Método indirecto para la verificación del intervalo de referencia del perfil lipídico / Indirect method for the verification of the lipid profile reference range

Introducción: La dislipidemia es uno de los problemas más frecuentes en los niños y adolescentes y su estudio es importante debido a su fuerte correlación con la enfermedad cardiovascular aterosclerótica en adultos. Muchos países desarrollaron valores de referencia nacionales investigando los lípidos séricos utilizando datos basados en la población nacional propia. Nuestro objetivo fue verificar el intervalo de referencia del perfil lipídico calculando las curvas de percentiles a través del método indirecto en nuestra población pediátrica. Materiales y métodos: Se analizaron los resultados de nuestra base de datos utilizando el método indirecto. Luego de aplicar filtros y criterios de exclusión se calcularon los percentiles 25, 50, 75, 95 y 99 para colesterol total (CT), colesterol HDL (C-HDL), colesterol no HDL (C-no-HDL), triglicéridos (TG) y colesterol LDL (C-LDL) y para el C-HDL además se calculó el percentil 10. El valor de referencia para el cambio (RCV) se utilizó para determinar si existía diferencia clínicamente significativa entre los valores de percentiles obtenidos y los utilizados en el consenso de la SAP. Resultados: No se evidenció diferencia clínicamente significativa contra los valores propuesto por la SAP, excepto para los TG para las edades 1,5,7 años en el percentil 95 y para la edad de 8 años en el percentil 75 y 95; para el C-HDL en el percentil 10 para las edades 1,16 y 17 años. Discusión: Se obtuvieron los percentiles de los lípidos y se compararon con los valores de referencia utilizados por el consenso en el que están basados las guías (AU)

Introduction: Dyslipidemia is one of the most common problems in children and adolescents and its study is important because of its strong correlation with atherosclerotic cardiovascular disease in adulthood. Many countries have developed national reference values investigating serum lipids using data based on their own national population. Our aim was to verify the lipid profile reference range by calculating percentile curves through the indirect method in our pediatric population. Materials and methods: The results of our database were analyzed using the indirect method. After applying filters and exclusion criteria, the 25th, 50th, 75th, 95th, and 99th percentiles were calculated for total cholesterol (TC), HDL cholesterol (HDL-C), non-HDL cholesterol (non-HDL-C), triglycerides (TG), and LDL cholesterol (LDL-C); for HDL-C, the 10th percentile was also calculated. The reference change values (RCV) were used to determine whether there was a clinically significant difference between the percentile values obtained and those used in the consensus of the Argentine Association of Pediatrics (SAP). Results: There was no clinically significant difference with the values proposed by the SAP, except for TG for ages 1, 5, and 7 years at the 95th percentile and for age 8 years at the 75th and 95th percentile; and for HDL-C at the 10th percentile for ages 1, 16, and 17 years. Discussion: Lipid percentiles were obtained and compared with the reference values used by the consensus on which the guidelines are based (AU)

Análisis clínicos en pediatría: la necesidad de minimizar las horas de ayuno / Clinical analysis in pediatrics: the need to minimize fasting hours

Los informes de laboratorio tienen impacto en las decisiones médicas. El ayuno es un factor preanalítico "controlable" que influye en los distintos parámetros bioquímicos. El objetivo del presente trabajo es poner en discusión la realización en pediatría de análisis clínicos con la indicación de un ayuno fisiológico , analizando resultados obtenidos por diferentes autores y evaluando las diferencias clínicas encontradas según los criterios de calidad establecidos por el laboratorio de Química Clínica. La mayoría de los individuos durante el día se encuentran en estado postprandial. Los resultados del perfil lipídico en ayunas no representan las concentraciones reales promedios de los lípidos plasmáticos de un paciente. El ayuno no sería crítico en la etapa de pesquisa , pero puede ser relevante para establecer un diagnóstico certero o inicio de tratamiento. En el caso de la glucemia si se indica en el control rutinario del paciente, y no hay sospecha de alteraciones en el metabolismo de los hidratos de carbono la glucemia sin ayuno puede ser solicitada comparando la misma con valores de corte adecuado. Las diferentes guías nacionales e internacionales recomiendan que la elección de la métrica para la evaluación, control y seguimiento de pacientes con diagnóstico de diabetes se realicen según el objetivo terapéutico. En los trabajos analizados, observamos que varios parámetros bioquímicos presentaron diferencias estadísticas, aunque las diferencias clínicas no fueron relevantes y permanecieron dentro de los intervalos de referencia. El factor limitante para evaluar parámetros bioquímicos sin ayuno es la falta de valores de referencia adecuados. Hay evidencia suficiente para que tanto el perfil lipídico, la glucemia como el resto de los parámetros bioquímicos del laboratorio de química clínica, sean solicitados con la indicación de un ayuno fisiológico de 2, 4 o 6 horas, dependiendo siempre del motivo de consulta y/o la edad del paciente. Es esencial extender la evaluación a otros analitos en población pediátrica, así como evaluar nuevos puntos de corte para parámetros bioquímicos sin ayuno (AU)

