Biomedical subjects

P Dubois

Publications and source records attributed to P Dubois.

At least 19 recordsLinked to original sources

PubMed2001

Cold-adapted beta-galactosidase from the Antarctic psychrophile Pseudoalteromonas haloplanktis.

The beta-galactosidase from the Antarctic gram-negative bacterium Pseudoalteromonas haloplanktis TAE 79 was purified to homogeneity. The nucleotide sequence and the NH(2)-terminal amino acid sequence of the purified enzyme indicate that the beta-galactosidase subunit is composed of 1,038 amino acids with a calculated M(r) of 118,068. This beta-galactosidase shares structural properties with Escherichia coli beta-galactosidase (comparable subunit mass, 51% amino sequence identity, conservation of amino acid residues involved in catalysis, similar optimal pH value, and requirement for divalent metal ions) but is characterized by a higher catalytic efficiency on synthetic and natural substrates and by a shift of apparent optimum activity toward low temperatures and lower thermal stability. The enzyme also differs by a higher pI (7.8) and by specific thermodynamic activation parameters. P. haloplanktis beta-galactosidase was expressed in E. coli, and the recombinant enzyme displays properties identical to those of the wild-type enzyme. Heat-induced unfolding monitored by intrinsic fluorescence spectroscopy showed lower melting point values for both P. haloplanktis wild-type and recombinant beta-galactosidase compared to the mesophilic enzyme. Assays of lactose hydrolysis in milk demonstrate that P. haloplanktis beta-galactosidase can outperform the current commercial beta-galactosidase from Kluyveromyces marxianus var. lactis, suggesting that the cold-adapted beta-galactosidase could be used to hydrolyze lactose in dairy products processed in refrigerated plants.

Adaptation, Physiological↗

PubMed2000

Ultrastructure of sea urchin calcified tissues after high-pressure freezing and freeze substitution.

The improvements brought by high-pressure freezing/freeze substitution fixation methods to the ultrastructural preservation of echinoderm mineralized tissues are investigated in developing pedicellariae and teeth of the echinoid Paracentrotus lividus. Three freeze substitution (FS) protocols were tested: one in the presence of osmium tetroxide, one in the presence of uranyl acetate, and the last in the presence of gallic acid. FS in the presence of osmium tetroxide significantly improved cell ultrastructure preservation and should especially be used for ultrastructural studies involving vesicles and the Golgi apparatus. With all protocols, multivesicular bodies, suggested to contain Ca(2+), were evident for the first time in skeleton-forming cells. FS in the presence of gallic acid allowed us to confirm the structured and insoluble character of a part of the organic matrix of mineralization in the calcification sites of the tooth, an observation which modifies the current understanding of biomineralization control in echinoderms.

Animals↗

PubMed2000

Mimicking a conformational B cell epitope of the heat shock protein PfHsp70-1 antigen of Plasmodium falciparum using a multiple antigenic peptide.

The Pf72/Hsp70-1 antigen is a major target in the naturally acquired immunity against Plasmodium falciparum malaria. We carried out an extensive analysis of the responses to several epitopes on the least conserved C-terminal domain, according to the mode of sensitization: malaria infection or immunization with different immunogens. We found significant differences in the panel of B-cell epitopes recognized by animal models including primates, and by humans sensitized by natural infection. We focused the analysis on one epitope that is unique to Plasmodium species. It is specifically recognized by a monoclonal antibody that mediates the killing of infected hepatocytes in vitro. We produced a polymeric multiple antigenic peptide (MAP) form of this sequence, which enabled us to identify a new B-cell epitope not detected by ELISA with linear peptides. The polymer was strongly recognized by sera from monkeys or humans sensitized by natural infection, whereas the monomer was not. We modelled the three-dimensional structure of the Pf72/Hsp70-1 sequence, using known Escherischia coli DnaK structures as a template. This predicted that the corresponding region would form a loop in the native antigen. The results presented here suggest that the MAP strategy is also particularly useful as a means of obtaining suitable synthetic models for conformation-dependent epitopes.

Amino Acid Sequence↗

PubMed2000

In vivo comparison between computed tomography and magnetic resonance image analysis of the distal radius in the assessment of osteoporosis.

