Severe perioperative anaphylaxis: Incidence in a tertiary hospital in Spain over a 20-year period. A historical cohort study.

OBJECTIVE: To assess the incidence of severe perioperative anaphylaxis, the mechanisms involved, the value of laboratory/skin tests, and the most effective treatments. METHODS: A historical cohort study conducted in a tertiary public hospital in Spain. Patients that had undergone anaesthesia during the 20-year period were included. In these patients, 66 cases of severe anaphylaxis were found. In patients with suspicion of severe anaphylaxis, levels of blood histamine at less than 15min and serum tryptase at 2, 6, and 24h following the reaction were determined. Skin and specific IgE tests were performed between 4 and 8 weeks later. RESULTS: Over the 20-year period, 288 594 anaesthetic procedures were performed. We observed cases of 66 severe anaphylaxis reaction (59% men; age, 60.8±17.3 years. Symptoms observed were cardiovascular (86%), respiratory (73%), and mucocutaneous (56%). Elevated serum tryptase levels were associated with degree of severity at 2 (P<.0001) and 6h (P=.026) and were highest in IgE-mediated reactions (P=.020). All patients required treatment, and 3 events were fatal. In 84.8% of patients, skin and/or specific IgE tests were positive for antibiotics (35.8%), non-steroidal anti-inflammatory drugs (23.1%), neuromuscular blocking agents (15.4%) and latex (15.4%). CONCLUSIONS: The incidence of severe anaphylaxis in our hospital was 1 in 4.373 anaesthetic procedures, with a death rate of 4.5%. All cases required treatment. Serum tryptase was a good predictor of reaction severity. The most frequent causative agents were antibiotics, non-steroidal anti-inflammatory drugs, neuromuscular blocking agents and latex.

Anafilaxia perioperatoria severa: Incidencia en un hospital terciario en España durante 20 años. Estudio de cohorte histórico / Severe perioperative anaphylaxis: Incidence in a tertiary hospital in Spain over a 20-year period. A historical cohort study

Objetivo: Determinar la incidencia de reacciones perioperatorias graves, los mecanismos implicados, los tratamientos realizados y la utilidad del protocolo diagnóstico propuesto. Métodos: Estudio de cohorte histórico en un hospital público terciario en España. Se incluyeron pacientes que recibieron anestesia durante 20 años. En pacientes con sospecha de anafilaxia grave, se determinaron los niveles de histamina en sangre en menos de 15 min y triptasa sérica a las 2, 6 y 24 h después de la reacción. Se realizaron pruebas cutáneas e IgE específica a las 4-8 semanas. Resultados: Durante el período de 20 años, se realizaron 288.594 procedimientos anestésicos. Observamos 66 reacciones de anafilaxia grave (59% hombres; edad: 60,8 ± 17,3 años). Los síntomas fueron cardiovasculares (86%), respiratorios (73%) y mucocutáneos (56%). Los niveles elevados de triptasa sérica se asociaron con un mayor nivel de gravedad a las 2 (p < 0,0001) y 6 h (p = 0,026) y fueron más elevados en las reacciones IgE mediadas (p = 0,020). Todos los pacientes requirieron tratamiento y la muerte ocurrió en tres casos. En el 84,8% de los pacientes las pruebas cutáneas y/o IgE específicas fueron positivas a antibióticos (35,8%), antiinflamatorios no esteroideos (23,1%), bloqueantes neuromusculares (15,4%) y látex (15,4%). Conclusiones: La incidencia de anafilaxia grave en nuestro hospital fue 1:4.373 procedimientos anestésicos, con una mortalidad del 4,5%. Todos los casos requirieron tratamiento. La triptasa sérica fue un buen predictor de la gravedad de la reacción. Los agentes etiológicos más frecuentes fueron antibióticos, antiinflamatorios no esteroideos, bloqueantes neuromusculares y látex.(AU)