Laboratory reports have an impact on medical decision-making. Fasting is a "controllable" preanalytical factor that influences the different biochemical parameters. The aim of this study is to discuss the performance of clinical analyses in pediatrics with the indication of physiological fasting, analyzing results obtained in different disciplines, and evaluating the clinical differences found according to the quality criteria established by the clinical chemistry laboratory. During the day, most patients are in a postprandial state. Fasting lipid profile results do not represent the actual average plasma lipid concentrations of a patient. Fasting would not be critical in the screening stage, but it may be relevant to establish an accurate diagnosis or initiate treatment. Regarding glycemia, if it is indicated in the routine control of the patient and there is no suspicion of alterations in carbohydrate metabolism, non-fasting glycemia can be requested, comparing it with adequate cut-off values. Different national and international guidelines recommend that the choice of metrics for the evaluation, control, and follow-up of patients with diabetes should be made according to the therapeutic objective. In the studies analyzed, we found that several biochemical parameters presented statistical differences, although the clinical differences were not relevant and remained within the reference range. The limiting factor in the evaluation of biochemical parameters without fasting is the lack of adequate reference values. There is sufficient evidence that the lipid profile, glycemia, and the remaining biochemical parameters of the clinical chemistry laboratory should be requested with the indication of a physiological fast of 2, 4, or 6 hours, always depending on the reason for consultation and/or the patient's age. It is essential to extend the evaluation to other analytes in the pediatric population, as well as to evaluate new cut-off points for biochemical parameters without fasting (AU)

Field efficacy of Metarhizium anisopliae oil formulations against Rhipicephalus microplus ticks using a cattle spray race.

Rhipicephalus microplus tick is the main ectoparasite of cattle in Brazil. The exhaustive use of chemical acaricides to control this tick has favored the selection of resistant tick populations. Entomopathogenic fungi, as Metarhizium anisopliae, has been described as a potential biocontroller of ticks. Therefore, the aim of this study was to evaluate the in vivo efficacy of two oil based formulations of M. anisopliae for the control of the cattle tick R. microplus under field conditions using a cattle spray race as a method of treatment. Initially, in vitro assays were carried out with an aqueous suspension of M. anisopliae, using mineral oil and/or silicon oil. A potential synergism between oils and fungus conidia for tick control was demonstrated. Additionally, the usefulness of silicon oil in order to reduce mineral oil concentration, while improving formulation efficacy was illustrated. Based on the in vitro results, two formulations were selected for use in the field trial: MaO1 (107 conidia/mL plus 5% mineral oil) and MaO2 (107 conidia/mL plus 2.5% mineral oil and 0.01% silicon oil). The adjuvants concentrations (mineral and silicon oils) were chosen since preliminary data indicate that higher concentrations caused significant mortality in adult ticks. For this, 30 naturally infested heifers were divided into three groups based on previous tick counts. The control group did not receive treatment. The selected formulations were applied on animals using a cattle spray race. Subsequently, tick load was evaluated weekly by counting. The MaO1 treatment significantly reduced the tick count only on day +21, reaching approximately 55% efficacy. On the other hand, MaO2 showed significantly lower tick counts on days +7, +14, and +21 after treatment, with weekly efficacy achieving 66%. The results showed a substantial reduction of tick infestation, up to day +28, using a novel formulation of M. anisopliae based in the mixture of two oils. Moreover, we have shown, for the first time, the feasibility of employing formulations of M. anisopliae for large-scale treatment methods, such as a cattle spray race, which in turn, may increase the use and adhesion to biological control tools among farmers.