In a prospective case-control cross-sectional study, we investigated the usefulness of both computed tomography (CT) and magnetic resonance (MR) image analysis of the distal radius for distinguishing a small sample of fractured osteoporotic women from control women regardless of bone mineral density. The study population included 12 subjects who were divided into two groups according to their bone status. The first group consisted of six women with at least one vertebral fracture occurring in the absence of high-energy trauma, and the second group consisted of six women without disease affecting bone mass or bone metabolism. Cross-sectional and coronal slices were obtained from both CT and MR systems. For CT images, the slice thickness was 1000 jim and the plane resolution was approx 200 jim x 200 jim. MR images were obtained from a 1.5-T imager with a two-dimensional spin-echo Ti-weighted sequence leading to a slice thickness of 2000 jim and a plane resolution of 195 jim x 195 jim. Bone texture analysis was performed using fractal and structural methods leading to the measurement of 23 features. Most of the structural variables derived from histomorphometric parameters and were measured after segmentation from a binary or a skeletonized image. Bone densitometry was measured by dual-energy X-ray absorptiometry both at the lumbar spine and the nondominant femoral neck. On cross-sectional slices, 12 variables, mainly obtained from structural analysis, were significantly different between the two groups for CT images (p < 0.05) against two variables only for MR images (p < 0.05). The number of variables statistically different between the two groups was significantly higher for CT images compared with MR images (p = 0.003). In the same way, odds ratios for fracture per 1 control group standard deviation decrease were significant for 10 variables on CT images, whereas, in contrast, none of the variables measured on MRI images led to significant odds ratios. The results obtained for the two methods on coronal slices were poorer without a difference between either CT or MR images in terms of discrimination between fracture and nonfracture subjects. In conclusion, this study suggests that bone texture analysis obtained from CT compared with MRI offers a best discrimination between controls and osteoporotic patients, probably the result to the spatial resolution. which is better for CT than for MR images.

Absorptiometry, Photon↗

PubMed2000

Does high-resolution computed tomography image analysis of the distal radius provide information independent of bone mass?

This study aimed to investigate the usefulness of computed tomography (CT) image analysis of the distal radius for comparing two groups of postmenopausal women matched for age and bone mineral density at both the lumbar spine and femoral neck. The first one consisted of 16 women with at least one vertebral fracture and the second consisted also of 16 women without disease affecting bone mass or bone metabolism. Eight slices were selected in each patient: four consecutive coronal slices and four consecutive axial slices. Bone texture analysis was performed using structural methods leading to the measurement of 24 features. Most of the structural variables derived from histomorphometric parameters and were measured after segmentation from a binary or a skeletonized image. Nine variables were significantly different between the two groups on axial slices: valley number, valley surface area, apparent bone volume/tissue volume (BV/TV), apparent trabecular separation, apparent trabecular number, trabecular bone pattern factor, trabecular skeletal length, node count, and node-to-node strut count. Also four variables were significantly different between osteoporotic women and controls on coronal slices: apparent BV/TV, trabecular partition, node-to-node strut count, and terminus-to-terminus strut count. In conclusion this study suggests that bone texture analysis could yield additional data on bone mass for explaining bone strength and therefore could be used for improving the prediction of fracture risk.

Absorptiometry, Photon↗

PubMed1999

Cellular and antibody responses to the Plasmodium falciparum heat shock protein Pf72/HSP70 during and after acute malaria in individuals from an endemic area of Brazil.

Proliferative and antibody responses to three synthetic peptides corresponding to Pf72/ HSP70 were followed-up in acute malaria patients from an endemic area of Brazil. In vitro lymphocyte responsiveness to all peptides was relatively low and short-lived and there was a considerable variation in the frequency and magnitude of the individual lymphoproliferative response to the peptides at different periods after the onset of infection. Although 96% of the patients had IgG antibodies to crude Plasmodium falciparum asexual blood stage antigens, specific IgG antibody responses to the peptides varied from 12.5 to 40% according to the tested peptides. No significant difference was observed in the proliferative or antibody responses to the peptides between individuals that remained parasitemic after treatment and those that recovered from malaria infection. The different frequencies of proliferative responses in peripheral blood T cells on different occasions after the onset of their infection show that, in order to be informative, evaluation of the in vitro cellular immune response to peptides requires longitudinal studies in which each individual is tested repeatedly.

Acute Disease↗

PubMed1999

Image analysis of the distal radius trabecular network using computed tomography.

