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Biomedical subjects

J Aubia

Publications and source records attributed to J Aubia.

At least 19 recordsLinked to original sources

Osteoblasts in HIV-infected patients: HIV-1 infection and cell function.

BACKGROUND: HIV-infected patients have been shown to have a severe alteration in osteoblast function that appears to be related to the infection. OBJECTIVE: To determine whether normal human osteoblasts express CD4, whether osteoblasts from patients with HIV infection are infected by HIV-1 and whether osteoblast dysfunction observed in vivo also occurs in vitro. METHODS: Osteoblast cultures from bone marrow biopsies of HIV-infected patients (n = 14) and control patients (n = 10) were used in a cross-sectional study and a case-control prospective study. Expression of CD4 was analysed using flow cytometry and reverse transcriptase polymerase chain reaction; the presence of HIV-1 particles was determined by measuring p24 antigen in the supernatants of osteoblast cultures and viral DNA or RNA in the osteoblasts using the polymerase chain reaction. Osteoblast function was assessed by measuring cell proliferation, type I collagen and osteocalcin synthesis. RESULTS: In human osteoblasts, CD4 expression could not be determined using flow cytometry, although low levels of mRNA coding for CD4 were detected. HIV infection was not observed in osteoblast cultures from HIV-infected patients nor was there any alteration in replication and synthesis of type I collagen, although osteocalcin synthesis was increased. CONCLUSIONS: It is unlikely that HIV-1 infects human osteoblasts in vivo; therefore, the hypothesis that these cells could act as local HIV-1 reservoirs should be reconsidered.

Adult↗

In vitro synthesis of type I collagen: quantification of carboxyterminal propeptide of procollagen type I versus tritiated proline incorporation.

Radioimmunoassay of the carboxyterminal propeptide of human type I procollagen has been recently introduced to measure in vitro synthesis of type I collagen by osteoblasts and fibroblasts. However, it has not been assessed whether the equivalent results are obtained with this new assay and with tritiated proline incorporation to collagen protein. To this purpose, both methods were used to quantify synthesis of type I collagen in normal human osteoblast cultures to which fetal calf serum and human serum were added in order to stimulate protein synthesis. A positive correlation in the results obtained by both methods was obtained (r = 0.95, P = 0.0001). Given the technical advantages of detection of levels to carboxyterminal propeptide of type I procollagen, we consider that this is the technique of choice for the quantification of in vitro synthesis of type I collagen by normal human osteoblasts.

Aged↗

DNA image cytometry. An alternative method in osteoblast proliferation assays.

OBJECTIVE: To assess whether DNA image cytometry can be used as an alternative method to tritiated thymidine uptake quantification in osteoblast proliferation assays. STUD DESIGN: Proliferation of normal human osteoblasts incubated with normal human serum at 0%, 2.5%, 5%, 10%, 20% and 40% was quantified by tritiated thymidine uptake quantification and DNA image cytometry. RESULTS: Tritiated thymidine uptake quantification showed that normal human serum stimulated the proliferation of normal human osteoblasts and that the degree of stimulation was directly related to the concentration of serum in the culture medium. Similar results were obtained when the DNA image cytometry assay was used. A highly significant linear relationship between the ranks of both methods was found (Spearman's r = 1.00, P = .0253). CONCLUSION: DNA image cytometry may be a valuable alternative when the use of radioactive material is not desired and/or subsequent morphologic or immunocytochemical characterization of cells under study is required.

Aged↗

Immunoreactive osteocalcin forms in conditioned media from human osteoblast culture and in sera from healthy adult control subjects and patients with bone pathologies.

BACKGROUND: The aim of this work was to study the immunoreactive forms of bone Gla protein (BGP) present in conditioned media of human osteoblast cultures (BGP released from osteoblast) and in the sera of healthy adult control subjects and patients with bone pathologies (chronic renal failure on haemodialysis, Paget's disease of bone and post-menopausal osteoporosis). METHODS: The technical procedure used was a combination of high-performance liquid chromatography (HPLC) and different BGP assays with several specificities to analyse BGP levels in the different HPLC fractions. Aliquots of conditioned media or sera were purified through a Sephadex G-50m column and by HPLC (C4 reverse-phase column) in a 25-40% acetonitrile gradient. Two-minute fractions were collected and divided into three aliquots in order to determine osteocalcin content using three different assays: (a) ELSA-OST-NAT IRMA, which only detects intact osteocalcin; (b) ELSA-OSTEO IRMA, which detects intact osteocalcin and N-terminal fragments; and (c) OSCA Test RIA, which detects intact osteocalcin, C-terminal and other fragments. RESULTS: We found different immunoreactive forms of osteocalcin in the culture medium of human osteoblasts and in sera from control subjects and patients for the bone pathologies studied. We did not find great qualitative differences between the immunoreactive osteocalcin profile found in the culture medium from human osteoblasts and the sera from healthy control subjects. However, the different bone pathologies show different characteristic patterns of immunoreactive forms of osteocalcin. CONCLUSIONS: An interesting finding has been the detection, both in sera and in osteoblast culture media, of several immunoreactive forms of intact osteocalcin that eluted from HPLC at different acetonitrile percentages, and therefore correspond to different molecular forms.