Octreotide and pasireotide effects on medullary thyroid carcinoma (MTC) cells growth, migration and invasion.

Medullary thyroid carcinoma (MTC) is a rare neuroendocrine neoplasm of the parafollicular thyroid C cells. Although somatostatin receptors are expressed by MTCs, treatment with octreotide has shown poor efficacy, whereas recently pasireotide has demonstrated antiproliferative effects in persistent postoperative MTCs. Aim of this study was to test the effects of octreotide and pasireotide on MTC cells proliferation, cell cycle proteins expression, MAPK activation, apoptosis, calcitonin secretion, migration and invasion in TT cell line as well as in primary MTC cultured cells. Our results showed that both octreotide and pasireotide reduced TT cell proliferation (-35.2 ± 12.1%, p < 0.001, and -25.3 ± 24.8%, p < 0.05, at 10-8 M, respectively), with concomitant inhibition of ERK phosphorylation and cyclin D1 expression. This cytostatic effect was accompanied by a proapoptotic action, with an increase of caspase3/7 activity of 1.5-fold. Moreover, both octreotide and pasireotide inhibited cell migration (-50.9 ± 11.3%, p < 0.01, and -40.5 ± 17%, p < 0.05, respectively) and invasion (-61.3 ± 35.1%, p < 0.05, and -49.7 ± 18%, p < 0.01, respectively). No effect was observed on calcitonin secretion. We then tried to extend these observations to primary cultures (n = 5). Octreotide and/or pasireotide were effective in reducing cells proliferation in 3 out of 5 tumors, and to induce cell apoptosis in 1 out of 3 MTCs. Both octreotide and pasireotide were able to reduce cell migration in all MTC tested. SST2, SST3 and SST5 were expressed in all MTC, with a tendency to increased expression of SST2 in RET mutated vs wild type MTCs. In agreement, inhibition of mutated RET in TT cells reduced SST2 expression. In conclusion, we demonstrated that octreotide and pasireotide inhibited cell proliferation and invasiveness in a subset of MTC, supporting their potential use in the control of tumor growth.

Isolated and combined organic acids in diets of broiler chickens challenged with Eimeria acervulina / [Ácidos orgânicos isolados ou associados em dietas de frangos de corte desafiados com Eimeria acervulina]

This study aimed to evaluate the effect of isolated or combined citric and benzoic acids added to the diets of broiler chickens on performance, allometry of the digestive system organs, intestinal pH and quantity of microorganisms in the jejunum. A total of 840 male Cobb broiler chicks were utilized, distributed in a complete random design in 2 × 2 factorial scheme, supplemented or not with citric acid, and sodium benzoate, with seven replications. At 14 days old, 1mL of a solution containing 1 × 105 sporulated oocysts of Eimeria acervulina per bird was inoculated orally. There was no effect of the acids on the broiler's performance in the 1 to 21-day period. In the total period (1 to 42 days), the broilers fed with a blend of citric and benzoic acid presented greater feed intake, without increment in weight gain. The data of allometry of the digestive system organs and the jejunal pH values were not influenced by the treatments. The citric acid increased the bacterial quantity of gram-positive coccus and total anaerobes in the jejunum. Under the conditions of the present study, we conclude that the citric and benzoic acids, isolated or combined, do not benefit the nutrition of broilers challenged with E. acervulina.(AU)

