RÉSUMÉ
Kawasaki disease (KD) is an acute pediatric vasculitis of unknown etiology that can cause coronary artery aneurysms, and is the leading cause of acquired heart disease in children. We studied aspects of the innate and adaptive immune response in 17 acute KD children prior to treatment with intravenous immunoglobulin. Distinct patterns within the innate immune response correlated with specific clinical features. Proinflammatory myeloid dendritic cells (mDC) were abundant in four of 17 (23·5%) subjects who were older and manifested severe inflammation with clinical myocarditis and elevated hepatobiliary enzyme levels. Of the nine subjects with low levels of anti-inflammatory, tolerogenic mDC, six had enlarged cervical lymph nodes at diagnosis. In contrast, the adaptive immune repertoire varied greatly with no discernible patterns or associations with clinical features. Two subjects with aneurysms had numerous circulating CD8+ T cells. Ten subjects showed low CD4+ T cell numbers and seven subjects had CD4+ T cells in the normal range. CD4+ T cells expressed interleukin-7 receptor (IL-7R), suggesting repeated antigenic stimulation. Thymic-derived regulatory T cells (nTreg ) and peripherally induced regulatory T cells (iTreg ) were also enumerated, with the majority having the nTreg phenotype. Natural killer (NK) and NK T cell numbers were similar across all subjects. Taken together, the results of the immune monitoring suggest that KD may have multiple triggers that stimulate different arms of the innate and adaptive compartment in KD patients. Thus, it is possible that diverse antigens may participate in the pathogenesis of KD.
Sujet(s)
Antigènes/sang , Cellules dendritiques/métabolisme , Lymphocytes/métabolisme , Maladie de Kawasaki/sang , Maladie aigüe , Antigènes/immunologie , Enfant , Enfant d'âge préscolaire , Cellules dendritiques/immunologie , Femelle , Humains , Immunoglobulines par voie veineuse/administration et posologie , Nourrisson , Lymphocytes/immunologie , Mâle , Maladie de Kawasaki/traitement médicamenteuxRÉSUMÉ
Severe combined immunodeficiency (SCID) is a potentially fatal disorder characterized by defective T- and B-lymphocyte function. We describe a 34-week female twin who had developed feeding intolerance, perioral cyanosis, abdominal distension and neutropenia at 1 month of age. Despite several evaluations including an 'inconclusive' newborn screening result for SCID, the presence of profound lymphopenia was unappreciated. Eventually a diagnosis of SCID in association with adenosine deaminase deficiency was made. This case serves to emphasize the importance of newborn screening for SCID in the context of careful evaluation of clinical and laboratory findings that may be overlooked and result in a delay in the diagnosis of a potentially life-threatening condition.
Sujet(s)
Dépistage néonatal , Immunodéficience combinée grave/diagnostic , Adenosine deaminase/déficit , Maladies chez les jumeaux/diagnostic , Femelle , Humains , Nouveau-néRÉSUMÉ
To understand the status of oropharyngeal yeast colonization in human immunodeficiency virus (HIV) -infected outpatients in the era of highly active antiretroviral therapy (HAART), we conducted a prospective, cross-sectional study from October 2009 to January 2010 at a medical centre in southern Taiwan. Fungal cultures of the oropharyngeal swabs were performed on 327 enrolled patients. At enrolment, 258 (79%) patients had been receiving HAART, and 42 (12.8%), 73 (22.3%) and 212 (64.8%) patients had CD4 cell counts ≤200, 201-350, and >350 cells/mm(3) , respectively. Oral yeast colonization was detected in 193 (59%) patients, among whom 157 (81.3%), 25 (13.0%), and 11 (5.7%) were colonized by a single, two and more than two species, respectively. Multivariate analysis showed that receipt of efavirenz-containing regiments and CD4 cell counts >200 cells/mm(3) were associated with lower risks of oral yeast colonization, while intravenous drug users were at a higher risk. Among the 241 isolates recovered, Candida albicans accounted for 69.7%, followed by C. dubliniensis (9.5%), C. glabrata (8.3%), C. tropicalis (3.3%), C. intermedia (2.1%), C. parapsilosis (1.7%), and 11 other species (5.4%). Overall, 230 (95.4%), 236 (97.9%) and 240 (99.6%) isolates were susceptible to fluconazole, voriconazole and amphotericin B, respectively. In conclusion, colonization by C. dubliniensis has emerged in recent years. In addition to a CD4 cell count ≤200 cells/mm(3) , which is a known risk factor for oropharyngeal yeast colonization in HIV-infected patients that was identified in our previous studies, two risk factors, non-receipt of efavirenz-based combinations and intravenous drug use, were first identified in the present study. Fluconazole remained effective in vitro against the yeasts colonizing the oropharynx in this population.
