Pharmacogenetics of trough serum anti-TNF levels in paediatric inflammatory bowel disease.

AIMS: Identifying DNA variants associated with trough serum anti-tumour necrosis factor (TNF) levels could predict response to treatment in inflammatory bowel disease (IBD). To date, no specific studies have been performed in children. The aim of this study was to identify genetic variants associated with trough serum anti-TNF levels and whether these variants are differential markers for infliximab and adalimumab. METHODS: We included 154 children (age < 18 years) from 17 hospitals who had been diagnosed with IBD and actively treated with infliximab or adalimumab. Twenty-one polymorphisms were genotyped using real-time PCR. Trough serum anti-TNF levels were measured using enzyme-linked immunosorbent assay (ELISA). The association between DNA polymorphisms and the therapeutic range or the absolute values of anti-TNF drugs was analysed by Fisher exact test, student's t-test and logistic regression. RESULTS: The variants rs5030728 (TLR4) and rs11465996 (LY96) were associated with subtherapeutic infliximab levels. rs1816702 (TLR2) was associated with supratherapeutic levels and rs3397 (TNFRSF1B) with subtherapeutic levels of adalimumab (P < .05). In addition, rs1816702 (TLR2) and rs2569190 (CD14) were associated with absolute values of trough serum adalimumab, and rs2569190 (CD14) was associated with absolute values of trough serum adalimumab and infliximab (P < .05). CONCLUSION: Genotyping of these DNA variants before starting treatment may help to select the best anti-TNF drug in paediatric patients. The SNP rs1816702 is the most promising marker for tailoring the anti-TNF regimen in children with IBD. For the first time, DNA variants are associated with trough serum anti-TNF levels.

Genotyping of UGT1A1*80 as an Alternative to UGT1A1*28 Genotyping in Spain.

BACKGROUND: The variant rs34983651 (UGT1A1*28) and its genotyping are used to prevent irinotecan-induced toxicity. Several variants are in close linkage disequilibrium. Our objective was to evaluate the potential correlation of genotyping UGT1A1*80 instead of UGT1A1*28 in different populations. METHODS: We studied SNPs in linkage disequilibrium with UGT1A1*28 in several populations and selected rs887829 to develop an inexpensive and rapid genotyping method and compare it with the one we currently use for UGT1A1*28 genotyping. Samples from cancer patients (n = 701) already tested using PCR and electrophoresis prior to treatment with irinotecan for rs34983651 (UGT1A1*28) in a Spanish hospital were genotyped for rs887829 (UGT1A1*80) using real-time PCR with a TaqMan probe. RESULTS: We observed a complete match for both genotypes, except in one sample. This method was 100% efficient in correctly genotyping *28/*28 patients, 99.68% efficient for *1/*28, and 100% efficient for *1/*1. Linkage disequilibrium between populations showed the Iberian population to be the most suitable for the clinical use of UGT1A1*80. This method is less expensive and the time to decision is shorter. CONCLUSION: Genotyping of rs887829 using the proposed method may be used to substitute genotyping of rs34983651 as a pharmacogenetics test in cancer patients prior to starting irinotecan-based treatments, mainly in the Iberian population. In addition, it is less expensive than other conventional methods and easy to implement, with a shorter time to decision than UGT1A1*28.

ABO blood group is involved in the quality of the specific immune response anti-SARS-CoV-2.

