Intra- and inter-day variation of cytokines and chemokines in tears of healthy subjects.

Tear levels of certain cytokines/chemokines can potentially serve as biomarkers for dry eye and other ocular surface diseases if they remain stable from day-to-day in healthy eyes. The aim of this study was to determine the normal intra- and inter-day variation of selected tear cytokines/chemokines. Tear samples from 24 young, healthy adults were collected 11:00 AM-1:00 PM (mid-day) and 5:00-7:00 PM (evening) on three non-consecutive days. Concentrations of 18 cytokines/chemokines (EGF, eotaxin, CX3CL1/fractalkine, GM-CSF, IFN-γ, IL-10, IL-1ß, IL-13, IL-17A, IL-1RA, IL-5, IL-6, CXCL8/IL-8, IL-9, CXCL10/IP-10, CCL5/RANTES, TNF-α, and VEGF) were measured by multiplex bead analysis. Ocular surface disease was ruled out by clinical tests. A random-effects ANOVA model was used to evaluate intra- and inter-day effects on cytokine/chemokine levels. Repeatability of intra-subject inter-day measurements was assayed by coefficient of variation. Ten out of the 18 molecules had detectable tear levels in >50% of the subjects. Of those, only IL-10 and IL-1ß levels had significant inter-day variations. EGF, CX3CL1/fractalkine, CXCL10/IP-10, and VEGF were consistently higher in the evening compared to the mid-day measurements. EGF, CXCL10/IP-10, VEGF and CXCL8/IL-8had good intra-subject reproducibility. In conclusion, tear cytokines/chemokines can be measured reproducibly over time, with most not having significant inter-day variability. Some varied significantly depending upon the time of tear collection, and these variations should be taken into account when comparisons are made. The good intra-subject reproducibility for EGF, CXCL10/IP-10, CXCL8/IL-8, and VEGF indicates that these molecules could potentially serve as biomarkers of ocular surface disease.

Influence of a controlled environment simulating an in-flight airplane cabin on dry eye disease.

PURPOSE: To evaluate symptoms, signs, and the levels of 16 tears inflammatory mediators of dry eye (DE) patients exposed to an environment simulating an in-flight air cabin in an environmental chamber. METHODS: Twenty DE patients were exposed to controlled environment simulating an in-flight airplane cabin (simulated in-flight condition [SIC]) of 23°C, 5% relative humidity, localized air flow, and 750 millibars (mb) of barometric pressure. As controls, 15 DE patients were subjected to a simulated standard condition (SSC) of 23°C, 45% relative humidity, and 930 mb. A DE symptoms questionnaire, diagnostic tests, and determination of 16 tear molecules by multiplex bead array were performed before and 2 hours after exposure. RESULTS: After SIC exposure, DE patients became more symptomatic, suffered a significant (P ≤ 0.05) decrease in tear stability (tear break up time) (from 2.18 ± 0.28 to 1.53 ± 0.20), and tear volume (phenol red thread test), and a significant (P ≤ 0.05) increase in corneal staining, both globally (0.50 ± 0.14 before and 1.25 ± 0.19 after) and in each area (Baylor scale). After SSC, DE patients only showed a mild, but significant (P ≤ 0.05), increase in central and inferior corneal staining. Consistently, tear levels of IL-6 and matrix metalloproteinase (MMP)-9 significantly increased and tear epidermal growth factor (EGF) significantly decreased (P ≤ 0.05) only after SIC. CONCLUSIONS: The controlled adverse environment conditions in this environmental chamber can simulate the conditions in which DE patients might be exposed during flight. As this clearly impaired their lacrimal functional unit, it would be advisable that DE patients use therapeutic strategies capable of ameliorating these adverse episodes.

Effect of TGF-ß on ocular surface epithelial cells.

A role for transforming growth factor (TGF)-ß in the pathogenesis of some ocular surface diseases has been proposed. We determined if secretion of TGF-ß and expression of TGF-ß receptors RI, RII, and RIII by human ocular surface epithelial cells were modified under inflammatory conditions. We also determined how these cells responded to TGF-ß. A human corneal epithelial (HCE) cell line and a conjunctival epithelial cell line (IOBA-NHC) were exposed to TGF-ß1 and -ß2 and to proinflammatory cytokines. TGF-ß receptor mRNAs were analyzed by real time reverse transcription polymerase chain reaction (RT-PCR) in both cell lines, and in conjunctival, limbal, and corneal epithelial cells from post-mortem human specimens. Expression of TGF-ß receptors and pSMAD2/SMAD2 were determined by Western blot and immunofluorescence assays. Secretion of TGF-ß isoforms, cytokine/chemokine, and metalloproteinases (MMPs) were analyzed in cell supernatants by immunobead-based assays. Secretory leukocyte proteinase inhibitor (SLPI) secretion was analyzed by enzyme-linked immunosorbent assay. TGF-ß isoform and receptor gene expression was determined by RT-PCR in conjunctival epithelium of dry eye (DE) patients and healthy subjects. Our results showed that TGF-ß RI expression was down-regulated with IL-4 exposure, whereas TGF-ß RII and TGF-ß2 were upregulated by TNF-α in HCE cells. TGF-ß RIII receptor expression was upregulated in IOBA-NHC cells by TNF-α and IFN-γ. SMAD2 phosphorylation occurred in HCE and IOBA-NHC cells after TGF-ß treatment. TGF-ß significantly up- and down-regulated secretion of several cytokines/chemokines by both cell lines and MMP by HCE cells. TGF-ß2 and TGF-ß3 were upregulated and TGF-ß RIII mRNA was down-regulated in DE conjunctival epithelium. These results show that TGF-ß plays an important role in directing local inflammatory responses in ocular surface epithelial cells.