Sujet(s)
Agents antiVIH/usage thérapeutique , Benzoxazines/usage thérapeutique , Candida/isolement et purification , Infections à VIH/complications , Partie orale du pharynx/microbiologie , Inhibiteurs de la transcriptase inverse/usage thérapeutique , Toxicomanie intraveineuse/complications , Infections opportunistes liées au SIDA/traitement médicamenteux , Infections opportunistes liées au SIDA/microbiologie , Adulte , Alcynes , Antifongiques/usage thérapeutique , Numération des lymphocytes CD4 , Candida/classification , Candida/effets des médicaments et des substances chimiques , Candidose buccale/complications , Candidose buccale/traitement médicamenteux , Candidose buccale/microbiologie , Études transversales , Cyclopropanes , Femelle , Fluconazole/usage thérapeutique , Infections à VIH/traitement médicamenteux , Infections à VIH/immunologie , Infections à VIH/virologie , VIH-1 (Virus de l'Immunodéficience Humaine de type 1) , Humains , Mâle , Adulte d'âge moyen , Patients en consultation externe/statistiques et données numériques , Études prospectives , Taïwan , Jeune adulteRÉSUMÉ
BACKGROUND: Primary percutaneous coronary intervention (PCI) in patients with ST-elevation myocardial infarction (STEMI) significantly reduces mortality and morbidity, particularly when door-to-balloon (D2B) time is < 90 min. We sought to minimize preventable delays by instituting an on-site cardiology team-based approach in the emergency department (ED). METHODS: The on-site group comprised 146 consecutive patients with STEMI undergoing primary PCI after implementation of the on-site strategy. This new patient care model was compared with the conventional care administered before instituting the on-site cardiology team-based strategy in ED, which included 90 patients (interim group) receiving primary PCI at a catheterization room in the same building as the ED, and 147 patients (pre-on-site group) undergoing primary PCI at a catheterization room two blocks away from the ED. RESULTS: Median D2B time decreased from 107 min in the pre-on-site group to 72 min in the interim group, and to 47 min in the on-site group, respectively (p < 0.001). The percentage of D2B times < 90 min increased from 34% to 78% and 96%, respectively among the three groups (p < 0.001). Hospitalization costs were significantly reduced in the on-site and interim vs. pre-on-site groups ($5944, $5999, and $6581, respectively; p = 0.008). In-hospital mortality did not differ significantly among the three groups (4.8%, 2.2%, and 6.1%, respectively; p = 0.387). CONCLUSIONS: Institution of an on-site cardiology team-based approach in the ED significantly reduces D2B time in STEMI patients eligible for primary PCI.
Sujet(s)
Angioplastie coronaire par ballonnet/normes , Services des urgences médicales/normes , Infarctus du myocarde/thérapie , Transfert de patient/normes , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Angioplastie coronaire par ballonnet/statistiques et données numériques , Services des urgences médicales/statistiques et données numériques , Femelle , Humains , Mâle , Adulte d'âge moyen , Transfert de patient/statistiques et données numériques , Taïwan , Facteurs temps , Résultat thérapeutiqueRÉSUMÉ
Studies have shown that the 577R allele of α-actinin-3 (ACTN3) is more prevalent in sprint athletes than in the general population or in endurance athletes. We examined the distribution of ACTN3 R577X (rs 1815739) genotypes and alleles in the Taiwanese general population (603) and in elite sprint swimmers who had participated in international/national events (168). Additionally, 50 pre-adolescent (age 11-13 years) male students and 38 adult males who completed 12-weeks of swimming training, were included in the present study. We found that the frequencies of the R allele were significantly higher in female international sprint swimmers (67.6%) than in national sprint swimmers (50.0%) or in the general population (53.7%). The 25-m performance was significantly improved across the genotypes after swimming training among the pre-adolescent males but not among the adult males. In addition, pre-adolescents with the RR genotype had the best performance both before and after training although not statistically significant. In conclusion, the frequencies of ACTN3 577R allele were significantly higher in female international sprint swimmers than among national sprint swimmers or the general population. Furthermore, male pre-adolescents with either the ACTN3 RX or XX genotype showed a greater improvement in 25-meter swimming performance than those with the RR genotype.
