A mutation in the Z-line Cypher/ZASP protein is associated with arrhythmogenic right ventricular cardiomyopathy.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an important cause of malignant arrhythmia and sudden death particularly in young people. Although it is considered a desmosomal disease, mutations in non-desmosomal genes have also been identified. We report on a family where a mutation in LDB3 is associated with this condition. The index case and first and second degree relatives underwent a complete clinical evaluation: physical examination, electrocardiography (ECG), signal-averaged ECG, 2D echocardiogram, cardiac magnetic resonance and 24-h monitoring. After ruling out mutations in the five desmosomal genes, genetic testing by means of Next Generation Sequencing was carried out on the proband. A heterozygous missense mutation in LDB3 c.1051A>G was identified. This result was confirmed by subsequent Sanger DNA sequencing. Another six carriers were identified amongst her relatives. Three subjects fulfilled the criteria for a definitive diagnosis of ARVC and one reached a borderline diagnosis. In conclusion, this is the first family with ARVC where a mutation in LDB3 is associated with ARVC. Next generation sequencing arises as a particular useful tool to point to new causative genes in ARVC.

Disability, quality of life, personality, cognitive and psychological variables associated with fatigue in patients with multiple sclerosis.

OBJECTIVE: To examine the associations between function, quality of life, personality, cognitive and psychological outcomes with fatigue in patients with MS. METHODS: One hundred and eight patients (54% women) with definite MS participated. MS-related fatigue was assessed with the Fatigue Impact Scale (FIS). Demographic and clinical data (weight, height, medication and history of pain), specific disease outcomes (Functional Assessment of Multiple Sclerosis/FAMS), general disease outcomes (Beck Depression Inventory/BDI-II, and Short-Form Health Survey 36/SF-36) and personality (NEO Five-Factor Inventory/NEOFFI) were assessed. Correlation and regression analyses were performed to determine associations between variables. RESULTS: A significant positive correlation existed between the FIS and EDSS (r=0.190; P<0.05). Significant negative correlations between the FIS with specific disease (FAMS mobility: r=-0.333, P<0.01; FAMS symptoms: r=-0.443, P<0.01; FAMS emotional well-being: r=-0.412, P<0.01; FAMS general contentment: r=-0.325, P<. 01; and, FAMS thinking/fatigue: r=-0.706, P<0.01); general disease (all domains SF36: -0.508

[Donation protocol following controlled cardiac death (Maastricht type III donation). First experience]. / Protocolo de donación tras la muerte cardiaca controlada (donante tipo iii de Maastricht). Experiencia inicial.

OBJECTIVE: To present our experience with the implementation of a donation protocol following controlled cardiac death (Maastricht type III donation). DESIGN: A retrospective descriptive and observational study was made. SETTING: Intensive Care Unit of a third-level university hospital. PATIENTS: Eight patients in an irreversible state, in which withdrawal of all life support had been agreed, were evaluated as potential donors. INTERVENTIONS: Application of the adopted protocol. VARIABLES OF INTEREST: Clinical data of donors, evaluation of a donation protocol following cardiac death, warm ischemia times, and short-term outcome of the recipients. RESULTS: Eight patients were evaluated. In one case donation was not possible because no cardiac arrest developed in the 120 minutes after extubation. The 7 remaining patients were effective kidney donors. Warm ischemia times were less than 23 minutes in all cases. Although 7 of the 14 recipients suffered delayed graft function, all of them achieved good renal function. CONCLUSION: Donation after cardiac death in patients in an overwhelming and irreversible state represents a potential source of donors not previously considered in this country. The prior development of a consensus-based protocol can help increase the number of organs in combination with those obtained after brain death. In our experience, the results of kidney transplants obtained from donors after cardiac death are good, and the success of these types of protocols could be extended to other organs such as the liver and lungs.

Fractional calculus as a generalized kinetic model for biochemical methane potential tests.

