Life Cycle Thinking for the environmental and financial assessment of rice management systems in the Senegal River Valley.

Rice is a staple food in Senegal, which however imports more than 70% of the rice consumed annually to meet its domestic demand. Despite governmental efforts to increase rice self-sufficiency, both rice supply and yields remain low. Senegalese farmers face challenges related to irrigation infrastructure and fertiliser access, besides those derived from climate change. This study applies Life Cycle Assessment (LCA) combined with financial Life Cycle Costing (LCC) to evaluate alternative scenarios for rice management in the Senegal River Valley and identify sustainability hotspots and potential improvements. Specifically, rice cultivation in Ross Béthio (Saint Louis, Senegal) is assessed based on the observed agricultural practices during the dry seasons of 2016 and 2017. Two scenarios capturing conventional (CONV) and intensive (INT) practices are compared to two reference scenarios (SAED scenarios) according to the recommendations of the official agricultural advisory service. The INT scenario generates the lowest impacts per kg of paddy rice in seven out of thirteen impact categories, including climate change, freshwater and marine eutrophication, ozone depletion and water scarcity. This is due to the higher yields (7.4 t ha-1) relative to CONV (4.8 t ha-1) and the two reference SAED scenarios (6.0 t ha-1). The two latter scenarios show the lowest values in the remaining categories, although they also generate slightly lower profits than INT (138 € t-1 vs. 149 € t-1) due to increased labour costs for additional fertilisation treatments. The results from both LCA and LCC underline the importance of increasing yields to decrease environmental impacts and production costs of rice when estimated per kg of product. Well-designed fertiliser application doses and timing and increased mechanisation can deliver further environmental benefits. Additional improvements (e.g. in irrigation, crop rotations, straw management) could be considered to promote the long-term sustainability and profitability of rice production in Senegal. LCA in combination with financial LCC is identified as a decision-support tool for evaluating the sustainability of alternative crop management practices. Life Cycle Thinking can still benefit from experiential learning based on information exchange between farmers, researchers and extension agents to contribute to a sustainable agriculture and ultimately to food security in Africa.

'Ethnobiological equivocation' and other misunderstandings in the interpretation of natures.

In this contribution we seek to enrich the theoretical and methodological approaches of ethnobiology. The essay takes elements of Amerindian anthropology, classical ethnobiological studies and the freedoms provided by feminist philosophers to open up reflection. The central background of the essay is the method of "controlled equivocation" proposed by Viveiros de Castro (2004). We present a series of five ethnobiological equivocations ranging from the categorical equivocal, going through the subtle equivocal to the strictly ontological ones. The cases occurred in different territories of Argentina, including a case in an academic context. Through the fieldwork cases, we give an account of the origin of equivocations, the context for their emergence, which are the disciplinary nuances that cause them and even some academics' preconceptions. To inhabit the equivocation allows opening the possibilities of coexistence among people -and their respective worlds-, especially if these people are in different power positions. We propose the method of controlled equivocation as a theoretical-discursive tool, which permits us to rethink the current concepts of ethnobiology. Thus, we want to broaden the current definition of ethnobiology understood as a dialogue from different scientific points of view.

Nimotuzumab and radiotherapy for treatment of newly diagnosed diffuse intrinsic pontine glioma (DIPG): a phase III clinical study.