Sujet(s)
Actinine/génétique , Athlètes , Performance sportive , Natation , Adolescent , Adulte , Facteurs âges , Allèles , Enfant , Femelle , Fréquence d'allèle , Génotype , Humains , Mâle , Polymorphisme de nucléotide simple , Facteurs sexuels , Taïwan , Jeune adulteRÉSUMÉ
OBJECTIVE: To examine rubella seroepidemiology, and estimate rates of catch-up immunisation and persistence of antibody titers in pregnant women in Taiwan after mass immunisation. DESIGN: A retrospective study. SETTING: Two medical centres and four regional hospitals specialising in obstetric care. SAMPLE: A total of 43,640 prenatal rubella test results for pregnant women from 2001 to 2008. METHODS: Rubella immunoglobulin G (IgG) antibody assay. MAIN OUTCOME MEASURES: Seronegativity, rate of catch-up immunization, and antibody decline. RESULTS: The seronegativity was 10.9% in all pregnant women. Immigrant women had higher seronegativity than indigenous women (OR 2.86; 95% CI 2.65, 3.01). Indigenous women born prior to implementation of the vaccination programmes were more susceptible (20.1%) to rubella infection than were women born thereafter (6.7%). Rates of seropositive conversion were low in both Taiwanese-born and foreign-born women (11.5 and 30.7%, respectively). The rubella antibody titers for vaccinated Taiwanese women in the 1971-1976 and after-1976 birth cohorts declined by 0.6 and 2.3% per year, respectively. CONCLUSIONS: This study demonstrates high seronegativity of older indigenous and immigrant women, a low catch-up immunisation rate, and the persistence of rubella antibodies in Taiwan after mass vaccination. Our study suggests that a single dose of rubella vaccine in teenagers effectively increased rubella seropositivity during their childbearing years. This finding is useful for countries that lack the resources necessary for a two-dose regimen. We recommend free rubella antibody tests to women of childbearing age and free vaccination as required. All postpartum women testing negative for rubella antibodies should be vaccinated before they leave hospital.
Sujet(s)
Anticorps antiviraux/sang , Complications infectieuses de la grossesse/épidémiologie , Vaccin antirubéoleux , Rubéole/épidémiologie , Émigrants et immigrants , Femelle , Humains , Vaccination de masse/statistiques et données numériques , Grossesse , Complications infectieuses de la grossesse/immunologie , Complications infectieuses de la grossesse/prévention et contrôle , Rubéole/immunologie , Rubéole/prévention et contrôle , Syndrome de rubéole congénitale/épidémiologie , Syndrome de rubéole congénitale/immunologie , Syndrome de rubéole congénitale/prévention et contrôle , Études séroépidémiologiques , Taïwan/épidémiologie , Taïwan/ethnologieRÉSUMÉ
Successful management of aneurysms of complex morphology depends primarily on adjunct use of balloons or stents. However, these two methods are technically demanding and have higher complication rates. As an alternative to these two techniques, we have used a catheter-assisted technique with a number of cases. It is simple, versatile, and less demanding technically. This technique should be considered as an alternative strategy in cases of wide-necked aneurysms.
