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Biomedical subjects

L Moro

Publications and source records attributed to L Moro.

At least 55 records · Page 3Linked to original sources

Clinical performances of galactosyl hydroxylysine, pyridinoline, and deoxypyridinoline in postmenopausal osteoporosis.

We have previously shown that galactosyl hydroxylysine (GHYL), pyridinoline (PYD), and deoxypyridinoline (DPD) have a better accuracy and discriminate power than hydroxyproline in distinguishing postmenopausal osteoporotic women from premenopausal controls. In this study, we evaluated the clinical performances of GHYL, PYD, and DPD, alone or in combination, in distinguishing postmenopausal osteoporotic women (OPBD, n = 26) from age-matched controls (CBD, n = 19). The diagnosis of osteoporosis was based upon the bone density (BD) of the lumbar spine measured by quantitative computed tomography (CBD: BD > 108 mg/cm3; OPBD: BD < 70 mg/cm3). Urinary excretion of GHYL, PYD, and DPD were measured by HPLC, and all data were expressed as the molar ratio with the creatinine excretion (GHYL/CR, PYD/CR, and DPD/CR). The clinical performances were tested by: Z score analysis (Z), Receiver Operated Characteristic curve analysis (%Acc) and logistic-regression analysis of the posterior probabilities for prediction from a logistic model (LOGIST). GHYL/CR, PYD/CR, and DPD/CR were significantly increased in OPBD compared with CBD. The clinical performances were similar for the three assays, with slightly better performances for GHYL/CR (GHYL/CR: Z = 3.14, %Acc = 70 +/- 8, LOGIST P = 0.01; PYD/CR: Z = 2.19, %Acc = 67 +/- 8, LOGIST P = 0.051; DPD/CR: Z = 2.13, %Acc = 65 +/- 8, LOGIST P = 0.06). None of the possible combinations of the three assays yielded better clinical performances than GHYL/CR alone. In conclusion, this study further confirms the validity of GHYL, PYD, and DPD as markers of bone resorption.

Aged↗

Genomic organisation and chromosomal localisation of the gene encoding human beta adducin.

Adducin (ADD) is a heterodimeric protein of the membrane skeleton with subunits of 103 (alpha) and 97 kDa (beta). It promotes the assembly of the spectrin-actin network. We have previously shown that one point mutation in each of the alpha and beta rat ADD-encoding genes is associated with blood pressure variation in an animal model for hypertension, the Milan hypertensive strain of rats, probably due to a change in the phosphorylation pattern. In fact, the rat mutations, Y to F for alpha and R to Q for beta, are located, respectively, in a tyrosine kinase and a protein kinase A phosphorylation site. We have now determined, for the human beta-ADD-encoding gene, its chromosomal localisation, exon-intron organisation and alternative splicing patterns. We report here that human beta-ADD is localised on chromosome 2 and we also show a characteristic 3' end alternative splicing of the beta-ADD RNA that generates two distinct beta-ADD families, namely ADD 63 and 97; both of them in turn present a very complex differential splicing pattern in the internal exons.

Alternative Splicing↗

Galactosyl hydroxylysine in assessment of Paget's bone disease.

Urinary galactosyl hydroxylysine/creatinine ratio (GHL) was used to assess rates of bone collagen degradation and the activity of the pagetic lesion as well as for monitoring the rate and degree of suppression of bone resorption over 1 year in patients treated with 30 mg of intravenous pamidronate for 3 consecutive days. The clinical utility of GHL was compared with that of urinary hydroxyproline/creatinine and deoxypyridinoline/creatinine and with bone isoenzyme of serum alkaline phosphatase. The results suggest that GHL is a quantitative marker of the activity of Paget's bone disease. GHL is less sensitive than hydroxyproline, deoxypyridinolone and bone alkaline phosphatase in monitoring treatment of Paget's disease. The assay of GHL is easier, faster and less costly than that of hydroxyproline or deoxypyridinoline and it can be easily standardized.

Adult↗

Detection of pyridinium cross-links in human bile.

