Recalculating the staff required to run a modern assisted reproductive technology laboratory.

STUDY QUESTION: What number of staff is sufficient to perform increasingly complicated processes in today's modern ART laboratories? SUMMARY ANSWER: The adequate number of personnel required for the efficient and safe operation of modern ART laboratories needs to be calculated. WHAT IS KNOWN ALREADY: In today's modern ART laboratories, the amount of time required to perform increasingly complicated processes has more than doubled, with a downward trend in the amount of work an embryologist can do. Different workload unit values have been used to evaluate each workload task and efficiency in a particular ART laboratory, as well as to occasionally compare one laboratory with another. STUDY DESIGN, SIZE, DURATION: Seven senior embryologists working at different IVF centers, three public and four private centers, participated in this multicenter study conducted between 2019 and 2020. We prepared a survey to create a calculator for staff using the average (of three attempts) time spent in every laboratory by each embryologist of the center to perform any ART process. PARTICIPANTS/MATERIALS, SETTING, METHODS: Different laboratory processes and activities related to quality control, time spent and conventional human double witnessing were included in the survey. To calculate the number of processes that each embryologist can perform per year, an embryologist was considered to be having a full-time contract and working 7 or 8 h/day. The times included in the calculation of each task were those corresponding to the 95th percentile. For the calculations, Microsoft® Office Excel® Professional Plus 2019 was used. MAIN RESULTS AND THE ROLE OF CHANCE: The survey showed that the time needed per embryologist to perform the different processes necessary for a classic IVF cycle without time lapse (TL) was 8.11 h, and with TL, it was 10.27 h. The calculated time also considered the time spent in documentation handling, cycle preparation, database management and conventional human double witnessing verification. An ICSI without TL needed 8.55 h, and with TL, it needed 10.71 h. An ICSI-PGT without a TL cycle needed 11.75 h, and with TL, it needed 13.91 h. Furthermore, 1.81 h should be added for every vitrification support needed. The time needed to control more than 200 critical steps, including equipment control and culture parameters, was 30 min per day plus 3.9 min per device to control.The time spent in semen analysis (including documentation handling, cycle preparation and database management) or intrauterine insemination with a partner sperm was 2.7 h. For donor sperm, an additional hour was required for the management involved. The time required to perform a testicular biopsy and cryopreserve the sample was 4 h. Similarly, the time required to perform seminal cryopreservation was 3.7 h. LIMITATIONS, REASONS FOR CAUTION: The study was conducted considering a full-time contract embryologist working 7 or 8 h/day, 5 days a week, with days off according to the Spanish regulations. However, our findings can be adapted to foreign regulations using the developed online calculation platform. WIDER IMPLICATIONS OF THE FINDINGS: A new advanced staff calculator allows any IVF laboratory to estimate the minimum number of embryologists necessary without compromising the security or success of the results. Nevertheless, we recommend a minimum of two qualified embryologists in every laboratory, regardless of the workload. STUDY FUNDING/COMPETING INTEREST(S): This work was funded by the Asociación para el Estudio de la Biología de la Reproducción (ASEBIR). None of the authors has any conflict of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Potentially inappropriate medications in older adults living with HIV.

