Profiling of drug-metabolizing enzymes/transporters in CD33+ acute myeloid leukemia patients treated with Gemtuzumab-Ozogamicin and Fludarabine, Cytarabine and Idarubicin.

Genetic heterogeneity in drug-metabolizing enzyme/transporter (DMET) genes affects specific drug-related cancer phenotypes. To investigate the relationships between genetic variation and response to treatment in acute myeloid leukemia (AML), we genotyped 1931 variants on DMET genes in 94 CD33-positive AML patients enrolled in a phase III multicenter clinical trial combining Gemtuzumab-Ozogamicin (GO) with Fludarabine-Cytarabine-Idarubicin (FLAI) regimen, with the DMET Plus platform. Two ADH1A variants showed statistically significant differences (odds ratio (OR)=5.68, P=0.0006; OR=5.35, P=0.0009) in allele frequencies between patients in complete/partial remission and patients without response, two substitutions on CYP2E1 (OR=0.13, P=0.001; OR=0.09, P=0.003) and one on SLCO1B1 (OR=4.68, P=0.002) were found to differently influence liver toxicity, and two nucleotide changes on SULTB1 and SLC22A12 genes correlated with response to GO (OR=0.24, P=0.0009; OR=2.75, P=0.0029). Genetic variants were thus found for the first time to be potentially associated with differential response and toxicity in AML patients treated with a combination of GO-FLAI regimen.

Analysis of nosocomial acquired Clostridium difficile infection in an Italian research and teaching hospital.

BACKGROUND: Clostridium difficile (Cd) infection is a nosocomial plague which is correlated with several clinical and medical factors such as antibiotics intake. It is known that prevention is possible through infection control measures both clinical and epidemiological. METHODS: We examined the data from a study about Cd infection in four internal medicine wards in a teaching and research hospital in the north part of Italy in a two years period. The wards are only slightly different in size, plan, structures, nursing staff and patient's characteristics but have a different room' organization, lay out and different level of continuous education programs for nursing personnel. RESULTS: We reported a high incidence of the infection and a non-significant difference between wards also looking to the different possibility-capacity of taking preventive measures and the different level of nursing staff continuous educational performance. CONCLUSION: The analysis of the data we obtained was the basis to write a protocol and to start a training course for the medical and nursing personnel of the four wards on the managing of patients infected with Cd infection. On March 2011 we started a one year longitudinal study about the Cd infections in the same wards with the purpose of evaluating the adherence to the protocol, monitoring the incidence of infection and studying the risk factors of the infected patients related to the proper use of the protocol on Cd.

Magnetoencephalography reveals differences in brain activations for fast and slow responses to simple multiplications.

Despite decades of studies, it is still an open question on how and where simple multiplications are solved by the brain. This fragmented picture is mostly related to the different tasks employed. While in neuropsychological studies patients are asked to perform and report simple oral calculations, neuroimaging and neurophysiological studies often use verification tasks, in which the result is shown, and the participant must verify the correctness. This MEG study aims to unify the sources of evidence, investigating how brain activation unfolds in time using a single-digit multiplication production task. We compared the participants' brain activity-focusing on the parietal lobes-based on response efficiency, dividing their responses in fast and slow. Results showed higher activation for fast, as compared to slow, responses in the left angular gyrus starting after the first operand, and in the right supramarginal gyrus only after the second operand. A whole-brain analysis showed that fast responses had higher activation in the right dorsolateral prefrontal cortex. We show a timing difference of both hemispheres during simple multiplications. Results suggest that while the left parietal lobe may allow an initial retrieval of several possible solutions, the right one may be engaged later, helping to identify the solution based on magnitude checking.

Clinical outcomes of Clostridium difficile infection according to strain type. A prospective study in medical wards.