BACKGROUND: Diffuse intrinsic pontine glioma (DIPG) is a devastating cancer of childhood and adolescence. METHODS: The study included patients between 3 and 20 years with clinically and radiologically confirmed DIPG. Primary endpoint was 6-month progression-free survival (PFS) following administration of nimotuzumab in combination with external beam radiotherapy (RT). Nimotuzumab was administered intravenously at 150 mg/m2 weekly for 12 weeks. Radiotherapy at total dose of 54 Gy was delivered between week 3 and week 9. Response was evaluated based on clinical features and MRI findings according to RECIST criteria at week 12. Thereafter, patients continued to receive nimotuzumab every alternate week until disease progression/unmanageable toxicity. Adverse events (AE) were evaluated according to Common Terminology Criteria for Adverse Events (CTC-AE) Version 3.0 (CTC-AE3). RESULTS: All 42 patients received at least one dose of nimotuzumab in outpatient settings. Two patients had partial response (4.8%), 27 had stable disease (64.3%), 10 had progressive disease (23.8%) and 3 patients (7.1%) could not be evaluated. The objective response rate (ORR) was 4.8%. Median PFS was 5.8 months and median overall survival (OS) was 9.4 months. Most common drug-related AEs were alopecia (14.3%), vomiting, headache and radiation skin injury (7.1% each). Therapy-related serious adverse events (SAEs) were intra-tumoral bleeding and acute respiratory failure, which were difficult to distinguish from effects of tumor progression. CONCLUSIONS: Concomitant treatment with RT and nimotuzumab was feasible in an outpatient setting. The PFS and OS were comparable to results achieved with RT and intensive chemotherapy in hospitalized setting.

Profiling of volatile organic compounds produced by clinical Aspergillus isolates using gas chromatography-mass spectrometry.

Volatile organic compounds (VOCs) in exhaled breath may identify the presence of invasive pulmonary aspergillosis. We aimed to detect VOC profiles emitted by in vitro cultured, clinical Aspergillus isolates using gas chromatography-mass spectrometry (GC-MS). Three clinical Aspergillus isolates and a reference strain were cultured while conidiation was prevented. Headspace samples were analyzed using a standardized method. Breath samples of patients from which the cultures were obtained were checked for the presence of the VOCs found in vitro. Each Aspergillus isolate produced a distinct VOC profile. These profiles could not be confirmed in exhaled breath in vivo.

Properties of antibacterial nano textile for use in hospital environments / Propiedades del nanotextil antibacteriano para su uso en entornosm hospitalarios / Propriedades do nano têxtil antibacteriano para seu uso em meios hospitalares

Abstract In hospital environments, there are several problems related to Healthcare-Associated Infections (HAIs), contaminated hospital textiles, can contribute to the spread and transmission of (HAIs), due to retention of viruses and bacteria. The antibacterial metallic nanoparticles immersed in hospital textiles can allow reduction of microorganisms. This paper presents a technological surveillance of the principal properties of antibacterial nanotextiles to be used in hospital environments, based on international standards. Initially, the search equation was determined for "antibacterial" AND "nanoparticle." Subsequently, the main properties were selected, by means of a multiple authors' review. Afterwards, the properties were related to international standards. Finally, we present the results found associated to the materials used to develop nonwoven textiles, and their properties for hospital environments, the sizes of samples and also the equipment required for characterization.

Resumen En los ambientes hospitalarios, existen varios problemas relacionados con las infecciones asociadas a la atención de la salud (HAI, por sus siglas en inglés), los tejidos hospitalarios contaminados, pueden contribuir a la propagación y transmisión de los HAIs, debido a la retención de virus y bacterias. Las nanopartículas metálicas antibacterianas sumergidas en tejidos hospitalarios permiten reducir los microorganismos. Este documento presenta una vigilancia tecnológica de las principales propiedades del nanotextil antibacteriano para uso en ambientes hospitalarios, basados en estándares internacionales. Inicialmente, la ecuación de búsqueda se determinó "antibacteriano" Y "nanopartícula". Posteriormente, se seleccionaron las principales propiedades, mediante la revisión de diferentes autores. Luego, las propiedades se relacionaron con los estándares internacionales. Finalmente, se presentan los resultados encontrados asociados a los materiales utilizados para el desarrollo de materiales no tejidos y sus propiedades para ambientes hospitalarios, tamaños de muestras y también el equipo necesario para la caracterización.

