveys. The mean blood total cholesterol levels and the prevalence of smoking decreased from 1990 to 1996. The prevalence of hypertension and male obesity increased. Physical inactivity, unhealthy diet and lack of improvement of diet still represent significant health problems. High prevalence of increased gamma-glutamyl transferase indicate high prevalence of excess alcohol consumption. The risk factor profile of 18-25 year old males is very unfavourable. Smoking prevalence in females aged 26-35 years exceeds that of males of the same age group. Preventive efforts should be focused to young males and females.

Determination of residues of pyrethroid and organophosphorous ectoparasiticides in foods of animal origin.

Analytical methods were introduced for the determination of residues of ectoparasiticides containing pyrethroid and organophosphate active ingredients in foods. Milk and edible tissues of cows treated with three experimental ectoparasiticides (containing cypermethrin + diazinon, deltamethrin + diazinon and alphamethrin + diazinon, respectively) were assayed for the presence of active ingredient residues. Synthetic pyrethroid residues were not detected in any of the samples processed. Diazinon residues could only be detected in milk samples taken on the first day after treatment (0.005-0.025 mg/kg) and in liver and fat tissue samples taken on the day of slaughtering (0.12 and 0.01 mg/kg, respectively). Permethrin and propetamphos residues were determined in the skin, meat and liver of chickens kept on 'Blotic-B' treated litter and in eggs collected at different times after the treatment of layer houses. Permethrin residues could not be detected in any of the samples (< 0.01 mg/kg). Meat and fat tissues of chickens slaughtered on the day after treatment contained small amounts of propetamphos (0.003 and 0.02 mg/kg, respectively). In the case of chickens kept on the treated litter and slaughtered after one week, active ingredient was not detected in meat, but 0.006 mg/kg propetamphos was present in the fat. The residue content of other samples (liver, egg) was below the detection limit of the applied method at all sampling times. From the food toxicological point of view these pesticide combinations can be used safely if the recommended withdrawal period is observed between ectoparasiticide administration and slaughter.

Changes in the expression of Na+/K+-ATPase isoenzymes in the left ventricle of diabetic rat hearts: effect of insulin treatment.

Na+/K+-ATPase related strophanthidin sensitive 3-O-methylfluorescein-phosphatase activity, [3H]ouabain binding and expression of Na+/K+-ATPase subunit isoforms were measured in the left ventricle of the heart of normal and streptozotocin-diabetic rats with and without insulin treatment. Compared to control animals, the enzyme activity was 0.75 +/- 0.09 and 0.62 +/- 0.06 times lower in rats diabetic for 2 and for 4 weeks, respectively. This was associated with a proportional decrease of the [3H]ouabain binding sites. Immunoblots indicated a 0.76 +/- 0.08 and 0.61 +/- 0.08-fold decrease of alpha1, a 0.68 +/- 0.09 and 0.41 +/- 0.04-fold decrease of alpha2 subunit in 2- and 4-week diabetic rats, respectively relative to controls. Beta1 subunit decreased proportionally 0.71 +/- 0.07 and 0.38 +/- 0.06-fold, and beta2 decreased 0.75 +/- 0.08 and 0.31 +/- 0.06-fold, respectively. Northern blot analysis revealed a significant reduction in mRNA level of Na+/K+-ATPase subunit isoforms after 2 and 4 weeks of diabetes (for alpha1 66.2 +/- 8.2 and 55.9 +/- 7.8% of controls for alpha2 91.7 +/- 12.1 and 41.1 +/- 7.1% of controls and for beta subunit 93.4 +/- 11.1 and 49.8 +/- 6.8% of controls, respectively). Although, mRNA levels of isoform reverted to even higher levels than the control values after insulin treatment, insulin caused only a partial recovery of enzyme activity, [3H]ouabain binding capacity and protein expression. We have obtained evidence that in cardiac left ventricle there are more than one type of Na+/K+-ATPase alpha and beta subunit isoforms which are affected in diabetes and by insulin treatment. The time course of diabetes induced changes and the degree of involvement suggest that the Na+/K+-ATPase isoforms are altered individually.

