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1.
Genet Med ; 23(11): 2096-2104, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34230640

RESUMEN

PURPOSE: Where multiple in silico tools are concordant, the American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) framework affords supporting evidence toward pathogenicity or benignity, equivalent to a likelihood ratio of ~2. However, limited availability of "clinical truth sets" and prior use in tool training limits their utility for evaluation of tool performance. METHODS: We created a truth set of 9,436 missense variants classified as deleterious or tolerated in clinically validated high-throughput functional assays for BRCA1, BRCA2, MSH2, PTEN, and TP53 to evaluate predictive performance for 44 recommended/commonly used in silico tools. RESULTS: Over two-thirds of the tool-threshold combinations examined had specificity of <50%, thus substantially overcalling deleteriousness. REVEL scores of 0.8-1.0 had a Positive Likelihood Ratio (PLR) of 6.74 (5.24-8.82) compared to scores <0.7 and scores of 0-0.4 had a Negative Likelihood Ratio (NLR) of 34.3 (31.5-37.3) compared to scores of >0.7. For Meta-SNP, the equivalent PLR = 42.9 (14.4-406) and NLR = 19.4 (15.6-24.9). CONCLUSION: Against these clinically validated "functional truth sets," there was wide variation in the predictive performance of commonly used in silico tools. Overall, REVEL and Meta-SNP had best balanced accuracy and might potentially be used at stronger evidence weighting than current ACMG/AMP prescription, in particular for predictions of benignity.


Asunto(s)
Genómica , Neoplasias , Simulación por Computador , Variación Genética , Humanos , Mutación Missense , Neoplasias/diagnóstico , Neoplasias/genética
3.
Eur Arch Paediatr Dent ; 16(4): 365-70, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25800500

RESUMEN

AIM: To determine those organisms of the genus Candida associated with dental caries by investigating samples from active carious lesions. Within the genus Candida, the species Candida albicans and Candida dubliniensis are capable of forming chlamydospores and germ tubes. Until it became possible in 1995 to differentiate between the two species taxonomically, C. dubliniensis was falsely identified as C. albicans. Whilst the importance of C. albicans for rapidly progressing early childhood caries (ECC) has been recognised, so far there have been only reports about C. dubliniensis in connection with children/mothers who have been infected with HIV or already developed AIDS. In the present study, C. dubliniensis was for the first time isolated from plaque and carious dentine of a healthy five-year-old boy. METHODS: As part of the investigation, a number of samples were collected from individual children affected by active dental caries. Amongst the samples, one in particular indicated that Candida species might be involved. The patient was a five-year-old boy with ECC of the primary dentition, scheduled for restorative treatment under general anaesthesia. Before treatment, a salivary, plaque (region of 54/55) and soft carious dentine sample from the tooth 51 was taken before extraction. The counts of yeasts, lactobacilli (LB) and mutans streptococci were determined in the samples. RESULTS: The boy's dmft was 11, which was dominated by the d component. In the saliva of the boy, LB and mutans streptococci (MS) were detected. In plaque and carious dentine, MS and most interestingly C. dubliniensis were present. The yeasts were visualised in carious dentine by means of scanning electron micrographs. CONCLUSIONS: Plaque and carious dentine may be a further habitat of C. dubliniensis.


Asunto(s)
Candida/clasificación , Caries Dental/microbiología , Placa Dental/microbiología , Dentina/microbiología , Diente Primario/microbiología , Carga Bacteriana , Candida/aislamiento & purificación , Preescolar , Recuento de Colonia Microbiana , Índice CPO , Humanos , Lactobacillus/clasificación , Lactobacillus/aislamiento & purificación , Lacticaseibacillus casei/aislamiento & purificación , Masculino , Microscopía Electrónica de Rastreo , Saliva/microbiología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Streptococcus mutans/aislamiento & purificación , Streptococcus sobrinus/aislamiento & purificación
4.
J Periodontal Res ; 49(3): 371-81, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23815471