OBJECTIVES: We assessed the prevalence of potentially inappropriate medication (PIM) among older (≥ 65 years) people living with HIV (O-PLWH) in the region of Madrid. METHODS: We analysed the dispensation registry of community and hospital pharmacies from the Madrid Regional Health Service (SERMAS) for the period between 1 January and 30 June 2017, looking specifically at PIMs according to the 2019 Beers criteria. Co-medications were classified according to the Anatomical Therapeutic Chemical (ATC) classification system. RESULTS: A total of 6 636 451 individuals received medications. Of these individuals, 22 945 received antiretrovirals (ARVs), and of these 1292 were O-PLWH. Overall, 1135 (87.8%) O-PLWH were taking at least one co-medication, and polypharmacy (at least five co-medications) was observed in 852 individuals (65.9%). A PIM was identified in 482 (37.3%) O-PLWH. Factors independently associated with PIM were polypharmacy [adjusted odds ratio (aOR) 7.08; 95% confidence interval (CI) 5.16-9.72] and female sex (aOR 1.75; 95% CI 1.30-2.35). The distribution of PIMs according to ATC drug class were nervous system drugs (n = 369; 28.6%), musculoskeletal system drugs (n = 140; 10.8%), gastrointestinal and metabolism drugs (n = 72; 5.6%), cardiovascular drugs (n = 61; 4.7%), respiratory system drugs (n = 13; 1.0%), antineoplastic and immunomodulating drugs (n = 10; 0.8%), and systemic anti-infectives (n = 2; 0.2%). Five drugs accounted for 84.8% of the 482O PLWH with PIMs: lorazepam (38.2%), ibuprofen (18.0%), diazepam (10.2%), metoclopramide (9.9%), and zolpidem (8.5%). CONCLUSIONS: Prescription of PIMs is highly prevalent in O-PLWH. Consistent with data in uninfected elderly people, the most frequently observed PIMs were benzodiazepines and nonsteroidal anti-inflammatory drugs . Targeted interventions are warranted to reduce inappropriate prescribing and polypharmacy in this vulnerable population.

Activated carbon from flash pyrolysis of eucalyptus residue.

Forestry waste (eucalyptus sp) was converted into activated carbon by initial flash pyrolysis followed carbonization and CO2 activation. These residues were obtained from a pilot plant in Spain that produces biofuel, the biochar represented 10-15% in weight. It was observed that the highest activation was achieved at a temperature of 800 °C, the specific surface increased with time but, on the contrary, high loss of matter was observed. At 600 °C, although there was an important increase of the specific surface and the volume of micropores, at this temperature it was observed that the activation time was not an influential parameter. Finally, at 400 °C it was observed that the activation process was not very significant. Assessing the average pore diameter it was found that the lowest value corresponded to the activation temperature of 600 °C, which indicated the development of microporosity. When the activation temperature increases up to 800 °C the pore diameter increased developing mesoporosity.

Polymorphisms in Host Immunity-Modulating Genes and Risk of Invasive Aspergillosis: Results from the AspBIOmics Consortium.

Recent studies suggest that immune-modulating single-nucleotide polymorphisms (SNPs) influence the risk of developing cancer-related infections. Here, we evaluated whether 36 SNPs within 14 immune-related genes are associated with the risk of invasive aspergillosis (IA) and whether genotyping of these variants might improve disease risk prediction. We conducted a case-control association study of 781 immunocompromised patients, 149 of whom were diagnosed with IA. Association analysis showed that the IL4Rrs2107356 and IL8rs2227307 SNPs (using dbSNP numbering) were associated with an increased risk of IA (IL4Rrs2107356 odds ratio [OR], 1.92; 95% confidence interval [CI], 1.20 to 3.09; IL8rs2227307 OR, 1.73; 95% CI, 1.06 to 2.81), whereas the IL12Brs3212227 and IFNγrs2069705 variants were significantly associated with a decreased risk of developing the infection (IL12Brs3212227 OR, 0.60; 95% CI, 0.38 to 0.96; IFNγrs2069705 OR, 0.63; 95% CI, 0.41 to 0.97). An allogeneic hematopoietic stem cell transplantation (allo-HSCT)-stratified analysis revealed that the effect observed for the IL4Rrs2107356 and IFNγrs2069705 SNPs was stronger in allo-HSCT (IL4Rrs2107356 OR, 5.63; 95% CI, 1.20 to 3.09; IFNγrs2069705 OR, 0.24; 95% CI, 0.10 to 0.59) than in non-HSCT patients, suggesting that the presence of these SNPs renders patients more vulnerable to infection, especially under severe and prolonged immunosuppressive conditions. Importantly, in vitro studies revealed that carriers of the IFNγrs2069705C allele showed a significantly increased macrophage-mediated neutralization of fungal conidia (P = 0.0003) and, under stimulation conditions, produced higher levels of gamma interferon (IFNγ) mRNA (P = 0.049) and IFNγ and tumor necrosis factor alpha (TNF-α) cytokines (P value for 96 h of treatment with lipopolysaccharide [PLPS-96 h], 0.057; P value for 96 h of treatment with phytohemagglutinin [PPHA-96 h], 0.036; PLPS+PHA-96 h = 0.030; PPHA-72 h = 0.045; PLPS+PHA-72 h = 0.018; PLPS-96 h = 0.058; PLPS+PHA-96 h = 0.0058). Finally, we also observed that the addition of SNPs significantly associated with IA to a model including clinical variables led to a substantial improvement in the discriminatory ability to predict disease (area under the concentration-time curve [AUC] of 0.659 versus AUC of 0.564; P-2 log likehood ratio test = 5.2 · 10(-4) and P50.000 permutation test = 9.34 · 10(-5)). These findings suggest that the IFNγrs2069705 SNP influences the risk of IA and that predictive models built with IFNγ, IL8, IL12p70, and VEGFA variants can used to predict disease risk and to implement risk-adapted prophylaxis or diagnostic strategies.