Interference of the sulphonylurea antidiabeticum gliquidone with mitochondrial bioenergetics in the rat under in vitro conditions.

The hypoglycaemic sulphonylurea gliquidone and glibenclamide exerted a partial uncoupling effect on mitochondrial respiration of liver under in vitro conditions using various citrate cycle intermediates as substrates. Besides the uncoupling effect, gliquidone and glibenclamide caused a direct inhibition of ATP-as well as DNP-stimulated oxygen consumption. Both phenomena proved to be dose dependent. Respiratory control ratio decreased progressively with increasing concentrations of sulphonylureas mainly through the inhibition of ADP-stimulated respiration. Basal and DNP-stimulated ATP-ase activity of isolated mitochondria changed similarly to the respiratory parameters. Changes in membrane permeability of mitochondria and the inhibition of substrate uptake further support the assumption of structural and functional alteration of mitochondria by the hypoglycaemic compounds tested.

Interference of sulphonylurea antidiabetica with mitochondrial bioenergetics under in vivo conditions.

Sulphonylurea antidiabetica effectively inhibits the basal hepatic glucose production. Since it has been firmly established that lipophylic sulphonylurea drugs exerted an uncoupling effect on mitochondrial oxidative phosphorylation, a relationship between the reduction of hepatic gluconeogenesis and the insufficient energy supply due to sulphonylureas could be supposed. In this study we have investigated the effects of glibenclamide and gliquidone on mitochondrial bioenergetics in liver after peroral treatments of normal rats with different doses. The treatment of rats with 5 mg/kg glibenclamide or gliquidone daily for 14 days elicited only a marginal inhibition on mitochondrial oxidation capacity and remained without any effect on mitochondrial ATPase activity. Only the supermaximal dose 50 mg/kg for 14 day produced a significant damage in the mitochondrial functions. The basal respiration increased with 60-80 per cent, whereas the ADP- or DNP-stimulated oxygen consumption significantly decreased independently from the respiratory substrates investigated. Similar alterations were found in the mitochondrial ATPase activity after treatment with these drugs. No essential differences have been observed in the actions between glibenclamide and gliquidone. However, the lowest dose applied in this study is many times higher than the usual therapeutic dose. Consequently, glibenclamide and gliquidone do not interact with mitochondrial bioenergetic processes under therapeutic conditions. On the other hand, in different liver and kidney damages we have no sufficient knowledge whether these drugs can be accumulated in these organs and therefore their elevated concentration may interfere with the mitochondrial energy metabolism.

Effect of streptozotocin-induced diabetes on kidney Na+/K(+)-ATPase.

The maximal capacity of low affinity ouabain binding sites in kidney medulla was found to be increased by 20 +/- 3.8% after 2 weeks, and by 35 +/- 4.5% in 4 weeks diabetes. However, in kidney cortex no similar changes could be detected. Western blot analysis of Na+/K(+)-ATPase subunits in kidney medulla indicated a significant enhancement of both the alpha 1 and beta 1 subunit in two and four weeks diabetic rats (alpha 1: 35 +/- 3.1, 51 +/- 5.8% and beta 1: 31.3 +/- 5.2 and 43.2 +/- 6.8%, respectively). However, kidney cortex showed no significant change in any condition tested. In diabetes we could detect a significant change only in the medulla in case of the b subunit mRNA transcript, which showed 1.69 +/- 0.59 and 2.89 +/- 0.81 times increased in two and four weeks diabetic state, respectively. There was no change in the alpha 1 subunit mRNA abundance. Insulin treatment of diabetic animals did not result in a complete reversal of diabetes-induced changes in ouabain binding capacity or in the amount of Na+/K(+)-ATPase alpha 1 and beta 1 subunit protein and mRNA levels. Our data indicate a good correlation between changes in low affinity ouabain binding capacity and the level of alpha 1 isoform in diabetic rats, and suggest an important role of the b subunit in the regulation of enzyme quantity.

