971 resultados para Clinical-samples


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The drinking refusal self-efficacy questionnaire (DRSEQ: Young, R.M., Oei, T.P.S., 1996. Drinking expectancy profile: test manual. Behaviour Research and Therapy Centre, University of Queensland, Australia Young, R.M., Oei, T.P.S., Crook, G.M., 1991. Development of a drinking refusal self-efficacy questionnaire. J. Psychopathol. Behav. Assess., 13, 1-15) assesses a person's belief in their ability to resist alcohol. The DRSEQ is a sound psychometric instrument based on exploratory factor analyses, but has not been subjected to confirmatory factor analysis. In total 2773 participants were used to confirm the factor structure of the DRSEQ. Initial analyses revealed that the original structure was not confirmed in the current study. Subsequent analyses resulted in a revised factor structure (DRSEQ-R) being confirmed in community, student and clinical samples. The DRSEQ-R was also found to have good construct and concurrent validity. The factor structure of the DRSEQ-R is more stable than the original structure of the DRSEQ and the revised scale has considerable potential in future alcohol-related research. (c) 2004 Elsevier Ireland Ltd. All rights reserved.

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Antigenic variation in Plasmodium falciparum erythrocyte membrane protein 1, caused by a switch in transcription of the encoding var gene, is an important feature of malaria. In this study, we quantified the relative abundance of var gene transcripts present in P. falciparum parasite clones using real-time reverse transcription-polymerase chain reaction (RT-PCR) and conventional RT-PCR combined with cloning and sequencing, with the aim of directly comparing the results obtained. When there was sufficient abundance of RNA for the real-time RT-PCR assay to be operating within the region of good reproducibility, RT-PCR and real-time RT-PCR tended to identify the same dominant transcript, although some transcript-specific issues were identified. When there were differences in the estimated relative amounts of minor transcripts, the RT-PCR assay tended to produce higher estimates than real-time RT-PCR. These results provide valuable information comparing RT-PCR and real-time RT-PCR analysis of samples with small quantities of RNA as might be expected in the analysis of field or clinical samples.

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We describe the development of a capture enzyme-linked immunosorbent assay for the detection of the dengue virus nonstructural protein NS1. The assay employs rabbit polyclonal and monoclonal antibodies as the capture and detection antibodies, respectively. Immunoaffinity-purified NS1 derived from dengue 2 virus-infected cells was used as a standard to establish a detection sensitivity of approximately 4 ng/ml for an assay employing monoclonal antibodies recognizing a dengue 2 serotype-specific epitope. A number of serotype cross-reactive monoclonal antibodies were also shown to be suitable probes for the detection of NS1 expressed by the remaining three dengue virus serotypes. Examination of clinical samples demonstrated that the assay was able to detect NS1 with minimal interference from serum components at the test dilutions routinely used, suggesting that it could form the basis of a useful additional diagnostic test for dengue virus infection. Furthermore, quantitation of NS1 levels in patient sera may prove to be a valuable surrogate marker for viremia. Surprisingly high levels of NS1, as much as 15 mu g/ml, were found in acute-phase sera taken hom some of the patients experiencing serologically confirmed dengue 2 virus secondary infections but was not detected in the convalescent sera of these patients. In contrast, NS1 could not be detected in either acute-phase or convalescent serum samples taken from patients with serologically confirmed primary infection. The presence of high levels of secreted NS1 in the sera of patients experiencing secondary dengue virus infections, and in the context of an anamnestic antibody response, suggests that NS1 may contribute significantly to the formation of the circulating immune complexes that are suspected to play an important role in the pathogenesis of severe dengue disease.

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In inflammatory diseases, release of oxidants leads to oxidative damage to biomolecules. HOCl (hypochlorous acid), released by the myeloperoxidase/H2O2/Cl- system, can cause formation of phospholipid chlorohydrins, or alpha-chloro-fatty aldehydes from plasmalogens. It can attack several amino acid residues in proteins, causing post-translational oxidative modifications of proteins, but the formation of 3-chlorotyrosine is one of the most stable markers of HOCl-induced damage. Soft-ionization MS has proved invaluable for detecting the occurrence of oxidative modifications to both phospholipids and proteins, and characterizing the products generated by HOCl-induced attack. For both phospholipids and proteins, the application of advanced mass spectrometric methods such as product or precursor ion scanning and neutral loss analysis can yield information both about the specific nature of the oxidative modification and the biomolecule modified. The ideal is to be able to apply these methods to complex biological or clinical samples, to determine the site-specific modifications of particular cellular components. This is important for understanding disease mechanisms and offers potential for development of novel biomarkers of inflammatory diseases. In the present paper, we review some of the progress that has been made towards this goal.

