Digital image analysis workflows for evaluation of cell behavior and tumor microenvironment to aid therapeutic assessment in high-risk neuroblastoma.

Digital pathology and artificial intelligence are promising emerging tools in precision oncology as they provide more robust and reproducible analysis of histologic, morphologic and topologic characteristics of tumor cells and the surrounding microenvironment. This study aims to develop digital image analysis workflows for therapeutic assessment in preclinical in vivo models. For this purpose, we generated pipelines that enable automatic detection and quantification of vitronectin and αvß3 in heterotopic high-risk neuroblastoma xenografts, demonstrating that digital analysis workflows can be used to provide robust detection of vitronectin secretion and αvß3 expression by malignant neuroblasts and to evaluate the possibility of combining traditional chemotherapy (etoposide) with extracellular matrix-targeted therapies (cilengitide). Digital image analysis added evidence for the relevance of territorial vitronectin as a therapeutic target in neuroblastoma, since its expression is modified after treatment, with a mean percentage of 60.44% in combined therapy tumors vs 45.08% in control ones. In addition, the present study revealed the efficacy of cilengitide for reducing αvß3 expression, with a mean αvß3 positivity of 34.17% in cilengitide treated material vs 66.14% in control and with less tumor growth when combined with etoposide, with a final mean volume of 0.04 cm3 in combined therapy vs 1.45 cm3 in control. The results of this work highlight the importance of extracellular matrix-focused therapies in preclinical studies to improve therapeutic assessment for high-risk neuroblastoma patients.

Assessment of reclaimed agro-wastewater polluted with insecticide residues for irrigation of growing lettuce (Lactuca sativa L) using solar photocatalytic technology.

Scientific literature is full of works studying the removal of different pollutants from water through different Advanced Oxidation Processes (AOPs). Many of them only suggest it is reused for agricultural purposes or for small crops in pots. This study is based on the reuse of reclaimed agricultural wastewater contaminated with four insecticides (chlorantraniliprole, imidacloprid, pirimicarb and thiamethoxam) for growing lettuce in field conditions. First, solar photocatalysis with TiO2/Na2S2O8 was used on a pilot plant in a sunny area (Murcia, SE of Spain) as an environmentally friendly technology to remove insecticide residues and their main reaction intermediates from contaminated water. The necessary fluence (H, kJ m-2) to accomplish 90% removal (H90) ranged from 0.12 to 1212 kJ m-2 for pirimicarb and chlorantraniliprole, respectively. Only six (derived from imidacloprid, pirimicarb and thiametoxam) of 18 transformation intermediate products studied were detected in reclaimed water during the photoperiod (2000 kJ m-2 of accumulated UVA radiation) although all of them were totally photodegraded after a fluence of 1250 kJ m-2. Secondly, reclaimed agro-wastewater was used to irrigate two lettuce crops grown under greenhouse conditions and under agricultural field conditions. In no cases, insecticide residues nor their TIPs were noticed above their respective LOQs (limits of quantification) in soil and lettuce samples (between 0.03 and 0.04 µg kg-1 for pirimicarb and 2.49 and 2.23 µg kg-1 for thiamethoxam, respectively) when they were irrigated with reclaimed water, while residues of the four insecticides and some of their intermediates were found in soil and lettuce by the end of cultivation when they were irrigated with non-reclaimed contaminated water. According to the results, this technology can be applied in a sustainable way, mainly in areas with water scarcity and high solar radiation, contributing to water utilisation in drought areas and the use of renewable energy.

Evaluación de las habilidades no técnicas en cirugía / Assessment of non-technical skills in surgery

Resumen Tradicionalmente, se ha asociado a la falta de habilidades y competencias técnicas con los eventos adversos en el área quirúrgica. Sin embargo, se ha visto que un gran porcentaje de estos eventos se relacionan directamente con déficits en las habilidades no técnicas (HNT) del equipo quirúrgico. Por lo tanto, la evaluación de éstas se ha vuelto un tópico relevante para minimizar errores y fortalecer el desempeño en el escenario operatorio. Múltiples instrumentos han sido desarrollados con este fin, tanto para analizar el funcionamiento del equipo completo, como de cada subgrupo que lo conforma. El presente trabajo pretende resumir los principales aspectos de los distintos métodos que han sido desarrollados en los últimos años para la evaluación de HNT en el área quirúrgica.