Resumo Nos ambientes hospitaleiros, existem vários problemas relacionados com as infecções associadas à atenção da saúde (HAI), os tecidos hospitalários contaminados, podem contribuir à propagação e transmissão dos HAIs, devido à retenção de vírus e bactérias. As nano partículas metálicas antibacterianas submergidas em tecidos hospitalários permitem reduzir os microorganismos. Este documento apresenta uma vigilância tecnológica das principais propriedades do nano têxtil antibacteriano para uso em ambientes hospitalários, baseados em padrões internacionais. Inicialmente, a equação de busca determinou-se "antibacteriano" e "nano partícula". Posteriormente, selecionaram-se as principais propriedades, mediante a revisão de diferentes autores. Posteriormente, as propriedades relacionaram-se com os padrões internacionais. Finalmente, apresentam-se os resultados encontrados associados aos materiais utilizados para o desenvolvimento de tecidos não tecidos e suas propriedades para ambientes hospitaleiros, tamanhos de amostras e também a equipe necessária para a caracterização.

Cognition, culture and utility: plant classification by Paraguayan immigrant farmers in Misiones, Argentina.

BACKGROUND: This study was conducted in three rural communities of small farmers of Paraguayan origin living in the province of Misiones, Argentina. These Criollos (Mestizos) hail chiefly from departments located in the east of Paraguay, where the climate and flora have similar characteristics as those in Misiones. These ecological features contribute to the continuation and maintenance of knowledge and practices related to the use of plants. METHODS: Fieldwork was conducted between September 2014 and August 2015. Forty five informants from three rural localities situated along the Parana River participated in an ethno-classification task. For the classification event, photographs of 30 medicinal and edible plants were chosen, specifically those yielding the highest frequency of mention among the members of that community (based on data obtained in the first stage of research in 2014). Variation in local plant classifications was examined and compared using principal component analysis and cluster analysis. RESULTS: We found that people classify plants according to application or use (primarily medicinal, to a lesser extent as edible). Morphology is rarely taken into account, even for very similar and closely-related species such as varieties of palms. In light of our findings, we highlight a dominant functionality model at work in the process of plant cognition and classification among farmers of Paraguayan origin. Salient cultural beliefs and practices associated with rural Paraguayan plant-based medicine are described. Additionally, the manner by which residents' concepts of plants articulate with local folk epistemology is discussed. CONCLUSIONS: Culturally constructed use patterns ultimately override morphological variables in rural Paraguayans' ethnobotanical classification.

Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation.

Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

Using Positive Deviance in the prevention and control of MRSA infections in a Colombian hospital: a time-series analysis.

Positive Deviance (PD) is a process to achieve a social and cultural change. This strategy has been used for the control of methicillin-resistant Staphylococcus aureus (MRSA) infection in some health institutions in the United States, but has rarely been adopted in institutions from developing countries where resources are limited. We describe our experience of PD in the control of healthcare-associated infections (HAIs) due to MRSA in a Colombian hospital with the aim of reducing HAI rates through a cultural change in processes. A time-series study was conducted based on the MRSA-HAI rate and the number of months with zero MRSA infections before and after application of PD (2001-2012). On comparing the pre-intervention and intervention periods, the mean overall rates of MRSA-HAI was 0·62 and 0·36, respectively (P = 0·0005); the number of months with zero MRSA-HAIs were 3/70 and 12/74 (odds ratio 0·264, 95% confidence interval 0·078-0·897); the percentage of MRSA-HAIs was 53·2% and 41·0%. These results are consistent with other published data. Implementation of PD was associated with a significant reduction of MRSA-HAIs, it did not involve high costs and the changes have been lasting.

Búsqueda de kpi de facility management para administrar la infraestructura hospitalaria en colombia / Search for facility management's kpis to manage the hospitals infrastructure in colombia / Busca dos indicadores chave de desempenho, kpis, da gestão de instalações para gerenciar a infra-estrutura dos hospitais na colômbia