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AIMS: To investigate multiple techniques for the preparation of solid tissue for polymerase chain reaction (PCR) analysis, and to identify the most simple techniques for routine use in the laboratory. METHODS: Techniques for the preparation of arterial tissue samples including homogenisation, ultrafiltration, and treatments involving proteinase K, Gene Clean, lectin, and Fe3+ specific chelators were evaluated using the PCR to amplify both Chlamydia pneumoniae and human DNA. RESULTS: Treatment with either Gene-Clean or lectin and the Fe3+ specific chelator deferoxamine mesylate removed PCR inhibitors from tissue homogenates. Homogenisation followed by GeneClean treatment resulted in the amplification of C pneumoniae DNA from within a section of atherosclerotic carotid artery, implying that C pneumoniae elementary bodies had been disrupted. In eight further clinical samples from patients not known to have C pneumoniae infection, human DNA was amplified and no cross contamination was observed between samples. These samples contained no evidence of C pneumoniae by PCR. CONCLUSIONS: A simple preparation of solid tissue for PCR analysis, involving homogenisation followed by GeneClean treatment has been developed, and is effective for the amplification of both C pneumoniae and human DNA.

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Although the existence of halogenated lipids in lower organisms has been known for many years, it is only since the 1990s that interest in their occurrence in mammalian systems has developed. Chlorinated (and other halogenated) lipids can arise from oxidation by hypohalous acids, such as HOCl, which are products of the phagocytic enzyme myeloperoxidase and are generated during inflammation. The major species of chlorinated lipids investigated to date are chlorinated sterols, fatty acid and phospholipid chlorohydrins, and a-chloro fatty aldehydes. While all of these chlorinated lipids have been shown to be produced in model systems from lipoproteins to cells subjected to oxidative stress, as yet only a-chloro fatty aldehydes, such as 2-chlorohexadecanal, have been detected in clinical samples or animal models of disease. a-Chloro fatty aldehydes and chlorohydrins have been found to have a number of potentially pro-inflammatory effects ranging from toxicity to inhibition of nitric oxide synthesis and upregulation of vascular adhesion molecules. Thus evidence is building for a role of chlorinated lipids in inflammatory disease, although much more research is required to establish the contributions of specific compounds in different disease pathologies. Preventing chlorinated lipid formation and indeed other HOCl-induced damage, via the inhibition of myeloperoxidase, is an area of growing interest and may lead in the future to antimyeloperoxidase-based antiinflammatory therapy. However, other chlorinated lipids, such as punaglandins, have beneficial effects that could offer novel therapies for cancer.

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Significance: Oxidized phospholipids are now well-recognized as markers of biological oxidative stress and bioactive molecules with both pro-inflammatory and anti-inflammatory effects. While analytical methods continue to be developed for studies of generic lipid oxidation, mass spectrometry (MS) has underpinned the advances in knowledge of specific oxidized phospholipids by allowing their identification and characterization, and is responsible for the expansion of oxidative lipidomics. Recent Advances: Studies of oxidized phospholipids in biological samples, both from animal models and clinical samples, have been facilitated by the recent improvements in MS, especially targeted routines that depend on the fragmentation pattern of the parent molecular ion and improved resolution and mass accuracy. MS can be used to identify selectively individual compounds or groups of compounds with common features, which greatly improves the sensitivity and specificity of detection. Application of these methods have enabled important advances in understanding the mechanisms of inflammatory diseases such as atherosclerosis, steatohepatitis, leprosy and cystic fibrosis, and offer potential for developing biomarkers of molecular aspects of the diseases. Critical Issues and Future Directions: The future in this field will depend on development of improved MS technologies, such as ion mobility, novel enrichment methods and databases and software for data analysis, owing to the very large amount of data generated in these experiments. Imaging of oxidized phospholipids in tissue MS is an additional exciting direction emerging that can be expected to advance understanding of physiology and disease.