This study presents a fractional calculus model as a generalized kinetic model for estimating the maximum methane yield and degradation kinetics in biomethane potential (BMP) assays, a key analytical method in anaerobic digestion research and application. The fractional model outperformed common first-order kinetic models by yielding superior data fitting and properly managing substrate heterogeneity. The fractional model showed robust performance in mono-digestion, co-digestion and pre-treatment BMP assays with or without presence of large tailing or sigmoidal patterns in the BMP curve. The main advantage of the fractional model over other models is its ability to capture the complexities of the methane production process without losing model accuracy. Assessment of the mathematical model revealed that for fractional orders greater than 0.8 the Mittag-Leffler sequence could be transformed into a more computationally efficient exponential function.

Hematological and biochemical blood values in wild populations of black howler monkeys (Alouatta pigra) of Campeche, México.

BACKGROUND: The clinical analyses of free-ranging New World primates are extremely limited. The aim of this study was to obtain baseline data of blood chemistry and hematological values of free-ranging Alouatta pigra. METHOD: 17 males and 17 females were captured. We collected blood from the caudal tail vein; conventional techniques were used to analyze the samples RESULTS: Hemoglobin, hematocrit, and erythrocytes were significantly higher in males than in females. In relation to biochemical blood values, albumin/globulin ratio, cholesterol, triglycerides, calcium, phosphorus, magnesium, and iron were significantly higher in males than in females, while creatinine was significantly higher in females than in males. CONCLUSIONS: In general, measured parameters are similar to those of other primates and will be used as reference values for futures studies.

Role of normothermic perfusion with ECMO in donation after controlled cardiac death in Spain.

Spain has become one of the most active countries in donation after controlled cardiac death, using normothermic abdominal perfusion with ECMO in more than 50% of all donors - a situation contributed to by the creation of mobile teams to support hospitals lacking this technology. The donation process must be respectful of the wishes and values of the patients and their relatives, especially if there is pre mortem manipulation, and the absence of cerebral perfusion should be guaranteed. The liver is the most benefited organ by reducing biliary complications as well as the loss of grafts. In renal transplantation, the technique could contribute to reduce the incidence of delayed graft function. In addition, the procedure is compatible with surgical rapid recovery in hypothermia when there is also lung donation. The future lies in the consolidation of cardiac donation by extending normothermic perfusion to the thoracic cavity.

Enhancement of the intensity, persistence, and passive transfer of delayed-type hypersensitivity lesions by pertussigen in mice.

Pertussigen, a purified protein from Bordetella pertussis, was shown to increase delayed-type hypersensitivity (DTH) to protein antigens in mice. First, it caused an approximately twofold enhancement of the magnitude of 24-h DTH reactions. Second, the peak magnitude of DTH was delayed to 4-7 d after challenge, at which time it was five times more intense than in mice not receiving pertussigen. This reaction was antigen specific, and histologically was characterized by a dense mononuclear infiltrate. Third, pertussigen prolonged DTH so that it was still detectable 3-6 wk after challenge. The effect of pertussigen was seen only in antigen-driven reactions and was time and dose dependent, with 400 ng given 3 d after immunization resulting in the most prolonged reaction. The administration of pertussigen to the recipients of sensitized lymph node cells resulted in DTH that was more intense and prolonged than the reactions in control mice. Administration of pertussigen provides a model of prolonged and enhanced T cell-dependent inflammatory responses.

Role of normothermic perfusión with ECMO in donation after controlled cardiac death in Spain. / Papel de la perfusión normotérmica con oxigenación de membrana extracorporea en la donación en asistolia controlada en España.

Spain has become one of the most active countries in donation after controlled cardiac death, using normothermic abdominal perfusion with ECMO in more than 50% of all donors - a situation contributed to by the creation of mobile teams to support hospitals lacking this technology. The donation process must be respectful of the wishes and values of the patients and their relatives, especially if there is pre mortem manipulation, and the absence of cerebral perfusion should be guaranteed. The liver is the most benefited organ by reducing biliary complications as well as the loss of grafts. In renal transplantation, the technique could contribute to reduce the incidence of delayed graft function. In addition, the procedure is compatible with surgical rapid recovery in hypothermia when there is also lung donation. The future lies in the consolidation of cardiac donation by extending normothermic perfusion to the thoracic cavity.

Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models.

