Gold nanoparticle-enhanced radiotherapy: Dependence of the macroscopic dose enhancement on the microscopic localization of the nanoparticles within the tumor vasculature.

In gold nanoparticle-enhanced radiotherapy, intravenously administered nanoparticles tend to accumulate in the tumor tissue by means of the so-called permeability and retention effect and upon irradiation with x-rays, the nanoparticles release a secondary electron field that increases the absorbed dose that would otherwise be obtained from the interaction of the x-rays with tissue alone. The concentration of the nanoparticles in the tumor, number of nanoparticles per unit of mass, which determines the total absorbed dose imparted, can be measured via magnetic resonance or computed tomography images, usually with a resolution of several millimeters. Using a tumor vasculature model with a resolution of 500 nm, we show that for a given concentration of nanoparticles, the dose enhancement that occurs upon irradiation with x-rays greatly depends on whether the nanoparticles are confined to the tumor vasculature or have already extravasated into the surrounding tumor tissue. We show that, compared to the reference irradiation with no nanoparticles present in the tumor model, irradiation with the nanoparticles confined to the tumor vasculature, either in the bloodstream or attached to the inner blood vessel walls, results in a two to three-fold increase in the absorbed dose to the whole tumor model, with respect to an irradiation when the nanoparticles have already extravasated into the tumor tissue. Therefore, it is not enough to measure the concentration of the nanoparticles in a tumor, but the location of the nanoparticles within each volume element of a tumor, be it inside the vasculature or the tumor tissue, needs to be determined as well if an accurate estimation of the resultant absorbed dose distribution, a key element in the success of a radiotherapy treatment, is to be made.

Effects of protein and forage source on performance, and splanchnic and mammary net fluxes of nutrients in lactating dairy cows.

This study was conducted to determine if the decreased MP supply predicted by the NRC (2001) when canola meal (CM) substitutes soybean meal (SBM) was supported by direct measurement of net portal absorption of AA or energy-yielding nutrients, plus the impact of the type of forage in CM-based rations. Nine Holstein cows with indwelling catheters in splanchnic blood vessels, 8 also with a ruminal cannula were used to examine the effects of protein source in corn silage-based diets, comparing SBM versus CM, and forage source in CM-based diets, comparing corn versus grass silage. The cows were allocated to a triple 3 × 3 Latin square design with 21-d periods. The 3 experimental diets, formulated to be isoenergetic and isonitrogenous, were based on: 1) SBM and corn silage (SoyCorn); 2) CM and corn silage (CanCorn) and 3) CM and cool-season grass silage (CanGrass). Averages of intake, milk yield and milk composition of the last 3 d of each period were used for statistical analyses. On d 21 of each period, 6 sets of arterial, portal, hepatic and mammary blood samples and 2 ruminal fluid samples were collected. On d 12 of period 2, the protein sources were incubated in nylon bags to determine 16h-ruminal disappearance of DM and N and to obtain 16-h residues. Finally, 5 d after the completion of the Latin square design, the mobile bag technique was used to determine DM and N intestinal disappearance of the 16-h residues of SBM and CM. Pre-planned contrasts were used to compare the effect of the protein source in cows fed corn silage, i.e., SoyCorn versus CanCorn, and the effect of forage in cows fed CM, i.e., CanCorn versus CanGrass. Data of the cow without a rumen canula could not be used because of health problem. In corn silage-based diets, substitution of SBM by CM tended to increase milk (6%) and milk fat (7%) yields. The 8% higher ruminal N disappearance and the 19% decreased MP supply from RUP predicted by NRC (2001) were not supported by the 25% decrease in ruminal ammonia concentration, similar net portal absorption of AA (except 22% higher for Met), and the 14% decrease in urea hepatic removal when CM substituted SBM. Ruminal incubation of CM in nylon bags does not appear suitable for adequate determination of the rumen by-pass of a protein source like CM. Inclusion of grass silage rather than corn silage in CM-based diets tended to increase milk (6%) and increased milk lactose (8%) yields. Neither protein nor forage source resulted in variations of metabolism of energy-yielding nutrients that could explain observed increments in cow performance. The present study indicates no decreased AA availability when CM substitutes SBM. Therefore, substitution of SBM by CM in diets based on corn silage and CM in corn- or grass silage-diets can be used successfully in high producing dairy cows.

Clinical practice guideline of the Spanish society of oral surgery for dental care in patients with oral cancer.