Gasto farmacéutico de médicos de atención primaria del área de salud de Cuenca / Pharmaceutical expenditure among primary care physicians in the province of Cuenca (Spain)

Objetivos El trabajo pretende determinar los elementos clave que definen el gasto farmacéutico por médico del Área de Salud de Cuenca (España). Conocerlos puede ser relevante para la política sanitaria, al tener información que permita tomar decisiones para incidir en la contención del gasto. También podría ser de interés para médicos de atención primaria, especialistas y autoridades sanitarias de Cuenca y de otras áreas de salud, y para investigadores en gestión sanitaria y economía de la salud. Métodos Se desarrolla un análisis de regresión lineal múltiple para tres variables dependientes con el fin de conocer qué características permiten explicar la variabilidad en la prescripción de genéricos, la tasa de derivación y el gasto farmacéutico de los médicos de atención primaria.Resultados No ser médico interno residente (MIR), ejercer en un municipio de más de 10.000 habitantes y con un cupo de gran tamaño, supuso un impacto negativo en el porcentaje de medicamentos genéricos prescritos. Los médicos que usan la historia clínica informatizada y derivan más a especializada, prescriben menos genéricos. Las dos variables más importantes para explicar el elevado gasto farmacéutico son el mayor tamaño del cupo y el alto porcentaje de pacientes mayores de 65 años. El mayor número de genéricos reducía el gasto, y el incremento en el porcentaje de pacientes mayores en el cupo lo aumentaba.Conclusiones Una de las variables que mejor explica el gasto farmacéutico es el porcentaje de genéricos prescritos, teniendo en cuenta que se controla por variables tan importantes como tamaño del cupo o porcentaje de pacientes de edad avanzada incluidos en él (AU)

Objective To determine the factors related to pharmaceutical expenditure per primary care physician in the province of Cuenca, which could allow public health authorities to take decisions to curb expenditure. Determining these factors could also be of interest to primary care physicians, specialists, health authorities, health economists, and health management researchers.Methods We performed a multiple linear regression analysis with three dependent variables to determine which factors explain variability in generic drug prescription, the referral rate, and pharmaceutical expenditure among primary care physicians.Results Not being a resident physician, practicing in a town with more than 10,000 people and having a large patient list had a negative impact on the percentage of generic drugs prescribed. Physicians using electronic medical records and with a high referral rate to specialized care prescribed fewer generic drugs. The two most important factors explaining pharmaceutical expenditure were having a large patient list and a high percentage of patients aged more than 65 years old. Expenditure was reduced by greater prescription of generic drugs and was increased by a larger proportion of elderly patients on the patient list.Conclusions After controlling for the size of the patient list, the proportion of elderly patients and other factors, the percentage of generic drugs prescribed was one of the most important factors explaining pharmaceutical expenditure in primary care physicians (AU)

[Pharmaceutical expenditure among primary care physicians in the province of Cuenca (Spain)]. / Gasto farmacéutico de médicos de atención primaria del área de salud de Cuenca.

OBJECTIVE: To determine the factors related to pharmaceutical expenditure per primary care physician in the province of Cuenca, which could allow public health authorities to take decisions to curb expenditure. Determining these factors could also be of interest to primary care physicians, specialists, health authorities, health economists, and health management researchers. METHODS: We performed a multiple linear regression analysis with three dependent variables to determine which factors explain variability in generic drug prescription, the referral rate, and pharmaceutical expenditure among primary care physicians. RESULTS: Not being a resident physician, practicing in a town with more than 10,000 people and having a large patient list had a negative impact on the percentage of generic drugs prescribed. Physicians using electronic medical records and with a high referral rate to specialized care prescribed fewer generic drugs. The two most important factors explaining pharmaceutical expenditure were having a large patient list and a high percentage of patients aged more than 65 years old. Expenditure was reduced by greater prescription of generic drugs and was increased by a larger proportion of elderly patients on the patient list. CONCLUSIONS: After controlling for the size of the patient list, the proportion of elderly patients and other factors, the percentage of generic drugs prescribed was one of the most important factors explaining pharmaceutical expenditure in primary care physicians.

Calcineurin inhibitors, but not rapamycin, reduce percentages of CD4+CD25+FOXP3+ regulatory T cells in renal transplant recipients.

BACKGROUND: Immunosuppression in renal transplantation, although manageable in the short-term, is a major hurdle for long-term graft survival. Recently, increased frequencies of CD4CD25 regulatory T cells (Tregs) have been described as an additional mechanism that induces alloimmune tolerance. METHODS: We assessed 64 renal transplant recipients with stable renal function for at least one year. Patients were divided into two groups according to the immunosuppression they were receiving at the moment of the study: one consisted of patients receiving rapamycin (Rapa) but not calcineurin inhibitors (CNI), and the other group received CNI but not Rapa. The Rapa group was further divided into three subgroups according to their previous experience with CNI: CNI-free, CNI withdrawal, and CNI conversion. Frequencies of blood Tregs were studied by flow cytometry after staining with monoclonal antibodies specific for different markers of Tregs. RESULTS: Frequencies of CD4 T cells with regulatory phenotype and function were significantly decreased in peripheral blood of renal transplant patients receiving CNI compared with those receiving Rapa. This effect was independent of an early exposure to CNI because the CNI-free patients in the Rapa group showed similar frequencies of Tregs to the CNI withdrawal and CNI conversion groups. CONCLUSIONS: CNI, but not Rapa, induce a decrease of circulating Tregs in stable renal transplant recipients. Thus, Rapa might be further explored in strategies using preservation of Tregs for transplant tolerance. Furthermore, quantification of blood Tregs may be a suitable tool to identify renal transplant recipients who may be candidates for reduced immunosuppression.