RESUMEN

BACKGROUND AND OBJECTIVE: Recently, porcine acellular dermal matrix (PADM) has been proposed as a possible alternative to autogenous grafts in periodontal plastic surgery. The aim of the present study was to investigate the in vitro responses of four different oral cell lines cultured on a novel PADM. Furthermore, tissue reaction to PADM was evaluated histologically after subcutaneous implantation in mice. MATERIAL AND METHODS: Human gingival fibroblasts (HGF), human osteoblast-like cells, human umbilical vein endothelial cells and human oral keratinocytes (HOK) were cultured and transferred on to the PADM. A tissue culture polystyrene surface served as the control. The viability of all tested cell lines on PADM was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assay and PrestoBlue(®) reagent. The ToxiLight(®) assay was performed to analyze the effect of PADM on adenylate kinase release. PADM was implanted into nude mice subcutaneously and subjected to histological analysis after 21 d. RESULTS: Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assays, all tested cell lines cultured on PADM demonstrated a significant increase of viability compared to the control group (each p < 0.001) with the exception of HGF and HOK after 3 d (each p > 0.05). According to the PrestoBlue(®) analysis, all cell lines demonstrated a significant increase of viability compared to the control group at the particular points of measurement after 18 h (HGF p < 0.01; human osteoblast-like cells, human umbilical vein endothelial cells, HOK each p < 0.001). No significant cytotoxic effects of PADM on the tested cell lines could be observed, as assessed by changes in adenylate kinase release. Subcutaneous implantation of PADM into nude mice demonstrated good integration with surrounding tissues and significant revascularization of its collagen structure. CONCLUSION: Overall, the results suggest that PADM is a promising substitute for autogenous soft tissue grafts in periodontal surgery.


Asunto(s)
Dermis Acelular , Encía/citología , Gingivoplastia/métodos , Andamios del Tejido , Adenilato Quinasa/análisis , Animales , Técnicas de Cultivo de Célula , Línea Celular , Supervivencia Celular/fisiología , Trasplante de Células/métodos , Colorimetría/métodos , Colorantes , Femenino , Fibroblastos/trasplante , Regeneración Tisular Guiada Periodontal/métodos , Células Endoteliales de la Vena Umbilical Humana/trasplante , Humanos , Indicadores y Reactivos , Queratinocitos/trasplante , Ratones , Ratones Desnudos , Osteoblastos/trasplante , Tejido Subcutáneo/cirugía , Porcinos , Sales de Tetrazolio , Tiazoles , Factores de Tiempo , Ingeniería de Tejidos/métodos
5.
Eur J Med Res ; 16(11): 514-8, 2011 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-22027646

RESUMEN

OBJECTIVE: The aim of the present clinical pilot study was to examine the influence of a combination of micronutrients on individuals with high stress experience. METHODS: 40 healthy students (28 female, 12 male) with a mean age of 27.1 ± 3.0 years, experiencing high examination stress, were chosen. After approval of the ethics commission, one group of students (n = 19) took a combination of micronutrients (Orthomol vital m/f) for three months, whereas other students (n = 21) served as control group. All participants underwent at the beginning and at the end of the trial a dental examination, a determination of 10 periodontal pathogens, a salivary and a blood analysis. In addition, the participants filled in a questionnaire on nutrition, quality of life and degree of stress experienced during their final examinations. RESULTS: The evaluation of the results, obtained at the end of the trial period, showed that for all students a slight worsening of oral hygiene and an increased consumption of unhealthy food could be observed. The intake of the micronutrients led to a slight improvement of the degree of gingival inflammation in comparison to the control group. The blood analysis showed an increase in vitamin (vitamin C, vitamin E) and zinc concentrations, and a lower increase in CRP. In the male subjects, a decrease in the serum concentrations of triglycerides (p = 0.073) and LDL (p = 0.048) was observed. CONCLUSIONS: This pilot study shows that micronutrients, taken during periods of high stress experience, had a beneficial effect on inflammatory processes and helped reduce the level of some of the plasma lipids in males, and thus can be recommended for supplementing the diet. However, additional studies with a higher number of subjects, also suffering from periodontal disease, are necessary to show the effect of a micronutrient supplementation more clearly.


Asunto(s)
Micronutrientes/farmacología , Salud Bucal , Adulto , Análisis Químico de la Sangre , Femenino , Encía/efectos de los fármacos , Encía/patología , Humanos , Inflamación/patología , Masculino , Micronutrientes/administración & dosificación , Adulto Joven
6.
Eur Arch Paediatr Dent ; 11(4): 181-6, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20840828