O objetivo deste trabalho foi avaliar os efeitos da inclusão isolada ou associada dos ácidos cítrico e benzoico na alimentação de frangos de corte sobre o desempenho, a alometria de órgãos do sistema digestório, o pH intestinal e a quantidade de microrganismos no jejuno. Foram utilizados 840 pintos de corte, machos, da linhagem Cobb, distribuídos num delineamento inteiramente ao acaso, em esquema fatorial 2 × 2, com suplementação ou não de ácido cítrico e suplementação ou não de benzoato de sódio, com sete repetições. Aos 14 dias de idade, foi inoculado, via oral, 1mL de solução contendo 1 × 105 oocistos esporulados de Eimeria acervulina por ave. Não houve efeito dos ácidos sobre o desempenho dos frangos no período de um a 21 dias. No período total (um a 42 dias), os frangos alimentados com a mistura de ácidos cítrico e benzoico apresentaram maior consumo de ração, sem incremento no ganho de peso. Os dados de alometria dos órgãos do sistema digestório e os valores de pH do jejuno não foram influenciados pelos tratamentos. O ácido cítrico aumentou a quantidade de bactérias do gênero cocos Gram positivos e anaeróbios totais no jejuno. Nas condições do presente estudo, conclui-se que os ácidos cítrico e benzoico, isolados ou associados, não beneficiam a nutrição de frangos de corte desafiados com E. acervulina.(AU)

Cofilin is a mediator of RET-promoted medullary thyroid carcinoma cell migration, invasion and proliferation.

Medullary thyroid carcinoma (MTC) is a rare neuroendocrine tumor that originates from parafollicular thyroid C cells and accounts for 5% of thyroid cancers. In inherited cases of MTC, and in about 40% of sporadic cases, activating mutations of the receptor tyrosine kinase proto-oncogene RET are found. Constitutively active RET triggers signaling pathways involved in cell proliferation, survival and motility, but the mechanisms underlying malignant transformation of C-cells have been only partially elucidated. Cofilin is a key regulator of actin cytoskeleton dynamics. A crucial role of cofilin in tumor development, progression, invasion and metastasis has been demonstrated in different human cancers, but no data are available in MTC. Interestingly, RET activation upregulates cofilin gene expression. The aim of this study was to investigate cofilin contribution in invasiveness and growth of MTC cells, and its relevance in the context of mutant RET signaling. We found that cofilin transfection in human MTC cell line TT significantly increased migration (178 ±â€¯44%, p < 0.001), invasion (165 ±â€¯28%, p < 0.01) and proliferation (146 ±â€¯18%, p < 0.001), accompanied by an increase of ERK1/2 phosphorylation (2.23-fold) and cyclin D1 levels (1.43-fold). Accordingly, all these responses were significantly reduced after genetic silencing of cofilin (-55 ±â€¯10% migration, p < 0.001, -41 ±â€¯8% invasion, p < 0.001, -17 ±â€¯3% proliferation, p < 0.001). These results have been confirmed in primary cells cultures obtained from human MTCs. The inhibition of constitutively active RET in TT cells by both the RET pharmacological inhibitor RPI-1 and the transfection of dominant negative RET mutant (RETΔTK) resulted in a reduction of cofilin expression (-37 ±â€¯8%, p < 0.001 and -31 ±â€¯16%, p < 0.01, respectively). Furthermore, RPI-1 inhibitory effects on TT cell migration (-57 ±â€¯13%, p < 0.01), but not on cell proliferation, were completely abolished in cells transfected with cofilin. In conclusion, these data indicate that an unbalanced cofilin expression, induced by oncogenic RET, contributes to promote MTC invasiveness and growth, suggesting the possibility of targeting cofilin pathway for more effective treatment of MTC.

Cardiotoxicity and pro-inflammatory effects of the immune checkpoint inhibitor Pembrolizumab associated to Trastuzumab.

BACKGROUND: The immunotherapy has revolutionized the world of oncology in the last decades with considerable advantages in terms of overall survival in cancer patients. The association of Pembrolizumab and Trastuzumab was recently proposed in clinical trials for the treatment of Trastuzumab-resistant advanced HER2-positive breast cancer. Although immunotherapies are frequently associated with a wide spectrum of immune-related adverse events, the cardiac toxicity has not been properly studied. PURPOSE: We studied, for the first time, the putative cardiotoxic and pro-inflammatory effects of Pembrolizumab associated to Trastuzumab. METHODS: Cell viability, intracellular calcium quantification and pro-inflammatory studies (analyses of the production of Interleukin 1ß, 6 and 8, the expression of NF-kB and Leukotriene B4) were performed in human fetal cardiomyocytes. Preclinical studies were also performed in C57BL6 mice by analyzing fibrosis and inflammation in heart tissues. RESULTS: The combination of Pembrolizumab and Trastuzumab leads to an increase of the intracellular calcium overload (of 3 times compared to untreated cells) and to a reduction of the cardiomyocytes viability (of 65 and 20-25%, compared to untreated and Pembrolizumab or Trastuzumab treated cells, respectively) indicating cardiotoxic effects. Notably, combination therapy increases the inflammation of cardiomyocytes by enhancing the expression of NF-kB and Interleukins. Moreover, in preclinical models, the association of Pembrolizumab and Trastuzumab increases the Interleukins expression of 40-50% compared to the single treatments; the expression of NF-kB and Leukotriene B4 was also increased. CONCLUSION: Pembrolizumab associated to Trastuzumab leads to strong cardiac pro-inflammatory effects mediated by overexpression of NF-kB and Leukotriene B4 related pathways.