Since December 2019, the coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread throughout the world. To eradicate it, it is crucial to acquire a strong and long-lasting anti-SARS-CoV-2 immunity, by either natural infection or vaccination. We collected blood samples 12-305 days after positive polymerase chain reactions (PCRs) from 35 recovered individuals infected by SARS-CoV-2. Peripheral blood mononuclear cells were stimulated with SARS-CoV-2-derived peptide pools, such as the spike (S), nucleocapsid (N) and membrane (M) proteins, and we quantified anti-S immunoglobulins in plasma. After 10 months post-infection, we observed a sustained SARS-CoV-2-specific CD4+ T-cell response directed against M-protein, but responses against S- or N-proteins were lost over time. Besides, we demonstrated that O-group individuals presented significantly lower frequencies of specific CD4+ T-cell responses against Pep-M than non O-group individuals. The non O-group subjects also needed longer to clear the virus, and they lost cellular immune responses over time, compared to the O-group individuals, who showed a persistent specific immune response against SARS-CoV-2. Therefore, the S-specific immune response was lost over time, and individual factors might determine the sustainability of the body's defenses, which must be considered in the future design of vaccines to achieve continuous anti-SARS-CoV-2 immunity.

Gender and the professional career of primary care physicians in Andalusia (Spain).

BACKGROUND: Although the proportion of women in medicine is growing, female physicians continue to be disadvantaged in professional activities. The purpose of the study was to determine and compare the professional activities of female and male primary care physicians in Andalusia and to assess the effect of the health center on the performance of these activities. METHODS: Descriptive, cross-sectional, and multicenter study. SETTING: Spain. POPULATION: urban health centers and their physicians. SAMPLE: 88 health centers and 500 physicians. INDEPENDENT VARIABLE: gender. MEASUREMENTS: Control variables: age, postgraduate family medicine specialty (FMS), patient quota, patients/day, hours/day housework from Monday to Friday, idem weekend, people at home with special care, and family situation. DEPENDENT VARIABLES: 24 professional activities in management, teaching, research, and the scientific community. Self-administered questionnaire. Descriptive, bivariate, and multilevel logistic regression analyses. RESPONSE: 73.6%. Female physicians: 50.8%. Age: female physicians, 49.1 ± 4.3 yrs; male physicians, 51.3 ± 4.9 yrs (p < 0.001). Female physicians with FMS: 44.2%, male physicians with FMS: 33.3% (p < 0.001). Female physicians dedicated more hours to housework and more frequently lived alone versus male physicians. There were no differences in healthcare variables. Thirteen of the studied activities were less frequently performed by female physicians, indicating their lesser visibility in the production and diffusion of scientific knowledge. Performance of the majority of professional activities was independent of the health center in which the physician worked. CONCLUSIONS: There are gender inequities in the development of professional activities in urban health centers in Andalusia, even after controlling for family responsibilities, work load, and the effect of the health center, which was important in only a few of the activities under study.

Comprehensive Flow Cytometry Profiling of the Immune System in COVID-19 Convalescent Individuals.

SARS-CoV-2 has infected more than 200 million people worldwide, with more than 4 million associated deaths. Although more than 80% of infected people develop asymptomatic or mild COVID-19, SARS-CoV-2 can induce a profound dysregulation of the immune system. Therefore, it is important to investigate whether clinically recovered individuals present immune sequelae. The potential presence of a long-term dysregulation of the immune system could constitute a risk factor for re-infection and the development of other pathologies. Here, we performed a deep analysis of the immune system in 35 COVID-19 recovered individuals previously infected with SARS-CoV-2 compared to 16 healthy donors, by flow cytometry. Samples from COVID-19 individuals were analysed from 12 days to 305 days post-infection. We observed that, 10 months post-infection, recovered COVID-19 patients presented alterations in the values of some T-cell, B-cell, and innate cell subsets compared to healthy controls. Moreover, we found in recovered COVID-19 individuals increased levels of circulating follicular helper type 1 (cTfh1), plasmablast/plasma cells, and follicular dendritic cells (foDC), which could indicate that the Tfh-B-foDC axis might be functional to produce specific immunoglobulins 10 months post-infection. The presence of this axis and the immune system alterations could constitute prognosis markers and could play an important role in potential re-infection or the presence of long-term symptoms in some individuals.

DPYD Exome, mRNA Expression and Uracil Levels in Early Severe Toxicity to Fluoropyrimidines: An Extreme Phenotype Approach.