RESUMEN

AIM: This was to determine the prevalence of Lactobacilli (LB) species in different stages of caries progression and are considered as secondary invaders of existing carious lesions and specialists for caries progression. METHODS: Carious dentine samples were collected from 70 primary molars (M) during step-wise (S1, S2: n = 35 M) or one-step (O1: n = 35 M) caries treatment and after 11 months of temporary restorations (S3, O2). LB were identified by selected physiological and biochemical characteristics, ratio of lactic acid isomers, electrophoretic mobilities of lactic acid dehydrogenases, and shotgun mass mapping by MALDI mass spectrometry. RESULTS: LB were isolated from 46% of soft dentine samples (S1). The prevalence of LB from hard dentine collected during caries excavation (O1) reached 34%, after 8 weeks of temporary filling (S2) 11%, and 9% each after 11 months of temporary restoration (S3, O2). The mean total bacterial counts (cfu) of soft dentine (S1) were 3.6 x 105. From hard dentine during caries excavation (O1) 4.4x104 cfu were calculated, at S2 3.7 x 10³ cfu, at S3 0.1 x 10³ cfu, and at O2 1.8 x 10³ cfu. The percentages of LB in the cfu for LB positive dentine samples were for S1 / S2 / S3 / O1 / O2: 60% (16 M)/34% (4 M)/54% (3 M)/57% (9 M), and 64% (3 M). Five LB species were identified from carious dentine: L. paracasei subsp. paracasei, L. paracasei subsp. tolerans, L. rhamnosus, L. gasseri, and L. alimentarius. CONCLUSIONS: While L. rhamnosus and L. paracasei subsp. paracasei occurred in all caries progression stages, the other species were found only sporadically. L. paracasei subsp. paracasei and L. rhamnosus might be the specialists of the LB in carious progression.


Asunto(s)
Caries Dental/microbiología , Lactobacillus/clasificación , Diente Molar/microbiología , Diente Primario/microbiología , Carga Bacteriana , Hidróxido de Calcio/uso terapéutico , Niño , Preparación de la Cavidad Dental/métodos , Recubrimiento de la Pulpa Dental/métodos , Restauración Dental Provisional/métodos , Dentina/microbiología , Progresión de la Enfermedad , Electroforesis en Gel de Poliacrilamida , Estudios de Seguimiento , Humanos , Isomerismo , L-Lactato Deshidrogenasa/análisis , Ácido Láctico/análisis , Lactobacillus/aislamiento & purificación , Lacticaseibacillus rhamnosus/aislamiento & purificación , Materiales de Recubrimiento Pulpar y Pulpectomía/uso terapéutico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
7.
Eur J Med Res ; 13(7): 349-54, 2008 Jul 28.
Artículo en Inglés | MEDLINE | ID: mdl-18700193

RESUMEN

Dental erosion caused by acidic beverages is common and occurs with increasing tendency. The aim of this in vitro study was to analyse the erosive potential of apple juice on human enamel samples from the first and second dentition. Apple-juice-containing beverages (n = 23) were selected, and pH and buffering capacity were determined. Enamel samples were prepared from impacted, surgically removed wisdom teeth (20 mm superset2) and from deciduous teeth (16 mm superset2). Prepared enamel slices were incubated with a selected apple juice (pH = 3.5) for up to 24 h; the amounts of released calcium were determined colorimetrically, and mean surface roughness (Ra) of the enamel was measured using an optical profilometric device (perthometer, Mahr, Göttingen, Germany). Controls were incubated with a 0.9 % sodium chloride solution under the same conditions (37 degrees C, humidified atmosphere of 5% CO subset2 and 95 % air). The surfaces of the enamel samples were visually examined by CLSM (Leica TCS SP2). The pH-values of the apple juices ranged from 3.3 to 4.2. Incubating the enamel slices (from both dentitions) with a selected apple juice caused a time dependent release of calcium. After 24 h, the primary dentition showed Ca-release values of 0.61 +/- 0.035 mg/ 20 mm superset2 and the second dentition of 0.41 +/- 0.085 mg/ 20 mm superset2; the surface roughness for the primary teeth was 6.8 +/- 1.09 microm and for the second dentition 6.2 +/- 0.41 microm. CLSM show structural changes on all surfaces when compared to the controls. In this in vitro study, the erosive potential of apple juice on teeth of the first and second dentition could be demonstrated. However, it must be considered that numerous modifying factors influence the human enamel surface in vivo; therefore, a direct translation from in-vitro conditions can only be done with caution.


Asunto(s)
Esmalte Dental/efectos de los fármacos , Dentición , Adulto , Bebidas , Tampones (Química) , Niño , Preescolar , Esmalte Dental/química , Solubilidad del Esmalte Dental , Humanos , Concentración de Iones de Hidrógeno , Malus , Reproducibilidad de los Resultados , Erosión de los Dientes/etiología , Diente Primario/química
8.
Eur J Med Res ; 12(3): 120-8, 2007 Mar 26.
Artículo en Inglés | MEDLINE | ID: mdl-17507308

