Coenzyme Q10 supplementation during in vitro maturation of bovine oocytes (Bos taurus) helps to preserve oocyte integrity after vitrification.

Oocyte vitrification causes less cell stress than slow cooling, but cytoskeletal and spindle alterations may occur affecting the oocyte competence. In vitro maturation (IVM) supplementation with different antioxidant molecules has been performed to attenuate this harmful stress. Coenzyme Q10 (CoQ10 ) supplementation has previously shown to have positive effects in bovine and mouse in vitro embryo development. The aim of this study was to evaluate the effects of CoQ10 during bovine oocyte IVM and vitrification. Cumulus-oocyte complexes (COCs) (n = 311) were cultured under standard maturation conditions with 0 µM (control), 25 µM and 50 µM CoQ10 supplementation. After 22 hr, a cohort of 170 oocytes both from the control and from CoQ10 -supplemented groups were vitrified, warmed and returned to incubation until 24 hr of maturation, while the rest of the oocytes (n = 141) remained fresh. Then, oocyte survival was assessed morphologically by stereomicroscopy. Oocytes from all groups were then fixed and stained for assessing cortical granules (CG) migration and nuclear stage. High rates of oocyte MII progression and appropriate CG migration as a continuous layer beneath the plasma membrane were obtained both in control and in CoQ10 groups. Results showed that although vitrification has great impact in survival of IVM bovine oocytes, 50 µM CoQ10 supplementation significantly improved oocyte survival (p = .045) and reduced the premature CG exocytosis, helping to preserve the CG migration pattern (31.3% control vs. 54.5% in 50 µM CoQ10 ; p = .039), attenuating the negative effects of vitrification.

Prevalence and factors associated with nontuberculous mycobacteria in non-cystic fibrosis bronchiectasis: a multicenter observational study.

BACKGROUND: Data on the prevalence of and factors associated with nontuberculous mycobacteria (NTM) in patients with non-cystic fibrosis (CF) bronchiectasis are limited. Our aim was to determine the prevalence and factors associated with isolation of NTM in this population. METHODS: We performed a multicenter observational study of historical cohorts comprising consecutive patients with non-CF bronchiectasis and at least 2 sputum samples cultured for mycobacteria over a period of 5 years. RESULTS: The study population included 218 adult patients (61.9 % women) with a mean (SD) age of 55.7 (16) years and a mean (SD) of 5.1 (3.3) cultures/patient. NTM was isolated from sputum in 18 patients (8.3 %). Of these, 5 patients (28 %) met the American Thoracic Society criteria for NTM disease. Mycobacterium avium complex was the most frequently isolated microorganism (9 patients, 4.1 %). The variables independently associated with isolation of NTM were FVC ≥ 75 % predicted (OR, 4.84; 95 % CI 1.47 to 15.9; p < 0.05), age ≥ 50 years (OR, 4.74; 95 % CI 1.25 to 17.97; p < 0.05), and body mass index (BMI) ≤ 23 kg/m(2) (OR, 2.97; 95 % CI 1.03-8.58; p < 0.05). Patients with these three characteristics had a 40 % probability of having at least one isolation of NMT. CONCLUSIONS: A significant number of patients with non-CF bronchiectasis are positive for the isolation of NTM. M. avium complex is the most frequently isolated mycobacteria. FVC ≥ 75 % predicted, age ≥ 50 years, and a BMI ≤ 23 kg/m(2) were independently associated with the presence of NTM in patients with non-CF bronchiectasis.

Coagulation profiles of unexpected DCDD donors do not indicate a role for exogenous fibrinolysis.

It has been suggested that vascular stasis during cardio-circulatory arrest leads to the formation of microvascular thrombi and the viability of organs arising from donation after circulatory determination of death (DCDD) donors may be improved through the application of fibrinolytic therapy. Our aim was to comprehensively study the coagulation profiles of Maastricht category II DCDD donors in order to determine the presence of coagulation abnormalities that could benefit from fibrinolytic therapy. Whole blood from potential DCDD donors suffering out-of-hospital cardiac arrest was sampled after declaration of death in the emergency department, and rotational thromboelastomeric analysis was performed. Between July 2012 and December 2013, samples from 33 potential DCDD donors were analyzed. All patients demonstrated hyperfibrinolysis (HF), as reflected by maximum clot lysis of 98-100% in all cases, indicating that there is no role for additional fibrinolytic therapy in this setting. As well, we observed correlations between thromboelastomeric lysis parameters and maximum hepatic transaminase levels measured in potential donors and renal artery flows measured during ex situ hypothermic oxygenated machine perfusion, indicating that further studies on the utility of thromboelastometry to evaluate organ injury and perhaps even viability in unexpected DCDD may be warranted.