Dihydropyrimidine dehydrogenase deficiency is a major cause of severe fluoropyrimidine-induced toxicity and could lead to interruption of chemotherapy or life-threatening adverse reactions. This study aimed to characterize the DPYD exon sequence, mRNA expression and in vivo DPD activity by plasma uracil concentration. It was carried out in two groups of patients with extreme phenotypes (toxicity versus control) newly treated with a fluoropyrimidine, during the first three cycles of treatment. A novel nonsense gene variant (c.2197insA) was most likely responsible for fluoropyrimidine-induced toxicity in one patient, while neither DPYD mRNA expression nor plasma uracil concentration was globally associated with early toxicity. Our present work may help improve pharmacogenetic testing to avoid severe and undesirable adverse reactions to fluoropyrimidine treatment and it also supports the idea of looking beyond DPYD.

[The perspective of migrants on access to health care in the context of austerity policies in Andalusia (Spain)]. / La perspectiva de personas migrantes sobre el acceso a la atención sanitaria en el contexto de políticas de austeridad en Andalucía.

OBJECTIVE: To conduct an assessment of migrant people regarding their access to the health system following entry into force of Royal Decree-Law 16/2012 along with the impact of economic cuts on such access. METHOD: Qualitative phenomenological study with semi-structured interviews, conducted in Andalusia (Spain), in two phases (2009-2010 and 2012-2013), with 36 participants. The sample was segmented by length of stay, nationality and area of residence. The nationalities of origin are Bolivia, Morocco and Romania. RESULTS: Elements facilitating access in both periods: regular administrative situation, possession of Individual Health Card, knowledge of the language, social networks and information. The results show differences in access to health care for migrants before and after the enforcement of the RDL 16/2012, within austerity policies. In the second period, access barriers such as waiting times or incompatibility of schedules are aggravated and the socio-economic and administrative conditions of participants worsen. CONCLUSIONS: The design of policies, economic and regulatory health care, should take into account barriers and facilitators of access as fundamental main points of health protection for migrants and, therefore, for the general population.

[Characteristics of the family of origin and family formed by women and men primary care physicians of Andalusia, Spain]. / Características de la familia de origen y de la familia formada por las médicas y los médicos de familia de Andalucía.

BACKGROUND: The origin and formed family characteristics are related to physician's professional career. The purpose of this study was to know and compare by sex the characteristics of the origin family and formed family of women and men family physician in Andalusia. METHODS: Cross sectional and multicenter study. SETTING: Urban primary health care centres from Andalusian province capitals. PARTICIPANTS: Physician of primary health care centres. INCLUSION CRITERIA: at least one year using computerized medical history with the same quota patients. Multistage random sample, 88 primary health care centres and 500 physicians, 50% of both sexes (alpha=5%, power=90%, precision=15%). Postal auto administrated questionnaire. VARIABLES: sex, age, tutor of resident in family medicine, last father's activity, last mother's activity, number of brothers or sisters, family situation, last couple's activity (if any), to have or not children. RESULTS: 73.6 % responses. In no responses there weren't differences of sex neither tutor of resident in family medicine. Mean age: women physicians 49.5 +/- 4.3 and men physician 51.3 +/- 4.9 (p= or < 0.005). Postgraduate formation in family medicine: 42.2% of women and 33.3% of men (p=0.016). Live alone: 6.1% of women physician and 2.7% men physician (p=0.005). Live alone with children 9.9% of women and 2.2% of men (p=0.005). 16.5% of women and 10.2% of men don t have children (p=0.077). 21.1% of men physician s couples work only at home vs. 0.1% of women physician s couples (p= or < 0.005). 46% of women physician s couples is also physician vs. 22% of men physician s couples (p= or < 0.005). No significant differences registered in parent s activities neither in the number of brother or sisters. CONCLUSION: There are no significant differences in physician s origin family. However important differences in the characteristics of formed family are observed in both sexes.