RESUMEN

OBJECTIVE: The aim of the present study was to examine antibiotic resistant strains among the implant-associated microorganisms in vitro, first as mixed cultures and again as pure isolates for resistance to one of five antibiotics. METHODS: Samples were taken with sterile paper points from the deepest pocket of one implant per patient (n = 24) to culture the total oral micro-flora. The samples were streaked on agar (Schaedler or BHI) and incubated for 7 d in an anaerobic atmosphere. All colonies were rinsed off the plates, aliquots were added to top-agar. Susceptibility against antibiotics (ampicillin, ampicillin + sulbactam, azithromycin and penicillin, moxifloxacin) was determined using the Etest. Resistant strains were picked, purified and characterized, and the Etests were repeated with a selection of the pure isolates. RESULT: The majority of the mixed cultures (67 - 100 %) showed complete antibiotic resistance. No association with clinical parameters like pocket depth, bleeding on probing or insertion of implants into transplanted bone could be found. Smoking and the surface of the implant also had no influence. 23 % of the 597 resistant colonies contained only yeasts, mostly isolated from irradiated tumour patients. Of the 458 resistant bacteria, the majority were Gram-positive cocci or rods. Staphylococci and M. micros were detected occasionally. The resistance for the 138 selected pure isolates was in most cases lower than for the total micro-flora, irrespective of the antibiotic. CONCLUSIONS: The higher resistance of the total flora might be explained by synergistic interactions between its members.


Asunto(s)
Antibacterianos/farmacología , Bacterias Anaerobias/efectos de los fármacos , Bacterias Anaerobias/aislamiento & purificación , Implantes Dentales/microbiología , Placa Dental/microbiología , Farmacorresistencia Bacteriana , Ampicilina/farmacología , Compuestos Aza/farmacología , Azitromicina/farmacología , Placa Dental/tratamiento farmacológico , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Femenino , Fluoroquinolonas , Bacterias Gramnegativas/aislamiento & purificación , Cocos Grampositivos/aislamiento & purificación , Bacilos Grampositivos/aislamiento & purificación , Humanos , Técnicas In Vitro , Masculino , Pruebas de Sensibilidad Microbiana , Moxifloxacino , Penicilina G/farmacología , Quinolinas/farmacología , Sulbactam/farmacología
9.
Eur J Med Res ; 10(8): 333-8, 2005 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-16131474

RESUMEN

PURPOSE: The aim of this study was to identify microorganisms which might be present in closed periapical lesions and to determine their relationship to conventional clinical parameters. - METHODS: In 11 patients, samples were taken with paper points from 14 teeth with periapical radiolucency, in seven cases two samples were taken from two different sites of the same tooth (n=21). Clinical parameters were determined. The 16S rDNA of eleven bacterial species could be simultaneously detected via a modified polymerase chain reaction (PCR) based technique. - RESULTS: All samples contained more than one bacterial species. Most frequently, M. micros was detected alone or in combination with E. faecalis, P. aeruginosa, E. coli, F. nucleatum or S. sanguinis. When apical palpation was positive, often M. micros and F. nucleatum were both present. Other clinical symptoms like tenderness to percussion were not associated with a particular microflora. - CONCLUSIONS: In closed periapical lesions detected by radiography, specific bacteria were identified by means of a modified PCR technique. No clear associations between clinical symptoms and these bacteria were found. Surgical intervention might be indicated for some of the persistent lesions.


Asunto(s)
Bacterias Anaerobias/aislamiento & purificación , Bacterias Grampositivas/aislamiento & purificación , Periodontitis Periapical/microbiología , Reacción en Cadena de la Polimerasa , Bacterias Anaerobias/clasificación , Bacterias Anaerobias/genética , Cartilla de ADN/química , ADN Bacteriano/análisis , ADN Ribosómico/análisis , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/genética , Humanos , Periodontitis Periapical/diagnóstico por imagen , Radiografía , Tratamiento del Conducto Radicular
10.
Int J Paediatr Dent ; 14(3): 182-91, 2004 May.
Artículo en Inglés | MEDLINE | ID: mdl-15139953

RESUMEN

OBJECTIVES: The usage of minimally invasive procedures and attention to patient comfort are of great importance, especially for dental treatment in small children. This has led to the development of chemomechanical methods for caries removal. The aim of this study was to compare the efficacy of chemomechanical caries removal with that of conventional excavation in reducing the count of the cariogenic flora. DESIGN AND SETTING: Subjects for this study were chosen from children admitted to dental clinic for restorative procedures under general anaesthesia. SAMPLES AND METHODS: Twenty-one children (mean age 43.5 +/- 12.0 months) with early childhood caries were included in this study. Two primary teeth with comparable degrees of carious destruction were chosen in each child (n = 42) for caries removal with Carisolv' or by means of rotary instruments. Samples from carious dentine were taken with a sterile scraping instrument, then all softened dentine was removed and a second sample was taken. All samples (n = 84) were serially diluted and plated on two different nutrient agar plates. RESULTS: After 24 h of incubation, colony forming units were determined for total bacterial counts and lactobacilli. Twelve per cent of the samples from carious dentine contained more than 10(6) bacteria, 23.8% contained more than 10(5) lactobacilli. Both methods of caries removal produced a statistically significant reduction in the bacterial counts (P = 0.0001). In at least 90.5% of the samples taken after the removal, the total bacterial count was below 10(2), and in 95.2% lactobacilli fell below 10(2). CONCLUSION: These results indicate that the efficacy of chemomechanical removal of carious dentine in children by means of Carisolv' is comparable to the results obtained by conventional methods, and thus might serve as a suitable alternative.