BACKGROUND: Oral cancer is the sixteenth most common malignant neoplasm worldwide, with a high mortality rate, greater than 50% at five years, and high morbidity. The effect of oncological treatment in the oral cavity is broad and has multiple levels, therefore knowing these effects and preventing them is essential for avoiding an increase in the oral pathology related with oncological therapy, maintaining the quality of life of the patient, and improving the efficacy of the treatment itself. MATERIAL AND METHODS: A group of experts belonging to the fields of Dentistry, Maxillofacial Surgery and Oncology of the University of Seville and the Virgen del Rocío University Hospital of Seville in collaboration with the University of Valencia, University of Barcelona, and University of the Basque Country, developed this Clinical Practice Guideline for the proper clinical management of patients diagnosed with oral cancer. The clinical questions were formulated in PICO format. The databases consulted were Medline/PubMed and Embase/Elsevier. The systematic reviews published on the topic were identified on Tripdatabase, Cochrane Library and CRD (Centre for Reviews and Dissemination). The recommendations were prepared based on the GRADE methodology. RESULTS: Various recommendations were defined, derived from the 21 PICO questions, referring to prevention, treatment and care for alterations arising from the pathology of oral cancer itself and its treatment. CONCLUSIONS: The preparation of this clinical practice guideline allows recommendations to be generated based on the scientific evidence available, on dentistry actions in patients with oral cancer and undergoing oncological treatment, which may be of use to the multidisciplinary team treating this type of patient.

[Comprehensive ergonomics program for the reduction of musculoskeletal discomfort in computer user workers]. / Programa integral de ergonomía para la reducción de molestias musculoesqueléticas en trabajadores usuarios de computadora.

INTRODUCTION: Musculoskeletal complaints represent one of the main problems that affect the well-being and work performance of computer users demanding interventions with a comprehensive approach where workers actively participate. OBJECTIVE: To evaluate the effect of a comprehensive ergonomics program to reduce musculoskeletal discomfort and postural overload in computer user workers. METHODS: A quantitative study with an analytical approach and quasi-experimental design with independent groups was carried out during 3-month. 242 people participated, divided into 2 groups of 121 people each, who used a computer for more than 4h a day. The experimental group participated in the ergonomics program that included training, delivery of an informative leaflet on ergonomics in offices, adaptation of the workstation and a stretching exercise program. The control group received training and only once the information leaflet. A baseline measurement of musculoskeletal discomfort and postural overload was performed in both groups. At the end of 3months, a second data collection was carried out and compared with the baseline data. RESULTS: The ergonomics program significantly reduced the intensity of musculoskeletal discomfort in the different neck, shoulder and back segments (P<.05), but did not reduce the frequency of discomfort (P>.05). Furthermore, the program significantly reduced the level of postural overload (P<.05). CONCLUSION: The comprehensive ergonomics program was effective in reducing the intensity of musculoskeletal complaints and postural overload, but did not significantly reduce the frequency of musculoskeletal complaints.

Aryl hydrocarbon receptor interacting protein (AIP) significantly influences prognosis of pancreatic carcinoma.

INTRODUCTION: Aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor. Aryl hydrocarbon receptor interacting protein (AIP) in one of AHR ligands. The aim of this study is to analyze the prognostic influence of AIP in pancreatic carcinoma. MATERIAL AND METHODS: Retrospective case series with immunohistochemical analysis of AIP. We have estimated a multivariate Cox's model for the outcome (progression free and overall survival). RESULTS: 204 patients were included in the study. As expected prognosis was poor and 67.8% died of disease. As for AIP 9.8% of the cases showed nuclear staining of the epithelial tumor cells and 59.4% a cytoplasmic one. Stroma was stained in 53.1% of the cases. Univariate survival analysis revealed a significantly worse prognosis of patients with cytoplasmic AIP expression (stroma and epithelium), but nuclear expression was associated to a better prognosis. In the multivariate analysis stromal AIP expression was an independent prognosticator of progression free survival, together with pT stage, histological grade and history of diabetes. DISCUSSION: AIP Is a conserved cochaperone protein binding to many proteins. AIP has been proposed as a potential tumor suppressor gene. To date, no study has analyzed the immunohistochemical expression of AIP in pancreatic carcinoma. Our results indicate that both epithelial and stromal cytoplasmic expression of AIP is associated to bad prognosis, while nuclear translocation seems to improve prognosis. CONCLUSION: Although we must deepen into the complex signaling pathways underlying this potential association, our results open a way to inhibiting AHR as a potential target against pancreatic carcinoma.

Recommendations for the prevention and diagnosis of asthma in children: Evidence from international guidelines adapted for Mexico.

BACKGROUND: With the availability of high-quality asthma guidelines worldwide, one possible approach of developing a valid guideline, without re-working the evidence, already analysed by major guidelines, is the ADAPTE approach, as was used for the development of National Guidelines on asthma. METHODS: The guidelines development group (GDG) covered a broad range of experts from medical specialities, primary care physicians and methodologists. The core group of the GDG searched the literature for asthma guidelines 2005 onward, and analysed the 11 best guidelines with AGREE-II to select three mother guidelines. Key clinical questions were formulated covering each step of the asthma management. RESULTS: The selected mother guidelines are British Thoracic Society (BTS), GINA and GEMA 2015. Responses to the questions were formulated according to the evidence in the mother guidelines. Recommendations or suggestions were made for asthma treatment in Mexico by the core group, and adjusted during several rounds of a Delphi process, taking into account: 1. Evidence; 2. Safety; 3. Cost; 4. Patient preference - all these set against the background of the local reality. Here the detailed analysis of the evidence present in BTS/GINA/GEMA sections on prevention and diagnosis in paediatric asthma are presented for three age-groups: children with asthma ≤5 years, 6-11 years and ≥12 years. CONCLUSIONS: For the prevention and diagnosis sections, applying the AGREE-II method is useful to develop a scientifically-sustained document, adjusted to the local reality per country, as is the Mexican Guideline on Asthma.