Adult↗

99mTc-labelled red blood cell scintigraphy for localization of gastrointestinal bleeding in chronic renal failure.

Angiodysplasia (AD) may be the source of bleeding in patients with gastrointestinal hemorrhage, with special occurrence in the elderly population and in patients with chronic renal failure (CRF). Although several techniques have been tested for its diagnosis, the gold standard is not well defined yet. We analyze the usefulness of 99mTc-labelled red blood cell (99mTc RBC) scintigraphy in the localization of bleeding from AD lesions in a cohort of 21 patients. Other investigative methods include fibrocolonoscopy examination, angiography, or diagnostic laparotomy. Group A (AD and CRF): 11 patients. Group B (AD without CRF): 10 patients. 99mTc RBC scintigraphy showed 88.9% sensitivity and specificity in group A, while in group B it had 100% sensitivity and specificity. Arteriography showed 100% sensitivity and specificity. On the contrary, fibrocolonoscopy had a very low sensitivity (30%). Our results suggest that 99mTc RBC scintigraphy may be the preferred diagnostic tool for AD, especially in patients with CRF, in whom arteriography may accelerate the decline of renal function.

Aged↗

Acute effects of ethanol on mineral metabolism and trabecular bone in Sprague-Dawley rats.

In order to assess the effects of acute ethanol intoxication on bone, 45 female Sprague-Dawley rats were studied. Five rats were sacrificed at baseline. The remainder received either ethanol (2 g/kg of body weight) intraperitoneally or isotonic saline. Rats were sacrificed in groups of 10 (5 intoxicated and 5 placebo) at 1, 4, 8, and 24 hours after injection. At the time of sacrifice, a blood sample was obtained and the 4th vertebra was excised for histomorphometric analysis of undecalcified bone. Effect of ethanol was assessed by an analysis of variance test using a Scheffé procedure. In ethanol-treated rats we observed (mean +/- SD, ethanol versus controls, maximum difference point, P value) a significant decrease in osteiod surface with osteoblasts (42.86 +/- 15.61% versus 64.57 +/- 6.24%, P < 0.05); osteoclast number (0.05 +/- 0.02 n/mm2 versus 0.17 +/- 0.09 n/nm2, P < 0.05), and osteocalcin (36.9 +/- 2.21 ng/ml versus 45.8 +/- 5.1 ng/ml, P < 0.05). Osteoclast surface was initially reduced (0.129 +/- 0.09% versus 0.425 +/- 0. 26%, P < 0.01) but showed a subsequent increase (0.765 +/- 0.24% versus 0.226 +/- 0.17%, P < 0.01) attributable to alcohol. There was also a significant decrease in serum Ca (8.51 +/- 0.23 mg/dl versus 9.10 +/- 0.29 mg/dl, P < 0.01) and parathyroid hormone values (23.51 +/- 5.72 pg/ml versus 76.39 +/- 11.66 pg/ml, P < 0.001). We conclude that acute alcohol intoxication in rats induces early striking changes in bone histology and analytical parameters, not completely reversed after 24 hours. These data are consistent with a toxic effect induced by alcohol on bone.

Acid Phosphatase↗

Osteoblastic proliferation in bone biopsies from patients with end-stage chronic renal failure.

