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Biomedical subjects

E Bonucci

Publications and source records attributed to E Bonucci.

At least 19 recordsLinked to original sources

Matrix vesicles and focal proteoglycan aggregates are the nucleation sites revealed by the lanthanum incubation method: a correlated study on the hypertrophic zone of the rat epiphyseal cartilage.

Correlated studies were performed with light and electron microscopy, and backscattered electron image in conjunction with X-ray microanalysis, of lanthanum-incubated epiphyseal cartilage of the young rat. The hall-mark of this procedure is the appearance of LaP electron-dense deposits (not present in control sections) in precise sites of the hypertrophic zone. The ultrastructural study revealed a dual nature of these sites: "dense matrix vesicles" and "focal filament aggregates". The dense matrix vesicles are a specific type of matrix vesicle with the intrinsic capacity of precipitating LaP mineral, as soon as they originate from the hypertrophic chondrocytes. Furthermore, the matrix vesicles were found to be heterogeneous because lanthanum-devoid, "light matrix vesicles" were also present. The focal filament aggregates, which were not recognized in unstained sections and in controls, are apparently focal concentrations of proteoglycans with high lanthanum binding capacity, although the presence in them of other components (e.g., type X collagen, C-propeptide of type II collagen) cannot be excluded. The were in close connection with the light matrix vesicles in the upper hypertrophic zone, and were loaded with a variable quantity of LaP irregular electron-dense deposits in the lower hypertrophic zone. These irregular deposits are similar to, but distinct from, calcification nodules. The lanthanum incubation method indirectly detects the matrix Ca-binding components (which bind La ions), and the calcification initiation sites (which precipitate a LaP-mineral phase). A sequence is proposed of successive steps of LaP nucleation within the focal filament aggregates, which possibly mimics calcium phosphate deposition. Such a sequence seems to require the participation not only of dense matrix vesicles, but also of the filamentous components of the focal aggregates, possibly together with the activity of alkaline phosphatase.

Animals

Combined use of malachite green fixation and PLA2-gold complex technique to localize phospholipids in areas of early calcification of rat epiphyseal cartilage and bone.

Two cytochemical methods (malachite green fixation and PLA2-gold complex technique) were combined in order to detect the presence and ultrastructural distribution of phospholipids in epiphyseal cartilage and metaphyseal bone of 25-day-old rats. Chondrocytes and osteoblasts showed a more intense PLA2-gold complex labeling of rough endoplasmic reticulum than mitochondria and plasma membranes. Roundish osmiophilic, electron-dense, lightly labeled structures were visible at the periphery of the cells. In areas of early mineralization of cartilage, not all the matrix vesicles were labeled by PLA2-gold complex. Calcification nodules showed intense labeling in comparison with the surrounding, lightly labeled uncalcified matrix; gold particles were chiefly found at their periphery. Calcification nodules of metaphyseal bone were deeply electron-dense after glutaraldehyde-malachite green fixation; PLA2-gold labeling was mainly found in connection with crystals, whereas the uncalcified matrix was weakly labeled. Calcified bone matrix showed a heavy labeling with randomly scattered gold particles. When pre-embedding decalcification was carried out in the presence of malachite green, a good preservation of organic component was obtained. The PLA2-gold positivity of decalcified areas of cartilage and bone confirmed the presence of phospholipids in mineralized matrix.

Animals

Frequency of adynamic bone disease and aluminum storage in Italian uraemic patients--retrospective analysis of 1429 iliac crest biopsies.