This study evaluated whether pyridinium cross-links, which are positively charged, besides renal clearance are also cleared by the liver into bile. In 13 human bile samples tested, we were able to detect both pyridinoline (PYD) and deoxypyridinoline (DPD) in small amounts which were estimated to be about 1-2% of the amount usually found in urine. To further evaluate the amount of pyridinium cross-links excreted through bile, we studied the stability of these compounds at the alkaline pH of bile. No effect on their stability was detected over a 6-hour incubation. The origin of these molecules in bile and the significance of this finding in the use of PYD and DPD as bone resorption markers are discussed.

Adult↗

Biochemical markers of bone metabolism in the assessment of osteoporosis.

Several biochemical markers are available for the study of bone metabolism. The characteristics of these markers are reviewed and their potential application in osteoporosis is examined. It is concluded that biochemical markers are potentially useful in identifying those at risk of developing osteoporosis, selecting appropriate treatment, monitoring therapy, and studying the pathogenesis of osteoporosis.

Animals↗

Angiokeratoma corporis diffusum and arteriovenous fistulas with dominant transmission in the absence of metabolic disorders.

BACKGROUND: A three-generation family with members affected by angiokeratoma corporis diffusum (ACD) and arteriovenous fistulas of the legs is described. Our purpose was to investigate possible lysosomal storage defects previously described in association with ACD. OBJECTIVE: Results of physical examination of both affected and unaffected family members were otherwise normal as was the life span. The inheritance pattern of both ACD and arteriovenous fistula traits was autosomal dominant, with variable expressivity and incomplete penetrance. Microscopic examination of ACD lesions showed dilated capillaries without vacuolation of cells. Ultrastructural studies failed to reveal lysosomal abnormalities. Normal levels of alpha-galactosidase, beta-galactosidase, alpha-fucosidase, and alpha-sialidase were detected in peripheral blood leukocytes and skin fibroblasts. CONCLUSIONS: The association of autosomal dominant ACD and arteriovenous fistulas might represent a novel syndrome. However, pathogenesis of these lesions remains unknown.

Adolescent↗

Time-relationship between bone growth and increment of bone mineral content in growing rats.

The aim of the study was to describe the time-relationship between the growth rate of the rat tibia and the increase rate of its mineral content. Appositional and endochondral bone growth rates were derived from sequential X-rays measurements of tibial widening and lengthening, respectively; the increase rate of bone mineral content was derived from sequential photon absorptiometry measurements of the proximal tibio-fibular site. The time-relationship of appositional growth rate and endochondral growth rate versus bone mineral content increase rate was mathematically described. The results allow a better understanding of the time-course of two distinct features of bone growth: increase in size and increase in mineral content.

Absorptiometry, Photon↗

Correction of the defective extracellular matrix of Ehlers-Danlos syndrome skin fibroblasts by dexamethasone.

Skin fibroblasts derived from Ehlers-Danlos syndrome (EDS) patients lack an organized extracellular matrix (ECM) of fibronectin (FN) and often show an accumulation of cytoplasmic FN. The treatment of EDS cells of different types (I to VIII) with 10(-7) M dexamethasone (dex), as well as cocultivation with control fibroblasts, induced in most cases the assembly of a FN-like ECM. The study of FN mRNA expression by dot-blot hybridization and of FN released into the culture media of EDS cells showed that the correction of the defective FN-ECM of EDS cells by dex treatment is associated in most cases with an increase of FN mRNA synthesis and of secreted FN.

Cells, Cultured↗

The natural history of pre-type 1 (insulin-dependent) diabetes mellitus in patients with autoimmune endocrine diseases.

