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Biomedical subjects

S Martin

Publications and source records attributed to S Martin.

At least 145 records · Page 8Linked to original sources

Isolation of genomic DNA from blood using a novel filter-based DNA purification technology.

A novel DNA purification technology is described that enables the purification of pure dsDNA from blood. When compared to existing DNA purification methods, the Whatman BioScience Purification System (WBPS) offers a fast and convenient way to recover high yields of DNA. WBPS is based on a unique filter system that entraps DNA within a matrix. This allows the process to be performed in a single unidirectional reaction vessel, reducing user interaction and multiple centrifugation steps.

DNA↗

[Recurrent syncope and Chiari malformation].

Downward herniation of the cerebellar tonsils through the foramen magnum (Chiari I malformation) is usually revealed by head and neck pain, often associated with brain-stem or spinal cord disturbances. Syncopes are rarely reported and may be difficult to link to their cause when they occur alone. We report two cases with brief and repetitive syncopes revealing a Chiari I malformation. These manifestations may be attributed to transient compression of brain stem or vascular structures at the craniocervical junction, triggered by intracranial pressure increase, as they disappear after posterior fossa decompression.

Budd-Chiari Syndrome↗

A randomised controlled trial of delayed antibiotic prescribing as a strategy for managing uncomplicated respiratory tract infection in primary care.

BACKGROUND: Despite evidence that uncomplicated lower respiratory tract infection (cough) does not respond appreciably to antibiotics and that bacterial resistance is increasing, general practitioners (GPs) still prescribe frequently. AIM: To assess delayed antibiotic prescribing as a strategy for reducing the unnecessary use of antibiotics for cough in primary care. DESIGN OF STUDY: Open randomised controlled trial of delayed versus immediate prescribing of antibiotics. SETTING: One hundred and ninety-one adult patients with uncomplicated cough in 22 Scottish practices who would have received antibiotics under the GP's usual practice were randomised to receive either an immediate prescription (92 patients) or a delayed prescription (99 patients). METHOD: Delayed subjects were asked to wait a week before deciding whether to collect their prescription. Outcome measures included symptom duration, prescription uptake, patient satisfaction, patient enablement, and subsequent consultation rates. The 48 GPs who recruited patients were surveyed six months after the trial to see whether they used delayed prescribing as a part of their normal practice. RESULTS: Study and control groups were similar at baseline. Of the subjects in the delayed arm, 55% did not pick up their prescription. Although most patients were satisfied, more patients in the immediate arm were very satisfied with the treatment (P = 0.001) and the consultation (P = 0.03). The patients in the immediate arm were also more enabled (3.3 versus 2.4; P = 0.04), although more of them intended to consult for similar complaints in the future (85% versus 69%, P = 0.02). We were unable to detect any difference in actual consulting behaviour in the follow-up period (mean = 15 months [SD = 5 months]). Subsequently, 68% of GPs used delayed prescribing at least monthly; all gave the prescription to the patient. CONCLUSION: Delayed prescribing is effective at reducing the use of antibiotics for self-limiting cough; however, patients are less satisfied and enabled as a result. Patients may be deterred from consulting rather than becoming enabled.

Adult↗

The association between perineal trauma and spontaneous perineal tears.

OBJECTIVE: We assessed whether women who had a perineal trauma (episiotomy or spontaneous tear of the second degree or higher) at the first delivery were at increased risk for spontaneous perineal tears at the next delivery, and whether the risk increases with the severity of previous perineal trauma. DESIGN: Retrospective cohort study. POPULATION: We included data from 1895 women who had their first and second deliveries at Saint-Sacrement Hospital, Quebec City, Canada, between 1985 and 1994. Our study was restricted to women who gave birth vaginally to a single living neonate at their first 2 deliveries and who did not have an episiotomy at the second delivery. We extracted the data from the Department of Obstetrics computerized database. OUTCOMES MEASURED: Spontaneous perineal tears (of second degree or higher) at the second delivery. RESULTS: Having a perineal trauma at the first delivery more than tripled the risk (relative risk=3.3; 95% confidence interval, 2.6-4.2) of spontaneous perineal tears at the second delivery. The risk of spontaneous perineal tears at the second delivery increased with the severity of previous perineal trauma at birth. CONCLUSIONS: Our results show that the risk of spontaneous perineal tears at subsequent deliveries increases with the presence and the severity of perineal trauma at the first delivery.