BACKGROUND: Adynamic bone disease was initially attributed too aluminum intoxication in association with low circulating levels of parathyroid hormone. More recently adynamic bone disease has been described even in the absence of aluminum intoxication. PURPOSE OF THE STUDY: It was the purpose of this retrospective analysis of 1429 iliac crest biopsies sent to our laboratory from 1985 to 1994 by 41 Italian nephrology and dialysis centres to assess the frequency of adynamic bone disease and aluminum accumulation. METHODS: Adynamic bone disease was diagnosed by histological and histodynamic (tetracycline labelling) analysis, on the basis of predetermined criteria. Aluminum accumulation was assessed by aluminon histochemical staining. RESULTS: The frequency of adynamic bone disease was fairly constant at approximately 15% from 1985 to 1994. In contrast, aluminum accumulation, defined as positive aluminon histochemical staining, decreased during the same period from 36% to 4%. CONCLUSIONS: Our data clearly show a dissociation of the incidence of adynamic bone disease and aluminium accumulation in bone. At least today, given the low prevalence of aluminium intoxication, factors other than aluminium are the main cause of adynamic bone disease.

Aluminum

Renal bone disease in 76 patients with varying degrees of predialysis chronic renal failure: a cross-sectional study.

BACKGROUND: Renal osteodystrophy has been studied less extensively in predialysis than in dialysis patients. Different types or histological patterns in their natural evolution from moderate to advanced severity of renal insufficiency are only partially known, with special regard to adynamic bone disease and its relationship with osteomalacia. METHODS: We conducted a cross-sectional retrospective study on 76 unselected patients with chronic renal failure undergoing conservative treatment, with a wide range of severity of renal insufficiency. All the patients were subjected to bone biopsy for histological and histomorphometric evaluation. The patients, 44 males and 32 females ranging in age from 18 to 72 years and with serum creatinine 1.2-11.4 mg/dl, had not been exposed to aluminium-containing drugs and had never been treated with vitamin D or calcitriol. RESULTS: Ten patients had normal bone, nine were diagnosed with adynamic bone disease, 26 with mild mixed osteodystrophy, seven with predominant osteomalacia, 22 with advance mixed osteodystrophy, and two with predominant hyperparathyroidism. Patients with adynamic bone disease had less severe chronic renal failure than the other pathological subgroups, intact PTH above the upper limit of normal, normocalcaemia, and reduced serum osteocalcin in line with a significantly lower ObS/BS. Osteomalacia was found in a more advanced stage of chronic renal failure with relative hypocalcaemia and more severe metabolic acidosis. A creatinine clearance of 20 ml/min served as a clear demarcation between this histological group and adynamic bone disease. CONCLUSIONS: It is postulated that adynamic bone disease is a form of renal osteodystrophy, separate from osteomalacia, appearing when bone resistance to PTH develops, probably a transient stage to more hyperparathyroid histological classes with increasing severity of chronic renal failure.

Adolescent

Ultrastructure of the organic matrix of embryonic avian bone after en bloc reaction with various electron-dense 'stains'.

The morphological distribution of noncollagenous proteins in bone matrix is poorly known, chiefly because their amount changes with the type of bone, and they are not easily recognizable under the electron microscope. The aim of this study was to obtain further ultrastructural data on these proteins and on their relationships with collagen fibrils and the calcification process. Specimens of woven bone from the upper metaphysis of the tibia of chick embryos were examined after en bloc treatment with acetone solution of KMnO4, or aqueous or alcoholic solutions of phosphotungstic acid or uranyl acetate. Untreated specimens were used as controls. Untreated, decalcified, and decalcified and stained sections were examined. As expected, the results showed that the woven bone matrix of the chick embryo consists of irregularly oriented collagen fibrils which outline wide interfibrillary spaces. These contain calcification nodules at the calcification front and are full of irregularly oriented crystals in completely calcified matrix. These spaces contain greater than expected amounts of filamentous or granular organic structures (probably corresponding to noncollagenous proteins), which are in a close relationship with needle- and filament-like crystals. The collagen fibrils are calcified to a lower degree than the interfibrillary spaces, and often appear uncalcified, or decalcified, when the interfibrillary spaces do not. This suggests that either the former calcify after the latter, or the respective mineral phases differ in their degree of solubility, so that the decalcification of collagen fibrils precedes that of interfibrillary spaces. The results confirm that woven bone contains an amount of interfibrillar noncollagenous material greater than lamellar bone, and show that woven bone calcification occurs not only in and on collagen fibrils, but chiefly between them, in relationship with noncollagenous filamentous material.

