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1.
Clin Psychol Rev ; 100: 102251, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36716584

RESUMEN

This systematic review aimed to determine whether the use of specific behaviour change technique (BCT) groups are associated with greater effectiveness for psychosocial interventions delivered to family and close friends (FCFs) impacted by addiction. A systematic search of peer-reviewed and grey literature published until August 2021 identified 32 studies in 38 articles. An established BCT taxonomy (93 BCTs clustered into 16 groups) was adapted (inclusion of seven additional BCT groups) and applied to 57 interventions. The meta-analyses indicated that some, but not all, FCF outcomes were improved by the exclusion of BCTs within several groups (Reward and Threat, Scheduled Consequences, Confrontation of the Addicted Person to Engage in Treatment, and Goals and Planning) and inclusion of BCTs within the Restoring a Balanced Lifestyle group. Addicted person outcomes were improved by the inclusion of some BCTs within several groups (Repetition and Substitution, Reward and Threat, Scheduled Consequences, and Restoring a Balanced Lifestyle). Relationship functioning outcomes were improved by the inclusion of BCTs within the Confrontation of the Addicted Person to Engage in Treatment group. Future research involving the development and evaluation of numerous interventions or comprehensive multi-component interventions that can address the various needs of FCFs, without counteracting them, is required.


Asunto(s)
Terapia Conductista , Amigos , Humanos , Terapia Conductista/métodos , Recompensa
2.
Addict Behav ; 109: 106470, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32485545

RESUMEN

BACKGROUND: Caffeine Use Disorder (CUD) is not yet formally recognized in the DSM-5, but emerging evidence suggests CUD could impact up to one in five people. The primary aim of this study was to estimate levels of caffeine consumption and its associations with CUD and withdrawal, taking socio-demographic characteristics (age, gender, ethnicity, income) into account. Secondary aims were to measure caffeine-related harm and treatment preferences. METHODS: We administered an online cross-sectional survey via Facebook to a convenience sample of 2379 adults in New Zealand. Caffeine consumption was assessed across six products: coffee, tea, energy drinks, cola, alcohol mixed with caffeine, and other products (e.g., caffeine pills, sports supplements). RESULTS: 20% of participants met proposed criteria for CUD, with 30% meeting DSM-5 criteria for caffeine withdrawal. Moderate (200-400 mg per day) and high consumption (>400 mg per day) was associated with a three-fold increase in the odds of CUD and a two-fold increase in the odds of withdrawal. Women and current smokers were at higher risk of CUD and withdrawal even at moderate consumption levels. Nearly 85% of respondents experienced at least one caffeine-related harm in the past 12-months. The number of harms increased with level of caffeine consumption. Nearly 50% indicated a self-help treatment for caffeine reduction would be of interest. CONCLUSIONS: High rates of CUD and caffeine withdrawal amongst moderate caffeine users, women and smokers suggests caffeine consumption guidelines may need refinement. Caffeine-related harm that is not clinically meaningful may still be of concern to individuals and warrants further investigation.


Asunto(s)
Cafeína , Bebidas Energéticas , Adulto , Cafeína/efectos adversos , Café , Estudios Transversales , Bebidas Energéticas/efectos adversos , Femenino , Humanos , Nueva Zelanda/epidemiología , Encuestas y Cuestionarios
3.
Addict Behav ; 103: 106257, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-31884377

RESUMEN

BACKGROUND: The Problem Gambling Severity Index (PGSI) was intended for use in epidemiological research with gamblers across the continuum of risk. Its utility within clinical settings, where the majority of clients are problem gamblers, has been brought into question. AIMS: (1) Identify refined categories for the problem gambling category of the PGSI in help-seeking gamblers; (2) Validate these categories using the Gambling Symptom Assessment Scale (G-SAS); (3) Explore the relationship of these categories with indices of gambling and help-seeking behaviour. METHODS: Secondary data analysis of help-seeking problem gamblers from the Australian online gambling counselling/support service (Gambling Help Online [GHO]) from October 2012 to December 2015 (n = 5,881) and trial data evaluating an Australian online self-directed program for gambling (GamblingLess; n = 198). Both datasets included the PGSI, gambling frequency and expenditure. The GamblingLess dataset also included the G-SAS and help-seeking behaviour. RESULTS: A Latent Class Analysis, using GHO data, identified a 2-class solution. Multiple analytical methods identified a cut-off value of ≥ 19 distinguishing this 2-class solution (low problem severity: Median = 16; high problem severity: Median = 23). High problem severity gamblers had increased odds of being categorised in the higher GSAS category, greater gambling expenditure and having sought face-to-face support. The refined categories were not associated with gambling frequency, distance-based or self-directed help-seeking. CONCLUSION: These findings are consistent with a stepped-care approach, whereby individuals with higher severity may be better suited to more intensive interventions and individuals with lower severity could commence with less intensive interventions and step-up to intensive interventions.