On the nature and molecular basis of prostacyclin induced late cardiac changes.

The late appearing and long-lasting cardiac effects induced by PgI2 or its stable analogue 7-oxo-PgI2 described by us first in 1983 include: 1.) antiischemic, 2.) antiarrhythmic, 3.) selective electrophysiological and 4.) cardiac cytoprotective changes. Evidence for 1.: Protection from myocardial ischemia due to coronary occlusion in dogs furthermore a significant diminution of ischemic loss of the myocardial ATP and CP content and of the myocardial lactate accumulation in excised rat heart exposed to global ischemia. 2: Protection from postocclusion and reperfusion arrhythmias in dogs. 3: A selective prolongation of the ventricular refractory period (VERP) as well as of the action potential duration (APD90) in the isolated rabbit papillary muscle furthermore prolongation of QT distance and VERP in the rabbit and the guinea pig heart "in situ". 4.: The cardiac cytoprotective effect, i.e. ischemic loss of intracellular K+ and ischemic gain of intracellular Na+ in isolated hearts of 7-oxo-PgI2 treated guinea pigs subjected to 25 min global ischemia was significantly moderated. These protective actions proved to be dose and time dependent - maximal effects appeared 48 hrs after administration of a single dose of 50 micrograms/kg i.m. 7-oxo-PgI2. These effects are certainly not due to 7-oxo-PgI2 itself but this latter seems to be indispensable for induction of a long acting principle, which can probably be extracted from hearts of pretreated animals and this substance of lipoid character exerts electrophysiological effects similar to those observed after 7-oxo-PgI2 pretreatment. Pretreatment may also reduce isoproterenol induced heart rate increase and prolong bleeding time in conscious rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of immune complexes from SLE patients on human monocyte locomotion and Fc receptor function.

The effect of immune complexes (IC) isolated from systemic lupus erythematosus (SLE) sera with polyethylene glycol and gel filtration on the chemotaxis and Fc receptor function of healthy monocytes was examined. Even at a low protein concentration (1 microgram/ml = 1 mg/l) ICs inhibit monocyte chemotaxis. ICs from patients with SLE nephritis are more inhibitory than ICs from patients without renal disease. The inhibitory effects of ICs on monocyte chemotaxis and Fc receptor activity are similar, suggesting a relationship between the chemotactic and Fc receptor function of monocytes. Analysis of the ICs by enzyme-linked immunoassay showed no correlation between the quantity of IgG, C3, and anti-DNA in the IC samples and their effects on monocyte function.

Stimulating effect of tuftsin and its analogues on the defective monocyte chemotaxis in systemic lupus erythematosus.

Monocytes and macrophages are engaged at various levels of cellular immune reactivity. In addition to their function in the defensive mechanism directed at infective agents, they also play a basic role in immune complex elimination and antigen handling. Previous experiments revealed that systemic lupus erythematosus (SLE), the main representative of the autoimmune diseases, is associated with impaired monocyte chemotaxis. The endogenous basic tetrapeptide tuftsin and 6 of its analogues were examined in vitro for their stimulating capacity on the chemotactic responsiveness of monocytes derived from patients with SLE. The monocyte migration assay was carried out by a modified Boyden technique and quantified by the leading front distance method and by counting the total distance covered by the monocyte locomotion. Tuftsin and 3 of its analogues significantly increased the defective chemotaxis in SLE. The tetrapeptides effective on chemotaxis also stimulated random migration and phagocytosis of the monocytes, albeit to a lesser extent. Structure-activity relationships, as well as the influence of the clinical stage of the disease were also examined. Experimental evidence leads to a favourable prediction for the immunotherapeutic value of these oligopeptides for the control of infections and the progression of the disease in patients with systemic lupus erythematosus.