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Schistosomiasis is a chronically debilitating helminth infection with a significant socio-economic and public health impact. Accurate diagnostics play a pivotal role in achieving current schistosomiasis control and elimination goals. However, many of the current diagnostic procedures, which rely on detection of schistosome eggs, have major limitations including lack of accuracy and the inability to detect pre-patent infections. DNA-based detection methods provide a viable alternative to the current tests commonly used for schistosomiasis diagnosis. Here we describe the optimisation of a novel droplet digital PCR (ddPCR) duplex assay for the diagnosis of Schistosoma japonicum infection which provides improved detection sensitivity and specificity. The assay involves the amplification of two specific and abundant target gene sequences in S. japonicum; a retrotransposon (SjR2) and a portion of a mitochondrial gene (nad1). The assay detected target sequences in different sources of schistosome DNA isolated from adult worms, schistosomules and eggs, and exhibits a high level of specificity, thereby representing an ideal tool for the detection of low levels of parasite DNA in different clinical samples including parasite cell free DNA in the host circulation and other bodily fluids. Moreover, being quantitative, the assay can be used to determine parasite infection intensity and, could provide an important tool for the detection of low intensity infections in low prevalence schistosomiasis-endemic areas.

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Objectives: Elevated shame and dissociation are common in dissociative identity disorder (DID) and chronic posttraumatic stress disorder (PTSD) and are part of the constellation of symptoms defined as complex PTSD. Previous work examined the relationship between shame, dissociation, and complex PTSD and whether they are associated with intimate relationship anxiety, relationship depression, and fear of relationships. This study investigated these variables in traumatized clinical samples and a nonclinical community group.

Method: Participants were drawn from the DID (n = 20), conflict-related chronic PTSD (n = 65), and nonclinical (n = 125) populations and completed questionnaires assessing the variables of interest. A model examining the direct impact of shame and dissociation on relationship functioning, and their indirect effect via complex PTSD symptoms, was tested through path analysis.

Results: The DID sample reported significantly higher dissociation, shame, complex PTSD symptom severity, relationship anxiety, relationship depression, and fear of relationships than the other two samples. Support was found for the proposed model, with shame directly affecting relationship anxiety and fear of relationships, and pathological dissociation directly affecting relationship anxiety and relationship depression. The indirect effect of shame and dissociation via complex PTSD symptom severity was evident on all relationship variables.

Conclusion: Shame and pathological dissociation are important for not only the effect they have on the development of other complex PTSD symptoms, but also their direct and indirect effects on distress associated with relationships.