Asunto(s)
Conducta Adictiva/psicología , Juego de Azar/psicología , Psicometría/instrumentación , Índice de Severidad de la Enfermedad , Adolescente , Adulto , Australia , Femenino , Conducta de Búsqueda de Ayuda , Humanos , Análisis de Clases Latentes , Masculino , Reproducibilidad de los Resultados , Adulto Joven
4.
Clin Psychol Rev ; 74: 101784, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31759246

RESUMEN

Non-gambling specialist services, such as primary care, alcohol and other drug use, and mental health services, are well placed to enhance the identification of people with gambling problems and offer appropriate generalist first level interventions or referral. Given time and resource demands, many of these clinical services may only have the capacity to administer very short screening instruments. This systematic review was conducted to provide a resource for health service providers and researchers in identifying the most accurate brief (1-5 item) screening instruments to identify problem and at-risk gambling for their specific purposes and populations. A systematic search of peer-reviewed and grey literature from 1990 to 2019 identified 25 articles for inclusion. Meta-analysis revealed five of the 20 available instruments met criteria for satisfactory diagnostic accuracy in detecting both problem and at-risk gambling: Brief Problem Gambling Screen (BPGS-2), NODS-CLiP, Problem Gambling Severity Index-Short Form (PGSI-SF), NODS-PERC, and NODS-CLiP2. Of these, the NODS-CLiP and NODS-PERC have the largest volume of diagnostic data. The Lie/Bet Questionnaire and One-Item Screen are also promising shorter options. Because these conclusions are drawn from a relatively limited evidence base, future studies evaluating the diagnostic accuracy of existing brief instruments across settings, age groups, and timeframes are needed.


Asunto(s)
Juego de Azar/diagnóstico , Escalas de Valoración Psiquiátrica/normas , Humanos
5.
Internet Interv ; 18: 100264, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31890617

RESUMEN

INTRODUCTION: Countries with legalized gambling offer a network of government funded face-to-face therapy, but usage of this expertise is on the decline. One way to address this issue is to recruit therapists from existing services whereby they provide guidance for the delivery of internet delivered CBT. OBJECTIVE: To explore the experiences and perceptions of therapists supporting guided online cognitive-behavioural therapy. METHODS: Interviewees were a sub-sample of therapists from a randomised trial that investigated the relative efficacy of online guided self-directed versus pure self-directed interventions in Australia. RESULTS: In-person, semi-structured interviews with seven service providers were completed, and thematic content analysis identified five themes which related to: participant suitability and screening (e.g., motivation, computer literacy and access); program content and modality acceptability (e.g., amount of content, look and feel); participant information and management (e.g., program engagement and progression); email communication (e.g., use of templates, appointments, rapport building), and; ongoing service integration (e.g., infrastructure, confidence in product). Overall experiences and perceptions of therapists were positive, notwithstanding barriers concerning assessment of participant suitability, participant management systems and low participant engagement. CONCLUSIONS: Key themes emphasized the benefits of Internet-based interventions for problem gambling, and suggested several areas for improvement. Results should inform the development of future treatments to enable flexible tailoring of interventions to individuals.