Osteoblasts have traditionally been considered to be terminally differentiated cells and therefore unable to divide. Data in recent years, however, indicate that cellular differentiation does not usually preclude preservation of proliferative ability and that most differentiated cells are able to divide under adequate stimuli. The aim of this study was to assess whether cubic osteoblasts undergo proliferation during the formation phase of the remodeling cycle under a stimulus that increased bone turnover. For that purpose, the osteoblastic proliferation index (OPI) was analyzed by DNA image cytometry in transiliac bone biopsies from 33 patients with chronic renal failure (23 men, 10 women; mean age 50.4 +/- 15.1 years) who have been classified into low (n = 13), normal (n = 15), and high (n = 15) bone turnover according to activation frequency (Ac.f). OPI was significantly higher (p < 0.002) in the high bone turnover group (13.90 +/- 4.72%) compared with the low (2.38 +/- 4.13%) and normal turnover groups (2.84 +/- 4.04%). There was a positive correlation between OPI and the following histomorphometric parameters: bone formation rate, surface referent (r = 0.76, p = 0.00001), activation frequency (r = 0.73, p = 0.00001), mineral apposition rate (r = 0.73, p = 0.00001), bone formation rate, volume referent (r = 0.71, p = 0.00001), and mineralizing surface (r = 0.62, p = 0.0001). This study shows that a rise in bone turnover is associated with a marked increase of bone-forming cell proliferation in patients with end-stage chronic renal failure. From this finding, it may be concluded that cubic osteoblasts do not behave as "terminally differentiated" cells in vivo, because a high proportion of them are still able to divide.

Adult↗

Bone remodeling alterations in myelodysplastic syndrome.

There is a close relationship between hematopoietic bone marrow and bone cells. Thus, the profound derangement of hematopoiesis in myelodysplastic syndromes (MDS) might be expected to affect bone cell function. We studied the dynamic histomorphometric changes in bone in 22 MDS patients to examine this relationship and analyze the influence of hematological disease on bone remodeling. Bone-regulating hormones and histomorphometry of undecalcified transiliac bone biopsies, after double tetracycline labeling, were studied. Serum calcium, phosphorus, creatinine, alkaline phophatase, osteocalcin, iPTH, 25(OH)D3, 1,25(OH)2D3, hydroxyprolinuria, and calcium/creatinine ratio in urine were normal compared with controls. Histomorphometry showed a significant decrease in osteoblast surface (Ob.S/BS) (0.30 +/- 0.40 vs. 0.8 +/- 1.1, p = 0.031), wall thickness (W.Th), (22.03 +/- 5.5 vs. 31.8 +/- 5.8, p < 0.005), osteoclast number (N.Oc/T.Ar) (0.004 +/- 0.01 vs. 0.017 +/- 0.01, p = 0.03), mineral apposition rate (MAR) (0.16 +/- 0.15 vs. 0.53 +/- 0.19, p < 0.005), bone formation rate, surface referent (BFR/BS) (0.004 +/- 0.10 vs. 0.016 +/- 0.016, p = 0.009), and activation frequency (Ac.f) (0.06 +/- 0.07 vs. 0.21 +/- 0.23, p = 0.008). An increase in mineralization lag time (MLT) (119.2 +/- 78.6 vs. 29.6 +/- 77, p < 0.005), (mean +/- SD, unpaired Student t-test) was observed. Bone volume (BV/ TV), eroded surfaces (ES/BS), and osteoid thickness (O.Th) remained unchanged. This picture of adynamic bone with decreased mineral apposition rate and markedly decreased osteoclast number is a characteristic finding in MDS patients. Thus, bone histomorphometric finding in MDS patients show the relationships and interactions between hematopoietic and bone cells.

Aged↗

Bone remodelling in human immunodeficiency virus-1-infected patients. A histomorphometric study.

The aim of this study was to identify and describe possible alterations of bone histomorphometry in patients with human immunodeficiency virus (HIV-1) infection and to assess the relation between these alterations and disease severity. Forty-four HIV-1-infected patients seen successively at our hospital were evaluated for the study. In an attempt to avoid confounding factors as far as possible, we excluded patients who fulfilled any of the following criteria: age less than 18 or greater than 40 years; recent history of extended bed rest; previous diagnosis of metabolic bone disease, renal insufficiency, or hepatic failure; clinical or echographic signs of liver cirrhosis; diabetes mellitus or previous diagnosis of other endocrine diseases; drug therapy that could act on bone metabolism; and/or moderate to severe nutritional alteration. Twenty-two patients (13 men, 9 women; age: 27.9 +/- 4.1 years, mean +/- standard deviation) were included in the study. Plasma and urine biochemistry and calcium-regulating hormones were determined. Bone mineral content was measured on vertebrae L2 to L4 and on the neck and intertrochanteric areas of the femur by dual-photon absorptiometry. A transiliac bone biopsy was performed after double-tetracycline labelling, with histomorphometric study of undecalcified bone. Serum osteocalcin was found to be lower in patients who, according to the Centers for Disease Control (CDC) classification, had greater disease severity, and showed a positive correlation with the number of CD4+ T lymphocytes. No alterations in bone densitometry were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorptiometry, Photon↗

Bone growth and modeling changes induced by periosteal stripping in the rat.