En la actualidad, la gestión de activos físicos hospitalarios está adquiriendo importancia debido al tamaño de sus edificaciones, la relevancia social, los costos operativos y de mantenimiento. El propósito de este documento es identificar los Indicadores Clave de Desempeño (KPI por sus siglas en inglés) y clasificarlos en aspectos específicos de medición de desempeño de la infraestructura. La carencia de una clasificación adecuada de los indicadores de desempeño impide el uso general de las medidas de desempeño para el Facility Management (FM por sus siglas en inglés) en el sector de la salud. En este artículo se muestra una breve reseña del estado del arte de los KPIs del FM, basada en una búsqueda bibliográfica teniendo en cuenta autores internacionales y analizando sus enfoques y categorizaciones. Los autores de este trabajo presentan un resumen de la categorización y del enfoque de la gestión de la infraestructura hospitalaria basándose en los documentos examinados, lo que puede ofrecer un uso más práctico para los profesionales del FM en el sector de la salud. Los Facility Managers deben abordar demasiada información lo que dificulta llevar a cabo un análisis más profundo de los indicadores clave de desempeño en investigaciones futuras e intentar generar indicadores que coincidan con los objetivos de la organización y que conduzcan a una gestión de la infraestructura de la salud más inteligente y eficiente.

Nowadays, hospital physical asset management is gaining importance due to the size of its buildings, social relevance, operational and maintenance costs. The purpose of this document is to identify the Key Performance Indicators (KPIs) and categorize them into specific aspects of measurement of infrastructure performance. The lack of an adequate classification of performance indicators prevents the widespread use of performance measures for Facility Management (FM) in the health sector. This article shows a brief review of the state of art of FM KPIs based on a bibliographic search, taking into account international authors and reviewing their approaches and categorizations. The authors of this paper present a summary of the categorization and approach of hospital infrastructure management based on the documents examined, which may offer a more practical use for FM professionals in the healthcare sector. Facility Managers have to tackle too much data, which hinders to carry out a deeper analysis of the key performance indicators in future researches, attempting to generate indicators that match with the organization objectives and that lead to a more intelligent and efficient health infrastructure management.

Hoje em dia, a gestão de ativos físicos hospitalários está a ganhar importância devido ao tamanho de seus prédios, sua relevância social, os custos operacionais e de manutenção. O propósito deste documento é identificar os Indicadores Chave de Desempenho (KPIs, em inglês) e categorizá-los em aspectos específicos da medição do desempenho da infra-estrutura. A falta de uma adequada classificação dos indicadores de desempenho impede o uso generalizado de medidas de desempenho para a Gestão de Instalações (FM, em inglês) no sector da saúde. Este artigo apresenta uma breve revisão do estado da arte dos KPI de FM baseados numa procura bibliográfica, tendo em conta aos autores internacionais e revisando seus enfoques e categorizações. Os autores deste trabalho apresentam um resumo da categorização e enfoque da gestão da infra-estrutura hospitalaria a partir dos documentos examinados, o que pode oferecer um uso mais prático para os profissionais da Gestão de Instalações - FM no sector da saúde. Os gestores de instalações têm que abordar bastante dados, o que dificulta uma análise mais profunda dos indicadores chave de rendimento em futuras investigações, tentando gerar indicadores que coincidam com os objetivos da organização e que conduzam a uma gestão da infra-estrutura de saúde mais inteligente e eficiente.

Evaluation of Bone Mineral Density in Patients with Type 1 Gaucher Disease in Argentina.

The purpose of this study was to evaluate the frequency of osteoporosis (OP) in patients with Gaucher disease (GD) in Argentina. GD patients from 28 centers were consecutively included from April 2012 to 2014. Bone mineral density (BMD) was determined by dual X-ray absorptiometry in the lumbar spine and the femoral neck or the total proximal femur for patients ≥20 yr of age, and by whole-body scan in the lumbar spine in patients <20 yr of age. In children, mineral density was calculated using the chronological age and Z height. OP diagnosis was determined following adult and pediatric official position of the International Society for Clinical Densitometry. A total of 116 patients were included, of which 62 (53.5%) were women. The median age was 25.8 yr. All patients received enzyme replacement therapy, with a median time of 9.4 yr. Normal BMD was found in 89 patients (76.7%), whereas low bone mass (LBM) or osteopenia was found in 15 patients (13%) and OP in 12 patients (10.3%). The analysis of the pediatric population revealed that 4 patients (9.3%) had LBM and 3 (7%) had OP (Z-score ≤ -2 + fractures height-adjusted by Z), whereas in the adult population (n = 73), 11 patients (15%) had LBM or osteopenia and 9 (12.3%) had OP. Bone marrow infiltration and the presence of fractures were significantly correlated with the presence of OP (p = 0.04 and <0.001, respectively). This is the first study in Argentina and in the region describing the frequency of OP or LBM in GD patients treated with imiglucerase using the official position of the International Society for Clinical Densitometry.