La investigación biomédica hospitalaria a través de la satisfacción de los profesionales de un Instituto de Investigación Sanitaria / Hospital biomedical research through the satisfaction of a Health Research Institute professionals

Objetivo. Los Institutos de Investigación Sanitaria son una apuesta estratégica de gran alcance para impulsar la investigación biomédica en los hospitales. Evaluar la satisfacción de sus usuarios es un requisito de calidad imprescindible. El objetivo de este trabajo es evaluar la satisfacción de los profesionales de un Instituto de Investigación Sanitaria, centro de investigación biomédica hospitalaria por excelencia. Métodos. Estudio observacional a través de un cuestionario de satisfacción a los investigadores de un Instituto de Investigación Sanitaria. Las dimensiones exploradas derivaron de los servicios ofrecidos a los investigadores y que se articulan en torno a los 4 ejes del Plan Estratégico quinquenal. Se realizó estudio descriptivo y analítico según las variables de ajuste. Se calculó la consistencia interna del cuestionario. Resultados. La encuesta fue completada por 108 investigadores (15% de respuesta). El aspecto estratégico mejor valorado fue la estructuración en Áreas/Grupos de Investigación y la política de comunicación. La valoración global fue de 7,25 sobre 10. Las sugerencias de mejora giran en torno a la necesidad de ayudas para la contratación y las infraestructuras de investigación. El cuestionario ha resultado tener una alta consistencia interna (alfa de Cronbach de 0,9). Conclusiones. Hasta el momento las políticas de investigación en el entorno sanitario y biomédico no han sido suficientemente valoradas por los profesionales en nuestro ámbito. Sistematizar la evaluación de la satisfacción y las expectativas de los principales grupos de interés es un instrumento esencial de análisis, participación en la mejora continua y de avanzar hacia la excelencia de la investigación sanitaria (AU)

Objective. A Health Research Institute is a powerful strategic commitment to promote biomedical research in hospitals. To assess user satisfaction is an essential quality requirement. The aim of this study is to evaluate the professional satisfaction in a Health Research Institute, a hospital biomedical research centre par excellence. Methods. Observational study was conducted using a satisfaction questionnaire on Health Research Institute researchers. The explored dimensions were derived from the services offered by the Institute to researchers, and are structured around 4 axes of a five-year Strategic Plan. A descriptive and analytical study was performed depending on adjustment variables. Internal consistency was also calculated. Results. The questionnaire was completed by 108 researchers (15% response). The most valued strategic aspect was the structuring Areas and Research Groups and political communication and dissemination. The overall rating was 7.25 out of 10. Suggestions for improvement refer to the need for help in recruitment, and research infrastructures. High internal consistency was found in the questionnaire (Cronbach alpha of 0.9). Conclusions. So far research policies in health and biomedical environment have not been sufficiently evaluated by professionals in our field. Systematic evaluations of satisfaction and expectations of key stakeholders is an essential tool for analysis, participation in continuous improvement and advancing excellence in health research (AU)

[Hospital biomedical research through the satisfaction of a Health Research Institute professionals]. / La investigación biomédica hospitalaria a través de la satisfacción de los profesionales de un Instituto de Investigación Sanitaria.