Patient-derived xenografts effectively capture responses to oncology therapy in a heterogeneous cohort of patients with solid tumors.

BACKGROUND: While patient-derived xenografts (PDXs) offer a powerful modality for translational cancer research, a precise evaluation of how accurately patient responses correlate with matching PDXs in a large, heterogeneous population is needed for assessing the utility of this platform for preclinical drug-testing and personalized patient cancer treatment. PATIENTS AND METHODS: Tumors obtained from surgical or biopsy procedures from 237 cancer patients with a variety of solid tumors were implanted into immunodeficient mice and whole-exome sequencing was carried out. For 92 patients, responses to anticancer therapies were compared with that of their corresponding PDX models. RESULTS: We compared whole-exome sequencing of 237 PDX models with equivalent information in The Cancer Genome Atlas database, demonstrating that tumorgrafts faithfully conserve genetic patterns of the primary tumors. We next screened PDXs established for 92 patients with various solid cancers against the same 129 treatments that were administered clinically and correlated patient outcomes with the responses in corresponding models. Our analysis demonstrates that PDXs accurately replicate patients' clinical outcomes, even as patients undergo several additional cycles of therapy over time, indicating the capacity of these models to correctly guide an oncologist to treatments that are most likely to be of clinical benefit. CONCLUSIONS: Integration of PDX models as a preclinical platform for assessment of drug efficacy may allow a higher success-rate in critical end points of clinical benefit.

Risk of second malignant neoplasms in women and girls with germ cell tumors.

Background: While an elevated risk of second malignant neoplasms (SMNs) has been observed in men treated for germ cell tumors (GCTs), risk of SMNs have not been quantified in adult women or in girls treated for GCTs. Patients and methods: One-year survivors of primary GCTs diagnosed between January 1980 and December 2012 were identified from Surveillance, Epidemiology, and End Results (SEER 9) registries. Risk of SMNs was calculated using SEER*Stat. Results: Among 1507 patients, a total of 47 SMNs were identified. The overall risk of SMNs was not elevated in females overall or in females treated for GCT during adulthood although SMN sites (pancreas, soft tissue, bladder, kidney, and thyroid) and trends were comparable with those in men. There were too few childhood GCT cases with SMNs for further analysis. Conclusions: Unlike men, women treated for GCTs did not have a statistically significant elevated risk of SMNs [standardized incidence ratio = 1.11; 95% confidence interval (CI) = 0.81-1.47]. The fact that SMNs in women occur in sites similar to those observed in men indicate that long-term follow-up of a larger cohort of females treated for GCT is warranted.

Response to chemotherapy estimates by FDG PET is an important prognostic factor in patients with Ewing sarcoma.

BACKGROUND: Response to chemotherapy is a prognostic factor in patients with Ewing sarcoma (ES); the role of FDG PET to predict response in these patients has not been thoroughly investigated. We evaluated the diagnostic accuracy and the potential of FDG PET to predict response to chemotherapy (CHT). MATERIALS AND METHODS: We analyzed data of 50 patients with ES (median age 12.6 years). All patients were treated with neoadjuvant CHT, and underwent surgery for local control. All patients had (18)F-FDG PET/CT at diagnosis and after induction CHT, prior to local control. We compared response assessed by histopathology with FDG PET using standard uptake values (SUVs). RESULTS: Median SUV at diagnosis (SUV I) was 5 (range 1.2-17), and median SUV after neoadjuvant chemotherapy (SUV II) was 1.8 (range 0-8.4). Median SUV II/I ratio was 0.3 (range 0-1). SUV at diagnosis was significantly lower in patients with good histological response than in patients with poor histological response (median 3.8 vs. 7.2, p 0.02). We found a significant correlation between SUV II and outcome; the positive predictive value of an SUV II ≤ 2.5 for favorable response was 84.21 %, and the median SUV II was significantly higher in patients with disease progression (2.3 vs. 1.6, p = 0.04). In multivariate analysis, necrosis and SUV II were significant predictors of outcome. CONCLUSIONS: (18)F-FDG PET demonstrates high diagnostic accuracy for response to initial chemotherapy in patients with ES and it correlates with outcome. The role of FDG PET in predicting response and outcome should be further investigated.

Abomasal amino acid infusion in postpartum dairy cows: Effect on whole-body, splanchnic, and mammary glucose metabolism.