Severe toxicity to capecitabine due to a new variant at a donor splicing site in the dihydropyrimidine dehydrogenase (DPYD) gene.

Severe, life-threatening adverse reactions to capecitabine sometimes occur in the treatment of solid tumors. Screening for dihydropyrimidine dehydrogenase (DPYD) deficiency is encouraged before start of treatment, but the genetic variants that are commonly analyzed often fail to explain toxicities seen in clinical practice. Here we describe the case of a 79-year-old Caucasian female with breast cancer who presented with life-threatening, rapidly increasing toxicity after 1 week of treatment with capecitabine and for whom routine genetic DPYD test resulted negative. DPYD exon sequencing found variant c.2242+1G>T at the donor splicing site of exon 19. This variant is responsible for skipping of exon 19 and subsequent generation of a non-functional DPYD enzyme. This variant has not been described previously but was found in three other members of the patient's family. With this case, we show that exon sequencing of DPYD in patients who experience marked toxicity to fluoropyrimidines and test negative for commonly evaluated variants can prove extremely useful for identifying new genetic variants and better explain adverse reactions causality.

Possible implication of ciliary neurotrophic factor (CNTF) and beta-synuclein in the ammonia effect on cultured rat astroglial cells: a study using DNA and protein microarrays.

Astrocytes are considered the key cell in hepatic encephalopathy; although their precise role in the disease has not yet been determined, exposure to ammonia appears to have an important pathogenic effect. We exposed confluent cultures of rat astroglial cells to ammonia (5mM NH(4)Cl) for 1, 3, 5 and 7 days, and determined astroglial levels of actin, glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), GLAST glutamate transporter, 25kDa heat-shock protein (HSP25), HSP60 and HSP70 by Western blot; the glutamine content in culture medium was measured by mass spectrometry. Significant increases were observed for GS, HSP60 and glutamine, and significant reductions for actin and GFAP. Astrocytes exposed to ammonia for 4 days were used to analyze the effect of ammonia in protein and DNA microarrays. After protein microarray data filtration by signal intensity, x-fold change and z-score, 11 proteins were selected, among which the significant increase in beta-synuclein was confirmed by Western blot. DNA microarray data filtration by intensity signal, x-fold change and p-value selected almost 600 genes. The significant increase in alpha-synuclein mRNA was confirmed by quantitative RT-PCR, but no change was observed in alpha-synuclein protein levels. A notable decrease in ciliary neurotrophic factor (CNTF) was demonstrated by Western blot after ammonia treatment, concurring with the reduction in CNTF mRNA observed in DNA microarrays. We discuss the possibility of a pathogenic role for CNTF and a protective role for beta-synuclein in experimental hyperammonemia. This study demonstrates the use of microarrays as tools to ascertain the possible implication of previously unidentified proteins in the pathogenesis of hepatic encephalopathy.

Gender inequalities in COPD decision-making in primary care.

BACKGROUND: COPD is a frequent severe illness that increasingly affects females. Gender inequalities have been reported in COPD care. OBJECTIVE: To analyze decision-making in primary care for men and women with identical COPD as a function of the gender of the family physician (FP). METHODS: Cross-sectional, multicenter study in 457 Andalusian FPs, using a self-administered vignette-based questionnaire on COPD featuring a male or female patient, with four variables on clinical reasoning: "tobacco as most important risk factor (RF)", "ordering of spirometry", "COPD as most likely diagnosis", and "referral". Multilevel logistic regression analysis. RESULTS: Response rate was 67.4% (308/457). In analysis of the four FP gender-patient gender dyads, tobacco was more frequently considered as priority RF for the man than for the woman in the vignette by female (95.6%vs.67.1%) and male (79.8%vs.62.5%) FPs. COPD was more frequently the most likely diagnosis for the man versus woman by female (84.4%vs.49.9%) and male (78.5%vs.57.8%) FPs. Male FPs more frequently ordered spirometry for the man versus woman (68.1%vs.46.8%). There were no differences in referral between male and female patients. Male FPs were more likely than female FPs to consider tobacco as priority RF for the man (p = 002). Female FPs were more likely than male FPs to refer the man (22.5%vs.8%). CONCLUSIONS: There may be gender inequalities in primary care for COPD in our setting. Diagnostic and therapeutic efforts appear lower in female patients. Male and female FPs only differed in care of the male patient, indicating FP gender-patient gender interaction.