Animals

The anatomy of bone sialoprotein immunoreactive sites in bone as revealed by combined ultrastructural histochemistry and immunohistochemistry.

Bone sialoprotein was immunolocalized at the EM level in thin Lowicryl K4M sections of rat bone. Because of the unconventional EM morphology of the bone matrix seen in thin demineralized acrylate sections, the pattern of immunolabeling was compared with detailed structural images of demineralized bone obtained using an en bloc treatment of tissue samples with the cationic electron 'dye,' Malachite Green (MG), which provides stabilization and retention of anionic material throughout specimen processing. A system of structures corresponding to the sites of bone sialoprotein (BSP) immunoreactivity, as seen in Lowicryl K4M this sections, could be readily identified in the MG-treated, epoxy thing sections. This system includes the cement lines, and aggregates of similar material within mineralized bone and mineralizing osteoid. The virtual identity of BSP distribution with the arrangement of the MG-visualized material indicates that a BSP-enriched, noncollagenous phase can be demonstrated using different, unrelated tissue preparation and imaging protocols for EM. Besides improving our understanding of the distribution of bone sialoprotein in bone, these data assign a previously unrecognized structural dimension to noncollagenous material in the bone matrix.

Animals

Prevention of ovariectomy osteopenia in rats after vaginal administration of Hyaff 11 microspheres containing salmon calcitonin.

The benzyl ester of hyaluronic acid (Hyaff 11) is a highly mucoadhesive polymer and can be processed into microspheres that can effectively deliver incorporated drugs by closely adhering to the mucosal surface and by protecting the drug from enzymatic inactivation. In this study, Hyaff 11 microspheres have been investigated as novel delivery system for the vaginal administration of salmon calcitonin (sCT) to ovariectomized rats. Moreover, this particular animal model has been used to evaluate the biological activity of the polypeptide after vaginal and I.M. administration by measuring the skeletal effect of sCT. A total of 66 female rats were used: 6 rats served as the basal group; 12 were subjected to sham operation and left without pharmacological treatment; 48 underwent bilateral ovariectomy and these were divided at random into four groups of 12 animals. One group was not submitted to pharmacological treatment; another was treated daily with sCT I.M.; in the other two groups, the hormone was incorporated into Hyaff 11 microspheres and was daily administered by vaginal route as dry powder or dispersed in pessaries. Sixty days after surgery, rats were sacrificed and the proximal third of the right tibia was removed and prepared for histomorphometry. Treatment with sCT, independent of the administration route, largely prevented the bone volume loss of the nontreated ovariectomized group. Prevention mainly depended on the maintenance of the trabecular number and on the increase of the trabecular thickness. Independent of the administration procedure, sCT greatly reduced the increase of tartrate-resistant acid phosphatase-positive poly- and, particularly, mononucleated osteoclasts found in the nontreated ovariectomized group.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Intravaginal

Extent of alkaline phosphatase cytochemistry vs. extent of tetracycline fluorescence in the evaluation of histodynamic variables of bone formation.