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É incontornável o estatuto que o conceito de qualidade de vida assume hoje na prática e políticas de saúde pública. Na infância e adolescência é ainda escassa a investigação, tornando-se crucial o desenvolvimento de instrumentos de qualidade vida relacionada com a saúde validados para esta população. O presente trabalho tem como objetivo fundamental analisar as qualidades psicométricas e validar a versão portuguesa do Youth Quality of Life (YQOL-R) (Patrick, et al., 2002). A amostra é constituída por 507 adolescentes, com idades compreendidas entre os 12 e os 19 anos, a frequentar o 3º ciclo do ensino básio e ensino secundário de escolas públicas do ensino regular. Para além do citado instrumento a validar, os jovens preencheram também, para a análise da validade convergente e divergente, o Kidscreen-27 (Gaspar & Matos, 2008) e a Escala da Depressão, Ansiedade e Stresse (EADS-21) (Pais-Ribeiro, Honrado & Leal, 2004). Os resultados mostram que o YQOL-R apresenta uma estrutura fatorial de quatro fatores, semelhantes à versão original americana (individual, relações sociais, ambiente e qualidade de vida em geral). Possui uma boa consistência interna e uma adequada estabilidade temporal. Mostrou correlações significativas e no sentido esperado com as variáveis em estudo. Foram igualmente encontradas diferenças de género em relação à qualidade de vida, sendo os rapazes a reportarem em média níveis mais elevados de perceção da qualidade de vida, comparativamente às raparigas. Futuros estudos devem ser realizados em amostras clínicas para confirmação dos dados. Não obstante esta limitação, o presente estudo contribuiu para a disponibilização de um novo instrumento para avaliação da qualidade de vida em crianças e adolescentes, o qual evidenciou boas propriedade psicométricas, apoiando, assim empiricamente, a sua utilização nas práticas de saúde e investigação em amostras da comunidade. / Nowadays, it´s unavoidable the status that, the concept of quality of life assumes in practices and politics of public health. In childhood and adolescence it´s weak the investigation but it´s crucial the development of instruments of quality of life related to health validated to this population. The present work has an important aim, it´s analyze the psychometric qualities and validate the Portuguese version of Youth Quality of Life (YQOL-R) (Patrick, et al., 2002). The sample consists of 507 adolescents, aged between 12 and 19 years old and they attend the 3 rd cycle of basic education and secondary education schools in villages of regular education. Apart from that instrument, teenagers fill in, also, to the analysis convergent and divergent, the Kidscreen-27 (Gaspar & Matos, 2008) and the Depression, Anxiety and Stress Scale (EADS-21) (Pais-Ribeiro, Honrado & Leal, 2004). The results show that the YQOL-R presents a factorial structure with four factors similar to original American version (individual, social relations, environment and general quality of life), presenting a good internal consistence and an adequate temporal stability. Substantial correlations showed and in the expected way with the variables in study. Have been found differences of gender related to quality of life, boys reported on average higher levels of quality of life perception, comparatively to the girls. Future studies must be performed in clinical samples to confirm the findings. In spite of this restriction, the actual study contributes to providing a new instrument to evaluate the quality of life in children and adolescents, this evidenced good psychometric properties, supported empirically its application in heath practices and investigation in community samples.

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A presente investigação pretendeu validar para a população portuguesa de adolescentes a Escala de Memórias Precoces de Calor e Segurança em Relação ao Grupo de Pares (EMPCSPares) bem como ver cumpridos os seus principais objetivos: 1) Adaptação da escala de memórias precoces de calor e segurança (EMPCS), enquanto medida global, para o contexto de interação com o grupo de pares; 2) Explorar a validade de construto através da análise fatorial exploratória (estudo da dimensionalidade); 3) Analisar a consistência interna do instrumento e explorar a qualidade dos itens; 4) Examinar a validade convergente e divergente através da associação com outras variáveis semelhantes e distintas do construto em análise; 5) Analisar possíveis efeitos das variáveis sociodemográficas, como a idade, género e escolaridade nos resultados da escala, bem como a sua associação com a perceção global de qualidade vida; 6) Comparar as memórias precoces emocionais em função da qualidade de vinculação (segura e insegura). A amostra é constituída por 354 jovens (152 rapazes e 202 raparigas), com idades compreendidas entre os 12 e os 18 anos (M= 15,81; DP = 1,58) a frequentar o ensino básico e secundário do sistema regular de ensino público (7ºano de escolaridade ao 12º ano de escolaridade). Do protocolo constam os seguintes instrumentos: escala de memórias precoces de calor e segurança no contexto familiar (EMPCSFamília) e na interação com o grupo de pares (EMPCSPares); escala de auto-compaixão (SCS-A); escala de vergonha externa (OAS – A); questionário de vinculação (AQ-C); escala de ansiedade, depressão e stress (EADS-21). Os resultados obtidos mostram que a escala tem uma estrutura unidimensional, possui uma excelente consistência interna, uma estabilidade temporal adequada, assim como uma boa validade convergente e divergente. Revelou igualmente discriminar os jovens com uma vinculação segura dos que apresentam uma vinculação insegura. Apesar da necessidade de mais estudos, nomeadamente a realizar em amostras clínicas, a EMPCSPares mostrou ser um instrumento robusto e útil na avaliação de memórias emocionais no contexto de interação com os pares, constituindo um contributo relevante para a investigação e prática clínica com adolescentes. / This research intends to validate for the Portuguese population of adolescents the Early Memories Scale Heat and Safety Relative to Peer Group (EMPCSPares) and see fulfilled its main objectives: 1) The early warm and security memories scale adaptation (EMPCS), as a global measure for the interaction context with the peer group; 2) Explore the construct validity by exploratory factor analysis (study of dimensionality); 3) To analyze the internal consistency of the instrument and explore the quality of the items; 4) Examine the convergent and divergent validity by association with other similar and different variables of the construct in question; 5) Analyze possible effects of sociodemographic variables such as age, gender and education in the scale results, as well as its association with the global perception of quality of life; 6) compare the emotional early memories due to the link quality (secure and insecure). The sample consists of 354 children (152 boys and 202 girls), aged between 12 and 18 years (M = 15.81, SD = 1.58) attending basic and secondary education in the regular public education system (7th grade to 12th grade). The Protocol contains the following instruments: Scale of early heat and security memories in the family context (EMPCSFamily) and interaction with the peer group (EMPCSPairs) ; Self - compassion scale (SCS-A); external shame scale (OAS - A); linking questionnaire (AQ- C) ; scale of anxiety, stress and depression (EADS 21) . The results obtained show that the scale has a one-dimensional structure, has an excellent internal consistency, an adequate temporal stability as well as good convergent and divergent validity. It also showed discriminate young people with a secure attachment of those who have an insecure attachment. Despite the need for further studies, including the conduct of clinical samples, the EMPCSPares proved to be a robust and useful tool in the evaluation of emotional memories in the context of interaction with peers, constituting an important contribution to research and clinical practice with adolescents.