Adult↗

UVB-irradiated dendritic cells are impaired in their APC function and tolerize primed Th1 cells but not naive CD4+ T cells.

We have shown that low-dose UVB radiation converts Langerhans cells (LC) from immunogenic to tolerogenic APC. Therefore, we questioned whether low-dose UVB irradiation of bone marrow-derived dendritic cells (DC) alters their APC function, thereby inducing tolerance in T cells. To address this issue, cocultures of DC; and naive, allogeneic T cells; naïve, OVA-specific TCR-transgenic T cells from DO11.10 mice; or primed, antigen-specific T cells using the Th1 clone AE7 were analyzed. First, we found low-dose UVB-irradiated DC (UVB-DC) to dose-dependently (50-200 J/m2) inhibit T-cell proliferation of naive and primed T cells. In addition, supernatants harvested from cocultures of UVB-DC and naive T cells showed markedly reduced levels of IL-2 and IFN-gamma and to a lesser degree of IL-4 and IL-10, suggesting a preferential down-regulation of Th1 responses by UVB-DC. FACS analysis of UVB-DC revealed no changes in surface expression of MHC, costimulatory, and adhesion molecules. To test tolerance induction, allo- or antigen-specific T cells isolated from cocultures with unirradiated DC and UVB-DC were restimulated with unirradiated DC or IL-2. It is interesting that UVB-DC induced antigen-specific tolerance in the Th1 clone AE7. In contrast, UVB-DC induced a partial inhibition of allogeneic T-cell proliferation but no tolerance with similar unresponsiveness to restimulation with IL-2 and unirradiated DC irrespective of their haplotype. Similar observations were made when naive, TCR-transgenic T cells from DO11.10 mice were used. In conclusion, UVB-DC are impaired in their APC function and tolerize the primed antigen-specific Th1 clone AE7 but not naive allo- or OVA-specific T cells.

Animals↗

Correlation of collagen organization with polarization sensitive imaging of in vitro cartilage: implications for osteoarthritis.

OBJECTIVE: Optical coherence tomography (OCT), a new method of high resolution imaging, has shown feasibility for assessing articular cartilage to identify early changes in osteoarthritis (OA) and monitor therapy. OCT is analogous to ultrasound, measuring the intensity of backreflected infrared light rather than sound. The resolution of this technology is up to 25 times higher than existing methods. We investigated the correlation between changes observed by OCT and the degree of collagen organization in OA cartilage. METHODS: Polarization sensitive OCT (PSOCT) imaging was used to assess changes in cartilage collagen organization in vitro. RESULTS: The presence (or absence) of PSOCT changes correlated with collagen organization (or disorganization) on histology as assessed by picrosirius polarization microscopy (no significant difference). In multiple cases, cartilage was abnormal by both PSOCT and polarization microscopy, but was grossly normal by routine staining, showing cartilage thickness > 2 mm and no fibrillations. CONCLUSION: This in vitro study suggests PSOCT changes in cartilage are due to the state of collagen organization. The combination of high resolution structural imaging and birefringence detection make OCT a potentially powerful technology for early assessment of OA.

Cartilage, Articular↗

Total body 4.5 Gy gamma irradiation-induced early delayed learning and memory dysfunction in the rat.

In an attempt to determine the consequences of total body radiation damage on learning and memory in the rat, twenty-eight male Wistar rats aged 4 months received 4.5 Gy total body gamma-irradiation (TBI) while 28 rats received sham irradiation. Sequential behavioral studies of negative reinforcement including a/ one- and b/ two-way avoidance tasks were undertaken. a/ One-way avoidance test: this test was performed before and after TBI. Prior to irradiation both groups were similar. At 20 days (D) and at 3 months post-TBI, irradiated rats had a significantly lower percentage of avoidance than controls but no statistical difference was found at 5 months post-TBI. b/ Two-way avoidance test: this test was performed only after TBI. At days 21, 22, 23, 24, (leaming) and at 4 or 6 months (recalls) post-TBI the mean percentage of avoidance was significantly lower in irradiated than in control rats. This study demonstrates that total-body exposure to 4.5 Gy gamma-irradiation induces behavioral dysfunction affecting learning and transitorily memory. These results suggest that a relatively low dose of total body irradiation can induce neurological complications, which persist 4-6 months later.