Molecular and morphometric characterization of two dental morphs of Saccodon dariensis (Parodontidae).

This study reports for the first time the roles of genetic and body phenotypic variation in two Saccodon dariensis dental morphs. Results showed a lack of ancient mitochondrial differentiation between morphs and body variations concordant with genetic polymorphism or differential plastic responses to diet quality and foraging strategies of S. dariensis.

Variations of body geometry in Brycon henni (Teleostei: Characiformes, Bryconidae) in different rivers and streams.

Landmark-based geometric morphometrics were used to evaluate the effect of flow and depth in the water column on the body size and shape of Brycon henni from river and stream populations. The dendrogram showed that stream populations clustered apart and showed significantly smaller centroid size and narrower bodies than river populations, indicating a role for flow and depth on whole body morphometric variation. The results are concordant with morphometric variation described in other fish species and provide the first insights into phenotypic variation in natural populations of B. henni.

A Mayan founder mutation is a common cause of deafness in Guatemala.

Over 5% of the world's population has varying degrees of hearing loss. Mutations in GJB2 are the most common cause of autosomal recessive non-syndromic hearing loss (ARNHL) in many populations. The frequency and type of mutations are influenced by ethnicity. Guatemala is a multi-ethnic country with four major populations: Maya, Ladino, Xinca, and Garifuna. To determine the mutation profile of GJB2 in a ARNHL population from Guatemala, we sequenced both exons of GJB2 in 133 unrelated families. A total of six pathogenic variants were detected. The most frequent pathogenic variant is c.131G>A (p.Trp44*) detected in 21 of 266 alleles. We show that c.131G>A is associated with a conserved haplotype in Guatemala suggesting a single founder. The majority of Mayan population lives in the west region of the country from where all c.131G>A carriers originated. Further analysis of genome-wide variation of individuals carrying the c.131G>A mutation compared with those of Native American, European, and African populations shows a close match with the Mayan population.

ADAPTACIÓN DE UNA METODOLOGÍA PARA LOS PROCESOS DE ADQUISICIÓN DE SISTEMAS DE AIRE ACONDICIONADO EN SERVICIOS DE SALUD / ADAPTATION OF A METHODOLOGY FOR PROCUREMENT PROCESSES OF AIR CONDITIONING SYSTEMS IN HEALTH SERVICES / ADAPTAÇÃO DE UMA METODOLOGIA PARA OS PROCESSOS DE AQUISIÇÃO DE SISTEMAS DE AR CONDICIONADO NOS SERVIÇOS DE SAÚDE

La adquisición efectiva de tecnologías en salud está enfocada al mejoramiento de la calidad y eficiencia, en la prestación de los servicios de salud ya que es un proceso complejo. Dentro de este proceso se destacan la evaluación técnica y clínica. El objetivo de la evaluación de tecnologías en salud es recolectar, analizar y sintetizar información y conocimiento para contribuir a mejorar la toma de decisiones en la práctica médica y en las políticas de salud. En este trabajo se presenta una evaluación en procesos de adquisición de sistemas de aire acondicionado en servicios de salud, basada en la definición de variables en campos de evaluación; dichas variables cuantifican lo técnico y funcional del equipo en su entorno clínico y económico. De igual manera, se pretende impulsar en los departamentos de ingeniería clínica la implementación de metodologías de amplia aceptación y efectividad que tienen como objetivo primordial la adquisición de tecnologías en salud basados en la evidencia.