Objective: To assess the incidence of severe perioperative anaphylaxis, the mechanisms involved, the value of laboratory/skin tests, and the most effective treatments. Methods: A historical cohort study conducted in a tertiary public hospital in Spain. Patients that had undergone anaesthesia during the 20-year period were included. In these patients, 66 cases of severe anaphylaxis were found. In patients with suspicion of severe anaphylaxis, levels of blood histamine at less than 15 minutes and serum tryptase at 2, 6, and 24 hours following the reaction were determined. Skin and specific IgE tests were performed between 4 and 8 weeks later. Results: Over the 20-year period, 288 594 anaesthetic procedures were performed. We observed cases of 66 severe anaphylaxis reaction (59% men; age, 60.8 ± 17.3 years. Symptoms observed were cardiovascular (86%), respiratory (73%), and mucocutaneous (56%). Elevated serum tryptase levels were associated with degree of severity at 2 (p < 0.0001) and 6 hours (p = 0.026) and were highest in IgE-mediated reactions (p = 0.020). All patients required treatment, and 3 events were fatal. In 84.8% of patients, skin and/or specific IgE tests were positive for antibiotics (35.8%), non-steroidal anti-inflammatory drugs (23.1%), neuromuscular blocking agents (15.4%) and latex (15.4%). Conclusions: The incidence of severe anaphylaxis in our hospital was 1 in 4 373 anaesthetic procedures, with a death rate of 4.5%. All cases required treatment. Serum tryptase was a good predictor of reaction severity. The most frequent causative agents were antibiotics, non-steroidal anti-inflammatory drugs, neuromuscular blocking agents and latex.(AU)

Confident Perception of Primary Care Physicians Correlates to the Attitude Toward Donation and Organ Transplantation: A Multicenter Study of Medical And Nursing Spanish Students.

A primary care physician (PCP) not only accompanies the patient in the process of an illness, but throughout his or her life. The confidence we have in these health professionals is fundamental, and their favorable attitude toward organ donation and transplantation (ODT) has a significant influence on the population. OBJECTIVE: To analyze trust in PCPs among Spanish medical and nursing students, the relationship with their attitude toward ODT, and the factors that condition it. METHODS AND DESIGN: A sociologic, multicenter, and observational study. POPULATION: medical and nursing students in Spanish universities. DATABASE: Collaborative International Donor Project, stratified by geographic area and academic course. A validated questionnaire (PCID-DTO-RIOS) was self-administered and completed anonymously. A sample of 9598 medical and 10,566 nursing students (99% confidence and precision of ±1%), stratified by geographic area and year of study. RESULTS: Completion rate: 90%. With respect to students' trust in their physician, 18% (n = 3267) of them totally trust (completely), 45% (n = 8101) trust enough, 30% (n = 5478) of them have not enough trust, and 7% not at all. Comparing groups, medical students totally trust more in PCPs than nursing students (55% vs 45%; P < .000), however, nursing students have less than enough trust in their PCP than medical students (53% vs 47%; P < .000). Students that totally trust in their PCP were more in favor toward ODT than students with not enough trust (83% vs 77%; P < .000). CONCLUSION: Only 18% of Spanish medical and nursing students totally trust in their PCP. Attitude toward ODT is related to a higher level of trust in PCPs among these students.

Information About Donation and Organ Transplantation Among Spanish Medical Students.

INTRODUCTION: Health professionals are asked to promote health, especially organ transplantation; however, they do not always have specific training. OBJECTIVE: To analyze information about donation and organ transplantation among Spanish medical students. METHOD: The population under study is medical students in Spanish universities using the database of the International Donor Collaborative Project, stratified by geographic area and academic year (n = 9275). The instrument used is the attitude questionnaire for organ donation for "PCID-DTO-Ríos" transplantation, validated with an explained variance of 63.203% and α = 0.834. The Student t test was applied together with the χ2 test, complemented by an analysis of the remainders, and Fisher's exact test was applied. RESULTS: Of the students, 74% indicate that they have received information from university professors about organ transplant. Concerning specific issues with the donation, it is notable that only 66.7% (n = 6190) know and accept the concept of brain death as the death of a person. However, only 22% consider themselves as having good information, and 35.3% indicate that their information is scarce or void. Students indicate having received information about transplant from other extra-university sources, such as television and Internet (80.9%), books and magazines (73.2%), and the press (66.9%). From the information obtained in the sociofamilial field, 60.7% have obtained information from the family and 58.1% from friends. Of this information, 9% has been negative from friends, 7.5% from family, 6% from the Internet and television, and 4% from university professors. CONCLUSION: Spanish medical students believe they have little information about organ transplantation and have received negative information.

Are Medical Students of Non-Spanish Nationality Studying in Spain as Sensitized to Transplantation as Those of Spanish Nationality?