Alkaline phosphatase (ALP) activity is a new histomorphometric index of the extent of osteoblastic surfaces involved in mineralization. To assess its validity in the evaluation of bone formation, we carried out a comparative study between histomorphometric values obtained on the basis of the extent of tetracycline labeling and of the length of ALP-positive endosteal surfaces. The following variables were compared (indicated by ALP when based on the extent of ALP positivity): trabecular mineralizing surface (MS/BS vs. ALP.S/BS); osteoid mineralizing surface (MS/OS vs. ALP.S/OS); bone formation rate (BFR/BS vs. ALP.BFR/BS); and adjusted appositional rate (Aj.AR vs. ALP.Aj.AR). Bone biopsies from 39 patients with chronic renal failure and different types of renal osteodystrophy were considered (48 +/- 12 years of age; 19 men and 20 women). Patients were double labeled with tetracycline and biopsies were embedded in glycol-methacrylate at +4 degrees C. Patients showed various types of renal osteodystrophy and were assigned to different groups of pathologies. Although it differed in incidence according to the different groups, ALP activity was found in typical plump osteoblasts bordering osteoid seams and in flat cells, either in contact with osteoid or along the quiescent surfaces of bone in continuity with it. Tetracycline codistributed with all these features to variable extents, according to groups. In all patients, however, ALP.S/BS and ALP.S/OS respectively exceeded MS/BS and MS/OS. In consequence of this, ALP.BFR/BS and ALP.Aj.AR were greater than BFR/BS and Aj.AR, respectively. For each of the variable considered, differences among groups of patients with different types of renal osteodystrophy were highly significant. Good correlations were found between the variables measured with the two methods.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Ultrastructural organic-inorganic relationships in calcified tissues: cartilage and bone vs. enamel.

Close organic-inorganic relationships exist in all calcified tissues, the inorganic substance being linked to crystal ghosts (CGs). These are organic, crystal-like structures present in areas of initial calcification. In cartilage and bone, they form aggregates with the same morphology and distribution as the calcification nodules; in enamel, they consist of long filament- and ribbon-like structures, having the same arrangement as untreated crystals. CGs of cartilage and bone are acidic structures with histochemical properties of proteoglycans; CGs of enamel probably correspond to enamelins. The close morphologic similarity between CGs and crystals suggests that the former have a role in the formation of the latter.

Bone and Bones

Lectin binding properties of bovine resting cartilage.

The aim of this study was to evaluate the differential localisation of glycoconjugates of bovine hyaline cartilage matrix by lectin histochemistry, to compare the results of lectin histochemistry with those that can be obtained in the same tissue with PAS and alcian blue. Frozen and paraffin sections were stained with HE, PAS and alcian blue (pH 1.8). Alcian blue staining was carried out also after 1 and 24 hour digestion with bovine testicular hyaluronidase. Peroxidase conjugated WGA, PNA and RS lectins were tested on all sections before and after 1 hour digestion with bovine testicular hyaluronidase. The results show that all the lectins used in this study react with sugars linked to proteoglycans of territorial matrix, the reaction being increased in territorial, and induced in interterritorial matrix by 1 hour hyaluronidase digestion. Alcian blue at pH 1.8 and PAS were complementary, the former staining territorial, and the latter interterritorial matrix. After 1 hour hyaluronidase digestion, alcian blue stained also the interterritorial matrix. These results suggest that lectins react with low molecular weight proteoglycans and that short hyaluronidase digestion causes depolymerization of high molecular weight proteoglycans without loss of their glucidic components, allowing: a) penetration of alcian blue molecules into the macromolecular proteoglycan network; b) an increase of sugar residuals available for lectin histochemistry. Lectin histochemistry can be useful for differential localisation of glycoconjugates in bovine cartilage, especially if associated with short hyaluronidase digestion and conventional histochemical techniques.

Alcian Blue

Diagnostic value of serum peptides of collagen synthesis and degradation in dialysis renal osteodystrophy.

The assay of serum peptides of bone collagen formation and degradation could potentially provide an indirect estimate of the rate of bone turnover. In our study we have measured serum levels of the carboxy-terminal propeptide of type I procollagen (PICP) as a marker of bone formation and serum levels of the pyridinoline cross-linked telopeptide domain of type I collagen (ICTP) as a marker of bone resorption in 53 patients (47.7 +/- 10 years, M +/- SD) on haemodialysis (for 9.5 +/- 3.8 years) and affected by renal osteodystrophy. Besides PICP and ICTP, patients were also sampled for serum intact and C-terminal PTH, osteocalcin (BGP) and alkaline phosphatase (AP). A transiliac bone biopsy for histomorphometry was also performed in all. As expected both PTH assays, BGP and AP, were correlated reciprocally and to histomorphometric parameters. As for serum levels of PICP, they were on average increased (268.5 +/- 104.9 ng/ml, M +/- SD) compared to normals (range 66-176), but not correlated to classical humoral markers of hyperparathyroidism (PTH and AP), with the exception of BGP (with a rather low r value: 0.365, P < 0.01), nor to histomorphometric indices of bone resorption and formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase

