Biomedical subjects
Publications and source records attributed to C Roy.
The authors report an unusual case of retroperitoneal mucinous cystadenoma in contact with the isthmus of a horseshoe kidney and associated with congenital absence of the left ovary. The retroperitoneal site is very unusual, as only about twenty cases have been reported in this site. The pathogenesis of these tumours remains unclear and the authors discuss the various hypotheses proposed to date.
Trypanosoma cruzi and Toxoplasma gondii are protozoan parasites capable of causing infections of the nervous system. In order to determine effects of infection by these organisms on intercellular communication in the brain, dye coupling and connexin abundance and distribution were examined in leptomeningeal cells and astrocytes infected with T. cruzi or T. gondii. For both cell types infected with either type of protozoan parasite, intercellular diffusion of intracellularly injected Lucifer Yellow was dramatically reduced. Immunocytochemistry with antibodies specific for connexin43 (in astrocytes) or both connexin43 and connexin26 (for leptomeningeal cells) demonstrated that punctate gap junctional staining was much reduced in infected cells, although uninfected neighbors could display normal connexin abundance and distribution. Western blot analyses revealed that connexin43 abundance in both cell types infected with either parasite was similar to that in uninfected cells. Phosphorylation state of connexin43 (inferred from electrophoretic mobility of connexin43 isoforms) was not significantly affected by the infection process. Immunocytochemistry of whole brains from animals acutely infected with either parasite also showed a marked reduction in connexin43 expression. We conclude that infection of both types of brain cells with either protozoan parasite results in a loss of intercellular communication and organized gap junction plaques without affecting expression levels or posttranslational processing of gap junction proteins. Presumably, these changes in gap junction distribution result from altered targeting of the junctional protein to the plasma membrane, and/or from changes in assembly of subunits into functional channels.
OBJECTIVE: Previously, we showed that 15-20% of patients with rheumatoid arthritis (RA) have oligoclonal expansions of peripheral blood CD8+ T cells expressing T cell receptors encoded by the V(alpha)12 (AV12S1) gene. To better understand the significance of these expansions, the present study was undertaken to determine their specificity. METHODS: We cloned and characterized V(alpha)12+,CD8+ T cells from the peripheral blood of 1 RA patient with a clonal expansion of these T cells. RESULTS: The T cell clones were autoreactive since they recognized autologous, but not allogeneic, antigen-presenting cells. Upon activation, these T cells secreted interleukin-4 and interleukin-10. The autoreactive T cell clones were class I major histocompatibility complex (MHC) restricted, by either HLA-B60 or HLA-Cw3. CONCLUSION: A large population of class I MHC-restricted CD8+ T cells in a patient with RA is clonally expanded and autoreactive. These cells define a novel immune aberration in RA and provide a tool for defining the autoantigens that activate expanded T cell populations in vivo.
Three unusual cases of small-size leiomyosarcoma of the perirenal space were studied with CT. The renal capsule has been proved to be the origin of this type of tumor. A CT examination is accurate in suggesting the site of origin and excluding a renal cell carcinoma. However, unless evidence of invasion is noted, it is impossible on CT features to discriminate leiomyosarcoma from a benign leiomyoma.
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Laser energy is able to ablate, coagulate, and vaporize tissues. Its transmissibility in thin optical fibers makes it an ideal tool for use in percutaneous procedures. This article describes two applications in interventional musculoskeletal radiology. In percutaneous laser disc decompression the laser source is used to vaporize a small portion of the nucleus pulposus. In interstitial laser photocoagulation of osteoid osteoma the laser energy is used to coagulate and destroy the tumor by direct heating.
PURPOSE: To evaluate the clinical and biologic safety of ultrasmall superparamagnetic iron oxide particles (AMI-227) as a contrast agent for magnetic resonance (MR) lymphography and to assess their efficacy for the differentiation of metastatic and benign nodes in patients with urologic and pelvic cancer. MATERIALS AND METHODS: Thirty adults suspected of having lymph node metastases underwent MR imaging before and 22-26 hours after intravenous infusion of AMI-227 (1.7 mg Fe/kg). Sixty histopathologically proved lymph nodes were analyzed on MR images, and 29 of these nodes were also analyzed quantitatively. RESULTS: AMI-227 was well tolerated with no major side effects. It allowed the detection of 10 additional nodes relative to those detected at MR imaging without AMI-227. None of the 27 metastatic nodes showed a decrease in signal intensity (SI) on AMI-227-enhanced images; nine of 27 metastatic nodes showed an increase in SI on T1-weighted images, probably resulting from altered capillary permeability in the tumor. A visually perceptible reduction in SI, indicating active AMI-227 uptake, was observed on postcontrast T2- and T2*-weighted images in 16 of 21 benign nodes. The SI ratio of benign nodes was lower than that of metastatic nodes on T2- and T2*-weighted images. The sensitivity of AMI-227-enhanced MR lymphography was 100%, and the specificity was 80%. CONCLUSION: AMI-227 is safe and may facilitate the differentiation of metastatic and benign nodes in patients with urologic and pelvic cancers.