Animals↗

Low dose of the gamma acute radiation syndrome (1.5 Gy) does not significantly alter either cognitive behavior or dopaminergic and serotoninergic metabolism.

The aim of this study was to evaluate the early-delayed effects of a low dose of the gamma acute radiation syndrome (1.5 Gy) on memory and on dopaminergic and serotoninergic metabolism in Swiss albino CD1 mice, of various ages (6, 10 and 20 weeks). At different times after irradiation (from 24 hr to three months), the mice were trained in a single-trial passive avoidance task and tested for retention either 24 hr or 5 days later. Their performance was compared to that of mice that were sham-irradiated. At the end of the behavioral test (days 3, 9, 30 and 93), the concentrations of dopamine (DA) and serotonin (5HT) and their metabolites were determined in hippocampus, anterior cortex and striatum of mice irradiated at the age of six weeks. No significant behavioral effect was observed whichever the age of the animals or the delay of observation. On the contrary at the moderate dose of 4.5 Gy we observed a significant memory deficit 9 days after the exposure. Considering the neurochemical study, in the striatum or in the frontal cortex, no significant modification was observed whichever the delay or the molecule. In the hippocampus slight modifications were noted: an increase (+144%, p = 0.002) in DA level on day 3 after exposure, and a decrease (-27%, p = 0.028) of 5HT level on day 30 post-irradiation. These modifications were only transient and not associated to modifications of the catabolites. This study demonstrates that total-body exposure to gamma radiation at low dose seems to induce only slight effects on the central nervous system.

Acute Disease↗

Relation between phosphatidylserine exposure and store-operated Ca(2+) entry in stimulated cells.

A significant increase in intracellular Ca(2+) is required to trigger the remodeling of the cell plasma membrane. Scott syndrome is an extremely rare inherited disorder of the transmembrane migration of phosphatidylserine toward the exoplasmic leaflet in blood cells. We have recently reported a reduced capacitative Ca(2+) entry in Scott cells [Martínez et al. (1999) Biochemistry 38, 10092-10098]. We have investigated here the links between defective phosphatidylserine exposure and Ca(2+) signaling in Scott cells by focusing on the Ca(2+) entry following the emptying of intracellular stores. After depletion of caffeine- or thapsigargin-sensitive stores, Ca(2+) entry was lower in Scott compared to control lymphoblasts. However, the simultaneous depletion of both types of stores restored a normal Ca(2+) influx across the plasma membrane in Scott cells and phosphatidylserine externalization ability was improved concomitantly with capacitative Ca(2+) entry. These observations point to the essential role of capacitative Ca(2+) entry in the control of phosphatidylserine exposure of stimulated cells.

Aged↗

Reduced antibody response to revaccination with meningococcal serogroup A polysaccharide vaccine in adults.

Widespread use of meningococcal A and C polysaccharide (MACP) vaccines has raised concerns about induction of hyporesponsiveness to these polysaccharides. Immunological hyporesponsiveness to C polysaccharide has been clearly documented in infants, children and adults but only limited data from Gambian children are available for A polysaccharide. We investigated whether a second dose of MACP, given 6 months after an initial dose affected the immunological response as measured by the serum bactericidal assay (SBA) and enzyme-linked immunosorbent assay (ELISA), to serogroup A meningococci in young adults (university students, n=36). Serogroup A SBA responses 1 month following the second dose of MACP (geometric mean titre (GMT) 103.6, 95% CI 45.6-235.1) were approximately one third of the levels observed 1 month post first dose (GMT 281.9, 95% CI 134.9-581.4). The serogroup A-specific IgG levels post second dose (GMC 21.2, 95% CI 15.3-29.4) were also significantly lower at an average of three-quarters the level post first dose (GMC 28.7, 95% CI 20.8-39.7). This confirms that revaccination with MACP vaccine, 6 months following the initial dose, results in a reduced immunological response to A polysaccharide in adults. Repeated vaccination with MACP vaccine may be ineffective and development and use of meningococcal serogroup A conjugate vaccines should be encouraged.