The effective acquisition of health technologies is focused on improving the quality and efficiency in the provision of health services as it is a complex process. Within this process the technical and clinical evaluation are highlighted. The objective of the health technology assessment is to collect, analyze, and synthesize information and knowledge to help improve decisionmaking in medical practice and health policy. This paper presents an evaluation process of acquiring air conditioning systems in health care facilities, based on the definition of variables into a field evaluation; these variables quantify the technical and functional team in their clinical and economic environment. Similarly, it is intended to promote in clinical engineering departments, the implementation of widely accepted methodologies and effectiveness that have, as its primary objective, the acquisition of technologies in evidence-based health.

A aquisição efetiva das tecnologias da saúde é focada na melhoria da qualidade e eficiência na prestação de serviços de saúde, pois é um processo complexo. Dentro deste processo se destaca a avaliação técnica e clínica. O objetivo da avaliação de tecnologias em saúde é coletar, analisar e sintetizar informações e conhecimentos para ajudar a melhorar a tomada de decisão na prática médica e a política de saúde. Neste trabalho se apresenta um processo de avaliação de aquisição de sistemas de ar condicionado nos serviços de saúde, com base na definição de variáveis no campo da avaliação; essas variáveis vão quantificar o técnico e o funcional do equipamento em seu ambiente clínico e econômico. Da mesma forma, pretende-se promover nos departamentos clínicos de engenharia a aplicação de metodologias amplamente aceitas e eficácia que têm como objetivo principal a aquisição de tecnologias em saúde baseada em evidências.

MODELO INTEGRADO DE GESTIÓN DE ACTIVOS HOSPITALARIOS BASADO EN LA PAS 55 / INTEGRATED MANAGEMENT MODEL OF HOSPITAL ASSETS BASED ON PAS 55 / MODELO INTEGRADO DE GESTÃO DE ATIVOS HOSPITALARES COM BASE NA PAS 55

Este trabajo presenta un modelo de gestión de activos hospitalarios basado en la PAS 55, que se desarrolló a partir de la homologación de los requerimientos legales, de acreditación nacional e internacional y las recomendaciones de la Organización Mundial de la Salud OMS, la Pan American Health Organization PAHO y el Ministerio de la Protección Social. Se busca con este proyecto involucrar el ciclo de vida de los activos hospitalarios, plantear herramientas para la toma de decisiones frente a la adquisición y reposición de activos hospitalarios, plantear de la mano con los criterios del sector salud metodologías de evaluación de criticidad de los activos físicos hospitalarios, y definir metodologías y herramientas que faciliten la adecuada gestión de las instituciones frente al cuidado de los activos físicos y los riesgos asociados a los mismos.

This paper presents a management model on hospital assets based on PAS 55, which evolved from the approval of the legal requirements, national and international accreditation, and the recommendations of the World Health Organization -WHO, the PAHO, Pan American Health Organization and the Ministry of Social Protection of Colombia. This project seeks to include lifecycle of hospital assets, introduce new tools for decision making to the acquisition and replacement of hospital assets, uniting the criteria of health methodologies sector in evaluation of critical hospital critical assets and defining methodologies and tools to facilitate the adequate management of institutions for physical assets care and associated risks.

Neste artigo se apresenta um modelo de gestão de ativos hospitalares baseado na PAS 55, que evoluiu a partir da aprovação dos requisitos legais, de acreditação nacional e internacional e as recomendações da Organização Mundial da Saúde OMS, a Pan American Health Organization PAHO e do Ministério da Proteção Social. Destina-se com este projeto involucrar o ciclo de vida de ativos hospitalários, planejar ferramentas para fazendo para a toma de decisões frente para a aquisição e substituição de ativos hospitalários, planejar da mão para os critérios de avaliação do sector saúde metodologias de avaliação nos ativos físicos hospitalários criticidade e definir metodologias e ferramentas que facilitem o adequada gestão das instituições frente o cuidado de ativos físicos e risos associados a eles.

Expression of helminth genes in Leishmania: an experimental transfection system to test immunological function.