An 11-year prospective study was carried out in 180 non-diabetic patients with organ-specific autoimmune diseases to evaluate islet cell antibodies in predicting Type 1 (insulin-dependent) diabetes mellitus. Islet cell antibodies were characterised according to titres, persistence, complement-fixing ability, and pattern. During follow-up, 14 of 46 patients with islet cell antibodies persistently greater than 5 Juvenile Diabetes Foundation Units (JDF-U) (30.4%), none of 23 with islet cell antibodies between 2.5 and 5 JDF-U or fluctuating, and 3 of 109 without islet cell antibodies (2.7%), developed diabetes. The cumulative risk of developing diabetes was 70%, 0%, and 4%, respectively. All the patients who developed diabetes were females. Eight progressed to insulin-dependence acutely, four showed a transient period of non-insulin-dependence, while two were still insulin-free. No difference was found in titres of islet cell antibodies for the risk of diabetes. Complement-fixing islet cell antibodies enhanced the cumulative risk for the disease in patients with conventional islet cell antibodies at low-middle (> or = 2.5-40 JDF-U), but not at high (> or = 80 JDF-U) titres. Forty-two patients with islet cell antibodies were investigated for the whole or the selective pattern. In the presence of the whole pattern the cumulative risk for diabetes rose to 100%, while with the selective pattern it declined to 34%. The whole pattern was found in 83% of patients who developed Type 1 diabetes acutely. In patients with organ-specific autoimmune diseases, the whole islet cell antibody pattern greatly enhances the prediction for diabetes.

Adolescent↗

Effect of bisphosphonate therapy and parathyroidectomy on the urinary excretion of galactosylhydroxylysine in primary hyperparathyroidism.

BACKGROUND: In patients with mild or asymptomatic primary hyperparathyroidism a reliable index of bone resorption might be useful for appropriate management. Hydroxyproline is the most commonly used marker of bone resorption but its low specificity and sensitivity are known. Galactosylhydroxylysine, an amino acid mainly represented in bone collagen, has been proposed as a more suitable index of bone resorption. In this study we evaluated the sensitivity of galactosylhydroxylysine and hydroxyproline assays in following the changes of their urinary levels in 12 patients with mild primary hyperparathyroidism before and after treatment with bisphosphonate and surgery. METHODS: Serum and fasting urine specimens were obtained from 12 women with mild primary hyperparathyroidism before and after bisphosphonate treatment (2.5 mg daily for 5 days, intravenously) and after a further 25 days; in 7 patients biochemical tests were also performed 1 and 6 days after parathyroidectomy. Galactosylhydroxylysine was assayed by an HPLC method and hydroxyproline by a RIA commercial kit. RESULTS: Baseline galactosylhydroxylysine urinary levels were far above the normal range in all the patients whilst in 8 of them baseline hydroxyproline levels were normal. Bisphosphonate treatment significantly decreased bone turnover as shown by a significant fall in serum calcium (from 2.9 to 2.6 mmol/l; P < 0.001) and in galactosylhydroxylysine and hydroxyproline (-55 and -31% respectively). Twenty-five days after the end of treatment, resorption increased again and serum calcium and galactosylhydroxylysine, but not hydroxyproline, rose significantly towards basal levels. One day after parathyroidectomy serum calcium, galactosylhydroxylysine and PTH showed reduction below normal ranges. PTH and galactosylhydroxylysine returned to normal values at day 6 after parathyroidectomy. No changes in hydroxyproline levels were seen. Galactosylhydroxylysine, but not hydroxyproline, correlated significantly with serum calcium and PTH. CONCLUSION: Galactosylhydroxylysine appears to be a sensitive index of bone resorption, useful in the clinical assessment of bone involvement and in the management of patients with mild primary hyperparathyroidism.

Aged↗

["Prophylactic" video-laparoscopic cholecystectomy].

The authors examine the prophylaxis of infections caused by cholelithiasis in kidney and heart transplant candidates as a new indication for videolaparoscopic cholecystectomy (VLC). The study included 6 patients in dialysis for chronic renal insufficiency and one patient suffering from cyanogenic congenital cardiopathy with asymptomatic gallbladder calculosis. The results obtained show that there are no substantial differences compared to patients without associated pathologies and justifies the inclusion of "prophylactic" VLC in preparatory treatment protocols for kidney and heart transplant in patients suffering from cholelithiasis. The authors emphasise the necessary technical measures to prevent hemorrhage, intraoperative loss of CO2 and postoperative laparoceles.

Adult↗

[Transrectal polypectomy using a TEM (transanal endoscopic microsurgery) technique].