Polycystic kidney disease is a complex clinical entity which comprises a group of genetic diseases that leads to renal cyst development. We evaluated the most suitable housekeeping genes for microRNA expression by RT-qPCR analyses of kidney tissues in Pkd1-deficient mouse models from a panel of five candidates genes (miR-20a, miR-25, miR-26a, miR-191 and U6) and 3 target genes (miR-17, miR-21 and let-7a) using samples from kidneys of cystic mice (Pkd1flox/flox:Nestincre, CY), non-cystic controls (Pkd1flox/flox, NC), Pkd1-haploinsufficient (Pkd1+/-, HT), wild-type controls (Pkd1+/+, WT), severely cystic mice (Pkd1V/V, SC), wild-type controls (CO). The stability of the candidate genes was investigated using NormFinder, GeNorm, BestKeeper, DataAssist, and RefFinder software packages and the comparative ΔCt method. The analyses identified miR-26a as the most stable housekeeping gene for all kidney samples, miR-20a for CY and NC, miR-20a and miR-26a for HT and WT, and miR-25 and miR-26a for SC and CO. Expression of miR-21 was upregulated in SC compared to CO and trends of miR-21 upregulation and let-7a downregulation in CY and HT compared to its control kidneys, when normalized by different combinations of miR-20a, miR-25 and miR-26a. Our findings established miR-20a, miR-25, and miR-26a as the best housekeeping genes for miRNA expression analyses by RT-qPCR in kidney tissues of Pkd1-deficient mouse models.

The inhibition of neutrophil granule enzyme secretion and chemotaxis by pertussis toxin.

Pertussis toxin treatment of rabbit peritoneal neutrophils causes a concentration-dependent inhibition of granule enzyme secretion induced by formylmethionyl-leucyl-phenylalanine, C5a, and leukotriene B4. It also inhibits chemotaxis induced by formylmethionyl-leucyl-phenylalanine. The same toxin treatment, however, has no effect on granule enzyme secretion induced by the calcium ionophore A23187 or phorbol 12-myristate 13-acetate. Moreover, pertussis toxin treatment does not affect either the number or affinity of the formylpeptide receptors on the neutrophil nor does it have any effect on the unstimulated levels of cyclic AMP (cAMP) or the transient rise in cAMP induced by chemotactic factor stimulation in these cells. We hypothesize that pertussis toxin, as in other cells, interacts with a GTP binding regulatory protein identical with or analogous to either Ni or transducin which mediates the receptor-induced inhibition or activation of a target protein or proteins required in neutrophil activation. The nature of the target protein is unknown, but it is not the catalytic unit of adenylate cyclase. The target protein acts after binding of chemotactic factor to its receptor in the sequence that leads to the receptor-induced rise in intracellular Ca2+. It does not affect the responses elicited by the direct introduction of calcium into the cells or the activity of protein kinase C.

ADP-ribosyltransferase activity of pertussis toxin and immunomodulation by Bordetella pertussis.

Pertussis toxin is produced by the causative agent of whooping cough, Bordetella pertussis, and is an adenosine diphosphate (ADP)-ribosyltransferase capable of covalently modifying and thereby inactivating many eukaryotic G proteins involved in cellular metabolism. The toxin is a principal determinant of virulence in whooping cough and is a primary candidate for an acellular pertussis vaccine, yet it is unclear whether the ADP-ribosyltransferase activity is required for both pathogenic and immunoprotective activities. A B. pertussis strain that produced an assembled pertussis holotoxin with only 1 percent of the ADP-ribosyltransferase activity of the native toxin was constructed and was found to be deficient in pathogenic activities associated with B. pertussis including induction of leukocytosis, potentiation of anaphylaxis, and stimulation of histamine sensitivity. Moreover, this mutant strain failed to function as an adjuvant and was less effective in protecting mice from intracerebral challenge infection. These data suggest that the ADP-ribosyltransferase activity is necessary for both pathogenicity and optimum immunoprotection. These findings bear directly on the design of a nontoxic pertussis vaccine.

The use of ECMO in ICU. Recommendations of the Spanish Society of Critical Care Medicine and Coronary Units. / Empleo de ECMO en UCI. Recomendaciones de la Sociedad Española de Medicina Intensiva Crítica y Unidades Coronarias.