OBJECTIVES: To describe clinical characteristics and outcome of Clostridium difficile infection (CDI) patients in Internal Medicine, to identify ribotypes (RTs); to evaluate the association between RT and patient clinical characteristics and report outcome. METHODS: One year prospective cohort study. Clinical data, Barthel Index (BI) and outcomes were collected for all inpatients suffering from CDI (n = 148) in hospital wards in Northern Italy. 84 fecal samples were analysed for molecular typing. RESULTS: 12 RTs were identified, predominantly RT018 (42.9%, n = 36/84) and RT356/607 (40.5%, n = 34/84). Patients with dementia were more frequent among those infected by RT018 [55.6% (n = 20/36) vs. 32.4% (n = 11/34), p = 0.05]. The median BI score of patients with RT018 was lower than BI score of patients with RT356/607 [10 (IQR 0-32) vs. 15 (IQR 5-50), p = 0.06]. RT018 infection was associated to higher levels of C-reactive protein [7.2 mg/dl (IQR 4.1-14.7) vs. 4.0 mg/dl (IQR 2.2-6.8), p = 0.01] and white blood cells ≥15,000/dl [33.3% (n = 12/36) vs. 14.7% (n = 5/34) of patients, p = 0.07]. Higher mortality was noted among RT018 infected patients. We found a continuous mortality increase according to the ATLAS score. CONCLUSIONS: Our results confirm that RT018 and RT356/607 are the two major RTs causing CDI in older patients with a high degree of disability in Northern Italy and RT018 is associated with more serious outcomes.

Clostridium difficile infection epidemiology and management: Comparison of results of a prospective study with a retrospective one in a reference teaching and research hospital in Northern Italy.

BACKGROUND: Clostridium difficile-associated disease (CDAD) is the most common infectious antibiotic-associated diarrhea and is a growing health care problem. Prevention of Clostridium difficile infection focuses on clinical and epidemiologic infection control measures. METHODS: Between 2008 and 2009, we conducted a retrospective study that showed an incidence of CDAD among the highest reported in the literature. Subsequently, we developed a preventive protocol that was adopted in our hospital in 2010. We then conducted a prospective study to investigate prevalence, incidence, and mortality of CDAD and to compare the results with those of the retrospective study, evaluating adherence to preventive measures and their efficacy. RESULTS: In both studies, prevalence and incidence significantly increased in older patients. Crude prevalence was similar in the 2 studies. The incidence rate increased by 36%, with a significant increase only in the C and D wards. In-hospital mortality rose in both prevalent and incident cases. Regarding adhesion to hospital protocol, 77% of prevalent cases were treated with the required procedure. The highest percentage of isolated patients was achieved in C and D wards. In these wards we detected lower training hours per nurse. However, in 2013, we observed a significant decrease in incidence of CDAD and found a hospital prevalence of 0.33%. CONCLUSIONS: Health care personnel education could be more important than the possibility of isolating infected patients in single rooms.

Estimating bulk density in vertically exposed stoney alluvium using a modified excavation method.

Despite many decades of education and refining land-use practices, accelerated stream bank erosion is still prevalent in the United States. Eroding stream banks produce a sediment load to the riverine system and can cause reduced water quality as a result of increased suspended sediment. As total maximum daily loads (TMDLs) for water bodies impaired by turbidity or suspended sediments become more numerous, a simple, in situ field technique will be needed to estimate the bulk density of readily erodible stream bank material so that reasonably accurate sediment loading rates can be estimated. In this study, the excavation/polyurethane-foam technique for estimating total bulk density was applied to vertically exposed alluvium with high coarse-fragment content. Though not previously attempted in vertically exposed alluvium with high coarse-fragment content, the excavation/polyurethane-foam technique appears to provide a reasonably accurate estimate of the total and soil (<2-mm size fraction) bulk density from vertically exposed, alluvial deposits with high coarse-fragment content (i.e., >70%) along eroding stream banks. Obtaining bulk density estimates using this method would facilitate calculation of sediment loading rates to riverine systems with actual field data.

Microarray gene expression analysis of porcine skeletal muscle sampled at several post mortem time points.