INTRODUCTION: Awareness of organ donation among Spanish doctors and medical students is very positive. However, the emerging group of professionals of non-Spanish nationality studying in Spain has not been analyzed. OBJECTIVE: To analyze the differences in the attitudes toward the different types of donation among medical students, according to their nationality. METHODS: The population under study is medical students in Spanish universities using the database of the International Collaborative Donor Project, stratified by geographic area and academic year. Groups under study include students of non-Spanish nationality as group 1 (n = 1570) and students of Spanish nationality as Group 2 (n = 7705). Instruments are validated questionnaires of attitude toward donation "PCID-DTO-Ríos," "PCID-DVR-Ríos," "PCID-DVH-Ríos," and "PCID-XenoTx-Ríos." RESULTS: The attitude toward the donation of own organs after death is similar in both groups (P = .703). Non-Spaniards are 79.2% in favor compared to 79.6% of Spaniards. Living kidney donation, both unrelated (33.3% vs 29.3% in favor; P = .001) and related (91.2% vs 89, 6% in favor; P = .047), is more favorable among non-Spanish students. There are no differences regarding non-related living liver donation (29.7% vs 29.3% in favor; P = .063), but there are differences in the results for related living liver donation (94.1% vs 88%; P < .001). The attitude toward xenotransplantation of organs is similar (80.8% vs 80.8%; P = .999). CONCLUSIONS: Awareness of the donation of organs among Spanish medical students is similar to non-Spanish students studying in Spain, except the attitude toward living donation.

What Reasons Lead Spanish Medical Students Not to Be in Favor of Organ Donation?

INTRODUCTION: Despite sensitization of medical students toward the donation of organs, a non-negligible percentage of students are not in favor of donation. OBJECTIVE: To analyze the reasons of Spanish medical students who do not have a favorable attitude toward the donation of their own organs after death. METHOD: The population under study is medical students in Spanish universities, using the database of the International Donor Collaborative Project, stratified by geographic area and academic year. The questionnaire completion was anonymous and self-administered. The study group is medical students with an unfavorable attitude toward organ donation. The assessment instrument used is a validated questionnaire of attitude toward the donation of organs for transplant, "PCID-DTO-Ríos." The reasons against the donation are valued in the questionnaire through a question. RESULTS: Of the participants included in the PCID, students who are not in favor of organ donation were selected (n = 1899). Of them, 8.1% (n = 154) are against and 91.1% (n = 1745) are doubtful. The main reasons indicated are the fear of apparent death in 11.4% of respondents, fear of possible mutilation after donation in 11.1%, and religious reasons in 2.6%. Of those, 6.9% indicate other reasons but do not clearly specify the reasons, using words such as "fear" (2.5%) or "doubts about the process" (4.1%); 66.2% (n = 1257) indicated an "assertive refusal" ("I don't want to express my reasons"). CONCLUSION: Twenty percent of Spanish medical students are not in favor of donating their organs and are not in favor of showing their reasons.

LOXL2-mediated H3K4 oxidation reduces chromatin accessibility in triple-negative breast cancer cells.

Oxidation of H3 at lysine 4 (H3K4ox) by lysyl oxidase-like 2 (LOXL2) generates an H3 modification with an unknown physiological function. We find that LOXL2 and H3K4ox are higher in triple-negative breast cancer (TNBC) cell lines and patient-derived xenografts (PDXs) than those from other breast cancer subtypes. ChIP-seq revealed that H3K4ox is located primarily in heterochromatin, where it is involved in chromatin compaction. Knocking down LOXL2 reduces H3K4ox levels and causes chromatin decompaction, resulting in a sustained activation of the DNA damage response (DDR) and increased susceptibility to anticancer agents. This critical role that LOXL2 and oxidized H3 play in chromatin compaction and DDR suggests that functionally targeting LOXL2 could be a way to sensitize TNBC cells to conventional therapy.

Do Religious Factors Influence the Attitude Toward Organ Donation Among Medical Students? A Spanish Multicenter Study.