The use of extracorporeal membrane oxygenation systems has increased significantly in recent years; given this reality, the Spanish Society of Critical Intensive Care Medicine and Coronary Units (SEMICYUC) has decided to draw up a series of recommendations that serve as a framework for the use of this technique in intensive care units. The three most frequent areas of extracorporeal membrane oxygenation systems use in our setting are: as a cardiocirculatory support, as a respiratory support and for the maintenance of the abdominal organs in donors. The SEMICYUC appointed a series of experts belonging to the three working groups involved (Cardiological Intensive Care and CPR, Acute Respiratory Failure and Transplant work group) that, after reviewing the existing literature until March 2018, developed a series of recommendations. These recommendations were posted on the SEMICYUC website to receive suggestions from the intensivists and finally approved by the Scientific Committee of the Society. The recommendations, based on current knowledge, are about which patients may be candidates for the technique, when to start it and the necessary infrastructure conditions of the hospital centers or, the conditions for transfer to centers with experience. Although from a physiopathological point of view, there are clear arguments for the use of extracorporeal membrane oxygenation systems, the current scientific evidence is weak, so studies are needed that define more precisely which patients benefit most from the technique and when they should start.

[Association between the frequency and duration, but not the intensity, of headache with mechanical hypersensitivity and the health of patients with tension-type headache]. / Asociacion de la frecuencia y duracion, pero no la intensidad, de la cefalea con la hipersensibilidad mecanica y la salud en pacientes con cefalea de tension.

INTRODUCTION: The association between headache clinical parameters and other health-related and neuro-physiological outcomes is controversial. AIM: To investigate the association between headache frequency, duration and intensity with cranio-cervical pressure sensitivity considering the interaction of health-related and physical outcomes. PATIENTS AND METHODS: Seventy-two individuals with tension type headache were included. A 1-month diary was used to assess headache frequency, duration and intensity. Pressure pain thresholds (PPT) and peri-cranial tenderness to palpation, health-related quality of life (Short Form-36), disability, depression, and cervical range of motion were the outcomes. All outcomes were introduced into hierarchic logistic regression models to assess potential associations. RESULTS: Several associations between headache frequency and duration, but not intensity, with the remaining variables were found. Regression analysis showed that PPT over the temporalis muscle, bodily pain, age and physical role explained the 22.3% of the headache frequency, whereas general health, PPT over the upper trapezius and headache frequency explained 20% of headache duration (p < 0.001). CONCLUSIONS: This study found that headache frequency and duration, but not headache intensity, were associated with neurophysiological outcomes, e.g., cranio-cervical pressure sensitivity, and bodily pain in tension type headache. Other variables including age, physical role and general health were also associated with headache frequency and duration.

TITLE: Asociacion de la frecuencia y duracion, pero no la intensidad, de la cefalea con la hipersensibilidad mecanica y la salud en pacientes con cefalea de tension.Introduccion. La asociacion entre las variables clinicas de la cefalea y otras variables neurofisiologicas o de salud es controvertida. Objetivo. Determinar la asociacion de la frecuencia, duracion e intensidad del dolor con la sensibilidad mecanica craneocervical, considerando el efecto de otras variables de salud y fisicas. Pacientes y metodos. Se incluyeron 72 pacientes con cefalea de tension. Se mantuvo un diario mensual para determinar la frecuencia, duracion e intensidad del dolor. Se calcularon los umbrales de dolor a la presion (UDP) y la sensibilidad a la palpacion sobre la region craneocervical, calidad de vida (Short Form-36), discapacidad, depresion y movilidad cervical. Se introdujeron todas las variables en modelos de regresion logistica jerarquica para determinar las interacciones. Resultados. Se encontraron correlaciones entre la frecuencia y duracion de la cefalea, pero no la intensidad, con el resto de variables. El analisis de regresion mostro que la combinacion del UDP sobre el temporal, dolor corporal, edad y rol fisico explicaba el 22,3% de la varianza de la frecuencia, mientras que la combinacion de salud general, UDP sobre el trapecio superior y frecuencia del dolor explicaba el 20% de la varianza de la duracion (p < 0,001). Conclusiones. Este estudio ha encontrado que la frecuencia y la duracion de la cefalea, pero no la intensidad, se encuentran asociadas con variables neurofisiologicas, como el UDP sobre el cuello/cabeza o la percepcion de dolor generalizado en la cefalea tensional. Otras variables, como la edad, el rol fisico y la salud general tambien mostraron una asociacion con la frecuencia y la duracion de la cefalea.