Sujet(s)
Cathétérisme/instrumentation , Cathétérisme/méthodes , Embolisation thérapeutique/instrumentation , Embolisation thérapeutique/méthodes , Anévrysme intracrânien/thérapie , Adulte , Sujet âgé , Angiographie cérébrale , Femelle , Humains , Anévrysme intracrânien/imagerie diagnostique , Mâle , Adulte d'âge moyenRÉSUMÉ
SETTING: Several matrix metalloproteinase (MMP) polymorphisms favouring the development of lung fibrosis after pulmonary tuberculosis (TB) have been described. OBJECTIVE: To investigate the association of MMP-1, MMP-9 and MMP-12 polymorphisms with the development of fibrosis in pulmonary TB. DESIGN: We studied 49 normal subjects and 98 TB patients. We analysed the association between MMP polymorphisms and clinical indices of lung fibrosis by serial chest radiography for 1 year after completion of treatment. RESULTS: The frequency of the MMP-1(-1607G) polymorphism was significantly higher in TB patients with moderate to advanced pulmonary fibrosis than in those with minimal to mild fibrosis. Having at least one -1607G MMP-1 polymorphism increased the risk of moderate and advanced fibrosis respectively by 5.04 (95%CI 1.25-20.30) and 9.87 (95%CI 2.39-40.88) fold. There was no association of MMP-9(-1562T) and MMP-12(Asn357Ser) polymorphisms with lung fibrosis. The production of MMP-1 from monocytes stimulated by interleukin-1 beta was increased in subjects with the 1G allele genotype compared to the 2G/2G genotype. CONCLUSIONS: Patients with MMP-1(-1607G) polymorphism are more vulnerable to more extensive lung fibrosis 1 year after anti-tuberculosis treatment. This may be related to increased MMP-1 activity, leading to enhanced destruction of the matrix with subsequent fibrosis.
Sujet(s)
Prédisposition génétique à une maladie , Poumon/anatomopathologie , Matrix metalloproteinase 1/génétique , Tuberculose pulmonaire/complications , Adulte , Sujet âgé , Antituberculeux/usage thérapeutique , Femelle , Fibrose , Études de suivi , Humains , Mâle , Matrix metalloproteinase 12/génétique , Matrix metalloproteinase 9/génétique , Adulte d'âge moyen , Polymorphisme génétique , Études prospectives , Facteurs de risque , Facteurs temps , Tuberculose pulmonaire/traitement médicamenteuxRÉSUMÉ
This multicenter study evaluated the mutation spectrum and frequencies of the MLH1 and MSH2 genes and determined the occurrence of large genomic deletions in 93 unrelated Taiwanese families that fulfilled the Amsterdam criteria II by denaturing high-performance liquid chromatography analysis, DNA sequencing for aberrant chromatograms, and multiplex ligation-dependent probe amplification analysis. In total, 38 pathogenic mutations (10 large deletions and 28 point mutations or small deletion/insertions) in the MSH2 or MLH1 gene were identified in 61 of the 93 families (66%). Three of the 10 large deletions and 14 of the 28 point mutations or small insertions/deletions have not been reported elsewhere. Three mutations in the MLH1 gene, the MLH1c.1846_1848delAAG (5 families), deletion exons 11-15 (4 unrelated families), and MLH1c.793C>T (13 unrelated families), accounted for 35% of all cases with pathogenic mutations. Haplotype analysis indicated that mutant c.793C>T alleles were derived from two distinct common founders that might be inherited from a single ancestor of presumably Chinese origin. As a mutation detection strategy for Taiwanese Lynch syndrome patients, we recommend that diagnosis starts with screening for large genomic deletions and continues by screening for common mutations in exons 10 and 16 of the MLH1 gene prior to searching for small mutations in the remaining exons.
Sujet(s)
Protéines adaptatrices de la transduction du signal/génétique , Tumeurs colorectales héréditaires sans polypose/génétique , Effet fondateur , Protéine-2 homologue de MutS/génétique , Protéines nucléaires/génétique , Analyse de mutations d'ADN , Santé de la famille , Femelle , Fréquence d'allèle , Mutation germinale , Humains , Mâle , Protéine-1 homologue de MutL , Pedigree , Mutation ponctuelle , Délétion de séquence , TaïwanSujet(s)
Hémoglobines anormales/génétique , alpha-Thalassémie/génétique , Diagnostic différentiel , Sous-unités de l'hémoglobine/génétique , Hétérozygote , Homozygote , Humains , Phénotype , Réaction de polymérisation en chaîne/méthodes , Analyse de séquence d'ADN , Taïwan/épidémiologie , alpha-Thalassémie/sang , alpha-Thalassémie/diagnosticRÉSUMÉ
Inherited factor XIII (FXIII) deficiency is a rare bleeding disorder that can present with umbilical bleeding during the neonatal period, delayed soft tissue bruising, mucosal bleeding and life-threatening intracranial haemorrhage. FXIII deficiency has also been associated with poor wound healing and recurrent miscarriages. FXIII plays an integral role in haemostasis by catalysing the cross-linking of fibrin, platelet membrane and matrix proteins throughout thrombus formation, thus stabilizing the blood clot. The molecular basis of FXIII deficiency is characterized by a high degree of heterogeneity, which contributes to the different clinical manifestations of the disease. There have been more than 60 FXIII mutations identified in the current literature. In addition, single nucleotide polymorphisms have been described, some of which have been shown to affect FXIII activity, contributing further to the heterogeneity in patient presentation and severity of clinical symptoms. Although there is a lifelong risk of bleeding, the prognosis is excellent when current prophylactic treatment is available using cryoprecipitate or plasma-derived FXIII concentrate.