Gender perspective in the implementation of hygiene in cardiology intensive care.

INTRODUCTION: Hygiene in critical patients is an essential daily care, provided under safe conditions, to promote comfort and maintain the integrity of skin and mucous membranes, however, it can generate feelings of dependence and vulnerability in patients. The aim of this post hoc study is to know the differences in satisfaction and lived experience regarding bed hygiene in an intensive care unit according to biological sex and gender perspective. METHODS: Observational, descriptive and prospective study in which an ad hoc questionnaire was administered to 148 conscious and oriented patients of legal age. The questionnaire was completed 24-48 hours after admission to the unit, once the initial bed hygiene had been performed. RESULTS: Males experienced conformism (51%), embarrassment (31%) and relief (9%); Women felt conformism (35.4%), embarrassment (18.8%) and relief (29.2%) (p < 0.05). Women experienced a feeling of cleanliness in 89.1% compared to 56.1% of men (p < 0.05). Men were offered to wash their genitals in 72.9% compared to 35.7% of women (p < 0.05). 34.3% of men would prefer a family member to assist them during hygiene (62.9% by their wives), compared to 27.1% of women who would prefer a family member (84.6% by their daughters). CONCLUSION: Women tolerate bed hygiene better than men and appreciate more the feeling of cleanliness. Women are identified as caregivers, both professionally and in the family, and patients prefer them to collaborate in the performance of hygiene, being wives preferred by men and daughters preferred by women.

Mitral valve-in-valve implantation: A case report.

INTRODUCTION: Mitral valve-in-valve implantation is a new therapeutic tool in the field of structural interventional cardiology for patients with bioprosthetic dysfunction due to severe mitral valve regurgitation and high surgical risk. The objective was to develop an individualised nursing care plan for a patient undergoing this procedure; the first case in our centre. CASE DESCRIPTION: A 75-year-old woman, independent for activities of daily living, with a history of chronic renal failure and biological mitral valve replacement due to rheumatic valve disease. She was admitted to the acute cardiac care unit for severe symptomatic mitral valve regurgitations secondary to mitral bioprosthesis dysfunction. Heart surgery was ruled out due to comorbidities and high surgical risk, and the patient underwent percutaneous mitral valve-in-valve implantation. The implantation was successful. ASSESSMENT: The nursing assessment followed Marjory Gordon's conceptual model identifying the following impaired patterns: pattern 2: bilateral malleolar oedema without pitting; pattern 3: urinary catheter and intravenous diuretic use; pattern 4: dyspnoea on moderate exertion, dry nocturnal cough, orthopnoea and respiratory disturbances, and activity intolerance; pattern 5: need for pharmacological assistance for a good night's rest. DIAGNOSES: The following nursing diagnoses were established using the NANDA taxonomy: Excess fluid volume; ineffective breathing pattern; Activity intolerance and problem collaborating: Hypotension and anaemia secondary to deep thigh haematoma. PLANNING: The following objectives were set based on the NOC taxonomy: Fluid balance; Respiratory status: ventilation; Cardiopulmonary status and the following NIC interventions: Hypervolaemia management; Respiratory monitoring and oxygen therapy; Vital sign monitoring and heart care. DISCUSSION: Nursing interventions aimed at monitoring haemodynamic status, fluid restriction together with the efficacy of diuretic treatment achieved a negative water balance which contributed to fluid depletion improving respiratory symptoms, enabling implantation under better conditions. CONCLUSIONS: Technological progress in the health sciences, and in the field of acute cardiology in particular, directly calls for training, revision and updating of critical care nursing. Given this dynamic and continually evolving process, the specialist intensive care nurse, the inclusion of the cardiovascular nurse specialist in multidisciplinary teams such as the heart team, and expanding the consultation of the haemodynamic nurse are urgently required to ensure optimal nursing care, safety, and care quality.