OBJECTIVE: A Health Research Institute is a powerful strategic commitment to promote biomedical research in hospitals. To assess user satisfaction is an essential quality requirement. The aim of this study is to evaluate the professional satisfaction in a Health Research Institute, a hospital biomedical research centre par excellence. METHODS: Observational study was conducted using a satisfaction questionnaire on Health Research Institute researchers. The explored dimensions were derived from the services offered by the Institute to researchers, and are structured around 4 axes of a five-year Strategic Plan. A descriptive and analytical study was performed depending on adjustment variables. Internal consistency was also calculated. RESULTS: The questionnaire was completed by 108 researchers (15% response). The most valued strategic aspect was the structuring Areas and Research Groups and political communication and dissemination. The overall rating was 7.25 out of 10. Suggestions for improvement refer to the need for help in recruitment, and research infrastructures. High internal consistency was found in the questionnaire (Cronbach alpha of 0.9). CONCLUSIONS: So far research policies in health and biomedical environment have not been sufficiently evaluated by professionals in our field. Systematic evaluations of satisfaction and expectations of key stakeholders is an essential tool for analysis, participation in continuous improvement and advancing excellence in health research.

Perioperative values of glutathione peroxidase activity and malondialdehyde levels in enolic cirrhotic recipients of a liver transplant.

Reactive oxygen species play a central role in ischemia-reperfusion injury after organ transplantation. They are degraded by endogenous radical scavengers such as antioxidant enzymes. The purpose of this study was to evaluate the temporal variation in glutathione peroxidase (GPX) activity and malondialdehyde (MDA) levels among alcoholic cirrhotic recipients of liver transplantations. The study included 30 recipients: 26 males and 4 females in the provided blood samples before and after transplantation. The results showed significant enhancement of MDA levels at 1 and 6 hours after transplantation: 4.458 ± 2.273 µmol/L and 4.4628 ± 2.405 µmol/L respectively (P < .001). In contrast, GPX activity showed a maximum at 3 days there after 3.541 ± 2,315 nmol/mg protein. In conclusion, although MDA levels show an enormous increase at 1 hour after transplantation suggesting lipid peroxidation, they were compensated by GPX activity thereafter, indicating control of the oxidative stress generated by liver transplantation.

Echocardiographic study in cirrhotic patient candidates for liver transplantation.

We studied 81 cirrhotic patients who were candidates for liver transplantation to evaluate frequently detected cardiac alterations by echocardiographic study. Patients were distributed into three groups: group 1 comprised alcoholic cirrhotic patients (n = 40); group 2, viral cirrhotic patients (hepatitis C or B virus) (n = 35); and group 3, patients with primary biliary cirrhosis (n = 6). Cardiac chambers and diastolic functions were estimated by two-dimensional transthoracic echocardiography in M mode and Doppler. The most frequently detected cardiac structural alterations were left atrial diameter enlargement in 100% of the women and 40% of the men in group 1; 87.5% of the women and 15.4% of the men in group 2; and 33.3% of the women in group 3. Interventricular wall thickness enlargement in 50% of the women and 27.8% of the men in group 1, 25% of the women and 30.8% of the men in group 2, and 16.4% of the women in group 3. The prevalence of diastolic dysfunction was 45% in group 1, 32.3% in group 2, and 16.4% in group 3 (P > .05). There were no significant differences between the groups in cardiac chamber dimensions, left ventricular wall thickness, or prevalence of diastolic dysfunction.

Gene network profiling before and after transplantation in alcoholic cirrhosis liver transplant recipients.

