Effects of ipriflavone and its metabolites on a clonal osteoblastic cell line.

Protective effects of ipriflavone, an isoflavone derivative, in osteoporosis are believed to be caused by the inhibitory action on bone resorption. A direct effect of ipriflavone on bone formation is as yet unknown. Ipriflavone and four of its metabolites (I, II, III, and V) were examined for their effects on parathyroid hormone response, collagen synthesis, alkaline phosphatase activity, and cell proliferation in a clonal cell population of rat osteoblastic cells. Pretreatment of osteoblasts with high concentrations of ipriflavone for 48 h significantly inhibited the cAMP response to parathyroid hormone, producing a shift in the dose-response curve; at lower concentrations metabolites II and III potentiated the cAMP accumulation induced by low doses of parathyroid hormone. The 48 h treatment with metabolite V at the 1 nM dose significantly stimulated collagen synthesis in osteoblastic cells. Ipriflavone and metabolite I showed a biphasic stimulatory action on the alkaline phosphatase activity of osteoblasts, with a maximal effect at the 0.1 and 1 nM doses, respectively. A similar biphasic response was observed with ipriflavone and metabolite I on osteoblastic cell growth, with a maximal effect at the 0.1 nM concentration. These results suggest a direct role of ipriflavone in modulating the synthetic and growth properties of osteoblast-like cells.

Effect of ipriflavone--a synthetic derivative of natural isoflavones--on bone mass loss in the early years after menopause.

OBJECTIVE: We studied whether oral administration of ipriflavone, a synthetic derivative of naturally occurring isoflavones, could prevent bone loss occurring shortly after menopause. DESIGN: Fifty-six women with low vertebral bone density and with postmenopausal age less than five years were randomly allocated to receive either ipriflavone, 200 mg three times daily, or placebo. All subjects also received 1,000 mg elemental calcium daily. RESULTS: Vertebral bone density declined after two years in women taking only calcium (4.9 +/- 1.1%, SEM, p = 0.001), but it did not change in those receiving ipriflavone (-0.4 +/- 1.1%, n.s.). A significant (p = 0.010) between-treatment difference was evidenced at both year 1 and year 2. At the end of the study, urine hydroxyproline/creatinine excretion was higher in the control group than in the ipriflavone group, as compared to no difference at baseline. Five patients taking ipriflavone and five taking placebo experienced gastrointestinal discomfort or other adverse reactions, but only one and four subjects, respectively, had to discontinue the study. CONCLUSIONS: Ipriflavone prevents the rapid bone loss following early menopause. This effect is associated with a reduction of bone turnover rate.

A cross-over controlled study on beclobrate versus bezafibrate in the treatment of type IIb hyperlipoproteinaemia.

The effects of the new fibric acid derivative beclobrate (100 mg) on some plasma lipidic and apoproteic parameters in 20 patients with type IIb hyperlipoproteinaemia were matched in a randomized cross-over study to a sustained release formulation of bezafibrate (400 mg). Inclusion criteria were: total cholesterol (TC) plasma levels greater than 260 mg/dl and triglyceride (TG) levels greater than 200 mg/dl, after a 2-month period of isocaloric diet. The drugs were administered once a day for 8 weeks, and then crossed-over after 8 weeks of wash-out for another 8 week period of treatment. Both drugs were significantly effective in reducing TC and TG, and in increasing HDL-C plasma levels, but only beclobrate significantly decreased Apo B and LDL-C whereas bezafibrate seemed to be more active in increasing HDL-C levels. The electrophoretic pattern of LP in patients treated with beclobrate revealed a decrease of VLDL, a normalization of the LDL fraction and an increase of HDL. The tolerability was generally good.

Treatment of acute pain of ureteral and biliary colic with naproxen sodium administered by the parenteral route.

Non-steroidal, anti-inflammatory agents (NSAIDs), wellknown inhibitors of prostaglandins, have been used in the treatment of biliary and ureteral pain since the end of the 1970s. The efficacy and tolerance of a new injectable formulation of naproxen sodium in ureteral and biliary pain was investigated in 77 out-patients, observed in an emergency ward, and affected by acute lithiasic symptomatology. Forty-four patients received one 275 mg vial of naproxen sodium intramuscularly, while 33 patients were given one vial at the same dosage intravenously. In 56% of the cases complete relief of pain was achieved within 30 minutes of injection, while in 86% pain was completely relieved or greatly decreased within one hour. Side-effects (nausea, vomiting) occurred in three patients, but were linked to a simultaneous aggravation of the ureteral colic.