[Cerebrovascular accident with haemorrhagic transformation in a patient on antiplatelet treatment subjected to surgery of a hypophyseal macroadenoma]. / Accidente cerebrovascular con transformación hemorrágica en una paciente con tratamiento antiagregante intervenida de macroadenoma hipofisario.

The case is presented of a 51 year-old woman, proposed for endoscopic endonasal transsphenoidal resection of a hypophyseal macroadenoma diagnosed in the context of a stroke suffered 10 weeks before the date of the surgery. During this time, she had been treated with antiplatelet drugs, which were withdrawn 5 days before the surgery. The surgical procedure was performed without any incidents. On the second day after the surgery, the patient had an ischaemic infarction of the left cerebellar hemisphere, with signs of hydrocephaly and a posterior haemorrhagic transformation, with brain death 5 days after the operation. There are no definitive guidelines on the use of antiplatelet drugs in the perioperative period of neurosurgery. Also, there is no agreement as regards the waiting time between a cerebrovascular event and surgery, it appears that between 4 and 12 weeks would be the most advisable. The importance of an individual assessment of each patient before surgery is emphasised, as well as a review of the antiplatelet management of the patient with a risk of thrombosis in the context of neurosurgery, and their possible postoperative complications.

Regulation of c-fos expression in transgenic mice requires multiple interdependent transcription control elements.

Transcription control regions of eukaryotic genes contain multiple sequence elements proposed to function independently to regulate transcription. We developed transgenic mice carrying fos-lacZ fusion genes with clustered point mutations in each of several distinct regulatory sequences: the sis-inducible element, the serum response element, the fos AP-1 site, and the calcium/cAMP response element. Analysis of Fos-lacZ expression in the CNS and in cultured cells demonstrated that all of the regulatory elements tested were required in concert for tissue- and stimulus-specific regulation of the c-fos promoter. This implies that the regulation of c-fos expression requires the concerted action of multiple control elements that direct the assembly of an interdependent transcription complex.

Predicting high risk of exacerbations in bronchiectasis: the E-FACED score.

BACKGROUND: Although the FACED score has demonstrated a great prognostic capacity in bronchiectasis, it does not include the number or severity of exacerbations as a separate variable, which is important in the natural history of these patients. OBJECTIVE: Construction and external validation of a new index, the E-FACED, to evaluate the predictive capacity of exacerbations and mortality. METHODS: The new score was constructed on the basis of the complete cohort for the construction of the original FACED score, while the external validation was undertaken with six cohorts from three countries (Brazil, Argentina, and Chile). The main outcome was the number of annual exacerbations/hospitalizations, with all-cause and respiratory-related deaths as the secondary outcomes. A statistical evaluation comprised the relative weight and ideal cut-off point for the number or severity of the exacerbations and was incorporated into the FACED score (E-FACED). The results obtained after the application of FACED and E-FACED were compared in both the cohorts. RESULTS: A total of 1,470 patients with bronchiectasis (819 from the construction cohorts and 651 from the external validation cohorts) were followed up for 5 years after diagnosis. The best cut-off point was at least two exacerbations in the previous year (two additional points), meaning that the E-FACED has nine points of growing severity. E-FACED presented an excellent prognostic capacity for exacerbations (areas under the receiver operating characteristic curve: 0.82 for at least two exacerbations in 1 year and 0.87 for at least one hospitalization in 1 year) that was statistically better than that of the FACED score (0.72 and 0.78, P<0.05, respectively). The predictive capacities for all-cause and respiratory mortality were 0.87 and 0.86, respectively, with both being similar to those of the FACED. CONCLUSION: E-FACED score significantly increases the FACED capacity to predict future yearly exacerbations while maintaining the score's simplicity and prognostic capacity for death.

Magnesium sulphate and (123)I-MIBG in pheochromocytoma: Two useful techniques for a complicated disease.

