FoxP3 T cells and the pathophysiologic effects of brain death and warm ischemia in donor kidneys

Background and objectives Forkhead box P3 regulatory T cells control inflammatory responses, but it remains unclear whether they inhibit brain death-initiated inflammation and tissue injury in deceased kidney donors. Design, setting, participants, & measurement To study the actions of regulatory T cells at various stages of the donation and transplantation procedure, forkhead box P3, regulatory and inflammatory cytokine expression, and tissue injury markers were determined in time 0 kidney biopsies from deceased and living donors. Additionally, the interaction between forkhead box P3+ T cells and kidney injury molecule-1 by activated primary tubular epithelial cells was studied. Results After cold storage, the deceased donor kidneys expressed the higher mRNA levels of kidney injury molecule-1 and CD3ε. In these samples, the inflammatory cytokines IL-8 and IFN-γ and markers associated with regulation (forkhead box P3, TGF-β, and IL-10) were highly expressed compared with living donor kidneys. Correlations were found between mRNA expression levels of forkhead box P3 and kidney injury molecule-1 and forkhead box P3 and IFN-γ. Immunohistochemical analysis confirmed the presence of forkhead box P3+ T cells in donor kidneys. Renal function (analyzed by serum creatinine levels) at the first week posttransplantation correlated with kidney injury molecule-1 and forkhead box P3 mRNA levels. In vitro studies showed that kidney injury molecule-1 expression by primary tubular epithelial cells was 63% (mean) lower when cocultured with regulatory T cells compared with control T cells. Conclusions These results show that donor forkhead box P3+ T cells infiltrate the deceased donor kidney, where they may control inflammatory and injury responses

Genetic analysis of IRF6, a gene involved in craniofacial midline formation, in relation to pituitary and facial morphology of patients with idiopathic growth hormone deficiency

textabstractIntroduction: Growth hormone is secreted by the pituitary gland, which forms part of the craniofacial midline. IRF6 encodes a transcription factor involved in the development of the craniofacial midline and mutations in IRF6 are known to disturb craniofacial development. Craniofacial and pituitary development are closely related. After whole exome sequencing revealed a new mutation in IRF6 in a family with Idiopathic Growth Hormone Deficiency (IGHD), we screened the remainder of our IGHD cohort for mutations in this gene and related their genotypes to pituitary and craniofacial morphology. Materials and methods: We sequenced all coding exons and exon–intron boundaries of IRF6 in 81 patients with IGHD. We performed a multiplex ligation-dependent probe amplification (MLPA) in order to exclude copy number variations in IRF6. We analyzed facial measurements taken from standardized digital pictures of 48 patients. Results: We found two new variants and eleven polymorphisms. Apart from the new mutation found in the index family (p.Arg233Cys), we found one other new heterozygous missense mutation in IRF6 (Pro456Ser). p.Arg233Cys was reported as extremely rare in exome databases (1 allele out of 120.852 alleles sequenced), strictly conserved among species and was predicted deleterious by all variant predictor programs. Pro456Ser was predicted to be benign. MLPA did not reveal any exon deletions or duplications in any of the patients. Conclusion: This is the first report of IRF6 analysis in an IGHD cohort. We found one new mutation which, based on in silico analysis, could be of functional relevance. However, we did not find any mutations in the other patients. Therefore, we conclude that IRF6 defects are rare in IGHD patients and further research should focus on new candidate genes

Measuring chronic care management experience of patients with diabetes: PACIC and PACIC+ validation

Background: The patient assessment of chronic illness care (PACIC) is a promising instrument to evaluate the chronic care experiences of patients, yet additional validation is needed to improve its usefulness.Methods: A total of 1941 patients with diabetes completed the questionnaire. Reliability coefficients and factor analyses were used to psychometrically test the PACIC and PACIC+ (i.e., PACIC extended with six additional multidisciplinary team functioning items to improve content validity). Intra-Class Correlations were computed to identify the extent to which variation in scores can be attributed to GP practices. Results: The PACIC and PACIC+ showed a good psychometric quality (Cronbach's alpha's > 0.9). Explorative factor analyses showed inconclusive results. Confirmative factor analysis showed that none of the factor structures had an acceptable fit (RMSEA > 0.10). In addition, 5.1 to 5.4 percent of the total variation was identified at the GP practice level. Conclusion: The PACIC and PACIC+ are reliable instruments to measure the chronic care management experiences of patients. The PACIC+ is preferred because it also includes multidisciplinary coordination and cooperation - one of the central pillars of chronic care management - with good psychometric quality. Previously identified subscales should be used with caution. Both PACIC instruments are useful in identifying GP practice variation

Thymus-Derived Regulatory T Cells Infiltrate the Cardiac Allograft Before Rejection

Background. FOXP3+ regulatory T cells (Treg) either originate in the thymus (natural [n]Treg) or are induced in the periphery by antigen exposure and cytokines (induced [i]Treg). It is currently not elucidated which and to what extent these Treg subsets regulate intracardiac allogeneic responses in transplant patients. Methods. By using demethylation of the Treg-specific demethylated region in the FOXP3 gene as a marker for nTreg and FOXP3 messenger RNA expression as a marker for the total Treg population, we examined Treg in endomyocardial biopsies (EMBs) of both patients who developed an acute rejection necessitating therapy (rejectors; International Society for Heart and Lung Transplantation rejection grade >= 2R) and patients who remained free from rejection (nonrejectors). Results. In the presence of comparable messenger RNA levels of CD3, IL-10, TGF beta, IL2, IFN gamma, and IL17A, the percentage of nTreg was significantly higher in EMB with histological signs of mild rejection (rejection grade 1R) collected before rejection than in 1R EMB of nonrejectors. The total Treg population was comparable in 1R EMB of nonrejectors and 1R EMB collected before rejection, which suggests the presence of iTreg in the EMB of nonrejectors. The relative high percentage of nTreg after rejection was not related to the number of rejections, whereas the total Treg population was inversely related to the number of rejections the first year after transplantation. Conclusions. Our data indicate that intragraft nTreg are unable to restrain alloreactivity leading to rejection. Moreover, the indirect evidence of the presence of intragraft iTreg suggests a possible role of iTreg in the regulation of alloreactivity

Quality of diabetes care in Dutch care groups: no differences between diabetes patients with and without co-morbidity

<strong>Objective</strong>: To evaluate the relationship between presence and nature of co-morbidity and quality of care for diabetes patients enrolled in diabetes disease management programmes provided by care groups. <strong>Methods</strong>: We performed an observational study within eight Dutch diabetes care groups. Data from patient record systems of care groups and patient questionnaires were used to determine quality of care. Quality of care was measured as provision of the recommended diabetes care, patients’ achievement of recommended clinical outcomes and patients’ perception of coordination and integration of care. <strong>Results</strong>: 527 diabetes patients without and 1187 diabetes patients with co-morbidity were included. Of the co-morbid patients, 7.8% had concordant co-morbid conditions only, 63.8% had discordant co-morbid diseases only and 28.4% had both types of conditions. Hardly any differences were observed between patients with and without co-morbidity in terms of provided care, achievement of clinical outcomes and perceived coordination and integration of care. <strong>Conclusions</strong>: Our study implies that care groups are able to provide similar quality of diabetes care for diabetes patients with and without co-morbidity. Considering the expected developments regarding additional disease management programmes in care groups, it is of importance to monitor quality of care, including patient experiences, for all chronic diseases. It will then become clear whether accountable provider-led organisations such as care groups are able to ensure quality of care for the increasing number of patients with multiple chronic conditions