Pharmacokinetic study on piroxicam at the steady-state in elderly subjects and younger adults after administration of piroxicam beta-cyclodextrin.

The age effect on the kinetics of piroxicam at the steady-state after oral administration of piroxicam beta-cyclodextrin was evaluated. The mean plasma concentration of piroxicam at the steady-state was significantly higher in elderly subjects (9.30 +/- 0.69 micrograms/ml mean +/- s.e.) than in younger adults (6.24 +/- 0.58 micrograms/ml mean +/- s.e.). Even though the distribution volume tended to be lower in elderly subjects (106.37 ml/kg), it did not show substantial differences in the two groups whereas the correlation between clearance and age was significant (p less than 0.05). It is therefore confirmed that piroxicam beta-cyclodextrin has the same pharmacokinetic behaviour at the steady-state in elderly subjects, as the active principle in a non-complexed form.

Sobrevivência de operários do cupim-de-montículo Cornitermes cumulans (Kollar, 1832) (Isoptera: Termitidae) alimentados com diferentes dietas artificiais / Survival of workers of the termite Cornitermes cumulans (Kollar, 1832) (Isoptera: Termitidae) fed with different diets

Avaliou-se a sobrevivência de operários do cupim Cornitermes cumulans (Kollar, 1832), isolados da colônia e mantidos em diferentes dietas artificiais em laboratório (21 ± 1° C; 90±10% UR e 24h de escotofase), em Lavras, Minas Gerais, Brasil. Nos bioensaios ofereceram-se ou não as dietas a 50 indivíduos por placa de Petri plástica com tampas perfuradas. Foram utilizadas seis repetições por tratamento, avaliando-se diariamente a sobrevivência dos cupins até a morte de todos os indivíduos. Os resultados obtidos foram submetidos à análise de sobrevivência de Weibull e as equações submetidas ao teste de identidade de modelos não-lineares. Houve diferença na sobrevivência estimada dos cupins tratados com as diferentes dietas testadas. Concluiu-se que é possível manter operários de Cornitermes cumulans vivos e isolados da colônia com o emprego de dieta contendo papel filtro, bagaço de cana-de-açúcar e ágar como fonte de alimento para esses cupins.

We evaluated the survival of the termite Cornitermes cumulans (Kollar, 1832), isolated from the colony and kept on different artificial diets in the laboratory (21 ± 1° C, 90 ± 10% UR and 24h of scotophase), in Lavras, Minas Gerais, Brazil. Fifty termite workers were isolated in each plastic Petri dish, where they were offered diets or not. We used 6 replicates per treatment, evaluating the daily survival of termites until the death of all the individuals. The results were submitted to Weibull survival analysis and the equations were subjected to the test of identity of non-linear models. There was a difference in estimated survival of termites treated with different diets. It was concluded that it is possible to keep Cornitermes cumulans workers alive and isolated from the colony with the use of a diet containing filter paper, bagasse cane sugar and agar as a food source.

Efficacy of ipriflavone in established osteoporosis and long-term safety.

Ipriflavone (i.p.), an isoflavone derivative, is currently used in several countries for prevention and treatment of osteoporosis. Recently, 149 elderly, osteoporotic women (65-79 years) with prevalent vertebral fractures were enrolled in two Italian, multicenter, double-blind, 2-year studies. Women were randomly allocated to receive either oral i.p. (200 mg T.I.D. at meals) or matching placebo, plus 1 g oral calcium daily. One hundred eleven subjects completed the 2-year treatment period. A significant increase in forearm bone mineral density (BMD), measured by dual photon absorptiometry (DPA), was obtained after i.p. treatment. Women receiving the placebo showed only a limited bone loss during the treatment period, probably due to calcium supplement; however, a significant between-treatment difference was obtained in both studies. Urinary hydroxyproline was significantly decreased in i.p.-treated patients, suggesting a reduction in bone turnover rate. A reduction of incident vertebral fractures was observed in i.p.-treated women compared with control subjects. A significant improvement of bone pain and mobility has also been pointed out in one of the studies. To date, 2769 patients have been treated with i.p., for a total of 3132 patient/years, in 60 clinical studies performed in Italy, Japan, and Hungary and reviewed for long-term safety assessment. The incidence of adverse reactions in ipriflavone-treated patients (14.5%) was similar to that observed in subjects receiving the placebo (16.1%). Side effects were mainly gastrointestinal. Few patients presented reversible modifications of laboratory parameters. The data from the above studies show that long-term treatment with i.p. may be considered safe, and may increase bone density and possibly prevent fractures in elderly patients with established osteoporosis.