Nine Holstein cows fitted with rumen cannulas and indwelling catheters in splanchnic blood vessels were used to study the effects of supplementing AA on milk lactose secretion, whole-body rate of appearance (WB-Ra) of glucose, and tissue metabolism of glucose, lactate, glycerol, and ß-OH-butyrate (BHBA) in postpartum dairy cows according to a generalized randomized incomplete block design with repeated measures in time. At calving, cows were blocked according to parity (second and third or greater) and were allocated to 2 treatments: abomasal infusion of water (n=4) or abomasal infusion of free AA with casein profile (AA-CN; n=5) in addition to the same basal diet. The AA-CN infusion started with half the maximal dose at 1 d in milk (DIM) and then steadily decreased from 791 to 226 g/d from DIM 2 to 29 to cover the estimated essential AA deficit. On DIM 5, 15, and 29, D[6,6-(2)H2]-glucose (23.7 mmol/h) was infused into a jugular vein for 5h, and 6 blood samples were taken from arterial, portal, hepatic, and mammary sources at 45-min intervals, starting 1h after the initiation of the D[6,6-(2)H2]glucose infusion. Trans-organ fluxes were calculated as veno-arterial differences times plasma flow (splanchnic: downstream dilution of deacetylated para-aminohippurate; mammary: Fick principle using Phe+Tyr). Energy-corrected milk and lactose yields increased on average with AA-CN by 6.4 kg/d and 353 g/d, respectively, with no DIM × treatment interaction. Despite increased AA supply and increased demand for lactose secretion with AA-CN, net hepatic release of glucose remained unchanged, but WB-Ra of glucose tended to increase with AA-CN. Portal true flux of glucose increased with AA-CN and represented, on average, 17% of WB-Ra. Splanchnic true flux of glucose was unaltered by treatments and was numerically equivalent to WB-Ra, averaging 729 and 741 mmol/h, respectively. Mammary glucose utilization increased with AA-CN infusion, averaging 78% of WB-Ra, and increased gradually as lactation advanced. Net portal, hepatic, splanchnic, and mammary fluxes of lactate, glycerol, and BHBA were not affected by AA infusion. Increasing the supply of AA in postpartum dairy cows elevated the WB-Ra of glucose without affecting the true liver glucose release. The greater WB-Ra of glucose with abomasal AA infusion seemed to originate mainly from greater true portal-drained viscera release of glucose. Glucose utilization by the portal-drained viscera was unaffected by abomasal AA infusion, but the exact mechanism behind the greater true portal glucose release could not be assessed in the current study. The increased mammary glucose uptake was in line with the increased milk lactose yield. In early postpartum lactation, the demand for AA seems to be so high that even with increased AA supply, cows have metabolic priorities for AA other than hepatic gluconeogenesis.

Abomasal amino acid infusion in postpartum dairy cows: Effect on whole-body, splanchnic, and mammary amino acid metabolism.

Nine Holstein cows with rumen cannulas and indwelling catheters in splanchnic blood vessels were used in a generalized randomized incomplete block design with repeated measures to study the effect of increased early postpartum AA supply on splanchnic and mammary AA metabolism. At calving, cows were blocked according to parity (second and third or greater) and allocated to 2 treatments: abomasal infusion of water (CTRL; n=4) or free AA with casein profile (AA-CN; n=5) in addition to a basal diet. The AA-CN infusion started with half of the maximal dose at the calving day (1 d in milk; DIM) and then steadily decreased from 791 to 226 g/d until 29 DIM. On 5, 15, and 29 DIM, 6 sample sets of arterial, portal, hepatic, and mammary blood were taken at 45-min intervals. Over the whole period, increasing AA supply increased milk (+7.8 ± 1.3 kg/d) and milk protein yields (+220 ± 65 g/d) substantially. The increased milk yield was not supported by greater dry matter intake (DMI) as, overall, DMI decreased with AA-CN (-1.6 ± 0.6 kg/d). Arterial concentrations of essential AA were greater for AA-CN compared with CTRL. The net portal-drained viscera (PDV) release of His, Met, and Phe was greater for AA-CN compared with CTRL, and the net PDV recovery of these infused AA ranged from 72 to 102% once changes in DMI were accounted for. The hepatic removal of these AA was increased equivalently to the increased net PDV release, resulting in an unaltered net splanchnic release. The net PDV release of Ile, Leu, Val, and Lys tended to be greater for AA-CN, and the net PDV recovery of these infused AA ranged from 69 to 73%, indicating increased PDV metabolism with AA-CN. The fractional hepatic removal of these AA did not differ from zero and was unaffected by the increased supply. Consequently, the splanchnic release of these AA was approximately equivalent to their net PDV release for both CTRL and AA-CN. Overall, greater early postpartum AA supply increased milk and milk protein yields substantially based on increased mammary AA uptake. The PDV metabolism of branched-chain AA and Lys were increased, whereas it seemed to be unaffected for other essential AA when the intestinal AA supply was increased. On a net basis, the liver removed more group 1 AA (His, Met, Phe, and Trp) for anabolism and catabolism when the early postpartum AA supply was increased. Thus, increasing the postpartum AA supply increased splanchnic and mammary consumption of AA; hence, the protein deficiency persisted.

Recurrent osteosarcoma with a single pulmonary metastasis: a multi-institutional review.