6.
Addict Behav ; 92: 32-37, 2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-30579115

RESUMEN

Addressing gambling problems across a continuum requires understanding of low severity problems, as well as severe levels of problem gambling or disorder. The aims of this study were thus to derive a map of how problematic gambling behaviours and harms are situated across a continuum, and identify the best available indicators of low severity problems to inform assessment and secondary prevention. This involved the Rasch analyses of baseline data from the Quinte Longitudinal Study (QLS); a community-based survey involving random-digit dialling of numbers around Belleville, Canada. Participants were n = 1305 adults with non-zero scores across 26-items from: the Problem Gambling Severity Index (PGSI); the NORC DSM Screen for Gambling Problems (NODS); and the Problem and Pathological Gambling Measure (PPGM). Results indicated that item-level measures except chasing losses provided fit to the Rasch model, and most were clustered within a narrow region of the continuum which resembled addictive disorders. At the most severe end were mainly items about harms, while there were few items representing low severity levels (feeling guilty, betting more than one can afford, attempts to reduce gambling, gambling more than intended). There was Differential Item Functioning (DIF) for several indicators of low severity problems. The findings suggest that measures remain closely aligned with psychiatric models and are suited for discriminating across severe levels of problem gambling or addictive disorder. Although cognitive-affective and behavioural indicators comprise the best available indicators of low severity symptoms, there is an urgent need for improvements in conceptualisation and measurement.


Asunto(s)
Conducta Adictiva/epidemiología , Juego de Azar/epidemiología , Adolescente , Adulto , Canadá/epidemiología , Femenino , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Psicometría , Índice de Severidad de la Enfermedad , Adulto Joven
7.
Addict Behav ; 87: 272-275, 2018 12.
Artículo en Inglés | MEDLINE | ID: mdl-29935737

RESUMEN

Research investigating rates of help-seeking for problem gambling has traditionally focused on the uptake of face-to-face gambling services alone, despite the World Health Organisation defining help-seeking as any action or activity undertaken to improve or resolve emotional, psychological or behavioural problems. The primary aim of this study is to examine the full range of help-seeking options utilised by gamblers, and to determine whether administering a comprehensive list of help options yields higher help-seeking rates than a single item measure. A one-item and expanded 14-item help-seeking Questionnaire (the Help-Seeking Questionnaire; HSQ) were administered to 277 problem gamblers seeking help online. We found the 14-item HSQ yielded a significantly higher level of lifetime professional help-seeking (70%) compared to the one-item measure (22%). When we included self-directed activities, 93% of gamblers reported they had previously attempted at least one activity to reduce their gambling. Current measurement of help-seeking appears to underestimate the range of activities currently undertaken by gamblers to reduce their gambling. Surveys need to include the one-item HSQ (over the past 12 months have you sought professional help or advice (online, by phone, or in person), support from family or friends, or did something by yourself to limit or reduce your gambling?) or the three-item HSQ which measures engagement of face-to-face services (i.e., counselling, advice, groups), distance-based (i.e., anonymous telephone, online) and self-directed (i.e., activities not involving professional oversight) activities separately. The full 14-item screen can be administered when brief screens are positive to ensure accurate measurement of help-seeking.


Asunto(s)
Juego de Azar/psicología , Conducta de Búsqueda de Ayuda , Autonomía Personal , Adolescente , Adulto , Anciano , Australia , Consejo/estadística & datos numéricos , Femenino , Humanos , Internet/estadística & datos numéricos , Masculino , Persona de Mediana Edad , Autoeficacia , Adulto Joven
8.
J Gambl Stud ; 34(1): 307-320, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28884260

RESUMEN

Despite high rates of comorbidity between problem gambling and mental health disorders, few studies have examined barriers or facilitators to the implementation of screening for problem gambling in mental health services. This exploratory qualitative study identified key themes associated with screening in mental health services. Semi-structured interviews were undertaken with 30 clinicians and managers from 11 mental health services in Victoria, Australia. Major themes and subthemes were identified using qualitative content analysis. Six themes emerged including competing priorities, importance of routine screening, access to appropriate screening tools, resources, patient responsiveness and workforce development. Barriers to screening included a focus on immediate risk as well as gambling being often considered as a longer-term concern. Clinicians perceived problem gambling as a relatively rare condition, but did acknowledge the need for brief screening. Facilitators to screening were changes to system processes, such as identification of an appropriate brief screening instrument, mandating its use as part of routine screening, as well as funded workforce development activities in the identification and management of problem gambling.