Prevention of early postmenopausal bone loss using low doses of conjugated estrogens and the non-hormonal, bone-active drug ipriflavone.

Hormone replacement therapy is the optimal therapeutic choice for postmenopausal syndrome. While low doses of estrogens (0.3 mg/day of conjugated estrogens) can counteract neurovegetative menopausal symptoms, higher doses (0.625 mg/day of conjugated estrogens) are required to prevent bone loss in postmenopausal women. Experimental and clinical studies have shown that ipriflavone, a non-hormonal isoflavone derivative, is effective in the prevention and treatment of postmenopausal osteoporosis. The aim of the present investigation was to evaluate the efficacy and tolerability of ipriflavone and very low doses of equine conjugated estrogens on bone loss in early postmenopausal women. Eighty-three healthy postmenopausal women (50.3 +/- 0.7 years) were enrolled for this 1-year multicenter study. All subjects were randomly allocated to receive: double placebo (n = 24; group A), placebo plus conjugated equine estrogens 0.30 mg/day (n = 31; group B) or conjugated equine estrogens 0.30 mg/day plus oral ipriflavone 200 mg tris in die at meals (n = 28; group C), according to a double-masked design. Among women who completed the treatment period (valid completers), those of group A showed a progressive decrease in forearm bone density (FBD; measured by dual photon absorptiometry) that reached 1.7% after 12 months. The women in group B maintained their FBD in the first 6 months of treatment but, at the end of the study, showed a bone loss of 1.4% compared with basal values. By contrast, women in group C showed a significant increase in FBD after 1 year of treatment (+5.6%; p < 0.01). Both valid completers and intention to treat analyses revealed a significant difference (p < 0.05) between group A and group C over the study period. None of the treatments produced significant changes of biochemical markers of bone turnover, while hot flushes and other climacteric symptoms were significantly reduced after the sixth month of treatment in women receiving estrogens. Adverse events were generally mild, and did not differ among the groups. The results of this study suggest that low doses of estrogens combined with ipriflavone could represent a new therapeutic approach to the treatment of the postmenopausal syndrome.

Placebo-controlled comparison of piroxicam-beta-cyclodextrin, piroxicam, and indomethacin on gastric potential difference and mucosal injury in humans.

The acute gastroduodenal mucosa injury and gastric potential difference (GPD) drops provoked by 14-day administration of 20 mg/day of a new piroxicam formulation (piroxicam-beta-cyclodextrin), 20 mg/day standard piroxicam and 100 mg/day indomethacin were evaluated and compared in a randomized, double-blind, placebo-controlled study carried out on 64 volunteers. Endoscopic examinations, performed after 14-day treatment, demonstrated that piroxicam-beta-cyclodextrin was less gastrolesive (mean endoscopic score +/- SE = 0.56 +/- 0.2) than either piroxicam (2.06 +/- 0.5) or indomethacin (2.25 +/- 0.5) (p < 0.01). The drop in GPD after a single dose of the assigned drug was considerably greater for piroxicam and indomethacin than for piroxicam-beta-cyclodextrin (p < 0.01), which registered similar values to placebo. Since GPD is an expression of the anatomo-functional integrity of the gastric barrier, the results indicate that piroxicam-beta-cyclodextrin exerts less direct acute damage on the gastric mucosa. Therefore, when administered short-term, piroxicam-beta-cyclodextrin appears to be less gastrolesive than either indomethacin or the standard piroxicam formulation.

Does measurement of amniotic fluid index detect changes in amniotic fluid volume after second-trimester amniocentesis?