Traditionally, the lack of skills and technical competencies has been associated with adverse events in the surgical area. However, it has been seen that a large percentage of these events are directly related to deficits in the non-technical skills (NTS) of the surgical team. Therefore, the evaluation of these has become a relevant topic to minimize errors and strengthen performance in the operating room. Multiple instruments have been developed for this purpose, both to analyze the operation of the complete equipment, and of each subgroup that comprises it. This article aims to summarize the key aspects of the methods that have been developed in recent years for the evaluation of NTS in the surgical area.

Assessment of the thrombin generation assay in haemophilia: comparative study between fresh and frozen platelet-rich plasma.

The severity of haemophilia A has traditionally been classified by the dosage of factor VIII (FVIII) by one-step coagulation tests. However, an homogeneous group of patients with similar FVIII levels show clinical heterogeneity and 10-15% of the patients classified as severe haemophilia do not have a severe bleeding phenotype. Traditional tests used for measuring FVIII are not capable of detecting other prohaemorrhagic or prothrombotic factors. Global tests as the thrombin generation assay (TGA) may detect these haemostatic factors. So TGA may be an additional tool for classifying the actual severity of haemophilia. Our group is carrying out correlation tests between FVIII and TGA in platelet-poor and -rich plasmas (PPP and PRP, respectively). PRP has the inconvenience that must be done freshly soon after blood extraction. Our aim is to study the differences between TGA performed with fresh and frozen PRP and PPP and its implementation in multicenter studies. We included 70 patients with severe haemophilia A in prophylactic treatment. Venous blood drawing was obtained prior to administration of FVIII, at the trough levels. FVIII measurement and TGA were performed in fresh and frozen PRP and PPP. The platelet absence caused a significant decrease in TGA although PPP and PRP correlated well. Frozen samples gave different results in PPP, but there were no significant differences between fresh and frozen PRP. This fact enables using frozen PRP in multicenter studies with a TGA-specialized laboratory for reclassifying haemophilia severity and for pharmacokinetic studies with TGA.

Quality assessment of genetic markers used for therapy stratification.

PURPOSE: Therapy stratification based on genetic markers is becoming increasingly important, which makes commitment to the highest possible reliability of the involved markers mandatory. In neuroblastic tumors, amplification of the MYCN gene is an unequivocal marker that indicates aggressive tumor behavior and is consequently used for therapy stratification. To guarantee reliable and standardized quality of genetic features, a quality-assessment study was initiated by the European Neuroblastoma Quality Assessment (ENQUA; connected to International Society of Pediatric Oncology) Group. MATERIALS AND METHODS: One hundred thirty-seven coded specimens from 17 tumors were analyzed in 11 European national/regional reference laboratories using molecular techniques, in situ hybridization, and flow and image cytometry. Tumor samples with divergent results were re-evaluated. RESULTS: Three hundred fifty-two investigations were performed, which resulted in 23 divergent findings, 17 of which were judged as errors after re-evaluation. MYCN analyses determined by Southern blot and in situ hybridization led to 3.7% and 4% of errors, respectively. Tumor cell content was not indicated in 32% of the samples, and 11% of seemingly correct MYCN results were based on the investigation of normal cells (eg, Schwann cells). Thirty-eight investigations were considered nonassessable. CONCLUSION: This study demonstrated the importance of revealing the difficulties and limitations for each technique and problems in interpreting results, which are crucial for therapeutic decisions. Moreover, it led to the formulation of guidelines that are applicable to all kinds of tumors and that contain the standardization of techniques, including the exact determination of the tumor cell content. Finally, the group has developed a common terminology for molecular-genetic results.

Clinical usefulness of KRAS mutational analysis in the diagnosis of pancreatic adenocarcinoma by means of endosonography-guided fine-needle aspiration biopsy.