Asunto(s)
Conducta Adictiva/terapia , Juego de Azar/terapia , Servicios de Salud Mental/estadística & datos numéricos , Aceptación de la Atención de Salud/psicología , Conducta Adictiva/psicología , Comorbilidad , Femenino , Juego de Azar/psicología , Humanos , Masculino , Investigación Cualitativa , Victoria/epidemiología
9.
BMJ Open ; 7(3): e013490, 2017 03 02.
Artículo en Inglés | MEDLINE | ID: mdl-28255094

RESUMEN

INTRODUCTION: The primary purpose of this study is to evaluate the relative effectiveness of 2 of the best developed and most promising forms of therapy for problem gambling, namely face-to-face motivational interviewing (MI) combined with a self-instruction booklet (W) and follow-up telephone booster sessions (B; MI+W+B) and face-to-face cognitive-behavioural therapy (CBT). METHODS AND ANALYSIS: This project is a single-blind pragmatic randomised clinical trial of 2 interventions, with and without the addition of relapse-prevention text messages. Trial assessments take place pretreatment, at 3 and 12 months. A total of 300 participants will be recruited through a community treatment agency that provides services across New Zealand and randomised to up to 10 face-to-face sessions of CBT or 1 face-to-face session of MI+W+up to 5 B. Participants will also be randomised to 9 months of postcare text messaging. Eligibility criteria include a self-perception of having a current gambling problem and a willingness to participate in all components of the study (eg, read workbook). The statistical analysis will use an intent-to-treat approach. Primary outcome measures are days spent gambling and amount of money spent per day gambling in the prior month. Secondary outcome measures include problem gambling severity, gambling urges, gambling cognitions, mood, alcohol, drug use, tobacco, psychological distress, quality of life, health status and direct and indirect costs associated with treatment. ETHICS AND DISSEMINATION: The research methods to be used in this study have been approved by the Ministry of Health, Health and Disability Ethics Committees (HDEC) 15/CEN/99. The investigators will provide annual reports to the HDEC and report any adverse events to this committee. Amendments will also be submitted to this committee. The results of this trial will be submitted for publication in peer-reviewed journals and as a report to the funding body. Additionally, the results will be presented at national and international conferences. TRIAL REGISTRATION NUMBER: ACTRN12615000637549.


Asunto(s)
Conducta Adictiva/terapia , Terapia Cognitivo-Conductual/métodos , Juego de Azar/terapia , Entrevista Motivacional , Folletos , Envío de Mensajes de Texto , Femenino , Humanos , Masculino , Proyectos de Investigación , Método Simple Ciego , Teléfono
10.
BMJ Open ; 7(2): e014226, 2017 02 23.
Artículo en Inglés | MEDLINE | ID: mdl-28235970

RESUMEN

INTRODUCTION: The prevalence of disordered gambling worldwide has been estimated at 2.3%. Only a small minority of disordered gamblers seek specialist face-to-face treatment, and so a need for alternative treatment delivery models that capitalise on advances in communication technology, and use self-directed activity that can complement existing services has been identified. As such, the primary aim of this study is to evaluate an online self-directed cognitive-behavioural programme for disordered gambling (GamblingLess: For Life). METHODS AND ANALYSIS: The study will be a 2-arm, parallel group, pragmatic randomised trial. Participants will be randomly allocated to a pure self-directed (PSD) or guided self-directed (GSD) intervention. Participants in both groups will be asked to work through the 4 modules of the GamblingLess programme over 8 weeks. Participants in the GSD intervention will also receive weekly emails of guidance and support from a gambling counsellor. A total of 200 participants will be recruited. Participants will be eligible if they reside in Australia, are aged 18 years and over, have access to the internet, have adequate knowledge of the English language, are seeking help for their own gambling problems and are willing to take part in the intervention and associated assessments. Assessments will be conducted at preintervention, and at 2, 3 and 12 months from preintervention. The primary outcome is gambling severity, assessed using the Gambling Symptom Assessment Scale. Secondary outcomes include gambling frequency, gambling expenditure, psychological distress, quality of life and additional help-seeking. Qualitative interviews will also be conducted with a subsample of participants and the Guides (counsellors). ETHICS AND DISSEMINATION: The study has been approved by the Deakin University Human Research and Eastern Health Human Research Ethics Committees. Findings will be disseminated via report, peer-reviewed publications and conference presentations. TRIAL REGISTRATION NUMBER: ACTRN12615000864527; results.