BACKGROUND: Late relapse and solitary lesion are positive prognostic factors in recurrent osteosarcoma. METHODS: We reviewed the records of 39 patients treated at three major centres for recurrent osteosarcoma with a single pulmonary metastasis more than 1 year after diagnosis. We analysed their outcomes with respect to clinical factors and treatment with chemotherapy. RESULTS: Median age at diagnosis was 14.6 years. Relapse occurred at a median of 2.5 years (range, 1.2-8.2 years) after initial diagnosis. At relapse, all patients were treated by metastasectomy; 12 (31%) patients also received chemotherapy. There was no difference in time to recurrence or nodule size between the patients who received or did not receive chemotherapy at relapse. Sixteen patients had no subsequent recurrence, 13 of whom survive without evidence of disease. The 5-year and 10-year estimates of post-relapse event-free survival (PREFS) were 33.0±7.5% and 33.0±9.6%, respectively, and of post-relapse survival (PRS) 56.8±8.6% and 53.0±11.0%, respectively. There was a trend for nodules <1.5 cm to correlate positively with PREFS (P=0.070) but not PRS (P=0.49). Chemotherapy at first relapse was not associated with PREFS or PRS. CONCLUSION: Approximately half of the patients with recurrent osteosarcoma presenting as a single pulmonary metastasis more than 1 year after diagnosis were long-term survivors. Metastasectomy was the primary treatment; chemotherapy did not add benefit.

Validación de contenido de un cuestionario de satisfacción usuaria con la atención abierta en un policlínico ambulatorio de otorrinolaringología / User satisfaction with open attention on an outpatient otolaryngology clinic

Introducción: La satisfacción de los pacientes con respecto a la atención en salud que ofrecen las instituciones es relevante. Objetivo: Realizar validación de contenido de un cuestionario que permita analizar la opinión de un grupo de pacientes consultantes a un policlínico de otorrinolaringología respecto al proceso de atención médica. Material y método: Estudio de corte transversal. Se validó un instrumento mediante el método de Lawshe. Se crearon 7 reactivos, con un total de 83 preguntas para ser sometidas a análisis. Resultados: Se validaron 25 preguntas que conformaron el cuestionario final, con una razón de validez de contenido de 0,75 o más. Fueron encuestados 120 pacientes entre 30 y 60 años, correspondientes a 83 mujeres (69,1%). Del total, 75 pacientes (62,5%) acudían por primera vez a consulta, y 36 pacientes (30%) habían sido operados. Más del 94% de las pacientes refirió que el médico les explicó claramente su diagnóstico y dio con claridad las indicaciones para tratar su enfermedad. El 90,8% refirió que le parecía adecuada la presencia de estudiantes, internos o becados en el box. El 95,8% de los pacientes refirió sentirse satisfecho con la atención recibida. Conclusiones: Las cifras generales de satisfacción en la población estudiada son positivas. El cuestionario podría ser de utilidad para medir satisfacción usuaria. Es necesario seguir monitorizando la percepción de los pacientes sobre la atención médica.

Introduction: Patient satisfaction with respect to health care institutions is relevant. Aim: To perform content validation a questionnaire that allows to analyze the opinion of a group of patients attending an ENT clinic regarding on the process of care in this place. Material and method: Cross sectional study. The instrument was validated through Lawshe method. Seven reagents were created, with a total of 83 questions to be subjected to analysis. Results: We validated 25 questions that made the final questionnaire, with a content validity ratio of 0.75 or more. We surveyed 120 patients between 30 and 60 years, being 83 women (69.1%) and 37 men (30.9%). Of the total, 75 patients (62.5%) came to first consultation, and 36 patients (30%) had been operated. More than 94% of the patients said that the doctor clearly explained his diagnosis and clearly gave indications to treat their condition. 90.8% said that it seemed appropriate the presence of students, interns and residents or fellows in the consultation box. 95.8% of patients reported being satisfied with the care provided. Conclusions: The overall high level of satisfaction in the study population is positive. The questionnaire could be useful for measuring user satisfaction. It is necessary to continue to monitor patients' perception of care.

[Effect on zebrafish neurodevelopment and neuroprotection of a polyphenolic extract olive seeds]. / Efecto sobre el neurodesarrollo y neuroprotección en pez cebra de un extracto polifenólico de huesos de aceituna.

OBJECTIVE: To determine the effect of a polyphenolic extract from olive pit on the development of the nervous system as well as its effect on pain induced by the neurotoxin kainic acid, taking the zebrafish as the animal model. MATERIAL AND METHODS: We analyse the effect of the extract at the maximum tolerated dose (100 mg/ml of polyphenols) on the cholinergic activity in zebrafish larvae (72 hours post-fertilization). Only fecundated eggs with no abnormalities are used. 6 eggs/bowl are incubated in a 24 bowls microplate in 2 ml of water with DMSO (0.1%) at 26 ± 1º C: a) neurodevelopment: water (control) and 100 mg/ml of extract, as an essay; b) neuroprotection: water and kainic acid (100 µM) (control) and 100 mg/ml of extract (essay). All incubations are in triplicate. After 72 h, incubations are examined and checked for any abnormalities. Larvae are homogenized and acetyl cholinesterase activity and protein concentration in supernatants is quantified. RESULTS: The quantity of protein and the morphologic appreciation is similar in all the essays, showing a standard development. Acetyl cholinesterase in fish larvae, with the polyphenolic extract is 162.2% (SD 44.2) compared to controls (100% of activity) (p < 0.01). Fish larvae treated with kainic acid and polyphenolic acid show 140.1% (SD 22.0) of activity, compared to those only incubated with the neurotoxin (100%) (p < 0.05). CONCLUSION: Polyphenols extracted from olive pit produce an increase in the cholinergic activity during the larvae neurodevelopment in the zebrafish as well as protection against the neurotoxin kainic acid.