Bone texture analysis might provide information about bone structure in a noninvasive manner. In a prospective case-control cross-sectional study we investigated the value of computed tomography (CT) image analysis of the distal radius in the assessment of osteoporosis. Twenty patients suffering from postmenopausal osteoporosis were studied and compared with 21 age-matched controls. Eight slices were selected in each patient: four consecutive coronal slices and four consecutive cross-sectional slices. Bone texture analysis was performed using statistical, fractal and structural methods leading to the measurement of 32 features. Structural variables derived from histomorphometric parameters were measured after segmentation from a binary or a skeletonized image. Bone mineral density was measured by dual-energy X-ray absorptiometry both at the lumbar spine and the femoral neck. Eight of the 9 statistical features were significantly different in osteoporotic women as compared with controls (coronal slices, p < 0. 05). Seven structural variables were statistically different between the two groups on coronal slices (p < 0.05): valley surface area, bone volume/tissue volume, trabecular partition, Euler's number, trabecular bone pattern factor, node-to-node strut count and terminus-to-terminus strut count. The most significant results on coronal slices (p < 0.01) concerned 4 structural features: trabecular partition, Euler's number, trabecular bone pattern factor and terminus-to-terminus strut count. Three features were statistically different (p < 0.01) between the two groups on cross-sectional slices (skeletonization from gray levels). A few features yielded by texture analysis were correlated with both lumbar spine and femoral neck bone mineral density, but the level of these correlations was weak (r < 0.5). In conclusion, CT image analysis of the distal radius is a useful tool for characterizing bone texture alterations in osteoporotic women. These findings are in keeping with microarchitectural osteoporosis-related changes diagnosed on bone biopsies.

Aged↗

PubMed1999

Direct qualitative and quantitative characterization of a radiosensitizer, 5-iodo-2'-deoxyuridine within biodegradable polymeric microspheres by FT-Raman spectroscopy.

Non-destructive qualitative and quantitative characterization of a radiosensitizer, 5-iodo-2'-deoxyuridine (IdUrd), incorporated within injectable microspheres of a biodegradable polymer, poly(D,L-lactide-co-glycolide) (PLGA), was performed using Fourier transform (FT) Raman spectroscopy. Raman spectra of IdUrd, free and entrapped in microspheres, were recorded under fluorescence-free conditions, described and assigned. For the Raman bands of the PLGA microspheres, assignments with preferential localization of the corresponding vibrations at lactic or glycolic units were proposed. No evidence for drug-polymer interactions in microspheres was found. This allowed the FT-Raman spectra to be used for the quantification of the IdUrd content in the samples. For the microspheres with IdUrd loadings varying from 2 to 27% of the total weight, the methodology used provided good reproducibility and precision (1%). Within the sensitivity of the technique, samples exposed to sterilization doses (27 kGy) of gamma-radiation did not exhibit marked changes in the drug structure.

Humans↗

PubMed1999

Ultrastructural localization of proteins involved in sea urchin biomineralization.

Three skeletal tissues of the adult echinoid Paracentrotus lividus (the pedicellaria primordium, the test, and the tooth) were immunolabeled with three sera raised against the total mineralization organic matrix and two specific matrix proteins (SM30 and SM50) from the embryo of the echinoid Strongylocentrotus purpuratus. Two conventional chemical fixation protocols and two high-pressure freezing/freeze-substitution protocols were tested. One conventional protocol is recommended for its good preservation of the ultrastructure, and one high-pressure freezing/freeze-substitution protocol is recommended for its good retention of antigenicity. Immunolabeling was obtained in the three adult tissues. It was confined to the active skeleton-forming cells and to the structured organic matrix. The results indicate that the matrix proteins follow the classical routes of secretory protein assembly and export and suggest that SM30 and SM50 are a part of the tridimensional network formed by the organic matrix before the onset of mineralization. They show that the genetic program of part of skeletogenesis is conserved among different calcification models and developmental stages.

Animals↗

PubMed1999

Particle counting immunoassay for urinary cotinine. Comparison with chromatography, enzyme-linked immunoassay and fluorescence polarization immunoassay.

Urinary cotinine was measured according to its inhibitory activity on the agglutination of cotinine-coated latex particles by anti-cotinine antibodies, the agglutination being measured by optical counting of the remaining non-agglutinated particles (particle counting, PaC). The detection limit was 0.03 microgram/ml and the practical range extended from 0.03 to 3.9 micrograms/ml. The correlation results of 320 urine samples with those of high pressure liquid chromatography, enzyme-linked (Coti-Tracq EIA, Serex Inc., Maywood, NJ, USA), and fluorescence polarization immunoassay (TDX instrument, Abbott, Abbott Park, IL, USA) were r = 0.90, r = 0.69, r = 0.87, respectively, whereas the correlation coefficients between the assays other than particle counting ranged from 0.62 to 0.88. PaC does not require any separation step and can thus be easily automated.

Chromatography, High Pressure Liquid↗

PubMed1999

Serial measurement of calcaneal pitch during midstance.