In this study, the changes in longitudinal bone growth and metaphyseal modeling induced by middiaphyseal periosteal stripping in the rat femur were analyzed by means of histomorphometrical techniques. One hundred forty-four male 30-day-old Sprague-Dawley albino rats distributed in 4 groups of 36 were studied: a control group, a sham group, a group with middiaphyseal right femoral periosteal stripping, and a group with a polyethylene ring wrapped around the stripped zone. The animals were euthanized at 1, 2, or 4 weeks from the start of the experiment, after double tetracycline labeling. A statistically significant, albeit small, longitudinal overgrowth of stripped femurs was observed after a latency period of 2 to 4 weeks. The metaphyseal diameters were greater in stripped femurs than nonstripped femurs. This finding was associated with a lower osteoclastic index in the external metaphyseal surface and with a lower bone formation rate in the internal surface of the metaphyseal cortex. These latter findings have not been reported previously in the literature and may support the role of the periosteum in controlling metaphyseal modeling.

Animals↗

Effect of cyclosporine A on normal human osteoblasts in vitro.

UNLABELLED: Cyclosporin-A (CsA) inhibits in vitro proliferation of non-human tumour-cloned osteoblasts. Our aims were to study the direct effect of CsA on proliferation of normal human osteoblast (NHOb) cultures and to ascertain whether CsA-treated patients' sera (CsATPS) may exert effects on the osteoblast which differ from the direct effects of CsA. We studied tritiated thymidine ([3H]thymidine) incorporation in NHOb cultures incubated with (a) increasing CsA concentrations (1.2 to 4800 ng/ml), (b) the same concentrations as in the previous experiment but with the addition of 20% fetal calf serum (FCS) or 20% normal human serum (NHS), (c) 40% NHS or 40% CsATPS. Results at 96 h in (a) CsA inhibited uptake from 300 ng/ml, in (b) CsA inhibited [3H]thymidine uptake from 2400 ng/ml for cultures with FCS and 4800 ng/ml for cultures with NHS, in (c) CsATPS produced [3H]thymidine uptake inhibition compared with NHS. CONCLUSION: CsA alone inhibited [3H]thymidine incorporation in NHOb from concentrations similar to therapeutic concentrations. With FCS or NHS, inhibition was produced at higher concentrations. CsATPS inhibited at CsA concentrations lower than those of the two previous experiments.

Blood↗

Correlation between densitometric and hystomorphometric values in isolated vertebrae of Sprague-Dawley rats.

The study of bone mass in experimental animals usually requires invasive techniques. Dual energy X-ray absorptiometry (DEXA) may be an alternative as a non-invasive method (1). Bone mineral density (BMD) and bone mineral content (BMC) of 62 vertebrae of Sprague Dawley rats (SDr) measured by DEXA densitometry were compared with histomorphometric bone volume measurements, and a statistically significant correlation was found (r = 0.79 and 0.75, respectively, p < 0.001). In conclusion, DEXA is an accurate and feasible technique for the study of trabecular bone mass in SDr.

Absorptiometry, Photon↗

Long-standing remission after 25-OH D3 treatment in a case of chronic myelomonocytic leukaemia.

This report describes a case of chronic myelomonocytic leukaemia (CMML) in whom a complete remission was achieved and sustained 15 months after treatment with 25-OH vitamin D3. No side-effects were observed. Although vitamin D1 has been used in the treatment of myelodysplastic syndromes, to our knowledge this is the first case of long-standing remission in a patient with CMML. This 'differentiation therapy' might be considered as a possible alternative in the management of this disease.

Aged↗

Bladder involvement in systemic amyloidosis causing massive hemorrhage in hemodialysis.

We report a chronic hemodialyzed patient with bladder involvement of a secondary amyloidosis that presented as isolated hematuria evolving quickly to a massive hemorrhage and vesical rupture. We believe that this is the first report of bladder amyloidosis involvement in the course of hemodialysis. This knowledge may help in managing dialysis patients with hematuria.

Aged↗