The paper describes the removal of a broad-based tubulo-villous adenoma from the middle rectum using a transanal endoscopic microsurgical technique. This procedure, which was introduced by Buess, Thess and Hutterer in 1983, represents an alternative to the classic transanal technique, compared to which it is more accurate, and to the trans-sacral and transperitoneal methods compared to which it is less invasive.

Aged↗

High-performance liquid chromatographic and mass spectrometric identification of phosphocitrate synthesized by human kidney homogenate.

Two high-performance liquid chromatographic techniques for the analysis of phosphocitrate, a metabolite synthesized by the cell that is thought to play a role in a number of physiological as well as pathological events, have been developed. The mass spectra of the isolated compound obtained under fast-atom bombardment and collisionally activated decomposition mass-analysed ion kinetic energy conditions are also reported.

Chromatography, High Pressure Liquid↗

Rat cortical and trabecular bone collagen glycosylation are differently influenced by ovariectomy.

This study sought to evaluate whether estrogen depletion influences the hydroxylysine glycosydes content of rat bone collagen. For this reason thirty 100 day old female rats were divided in 6 groups of 5 rats each. Three groups were ovariectomized and 3 groups were sham-operated. The animals were sacrificed at 115, 130 and 145 days of age (i.e., 15, 30 and 45 days after surgery). Cortical and trabecular bone was prepared from tibiae and femurs. Hydroxylysine glycosydes content was measured by HPLC. Ovariectomy is followed, in the rat, by an increased hydroxylysine glycosylation in trabecular bone but by a constant or slightly decreased hydroxylysine glycosylation in the cortical bone. In view of the different effects of estrogens on the two bone compartments previously reported, a possible functional explanation of these findings is proposed.

Animals↗

The glycosides of hydroxylysine are final products of collagen degradation in humans.

Glucosyl-galactosyl-hydroxylysine (GGHYL) and galactosyl-hydroxylysine (GHYL) are metabolites derived from collagen degradation. They are useful biochemical markers provided they are not further processed. Experiments were run to test the activity of alpha- and beta-glycosidases in human kidney cortex preparations. Results allow to exclude the presence of the specific enzymes, in contrast with what is reported for the rat kidney tissue.

Animals↗

Quantitative evaluation of mRNAs by in situ hybridization and image analysis: principles and applications.

We describe an image analysis (IA) program that has been developed for the quantitative evaluation of mRNA evidenced by in situ hybridization (ISH) with radiolabeled probes in cultured cells and tissue sections. ISH-IA allowed the detection and quantitative evaluation of mRNA expressed by heterogeneous in vitro cultured cells. This method, when combined with dot-blot analysis, allowed the evaluation of the approximate number of mRNA molecules expressed by single cells. IA permitted the evaluation of cultured cells' morphological parameters (such as cytoplasm and nucleus areas) modifications in relation to specific mRNAs expression, which can vary during cell cycle, development, aging, and in different pathologies and treatment with drugs. ISH-IA was applied for the evaluation of mRNA isoforms generated by alternative splicing in single cells. This methodology was also applied for the semiquantitative evaluation and comparison of mRNA levels expressed by different cell types in human normal and tumor tissue sections.

Alternative Splicing↗

Galactosyl hydroxylysine and deoxypyridinoline: a methodological comparison.

Galactosyl hydroxylysine and deoxypyridinoline are at present the most promising markers of bone resorption. Various studies have indeed shown that these two markers discriminate with high accuracy subjects with different rates of bone turnover and that their accuracies and discriminate power are very similar. The aim of this paper is to compare the practicality and the reproducibility of the HPLC galactosyl hydroxylysine and deoxypyridinoline assays. In summary, this review shows that the galactosyl hydroxylysine and deoxypyridinoline HPLC assays differ mainly in the need, in using deoxypyridinoline, for an acid hydrolysis and a preextraction of the urine samples. This implies two major problems for deoxypyridinoline: 1) more time is required due to the cumbersome preanalytical procedures; and 2) a lower reproducibility. Our data, in fact, show that both the intra-assay and inter-assay coefficient of variation of the deoxypyridinoline assay are almost 100% higher than those of the galactosyl hydroxylysine assay.

Adult↗