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RESUMO Objetivos: Uma nova versão portuguesa do teste Stroop é fundamental para a avaliação neuropsicológica. A versão portuguesa prévia incluía cores que muitas pessoas não conseguiam distinguir. Assim, é objetivo descrever as propriedades psicométricas de uma nova versão, designada versão Torga do Teste Stroop, numa amostra da população portuguesa. Métodos: Inserida no projeto Estudos Normativos de Instrumentos Neuropsicológicos, esta investigação conta com uma amostra global constituída por 544 participantes (241 homens e 303 mulheres) com idades compreendidas entre os 18 e os 97 anos. A avaliação foi realizada com recurso à versão Torga do Teste Stroop, à Figura Complexa de Rey-Osterrieth e à Bateria de Avaliação Frontal enquanto instrumentos de avaliação das funções executivas. Resultados: A versão Torga do Teste Stroop revelou uma consistência interna muito boa (α de Cronbach = 0,99). Revelou também adequada estabilidade temporal e validade convergente. Conclusão: A versão Torga do Teste Stroop aparenta ser um instrumento apropriado à avaliação neuropsicológica de adultos portugueses. Considerando a importância deste teste no contexto da avaliação neuropsicológica, incentivam-se estudos com novas amostras, incluindo amostras clínicas. ABSTRACT Goals: A new Portuguese version of the Stroop Test is essential for the neuropsychological assessment. The previous Portuguese version included colors that many people could not distinguish. Thus, it is aimed to describe the psychometric properties of a new version, called the Torga version of the Stroop Test in a sample of the Portuguese population. Methods: Being part of the Estudos Normativos de Instrumentos Neuropsicológicos/ Normative Studies of Neuropsychological Instruments, this research has a global sample of 544 subjects (241 men and 303 women) aged from 18 to 97 years. The assessment included the Torga version of the Stroop Test, the Rey-Osterrieth Complex Figure and the Frontal Battey Assessment, used as measures of executive functions. Results: The Torga version of the Stroop Test showed a very good internal consistency (Cronbach's α = 0.99). It also revealed an adequate temporal stability and convergent validity. Conclusion: The Torga version of the Stroop Test appears to be an adequate instrument for the neuropsychological assessment of Portuguese adults. Considering the importance of this test in the context of neuropsychological assessment, more studies with new samples, including clinical samples are encouraged.

