Immunomodulatory effects of IFN-beta1a treatment alone or associated with pentoxifylline in patients with relapsing-remitting multiple sclerosis (RRMS).

Interferon-beta1a (IFN-beta1a) and pentoxifylline (PTX) are reported to be active in relapsing-remitting multiple sclerosis (RRMS), but the mechanisms are not completely understood. In two groups of RRMS patients, we studied the phenotype of peripheral lymphocytes and the level of several cytokines both in sera and in supernatants of activated peripheral blood mononuclear cells (PBMC) before and after 8 months of therapy with IFN-beta1a alone or associated with PTX. Our data indicate that patients with RRMS, treated with IFN-beta1a, exhibited a significant increase in CD4(+)CD25(++) T suppressor cells, accompanied by a significant decrease in cytotoxic lymphocytes (CD8(+)CD28(-) and natural killer [NK] cells) and IFN-gamma production, which could both contribute to an explanation of the previously described beneficial effects of IFN-beta treatment in MS. The addition of PTX to IFN-beta1a treatment did not modify the immunomodulatory effects obtained with IFN-beta1a alone. Future studies are needed to demonstrate which immunologic parameters correlate with the clinical benefit of IFN-beta1a treatment.

Comparison of fasting plasma leptin concentrations in healthy subjects with high and low plasma insulin.

This study was initiated to evaluate the role of hyperinsulinemia in the regulation of fasting plasma leptin. We measured plasma leptin and insulin concentrations in 404 healthy nondiabetic subjects. For analytical purposes, the population was divided into quartiles on the basis of the lowest (quartile 1) and highest (quartile 4) plasma insulin response to oral glucose, and fasting plasma leptin values in these 2 dichotomous groups were compared. The total plasma integrated insulin response was 4-fold greater in quartile 4, associated with significantly higher (P < .001) fasting plasma leptin (12.60+/-0.85 v8.53+/-0.56 ng/mL). Fasting plasma leptin concentrations remained significantly higher in the hyperinsulinemic quartile when comparisons were made after subdividing the population on the basis of gender, body mass index (BMI), or waist to hip ratio (WHR). These results demonstrate that fasting plasma leptin concentrations are significantly higher in hyperinsulinemic individuals, and this difference is independent of either overall or central obesity.

Attenuated expression of 70-kDa heat shock protein in WI-38 human fibroblasts during aging in vitro.

We examined the effects of cellular aging on the expression of the heat shock-inducible HSP70 gene in WI-38 diploid human fibroblasts serially passaged in vitro. The senescence of the cells was established by evaluating population doubling level, cell density at confluency, and cell morphology along with the detection of senescence-associated beta-galactosidase activity (histochemically detectable at pH 6), a reliable marker of aging in low-density cultures. A marked decrease in the synthesis and accumulation of the inducible HSP70 protein was observed in serum-fed late passage cells exposed to a severe heat shock (30 min at 45 degrees C) in comparison to early passage cells. However, the degree of HSF-DNA binding, monitored by gel retardation assay was similar in both early and late passage cells. Similarly, Northern blotting analysis indicated that comparable amounts of inducible HSP70 mRNA were present in the total RNA fraction, in the total polyadenylated RNA fraction, or in the nuclear polyadenylated RNA fraction extracted from both early and late passage cells. In contrast, much less inducible HSP70 mRNA was detected in the total cytoplasmic RNA fraction or in the polyadenylated cytoplasmic RNA fraction of late passage cells. Thus age-related differences in heat-induced HSP70 synthesis and accumulation observed in serum-fed WI-38 cells appeared to result from an impairment in the posttranscriptional processing of the HSP70 mRNA at a level following the polyadenylation step and preceding translocation from the nucleus to the cytoplasm. When HF were serum deprived for 20 h before heat shock, the induction of HSP70 mRNA was less than 30% reduced in early passage cells in comparison to serum-fed cells; however, the level of HSP70 mRNA was markedly (over 80%) decreased in serum-deprived late passage cells. This result indicated that the presence of serum has a strong influence on heat shock-induced HSP70 gene expression in human fibroblasts aging in vitro.

Clinical observations with a new specific assay for bone alkaline phosphatase: a cross-sectional study in osteoporotic and pagetic subjects and a longitudinal evaluation of the response to ovariectomy, estrogens, and bisphosphonates.