Pheochromocytoma is a tumour of the chromaffin tissue. It may, through catecholamine release, have deleterious effects on myocardial structure. A 48-year-old woman with a history of hypertension and type II diabetes mellitus (ASA II) was diagnosed of pheochromocytoma-induced myocarditis, which caused severe cardiogenic shock, with an ejection fraction of 20%. Extreme blood pressure swings required aggressive therapy with vasoactive drugs (norepinephrine and dopamine) and an intra-aortic balloon pump, despite which severe haemodynamic instability persisted. Finally, the use of magnesium sulphate allowed for cardiovascular stabilization and weaning off vasoactive drugs prior to surgery. (123)I-metaiodobenzylguanidine scintigraphy helps not only to functionally confirm tumour tissue, but also to assess severity and prognosis of cardiac failure. Prognosis of pheochromocytoma-induced heart failure can be very poor. The use of these two well-known and relatively simple 'tools' for treatment and prognosis is a helpful option to keep in mind.

Identification of 26 new constitutional RB1 gene mutations in Spanish, Colombian, and Cuban retinoblastoma patients.

Constitutional mutations in the RB1 gene predispose to retinoblastoma development. Hence genetic screening of retinoblastoma patients and relatives is important for genetic counseling purposes. In addition, RB1 gene mutation studies may help decipher the molecular mechanisms leading to tumors with different degrees of penetrance or expressivity. In the course of genetically screening of 107 hereditary and non-hereditary retinoblastoma patients (11 familiar bilateral, 4 familiar unilateral, 49 sporadic bilateral and 43 sporadic unilateral) and kindred from Spain, Colombia and Cuba, using direct PCR sequencing, we observed 45 distinct mutations and four RB1 deletions in 53 patients (9 familiar bilateral, 2 familiar unilateral, 31 sporadic bilateral and 11 sporadic unilateral). Most of these mutations (26/45, 57%) have not been reported before. In 32 patients, the predisposing mutations correspond to nonsense (mainly CpG transitions) and small insertions or deletions whose expected outcome is a truncated Rb protein that lacks the functional pockets and tail. Five single aminoacid replacements and seventeen mutations affecting splicing sites were also observed in retinoblastoma patients. Two of these sixteen mutations are of unclear pathogenic nature.

Spectrum of germline RB1 gene mutations in Spanish retinoblastoma patients: Phenotypic and molecular epidemiological implications.

Mutation analysis of retinoblastoma is considered important for genetic counseling purposes, as well as for understanding the molecular mechanisms leading to tumors with different degrees of penetrance or expressivity. In the course of an analysis of 43 hereditary retinoblastoma Spanish patients and kindred, using direct PCR sequencing, we have observed 29 mutations; most of them (62%) have not been reported previously. Of the mutations, 69% correspond to nonsense mutations (mainly CpG transitions) and frameshifts, with the expected outcome of a truncated Rb protein that lacks the functional pocket domains and tail. The remainder corresponds to splicing mutations, most of them (62%) targeted to invariant nucleotides, with the predicted consequence of out of frame exon skipping. Two of the splicing mutations in our study were found associated to families with a low-penetrance phenotype. Additionally, most of the mutations affecting splice junctions corresponded to retinoblastoma cases of either sporadic or hereditary nature with delayed onset (32 months on average). In contrast, most of the nonsense and frameshift mutations are associated with an early age at diagnosis (8.7 months on average). These differences are discussed in the context of the relationships between genotype and low expressivity phenotype. The differences in the spectrum of RB1 mutations found in this and other European surveys are also discussed in the context of alternate DNA methylation and mismatch repair phenotypes.

A gene expression approach to mapping the functional maturation of the hippocampus.

Previous studies have shown an association among seizures, neuronal death and the expression of cellular immediate-early genes (cIEG). To understand further the relationship between these processes, we investigated the ability of kainic acid (KAI) to induce behavioral responses and gene expression in the hippocampus of developing fos-lacZ transgenic mice. Despite the fact that KAI elicited seizure-like activity from P2 onwards, Fos-lacZ was first detected at P5 in CA3 pyramidal neurons. Thus, intense behavioral responses were not invariably associated with fos-lacZ expression. Furthermore, while adult CA3 neurons are highly susceptible to KAI toxicity, they are resistant at P5. Therefore, the presence of Fos-lacZ in CA3 neurons is not necessarily predictive of their fate. By P10, Fos-lacZ was induced in CA3 neurons and in the most mature granule neurons of the dentate gyrus (DG). Between P15 and P20, KAI induced fos-lacZ in all CA1 and CA3 pyramidal neurons and most granule neurons of the DG. This stereotypical pattern of fos-lacZ expression mirrors the ontogeny of hippocampal circuitry and glutamate signalling. Thus the fos-lacZ mice can be used to map the functional maturation of the nervous system with single cell resolution. The scope of this approach was extended by administration of additional chemoconvulsants to fos-lacZ mice and by analysis of fos-lacZ transgenic mice with mutations in their FAP site. These additional studies revealed anatomical and mechanistic differences in glutamate receptor-mediated transcriptional responses in the nervous system.