Sujet(s)
Déficit en facteur XIII/génétique , Facteur XIII/physiologie , Hémorragie/sang , Avortement spontané/sang , Avortement spontané/prévention et contrôle , Sujet âgé , Sujet âgé de 80 ans ou plus , Substitution d'acide aminé , Coagulation sanguine/physiologie , Inhibiteurs des facteurs de la coagulation sanguine/sang , Tests de coagulation sanguine , Analyse de mutations d'ADN , Facteur VIII/administration et posologie , Facteur XIII/composition chimique , Facteur XIII/génétique , Facteur XIII/usage thérapeutique , Déficit en facteur XIII/sang , Déficit en facteur XIII/diagnostic , Femelle , Fibrinogène/administration et posologie , Hémorragie/traitement médicamenteux , Humains , Nouveau-né , Mâle , Adulte d'âge moyen , Mutation , Plasma sanguin , Polymorphisme de nucléotide simple/génétique , Grossesse , Complications hématologiques de la grossesse/sang , Complications hématologiques de la grossesse/traitement médicamenteux , Indice de gravité de la maladie , Cicatrisation de plaie/physiologieRÉSUMÉ
Molecular cytogenetic analysis identified a new type of dicentric chromosome involving different breakpoints at 18q in a female fetus. The chromosome anomaly was designated as an asymmetrical pseudoisodicentric chromosome 18, 46,XX,psu dic(18)(pter-->q11.2::q21.3-->pter)mat. A series of BAC clones for 18q11.2 and q21.3 regions were used to identify one breakpoint within the region q11.2 between 19.8 and 21.6 Mb from the telomere of 18p and another breakpoint within q21.3 between 55.4 and 56.9 Mb from the telomere of 18p by FISH analysis. Real-time quantitative PCR and microsatellite analysis further verified that the dicentric chromosome was maternal in origin and resulted from a break-reunion between sister chromatids of a single maternal chromosome. We propose that a loop-type configuration of sister chromatids took place and that the break-reunion occurred at cross sites of the loop to form an asymmetrical isodicentric chromosome during either mitosis or meiosis. In this case, the asymmetrical pseudoisodicentric resulted in an 18pter--> q11.2 duplication and an 18q21.3-->qter deletion, which could have led to certain dysmorphic features of 18q- syndrome in this fetus.
Sujet(s)
Aberrations des chromosomes , Chromosomes humains de la paire 18/génétique , Adulte , Chromosomes artificiels de bactérie , Clones cellulaires , Femelle , Foetus/malformations , Humains , Hybridation fluorescente in situ , Caryotypage , Répétitions microsatellitesRÉSUMÉ
Effective stroke therapies require recanalization of occluded cerebral blood vessels; however, early reperfusion can cause BBB (blood-brain barrier) injury, leading to cerebral oedema and/or devastating brain haemorrhage. These complications of early reperfusion, which result from excess production of ROS (reactive oxygen species), significantly limit the benefits of stroke therapies. Here, we summarize some of the findings that lead to the development of a novel animal model that facilitates identification of specific free radical-associated components of the reperfusion injury process and allows therapeutic interventions to be assessed. In this model, KO (knockout) mice containing 50% activity of the mitochondrial antioxidant manganese-SOD (superoxide dismutase) (SOD2-KO) undergo transient focal ischaemia followed by reperfusion. These animals have delayed (>24 h) BBB breakdown associated with activation of matrix metalloproteinase-9, inflammation and a high brain haemorrhage rate. These adverse consequences are absent from wild-type littermates, SOD2 overexpressors and minocycline-treated SOD2-KO animals. In addition, using microvessel isolations following in vivo ischaemia/reperfusion, we were able to show that the tight junction membrane protein, occludin, is an early and specific target in ROS-mediated microvascular injury. This new model is ideal for studying ischaemia/reperfusion-induced vascular injury and secondary brain damage and offers a unique opportunity to evaluate free radical-based neurovascular protective strategies.