The purpose of this study was to examine the serum levels of bone alkaline phosphatase (BALP) measured with a new assay in normal and in osteoporotic women, and to evaluate prospectively its responsiveness to changes of bone metabolism. The following groups of subjects were studied: (1) 95 healthy women (44-75 years) (22 pre- and 73 postmenopausal) and 35 osteoporotic women [vertebral bone mineral density (BMD) more than 2.5 SD below the normal adult mean]; (2) 10 women (44-50 years) ovariectomized (OVX) for benign uterine diseases, examined before and 12 months after surgery; (3) 16 OVX women (36-54 years), examined before and after 12 months of transdermal estrogen replacement therapy (50 microg/day); (4) 12 previously untreated pagetic patients (4 women and 8 men, 50-80 years), examined before and 3 months after the I.V. administration of clodronate (600 mg) or alendronate (5 mg) for 2 consecutive days. The median BALP value was 11.6 U/liter (25-75th percentiles: 10.5-12. 7; range 7.7-19.3) in healthy premenopausal (PreMP) women and significantly higher (median: 16.8 U/liter; 25-75th percentile: 13. 8-21.8; P < 0.01) in postmenopausal (PostMP) women. There was a clear age-related increase in normal subjects (r = 0.43; P < 0.001). In the osteoporotic group, BALP levels, as well as other biochemical parameters of bone turnover, were not significantly different from those of normal women when adjusted for age. In OVX women, BALP levels showed a marked increase 12 months after surgery (median: 113%; 25-75th percentile: 87-139%), significantly higher than the increase of total ALP (median: 43%; 25-75th percentile: 25-66%; P < 0.001), and similar to the increases of serum osteocalcin and urinary hydroxyproline. Transdermal estrogen treatment prevented the BALP increase, even if no reduction was observed; total ALP showed a similar behavior. The basal levels of BALP were significantly elevated in pagetic patients (median: 91 U/liter; range 18-610 U/liter) and correlated to the scintigraphic extent of the disease (r = 0.76; P < 0.01). Three months after the I.V. administration of bisphosphonates, the decrease of BALP was more marked than that of total ALP (median: -54% versus -41%; P < 0.05). In conclusion, these results suggest that BALP measurement with this immunoassay may be clinically useful, and more sensitive than total ALP, in the assessment of bone turnover during changes of the estrogen status as well as in monitoring the effects of treatments that modify the metabolic activity of the skeleton.

Acute effects of bisphosphonates on new and traditional markers of bone resorption.

Bisphosphonates are known to be potent inhibitors of osteoclast activity and their only clinically relevant effect in the short-term is the selective inhibition of bone resorption. The purpose of this study was to compare the response to the intravenous administration of two bisphosphonates, clodronate and alendronate, of several biochemical markers of bone resorption, including tartrate-resistant acid phosphatase (TRAP) and cross-linked carboxyterminal telopeptide of collagen I (ICTP) in serum and hydroxyproline (OHP), free pyridinium cross-links (Pyr), and cross-linked N-telopeptides of collagen I (NTx) in urine. The study was carried out on 11 osteoporotic and 12 Pagetic subjects of both sexes, treated with clodronate (600 mg/day for 2 days) or alendronate (5 mg/day for 2 days), and monitored for 28 days after bisphosphonate administration. All the urinary markers of bone resorption showed a prompt decline after bisphosphonates, with maximum reductions after 7-14 days: Pyr decreased by 43% +/- 9% and 42% +/- 22% (mean +/- SD), respectively in osteoporotic and pagetic subjects, OHP by 51% +/- 14% and 51% +/- 20%, and NTx by 55% +/- 15% and 65% +/- 26%. In the osteoporotic group, the urinary markers began to increase again at 30 days, though still remaining well below the basal level, whereas in the pagetic group, the excretion of all markers remained depressed until the end of the observation period. The reduction of NTx was significantly greater than that of Pyr and OHP in pagetic patients (P < 0.05) and tended to be greater than that of Pyr in osteoporotic patients (p = 0.07).(ABSTRACT TRUNCATED AT 250 WORDS)

Acute coronary occlusion secondary to radiofrequency catheter ablation of a left lateral accessory pathway.

A case of asymptomatic acute coronary occlusion secondary to radiofrequency catheter ablation of a left lateral accessory pathway is reported. Due to post-procedural ST modifications of the surface ECG, a coronary angiography was performed which disclosed total occlusion of the first marginal branch of the left circumflex coronary artery. Acute myocardial infarction was confirmed by moderate cardiac enzyme release, abnormal myocardial perfusion scan and mild lateral hypokinesia at echocardiography. This rare but potentially harmful complication of interventional electrophysiology should be kept in mind and coronary angiography performed immediately when coronary occlusion related to radiofrequency application is suspected.

Effect of an alkaline shift on induction of the heat shock response in human fibroblasts.

The simultaneous exposure of WI-38 human fibroblasts (HF) to a heat shock (45 degrees C, 30 min) and an alkaline shift (> or = pH 8.0) in the incubation medium increased and extended the expression of heat shock proteins (hsps). Hsp70 was the most prominent inducible hsp synthesized during the recovery phase after the double shock, and the increase in synthesis depended on the degree of alkalinization during the heat shock. The accumulation of inducible hsp70, which was shown by Western blotting to occur in the late part of the recovery period, was more pronounced in the cells exposed to alkaline medium during the heat shock. Northern blotting did not reveal any increase in hsp70 mRNA, although time course studies following the double shock indicated a more prolonged presence of mRNA. Hsp70 gene activation was evaluated by a gel retardation assay using a 32P-labelled DNA oligonucleotide containing the heat shock consensus element (HSE) and a heat shock-induced specific binding protein (heat shock transcription factor, HSTF) from the cell extract. Heat shock activated HSTF-DNA binding and induced hsp70 mRNA expression as well as the synthesis and accumulation of hsp70. Alkaline shift, which by itself did not induce hsps expression, activated HSTF DNA-binding. However, in combination with heat shock, alkaline shift enhanced and prolonged HSTF-HSE complex association and hsp expression at both mRNA and protein levels. Since the alkaline shift-induced activation of hsp gene does not allow full transcription, these results provide further support for the multistep nature of the heat shock transcriptional response.