Asunto(s)
Bacterias/efectos de los fármacos , Caries Dental/microbiología , Preparación de la Cavidad Dental/métodos , Ácido Glutámico/uso terapéutico , Leucina/uso terapéutico , Lisina/uso terapéutico , Bacterias/crecimiento & desarrollo , Niño , Preescolar , Recuento de Colonia Microbiana , Caries Dental/terapia , Preparación de la Cavidad Dental/instrumentación , Dentina/microbiología , Humanos , Lactobacillus/efectos de los fármacos , Lactobacillus/crecimiento & desarrollo , Microscopía Electrónica de Rastreo
11.
Eur J Med Res ; 8(1): 25-32, 2003 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-12578751

RESUMEN

BACKGROUND: Immunosuppressants play an essential role in transplantation therapy. In view of the side effects, e.g. gingival overgrowth, the present in vitro study was performed in order to investigate the effect of selected immunosuppressants on metabolic activities of gingival fibroblasts. Furthermore, the effect on the growth of six oral microorganisms was investigated. METHODS: Human gingival fibroblasts were incubated in the presence of azathioprine (Aza), cyclosporin A (CsA), tacrolimus (Tac) or mycophenolatmofetil (Myc). PGE subset 2 release was determined by means of a specific competitive enzyme immunoassay, using monoclonal antibodies specific for PGE subset 2 (clone E2R1). The protein content was measured spectrophotometrically. A redox indicator system was employed to assess the proliferation activity. In an additional trial the growth of six strains of oral bacteria (A. viscosus T14V, S. oralis H1, S. mutans 10449, C. gingivalis DR2001, A. actinomycetemcomitans Y4, and M. micros 33270) in the presence of the immunosuppressants was measured. RESULTS: In comparison with the controls, the PGE subset 2 release was increased by 39.3% following incubation with Aza, and by 77.0% with CsA. The protein concentrations (1 g immunosuppressant / ml medium) were reduced by 26.0% for Aza and 17.0% for Myc. Furthermore, a drug-dependent inhibition in the cell proliferation rate was noted after an incubation period of 6 hours (Aza 70.7%, CsA 78.2%, Myc 69.8%, Tac 64.0%). The most pronounced growth-inhibiting effects were observed for CsA at values ranging from 21.0% (S. mutans 10449) to 48.6% (A. viscosus T14V) growth inhibition. CONCLUSIONS: The present study with common immunsuppresants demonstrated both a medication- and dose-dependent alteration in the metabolic activity of gingival fibroblasts. Furthermore, growth-inhibitory effects on the selected bacterial strains could be observed.


Asunto(s)
Azatioprina/farmacología , Placa Dental/microbiología , Dinoprostona/metabolismo , Fibroblastos/efectos de los fármacos , Encía/citología , Inmunosupresores/farmacología , Actinomyces viscosus/efectos de los fármacos , Actinomyces viscosus/crecimiento & desarrollo , Aggregatibacter actinomycetemcomitans/efectos de los fármacos , Aggregatibacter actinomycetemcomitans/crecimiento & desarrollo , Capnocytophaga/efectos de los fármacos , Capnocytophaga/crecimiento & desarrollo , División Celular/efectos de los fármacos , Células Cultivadas , Ciclosporina/farmacología , Fibroblastos/citología , Fibroblastos/metabolismo , Humanos , Ácido Micofenólico/análogos & derivados , Ácido Micofenólico/farmacología , Peptostreptococcus/efectos de los fármacos , Peptostreptococcus/crecimiento & desarrollo , Biosíntesis de Proteínas , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/crecimiento & desarrollo , Tacrolimus/farmacología
12.
Annu Rev Phytopathol ; 39: 419-60, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-11701872

RESUMEN

This article summarizes studies of viral coat (capsid) proteins (CPs) of RNA plant viruses. In addition, we discuss and seek to interpret the knowledge accumulated to data. CPs are named for their primary function; to encapsidate viral genomic nucleic acids. However, encapsidation is only one feature of an extremely diverse array of structural, functional, and ecological roles played during viral infection and spread. Herein, we consider the evolution of viral CPs and their multitude of interactions with factors encoded by the virus, host plant, or viral vector (biological transmission agent) that influence the infection and epidemiological facets of plant disease. In addition, applications of today's understanding of CPs in the protection of crops from viral infection and use in the manufacture of valuable compounds are considered.