Clinical features of 24 patients on regular hemodialysis treatment (RDT) for 16-23 years in a single unit.

Careful investigation of the clinical conditions of patients on maintenance hemodialysis for about 20 years in a single dialysis unit was of great interest for evaluation of the pathological consequences in long-term survivors of insufficient correction of uremia and of the dialysis treatment "per se". We analyzed the outcomes for a cohort of 116 patients who started RDT before 1976 and the clinical conditions of the 24 patients still on RDT in our unit at the end of 1991 (average duration of treatment = 222 +/- 23 months). Actuarial survival was 72% at 10 years and 43% at 20 years. Rehabilitation of the 24 survivors was rather good: 13 were able to work, 8 were retired or unable to work, but able to care for most personal needs. Actual body weight, anthropometric parameters and biochemical parameters revealed a well-preserved nutritional status. Anemia improved from 23 +/- 7 at the start of RDT to 31 +/- 8 in the 21 patients never treated with erythropoietin. Blood pressure was normal without therapy in 18 patients and elevated in 6. Mild-to-moderate left ventricular hypertrophy was present in all the 6 patients with arterial hypertension and in only 6 of the 18 normotensive patients. The ratio of early diastolic filling to filling during atrial contraction (E/A ratio) was < 1 in 16 patients: it was 1.05 +/- 0.43 in 9 patients with stable intradialysis blood pressure and significantly lower (0.73 +/- 0.15) in 12 patients with recurrent intradialysis hypotension. Supraventricular arrhythmias were detected by Holter monitoring in 41% and ventricular arrhythmias in 35% of patients. Extensive vascular calcifications were present (in 100% of patients in the abdominal aorta), but only 4 patients showed clinical signs of peripheral vascular disease. Subperiosteal resorption was detected radiologically in the hands of 59% of patients. Bone histology, interpretable for only 20 patients, revealed no bone lesions in 1 case (5%), mild mixed osteodystrophy in 3 cases (15%), advanced mixed osteodystrophy in 5 cases (25%), osteodystrophy with predominant hyperparathyroidism in 2 cases (10%), osteodystrophy with predominant osteomalacia in 6 cases (30%), and aplastic bone disease in 3 cases (15%). Moderate aluminum staining was found in only 4 patients and was more marked in earlier biopsies taken before withdrawal of the aluminium-containing phosphate-binding drugs.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent

Metabolic acidosis and osteodystrophic bone disease in predialysis chronic renal failure: effect of calcitriol treatment.

The role of metabolic acidosis on osteodystrophic bone lesions of chronic renal failure has been studied retrospectively in 24 patients, divided into two equal groups of 12, one with normal acid-base equilibrium (group A) and one with metabolic acidosis (group B). The two groups were found to differ significantly in serum levels of BGP (23.7 +/- 18 vs. 42.3 +/- 24 ng/ml, p < 0.02) and in several bone histomorphometric parameters such as osteoid volume (4.5 +/- 3.4 vs. 10.2 +/- 6.6%, p < 0.01), osteoid surface (27.7 +/- 18 vs. 48.4 +/- 19%, p < 0.01), single-labelled surface (7.94 +/- 2.9 vs. 15.8 +/- 9.9%, p < 0.02), mineralizing surface (60.69 +/- 26 vs. 30.89 +/- 15.8%, p < 0.003) and mineralization lag time (56.5 +/- 54 vs. 170.5 +/- 189 days, p < 0.05), with the acidotic group showing excess osteoid and a defect in mineralization. Osteomalacia was found only in the acidotic group, while the only 2 cases of adynamic bone disease (ABD) were in the nonacidotic group. Calcitriol administration, 0.25 micrograms daily for a period of 1 year, in 5 cases in group A and 6 cases in group B induced significant improvement of bone lesions mainly in group A. Two of these patients following treatment acquired the characteristics of ABD. In group B, the response to treatment was very limited, with 5 patients still showing persistence of the histological mixed type of bone disease. In conclusion, metabolic acidosis is accompanied by osteomalacia, pure or mixed variety, and shows a relative resistance to calcitriol administration. Normal acid-base equilibrium is more frequently associated with mild hyperparathyroidism and ABD, spontaneously or as a consequence of calcitriol administration.