Although emerging evidence suggests a causal relationship between arch structure and musculoskeletal injury, few investigations have adequately assessed arch function during gait. In this study, digitized videofluoroscopy was used to evaluate the sagittal plane motion of the calcaneus during gait. Nine female subjects requiring diagnostic foot radiographs underwent videofluoroscopy. The calcaneal inclination angle, arch height ratio, and tarsal index were digitally analyzed for all radiographic images. Calcaneal pitch was correlated to both the arch height ratio and the tarsal index. Repeated measures analysis of variance helped to identify a significant reduction in the mean calcaneal pitch during the midstance and early propulsive periods of gait. These findings suggest that although calcaneal pitch may be used as an indicator of rearfoot position, biomechanical classification of foot types based on radiographs may result in erroneous conclusions concerning foot function.

Adult↗

PubMed1998

Virtual reality versus conventional training in retinal photocoagulation: a first clinical assessment.

Virtual reality can provide a new method for apprenticeship in surgery. We have developed a training simulator for retinal photocoagulation. This paper describes a comparison between this new training tool and the conventional teaching method. Residents were divided into two groups, each group being trained exclusively with one of the methods. A final evaluation was made by a single expert, who did not know which training mode the residents had undergone. The results show that this new training mode is at least as efficient as the current one and may reduce training duration. While experts remain in control of the contents of training and assessment of skill, this study outlines new prospects for apprenticeship and evaluation in the field of medicosurgical techniques. In addition, this preliminary study emphasizes the difficulties in introducing a new modality in a traditional teaching environment.

Humans↗

PubMed1998

Ultrastructure and cytochemistry of the early calcification site and of its mineralization organic matrix in Paracentrotus lividus (Echinodermata: Echinoidea).

The ultrastructure and cytochemistry of skeleton formation sites prior to mineralization are described for the first time in echinoderms. These early sites are intracellular vacuoles located in syncytial pseudopodia of skeleton-forming cells. They contain a mineralization organic matrix, which shows a calcium-binding ability and is framed in a tridimensional structure made of concentric layers bridged by radial threads. This organic matrix presents repetitive structures which could be implicated in mineralization control. Both the tridimensional organization of the organic matrix and its framing, before mineralization starts, question the current theories which suggest that the echinoderm organic matrix is soluble at the onset of mineralization and adsorbs on the forming crystal.

Animals↗

PubMed1998

Regional insertional mutagenesis on chromosome III of Arabidopsis thaliana using the maize Ac element.

The authors describe the production and characterization of a collection of Arabidopsis lines each carrying a transposed Ac (trAc) element. A total of 507 lines were obtained following germinal transpositions of a single Ac element located on the upper portion of chromosome III. Southern analysis revealed that up to 90% of the lines in this collection harbour distinct insertions of Ac in the Arabidopsis genome. As previous studies on the behaviour of Ac in Arabidopsis have indicated that approximately two out of three transposition events occur to linked loci, the authors hypothesized that this collection could be of great use in isolating insertional mutants for genes located in the vicinity of the donor locus. PCR and phenotypic screens were performed to identify mutations in five loci located within a 40 cm region of chromosome III centered on the donor locus. Molecular analyses confirmed the presence of germinal insertions of Ac in three of the loci (NPTII, ABI3 and EST #210A22). At a fourth locus (AtDMC1), despite the absence of a germinal insertion, one line in which somatic insertions occurred regularly was identified and may be of use in isolating a germinal insertion. This collection of trAc lines constitutes a useful complement to the existing collection of T-DNA insertion lines and will soon be made available through the Arabidopsis Biological Resource Center.

Arabidopsis↗

PubMed1998

Asthma and airway hyperresponsiveness among Belgian conscripts, 1978-91.

BACKGROUND: Although there is convincing evidence that the prevalence of asthma among children has increased over the last three decades, it remains uncertain whether such an upward trend has occurred in adults. The aim of this study was to assess whether the prevalence of asthma has changed in young Belgian adults in recent years. METHODS: A retrospective analysis was conducted of available statistics from the Belgian Armed Forces for the period 1978-91 because conscripts who reported a history of past or current asthma at call-up examination underwent standardised assessment of non-specific airway responsiveness by military chest physicians. Exemption from military service due to asthma was strictly based on the objective evidence of airway hyperresponsiveness. RESULTS: A mean of 48,331 conscripts aged 17-31 years were examined annually from 1978 to 1991. The prevalence of reported asthma rose from 2.4% in 1978 to 7.2% in 1991, while the proportion of asthmatics with airway hyperresponsiveness remained little changed at 48.4% in 1978 and 51.4% in 1991. CONCLUSIONS: The observed increase in prevalence of reported asthma was not accompanied by a decrease in the proportion of conscripts with objectively measured airway hyperresponsiveness. These observations provide supporting evidence that the increase in asthma symptoms was not simply due to reporting bias.