Sujet(s)
Lésions encéphaliques/physiopathologie , Lésion d'ischémie-reperfusion/physiopathologie , Animaux , Anti-inflammatoires/usage thérapeutique , Modèles animaux de maladie humaine , Endothélium vasculaire/anatomopathologie , Humains , Matrix metalloproteinase 9/métabolisme , Souris , Minocycline/usage thérapeutique , Espèces réactives de l'oxygène , Jonctions serrées/anatomopathologieRÉSUMÉ
BACKGROUND AND OBJECTIVES: The demand for transfusions has increased rapidly in southern Taiwan. Between 1993 and 2003, requests for fresh-frozen plasma (FFP) in particular rose dramatically at Kaohsiung Medical University Hospital (KMUH). Transfusion orders were not tightly regulated, and inappropriate use of blood products was common. MATERIALS AND METHODS: We carried out a prospective analysis of transfusion requests from October 2003 to January 2004 at KMUH, and then repeated the audit for another 3-month period after the clinical faculty had undergone five sessions of education on transfusion guidelines. Later, our consultant haematologist applied computerized guidelines to periodic audits. RESULTS: A 5.2% decrease in inappropriate FFP usage followed the educational programme and a further 30% reduction took place after the application of computerized transfusion guidelines. With the guidelines and periodic audits, FFP transfusions decreased by 74.6% and inappropriate requests from 65.2% to 30%. CONCLUSIONS: Hospital policy, computerized transfusion guidelines and periodic audits greatly reduced inappropriate FFP transfusions. An educational campaign had a more limited effect.
Sujet(s)
Transfusion de composants du sang/statistiques et données numériques , Audit médical , Plasma sanguin , Guides de bonnes pratiques cliniques comme sujet/normes , Transfusion de composants du sang/normes , Humains , Études prospectives , Assurance de la qualité des soins de santé , TaïwanRÉSUMÉ
An 8-year-old boy presenting with hypotonia, moderate mental retardation, developmental delay, and psychomotor retardation is reported. Magnetic resonance imaging of the brain at age 3 years revealed a Dandy-Walker variant. Cytogenetic analysis of the peripheral blood revealed a derivative chromosome 12 with unknown additional material attached to the distal region of the long arm of chromosome 12. The parental karyotypes were normal. Spectral karyotyping (SKY) using the 24-color SKY probes and fluorescence in situ hybridization (FISH) using the specific 7p, 7q, 12p, and 12q telomeric probes confirmed a duplication of distal 7p and a deletion of terminal 12q. The karyotype of the proband was designated as 46,XY.ish der(12)t(7;12) (p21.2;q24. 33)(SKY+, 7pTEL+, 12qTEL-). The present case provides evidence for the association of partial trisomy 7p (7p21.2-->pter) and partial monosomy 12q (12q24.33-->qter) with a cerebellar malformation and the usefulness of SKY and FISH in the identification of a de novo aberrant chromosome resulting from an unbalanced translocation.