Asunto(s)
Cápside/fisiología , Enfermedades de las Plantas/virología , Virus de Plantas/genética , Virus ARN/fisiología , Biotecnología , Cápside/genética , Genoma Viral , Filogenia , Virus de Plantas/fisiología , Virus ARN/genética , ARN Viral/genética , ARN Viral/fisiología
13.
J Virol ; 75(19): 9538-42, 2001 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11533217

RESUMEN

A Carnation ringspot virus (CRSV) variant (1.26) was identified that accumulates virions but is incapable of forming a systemic infection. The 1.26 capsid protein gene possesses a Ser-->Pro mutation at amino acid 282. Conversion of 1.26 amino acid 282 to Ser restored systemic infection, while the reciprocal mutation in wild-type CRSV abolished systemic infection. Similar mutations introduced into the related Red clover necrotic mosaic virus capsid protein gene failed to induce the packaging but nonsystemic movement phenotype. These results provide additional support for the theory that virion formation is necessary but not sufficient for systemic movement with the dianthoviruses.


Asunto(s)
Cápside/genética , Virus de Plantas/fisiología , Plantas/virología , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Datos de Secuencia Molecular , Mutación Puntual , Virión/fisiología , Virulencia/genética , Ensamble de Virus/genética
14.
Int Dent J ; 50(2): 79-85, 2000 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10945186

RESUMEN

AIM: The aim of the study was to compare the attachment of two typical strains of oral bacteria to four denture base materials. DESIGN: In vitro study. METHOD: Discs of acrylic resin denture base materials (Paladon 65, polished and unpolished; Palapress; Microbase, polished and unpolished, and Triad VLC) were placed into Petri dishes with Schaedler's medium, inoculated with Streptococcus oralis 34 or Actinomyces viscosus T14V. MAIN OUTCOME MEASURES: After 24 h or 48 h the numbers of adhering bacteria were measured. RESULTS: The bacteria adhered to all discs in similar numbers: 3-9 x 10(6)/ml (viable cell count) and 9-22 x 10(8)/ml (total cell count) for T14V, and 2-6 x 10(6)/ml (viable cell count) and 1.5-3 x 10(8)/ml (total cell count) for 34. CONCLUSIONS: Polishing had little effect on adherence. Denture base materials are not resistant against adherence and possible surface damage by oral bacteria. Therefore, thorough oral hygiene is important for denture wearers.


Asunto(s)
Actinomyces viscosus/fisiología , Adhesión Bacteriana , Bases para Dentadura/microbiología , Streptococcus oralis/fisiología , Diente Artificial/microbiología , Resinas Acrílicas , Recuento de Colonia Microbiana , Placa Dental/microbiología , Pulido Dental , Microscopía Electrónica de Rastreo , Propiedades de Superficie
16.
Mol Plant Microbe Interact ; 13(5): 512-9, 2000 May.
Artículo en Inglés | MEDLINE | ID: mdl-10796017

RESUMEN

A novel genetic screen was used to identify host factors in Arabidopsis thaliana that suppress mutations in the Cauliflower mosaic virus (CaMV) movement protein gene (gene I). A series of small mutations was made in gene I and the mutations were tested for their suitability in a suppressor screen. The first round of screening yielded only revertants or second-site mutations in gene I. A derivative of one of the second-site mutant viruses (N7) that was delayed in symptom production was used in a second round of screening for suppressor plants that accelerated symptom production. Two candidate suppressor plants were found that accelerated by 1 to 4 days the first appearance of symptoms caused by the mutant viruses. One of the suppressors (5-2), called asc1 (acceleration of symptoms by CaMV N7), was mapped to chromosome 1. Two additional loci that differentially affect N7 virus susceptibility in the parental Columbia and Ler ecotypes were mapped to chromosomes 3 and 4 by quantitative trait locus (QTL) analysis.