The main objective of this study was to define a gene network profile network in liver transplant recipients with alcoholic cirrhosis before and after liver transplantation. Genes were selected from data obtained in a previous study of liver transplant recipients with alcoholic cirrhosis. Selected up-regulated genes were further validated by quantitative real-time polymerase chain reaction in different groups of liver transplant recipients with alcoholic cirrhosis (n=5). Selected genes up-regulated before transplantation were: TNFRSF9 (tumor necrosis factor [TNF] receptor superfamily, member 9); IL2RB (interleukin-2 receptor beta); BCL2L2 (BCL2-like 2); NOX5 (NADPH) oxidase, EF-hand calcium binding domain 5); PEX5 (peroxisomal biogenesis factor 5); PPARG (peroxisome proliferator-activated receptor gamma); NIBP (IKK2 binding protein); NKIRAS2 (NFKappaBeta inhibitor interacting Ras-like 2); IL4 (interleukin-4); IL-4R (interleukin 4 receptor); ADH1A (alcohol dehydrogenase 1A, class 1); ALDH1L1 (aldehyde dehydrogenase 1 family, member L1); MPO (myeloperoxidase); NPPA (natriuretic peptide precursor A); BCL2A1 (BCL2-related protein A1); GADD45A (growth arrest and DNA-damage-inducible alpha); TEGT (Bax inhibitor 1); PIK3CA (phosphoinositide-3-kinase, catalytic, alpha polypeptide); IFNGR2 (interferon gamma receptor 2); JAK2 (Janus Kinase 2); FAS (Fas, TNF receptor superfamily, member 6); TANK (TRAF family member-associated NFKB activator); TTRAP (TRAF and TNF receptor-associated protein); and ANXA5 (annexin A5).

Hepatopulmonary syndrome among cirrhotic candidates for liver transplantation.

This study assess of hepatopulmonary syndrome (HPS) prevalence and the influence of etiology among cirrhotic patients due to an alcoholic or viral etiology. We examined the records of patients were distributed as Group 1, alcoholic (n = 40) and Group 2, hepatic cirrhosis of viral etiology (n = 35). Hepatic cirrhosis status was estimated by CHILD and MELD scores. Presence of clinical ascites spell out was noted as well as size and diastolic functions of the cardiac chambers using two-dimensional transthoracic echocardiography in M mode and by Doppler. HPS was studied with agitated saline serum and intravenous contrast administration. HPS was considered to be present when serum or contrast passed to the left chamber before the 5th cardiac cycle. There was no significant differences among related to sex, age, cirrhosis status or ascites. HPS frequency was 35% in Group 1 versus 64.7% among Group 2-Patients (P = .01). Taking into account the results, we concluded that HPS frequency was related to cirrhotic etiology. Upon multivariate analysis a patients with cirrhosis from viral etiology showed significantly increased HPS frequency compared with those displaying cirrhosis of an alcoholic etiology.

Perioperative levels of glutathione reductase in liver transplant recipients with hepatitis C virus cirrhosis.

Surgical intervention causes oxidative stress leading to an adaptive responses by the body. To evaluate changes in the defense capacity of antioxidant enzymes, we determined the activity of glutathione reductase (GR) levels among liver transplant recipients with due to hepatitis C virus cirrhosis. The study was performed in 22 patients (16 males and 6 females) of average ages 52.63 ± 5.49 years for males and 59.67 ± 5.65 years for females. Blood samples for glutathione reductase activity were drawn on admission before as well as at 1, 6, and 12 h and 1, 2, 3, 5 and 7 days after the liver transplantation. Perioperative glutathione reductase levels were significant (P = .014) over the period using Bonferroni tests. GR activity reached a maximum (15.6112 ± 6.56035 nmol/mg protein) at 3 days after liver transplantation (T3d) (P = .001). The increased GR activity values detected perioperatively indicated scavenging of reactive oxygen species generated after liver transplantation of hepatitis C virus cirrhosis patients.

Determination of the risk of self-ignition of coals and biomass materials.