INTRODUCTION: Religious factors have conditioned the attitude toward organ donation and transplantation (ODT) since the beginning of transplantation, despite the fact that most religions are in favor of transplantation. OBJECTIVE: To assess the impact of religious beliefs of medical students on their attitude toward ODT. METHOD: Population under study: Medical students in Spanish universities. STUDY SAMPLE: Stratified by geographical area and academic course. Assessment instrument: Attitude ODT questionnaire PCID-DTO-Ríos, anonymous and self-administered. RESULTS: Of all students, 42% (n = 3907) declare themselves atheists or agnostics. The remaining 58% (n = 5368) declare themselves to be religious, the majority being Catholic (55%, n = 5102). Of the rest, 0.2% are Muslims (n = 8), 0.1% Protestants (n = 1), and the remaining 2.7% (n = 257) indicate other religious doctrines but do not want to specify it. Regarding their attitude toward ODT, those who consider themselves atheists or agnostics have a more favorable attitude than those who consider themselves religious (84% versus 76%; P < .001). Among those who follow some kind of religion, Catholics are more in favor of ODT than non-Catholics (77% vs 64%, P < .001). Note that among the religious, only 57% (n = 3050) know which religion is in favor of transplantation, while 22% (n = 1,152) consider that it has not been pronounced on the matter, 13% (n = 723) think the religion is against donation, and the remaining 8% (n = 443) do not know. CONCLUSION: The religion professed by medical students conditions their attitude toward donation, with the atheists and agnostics being more in favor of donation.

Are High Rates of Deceased Donation Conditioning the Attitude Toward Liver Donation Among Medical Students?

The involvement of health professionals from their training period is important for the promotion of living liver donation. There are data that indicate that the awareness of living donation is lower in areas with high rates of deceased donation. OBJECTIVE: To analyze the attitude toward living liver donation among Spanish medical students, according to donation rates of their regions. METHOD: Population under study: Medical students in Spanish universities. Database of the Collaborative International Donor Project, stratified by geographic area and academic course. The completion was anonymous and self-administered. Groups under study: group 1 (n = 1136): students in universities of regions with >50 donors per million population (pmp); group 2 (n = 2018): students in region universities with <40 donors pmp. Assessment instrument: the attitude questionnaire for living liver donation Proyecto Colaborativo Internacional Donante sobre Donación de Vivo Hepático-Ríos (PCID-DVH Ríos). RESULTS: The attitude toward related liver donation is more favorable among the students of regions with <40 donors pmp than among those of >50 donors pmp. Thus, in group 1, a total of 88% (n = 1002) of students are in favor compared with 91% (n = 1831) of group 2 (P=.02). The psychosocial profile of each study group about their attitude toward living related liver donation is analyzed. There is a similar profile between the 2 groups, although there are differences in some variables such as age, a belief that one might need a transplant, family discussion about donation and transplantation, discussion with friends about donation and transplantation, and knowing about a donor. CONCLUSIONS: The awareness of living related donation among Spanish medical students is greater among the regions with lower organ donation rates.

Sensitization Toward Organ Donation Among Medical Students in Spanish Regions With More Than 50 Donors Per Million Population.

The awareness of organ donation among health professionals is important at the time of transplant promotion. In this sense, the training and awareness of the professionals in training is fundamental. OBJECTIVE: To analyze the differences in the attitude toward organ donation and the factors that condition it among medical students of regions with donation rates >50 donors per million population (pmp) with respect to those with rates <40 donor pmp. METHOD: Population under study: medical students in Spanish universities. Database of the Collaborative International Donor Project, stratified by geographic area and academic course. The completion was anonymous and self-administered. Groups under study: Group 1 (n = 1136): students in universities of regions with >50 donors pmp. Group 2 (n = 2018): university students in regions with <40 donors pmp. Assessment instrument: attitude questionnaire for organ donation for transplant PCID-DTO-Ríos. RESULTS: The attitude toward organ donation for transplantation is similar among students from the autonomous communities with >50 donors pmp and with <40 donors pmp. In group 1, 79% (n = 897) of students are in favor compared with 81% (n = 1625) of group 2 (P=.29). The psychosocial profile toward donation is similar in both groups relating to the following variables (P < .05): sex, having discussed transplantation with family and as a couple, considering the possibility of needing a transplant, involvement in prosocial activities, attitude toward the manipulation of corpses, knowledge of the brain death concept, and religion. CONCLUSIONS: The awareness of organ donation in Spanish medical students is quite homogeneous and is not related to the local donation rates of each region.

A Multicentre and stratified study of the attitude of medical students towards organ donation in Spain.