The objective of this study was to establish whether variations of amniotic fluid volume induced by second-trimester amniocentesis could be detected by serial measurements of amniotic fluid index. A total of 130 singleton pregnancies undergoing second-trimester amniocentesis for genetic indications were considered. Amniotic fluid index was measured at three different time intervals: 30-60 min before amniocentesis, immediately after the procedure, and 60 min after the procedure. Serial measurements were obtained either by a single operator (n = 55) or by the three independent operators (n = 75). Significantly lower amniotic fluid index values were demonstrated immediately after amniocentesis when compared with the pre-amniocentesis and subsequent measurements in the study design with both the single and multiple operators. No statistically significant changes were found between the first amniotic fluid index measurements and those obtained 1 h after amniocentesis. These results suggest that second-trimester amniocentesis induces a temporary decrease of amniotic fluid volume detectable by serial amniotic fluid index measurements, no longer evident 1 h after the procedure.

Fetal cardiac and extracardiac flows preceding intrauterine death.

Extracardiac and cardiac flow velocity waveforms were recorded in a severely growth-retarded fetus 1 day and a few hours before fetal death. At the first scan, the typical Doppler patterns of a growth-retarded fetus were found, but the brain-sparing effect was lost at the last examination and a huge tricuspid insufficiency was demonstrated.

Ipriflavone inhibits osteoclast differentiation in parathyroid transplanted parietal bone of rats.

Ipriflavone, a synthetic isoflavone-derived flavonoid, was shown to have inhibitory effect on bone resorption. In order to study its mechanism of action directly on bone, 46 female Wistar rats were divided into six groups and medicated orally for 25 days as follows: groups 1 and 2 were given 1% carboxymethylcellulose solution (vehicle), groups 3, 4, 5, and 6 were administered ipriflavone at doses of 0.178, 0.356, 0.712, and 1.424 mmol/kg/day (suspended in vehicle), respectively. On the 22nd day, parathyroid glands, taken from donor rats, were transplanted in contact with the outer surface of the periosteum of both the right and the left parietal bones of rats from groups 2, 3, 4, 5, and 6. The group 1 rats underwent sham operation. Bone histomorphometry, performed on the ectocranial periosteum of parietal bones, showed that absolute erosion boundary, absolute eroded area, absolute erosion depth, number of tartrate-resistant acid phosphatase (TRAP)-positive polynucleated osteoclasts, and number of TRAP-positive mononucleated cells decreased in ipriflavone-treated rats compared with group 2 rats. The reduction was roughly proportional to the increase of drug dosage and reached statistical significance in rats of groups 5 and 6. The same parameters were extremely low in group 1 rats. Mineral apposition rate did not differ in any of the groups. Significant increase of serum calcium and significant decrease of serum phosphate were found in group 2 rats compared with group 1 rats, whereas no differences from controls were detected in ipriflavone-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Ipriflavone does not alter bone apatite crystal structure in adult male rats.

We have previously found that a short-term treatment with high doses of ipriflavone increased bone density and improved the biomechanical properties of adult male rat bones, without altering their mineral composition. To determine whether this effect can be associated with alterations of bone crystal structure, we have performed X-ray diffraction analysis of bones obtained from rats treated with ipriflavone at doses that were effective in inducing favorable changes on bone density and biomechanics. Eighteen-week-old male Sprague Dawley rats were treated by oral route with either ipriflavone (200 or 400 mg/kg/day), or its vehicle for 12 weeks. The treatment was well tolerated and body weight increased to the same extent in all animals. As a measure of bone crystallinity, we examined the (310) and (002) reflections of the X-ray diffraction patterns, corresponding to the directions perpendicular and parallel to the c-axis of the crystals, respectively. No major differences were observed between ipriflavone-treated and control animals for the broadening parameter beta(1/2) for (310) and (002) peaks, as well as for lattice parameters. Therefore, a 12-week treatment with ipriflavone at high doses does not induce significant modifications of bone "crystallinity." Thus, the positive effect of ipriflavone on bone mineral density appears to be associated with an increased apatite crystal formation rather than an increase of crystal size. These results provide further evidence for the safety and usefulness of ipriflavone in the treatment of osteoporotic syndromes.

[Clinical and electrophysiologic changes produced by (--) eburnamonine in acute and post-acute stages of head injuries]. / Modificazioni cliniche ed elettrofisiologiche indotte dalla (--) eburnamonina nei traumatizzati cranici in fase acuta e post-acuta.