Acid-Base Equilibrium

Morphological studies of hypomineralized enamel of rat pups on calcium-deficient diet, and of its changes after return to normal diet.

BACKGROUND: Micro-hardness investigations have shown that rat pups nursed by mothers on a low calcium diet and weaned with the maternal calcium-deficient diet develop hypomineralized enamel. The inorganic and organic components of this enamel, their relationships, and their changes after return to normal diet have been studied by light and electron microscopy. METHODS: The maturation zone of incisor enamel has been studied in: (1) rats nursed for 20 days by mothers on a low calcium diet and weaned for 30 days with the same diet (E1 enamel); (2) rats that after the calcium-deficient diet were fed normal diet for 10 days (E2 enamel); and (3) rats nursed for 20 days by mothers on a normal diet and weaned for 30 days with a normal diet (controls). RESULTS: The results showed that E1 enamel was hypomineralized, as noted by its Azure II-Methylene blue stainability in undecalcified sections, its light staining with the von Kossa method, and its ultrastructure. E1 crystallites, although present throughout the whole enamel, were thinner than those of E2 enamel, which were similar to those of controls. E1 interrod crystallites were thicker in the intermediate than in the dentinal zone and were thicker than rod crystallites. Organic matrix was present throughout the whole E1 enamel. Its organic components (crystal ghosts) had the same shape, arrangement, and organization as those of inorganic crystallites. Crystal ghosts were greatly reduced in E2 enamel and in controls. CONCLUSIONS: The results lead to the conclusions that: (1) E1 enamel is hypomineralized, and its degree of calcification is restored by return to a normal calcium diet; (2) intra- and interprismatic calcification occurs in a different way; (3) crystallite thickness is initially greater in dentinal than in the superficial zone and is reversed as crystallite growth is completed; and (4) loss of enamel proteins is necessary for completion of crystallite growth and not for crystallite formation.

Animal Nutritional Physiological Phenomena

Evidence of healing of secondary hyperparathyroidism in chronically hemodialyzed uremic patients treated with long-term intravenous calcitriol.

The aim of this study was to assess the effect of a long-term course of high-dose i.v. pulses of calcitriol (CLT) on hyperparathyroid bone disease (HBD) and functional mass of parathyroid glands of chronically hemodialyzed uremic (CHU) patients. We prospectively studied nine CHU patients treated with CLT, 30 ng/kg/body wt, i.v., thrice weekly over a period of eight months. Plasma concentrations of intact parathyroid hormone (iPTH), bone GLA protein (bGLA) and bone isoenzyme of alkaline phosphatase (biALP) were sampled throughout. Transiliac bone biopsies were made before and after the start of CLT therapy. Double scanning scintigraphy of the neck with 201Tl-99Tc was made before, during and eight months after the start of the treatment. All patients but one, who later responded to higher than planned CLT doses, had significant decreases of plasma iPTH (F = 76; P < 0.0001; ANOVA). The mean pretreatment value of PTH was 966 +/- 160 (mean +/- SE) pg/ml and it had decreased significantly by the first week (T = 2.4, P < 0.04), and had fallen an average of 80% by the 35th week. Ionized plasma calcium concentration was 1.19 +/- .01 mmol/liter which rose significantly (F = 13.5; P < 0.0001) by the 14th week to maximal peak levels, averaging 1.34 +/- .02 mmol/liter. Changes in biALP were parallel to those of iPTH, while bGLA tended to increase immediately after the start of the therapy and to significantly decrease thereafter (T = 3.2; P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Intravenous versus oral calcitriol therapy in renal osteodystrophy: results of a prospective, pulsed and dose-comparable study.