Asunto(s)
Terapia Cognitivo-Conductual/métodos , Juego de Azar/psicología , Juego de Azar/terapia , Adolescente , Adulto , Anciano , Australia , Femenino , Humanos , Internet/estadística & datos numéricos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Calidad de Vida , Proyectos de Investigación , Adulto Joven
11.
J Gambl Stud ; 33(1): 283-299, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27562272

RESUMEN

Research suggests online interventions can have instant impact, however this is yet to be tested with help-seeking adults and in particular those with problem gambling. This study seeks to determine the immediate impact of a single session web-based intervention for problem gambling, and to examine whether sessions evaluated positively by clients are associated with greater improvement. The current study involved 229 participants classified as problem gamblers who agreed to participate after accessing Gambling Help Online between November 2010 and February 2012. Almost half were aged under 35 years of age (45 %), male (57 %) as well as first time treatment seekers (62 %). Participants completed measures of readiness to change and distress both prior to and post-counselling. Following the provision of a single-session of counselling, participants completed ratings of the character of the session (i.e., degree of depth and smoothness) post-counselling. A significant increase in confidence to resist and urge to gamble and a significant decrease in distress (moderate effect size; d = .56 and .63 respectively) was observed after receiving online counselling. A hierarchical regression indicated the character of the session was a significant predictor of change in confidence, however only the sub-scale smoothness was a significant predictor of change in distress. This was the case even after controlling for pre-session distress, session word count and client characteristics (gender, age, preferred gambling activity, preferred mode of gambling, gambling severity, and preferred mode of help-seeking). These findings suggest that single session web-based counselling for problem gambling can have immediate benefits, although further research is required to examine the impact on longer-term outcomes.


Asunto(s)
Consejo/métodos , Juego de Azar/psicología , Juego de Azar/terapia , Terapia Asistida por Computador/métodos , Adulto , Femenino , Conducta de Búsqueda de Ayuda , Humanos , Internet , Masculino , Procesos Mentales , Persona de Mediana Edad , Autoimagen
12.
Clin Oncol (R Coll Radiol) ; 27(7): 382-6, 2015 07.
Artículo en Inglés | MEDLINE | ID: mdl-25805312

RESUMEN

AIMS: To examine the relationship between post-implant computed tomography dosimetry and long-term prostate-specific antigen relapse-free survival in patients treated with iodine 125 (I-125) low dose rate prostate brachytherapy as monotherapy and, second, to audit recent practice against Royal College of Radiologists' (RCR) guidelines after the re-introduction of post-implant dosimetry for all patients in our centre. MATERIALS AND METHODS: Between March 1995 and September 2007, 2157 consecutive patients with localised prostate cancer underwent I-125 permanent prostate brachytherapy as monotherapy in a single UK centre. All patients were transrectal ultrasound planned delivering a 145 Gy (TG 43) minimum peripheral dose. None received supplemental external beam radiotherapy. Post-implant computed tomography-based dosimetry was undertaken between 4 and 6 weeks after treatment and was available for 711 (33%). Outcomes were analysed in terms of the relationship of D90 to prostate-specific antigen relapse-free survival (nadir 2+ definition) and all patients had a minimum follow-up of 5 years. For contemporary patients from 2011, quality metrics from post-implant computed tomography as defined by RCR guidelines are presented. RESULTS: A mean D90 of 138.7 Gy (standard deviation 24.7) was achieved for the historic cohort. Biochemical control at 10 years was 76% in patients with D90 > 140 Gy and 68% in those with D90 < 140 Gy (P < 0.01). In current practice, over the last 3 years the mean (standard deviation) D90 has increased from 154 (15.3) Gy in 2011 to 164 (13.5) Gy in 2013. Similarly, an increase in the mean (standard deviation) V100 from 92 (4.4) to 95 (3.2) % is noted over time. No difference between clinicians was noted. CONCLUSION: D90 values of less than 140 Gy continue to be predictive of increased risk of recurrence of prostate cancer across risk groups with longer follow-up. Quality assurance can be used to ensure improved and consistent implant quality in a team with multiple clinicians.