Hypomorphic Rag1 mutations alter the preimmune repertoire at early stages of lymphoid development.

Hypomorphic RAG1 mutations allowing residual T- and B-cell development have been found in patients presenting with delayed-onset combined immune deficiency with granulomas and/or autoimmunity (CID-G/AI) and abnormalities of the peripheral T- and B-cell repertoire. To examine how hypomorphic Rag1 mutations affect the earliest stages of lymphocyte development, we used CRISPR/Cas9 to generate mouse models with mutations equivalent to those found in patients with CID-G/AI. Immunological characterization showed partial development of T and B lymphocytes, with persistence of naïve cells and preserved serum immunoglobulin but impaired antibody responses and presence of autoantibodies, thereby recapitulating the phenotype seen in patients with CID-G/AI. By using high-throughput sequencing, we identified marked skewing of Igh V and Trb V gene usage in early progenitors, with a bias for productive Igh and Trb rearrangements after selection occurred and increased apoptosis of B-cell progenitors. Rearrangement at the Igk locus was impaired, and polyreactive immunoglobulin M antibodies were detected. This study provides novel insights into how hypomorphic Rag1 mutations alter the primary repertoire of T and B cells, setting the stage for immune dysregulation frequently seen in patients.

Impact of IGF(CA)19 gene polymorphism on the metabolic response to GH therapy in adult GH-deficient patients.

OBJECTIVE: A polymorphism in the promoter region of the IGF1 gene has been linked to serum IGF1 levels, risk of diabetes, and cardiovascular diseases with conflicting results. The aim of this study was to investigate the impact of this polymorphism on the short-term (1 year, n=98) and long-term (5 years, n=50) metabolic response to recombinant human GH (rhGH) in GH-deficient (GHD) adults. DESIGN AND METHODS: Prospective study on GHD adults. Different genotypes were studied by microsatellite method. According to the most frequent 192 bp allele (19 cytosine-adenosine-repeats), subjects were divided into homozygous (19/19), heterozygous (19/X), and noncarriers (X/X). RESULTS: Basal characteristics of patients as well as their response to rhGH in terms of decrease in body fat percentage and increase in IGF1 levels were not different in the three genotype-groups. Conversely, after 1-year rhGH, a significant worsening of insulin sensitivity (i.e. increase in fasting glucose levels and homeostasis model assessment of insulin resistance) and a significant improvement in lipid profile (i.e. reduction in total cholesterol and LDL-cholesterol) were recorded only in homozygous subjects. In the long-term, insulin sensitivity was restored in all the patients, while a significant improvement in lipid profile was observed in homozygous and heterozygous subjects, but not in noncarrier subjects. No difference in rhGH dose among groups was recorded throughout the study. CONCLUSIONS: In GHD adults, the presence of the WT allele in the IGF1 gene promoter may enhance sensitivity to either negative or positive metabolic changes induced by rhGH.