Polymorphic oxidation of debrisoquine in lung cancer patients.

Oxidative polymorphism of debrisoquine (DBQ) was assessed in 84 patients (81 male) with histologically proven bronchogenic carcinoma and in 143 healthy male smokers. 80 (95%) patients and 133 (93%) controls, with a metabolic ratio (MR) below 12.6, were classified as extensive metabolisers of DBQ (no significant difference between patients and controls). Only 1 of the 73 patients with epidermoid or microcytic carcinomas was classified as a poor metaboliser (PM) (P = 0.031 compared with controls). 63 patients (75%) and 110 controls (77%) showed a very fast oxidative rate, with MR values under 1 (not significant). The EM phenotype of DBQ might be a secondary genetic risk factor for developing bronchogenic carcinoma in male smokers.

Production of experimental allergic encephalomyelitis with the aid of pertussigen in mouse strains considered genetically resistant.

Strains of mice (BALB/c An Bradley/Wehi, C57B1/10J, CBA/ca H Wehi, DBA/2 Wf, A/J Wehi), thought to be genetically resistant to experimental allergic encephalomyelitis (EAE) and known to be resistant to becoming hypersensitive to histamine after administration of pertussis vaccine were tested for their ability to develop EAE when purified pertussigen was included in the immunization. It was found that C57B1/10J, CBA/ca H Wehi, BALB/c An Bradley/Wehi and DBA/2 Wf developed typical signs and histologic evidence of EAE. The A/J Wehi and the B10D2/n Sn (not previously tested) strains developed only mild signs of EAE, while the known susceptible (SJL/J X BALB/c An Bradley/Wehi) F1 hybrids developed severe EAE. Histologic evidence of EAE was lacking in A/J Wehi mice and was minimal in B10D2/n Sn mice. These results suggest that neither the H-2 complex nor the gene controlling susceptibility to sensitization to histamine by administration of pertussigen are wholly responsible for susceptibility to EAE.

CYP2D6 genotypes in Spanish women with breast cancer.

Wild type and three mutated alleles of the polymorphic CYP2D6 gene were studied in genomic DNA samples from 187 women with breast carcinoma and 151 healthy women by a mutation-specific polymerase chain reaction. The prevalence of the enzyme-inactivating CYP2D6(B) allele was higher among patients (18.2%) than in controls (11.6%; OR = 1.7; 95% c.i. = 1.14-3.13; P = 0.018). This excess was more marked in postmenopausal patients (19.8%, P = 0.0086) and in patients with non-ductal infiltrating carcinomas (25.8%, P = 0.003). The percentage of carriers of only one active gene (heterozygote extensive metabolizers) was higher in patients (31% vs. 19.9%; OR = 1.81; 95% c.i. = 1.06-3.11; P = 0.02). The CYP2D6(B)-carrier state may be related to a greater risk of breast cancer in women.

Early and late exercise testing. Usefulness after percutaneous transluminal coronary angioplasty.

Seventy-five patients 36 to 68 years of age were studied after undergoing successful single-vessel percutaneous transluminal coronary angioplasty (PTCA). In all cases exercise tests (ETs) were made prior to the procedure and 2 to 4 days and 6 months thereafter. Angiographic controls were performed on patients with positive ET. Both early and late ET carried out after dilatation lasted significantly longer. Patients were able to tolerate greater work loads and their heart rates and arterial blood pressures also were higher. The ETs conducted prior to PTCA were positive in 62 patients. After dilatation, the early test was positive in 18 and the late test, in 10. Angiographic controls revealed restenosis in 75 percent of patients with single-vessel disease and positive ET. Thus, the results of ETs in patients with single-vessel disease tend to become negative after successful PTCA. Early and late positive ETs are associated with restenosis, while negative ETs indicate a good prognosis.

Role of glucocorticoids in early T-cell differentiation.