A very unusual CT appearance of renal cel carcinoma is presented, in which the fatty density mimicked a benign angiomyolipoma.
UNLABELLED: THE PURPOSE OF THE STUDY was to evaluate interstitial laser photocoagulation for local destruction of osteoid osteoma, with computed tomographic (CT) guidance. MATERIAL AND METHODS: 28 patients (age range from 5 to 48 years) with presumed osteoid osteoma were treated with CT-guided interstitial laser photocoagulation of the nidus. A high power semiconductor diode laser (805 nm) with a 400 microns optical fiber was used. The fiber was introduced into the nidus through a 18-gauge needle. Around the fiber tip, well-defined coagulative necroses from 5 to 9 mm (energy delivery, 400-1000J) were obtained. RESULTS: 27 patients had complete pain relief, which was effective within 24 hours in 18 patients. One patient had pain recurrence after 6 weeks. The remaining nidus was treated secondarily with complete relief. Treatment was unsuccessful in one patient, and surgical excision was performed. All patients were followed up for more than 1 year, with no sign of recurrence. The only notable complication was a mild reflex sympathetic dystrophy of the wrist in one patient. Sclerosis of the nidus was observed 6-12 months after the procedure. CONCLUSION: Percutaneous interstitial laser photocoagulation of osteoid osteoma seems to be a promising, simple, precise, and minimally invasive alternative to traditional surgical and percutaneous ablations.
The ultraviolet radiation (UVR) exposures of primary school children in Brisbane, Toowoomba and Mackay (latitudes 27 degrees 30', 27 degrees 33' and 21 degrees 15' south, respectively) were assessed over a period of 2 weeks at each location using UVR-sensitive polysulfone (PS) film badges attached at the shoulder. The students filled in questionnaires on their time spent outdoors for each day of the study. These data in conjunction with the ambient UVR measured by a detector/datalogger unit at each site were used to correlate the calculated exposures with those measured using the PS badges. Overall, the questionnaires indicated that the males spent more time outdoors and had higher measured UVR exposures than females. For both boys and girls at each location, there was a strong correlation between the mean measured UVR exposure and the ambient solar UVR at that location.
In this work the imagery of non-small cell primary bronchial cancers are reviewed. The standard chest x-ray of the thorax remains irreplaceable for the early detection of these pathologies. On the other hand, progress in imagery has propelled thoracic computed tomographic scanning to the pole position for assessing the local and regional extension of the disease as well as for distance spread of these cancers. The latest developments with spiral CT have again improved the performance of CT scanning. Magnetic resonance imaging has a few precisely defined roles in assessing the extension of bronchopulmonary cancers. In particular with involvement of the apices, the chest wall and in the extension into the cardiovascular system. In this article the emphasis is on the illustration of non-small cell bronchial cancers on computed tomography and on magnetic resonance imaging.
The present study evaluates the impact of an early stimulation program for 24 mother-infant (0-6 months) vulnerable dyads from the Montérégie region. These dyads participate in a series of ten stimulation workshops. Twenty four other dyads form a witness group. The program's major objective aims at developing positive interactions for the mother and stimulative interactions for the infant. The hypotheses state that at the end of the program, the mother from the intervention group will feel less isolated, possess a higher level of knowledge on the development of their child, perceive more positively the temperament of their child and have behaviors that are more contingent to their infants as well as more functional to this development. The two first hypotheses are not confirmed. However, at the end of the program, the mothers of the intervention group perceive the temperament of their child as foreseeable. Also, they imitate more frequently the verbal and facial behaviors of their infant and emit more vocalizations when they are interacting with the child. The discussion of results bears on the importance of modifying conditions in which the parent-child interactions are held to prevent deficits in the development of the children.
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We have cloned and ligated a full-length bovine link protein (LP) in the pMAL-c2 vector and overexpressed it in fusion with maltose-binding protein (MBP) in Escherichia coli. We have demonstrated dose-dependent binding of MBP/LP to biotinylated hyaluronan in a dot blot assay. A greater percentage of the expressed fusion protein was soluble, monomeric, and undegraded when the growth temperature was lowered, the growth medium was supplemented with zinc, and metal chelators were omitted from the lysis buffers. Similar effects were observed when we tested the effects of lower growth temperature and zinc supplementation on another construct consisting of MBP in fusion with the first proteoglycan tandem repeat of LP. Our results suggest zinc may be necessary for the folding and disulfide bond formation of recombinant LP. In addition, a greater amount of monomeric MBP/LP produced at 27 degrees C with zinc supplementation bound to biotinylated hyaluronic acid-binding region of aggrecan than MBP/LP produced at 27 or 37 degrees C without zinc. This suggests that recombinant LP may have a conformational requirement for zinc necessary for binding to aggrecan. Factor Xa cleavage of MBP/LP expressed in the presence of zinc yielded much more intact LP product than cleavage of MBP/LP expressed without zinc. These data indicate a structural role of zinc that allows MBP/LP to fold in a manner such that it is resistant to proteolytic degradation.