Critical illness in energy metabolism genetic disorder: rhabdomyolysis, acute kidney injury, respiratory arrest / Enfermedad crítica por trastorno genético en el metabolismo de la energía: rabdomiolisis, lesión renal aguda, paro respiratorio

In very long-chain acylCoA dehydrogenase deficiency (VLCAD), the activity of this enzyme is either reduced or absent with the inability to use long-chain fatty acids as energy substrates. A 25-year old male with VLCAD was admitted to the Emergency Department of Policlinico Teaching Hospital (Modena, Italy) for generalized weakness and oliguria, after a period of physical and mental stress and inadequate compliance to a long-chain fatty acid free diet. Laboratory tests were compatible with acute kidney injury. Seventy-two hours after admission, the subject had an episode of chest pain with elevated markers of myocardial necrosis. The rapid deterioration of muscular strength and the subsequent worsening respiratory failure necessitated ventilator support within the local Medical Intensive Care Unit. There, the patient showed a prompt normalization of respiratory parameters and a steady improvement of renal function. An inadequate compliance to lifestyle and dietary restriction in VLCAD may trigger severe and potentially lethal crisis. The in-hospital management of these patients calls for early intensive care admission as their conditions may deteriorate without warning.

En el caso de deficiencia de acylCoA deshidrogenasa de cadena muy larga (VLCAD), o bien se reduce la actividad de esta enzima, o la misma se halla ausente con la consiguiente incapacidad para utilizar los ácidos grasos de cadena larga como sustratos de energía. Un hombre de 25 años fue ingresado con VLCAD fue admitido en el Departamento de emergencia del Hospital Policlínico Docente Hospital de Modena, en Italia, a causa de presentar debilidad generalizada y oliguria, después de un período de estrés físico y mental, y por no cumplir adecuadamente con una dieta libre de ácidos grasos de cadena larga. Las pruebas de laboratorio eran compatibles con una lesión renal aguda. Setenta y dos horas después de su ingreso, el sujeto tuvo un episodio de dolor en el pecho con marcadores elevados de necrosis miocárdica. El rápido deterioro de la fuerza muscular, y el posterior empeoramiento de la insuficiencia respiratoria requirieron el apoyo de un ventilador en la Unidad de Cuidados Médicos Intensivos. Una vez allí, el paciente mostró una pronta normalización de los parámetros respiratorios, y una constante mejoría de la función renal. Un inadecuado cumplimiento con las restricciones dietéticas y el estilo de vida en los casos de VLCAD, pueden desatar una crisis grave y potencialmente fatal. El tratamiento intrahospitalario de estos pacientes requiere un ingreso temprano en cuidados intensivos, ya que sus condiciones pueden deteriorarse sin previo aviso.

Screening for GNAS genetic and epigenetic alterations in progressive osseous heteroplasia: first Italian series.

Progressive osseous heteroplasia (POH) is a rare autosomal dominant disorder of mesenchymal differentiation characterized by progressive heterotopic ossification (HO) of dermis, deep connective tissues and skeletal muscle. Usually, initial bone formation occurs during infancy as primary osteoma cutis (OC) then progressively extending into deep connective tissues and skeletal muscle over childhood. Most cases of POH are caused by paternally inherited inactivating mutations of GNAS gene. Maternally inherited mutations as well as epigenetic defects of the same gene lead to pseudohypoparathyroidism (PHP) and Albright's hereditary osteodystrophy (AHO). During the last decade, some reports documented the existence of patients with POH showing additional features characteristic of AHO such as short stature and brachydactyly, previously thought to occur only in other GNAS-associated disorders. Thus, POH can now be considered as part of a wide spectrum of ectopic bone formation disorders caused by inactivating GNAS mutations. Here, we report genetic and epigenetic analyses of GNAS locus in 10 patients affected with POH or primary OC, further expanding the spectrum of mutations associated with this rare disease and indicating that, unlike PHP, methylation alterations at the same locus are absent or uncommon in this disorder.

Biodegradable core-shell nanoassemblies for the delivery of docetaxel and Zn(II)-phthalocyanine inspired by combination therapy for cancer.