Professional Success and Gender in Family Medicine: Design of Scales and Examination of Gender Differences in Subjective and Objective Success Among Family Physicians.

Two components of professional success have been defined: objective career success (OCS) and subjective career success (SCS). Despite the increasing number of women practicing medicine, gender inequalities persist. The objectives of this descriptive, cross-sectional, and multicenter study were (a) to construct and validate OCS and SCS scales, (b) to determine the relationships between OCS and SCS and between each scale and professional/family characteristics, and (c) to compare these associations between male and female family physicians (FPs). The study sample comprised 250 female and 250 male FPs from urban health centers in Andalusia (Spain). Data were gathered over 6 months on gender, age, care load, professional/family variables, and family-work balance, using a self-administered questionnaire. OSC and SCS scales were examined by using exploratory factorial analysis and Cronbach's α, and scores were compared by gender-stratified bivariate and multiple regression analyses. Intraclass correlation coefficients were calculated using a multilevel analysis. The response rate was 73.6%. We identified three OCS factors and two SCS factors. Lower scores were obtained by female versus male FPs in the OCS dimensions, but there were no gender differences in either SCS dimension.

Was access to health care easy for immigrants in Spain? The perspectives of health personnel in Catalonia and Andalusia.

Until April 2012, all Spanish citizens were entitled to health care and policies had been developed at national and regional level to remove potential barriers of access, however, evidence suggested problems of access for immigrants. In order to identify factors affecting immigrants' access to health care, we conducted a qualitative study based on individual interviews with healthcare managers (n=27) and professionals (n=65) in Catalonia and Andalusia, before the policy change that restricted access for some groups. A thematic analysis was carried out. Health professionals considered access to health care "easy" for immigrants and similar to access for autochthons in both regions. Clear barriers were identified to enter the health system (in obtaining the health card) and in using services, indicating a mismatch between the characteristics of services and those of immigrants. Results did not differ among regions, except for in Catalonia, where access to care was considered harder for users without a health card, due to the fees charged, and in general, because of the distance to primary health care in rural areas. In conclusion, despite the universal coverage granted by the Spanish healthcare system and developed health policies, a number of barriers in access emerged that would require implementing the existing policies. However, the measures taken in the context of the economic crisis are pointing in the opposite direction, towards maintaining or increasing barriers.

Ammonia Affects Astroglial Proliferation in Culture.

Primary cultures of rat astroglial cells were exposed to 1, 3 and 5 mM NH4Cl for up to 10 days. Dose- and time-dependent reductions in cell numbers were seen, plus an increase in the proportion of cells in the S phase. The DNA content was reduced in the treated cells, and BrdU incorporation diminished. However, neither ammonia nor ammonia plus glutamine had any effect on DNA polymerase activity. iTRAQ analysis showed that exposure to ammonia induced a significant reduction in histone and heterochromatin protein 1 expression. A reduction in cell viability was also noted. The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

[Familial characteristics and self-perceived health among female and male primary care physicians in Andalusia (Spain)]. / Las características familiares y la salud percibida en médicas y médicos de familia de Andalucía.