Asunto(s)
Arabidopsis/genética , Caulimovirus/genética , Genes Supresores , Proteínas Virales/genética , Caulimovirus/patogenicidad , Mutagénesis , Proteínas de Movimiento Viral en Plantas , Reacción en Cadena de la Polimerasa , Virulencia
17.
Eur J Med Res ; 5(12): 523-9, 2000 Dec 29.
Artículo en Inglés | MEDLINE | ID: mdl-11147996

RESUMEN

BACKGROUND/AIMS: Clinical studies have shown the efficacy of mouthrinses in reducing plaque accumulation and inflammation of oral tissues. The aim of this in vitro study was to compare the effect of three mouthrinses: Meridol, an organic amine/ stannous fluoride solution; Parodontax, containing herbal ingredients; and an 0.8 % Emser salt solution, on the growth of oral bacteria and dental plaque. METHODS: Growth of Actinomyces viscosus T14V, Capnocytophaga ochracea 25, C. sputigena 4, Actinobacillus actinomycetemcomitans (A.a.) Y4, and pooled supragingival plaque in the presence of the various mouthrinses, applied to paper discs, was tested in an agar diffusion test. In a second series of tests, the 4 bacterial strains were exposed to the agents for about 3 min to simulate rinsing, then the agent was removed, and the bacteria were inoculated into fresh nutrient broth. After 48 h bacterial growth was measured in a spectrophotometer and compared with the controls. RESULTS: In the agar diffusion test only Meridol, the organic amine/stannous fluoride-containing solution, could inhibit bacterial growth, except for A. a. Y4. When the bacteria where in contact with the agents for only a few minutes these results were confirmed. Neither Paradontax nor Emser salt inhibited the growth of the bacteria, and A. a. Y4 proved to be resistant to all three agents. Growth of the other three strains was inhibited by Meridol 92-99% (undiluted), 85-96% (1:5) and 83-98% (1:10). CONCLUSIONS: We conclude that only Meridol contains ingredients capable of inhibiting the growth of oral bacteria in vitro. The efficacy of the other two mouthrinses in reducing plaque accumulation in vivo has to be explained by other mechanisms.


Asunto(s)
Actinomyces/efectos de los fármacos , Aminas/farmacología , Antisépticos Bucales/farmacología , Extractos Vegetales/farmacología , Sales (Química)/farmacología , Bicarbonato de Sodio/farmacología , Fluoruros de Estaño/farmacología , Actinomyces/crecimiento & desarrollo , Agar , Aggregatibacter actinomycetemcomitans/efectos de los fármacos , Aggregatibacter actinomycetemcomitans/crecimiento & desarrollo , Capnocytophaga/efectos de los fármacos , Capnocytophaga/crecimiento & desarrollo , Placa Dental/prevención & control , Combinación de Medicamentos , Técnicas In Vitro , Aguas Minerales , Higiene Bucal , Fitoterapia
18.
Biochem Biophys Res Commun ; 266(1): 203-7, 1999 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-10581190

RESUMEN

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a mammary gland carcinogen present in the human diet in cooked meat. To examine if PhIP and its reactive metabolite N-hydroxy-PhIP inhibit apoptosis in human mammary epithelial MCF-10A cells, confluent cultures deprived of serum and growth factors were incubated for 24 h with either compound. The percentages of dead cells (mean +/- SEM, n = 3) as measured by trypan blue exclusion were 5.7 +/- 0.6, 3.4 +/- 0.3, 2.7 +/- 0.3, and 0.2 +/- 0.003%, in control, 1 microM N-hydroxy-PhIP-, 5 microM N-hydroxy-PhIP-, and 100 microM PhIP-treated dishes, respectively. The expression of Bcl-2 and Bcl-x(L) as quantitated by Western blotting was 1.2- to 1.9-fold higher in the treated groups. PhIP-DNA adducts induced by N-hydroxy-PhIP in MCF-10A cells measured by the (32)P-postlabeling assay were low (<1 x 10(7), relative adduct labeling). No adducts were detected after incubation with PhIP. Western blot analysis indicated that PhIP increased ERK2 phosphorylation concomitant with Bcl-2. The results suggest that the inhibition of cell death in mammary epithelial cells by PhIP occurs independently of PhIP-DNA adducts and may involve enhanced signaling through the MAP kinase pathways.


Asunto(s)
Apoptosis/efectos de los fármacos , Mama/citología , Carcinógenos/farmacología , Células Epiteliales/citología , Calor , Imidazoles/farmacología , Carne , Mama/efectos de los fármacos , Carcinógenos/metabolismo , Línea Celular , Núcleo Celular/efectos de los fármacos , Medio de Cultivo Libre de Suero , Aductos de ADN/análisis , Aductos de ADN/genética , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Imidazoles/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Piridinas/metabolismo , Piridinas/farmacología , Factores de Tiempo , Proteína bcl-X
19.
Carcinogenesis ; 20(7): 1309-14, 1999 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10383905