Functional analysis of genes from parasitic helminths requires, at the present time, heterologous expression. We have adapted the well-characterized system of transfection in Leishmania protozoal parasites, as a means of analysing the effect of single filarial genes on the mammalian immune system. For example, testing the function of the Brugia malayi abundant larval transcript (ALT) gene-transfected Leishmania mexicana were found to be significantly more virulent in macrophages in vitro. The course of infection in vivo is also aggravated by expression of the ALT gene. Examples are also given of transgenes which reduced in vitro growth within macrophages, as well as others which exert no effect on the protozoal parasitism. Thus, Leishmania transfection provides a tractable system to analyse helminth gene function within the context of the host immune system.

Construction of an immunotoxin with the pore forming protein StI and ior C5, a monoclonal antibody against a colon cancer cell line.

Sticholysin I (StI), a potent cytolysin isolated from the sea anemone Stichodactyla helianthus, was linked to the monoclonal antibody (mAb) ior C5. StI acts by forming hydrophilic pores in the membrane of the attacked cells leading to osmotic lysis. ior C5 is a murine IgG1, which recognizes the tumor associated antigen (TAA) ior C2. The cytolysin and the mAb were coupled by using the heterobifunctional cross-linking reagent sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC). Two hybrid molecules composed by one ior C5 and one or two StI molecules were obtained (named conjugated I and II, respectively). The purified conjugates were evaluated by a binding affinity assay against an ior C2-positive colon cancer cell line (SW948). Both molecules were able to recognize the antigen (Ag) in the same way that unconjugated ior C5 does. The activity of both conjugates against human erythrocytes and SW948 cells was assessed. They lost most of their hemolytic activity but their residual activity was very similar. Nevertheless, when their cytotoxicity was studied on the SW948 cell line, only conjugate II killed efficiently the cells, indicating a specific mAb-Ag interaction. In this chimeric molecule the ratio between the cytotoxic and the hemolytic activity was larger than that of the free cytolysin. This fact indicates an increase of the specificity of the toxic effect toward the SW948 cell line and consequently an increase of the difference between its hemolytic and cytotoxic doses. The results herein support the feasibility of directing StI to the surface of cancer cells expressing ior C2 Ag via the mAb ior C5.

Factors affecting pharmacokinetics of monoclonal antibodies: A review article.

Factors affecting the successful therapy of malignant diseases include the antibody dose used and the schedule of administration, the half-life and fast blood clearance of the antibodies, the presence of circulating antigen, poor tumor penetration of the high/molecular-weight monoclonal antibody (mAb) and the way in which these molecules are catabolized. To circumvent these limitations and achieve higher uptake, increased tumor penetration, faster blood clearance and longer retention in the tumors, there is a need to generate mAbs suitable for diagnosis as well as therapy and to develop novel strategies to increase the efficacy of immunotherapeutic treatments. There is a lack of knowledge about many aspects of the physiological function and metabolism of antibodies. This paper is intended to discuss factors that affect the pharmacokinetics of mAbs in human subjects with the purpose of forming possible strategies to optimize this approach for tumor diagnosis and therapy.

(99m)Tc-labeled murine ior C5 monoclonal antibody in colorectal carcinoma patients: pharmacokinetics, biodistribution, absorbed radiation doses to normal organs and tissues and tumor localization.