Sujet(s)
Chromosomes humains de la paire 12/génétique , Chromosomes humains de la paire 7/génétique , Syndrome de Dandy-Walker/génétique , Monosomie , Trisomie , Enfant , Peinture chromosomique , Syndrome de Dandy-Walker/anatomopathologie , Conseil génétique , Humains , Hybridation fluorescente in situ , Caryotypage , Mâle , Translocation génétiqueRÉSUMÉ
OBJECTIVE AND DESIGN: Hydroxamic-and carboxylic-acid based matrix metalloproteinase inhibitors (MMPIs) were compared for their potency against various MMPs, pharmacodynamic properties and in vivo efficacy in a model of cartilage degeneration. MATERIALS AND METHODS: The MMPIs were evaluated for their ability to inhibit human MMPs using the quenched fluorescence assay. The ability of the MMPIs to inhibit the degeneration of the knee joint was evaluated in rats injected intraarticularly with iodoacetate. The amount of MMPI in the plasma and cartilage was determined using liquid chromatography/mass spectrometry/mass spectrometry (LC/ MS/MS). Plasma protein binding was measured by ultrafiltration and unbound MMPI was quantitated using HPLC. RESULTS: The hydroxamic acid based inhibitor PGE-3321996 and the carboxylic acids PGE-2909492 and PGE-6292544 were potent MMP-13 inhibitors, but only the hydroxamic acid PGE 3321996 demonstrated significant inhibition of knee degeneration in the rat iodoacetate model. Both of the carboxylic acids demonstrated superior pharmacokinetic properties and established much higher plasma concentrations than the hydroxamic acid. However, neither of the carboxylic acids was detectable in the cartilage, whereas, the hydroxamic acid was present in both the cartilage and the plasma. The carboxylic acid based MMPIs also demonstrated higher plasma protein binding (>99%) than the hydroxamic acid (79%). CONCLUSIONS: Carboxylic acid-based MMPIs were identified that had superior in vivo plasma exposure compared to a hydroxamic acid inhibitor but lacked in vivo efficacy in the rat iodoacetate model of cartilage degeneration. The lack of in vivo efficacy of the carboxylic acid based MMPIs were probably due to their lack of cartilage penetration which was related to their physicochemical properties.
Sujet(s)
Acides aminés/pharmacocinétique , Acides aminés/usage thérapeutique , Acides carboxyliques , Acides hydroxamiques/pharmacocinétique , Acides hydroxamiques/usage thérapeutique , Inhibiteurs de métalloprotéinases matricielles , Arthrose/traitement médicamenteux , Phénylalanine/analogues et dérivés , Inhibiteurs de protéases , Sulfonamides/pharmacocinétique , Sulfonamides/usage thérapeutique , Sulfones/pharmacocinétique , Sulfones/usage thérapeutique , Acides aminés/composition chimique , Animaux , Acides carboxyliques/composition chimique , Acides carboxyliques/pharmacocinétique , Acides carboxyliques/usage thérapeutique , Cartilage/composition chimique , Cartilage/anatomopathologie , Modèles animaux de maladie humaine , Humains , Acides hydroxamiques/composition chimique , Iodo-acétates/toxicité , Articulation du genou/anatomopathologie , Mâle , Structure moléculaire , Arthrose/induit chimiquement , Arthrose/anatomopathologie , Phénylalanine/composition chimique , Phénylalanine/pharmacocinétique , Phénylalanine/usage thérapeutique , Plasma sanguin/composition chimique , Inhibiteurs de protéases/composition chimique , Inhibiteurs de protéases/pharmacocinétique , Inhibiteurs de protéases/usage thérapeutique , Rats , Rat Sprague-Dawley , Sulfonamides/composition chimique , Sulfones/composition chimiqueRÉSUMÉ
A minilibrary was constructed from DOP-PCR products using microdissected Y-chromosomes of Indian muntjac as DNA templates. Two microclones designated as IM-Y4-52 and IM-Y5-7 were obtained from negative screening of all three cervid satellite DNAs (satellites I, II, and IV). These two microclones were 295 and 382 bp in size, respectively, and shared approximately 70% sequence homology. Southern blot analysis showed that the IM-Y4-52 clone was repetitive in nature with an approximately 0.32-kb register in HaeIII digest. Sequence comparison revealed no similarities to DNA sequences deposited in the GenBank database, suggesting that the microclone sequences were from a novel satellite DNA family designated as cervid satellite V. A subclone of an Indian muntjac BAC clone which screened positive for IM-Y4-52 had a 3,325-bp insert containing six intact monomers, four deleted monomers, and two partial monomers. The consensus sequence of the monomer was 328 bp in length and shared more than 80% sequence homology with every intact monomer. A zoo blot study using IM-Y4-52 as a probe showed that the strong hybridization with EcoRI digested male genomic DNA of Indian muntjac, Formosan muntjac, Chinese muntjac, sambar deer, and Chinese water deer. Female genomic DNA of Indian muntjac, Chinese water deer, and Formosan muntjac also showed positive hybridization patterns. Satellite V was found to specifically localize to the Y heterochromatin region of the muntjacs, sambar deer, and Chinese water deer and to chromosome 3 of Indian muntjac and the X-chromosome of Chinese water deer.