A pilot study using Affymetrix Gene Chip(®) Porcine Genome Arrays was set up to evaluate the impact of time lags from death on gene expression profiling of porcine skeletal muscle at four post mortem times (up to 24h) during the routine processing of fresh thighs. All post chip parameters and data analyses (Average background, Scale Factors, Percent Presence, 3'/5' ratios of ß-actin and glyceraldehyde-3-phosphase dehydrogenase control genes, RNA degradation diagnostics, principal component analysis, hierarchical clustering, mixed regression models with time effects) did not show any effect of post mortem time. Therefore, microarray data obtained from muscle specimens collected in a processing plan over a quite long period have the potential to identify treatments or pre mortem conditions without any potential bias derived from subtle RNA degradation. These results open new perspectives to develop and analyse gene expression biomarkers for pig production and product authentication.

Filariasis focus due to Mansonella ozzardi and Mansonella perstans in the Amazon Federal Territory of Venezuela.

The presence of filariasis was investigated through Knott's method in 40 Curripaco Indians from communities located on the Casiquiare and Guainia Rivers in the Amazon Federal Territory of Venezuela. The results gave an 82.5% prevalence for infections with Mansonella perstans and 30% for infections with M. ozzardi (mixed infections with M. perstans). In almost all the patients the parasite load was greater for M. perstans and varied between 1 and 327 microfilariae per ml. This paper presents for the first time the presence of a transmission focus of M. perstans in the Amazon Federal Territory and confirms the finding of M. ozzardi in the region.

Interaction with extracellular matrix proteins influences Lsh/Ity/Bcg (candidate Nramp) gene regulation of macrophage priming/activation for tumour necrosis factor-alpha and nitrite release.

The murine resistance gene Lsh/Ity/Bcg regulates activation of macrophages for tumour necrosis factor-alpha (TNF-alpha)-dependent production of nitric oxide mediating antimicrobial activity against Leishmania, Salmonella and Mycobacterium. As Lsh is differentially expressed in macrophages from different tissue sites, experiments were performed to determine whether interaction with extracellular matrix (ECM) proteins would influence the macrophage TNF-alpha response. Plating of bone marrow-derived macrophages onto purified fibrinogen or fibronectin-rich L929 cell-derived matrices, but not onto mannan, was itself sufficient to stimulate TNF-alpha release, with significantly higher levels released from congenic B10.L-Lshr compared to C57BL/10ScSn (Lshs) macrophages. Only macrophages plated onto fibrinogen also released measurable levels of nitrites, again higher in Lshr compared to Lshs macrophages. Addition of interferon-gamma (IFN-gamma), but not bacterial lipopolysaccharide or mycobacterial lipoarabinomannan, as a second signal enhanced the TNF-alpha and nitrite responses of macrophages plated onto fibrinogen, particularly in the Lshr macrophages. Interaction with fibrinogen and fibronectin also primed macrophages for an enhanced TNF-alpha response to leishmanial parasites, but this was only translated into enhanced nitrite responses in the presence of IFN-gamma. In these experiments, Lshr macrophages remained superior in their TNF-alpha responses throughout, but to a degree which reflected the magnitude of the difference observed on ECM alone. Hence, the specificity for the enhanced TNF-alpha responses of Lshr macrophages lay in their interaction with fibrinogen and fibronectin ECM, while a differential nitrite response was only observed with fibrinogen and/or IFN-gamma. The results are discussed in relation to the possible function of the recently cloned candidate gene Nramp, which has structural identity to eukaryote transporters and an N-terminal cytoplasmic proline/serine-rich putative SH3 binding domain.

Increases in booster seat use among children of low income families and variation with age.

OBJECTIVES: To increase booster seat use among low income parents. DESIGN/METHODS: A pre-test/post-test design conducted in nine daycare centers with post-test observations four to eight weeks after the intervention. INTERVENTION: Parents who participated in an educational training received free seats, educational programs were provided to all daycare staff and children, and signs in parking lots informed parents about child restraints. At seven centers, new policies recommended compliance with state restraint laws. Parents at four centers randomly chosen from the seven received financial incentives if observed using booster seats. MAIN OUTCOME MEASURE: The percent of children aged 4-8 riding in booster seats. RESULTS: Pre-test observations of 185 4-8 year olds found 56% riding unrestrained and fewer than 3% riding in booster seats. After the intervention, observation of 146 children found the number riding in booster seats increased to 38% and the number observed without restraints decreased to 26%. Most booster seat use occurred with 4 and 5 year olds. No 7 or 8 year olds rode in booster seats. Changing center policies to recommend compliance with state restraint laws and an offer of financial incentives appeared to have no additional impact. CONCLUSIONS: Booster seat usage among low income families can be increased dramatically, though use decreases with age. Providing free seats accompanied by training may be sufficient without the need for additional intervention.