A sequence-specific monoclonal antibody directed against the gap junction protein connexin43 (Cx43) is shown here to be specific for the unphosphorylated form of this protein. In tissues and cultured cells containing different phosphorylated and unphosphorylated forms of Cx43, the antibody detected only the latter as shown by Western blotting of native and alkaline phosphatase-treated samples. Immunohistochemically, this monoclonal antibody did not recognize gap junctions in the vast majority of cultured cardiac myocytes, where nearly all detectable Cx43 is phosphorylated. In contrast, it was able to detect some intracellular Cx43 in tracheal smooth muscle cells and an epithelial cell line (Cl-9 cells), producing patterns of labeling consistent with those seen using a polyclonal antibody that recognizes both phosphorylated and unphosphorylated forms of Cx43. Immunostaining of gap junctions in the cultured cells indicates that both phosphorylated and unphosphorylated Cx43 are present in some assembled gap junctions, suggesting that assembled junctions do not contain exclusively the phosphorylated form of the protein. Annular gap junctions, believed to form as part of the pathway for internalization and degradation of gap junctions, were only occasionally and sparsely labeled by the monoclonal antibody, indicating that complete protein dephosphorylation is not required for uptake and degradation of gap junctions. Furthermore, the ability of this antibody to recognize only unphosphorylated Cx43, and not any of the phosphorylated forms present in the tissues and cell types examined, suggests that a unique phosphorylation site, perhaps present in the epitope recognized by this antibody, must be phosphorylated prior to phosphorylation of Cx43 at other sites.
BACKGROUND: Changes in cell shape and motility are important manifestations of oncogenic transformation, but the mechanisms underlying these changes and key effector molecules in the cytoskeleton remain unknown. The Fos oncogene induces expression of ezrin, the founder member of the ezrin/radixin/moesin (ERM) protein family, but not expression of the related ERM proteins, suggesting that ezrin has a distinct role in cell transformation. ERM proteins have been suggested to link the plasma membrane to the actin-based cytoskeleton and are substrates and anchoring sites for a variety of protein kinases. Here, we examined the role of ezrin in cellular transformation. RESULTS: Fos-mediated transformation of Rat-1 fibroblasts resulted in an increased expression and hyperphosphorylation of ezrin, and a concomitant increased association of ezrin with the cortical cytoskeleton. We tagged ezrin with green fluorescent protein and examined its distribution in normal and Fos-transformed fibroblasts: ezrin was concentrated at the leading edge of extending pseudopodia of Fos-transformed Rat-1 cells, and was mainly cytosolic in normal Rat-1 cells. Functional ablation of ezrin by micro-CALI (chromophore-assisted laser inactivation) blocked plasma-membrane ruffling and motility of Fos-transformed fibroblasts. Ablation of ezrin in normal Rat-1 cells caused a marked collapse of the leading edge of the cell. CONCLUSIONS: Ezrin plays an important role in pseudopodial extension in Fos-transformed Rat-1 fibroblasts, and maintains cell shape in normal Rat-1 cells. The increased expression, hyperphosphorylation and subcellular redistribution of ezrin upon fibroblast transformation coupled with its roles in cell shape and motility suggest a critical role for ezrin in oncogenic transformation.
Human recombinant ezrin, or truncated forms, were coated in microtiter plate and their capacity to bind actin determined. F-actin bound ezrin with a Kd of 504 +/- 230 nM and a molecular stoichiometry of 10.6 actin per ezrin. Ezrin bound both alpha- and beta/gamma-actin essentially as F-form. F-actin binding was totally prevented or drastically reduced when residues 534-586 or 13-30 were deleted, respectively. An actin binding activity was detected in amino-terminal constructs (ezrin 1-310 and 1-333) provided the glutathione S-transferase moiety of the fusion protein was removed. Series of carboxyl-terminal truncations confirmed the presence of this actin-binding site which bound both F- and G-actin. The F- and G-actin-binding sites were differently sensitive to various chemical effectors and distinct specific ezrin antibodies. The internal actin-binding site was mapped between residues 281 and 333. The association of ezrin amino-terminal fragment to full-length ezrin blocked F-actin binding to ezrin. It is proposed that, in full-length ezrin, the F-actin-binding site required the juxtaposition of the distal-most amino- and carboxyl-terminal residues of the ezrin molecule.
The effects of dietary administration of diallyl sulfide (DAS), diallyl disulfide (DADS) and allyl mercaptan (AM) on the genotoxicity of different chemicals were studied in two experimental systems: (i) measurement of hepatic DNA single-strand breaks induced in rats by aflatoxin B1 (AFB1), N-nitrosodimethylamine (NDMA) or methylnitrosourea (MNU); (ii) mutagenicity of AFB1 or NDMA on Salmonella typhimurium TA100 using hepatic S9 from rats fed allyl sulfides as the activation system. All compounds strongly reduced hepatic DNA breaks induced by AFB1 and NDMA but did not modify the genotoxicity of MNU. In the Ames test, the mutagenicity of NDMA was strongly inhibited by hepatic S9 from rats fed either compound. The mutagenicity of AFB1 was also reduced but to a lesser extent. Such effects are likely related to the modulation of drug-metabolizing enzymes which play a key role in metabolic activation as well as detoxication of NDMA and AFB1.