Asunto(s)
Braquiterapia/métodos , Neoplasias de la Próstata/radioterapia , Adulto , Anciano , Anciano de 80 o más Años , Humanos , Radioisótopos de Yodo/uso terapéutico , Masculino , Persona de Mediana Edad , Antígeno Prostático Específico/sangre , Neoplasias de la Próstata/sangre , Neoplasias de la Próstata/diagnóstico por imagen , Garantía de la Calidad de Atención de Salud , Dosificación Radioterapéutica , Tomografía Computarizada por Rayos X
13.
Addiction ; 110(3): 494-501, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25393315

RESUMEN

BACKGROUND AND AIMS: Problem gamblers are not a homogeneous group and recent data suggest that subtyping can improve treatment outcomes. This study administered three readiness rulers and aimed to identify subtypes of gamblers accessing a national web-based counselling service based on these rulers. METHODS: Participants were 1204 gamblers (99.4% problem gamblers) who accessed a single session of web-based counselling in Australia. Measures included three readiness rulers (importance, readiness and confidence to resist an urge to gamble), demographics and the Problem Gambling Severity Index (PGSI). RESULTS: Gamblers reported high importance of change [mean = 9.2, standard deviation (SD) = 1.51] and readiness to change (mean = 8.86, SD = 1.84), but lower confidence to resist an urge to gamble (mean = 3.93, SD = 2.44) compared with importance and readiness. The statistical fit indices of a latent class analysis identified a four-class model. Subtype 1 was characterized by a very high readiness to change and very low confidence to resist an urge to gamble (n = 662, 55.0%) and subtype 2 reported high readiness and low confidence (n = 358, 29.7%). Subtype 3 reported moderate ratings on all three rulers (n = 139, 11.6%) and subtype 4 reported high importance of change but low readiness and confidence (n = 45, 3.7%). A multinomial logistic regression indicated that subtypes differed by gender (P < 0.001), age (P = 0.01), gambling activity (P < 0.05), preferred mode of gambling (P < 0.001) and PGSI score (P < 0.001). CONCLUSIONS: Problem gamblers in Australia who seek web-based counselling comprise four distinct subgroups based on self-reported levels of readiness to change, confidence to resist the urge to gamble and importance of change.


Asunto(s)
Conducta Adictiva/psicología , Juego de Azar/psicología , Conducta de Búsqueda de Ayuda , Motivación , Adulto , Australia , Conducta Adictiva/clasificación , Conducta Adictiva/rehabilitación , Consejo , Femenino , Juego de Azar/clasificación , Juego de Azar/rehabilitación , Humanos , Internet , Modelos Logísticos , Masculino , Persona de Mediana Edad , Autoinforme , Terapia Asistida por Computador , Resultado del Tratamiento , Adulto Joven
15.
J Biol Chem ; 276(5): 3324-32, 2001 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-11073954

RESUMEN

CP2-related proteins comprise a family of DNA-binding transcription factors that are generally activators of transcription and expressed ubiquitously. We reported a differential display polymerase chain reaction fragment, Psc2, which was expressed in a regulated fashion in mouse pluripotent cells in vitro and in vivo. Here, we report further characterization of the Psc2 cDNA and function. The Psc2 cDNA contained an open reading frame homologous to CP2 family proteins. Regions implicated in DNA binding and oligomeric complex formation, but not transcription activation, were conserved. Psc2 expression in vivo during embryogenesis and in the adult mouse demonstrated tight spatial and temporal regulation, with the highest levels of expression in the epithelial lining of distal convoluted tubules in embryonic and adult kidneys. Functional analysis demonstrated that PSC2 repressed transcription 2.5-15-fold when bound to a heterologous promoter in ES, 293T, and COS-1 cells. The N-terminal 52 amino acids of PSC2 were shown to be necessary and sufficient for this activity and did not share obvious homology with reported repressor motifs. These results represent the first report of a CP2 family member that is expressed in a developmentally regulated fashion in vivo and that acts as a direct repressor of transcription. Accordingly, the protein has been named CP2-Related Transcriptional Repressor-1 (CRTR-1).


Asunto(s)
Proteínas de Unión al ADN/química , Regulación del Desarrollo de la Expresión Génica , Proteínas Represoras/genética , Factores de Transcripción/química , Secuencia de Aminoácidos , Animales , ADN Complementario/análisis , ADN Complementario/aislamiento & purificación , Humanos , Túbulos Renales Distales/metabolismo , Ratones , Datos de Secuencia Molecular , Proteínas de Unión al ARN , Proteínas Represoras/química , Análisis de Secuencia de Proteína , Homología de Secuencia de Aminoácido
16.
Curr Protoc Protein Sci ; Chapter 18: Unit18.2, 2001 May.
Artículo en Inglés | MEDLINE | ID: mdl-18429139

RESUMEN

This unit covers the strategy of the multiple peptide approach to biological scanning, the synthetic protocols, and the handling of peptides after synthesis: cleavage, preliminary purification, storage, and analysis. It is specific for the multipin technique using equipment obtained from Chiron Technologies, although some of the approaches are applicable to other multiple synthesis techniques. This unit also includes protocols for preparing Fmoc-amino acid solutions and for acetylating or biotinylating synthesized peptides.