The safe storage, processing and handling of coals and biomass resources requires their tendency to self-ignite be understood; fires caused by self-ignition have occurred on many occasions in ports and at industrial plants. This work provides information on the tendency of several types of coal and four types of biomass to self-ignite. Data were obtained using the isothermal oven procedure and analyzed using the Frank-Kamenetskii method and a scaling procedure, both contemplated in standard EN15188. The results obtained throw light on the optimum volumes and storage times of the studied materials. The results also validate the methodology followed for determining the risk of self-ignition.

Preoperative natriuretic peptide-B values and ascites in male liver transplant recipients.

Plasma preoperative values of natriuretic B peptide (pro-BNP) were correlated with ascites in men experiencing hepatic cirrhosis due to different etiologies on the active waiting list for liver transplantation. The study was performed in 54 male recipients of a liver transplant. Written informed consent was obtained from the patients or their relatives, and the study protocol was approved by our local Clinical Research (Ethics) Committee. Male patients were classified into two groups: group 1 included patients with alcoholic hepatic cirrhosis (n = 30) distributed as 19 men with no ascites, four with nonrefractory ascites, and seven with refractory ascites; group 2 included cases of viral hepatitis cirrhosis (n = 24) distributed as 13 men with no ascites, nine with non-refractory ascites, and two with refractory ascites. A group of six healthy male volunteers was used to establish normal (basal) values of pro-BNP and left auricular diameter (LAD). Pro-BNP values were determined in plasma samples by an electrochemiluminiscence immunoassay. Pro-BNP plasma levels in patients with alcoholic cirrhosis were threefold greater among patients with no ascites or no refractory ascites compared with healthy men, whereas pro-BNP values were fivefold enhanced among alcoholic patients with refractory ascites. The viral hepatitis cirrhosis group showed pro-BNP plasma values 1.5-fold enhanced in men with no ascites, whereas pro-BNP reached fivefold with either nonrefractory or refractory ascites. The enhanced pro-BNP plasma levels indicated advanced hepatic degradation, seemingly related to the presence of refractory ascites associated with cirrhosis.

Intraoperative pH values after N-acetylcysteine administration during liver transplantation.

We investigated whether intraoperative administration of N-acetylcysteine (NAC) to liver transplant recipients affected pH values. This prospective, randomized, double-blind clinical trial included liver transplant recipients who were randomly assigned to NAC-treated (n=25) or placebo (n=25) groups. The NAC-treated group received 100 mg/kg dissolved in 5% dextrose over 15 minutes during the anhepatic phase, followed by a continuous infusion of 50 mg/kg in 5% dextrose during the next 24 hours. The placebo group received equal amounts of 5% dextrose solution during the same times. Peripheral blood samples were drawn in Ca2+-80 IU-containing syringes after induction of anesthesia (I-1), at 15 minutes into the anhepatic phase (I-2) prior to the administration of NAC or placebo, at 5 minutes before reperfusion (I-3), at 5 minutes after reperfusion (I-4), at 20 minutes after reperfusion (I-5), at 60 minutes after reperfusion (I-6), and at 1 hour after completion of the procedure (I-7). pH levels, which were determined using a radiometer ABL77 (Copenhagen, Denmark), were significantly lower among the NAC than the placebo group at I-4 (P=.027) and I-5 (P=.031). An early decrease in pH values was detected in the NAC-treated group at 5 minutes before reperfusion (I-3; P=.051). We concluded that intraoperative NAC administration during the anhepatic phase of liver transplantation significantly decreased recipient pH values at 5 and 20 minutes after reperfusion, a decrease that was detected at 5 minutes before reperfusion (I-3). The decrease seemed to be associated with NAC metabolism.

TNFR1 mRNA expression level and TNFR1 gene polymorphisms are predictive markers for susceptibility to develop invasive pulmonary aspergillosis.