INTRODUCTION: Medical students represent a new generation of medical thought, and if they have a favourable attitude towards organ donation this will greatly encourage its promotion. OBJECTIVE: To analyse the attitude of medical students in Spanish universities towards the donation of their own organs and to determine the factors affecting this attitude. MATERIAL AND METHODS: Type of study: A sociological, interdisciplinary, multicentre, and observational study in Spain. STUDY POPULATION: Students studying a degree in medicine enrolled in Spain (n = 34,000). SAMPLE SIZE: A sample of 9598 students (confidence of 99% and precision of ±1%), stratified by geographical area and academic year. Instrument of measurement: A validated questionnaire of attitude towards organ donation and transplantation (PCID-DTO RIOS) was self-administered and completed anonymously. RESULTS: The questionnaire completion rate was 95.7% (n = 9.275). 80% were in favour of donation, 2% against and 18% were undecided. The following main variables were related to a favourable attitude: being of the female sex (Odds Ratio = 1.739); being in the sixth year of the degree (OR = 2.506); knowing a donor (OR = 1.346); having spoken about the subject with one's family (OR = 2.132) and friends (OR = 1.333); having a family circle that is in favour, more specifically, having a father (OR = 1.841), mother (OR = 2.538) or partner in favour (OR = 2.192); being a blood donor (OR = 2.824); acceptance of the mutilation of the body if it were necessary (OR = 2.958); and being an atheist or an agnostic (OR = 1.766). CONCLUSIONS: Spanish medical students generally have a favourable attitude towards organ donation, although 20% are not in favour.

Do Spanish Medical Students Understand the Concept of Brain Death?

OBJECTIVE: To analyze the level of understanding of the brain death concept among medical students in universities in Spain. METHODS: This cross-sectional sociological, interdisciplinary, and multicenter study was performed on 9598 medical students in Spain. The sample was stratified by geographical area and academic year. A previously validated self-reported measure of brain death knowledge (questionnaire Proyecto Colaborativo Internacional Donante sobre la Donación y Transplante de Organos) was completed anonymously by students. RESULTS: Respondents completed 9275 surveys for a completion rate of 95.7%. Of those, 67% (n = 6190) of the respondents understood the brain death concept. Of the rest, 28% (n = 2652) did not know what it meant, and the remaining 5% (n = 433) believed that it did not mean that the patient was dead. The variables related to a correct understanding of the concept were: (1) being older ( P < .001), (2) studying at a public university ( P < .001), (3) year of medical school ( P < .001), (4) studying at one of the universities in the south of Spain ( P = .003), (5) having discussed donation and transplantation with the family ( P < .001), (6) having spoken to friends about the matter ( P < .001), (7) a partner's favorable attitude toward donation and transplantation ( P < .001), and (8) religious beliefs ( P < .001). CONCLUSIONS: Sixty-seven percent of medical students know the concept of brain death, and knowledge improved as they advanced in their degree.

Transcriptome analysis of severely active chronic spontaneous urticaria shows an overall immunological skin involvement.

BACKGROUND: The knowledge about chronic spontaneous urticaria (CSU) phenotypes is based on its clinical characteristics, associated comorbidities, course of the disease, and its response to the available effective drugs. Genotype expression and its further correlation with CSU phenotypes are still unknown. We describe the cutaneous transcriptome of patients suffering a severely active CSU refractory to antihistamine treatment. METHODS: Through the bioinformatic analysis of the whole Human Genome with Oligo Microarrays and quantitative real-time polymerase chain reaction (qPCR), relevant genes expressed in nonlesional (NLS-CSU) and lesional skin (LS-CSU) and peripheral blood were identified in 20 patients suffering from severely active CSU and 10 healthy controls (HCs). RESULTS: From 39 genes differentially expressed in NLS-CSU when compared with HCs, 31 (79.48%) were confirmed by qPCR corresponding to genes involved in epidermal homeostasis and dermal repair. From the analysis comparing LS-CSU with NLS-CSU, a selection of 142 genes was studied with qPCR, and 103 (72.53%) were confirmed. Differentially expressed genes in the phenomenon of wheal development are involved in a variety of biological functions as, epidermal differentiation, intracellular signal function, transcriptional factors cell cycle differentiation, inflammation, or coagulation. Differentially expressed genes that uniformly increase or decrease along the skin worsening until the wheal appearance is shown. CONCLUSION: The skin of CSU patients with a severely active disease shows an overall immunological skin involvement showing a peculiar gene profile.

Medical students faced with related and unrelated living kidney donation: a stratified and multicentre study in Spain.