Two groups of patients suffering from cranial trauma have been submitted to a double-blind acute (1-2 per phlebo ampules/die for 7-10 days) and chronic (1 i.m. ampule for 30 days) at random treatment, respectively with (--) Eburnamonine and Papaverine. The evaluation of the clinical symptoms (state of consciousness, neurologic and post-traumatic symptoms) and of some electrophysiological responses (REG, visual and somato-sensorial evoked potentials) were investigated in basal conditions, during the acute stage, at the end of the chronic treatment and 30 days after the drugs withdrawal. The results showed (--) Eburnamonine to induce a superior improvement than Papaverine of some clinical symptoms (motor disorders, retrograde amnesia, vertigo) and of cortical bioelectrical activity, as rheoencephalographic data and reduction in early latencies of evoked response to somato-sensorial stimulation (SEP) revealed. Local and systemic tolerability was good with both drugs; a slight hypotensive action, more marked with Papaverine, was noted at the end of the treatment.

Effects of acute and chronic treatment with (-)eburnamonine on the tissue supply of oxygen.

Recent research has demonstrated the major role of oxygen in the maintenance of normal cerebral function in the elderly and considerable importance has been attributed to the action of intra-erythrocytic 2-3 diphosphoglycerate (2-3 DPG) in favouring the availability of oxygen to the tissues. This property is reportedly helped or potentiated by the pharmacological action of some eumetabolic vasoregulator drugs. According to numerous clinical trials, (-)eburnamonine possesses this property. A two-part study was carried out in elderly volunteers: a double-blind randomized trial on 20 patients treated acutely (10 with an infusion of 100 mg (-)eburnamonine vs 10 controls) and a within-patients crossover trial on 10 patients treated chronically by the oral route. The results showed a significant increase in the 2-3 DPG values and a beneficial modification of P50 after (-)eburnamonine, probably dependent both on the dose and mode of administration. The effect of the drug was much more marked but more fleeting after acute administration and less intense but more lasting after chronic administration. The latter mode of treatment is of interest in view of the therapeutic effects on several symptoms of senile chronic cerebral insufficiency.

Comparison of faecal blood loss, upper gastrointestinal mucosal integrity and symptoms after piroxicam beta-cyclodextrin, piroxicam and placebo administration.

In order to evaluate the gastric tolerance of the new piroxicam formulation CHF 1194 (piroxicam complexed with beta-cyclodextrin), a double-blind randomized trial was carried out in 21 young healthy volunteers comparing CHF 1194 with piroxicam and placebo. Faecal blood loss measurement by the Cr-51 labelled red blood cell technique, upper gastrointestinal endoscopic evaluation, titration of gastric pH and gastric biopsies before, during and after treatment were used to assess drug tolerability. Four out of 7 volunteers in the piroxicam-treated group withdrew because of severe gastrointestinal symptoms and oesophageal and/or gastroduodenal lesions, while all subjects treated with CHF 1194 or placebo completed the treatment. There was a significant difference between the endoscopic scores of the piroxicam and placebo groups, whereas no differences were found between CHF 1194 and placebo, nor between piroxicam and CHF 1194. Daily mean gastrointestinal blood loss was greater in the piroxicam group than in either the CHF 1194 or placebo groups, but the difference was not significant, due to the small number of piroxicam-treated subjects who completed the study. When administered for a short period to healthy young subjects, CHF 1194 caused less gastric damage and was better tolerated than piroxicam.

Effects of 1-year treatment with ipriflavone on bone in postmenopausal women with low bone mass.