Tumour necrosis factor (TNF) is primarily secreted by monocytes/macrophages and activated T lymphocytes in response to fungal infections. TNF acts through TNF receptor 1 (TNFR1) triggering a pro-inflammatory response, and therefore plays a pivotal role in immune regulation and host immune responses. We hypothesized that single nucleotide polymorphisms (SNPs) in TNFR1 gene may influence the innate immune response against Aspergillus. Three SNPs were genotyped in 275 individuals (144 immunocompromised haematological patients with high-risk of developing IPA and 131 healthy controls): TNFR1(-383(A/C)) (rs2234649) and TNFR1(-609(G/T)) (rs4149570) in the 5 prime UTR region, and TNFR1(+36(A/G)) SNP (rs767455) in the first exon of the gene. Of the 144 haematological patients, 77 patients developed Invasive Pulmonary Aspergillosis (IPA) infection and the remaining 67 patients were not infected. TNFR1(+36(A/G)) and TNFR1(-609(G/T)) were associated with IPA susceptibility (p=0.033 and p=0.018, respectively). A role of TNFR1 genetic variants in the susceptibility of patients to develop IPA was also supported by the significantly lower TNFR1 mRNA expression level in IPA than in IPA-resistant patients and the strong correlation between the TNFR1(-609) genetic variant and the expression levels of TNFR1. There was also a tendency for a higher frequency of galactomannan (GM) positivity in patients with TNFR1(-609G/G) genotype than in patients with TNFR1(-609G/T) (p=0.0909) or TNFR1(-609T/T) (p=0.0913) genotype. Predictive sequence analysis of the effects of TNFR1(-609) promoter polymorphism revealed that this SNP might play a critical role in modifying the affinity of ICSBP/IRF-8, a transcription factor that is involved in the TNFR1-mediated activation of NFkappaB signalling pathway. Taken together, these data suggest that TNFR1 polymorphisms influence the risk of IPA disease and might be useful for risk stratification strategies. These findings need to be confirmed in validation studies with larger samples of haematological patients.

A study of F-waves in patients with unilateral lumbosacral radiculopathy.

BACKGROUND AND PURPOSE: The F wave, a late response of low amplitude, is widely used in the study of peripheral nerve lesions, and its persistence and latencies are the main parameters that are usually considered. The analysis of repeater F-waves, which are commonly observed in association with focal or generalized motor neuropathy, is not always performed as a standard electrodiagnostic protocol. METHODS: We recorded and quantified the F waves from 13 healthy subjects and 22 patients with unilateral lumbosacral radiculopathy (ULSR) affecting the L5 or S1 roots. RESULTS: We found differences between the injured and normal sides of patients with ULSR in several F-wave parameters. Taking into consideration the normalized and pooled values of tibial and peroneal nerves in the injured side of patients with ULSR, the minimum and mean latencies were higher (1.05 and 1.04 with respect to 1.00; P < 0.01), the relative amplitude of the F waves was higher (1.95 with respect to 1.00; P < 0.001), and the percentage of repeater F-waves was also higher (4.19 with respect to 1.00; P < 0.001). This latter parameter was the most sensitive to detect lateral differences as indicated by the percentage of change and its high z score. CONCLUSIONS: Our results show that the use of F-waves may improve the electrodiagnosis of the ULSR if the number of repeater waves is evaluated given the clear and consistent increase of this variable in patients with lumbosacral root injury.

Gene expression profiling in liver transplant recipients with alcoholic cirrhosis.

The main objective of this study was to identify differences in gene expression profile using microarray technology in liver transplant recipients with alcoholic cirrhosis before and after liver transplantation. The study was performed in liver transplant recipients with alcoholic cirrhosis (n = 10) and in healthy volunteers (n = 10), as a reference group. Peripheral blood samples were obtained before (T0) and 7 days after liver transplantation (T7d) using tubes with an RNA stabilizer. RNA was purified and quality tested. From each participant in the study, microarrays were done in duplicate using 10 mug of cRNA. After reverse transcription, complementary RNAs were labeled with Cy5 Streptavidine and used for hybridization of 20,000 human genes CodeLink bioarrays (Applied Microarrays, United States) overnight at 37 degrees C. Arrays were read with a laser scanner and quantified and normalized with CodeLink Software 4.2. Liver transplant recipients showed a gene expression profile before transplantation (T0) of 4310 overexpressed genes compared with healthy volunteers, with 407 of these genes increased more than 2-fold (P < .05). After transplantation (T7d), the same group of patients showed a profile of 1011 overexpressed genes compared with T0, with 109 of these genes increased more than 2-fold (P < .05). We determined gene expression profiles in peripheral blood samples obtained before and after liver transplantation, giving a report of array gene expression profiles of peripheral blood samples from each of these patients. One implication of these results is that gene profiling of peripheral blood samples using microarray technology could be used to dynamically monitor the impact and adequacy of immunosuppression in individual patients.