Intravenous calcitriol is generally considered to be more efficient than oral administration in the treatment of secondary hyperparathyroidism of chronic renal failure, although a comparative and prospective study employing the same doses and modality of drug administration is lacking. We therefore evaluated 12 hemodialysis (HD) patients (51.7 +/- 9.4 years, mean +/- SD, HD for 8.7 +/- 4.7 years) with marked secondary hyperparathyroidism. Based on basal humoral and bone histologic parameters, we divided these patients into 2 comparable groups. Calcitriol (0.015 micrograms/kg) was given at the end of each dialysis intravenously in group A and orally in group B. Humoral parameters were evaluated basally and after 1, 2, 4 and 8 months. Ax bone biopsy was taken at the start and at the end of the study. From the first month of treatment, group A showed an increment in ionized calcium (from 1.28 +/- 0.08 to 1.37 +/- 0.12 mmol/l, p < 0.01), with a reduction in intact parathyroid hormone (from 470.1 +/- 349.5 to 255.5 +/- 256.5 pg/ml; p < 0.0003) and alkaline phosphatase (from 615.1 +/- 696.3 to 445.3 +/- 577.7 mU/ml, p < 0.001). The occurrence of hypercalcemia prompted a reduction in dialysate calcium content in 4 of 6 patients after 4 months, and of the calcitriol dose in 2 of 4 patients after 6 months. Ionized calcium then turned to 1.32 +/- 0.11 (p = n.s. compared to basal) while the intact parathyroid hormone concentration tended to revert (363.3 +/- 360 pg/ml, p = n.s. compared to basal) and alkaline phosphatase remained low (420 +/- 638 mU/ml, p < 0.0005).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral

Culture and differentiation of chondrocytes entrapped in alginate gels.

We studied the response to culture conditions and the differentiative ability in suspension culture in alginate gels of resting chondrocytes from the preosseous cartilage of adult pig scapula. It was found that the maximum rate of chondrocyte duplication is reached at the fourth day in culture whereas the rate of proteoglycan synthesis and alkaline phosphatase expression do not gain a maximum value before the seventh day. During the culture time, the chondrocytes undergo differentiation as it is demonstrated by the alkaline phosphatase specific activity increase and by morphological criteria (hypertrophy, increase of the number of mitochondria per cell, increased endoplasmic reticulum, matrix vesicle production). The alginate gels can be easily dissolved to obtain cell populations in which the variation of cytosolic calcium concentration following a proliferative stimulus can be conveniently observed using the conventional procedure of Fura 2.

Alginates

Long-term survival on renal replacement therapy for primary hyperoxaluria type I.

We describe the case of a patient in end-stage renal failure due to primary hyperoxaluria type I (PH1) who started hemodialysis in 1977 and is still alive and active. The diagnosis of PH1 was first suspected after a bone biopsy performed in 1981 to investigate hyperparathyroidism. Oxalosis recurred as early as 3 months after transplantation in a cadaver kidney grafted in 1987; nevertheless, graft function remained good enough to make possible the discontinuation of dialysis treatment for 5 months and thereafter to have only 1 dialysis a week for 17 months. The diagnosis of PH1 has been recently confirmed despite the patient being already anuric by means of the determination of plasma oxalate and glycolate levels as well as by determining hepatic alanine:glyoxylate amino-transferase.

Adult