INTRODUCTION: The attitude of medical students towards living kidney donation (LKD) is of great interest given that they will become promoters of this technique in the near future. OBJECTIVE: To analyse the attitude of Spanish medical students towards related and unrelated LKD and to determine the factors affecting this attitude. MATERIALS AND METHODS: Type of study: A sociological, interdisciplinary, multicentre, and observational study. STUDY POPULATION: Medical students enrolled in Spain (n = 34.000). SAMPLE SIZE: A sample of 9598 students (99 % confidence and precision of ±1 %), stratified by geographical area and academic year. Measurement instrument: A validated questionnaire (PCID-DVR RIOS) was administered and completed anonymously. RESULTS: There was a completion rate of 95.7 % (n = 9275); 93 % (n = 8630) were in favour of related LKD, and 30 % (n = 2784) were in favour of unrelated LKD. The following factors were associated with this attitude: (1) age (p = 0.008); (2) sex (p < 0.001); (3) year of university degree (p < 0.001); (4) a belief that a transplant might be necessary in the future (p < 0.001); (5) attitude towards deceased organ donation (p < 0.001); (6) a willingness to accept a kidney from a living donor (p < 0.001); (7) attitude towards living liver donation (p < 0.001); (8) a partner's attitude towards donation (p < 0.001); (9) having spoken about the subject with one's family (p < 0.001), or friends (p < 0.001); (10) pro-social behaviour (p < 0.001); (11) the respondent's religious attitude (p < 0.001); and (12) fear of possible mutilation of the body after donation (p < 0.001). CONCLUSIONS: The attitude of medical students towards LKD is very favourable when it is the related kind of donation, and it is associated with factors of general knowledge about organ donation and transplantation and social interaction and religion.

Circulating NK-cell subsets in renal allograft recipients with anti-HLA donor-specific antibodies.

Detection of posttransplant donor-specific anti-HLA antibodies (DSA) constitutes a risk factor for kidney allograft loss. Together with complement activation, NK-cell antibody-dependent cell mediated cytotoxicity (ADCC) has been proposed to contribute to the microvascular damage associated to humoral rejection. In the present observational exploratory study, we have tried to find a relationship of circulating donor-specific and non donor-specific anti-HLA antibodies (DSA and HLA non-DSA) with peripheral blood NK-cell subsets and clinical features in 393 renal allograft recipients. Multivariate analysis indicated that retransplantation and pretransplant sensitization were associated with detection of posttransplant DSA. Recipient female gender, DR mismatch and acute rejection were significantly associated with posttransplant DSA compared to HLA non-DSA. In contrast with patients without detectable anti-HLA antibodies, DSA and HLA non-DSA patients displayed lower proportions of NK-cells, associated with increased CD56(bright) and NKG2A(+) subsets, the latter being more marked in DSA cases. These differences appeared unrelated to retransplantation, previous acute rejection or immunosuppressive therapy. Although preliminary and observational in nature, our results suggest that the assessment of the NK-cell immunophenotype may contribute to define signatures of alloreactive humoral responses in renal allograft recipients.

PARP-2 sustains erythropoiesis in mice by limiting replicative stress in erythroid progenitors.

Erythropoiesis is a tightly regulated process in which multipotential hematopoietic stem cells produce mature red blood cells. Here we show that deletion of poly(ADP-ribose) polymerase-2 (PARP-2) in mice leads to chronic anemia at steady state, despite increased erythropoietin plasma levels, a phenomenon not observed in mice lacking PARP-1. Loss of PARP-2 causes shortened lifespan of erythrocytes and impaired differentiation of erythroid progenitors. In erythroblasts, PARP-2 deficiency triggers replicative stress, as indicated by the presence of micronuclei, the accumulation of γ-H2AX (phospho-histone H2AX) in S-phase cells and constitutive CHK1 and replication protein A phosphorylation. Transcriptome analyses revealed the activation of the p53-dependent DNA-damage response pathways in PARP-2-deficient cells, culminating in the upregulation of cell-cycle and cell death regulators, concomitant with G2/M arrest and apoptosis. Strikingly, while loss of the proapoptotic p53 target gene Puma restored hematocrit levels in the PARP-2-deficient mice, loss of the cell-cycle regulator and CDK inhibitor p21 leads to perinatal death by exacerbating impaired fetal liver erythropoiesis in PARP-2-deficient embryos. Although the anemia displayed by PARP-2-deficient mice is compatible with life, mice die rapidly when exposed to stress-induced enhanced hemolysis. Our results pinpoint an essential role for PARP-2 in erythropoiesis by limiting replicative stress that becomes essential in the absence of p21 and in the context of enhanced hemolysis, highlighting the potential effect that might arise from the design and use of PARP inhibitors that specifically inactivate PARP proteins.