Effect of different convulsants on calmodulin levels and proto-oncogene c-fos expression in the central nervous system.

In the present study, a relationship between convulsant activity and two cellular events, changes in calmodulin (CaM) concentration and proto-oncogene c-fos expression has been considered. c-fos has been found activated after the administration of the organochlorine insecticide lindane, the Ca2+ channel agonist Bay K, and N-methyl-D-aspartate (NMDA). The administration of the voltage-dependent Ca2+ channel antagonist nifedipine was able to block the expression elicited by lindane. The effect of lindane on c-fos expression could not be blocked by prior administration of MK-801, a non-competitive antagonist of the NMDA receptor. These results suggest a possible role for the voltage-dependent Ca2+ channels in the mechanism of action of lindane. By means of in situ hybridization, the different patterns of c-fos expression after the administration of the mentioned compounds have been described. A possible modification of the levels of CaM has also been investigated. Among all the subcellular fractions considered, only levels of nuclear CaM appeared to be affected after the different treatments. The changes observed seemed to follow a similar pattern to that described for c-fos induction. Calcium entry through these voltage-dependent calcium channels would be the link between membrane depolarizing events and expression of c-fos and/or increase in nuclear CaM.

Empiric treatments impair the diagnostic yield of BAL in HIV-positive patients.

OBJECTIVE: To investigate the influence of empiric treatments prior to fiberoptic bronchoscopy (FOB) on the diagnostic yield of BAL in HIV-positive patients with respiratory symptoms. METHOD: We studied 123 consecutive FOBs with BAL in HIV-positive patients; 101 of these patients (82%) had received previous antimicrobial treatment from 1 to 60 days. Diagnostic yield of BAL for Pneumocystis carinii, Mycobacterium tuberculosis, and bacterial pneumonia was compared between patients with and without previous empiric treatments. RESULTS: A diagnosis was obtained in 85 patients (69%), of whom 17 (20%) had multiple infections. Diagnostic yield was higher in patients without previous treatment, 91% (20/22) compared with 64% (65/101), p < 0.03. Diagnostic yield was also higher for bacterial pneumonia: seven isolations from 22 patients not receiving previous empiric treatment (32%), compared with 11 of those who had (11%; p < 0.02). The duration of empiric treatment against P carinii in patients in whom it was isolated was significantly shorter than in those in whom P carinii was not detected (3.5 +/- 1.8 days compared with 5.2 +/- 2.4 days; p = 0.003). FOB permitted a change in treatment in 62% of patients with a final diagnosis. CONCLUSION: This study demonstrates that empiric treatments prior to FOB significantly impair the diagnostic yield of BAL in detecting common pathogens in HIV-infected patients with respiratory symptoms.

Specific antibody response against the 23-valent pneumococcal vaccine in patients with alpha(1)-antitrypsin deficiency with and without bronchiectasis.