Adolescent↗

Fas ligand costimulates the in vivo proliferation of CD8+ T cells.

Fas ligand (FasL/CD95L/APO-1L) is one of a growing number of TNF family members whose triggering costimulates maximal proliferation of activated T cells. In this study we show that maximal Ag-dependent accumulation of transferred TCR-transgenic CD8(+) T cells requires Fas (CD95/APO-1) expression by the adoptive hosts. Additionally, adoptively transferred FasL(+) CD8(+) T cells demonstrate a 2-fold advantage in Ag-driven expansion over their FasL(-)counterparts. This study illustrates the in vivo role of TCR-dependent FasL costimulation in the Ag-specific proliferation of both heterogeneous and homogeneous populations of primary CD8(+) T cells and long-term CTL lines. Thus, cross-linking FasL on naive and Ag-experienced CD8(+) T cells whose Ag-specific TCRs are engaged is required to drive maximal cellular proliferation in vivo.

Adoptive Transfer↗

Ability of 3 different meningococcal C conjugate vaccines to induce immunologic memory after a single dose in UK toddlers.

To test for immunologic memory after a single dose of meningococcal C conjugate (MCC) vaccine in toddlers, 226 children 12-18 months old were randomized to receive 1 of 3 MCC vaccines, with a C polysaccharide booster 6 months later. The protein conjugate was diphtheria mutant toxoid in 2 vaccines (MCC-CRM(197)) and was tetanus toxoid in the third (MCC-TT). One month after the MCC vaccines, 91%-100% of children had serum bactericidal antibody (SBA) titers > or =8, and 89%-100% had a > or =4-fold increase. Geometric mean titer (GMT) increased from <4 to 215 (95% confidence interval [CI], 166-279). MCC-TT induced higher SBA GMTs (P<.001) and higher proportions with SBA > or =8 (P=.02) than did the MCC-CRM(197) vaccines. By 6 months, GMTs had decreased to 55.1 (95% CI, 40-76), but IgG antibody avidity increased (P<.001). Induction of immunologic memory was confirmed by a GMT of 1977 (range, 1535-2547) after the polysaccharide booster and a further increase in avidity. This evidence justified the use of a single dose in a catch-up immunization program for children 1-18 years old.

Antibodies, Bacterial↗

A conserved mechanism of retrovirus restriction in mammals.

The murine Fv1 gene restricts infection by N- or B-tropic murine leukemia viruses at a postentry, preintegration stage. The Fv1-sensitive viruses previously used for the study of Fv1 encode an ecotropic envelope gene and thus only infect rodent cells. Consequently, the study of Fv1 restriction has been carried out solely in mice and murine cell lines. By infection with retroviral vectors containing N- or B-tropic core and pantropic vesicular stomatitis virus-G envelope protein, we now demonstrate that cell lines derived from various mammalian species, including humans, have an Fv1-like retrovirus restriction function, preventing N-tropic vector infection. Like Fv1, restriction is directed at amino acid 110 of the viral capsid protein. In contrast to Fv1, the novel restriction is characterized by the absence of reverse-transcribed viral DNA. We speculate that these activities have been selected for by retroviral epidemics in the distant past.

3T3 Cells↗

Prevalence of vacuolar lesions consistent with scrapie in the brains of healthy cull sheep of the Shetland Islands.

To determine the levels of background scrapie-like pathology in the brains of clinically normal adult sheep, the brains of 1106 sheep from 28 known scrapie-infected flocks and nine apparently uninfected flocks were examined during 1998 and 1999. One per cent of the brains had vacuolar pathology and disease-specific accumulations of prion protein consistent with a diagnosis of scrapie. All the positive animals had at least one allele of the prion protein gene encoding valine at codon 136, and originated from flocks in which cases of clinical scrapie had been confirmed within the last four years. The parasympathetic nucleus of the vagal nerve was the most consistently and severely affected nucleus in the medulla oblongata, suggesting that the infection enters the brain via ascending fibres of the vagus nerve.

Animals↗