Immunotherapy of localized, intermediate, and diffuse forms of American cutaneous leishmaniasis.

The clinical efficacy of immunotherapy for localized American cutaneous leishmaniasis with a combination of heat-killed Leishmania mexicana amazonensis promastigotes and viable BCG (bacille Calmette Guérin) has been compared with meglumine antimoniate chemotherapy and with BCG alone in a controlled clinical study in 217 patients. The results in the first two groups were comparable, with greater than 90% clinical cures with an average time of 16-18 w required for healing. The cure rate was considerably lower (42%) and more prolonged in the group receiving BCG alone. Secondary effects were observed in less than 5% of the patients receiving combined immunotherapy or BCG alone. In contrast, 49% of the patients receiving chemotherapy showed side effects. High therapeutic efficacy was also observed using combined immunotherapy in patients with intermediate and diffuse cutaneous leishmaniasis who were previously unresponsive to chemotherapy. Cure or clinical improvement was seen in all 11 patients with intermediate forms of the disease, and marked clinical improvement was observed in 9 of 10 patients with diffuse disease. The results on the efficacy of the combined vaccine in immunotherapy for American cutaneous leishmaniasis provide a strong rationale for studying its effectiveness in prophylactic trials.

Immune response in healthy volunteers vaccinated with killed leishmanial promastigotes plus BCG. I: Skin-test reactivity, T-cell proliferation and interferon-gamma production.

This study reports the results of a vaccine trial established to study the cellular immune responses in vivo (skin-test reactivity) and in vitro (T-cell proliferation and interferon-gamma production) to both leishmanial and mycobacterial antigens following vaccination of healthy volunteers from a leishmaniasis-endemic area with killed leishmanial promastigotes, with or without BCG (Bacille Calmètte-Guerin). Skin tests were performed using purified protein derivative of tuberculin (PPD) and leishmanial antigen in 692 volunteers, and 208 doubly negative subjects (< or = 7 mm induration) were selected to participate in the trial. The study subjects were divided into four vaccine groups: (A) killed promastigotes plus BCG, (B) BCG alone, (C) killed promastigotes alone, and (D) placebo. Three vaccine doses were administered at 6-10-week intervals. The skin-test responses to PPD and leishmanial antigen were reassessed at 4-6- and 12-18-month follow-ups. The results of this trial demonstrated that the combined vaccine, i.e. killed promastigotes of Leishmania plus BCG, results in the stimulation of an immune response to both leishmania and mycobacterial antigens in a high percentage of vaccines (> 85%), manifested either by skin-test conversion, lymphocyte proliferation and/or interferon-gamma production. This was evident after the first dose of vaccine for lymphocyte proliferation and interferon-gamma production and was maintained for a year after the three doses of vaccine. Group B (which received BCG alone), responded as well as group A to PPD but not as well to leishmanial antigen. The reverse was true for group C which received promastigotes alone. Group A attained a 38% leishmanin skin-test conversion at the 4-6-month follow-up, which was associated with double PPD/leishmanial antigen responder status. In contrast, a 35% skin-test conversion was found at the 12-18-month follow-up in group C (promastigotes alone), but this was not associated with responses to PPD. A significant percentage of conversion was observed in the placebo group at the 12-18-month follow-up, both to PPD (58%) and leishmanial (21%) antigens, which suggests either environmental exposure to mycobacterial or leishmanial antigens during the vaccine trial or, more probably, a response to the repeated leishmanial skin tests. Further studies are required to determine whether the presence of proliferative and/or interferon-gamma responses in the absence of a skin-test response are sufficient indicators of potential vaccine success.