OBJECTIVE: To determine the relationships between a group of professional and family characteristics and the components of physical and mental health in female and male primary care physicians working in health centers in Andalusia (Spain). METHODS: A descriptive, cross-sectional, multicenter study was performed. The population consisted of urban health centers in Andalusia and their physicians. The sample comprised 88 health centers and 500 physicians. Measurements consisted of sex, age, professional characteristics (postgraduate training in family medicine, position of health center manager, accreditation as a residents' tutor, and workload based on patient quota and the mean number of patients/day); family responsibilities, defined by two dimensions of the family-work relationship (support overload-family support deficit and family-work conflict); and perceived physical and mental health. The data source was a self-administered questionnaire sent by surface mail. Multiple regression analyses were performed for physical and mental health for the whole sample and by gender. RESULTS: Responses were obtained from 368 physicians (73.6%). Mental health was worse in female physicians than in male physicians; no differences were found between genders in physical health. The family-work conflict was associated with physical and mental health in physicians of both genders. Physical health deteriorated with increasing age in both genders, improved in the female tutors of residents, and decreased with increasing family-work conflict in male physicians. Mental health decreased with increasing housework on the weekends and with family-work conflict in both genders. In male physicians, mental health deteriorated with postgraduate training in family medicine and improved if they were health center managers. CONCLUSIONS: Workload and professional characteristics have little relationship with the health of primary care physicians. Family characteristics play a greater role.

[Gender differences in the perception of professional achievement in family medicine, Spain]. / Diferencias de género en la percepción del logro profesional en especialistas de medicina familiar y comunitaria.

BACKGROUND: The concept of achievement is important to study the professional development. In medicine there are gender inequalities in career. The purpose was to know and compare the professional achievement's perceptions and attributions of female and male primary care physicians in Andalusia. METHOD: Qualitative study with 12 focus groups (October 2009 to November 2010). POPULATION: primary care physicians. SAMPLE: intentionally segmented by age, sex and health care management. Were conducted by sex: two groups with young physicians, two groups with middle aged and two with health care management. TOTAL: 32 female physician and 33 male physicians. Qualitative content analysis with Nuddist Vivo. RESULTS: Female and male physicians agree to perceive internal achievements and to consider aspects inherent to the profession as external achievements. The most important difference is that female physician related professional achievement with affective bond and male physician with institutional merit. Internal attributions are more important for female physician who also highlight the importance of family, the organization of working time and work-family balance. Patients, continuing education, institutional resources and computer system are the most important attributions for male physician. CONCLUSIONS: There are similarities and differences between female and male physicians both in the understanding and the attributions of achievement. The differences are explained by the gender system. The perception of achievement of the female physicians questions the dominant professional culture and incorporates new values in defining achievement. The attributions reflect the unequal impact of family and organizational variables and suggest that the female physicians would be changing gender socialization.

[Health impact assessment of the San Fernando street renewal project in Alcalá de Guadaíra (Seville, Spain)]. / Evaluación del impacto en la salud del proyecto de reurbanización de la calle San Fernando en Alcalá de Guadaíra (Sevilla).

OBJECTIVES: This study describes the design and implementation of a health impact assessment (HIA) conducted in 2010 of the regeneration project of San Fernando Street, the main avenue crossing the San Miguel-El Castillo neighborhood in Alcala de Guadaíra (Seville, Spain). This project is part of the wider URBAN Plan aimed at the social, urban and economic regeneration of the city's historic center. METHODS: This experience followed the standard HIA stages and procedures. The review of published evidence was complemented with new qualitative information gathered by means of a participative workshop with the local population and interviews with social and health workers involved in the neighborhood. RESULTS: During the building stage of the project, the adverse impacts were related to a worsening of the air quality, increased noise pollution, mobility restrictions and a higher risk of accidents, particularly among older or disabled people. Once the building stage was finished, the health benefits were associated with significant improvements in physical accessibility and the population's access to health services and other goods and services. Other positive effects were the enhanced safety and attractiveness of the neighborhood and the new opportunities for socializing, social cohesion and increasing the community's self-esteem. CONCLUSIONS: This is the first HIA experience in Andalusia whose results have been integrated into a formal cycle of decision making in the local community. This experience has provided new evidence of the potential of HIA and its applicability and acceptance at the municipal level and has has also facilitated a learning process and the piloting of new methods and tools associated with the HIA process.