Monoclonal Antibody (mAb) ior C5 is a murine IgG(1) that recognizes the tumor associated antigen (TAA) ior C2, a cell surface O-linked glycoprotein carbohydrate chain not present in most normal tissues and homogeneously expressed in the cytoplasm of normal colon epithelium and heterogeneously expressed in more than 83% of primary colorectal carcinomas. This study was designed to investigate the pharmacokinetics, biodistribution and the absorbed radiation doses of (99m)Tc-labeled mAb ior C5 antibody in colorectal tumor patients. Ten patients were administered 3 mg of anti-O-linked glycoprotein carbohydrate chain TAA ior C2 murine monoclonal antibody ior C5 radiolabeled with (99m)Tc activity of 1435.0 +/- 123 MBq by intravenous (i.v.) bolus infusion. Blood and urine samples were collected from 4 out of 10 patients at timed intervals from 10 min and up to 24 h after injection of the (99m)Tc-labeled mAb ior C5 for pharmacokinetic studies. Whole body images were taken in 5 out of 10 patients for quantitative normal organ biodistribution and dosimetry studies and planar anterior and posterior and SPECT images were taken in 5 out of 10 patients for tumor localization. Mean absorbed doses were estimated using the methods developed by the Medical Internal Radiation Dose (MIRD) committee. The effective dose equivalent (EDE) and effective dose (ED) were calculated as prescribed in International Commission on Radiological Protection (ICRP) publications 30 and 60. Plasma disappearance curves of (99m)Tc-labeled murine antibody ior C5 were best fit by a two-compartment model in all patients with (t(1/2alpha)) of 4.32 +/- 2.18 h and (t(1/2beta) of 32.6 +/- 3.82 h. Among the main target organs, accumulation of the radiolabeled antibody was found in liver (9.38 +/- 0.80%), heart (8.92 +/- 0.94%) and spleen (1.37 +/- 0.30%) at 5 min post-administration. These values were reduced at 24 h to (5.91 +/- 0.73%) and (0.62 +/- 0.22%), respectively, for the heart and spleen and increased to (9.78 +/- 1.99%) for liver. Estimates of radiation absorbed dose to normal organs in rad/mCi administered were: whole body, 0.0181 +/- 0.0017; heart wall, 0.0768 +/- 0.0090; kidneys, 0.0530 +/- 0.0260; liver, 0.0565 +/- 0.0109 and spleen, 0.0540 +/- 0.0128. The effective dose equivalent and effective dose estimates for adults were 0.0314 +/- 0.0031 and 0.0249 +/- 0.0027 rem/mCi administered. This feasibility study indicates that the O-linked glycoprotein carbohydrate chain TAA ior C2 is expressed in primary and metastatic colorectal carcinomas and shows very limited expression in normal adult tissues. The very good pattern of biodistribution of (99m)Tc-labeled mAb ior C5 in patients will allow imaging of colorectal carcinoma lesions.

188Re-labeled anti-epidermal growth factor receptor humanized monoclonal antibody h-R3: labeling conditions, in vitro and in vivo stability.

The use of antibodies as targeting agents for the delivery of radioisotopes to tumors is an appealing concept that has received widespread attention since the advent of monoclonal antibody (mAb) technology. The present study describes the (188)Re-direct labeling of anti-epidermal growth factor receptor (EGF-R) humanized mAb h-R3; the analytical methods for quality control of radiopharmaceuticals such as instant thin layer chromatography-silica gel (ITLC-SG); the immunoreactivity and biological recognition of the target antigen assessment of the radiolabeled molecule using flow cytometry analysis; in vitro stability studies using saline 0.9% solution, cysteine, diethylenetriaminepentaacetic acid (DTPA), human serum and human serum albumin (HSA) 1% challenge; and the assessment of in vivo stability through biodistribution studies in normal Balb/c mice. No fragmentation of the reduced molecules was found using 2-ME as a reducing agent. Labeling efficiency was greater than 98.5 +/- 0.6% of rhenium-188 (188Re) bound to IgG1 after 5 h, as determined by paper chromatography in saline 0.9% solution. Radiocolloids determined by albumin impregnated ITLC was 1.04 +/- 0.07% in all cases. The biological activity measured by flow cytometry analysis showed an immunoreactivity fraction and the biological recognition of the target antigen overexpressed on H-125 human lung adenocarcinoma cell line greater than 87%. Challenge studies with cysteine, DTPA, human serum and HSA 1% demonstrated no evidence of transcomplexation of 188Re to DTPA or HSA and showed that 30% and 85% of the 188Re-radiolabeled was transcomplexed to human serum and to 100 mM cysteine after 24 h for human serum and 1 h incubation for cysteine at 37 masculine C, respectively. Biodistribution studies indicated no accumulation of the radiolabeled antibodies in normal organs.