Objetivo: Determinar el efecto de un extracto polifenólico de hueso de oliva en el desarrollo del sistema nervioso y frente al daño inducido mediante la neurotoxina ácido kaínico, utilizando como modelo animal el pez cebra. Material y métodos: Se analiza el efecto del extracto a la máxima dosis tolerada (100 mg/ml de polifenoles) sobre la actividad colinérgica en larvas de pez cebra (72 horas post-fertilización). Se utilizan únicamente huevos fecundados sin anomalías. Se incuban 6 huevos/pocillo en microplaca de 24 pocillos en 2 ml de agua con DMSO (0,1%) a 26 ± 1º C: a) neurodesarrollo: agua (control) y con 100 mg/ml de extracto, como ensayo; b) neuroprotección: agua y ácido kaínico (100 M) (control) y con 100 mg/ml de extracto (ensayo). Todas las incubaciones por triplicado. A las 72 h se examinan y verifica ausencia de anomalías. Las larvas se homogeneizan y en los sobrenadantes se cuantifica actividad acetilnolinesterasa y concentración proteínas. Resultados: La cantidad de proteína y apreciación morfológica es análoga en todos los ensayos, indicando mismo desarrollo. La acetilcolinesterasa en las larvas de pez, con el extracto polifenólico es del 162,2%(SD 44,2) respecto a controles (100% de actividad) (p < 0,01). Las larvas de pez tratadas con ácido kaínico y extracto polifenólico presentan el 140,1% (SD 22,0) de actividad, respecto a las incubadas únicamente con la neurotoxina (100%) (p < 0,05). Conclusión: Los polifenoles extraídos de los huesos de aceituna producen incremento de actividad colinérgica durante el neurodesarrollo larvario en el pez cebra y protección frente a la neurotoxina ácido kaínico.

[Anti-inflammatory activity of olive seed polyphenolic extract in the THP1-XBLUE-CD14 human monocytes cell line]. / Actividad antiinflamatoria de un extracto polifenólico de hueso de olivas en la línea celular de monocitos humanos THP1-XBLUE-CD14.

UNLABELLED: The aim of this study was to assess the anti-inflammatory activity of a polyphenolic extract from olive pits. MATERIAL AND METHODS: The THP1-XBlue-CD14 (invivogen) cellular line, 80,000 cells/well, was incubated and inflammation (activation of NF-kb) was produced with 0.1 mg/mL of LPS (lipopolysaccharide from E. coli) for 24 hours. We assessed the presence of the extract (10 and 50 mg/L, biologically safe concentrations) for 2 hours at 37º C, before (preventive effect) and after (therapeutic effect) the proinflammatory activation, and the activity of alkaline phosphatase, which is expressed under the control of the NF-kb transcriptional factor, was quantified by colorimetry. The percentage of activity of NF-kb as preventive effect and therapeutic effect was assessed by comparing it to control cultures of cells with LPS and without extract, which are considered 100% of NF-kb. RESULTS: The preventive anti-inflammatory capacity of the extract at 50 mg/L was 25.5% (95% CI: 16.8-34.2) and the therapeutic effect 34.9% (95% CI: 25.3-44.4) for the same concentration, without any significant activity at 10 mg/L. CONCLUSION: An activity of polyphenols extracted from olive pits is shown, both in preventing inflammation and therapeutically eliminating inflammation through inhibition of NF-kB factor previously activated by LPS at concentrations of 50 mg/L of polyphenols, which previously haven been shown to be safe.

El objetivo de este trabajo es evaluar la actividad antiinflamatoria de un extracto de naturaleza polifenólica de huesos de oliva. MATERIAL Y MÉTODOS: Se incubó la línea celular THP1- XBlue-CD14 (invivogen), 80.000 células/pocillo, provocando inflamación (activación de NF-kb) mediante 0.1 µg/ml LPS (lipopolisacárido de E. coli) durante 24 horas. Se evaluó la presencia del extracto (10 y 50 mg/l, concentraciones bioseguras) durante 2 horas a 37 ºC, previa (efecto preventivo) y posterior a la activación proinflamatoria (efecto terapéutico) y se cuantificó colorimétricamente la actividad de fosfatasa alcalina, que se expresa bajo el control del promotor del factor transcripcional de NF-kb. Se evalúa el % actividad de NF-kb en efecto preventivo y terapéutico respecto a cultivos control de células con LPS y sin extracto añadido, que se consideran 100% de NF-kb. RESULTADOS: La capacidad antiinflamatoria preventiva del extracto a 50 mg/l es del 25,5% (IC 95% 16,8-34,2) y el efecto terapéutico del 34,9% (IC 95% 25,3-44,4) para la misma concentración, no presentando actividad significativa a 10 mg/l. CONCLUSIÓN: Se muestra una actividad de los polifenoles extraídos de los huesos de aceitunas, tanto preventivo de la inflamación como terapéutico de eliminación de la inflamación a través de la inhibición del factor NF-kB previamente activado por LPS a concentraciones de 50 mg/l de polifenoles que previamente se han mostrado seguras.

