Rodolfo C. Puche

Inhibitory effect of fluoride on the secretion of insulin

SummaryThe oral administration of sodium fluoride (NaF) (40 μmol/100 body weight [bw]) to fasting rats produced an immediate fall in insulin levels and the consequent increase in glycemia. These phenomena were observed with plasma fluoride concentrations 5–15 μM. Glycemia and insulin returned to normal levels within 4–5 hours, together with the washing out of fluoride from plasma and soft tissues. The insulin secretion of isolated Langerhans islets, perifused with solutions containing 5, 10, or 20 μM fluoride, was found to be significantly inhibited as a function of fluoride levels, both with basal and stimulatory concentrations of glucose. One hour after the intake of 60 mg of NaF, fasting human volunteers showed increased fluoride (5–15 μM) together with a significant fall of plasma insulin levels.

Bone mass increase and glucose tolerance in rats chronically treated with sodium fluoride

Female rats were treated with fluoride for 100 days (between 21 and 121 days of age) replacing the water supply with a 5 mM NaF solution. Bone mass was assessed by destructive physical and chemical measurements on the whole skeleton, that gave an overall view not reported previously. Bone mass (dry, fat-free weight of the skeleton/100 g of body weight) increased 7% (P less than 0.001) with respect to control animals. This phenomenon was equally evident in the head, the axial and the appendicular skeleton. Fluoride treatment did not affect the ratio ashes/organic matrix. Treated animals showed a subtle disturbance of glucose tolerance as shown by glucose tolerance tests. The disturbance was manifest as high plasma and soft tissue levels of fluoride during the period of bone mass increase. Glucose tolerance was normalized when the maximum bone mass was achieved and plasma and soft tissue fluoride returned to control levels.

Calcium metabolism of rats with varying degrees of insulinopenia

This paper reports an investigation designed to determine the influence of varying degrees of insulinopenia upon the calcium metabolism of actively growing, alloxan-treated rats fed diets with three levels of calcium. A significant reduction in the skeletal mass (in absolute terms) was observed one month after alloxan administration in rats fed diets with normal or high calcium contents. The impact of insulin deficiency was greater on bone collagen than on the mineral mass, as shown by the increased calcium/hydroxyproline ratio. Alloxan-treated rats showed rather increased levels of PTH which was at variance with respect to control animals and unrelated to the calcium content of the diet. In spite of the high PTH levels, diabetic rats showed significantly diminished rates of bone Ca accretion and resorption. In addition, the animals fed the diet with the normal Ca content, showed significantly reduced areas of osteocytes lacunae and hypocalcemia after 24 h of fasting. The overall information obtained indicates that, in the rat, insulin deficiency more pronouncedly affects organic matrix than mineral turnover. The diabetic state is characterized by an impaired response of bone tissue to physiological stimuli, which is attributed to defective cellular activity caused by insulin deficit. Diminished bone resorption is considered to be an adaptative response to preserve bone mass.

An articulated rotating brush for office endometrial evaluation of climacteric outpatients.

This paper reports the experience obtained in the last 8 years with an instrument (articulated rotative brush) that allows access to the entire endometrial surface, obtaining material for both histological and cytological study. The latter has the advantage of reducing the number of cases in which insufficient material frustrates endometrial assessment. In addition to the overall experience gained with this instrument, the proportion of agreement with anatomo-pathological diagnosis after hysterectomy in 61 patients was high (96%) and the unweighted kappa statistic (0.862) indicated very good accordance between these techniques. Comparison between the present method and hysteroscopic observations in 84 patients with perimenopausal bleeding revealed a general conformity of 0.65 and an unweighted kappa statistic of 0.368 (fair agreement). In a group of 72 cases with postmenopausal bleeding a general conformity value of 0.92 was obtained, with an unweighted kappa statistic of 0.84 (very good agreement). In 90% of the cases the procedure was well accepted. In 57.3% of the cases, no difficulties were encountered using the brushing technique; in 25.9% slight dilatation was deemed necessary, whereas in 16.8% technical difficulties could not be overcome and general anesthesia was indicated. A total of 769 outpatients were investigated, 75.3% of which had normal endometria, 12.7% benign hyperplasias, 2.6% precursor lesions, 1.7% suspect carcinoma, and 4.2% carcinomas. Insufficient material for diagnosis occurred in 3.5% of cases tested. The combined cytohistological and hysteroscopic assessment of the endometrium provides satisfactory results in the etiological diagnosis of peri and postmenopausal bleeding, and in the follow up of patients undergoing hormonal replacement therapy (HRT).

The effects of long term feeding ofSolanum glaucophyllum to growing rats on Ca, Mg, P and bone metabolism

The effects of the long term ingestion ofSolanum Glaucophyllum leaves (SG) by the rat was investigated in two series of experiments; the animals were fed a normal (for 8 weeks) or a low Ca diet (for 5 weeks).With both diets, the intestinal absorption of Ca was increased and the endogenous fecal Ca excretion was decreased by SG treatment.Added to a normal Ca diet, SG increased the urinary excretion of Ca, Mg and P and reduced the excretion of hydroxyproline and pyrophosphate. At the histological level, SG induced a higher rate of bone tissue synthesis on trabecular and endosteal surfaces. The bone content of hydroxyproline and citrate increased significantly. The total alkaline phosphatase activity of plasma decreased as a function of SG intake due to a decrease in the activity of the intestinal isoenzyme, which was not compensated by the increases in the bone isoenzyme activity.The Mg absorption was decreased by SG inducing lower Mg balances and lower plasma Mg levels.Added to a low Ca diet, SG increased the severity of the secondary hyperparathyroidism induced by the diet. The urinary excretion of hydroxyproline and the plasma alkaline phosphatase activity (both isoenzymes) were significantly increased. The Na and K content of bone decreased as a function of SG intake.45Ca kinetic experiments revealed that SG increased the rate of Ca resorption and the rate constants of the fast exchangeable Ca pool, in both diets.