Different HSP70 expression and cell survival during adaptive responses of 3T3 and transformed 3T3 cells to osmotic stress.

Responses both to hyperosmotic stress and to heat shock were compared in 3T3 cells, spontaneously transformed cells (ST3T3) and simian virus 40-transformed cells (SV3T3). Cell adaptation to these stresses was measured in terms of surviving cell viability and plating efficiency, while their induced synthesis of stress proteins was monitored in terms of the presence of mRNA for HSP70, the pattern of polypeptides synthesised and the accumulation of HSP70 detectable by monoclonal antibodies. All three types of cells responded similarly to heat shock in their expression of HSP70 and showed no clear differences in ability to recover. In contrast, both ST3T3 and SV3T3 cells adapted more poorly and much more slowly to hyperosmotic stress (0.5 osM incubation) than did normal 3T3 cells. This different pattern of adaptation to hyperosmotic stress was parallelled by the cells' different expression of a stress protein that could not be distinguished from the heat-induced HSP70 by any of the methods listed above. In view of these findings it seems possible that hyperosmotic treatment might be useful in selectively affecting the survival of tumour cells.

Acute hemodynamic effects of intravenous amiodarone in patients with coronary artery disease.

The acute hemodynamic effects of intravenous amiodarone (Cordarone injectable; Labaz) were studied during cardiac catheterization in 16 male patients with coronary artery disease (age range, 38-64 years; mean, 53 years). Amiodarone was administered as a bolus at a dosage of 5 mg/kg bodyweight over a 1-min period. Measurements were made 5, 10, and 15 min thereafter. The drug had little effect on heart rate, aortic pressure, cardiac index, and vascular resistances. There were small and nonsignificant increases in left ventricular end-diastolic pressure and end-diastolic volume. The ejection fraction decreased slightly and not significantly. In addition to some increases in pulmonary wedge, pulmonary artery, and right atrial pressures, the significant findings were a 15% decrease in maximal dP/dt and a 12% decrease in left ventricular work. These changes point to a slight negative inotropic effect of amiodarone.

[Evaluation of a delayed-action beta blocker by 24-hour ambulatory recording of the arterial pressure]. / L'évaluation de l'effet d'un beta-bloqueur retard par l'enregistrement ambulatoire de 24 heures de la pression artérielle.

Twelve students aged 21 to 39 years (mean age 26) with borderline hypertension underwent three 24 hours ambulatory recordings of blood pressure using an Avionics Pressurometer II device: the first recording under basic conditions, the second after taking placebo and the third after being treated for 3 days with a betablocker of long acting effect (oxprenolol R 160 mg) in a randomized study. The blood pressure mean profile showed a double peak during daily activity with a progressive lowering of the blood pressure during sleep. The effect on the blood pressure of long acting oxprenolol begins four hours later, remains for about ten hours and may be still present next day. The shape of the circadian blood pressure profile is not modified. No significant difference between the pressure profiles on the basic conditions and placebo is noticed.

[Evaluation of a method for ambulatory and automatic measurement of nyctohemeral arterial pressure]. / Evaluation d'une méthode de mesure ambulatoire et automatique du profil nycthéméral de la pression artérielle.

This study was designed to evaluate a method of automatic non-invasive ambulatory blood pressure monitoring. Using a Del Mar Avionics Pressurometer II, we tested 12 normal subjects at rest and during exercise in our laboratory, and 12 patients with borderline hypertension during 24 hours of normal activity. At rest correlation coefficient of sequential measurements between pressurometer and a standard mercury sphygmomanometer were 0.96 for individual systolic measurements, 0.65 for individual diastolic measurements, and 0.99 for sequentially coupled diastolic or systolic measurements. The same correlation coefficients were found during a low level exercise test on cycloergometer (50 watts); at a higher level (75 watts), the correlation coefficient for systolic measurements is still high (0.93), whereas for diastolic measurements the correlation coefficient is low (0.45) in our experimental conditions. Fifty-eight non-invasive ambulatory recordings were obtained during normal daily activities. About 10% of the measurements were eliminated because of artefacts. We conclude that the nycthemeral fluctuations of the blood pressure can be reasonably estimated by the Del Mar Avionics Pressurometer II, but that such results can be obtained only if special attention is given to the positioning of the pressurometer on a motivated patient.