[Assessment of cytotoxicity and biosafety of polyphenolic extracts from olive pits]. / Evaluación de la citotoxicidad y bioseguridad de un extracto de polifenoles de huesos de aceitunas.

UNLABELLED: The olive tree is a source of bioactive compounds, both its fruit and its by-products. Some of its compounds have shown health benefits, being objective of this work the evaluation of biosafety in-vitro and in vivo of extracts of olive stones rich in polyphenols. MATERIAL AND METHODS: He has been evaluated for cytotoxicity by addition of lyophilized extract dissolved in PBS(0-400 mg/l) to a culture of the cell line THP1-XBlue- CD14 and evaluation of cell viability by the reaction of reduction of resazurin by living cells. Biosecurity has been evaluated in zebrafish, incubating eggs fertilized in 0 to 100 mg/l extract for 24 to 72 hours and measuring parameters: a) lethal (dead embryos, coagulated eggs), b) sublethal (spontaneous movements, pigmentation, edemas) and c) teratogenic (malformations, retardation development). RESULTS: Cytotoxicity (toxic effect when less than 75% viability) extract bones of olive in the cell line THP1- XBlue-CD14, is in concentrations higher than 50 mg/l, calculating a LD50 (dose lethality 50) more than 800 mg/l. The biosafety of zebrafish embryos exposed to concentrations of extract from 0-100 mg/l showed total viability at 24, 48 and 72 hours post fertilization (hpf), not observed mortality or appreciated embryos with sublethal effects, teratogenic, or advancement or delay in hatching. It can be concluded that the bones of olive extract is highly biosecured until at least 100 mg/l concentrations.

El olivo constituye una fuente de compuestos bioactivos, tanto en su fruto, como en sus subproductos. Algunos de sus compuestos han mostrado beneficios para la salud, siendo objetivo de este trabajo la evaluación de la bioseguridad in vitro e in vivo de extractos de huesos de aceituna ricos en polifenoles. Material y métodos: Se ha evaluado la citotoxicidad mediante adición de extracto de hueso de olivas disuelto en PBS(0-400 mg/l) a un cultivo de la línea celular THP1- XBlue-CD14 y evaluación de la viabilidad celular mediante la reacción de reducción de la resazurina por las células vivas. La bioseguridad se ha evaluado en pez cebra, incubando huevos fecundados en extracto de 0 a 100 mg/l durante 24 a 72 horas y midiendo los parámetros: a) letales (embriones muertos, huevos coagulados), b) subletales (movimientos espontáneos, pigmentación, edemas) y c) teratogénicos (malformaciones, retraso desarrollo). Resultados: La citotoxicidad (efecto tóxico cuando viabilidad inferior al 75%) del extracto de huesos de oliva en la línea celular THP1-XBlue-CD14, está en concentraciones superiores a 50 mg/l de extracto (viabilidad 77,5%), calculando una LD50 (dosis de letalidad 50%) superior a 800 mg/l. La bioseguridad in vivo con los embriones de pez cebra expuestos a concentraciones de extracto de 0- 100 mg/l mostró total viabilidad a 24, 48 y 72 horas post fecundación (hpf), no observándose mortalidad ni se apreciaron embriones con efectos subletales, teratógenos, ni adelanto o retraso en la eclosión. Se puede concluir que el extracto de huesos de olivas es altamente bioseguro hasta al menos concentraciones de 100 mg/l.

Exome sequencing of pleuropulmonary blastoma reveals frequent biallelic loss of TP53 and two hits in DICER1 resulting in retention of 5p-derived miRNA hairpin loop sequences.

Pleuropulmonary blastoma is a rare childhood malignancy of lung mesenchymal cells that can remain dormant as epithelial cysts or progress to high-grade sarcoma. Predisposing germline loss-of-function DICER1 variants have been described. We sought to uncover additional contributors through whole exome sequencing of 15 tumor/normal pairs, followed by targeted resequencing, miRNA analysis and immunohistochemical analysis of additional tumors. In addition to frequent biallelic loss  of TP53 and mutations of NRAS or BRAF in some cases, each case had compound disruption of DICER1: a germline (12 cases) or somatic (3 cases) loss-of-function variant plus a somatic missense mutation in the RNase IIIb domain. 5p-Derived microRNA (miRNA) transcripts retained abnormal precursor miRNA loop sequences normally removed by DICER1. This work both defines a genetic interaction landscape with DICER1 mutation and provides evidence for alteration in miRNA transcripts as a consequence of DICER1 disruption in cancer.