Binding of monofluorophosphate to α2-macroglobulin and C3

After administering an oral dose of monofluorophosphate (MFP) to human beings or rats, a fraction of the drug appears in plasma that is bound to proteins, establishing a previously undetected compartment of nondiffusible fluoride. This article documents experiments performed in vitro, describing the binding of MFP to two plasma globulins: α2-macroglobulin and C3 (a β-globulin). MFP binds irreversibly to these proteins through a stable bond. MFP binds to purified α2-macroglobulin or to C3 with a molar ratio MFP: protein close to unity. MFP binding reduces significantly the biological activity of these proteins, which share in common a macrocyclic 4-residue ring thiolactone (Cys—Gly—Glu—Glu).The binding site of MFP is as yet unknown. Protein-bound MFP appeared in the plasma of volunteers during the 5–7 hours following intake. Peak concentration of protein-bound MFP and maximal reduction of α2-macroglobulin activity was observed 2 hours after intake. Clearance of protein-bound MFP coincided with the return of α2-macro-globulin to basal levels.

Effect of calcium on rat intestinal alkaline phosphatase activity and molecular aggregation

Two fractions of rat intestinal alkaline phosphatase (IAP) were detected by Western blot: 168 ± 6 and 475 ± 45 kDa. The low molecular weight fraction constitutes 43% of the isolated proteins exhibiting 82% of the enzymatic activity, and a heavier fraction constitutes 57% of the isolated proteins and has 18% of the enzymatic activity. Calcium produced an increase of the 475-kDa form to the detriment of the 168-kDa form. This work also describes the kinetic and structural changes of IAP as a function of calcium concentration. With [Ca2+] < 10 mmole/L, the Ca2+-IAP interaction fitted a binding model with 7.8 ± 4.4 moles of Ca2+ /mole of protein, affinity constant = 19.1 ± 8.4 L/mmole, and enzymatic activity increased as a linear function of [Ca2+] (r = 0.946 p < 0.01). On the other hand, with [Ca2+] >10 mmole/L the data did not fit this model and, the enzymatic activity decreased as a function of [Ca2+] (r = − 0.703 p < 0.05).

Severe hyperglycemia: a determinant factor for hypofiltration in alloxan diabetic rats.

Abstract.The relationships among glomerular filtration rate, renal plasma flow and extracellular fluid volume were investigated in control and severely hyperglycemic (442±33 mg/dl) untreated, alloxan diabetc rats. Most of diabetic animals showed significant lower values of inulin clearance (diabetics, 0.55±0.07 ml/min·100 g; controls, 0.97±0.04) and p-aminohippurate clearance (diabetics, 2.11±0.39 ml/min·100 g; controls, 3.93±0.25). Diabetic rats exhibited reduced efficiency in tubular Na+ reabsorption, increased urinary Na+ excretion (diabetics, 3.12±0.27 mEq/day; controls, 1.25±0.14) and diminished values of plasma renin activity (diabetics, 3.34±0.44 ng/ml·h; controls, 8.64±0.79). Significant negative correlations were found between glycemia and renal hemodynamic variables. Acute overload with glucose further decreased these variables in both groups: inulin clearance in diabetics vs. controls, 0.26±0.04 vs. 0.44±0.05 ml/min·100 g; p-aminohippuric acid clearance in diabetics vs. controls, 1.09±0.20 vs. 1.55±0.21 ml/min·100 g. We conclude that chronically hyperglycemic alloxan diabetic rats showed diminished glomerular filtration rates (inulin clearance), renal plasma flow (p-aminohippurate clearance) and extracellular fluid volume associated with urinary Na+ losses and alterations in the renin-angiotensin system. Decreased reninangiotensin system activity might reduce aldosterone secretion, which in turn could result in (succesively) urinary sodium loss, extracellular fluid volume contraction and reductions in glomerular filtration and renal plasma flow.

Effect of bicarbonate feeding on immobilization osteoporosis in the rat

A time course study of immobilization osteoporosis was carried out in an experimental model in which the hind half of the rat was paralyzed by spinal section between the 3rd and 4th lumbar vertebrae. The earliest changes in the femur were a reduction in water content and in bone volume (3 days after operation). A significant reduction in both hydroxyproline and phosphate content was observed 8 days after immobilization. Calcium, magnesium, sodium, citrate, and carbonate were found significantly decreased at 15 days. Bone potassium remained unchanged. The rate of calcium accretion and the size of the exchangeable pools, when expressed per unit of bone mass, were not significantly affected by treatment.Sham-operated rats fed a bicarbonate-enriched diet, showed increased rates of calcium accretion and bone calcium content. The latter was not accompanied by increases in phosphate and carbonate contents. Operated animals fed the same diet showed an increased rate of bone resorption attributed to an assumed higher rate of parathyroid hormone secretion induced by bicarbonate feeding.

Calcium metabolism of intact and thyroparathyroidectomized rats fed a bicarbonate enriched diet.

Rats fed a bicarbonate-enriched diet become alkalotic during their feeding period (ca. 10 h/day). Alkalosis produced a significant reduction in the plasmatic concentration of ionized calcium and increased parathyroid hormone secretion confirmed by concurrent hypophosphatemia and increased urinary excretion of cAMP. Three weeks of treatment produced, however, a significant reduction of bone Ca resorption. The increased endogenous fecal excretion of calcium counteracted the enhancement of true calcium absorption. No significant change in the skeletal calcium mass was observed. Thyroparathyroidectomized rats fed the alkaline diet reduced further their rate of bone Ca resorption and increased significantly their skeletal calcium mass.