Loss of PIDD limits NF-κB activation and cytokine production but not cell survival or transformation after DNA damage.

Activation of NF-κB (nuclear factor of kappa light chain gene enhancer in B cells) in response to DNA damage is considered to contribute to repair of genetic lesions, increased cell survival and cytokine release. The molecular mechanisms orchestrating this cytoplasmic event involve core components of the nuclear DNA damage response machinery, including ATM-kinase (ataxia telangiectasia mutated kinase) and PARP-1 (poly (ADP-ribose) polymerase 1). The physiological consequences of defective NF-κB activation in this context, however, remain poorly investigated. Here we report on the role of the 'p53-induced protein with a death domain', PIDD, which appears rate limiting in this process, as is PARP-1. Despite impaired NF-κB activation, DNA damage did not increase cell death or reduce clonal survival of various cell types lacking PIDD, such as mouse embryonic fibroblasts or stem and progenitor cells of the hematopoietic system. Furthermore, lymphomagenesis induced by γ-irradiation (IR) was unaffected by deficiency for PIDD or PARP-1, indicating that loss of DNA damage-triggered NF-κB signalling does not affect IR-driven tumorigenesis. However, loss of either gene compromised cytokine release after acute IR injury. Hence, we propose that NF-κB's most notable function after DNA damage in primary cells is related to the release of cytokines, thereby contributing to sterile inflammation.

Nuclear PARP-1 protein overexpression is associated with poor overall survival in early breast cancer.

BACKGROUND: Poly(ADP-ribose)polymerase-1 (PARP-1) is a highly promising novel target in breast cancer. However, the expression of PARP-1 protein in breast cancer and its associations with outcome are yet poorly characterized. PATIENTS AND METHODS: Quantitative expression of PARP-1 protein was assayed by a specific immunohistochemical signal intensity scanning assay in a range of normal to malignant breast lesions, including a series of patients (N = 330) with operable breast cancer to correlate with clinicopathological factors and long-term outcome. RESULTS: PARP-1 was overexpressed in about a third of ductal carcinoma in situ and infiltrating breast carcinomas. PARP-1 protein overexpression was associated to higher tumor grade (P = 0.01), estrogen-negative tumors (P < 0.001) and triple-negative phenotype (P < 0.001). The hazard ratio (HR) for death in patients with PARP-1 overexpressing tumors was 7.24 (95% CI; 3.56-14.75). In a multivariate analysis, PARP-1 overexpression was an independent prognostic factor for both disease-free (HR 10.05; 95% CI 5.42-10.66) and overall survival (HR 1.82; 95% CI 1.32-2.52). CONCLUSIONS: Nuclear PARP-1 is overexpressed during the malignant transformation of the breast, particularly in triple-negative tumors, and independently predicts poor prognosis in operable invasive breast cancer.

Local identification of porcine haptoglobin in salivary gland and diaphragmatic muscle tissues.

In order to clarify the origin of the haptoglobin (Hp) quantified in saliva and meat juice samples, the extrahepatic localization of Hp in salivary gland and in diaphragmatic muscle, as part of the systemic acute phase response in pigs, was studied by immunohistochemistry. For this purpose a specific monoclonal antibody (mAb) produced by immunising mice with purified porcine Hp was used. Reactivity of the mAb was assessed by direct ELISA and by western blot, which showed the ability and specificity of the mAb to identify porcine haptoglobin as a purified antigen or in porcine serum in a native or denatured but non-reduced state. Five healthy and five diseased pigs were sampled at slaughter for serum and tissue procurement. Hepatic immunohistochemical analysis was used as control of the acute phase reaction status. In the liver, cell immunostaining revealed a perinuclear, cytoplasmic localization of Hp within hepatocytes, following mainly a periacinar pattern. Extrahepatic immunohistochemical analysis revealed positive cells in the glandular acini and duct epithelial cells of the salivary gland and intrasarcoplasmic immunolabelling of random diaphragmatic myofibers. A possible role of both salivary gland and diaphragmatic muscle on local Hp production could be postulated based on the present immunohistochemical study, which supports the concept that other cells besides hepatocytes may have the potential to produce Hp in the pig.