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Stroke is currently one of the leading causes of death and disability worldwide. Despite recent advances in the treatment of stroke there is a major unmet clinical need for novel therapeutics for intervention. miRNAs are small coding RNAs which act to post-transcriptionally inhibit expression of genes. Emerging evidence has supported the view that miRNAs play an important role in the development and progression of ischaemic stroke, although understanding remains relatively poor. This research uses several models to investigate the effects of miRNAs in the context of stroke in vivo and in vitro, as well as assessment of patient serum samples in order to identify biomarkers for stroke. miR-29b was found to be significantly upregulated in SHRSP rat brain peri-infarct at 72h following stroke, and downregulated in ischaemic core at 24h and 72h following stroke, whilst miR-29c was significantly downregulated in remainder tissue at 24h following stroke and in infarct at 72h following stroke. The upreglation of miR-29b at 72h corresponded to a significant downregulation of miR-29 target genes MMP2, MMP9 and TGF-β1 in peri-infarct tissue at 72h following stroke. Modulation of miR-29b and miR-29c was achieved in a rat neuronal cell line but suppression of genes of interest was not observed following oxygen glucose deprivation. Several candidate miRNAs were then identified by microRNA Openarray analysis in stroke patient serum samples. Validation of these miRNAs was not demonstrated in the population studied, but assessment of these miRNAs in rat serum and isolated exosomes demonstrated that several of these miRNAs were significantly altered in SHRSP rats following stroke. Finally miR-21 was demonstrated to be significantly upregulated in SHRSP rat peri-infarct following stroke. This was associated with a change in miR-21 localization as determined by in situ hybridization. Modulation of miR-21 via the use of CAG-miR-21 mice demonstrated no difference in infarct size as measured by T2 -weighted MRI scan nor was any difference present in behavioural tests versus wild type. KO of miR-21 resulted in a reduction of survival rate compared with wild type. This thesis demonstrates that miR-29 and miR-21 are modulated following stroke in animal models, and these are potential candidates for therapeutic intervention in the future. Analysis of clinical samples has illustrated difficulties in the identification of serum miRNA profiles and suggests that looking at the exosomal component of serum may provide better information regarding miRNA profiles after stroke.

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Rotavirus double-stranded RNA was detected directly in sewage treatment plant samples over a 1-year period by reverse transcription followed by PCR amplification of the VP7 gene and Southern blot hybridization. The presence of naturally occurring rotaviruses was demonstrated in 42% of raw sewage samples and in 67% of treated effluent samples, Amplified viral sequences were analyzed bg restriction enzymes. Ten different restriction profiles were characterized, most of which were found in treated effluent samples. A mixture of restriction profiles was observed in 75% of contaminated effluent samples, The profiles were compared with those obtained from human rotavirus isolates involved in infections in children from the same area (six different profiles were detected), Five identical viral sequences were detected in both environmental and clinical samples, Restriction profiles sere also compared io profiles from known genomic sequences of human and animal viruses. Both human and animal origins of rotavirus contamination of water seemed likely.

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Parenting is a robust predictor of developmental outcomes among children with ADHD. Early parenting predicts the persistence and course of ADHD and comorbid problems above and beyond risk associated with shared genetic effects. Yet, on average, mothers of children with ADHD are less positive and more negative in their parent-child interactions compared to mothers of non-disordered children. Little is known about psychobiological markers which may be associated with individual variations in maternal parenting in families of children with ADHD. Neurobiological models of parenting suggest that maternal cortisol levels following a stressor may be positively associated with hostile and intrusive parenting; however, to date no studies have examined maternal cortisol reactivity and parenting in school-age, or clinical samples of, children. Mothers’ regulation of physiological stress responses may be particularly important for families of children with ADHD, as parenting a child with chronically challenging behaviors represents a persistent environmental stressor. The current study sought to extend the existing literature by providing an empirical examination of the relationship between maternal cortisol reactivity following two laboratory stressors and parenting among mothers of children with and without ADHD. It was hypothesized that child ADHD group would moderate the relationship between cortisol reactivity and self-reported and observed parenting. Greater total cortisol output and greater increase in cortisol during the TSST were associated with decreased positive parenting and increased negative and directive parenting, with the exception of parental involvement, which was associated with increased cortisol output during the TSST. Conversely, cortisol output during the PCI was associated with increased positive parenting, increased parental involvement, and decreased negative parenting. In contrast to the TSST, a greater decrease in cortisol during the PCI indicated more positive parenting and parental involvement. These associations were specific to mothers of children with ADHD, with the exception of maternal directiveness, which was specific to comparison mothers. Findings add to our understanding of physiological processes associated with maternal parenting and contribute to an integrative biological, psychological, and cognitive process model of parenting in families of children with ADHD.