RESUMEN

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a compound from cooked meat, is an established mammary gland carcinogen in female rats. Four doses of PhIP (150 mg/kg, p.o., once per day) were given to lactating Sprague-Dawley rats separated from their 10-day-old pups to initiate involution of the gland. Twenty-four hours after the last dose, apoptotic index in the mammary gland, as measured by the TUNEL assay, was significantly higher in the gland from control rats than in the PhIP-treated rats (4.757 +/- 1.066 versus 1.905 +/- 0.248%; P < 0.05). In comparison with controls, alveoli in the mammary gland of PhIP-treated rats were also visibly larger and contained more secretory epithelial cells. The expression of Bax, a stimulator of apoptosis, and Bcl-2, an inhibitor of apoptosis, were quantitated by western blotting. Accordingly, Bax expression was 2.7-fold higher in control rats, whereas Bcl-2 expression was 3.1-fold higher in PhIP-treated rats, both changes being statistically different (Student's t-test, P < 0.05). Immunohistochemistry further confirmed a lower expression of Bax and higher expression of Bcl-2 in secretory alveolar epithelial cells of the PhIP-treated mammary gland. The findings are consistent with the notion that exposure to PhIP retarded involution via partial inhibition of programmed cell death. To investigate possible mechanisms for the inhibitory effects of PhIP on mammary gland involution, serum levels of prolactin, an important hormone for the maintenance of lactation, were measured in virgin rats with regular estrous cycles given PhIP (150 mg/kg, p.o.) on the morning of diestrous. After one estrous cycle, on proestrous morning, serum prolactin levels were 1.3-fold higher after PhIP than after control vehicle (one-way ANOVA, Fisher LSD multiple comparison test, P < 0. 05). PhIP exposure during involution was associated with the induction of benign mammary tumors. Seven out of 12 rats developed fibroadenomas, and one developed a tubulopapillary carcinoma within 1 year of receiving PhIP administration during involution (150 mg/kg, p.o., once per day for 5 days), and a high-fat diet (23.5% corn oil). An increase in serum prolactin level and the effects on mammary gland apoptosis seen with PhIP may have implications for the mechanisms of carcinogenic targeting of PhIP to the mammary gland.


Asunto(s)
Imidazoles/farmacología , Lactancia/fisiología , Glándulas Mamarias Animales/efectos de los fármacos , 9,10-Dimetil-1,2-benzantraceno/farmacología , Animales , Apoptosis , Western Blotting , Pruebas de Carcinogenicidad , Carcinógenos/farmacología , Estradiol/sangre , Femenino , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Lactancia/sangre , Lactancia/efectos de los fármacos , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/metabolismo , Glándulas Mamarias Animales/fisiología , Prolactina/sangre , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Radioinmunoensayo , Ratas , Ratas Sprague-Dawley , Proteína X Asociada a bcl-2
20.
Int Dent J ; 49(4): 231-9, 1999 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-10858759

RESUMEN

Three tooth-coloured, resin-based restorative materials (Charisma, Dyract, and Pertac) were exposed to typical oral bacteria (S. mutans, S. oralis and A. naeslundii) over a period of up to 35 days. The three strains of bacteria all colonised the resin-based materials within a few hours and formed thick bacterial films. Determination of the bacterial glucose consumption and lactate production during the incubation period showed no difference from the controls which contained no resin samples. Following the experimental exposure, the materials were examined by scanning electron microscopy (SEM) for possible surface damage and roughness was measured in a perthometer. Little damage to the resin-based composite material surfaces (Charisma, Pertac) could be observed, whereas the polyacid-modified composite material (Dyract) showed greater damage. There was a significant difference in the resin surface roughness after exposure to S. mutans and to A. naeslundii. The study clearly showed that the bacteria used strongly adhered to the resin-based restorative materials. As a consequence of bacterial colonisation and/or poor oral hygiene, damage to the restorative materials might develop. This suggests the need for dentists to evaluate personal oral hygiene, along with general indications and economic factors, in selecting materials for restorations, since the known anti-bacterial properties of amalgam are considerable.


Asunto(s)
Actinomyces/crecimiento & desarrollo , Restauración Dental Permanente , Resinas Sintéticas , Streptococcus mutans/crecimiento & desarrollo , Streptococcus oralis/crecimiento & desarrollo , Actinomyces/metabolismo , Actinomyces/fisiología , Adhesión Bacteriana , Biopelículas/crecimiento & desarrollo , Bisfenol A Glicidil Metacrilato , Distribución de Chi-Cuadrado , Compómeros , Resinas Compuestas , Glucosa/metabolismo , Humanos , Lactatos/metabolismo , Metacrilatos , Microscopía Electrónica de Rastreo , Higiene Bucal , Silicatos , Streptococcus mutans/metabolismo , Streptococcus mutans/fisiología , Streptococcus oralis/metabolismo , Streptococcus oralis/fisiología , Propiedades de Superficie , Factores de Tiempo
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