Asunto(s)
Péptidos/química , Péptidos/síntesis química , Plásticos , Acetilación , Biotinilación , Péptidos/aislamiento & purificación , Tecnología/instrumentación , Tecnología/métodos
17.
J Immunol ; 159(1): 335-43, 1997 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-9200471

RESUMEN

The 10-kDa protein Ag of Mycobacterium leprae, a human GroES hsp10 cognate, is a major T cell Ag in human leprosy infection. We investigated the mechanism for T cell responsiveness to this Ag according to the trimolecular interaction between T cell, peptide, and Ag-presenting element. This research was accomplished by mapping T cell epitopes in leprosy patients and correlating these responses with peptide-MHC binding affinities. We found that the majority of tuberculoid leprosy patients responded to peptides corresponding to residues 25-39 and 28-42. Truncation analysis of these peptides mapped the exact epitope to be within the overlapping region comprising residues 28-39. Responsiveness was correlated with the HLA-DRB5*0101 allele, which bound the peptides with moderate affinity. This allele is linked to HLA-DR2, which is associated with the resistant form of leprosy. Therefore, T cell responsiveness in tuberculoid leprosy may be mediated by the ability of HLA-DRB5*0101 to bind and present peptides of the immunodominant 10-kDa Ag.


Asunto(s)
Chaperonina 10/inmunología , Epítopos de Linfocito T/inmunología , Mycobacterium leprae/inmunología , Linfocitos T/inmunología , Alelos , Secuencia de Aminoácidos , Antígenos Bacterianos/inmunología , Chaperonina 10/genética , Células Clonales , Antígenos HLA-DR/genética , Antígenos HLA-DR/inmunología , Cadenas HLA-DRB5 , Humanos , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido
18.
Infect Immun ; 65(3): 1061-70, 1997 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-9038316

RESUMEN

The mycobacterial 70-kDa heat shock protein (Hsp70) is a dominant antigen during the human T-cell response to mycobacterial infection despite the conserved sequence with the human homolog. To determine whether this response is pathogen specific, CD4+ T-cell clones were isolated from Mycobacterium leprae Hsp70-reactive individuals. The cytokine profile of the clones was mixed, with all of the clones releasing interferon gamma and half releasing interleukin-4 on stimulation, while six demonstrated cytolytic activity. Five clones reacted with the N-terminal half of the molecule, and the epitopes identified were mycobacterium specific. Residues 241 to 260 were identified by three clones, one of which was restricted by HLA-DR7 (DR7), while a DR1-restricted clone identified residues 71 to 90 and residues 261 to 280 were recognized in the context of DR3. The remaining five T-cell clones reacted with the C-terminal half of the molecule, and the precise position of these epitopes was mapped with 12-mer peptides overlapping by 11 residues. Two of these clones identified overlapping epitopes from residues 411 to 425 and 412 to 428, the latter restricted by DR3. Further epitopes were mapped to residues 298 to 313 restricted by DRw53, residues 388 to 406 restricted by DRw52 or DQ2, and residues 471 to 486 restricted by DR1. The sequences of three epitopes, residues 411 to 425, 412 to 428, and 471 to 486, showed significant identity with the equivalent regions of the prototype human Hsp70. However, when amino acid substitutions that made the sequence more like the human sequence were introduced, the changes were tolerated poorly as measured by proliferation, cytokine production, and cytotoxic potential. Therefore, T-cell recognition of the M. leprae Hsp70 antigen occurs in the context of multiple HLA-DR phenotypes and is exquisitely species specific.


Asunto(s)
Proteínas Bacterianas/inmunología , Epítopos , Proteínas HSP70 de Choque Térmico/inmunología , Mycobacterium leprae/inmunología , Linfocitos T/inmunología , Secuencia de Aminoácidos , Células Clonales , Reacciones Cruzadas , Antígenos HLA-DR/análisis , Humanos , Inmunofenotipificación , Datos de Secuencia Molecular
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