Ipriflavone (IP) (7-isopropoxyisoflavone), a synthetic isoflavone derivative, is active in both inhibiting bone resorption and enhancing osteoblast function. This property suggested its clinical use in the treatment of involutional osteoporosis, and in the prevention of postmenopausal bone mass loss. Forty postmenopausal women with low bone mineral content were enrolled and randomly treated for 12 months with IP 600 mg/day or placebo (PL), according to a double-blind, parallel group design. All patients wee also given an oral calcium supplementation (1 g/day). Bone mineral density (BMD) was measured at the spine (L2-L4) by dual-energy X-ray absorptiometry and at the distal radius by single-photon absorptiometry. Bone metabolism markers (serum calcium, phosphate, osteocalcin, and alkaline phosphatase, and urinary calcium, phosphate, and hydroxyproline) were assessed at the same times. After 12 months, a reduction of BMD was evidenced in the PL-treated group, at both the spine (-2.2%, P < 0.01 vs baseline) and the forearm (-1.2%). In the IP-treated group, an increase of BMD was obtained (+1.2%, P < 0.01 vs placebo, at the spine; +3%, not significant, at the forearm). Bone markers were in the normal range for postmenopausal women; no statistically significant modifications were observed during the treatment period. Three patients were withdrawn from the treatment in the IP-treated group, and two in the PL-treated group for gastrointestinal disturbances. In the other women, the tolerance of the drug was good and the compliance with the oral treatment was excellent.

Lack of any estrogenic effect of ipriflavone in postmenopausal women.

Estrogen replacement therapy (ERT) has been demonstrated to prevent osteoporosis in postmenopausal women (PMW). However, several contraindications exist for ERT and many PMW cannot be treated. It has also been shown that too low doses of ERT are able to exert therapeutical effects on some climacteric symptoms but not on bone and compounds exerting synergic actions with ERT on bone without effects on other organs could be useful. The isoflavone derivative, ipriflavone, seems to have this effect but data are lacking on its endocrine effect in humans; thus, this study was undertaken to clarify in PMW whether ipriflavone exerts estrogenic activity. Evaluation of LH and FSH secretion during a 24-h period was performed in a group of 15 PMW after a single oral dose of 600 or 1,000 mg of ipriflavone or placebo, and after 7, 14 and 21 days of oral treatment with ipriflavone 600 mg and 1,000 mg/daily, administered in three divided doses. LH secretion was also evaluated during naloxone infusion before and after 21 days of ipriflavone, placebo or conjugated estrogen treatment (0.625 mg/day; CE). LH response to NAL treatment was absent during ipriflavone and placebo such as it was observed before treatments. By contrast, a significant increase of LH plasma levels was measured during naloxone infusion in CE-treated women. This result demonstrates that ipriflavone is unable to exert the same effects that estrogens do in PMW. In addition, no changes like in placebo group were seen on vaginal cytology in this group of subjects after 21 days, whereas a significant increase of superficial vaginal cells was observed after 21 days of CE treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytological and ultrastructural investigation on osteoblastic and preosteoclastic cells grown in vitro in the presence of ipriflavone: preliminary results.

The effects of ipriflavone on bone cells were studied in vitro on pre-osteoclastic (FLG 29.1) and osteoblast-like (Saos-2) cells grown for 48 h either separately or in co-cultures, with or without the addition of PTH. Histological, ultrastructural and histochemical (TRAP-activity demonstration) methods were used. The main results show that ipriflavone reduces replication of FLG 29.1 cells and inhibits TRAP production by these cells both in controls and in co-cultures treated with PTH. Moreover, it has a moderate stimulatory effect on proliferation of osteoblast-like cells and reduces the PTH-induced degenerative changes of Saos-2 cells. These results suggest that the inhibitory effect of ipriflavone on FLG 29.1 cells might be indirect and might be mediated by the osteoblast-like cells.

Effect of ipriflavone on bone mass in elderly osteoporotic women.

A study in elderly osteoporotic women was performed to assess the effect of one year treatment with ipriflavone (IP) on bone mass and bone biomarkers. Twenty-eight women aged over 65, with diagnosis of osteoporosis and X-ray evidence of at least one vertebral fracture, were treated with IP tablets (600 mg/day) or placebo (PL), according to a randomized, double-blind, parallel-group design. One g/day calcium supplementation was given to all patients. After 12 months a significant increase (+6%, P < 0.05) of bone mineral density (BMD) at the distal radius (DPA) was obtained in the IP-group. Serum osteocalcin (BGP) and urinary HO-proline/creatinine (HOP/Cr) values were reduced in the same group. BMD values did not change (-0.3%) in the placebo group. One woman of the PL-group was withdrawn from treatment because of worsening of pain, due to new vertebral crushes. Side effects (mainly gastrointestinal) arose in 8 IP- and in 5 PL-treated women. The compliance to the oral administration was good.