OBJECTIVE: To assess the specific antibody response against polyvalent pneumococcal vaccine in patients with alpha(1)-antitrypsin deficiency (AATD) and respiratory infections. DESIGN AND PARTICIPANTS: We investigated specific IgG, IgG1, and IgG2 antibody responses against the 23-valent antipneumococcal vaccine in 18 patients with AATD phenotype PiZZ, 9 of whom had bronchiectasis and 4 a history of recurrent pneumonia, and compared them with a control group of 40 healthy volunteers. INTERVENTIONS: Blood samples were drawn just prior to and 3 weeks after immunization. MEASUREMENTS AND RESULTS: Quantification of specific IgG and its subclasses was performed by an enzyme-linked immunosorbent assay. For patients with AATD, mean increases in specific antipneumococcal titers were 4.7-fold (25 to 75% quartiles, 2.5- to 6.8-fold) for total IgG, 3.2-fold (1.2- to 4.9-fold) for IgG1, and 2.1-fold (1.8- to 3.7-fold) for IgG2. For the control group, the values were 3.3-fold (1.8- to 5.8-fold) for total IgG, 2. 5-fold (1.9- to 3.4-fold) for IgG1, and 3.1-fold (1.9- to 4.5-fold) for IgG2; differences were not significant. Patients with bronchiectasis showed a tendency toward higher levels of IgG subclasses than both control subjects and patients without bronchiectasis; however, there was a tendency toward lower postvaccination serum levels of specific antipneumococcal IgG, IgG1, and IgG2 in patients with bronchiectasis compared with patients without bronchiectasis, but this trend did not reach statistical significance. Three of the four patients with recurrent pneumonia did not show an appropriate IgG2 response. CONCLUSIONS: These results suggest that, as a group, patients with AATD have a preserved antibody response against pneumococcal polysaccharides. Patients with bronchiectasis show a tendency toward a decreased antibody response, even with increased serum levels of most Ig types. Individuals with an impaired IgG2 response seem to be at increased risk of recurrent pneumonia. Considering the pernicious effect of pulmonary infections on these patients and the preserved antibody response in a majority of them, pneumococcal vaccination should be recommended to patients with AATD.

Lindane-induced convulsions in NMRI and OF1 mice: antagonism with (+)MK-801 and voltage-dependent calcium channel blockers.

The convulsant profile of lindane was investigated in OF1 and NMRI mice lines in relation to other convulsants acting at the GABAA and NMDA receptor complexes. Thus, a specific GABA-gated chloride channel blocker, PTX, a GABAA receptor antagonist, PTZ, and an excitatory amino acid receptor agonist, NMDA, were used. Antagonism of the convulsant effects of each of these drugs was investigated with (+)MK-801, a blocker of the NMDA-operated cation channel, and with nifedipine, a voltage-dependent calcium channel antagonist. While no differences in potency for PTX or PTZ to induce seizures were observed between OF1 and NMRI mice, lindane was approximately 80 and 90% more potent in its ability to induce seizures and lethality, respectively, in OF1 than in NMRI mice. Brain lindane concentrations at the moment of convulsion, measured after ED100 doses of lindane (400 and 200 mg/kg for NMRI and OF1 mice, respectively), did not differ between OF1 and NMRI mice, suggesting that the different potency of lindane between these mouse lines is a consequence of pharmacokinetic factors. Furthermore, (+)MK-801 antagonized seizures induced by either lindane, PTX or PTZ with similar potencies in both mouse lines. These results, coupled with the different pharmacokinetics of lindane in OF1 and NMRI mice, suggest that the distinct effects of lindane in these mice are not mediated by different activities at either NMDA or GABAA receptor complexes. Nonetheless, nifedipine antagonized lindane-induced seizures with a three-fold higher potency in NMRI than in OF1 mice. In contrast, nifedipine failed to antagonize PTX and PTZ convulsions in both OF1 and NMRI mice. These results suggest that besides the GABAA receptor complex other mechanisms related to calcium mobilization may be involved in the convulsant action of lindane.

c-fos and ornithine decarboxylase gene expression in brain as early markers of neurotoxicity.

An increase of proto-oncogene c-fos expression in cerebral cortex of rats treated with subconvulsant doses of the pesticide organochlorine lindane (gamma-hexachlorocyclohexane) has been detected using Northern blots. Immunohistochemical studies show that Fos protein was already increased in neuronal nuclei 3 h after treatment. The administration of the benzodiazepine diazepam prior to lindane totally blocked the activation of this proto-oncogene expression. Parallel to this increased expression of c-fos an activation of the ornithine decarboxylase (ODC) gene and enzyme was also observed. High levels of ODC mRNA and increased enzyme activity in cortex were found in rats following lindane treatment. These changes were attenuated by prior treatment of animals with diazepam. The co-induction of c-fos and ODC suggests a potential link between the ODC/polyamine system and the short-acting proto-oncogenes in stimulus-transcription coupling events.