Sujet(s)
ADN satellite , Muntiacus/génétique , Chromosome Y/génétique , Animaux , Séquence nucléotidique , Évolution biologique , Chromosomes/ultrastructure , Femelle , Hybridation fluorescente in situ , Caryotypage , Mâle , Microdissection , Modèles génétiques , Séquences répétées d'acides nucléiques , Similitude de séquences d'acides nucléiquesRÉSUMÉ
BACKGROUND: Calreticulin (CRT), an endoplasmic reticulum protein, has been reported to be essential for the differentiation of neuroblastoma (NB) cells, suggesting that CRT may affect the tumor behavior of neuroblastoma. The aim of this study was to evaluate the association of clinicopathologic factors and patient survival with the expression of CRT in patients with NB. PATIENTS AND METHODS: Sixty-eight NBs were investigated by immunohistochemical staining against CRT, and were divided into positive and negative immunostaining groups. Correlations between calreticulin expression, various clinicopathologic and biologic factors, and patient survival were studied. In seven tumor samples, CRT mRNAs and proteins were evaluated with real-time PCR and western blot, respectively, and correlated with immunohistochemical findings. RESULTS: Among 68 NBs, 32 (47.1%) showed positive CRT expression. Positive CRT immunostaining strongly correlated with differentiated histologies, as well as known favorable prognostic factors such as detected from mass screening, younger age (< or =1 year) at diagnosis and early clinical stages, but inversely correlated with MYCN amplification. Kaplan-Meier analysis revealed that NB patients with CRT expression did have better survival. Multivariate analysis demonstrated CRT expression to be an independent prognostic factor. Moreover, CRT expression also predicted better survival in patients with advanced-stage NBs, and its absence predicted poorer survival in patients whose tumor had no MYCN amplification. The amount of CRT mRNAs and proteins in NB tumor samples tested correlated well with the immunohistochemical expressions. CONCLUSIONS: CRT expression correlates with the differentiation of NB and predicts favorable survival, thereby suggesting CRT to be a useful indicator for planning treatment of NB.
Sujet(s)
Marqueurs biologiques tumoraux/analyse , Calréticuline/biosynthèse , Analyse de profil d'expression de gènes , Neuroblastome/génétique , Neuroblastome/anatomopathologie , Différenciation cellulaire , Enfant , Enfant d'âge préscolaire , Femelle , Études de suivi , Humains , Immunohistochimie , Nourrisson , Nouveau-né , Mâle , Réaction de polymérisation en chaîne , Pronostic , Analyse de survieRÉSUMÉ
The present study was designed to examine whether different p53 haplotypes of exon 4-intron 3-intron 6 affect the frequency of mutations and loss of heterozygosity (LOH) of the p53 gene in male oral squamous cell carcinomas (OSCCs) in Taiwan. We found that individuals without two Pro-W-G alleles had significantly higher frequency of p53 mutations than those with two Pro-W-G alleles (odds ratio (OR) = 1.98; 95% confidence interval (CI), 1.10-3.56). Out of the 172 p53 gene exon 4 informative male OSCCs, 72 (41.9%) showed LOH. Among these 72 OSCCs with LOH, the frequency of Pro allele loss was 73.6% (53/72). It is notable that alcohol drinking increased the frequency of Arg allele loss (OR = 10.56; 95% CI, 1.23-234.94) in OSCCs from patients who both smoked cigarettes and chewed areca quid (AQ). The frequency of LOH of p53 was not different between p53-mutated OSCCs and p53-normal OSCCs. Thus, the present study revealed that (a) the Arg allele is associated with p53 mutations, (b) the Pro allele is preferentially lost in OSCCs associated with cigarette smoking and AQ chewing, while the frequency of Arg allele loss is increased with alcohol drinking, and (c) haploinsufficiency of p53 is in itself likely to contribute to tumour progression in Taiwanese OSCCs.