Study of gene expression profile in liver transplant recipients with hepatitis C virus.

The main objective of this study was to identify differences in gene expression profiles by liver transplant recipients with hepatitis C virus (HCV) using microarray technology before versus after liver transplantation. The study was performed in liver transplant recipients with HCV (n = 6) versus a group of healthy volunteers (n = 6). Peripheral blood samples were obtained before (T0) and 7 days after liver transplantation (T7d) using tubes with an RNA stabilizer. The quality of purified RNA was tested (28S/18S ratio >1.5) in a bioanalyzer. Each participant in the study underwent microarrays in duplicate using 10 mug of complementary RNA. After reverse transcription, cRNAs were labeled with Cy5 Streptavidine. Hybridization of 20000 human genes CodeLink bioarrays (Applied Microarrays, United States) was performed overnight at 37 degrees C. Arrays read with a laser scanner were normalized with CodeLink Software 4.2. At T0, liver transplant recipients showed 116 over-expressed genes when compared with healthy volunteers, who had 33 genes increased >2-fold (P < .05). At T7d after transplantation, the same group of patients showed 613 over-expressed genes compared with T0, of which 97 genes were increased >2-fold (P < .05). We determined gene expression profiles in peripheral blood samples obtained before and after liver transplantation, reporting the array of gene expression profiles in peripheral blood samples from each of these patients classes. One implication of these results is that gene profiling of peripheral blood samples could be used to dynamically monitor the impact and adequacy of immunosuppression in individual patients using microarray technology.

Effect of N-acetylcysteine administration on intraoperative plasma levels of interleukin-4 and interleukin-10 in liver transplant recipients.

We investigated whether intraoperative administration of N-acetylcysteine (NAC) in liver transplant recipients ameliorated their inflammatory responses by increasing intraoperative plasma levels of interleukin (IL)-4 and IL-10. This prospective, randomized, double-blind clinical trial included liver transplant recipients randomly assigned to the NAC-treated (n = 25) or the placebo (n = 25) group. The NAC-treated group received 100 mg/kg dissolved in 5% dextrose over 15 minutes during the anhepatic phase, followed by a continuous infusion of 50 mg/kg in 5% dextrose over the next 24 hours, whereas the placebo group received equal amounts of 5% dextrose solution during the same time. Peripheral blood samples were drawn in EDTA-containing tubes after induction of anesthesia (I-1); at 15 minutes into the anhepatic phase (I-2) prior to the administration of NAC or placebo; at 5 minutes before reperfusion (I-3); at 10 minutes after reperfusion (I-4); at 20 minutes after reperfusion (I-5); at 60 minutes after reperfusion (I-6); and at 1 hour after completion of the liver transplantation (I-7). Cytokine levels were determined using a technique which combined enzyme-linked immunosorbent assay (ELISA) and flow cytometry. Plasma IL-4 levels were significantly higher among the NAC-treated group than the placebo group at I-3 (P = .046) and I-4 (P = .041). Plasma IL-10 levels showed significant enhancement in the NAC-treated group at 5 minutes before reperfusion (I-3; P = .007). We concluded that intraoperative NAC administration during the anhepatic phase of liver transplantation significantly increased recipient IL-4 plasma levels before and after reperfusion, and IL-10 plasma values before reperfusion (I-3). These enhancements seemed to be associated with a protective effect against reperfusion injury.