Incidence of herpes zoster in patients with altered immune function.

PURPOSE: To estimate the incidence of herpes zoster (HZ) and rates of post-zoster pain in both the total study population and separately in patients with selected conditions/treatments associated with altered immune function. METHODS: The health administrative claims databases for commercially insured, Medicare, and Medicaid populations, together accounting for approximately 51 million insured individuals, were analyzed between 2005 and 2009 in a retrospective cohort study. Incidence of HZ episodes per 1,000 person-years (PY) was estimated in all study populations as well as within nine potentially immune-altering conditions. Among patients with HZ, the 6-month rate of persistent post-zoster pain was estimated. RESULTS: Analysis of 90.2 million PY at risk revealed that the incidence of HZ in the total study population was 4.82/1,000 PY. The incidence of HZ was highest among patients with bone marrow or stem cell transplant (43.03 %) followed by solid organ transplant, human immunodeficiency virus infection, and systemic lupus erythematosus [95 % confidence interval (CI) 15.19-17.41 %]. HZ incidence rates were higher among persons on immunosuppressants/chemotherapy than among non-users. In the total study population, HZ incidence increased with age (18-49 years: 3.37/1,000 PY; 65+ years: 8.43/1,000 PY; P < 0.01) and female gender (incidence ratio vs. male 1.39, 95 % CI 1.38-1.40 %). The 6-month rate of persistent post-zoster pain was 4.29 % (95 % CI 4.22-4.36 %), which was higher in patients with the selected conditions. CONCLUSIONS: Despite providing a relatively small fraction of overall HZ cases, persons with immune function-altering conditions make a large contribution to the societal healthcare burden because they have a higher risk of developing HZ and persistent post-zoster pain. These risk factors should be considered in HZ prevention efforts.

Effect of amino acid or casein supply on whole-body, splanchnic, and mammary glucose kinetics in lactating dairy cows.

This study was conducted to establish how AA supplied in a free form or as protein (casein, CN) affect the whole-body rate of appearance (WB Ra) of glucose, splanchnic and mammary glucose kinetics, and milk lactose secretion in lactating dairy cows. Five Holstein cows fitted with a rumen cannula and permanent indwelling catheters in the abomasum, portal, hepatic, and mesenteric veins, and one mesenteric artery, were used in a Youden square with 4 periods of 14 d each. Cows were fed a hay-based diet providing 100 and 70% of their net energy and metabolizable protein (MP) requirements, respectively. Treatments consisted of abomasal infusions of water (70% of MP requirements: control, Con), free AA (95% of MP requirements: AA1; and 120% of MP requirements: AA2), or CN (95% of MP requirements: CN1). The free AA mixture had the same profile as CN. On d 14 of each period, [6,6-(2)H(2)]glucose (25.8 mmol/h) was infused into a jugular vein, and blood samples (n=8) were taken over 4h from arterial, portal, hepatic, and mammary sources to measure glucose enrichment and concentration. Splanchnic and mammary plasma flows were determined by downstream dilution of para-aminohippurate and with the Fick principle, respectively. The last 6 milkings of each period were weighed and sampled to measure the yields of milk and components. The AA1 and CN1 treatments were not different for any of the measured parameters. Supplying AA linearly increased glucose WB Ra (AA2 vs. Con: +151 mmol/h) and liver net flux (+149 mmol/h). Utilization of glucose from the plasma compartment by the portal-drained viscera and liver and true portal absorption were not affected by AA supply. From these observations, we suggest that the increased WB Ra was due to increased net hepatic production. The AA from the infusion, in excess of that used to cover the increase in milk protein, were converted to glucose with an apparent efficiency close to 100% of maximum theoretical efficiency. Milk and lactose yields increased linearly with infusions of AA, by 14 and 16% with AA2 treatment, respectively. However, mammary glucose uptake was not significantly altered by AA infusions; this suggests that the mammary gland exerts active control on the uptake and utilization of glucose. For all treatments, the sum of true portal glucose absorption and true hepatic glucose production contributed more than 99% of WB Ra in the lactating cow; this would suggest that renal glucose synthesis makes only a small contribution to WB Ra under these conditions.

Comparative analysis of the early transcriptome of Brucella abortus--infected monocyte-derived macrophages from cattle naturally resistant or susceptible to brucellosis.

Brucellosis is a worldwide zoonotic infectious disease that has a significant economic impact on animal production and human public health. We characterized the gene expression profile of B. abortus-infected monocyte-derived macrophages (MDMs) from naïve cattle naturally resistant (R) or susceptible (S) to brucellosis using a cDNA microarray technology. Our data indicate that (1) B. abortus induced a slightly increased genome activation in R MDMs and a down-regulated transcriptome in S MDMs, during the onset of infection, (2) R MDMs had the ability to mount a type 1 immune response against B. abortus infection which was impaired in S cells, and (3) the host cell activity was not altered after 12 h post-B. abortus infection in R MDMs while the cell cycle was largely arrested in infected S MDMs at 12 h p.i. These results contribute to an improved understanding of how host responses may be manipulated to prevent infection by brucellae.