Adolescent↗

PubMed1998

Sagittal plane motion of the human arch during gait: a videofluoroscopic analysis.

Despite an abundance of literature investigating arch structure and musculoskeletal injury, there seems to be little consensus regarding the most appropriate technique of measuring dynamic arch motion. In this study, digitized videofluoroscopy was used to determine the sagittal plane motion of the medial longitudinal arch during dynamic gait. Nine female subjects requiring diagnostic foot radiographs underwent videofluoroscopy during a normal gait cycle. The calcaneal inclination angle, calcaneal-first metatarsal angle (CI-MT1) and height-to-length ratio of the arch, all reputed to measure arch alignment, were digitally analyzed from static radiographic images. Both the calcaneal inclination angle (0.96) and CI-MT1 (-0.98) angles were highly correlated with the criterion measure of height-to-length ratio. Repeated measures analysis of variance (ANOVA) identified a significant increase in the mean CI-MT1 angle during stance, suggesting a continual lowering and elongation of the arch. This study questions the validity of characterizing foot motion based on static measures of arch shape and recommends that further research be conducted to establish whether the observed trends reflect normal or pathological foot function.

Adult↗

PubMed1998

CT scan texture analysis of the distal radius: influence of age and menopausal status.

OBJECTIVES: To gain information on bone architecture by performing bone texture analysis in a sample of women covering a broad age range. PATIENTS AND METHODS: We studied 29 healthy women aged 23 to 80 years (55 +/- 18 years), 19 of whom were postmenopausal. None was taking drugs known to influence bone mass or bone metabolism. Computed tomography of the nondominant distal radius was performed with 1 mm slice thickness and 1 mm gap. Four consecutive coronal and four consecutive axial sections were selected for each patient and entered into a PC-type computer. Bone texture was evaluated using grey level run length analysis (five parameters), differential local variation analysis (four parameters), fractal analysis (two parameters), trabecular network extraction and three-dimensional relief characterization. The mean of each study parameter for the four coronal sections and for the four axial sections was calculated. Absorptiometry was done in 16 patients. RESULTS: Linear correlations with age were strongest (P < 0.001) for parameters measured on coronal sections by trabecular network extraction, i.e., trabecular bone volume (r = -0.68), trabecular plate separation (r = -0.65), total skeletal length (r = -0.71), number of nodes (r = 0.73), number of node-node segments (r = -0.74) and trabecular bone pattern factor (r = 0.71). Also, these parameters were significantly different between premenopausal women (33 +/- 9 years) and postmenopausal women (67.3 +/- 9 years). Correlations between bone mineral density and texture parameters were few in number and modest in strength, suggesting that the parameters measured may reflect bone structure rather than bone mass. CONCLUSION: Bone texture in women undergoes changes with advancing age that may reflect alterations in bone microarchitecture.

Adult↗

PubMed1998

A clinical decision analysis of cryotherapy compared with expectant management for cervical dysplasia.

BACKGROUND: Screening for precursors of cervical cancer with colposcopic examination for women with abnormal Papanicolaou (Pap) smears identifies those with cervical dysplasia. Though the majority of mild dysplasias (CIN I) will regress, many are treated with cryotherapy. METHODS: We used decision analysis to compare immediate cryotherapy with expectant management (following with another Pap smear or colposcopy, with treatment reserved for progression or a duration of 2 years). The decision tree included the possibility of more invasive surgical procedures if the cryotherapy was ineffective or if the dysplasia progressed in extent of cervical involvement or in grade. Probabilities were derived from literature review and expert judgment. The analysis considered the disutility of the follow-up examinations, cryotherapy, and the more invasive procedures, using expert assessment. RESULTS: Using the baseline assumptions, expectant management led to a better outcome for most patients (57%), who recover with no procedure. However, more patients treated with expectant management required surgical procedures (loop electrosurgical excisional procedure, conization, or, rarely, hysterectomy) than did those treated with immediate cryotherapy. In the expected disutility analysis, expectant management was better than immediate cryotherapy. Sensitivity analysis showed that three factors had the potential to change the recommendation of the analysis: (1) the probability the dysplasia will regress, (2) the disutility of the process of expectant management, and (3) the disutility of invasive procedures compared with cryotherapy. CONCLUSIONS: The analysis indicated that expectant management is preferable to immediate cryotherapy for women with histologically proven mild cervical dysplasia. However, this conclusion depended on assumptions about three factors for which there is insufficient evidence in the literature. More research is needed.

Colposcopy↗