AIM: To establish the usefulness of KRAS mutational analysis in the diagnosis of pancreatic adenocarcinoma by comparing this technique with conventional cytology in aspirates obtained by endosonography-guided fine-needle aspiration. METHODS: All consecutive patients with pancreatic focal lesions undergoing endosonography-guided fine-needle aspiration were included. Samples were obtained with the concurrence of an attendant cytopathologist. Detection of codon-12 KRAS mutations was performed by the restriction fragment length polymorphism-polymerase chain reaction method. The effectiveness of conventional cytology, KRAS mutational analysis and their combination was established with respect to the definitive diagnosis. A cost-effectiveness analysis was also performed. RESULTS: Thirty-three patients had pancreatic adenocarcinoma and 24 patients had other lesions. A total of 136 samples was obtained. In patients in whom specimens were adequate (93% for cytology; 100% for mutational analysis), the specificity of both techniques was 100%, whereas the sensitivity favoured cytology (97% vs. 73%). When inadequate samples were considered as misdiagnosed, a combination of both techniques reached the highest overall accuracy (cytology, 91%; mutational analysis, 84%; combination of both, 98%). CONCLUSIONS: Cytology from aspirates obtained by endosonography-guided fine-needle aspiration is the most precise single technique for the diagnosis of pancreatic adenocarcinoma. However, when adequate specimens are not available to reach a cytological diagnosis, the addition of KRAS mutational analysis represents the best strategy.

[Secondary costs due to nosocomial infections in 2 pediatric intensive care units]. / Costos secundarios por infecciones nosocomiales en dos unidades pediátricas de cuidados intensivos.

OBJECTIVE: We estimated associated costs to nosocomial infections in two pediatric intensive care units in Mexico City. MATERIAL AND METHODS: A transversal study in the neonatal (NICU) and pediatric (PICU) intensive care units, was done. We reviewed use and cost of diagnostic procedures, medications, and excess of hospital stay. RESULTS: There were 102 infections, 46 in the NICU and 56 in the PICU. The average cost per infection was $11,682 USD and the overall expense was 1,184.71 USD. Infected children had an excess of hospital stay of 9.6 days, 13.7 more laboratory tests and 3.3 more cultures. Hospital stay represented 97% of the overall cost. CONCLUSIONS: This is one of the first estimations of nosocomial infections cost done in Mexico. These results justify the introduction of infection control programs to decrease these complications.

[Nosocomial infections and quality of health care]. / Las infecciones nosocomiales y la calidad de la atención médica.

The main objective of a hospital-acquired infections control program is to decrease the risk of acquisition and the morbidity and costs associated. The organization of a team with technical and humanistic leadership is essential. Every infection control program must also develop strategies that allow: a) identification of the problems, b) to establish the importance of each one, c) to determine their causes, d) to develop solutions and e) the evaluation of the recommended solutions. The development of technical and humanistic abilities by the leader and the members of the team, and the use of the tools mentioned above have produced the only validate and highly effective program of quality improvement in the hospital.

[Surveillance and the costs related to the prescription of antimicrobials in a pediatric hospital]. / Vigilancia y costos relacionados con la prescripción de antimicrobianos en un hospital pediátrico.

INTRODUCTION: Important considerations in the selection of an antimicrobial agent include the indications for its prescription, its efficacy, toxicity and cost. However, is has been observed that up to 50% of these prescriptions are inappropriate. OBJECTIVE: To evaluate the utilization patterns and costs of restricted antimicrobial agents requiring authorization of the infectious diseases service for their prescription, in a tertiary care pediatric hospital through the use of a computer software antimicrobial use surveillance program designed by the investigators. MATERIAL AND METHODS: A one-year longitudinal descriptive and prolective survey was conducted in all patients requiring antimicrobial therapy during their hospitalization. Information on the prescription and costs of antimicrobial agents was recorded from both the medical chart and pharmacy registers. RESULTS: A total of 748 patients were prescribed at least one restricted antimicrobial agent, corresponding to 9.6% of the total number of patients hospitalized and 63.4% of the antibiotic expenditures during the study. CONCLUSIONS: Hospital surveillance of antimicrobial prescription is useful for detecting antimicrobial prescription patterns and costs. These types of studies are inexpensive and of easy access when a computerized register is available in the pharmacy.

[Medical facilities for women and children infected with HIV]. / Infraestructura de atención y servicio para mujeres y niños infectados por el virus de la inmunodeficiencia humana.

The phenomena HIV infection and acquired immunodeficiency syndrome (AIDS) among women and children is an important concern for underdeveloped and developing countries and has became a Public Health problem. The medical facilities required for care of these patients are special and demand a comprehensive approach and multidisciplinary team-work. In this way, the patients would receive adequate care for their needs. This review includes the antecedents of the HIV infection among women and children. Mechanisms of transmission, and the current model of medical care that exists in some developed countries. Finally, a commentary is made in regards to the importance of continuing medical education of the health-term in order to confront the AIDS epidemic.