The results of the T-cell differentiation in the progeny of adrenalectomized pregnant rats (Adx fetuses), an experimental model that ensures the absence of glucocorticoids (GCs) during the first stages of development, are summarized. In Adx thymuses there is an accelerated maturation of thymocytes that is reversed by in vivo GC replacement. In addition, Adx thymuses show decreased cell content, which correlates with both the increased numbers of apoptotic cells and an early migration of DP (CD4+CD8+) and SP (both CD4+CD8- and CD4-CD8+) thymocytes to the spleen. As shown by in vitro recolonization assays, accelerated T-cell differentiation is a consequence of changes in the biology of lymphoid precursors occurring in the fetal liver of Adx fetuses. They arrive at the thymic primordium earlier and mature faster than the fetal liver lymphoid progenitors from Sham control fetuses. After the establishment of a fetal hypothalamus-pituitary-gland-adrenal-gland (HPA) axis, there is a gradual normalization of the T-cell development Adx fetuses.

Reexploration for hemorrhage following coronary artery bypass grafting: incidence and risk factors. Northern New England Cardiovascular Disease Study Group.

OBJECTIVE: To assess mortality and risk factors associated with reexploration for hemorrhage in patients undergoing coronary artery bypass grafting (CABG). DESIGN: Regional cohort study. Patient characteristics, treatment variables, and outcome measures were collected prospectively. SETTING: All 5 centers performing cardiac surgery in Maine, New Hampshire, and Vermont. PATIENTS: A consecutive cohort of 8586 patients undergoing isolated CABG between 1992 and 1995. MAIN OUTCOME MEASURES: Postoperative hemorrhage leading to reexploration, in-hospital mortality, and length of stay. RESULTS: A total of 305 patients (3.6%) underwent reexploration for bleeding. In these patients, in-hospital mortality was nearly 3 times higher (9.5% vs 3.3% for patients not requiring reoperation, P<.001) and average length of stay from surgery to discharge was significantly longer (14.5 days vs 8.6 days, P<.001). High rates of reexploration for hemorrhage were observed in patients with prolonged (> 150 minutes) cardiopulmonary bypass (39 [11.1%] of 351) and in those requiring an intra-aortic balloon pump intraoperatively (12 [8%] of 139). In multivariate analysis, older age, smaller body surface area, prolonged cardiopulmonary bypass, and number of distal anastomoses were associated with increased bleeding risks. The use of thrombolytic therapy within 48 hours of surgery was weakly but not significantly associated with the need for reexploration. Factors not significantly associated with reexploration included patient sex, preoperative ejection fraction, surgical priority, history of liver disease, myocardial infarction, prior CABG, renal failure, and diabetes mellitus. CONCLUSIONS: Hemorrhage requiring reexploration after CABG is associated with markedly increased mortality and length of stay. Patients predicted to have increased risks of bleeding may benefit from prophylactic use of aprotinin, aminocaproic acid, or other agents shown to reduce hemorrhage.

Effect of preoperative aspirin use on mortality in coronary artery bypass grafting patients.

BACKGROUND: Discontinuing aspirin use in patients before coronary artery bypass grafting (CABG) has focused on bleeding risks. The effect of aspirin use on overall mortality with this procedure has not been studied. METHODS: We performed a case patient-control patient study of the 8,641 consecutive isolated CABG procedures performed between July 1987 and May 1991 in Maine, New Hampshire, and Vermont. Patients included all 368 deaths. Each case patient was paired with approximately two matched survivors (control patients). Aspirin use was defined by identification of ingestion within 7 days before the operation. RESULTS: CABG patients using preoperative aspirin were less likely to experience in-hospital mortality in univariate (odds ratio [OR] = 0.73, 95% confidence interval [0.54, 0.97]) and multivariate [OR = 0.55, (0.31, 0.98)] analysis compared to nonusers. No significant difference was seen in the amount of chest tube drainage, transfusion of blood products, or need for reexploration for hemorrhage between patients who did and did not receive aspirin. CONCLUSIONS: Preoperative aspirin use appears to be associated with a decreased risk of mortality in CABG patients without significant increase in hemorrhage, blood product requirements, or related morbidities.