Combination therapies for cancer aim to exploit either additive or synergistic effects arising from the action of two species with the final goal to maximize the therapeutic efficacy. In this work, we develop multifunctional nanoparticles (NPs) for co-delivery of the conventional anticancer drug docetaxel (DTX) and the second generation photosensitizer zinc-phthalocyanine (ZnPc) as potential dual carrier system for the combination of chemotherapy and photodynamic therapy (PDT). Biodegradable and amphiphilic block copolymers based on poly(ε-caprolactone) (PCL=B) and poly(ethylene oxide) (PEO=A), with AB and ABA architectures, were assembled in "core-shell" NPs and loaded with both DTX and ZnPc employing the melting/sonication method. Hydrodynamic diameters within the range 60-100nm and low polydispersity indexes were obtained. Zeta potential was negative for all the formulations and unaffected by drug encapsulation. Concerning drug loading ability of NPs, the entrapment efficiency was related to initial ZnPc/DTX ratio. Steady-stationary and time-resolved emission fluorescence measurements pointed out the embedding of monomeric ZnPc in the NPs, excluding the presence of ZnPc self-supramolecular oligomers. The release of DTX was biphasic whereas ZnPc remained mainly associated with NPs. Singlet oxygen generation was observed when ZnPc-loaded NPs were irradiated at 610nm within a 45min time range, despite that ZnPc was not released in the medium. Stability of NPs in the presence of serum proteins and plasma was excellent and no toxicity toward red blood cells was found. NPs cytotoxicity was evaluated in HeLa cells irradiated for 30min with a halogen lamp. After 72h, viability of cells treated with ZnPc/DTX-loaded NPs strongly decreased as compared to NPs loaded only with DTX, thus showing a combined effect of both DTX and ZnPc. Superior antitumor activity of ZnPc/DTX-loaded NPs as compared to DTX-loaded NPs was confirmed in an animal model of orthotopic amelanotic melanoma, thus pointing to the application of PEO-PCL NPs in the combined chemo-photodynamic therapy of cancer.

BAG3 controls angiogenesis through regulation of ERK phosphorylation.

BAG3 is a co-chaperone of the heat shock protein (Hsp) 70, is expressed in many cell types upon cell stress, however, its expression is constitutive in many tumours. We and others have previously shown that in neoplastic cells BAG3 exerts an anti-apoptotic function thus favoring tumour progression. As a consequence we have proposed BAG3 as a target of antineoplastic therapies. Here we identify a novel role for BAG3 in regulation of neo-angiogenesis and show that its downregulation results in reduced angiogenesis therefore expanding the role of BAG3 as a therapeutical target. In brief we show that BAG3 is expressed in endothelial cells and is essential for the interaction between ERK and its phosphatase DUSP6, as a consequence its removal results in reduced binding of DUSP6 to ERK and sustained ERK phosphorylation that in turn determines increased levels of p21 and p15 and cell-cycle arrest in the G1 phase.

[Lesser validity of urinary benzene than S-phenylmercapturic acid for measuring occupational and environmental exposure to very low concentrations of benzene]. / Minore validità del benzene urinario rispetto all'acido S-fenilmercapturico nel rilevare l'esposizione occupazionale ed ambientale a concentrazioni molto basse di benzene.

AIM: To study the validity of urinary benzene as a biomarker of low and very low exposure to this toxicant, as compared with t,t-muconic acid (t,t-MA) and S-phenylmercapturic acid (SPMA), also taking into account the influence of cigarette smoking and co-exposure to toluene on the urinary excretion of benzene. MATERIALS AND METHODS: The results obtained in two different studies were compared: in the first, workers occupationally exposed to low concentrations of benzene (18 fuel tanker drivers and 23 filling station attendants) were compared with 31 controls and in the second, workers exposed to very low concentrations of benzene (the same 23 filling station attendants) were compared with the 31 controls. Exposure to airborne benzene and toluene was monitored with passive personal samplers (Radiello). Then the urine collected at the end of the work shift was analyzed for t,t-MA, SPMA and urinary benzene. All participants also filled out a questionnaire about their lifestyle habits. RESULTS: There were no differences among the three groups in terms of age and smoking habit. Occupational exposure to benzene and toluene and the urinary concentrations of t,t-MA, SPMA and urinary benzene were higher in the fuel tanker drivers than the filling station attendants and higher in the latter than in the controls. Cigarette smoking was found to be associated with urinary excretion of t,t-MA, SPMA and urinary benzene at both low and very low exposure to benzene. The biomarkers t,t-MA, SPMA and urinary benzene were almost always correlated, for both low and very low exposure to benzene. Notably, for low exposure to benzene a dependency relation was found with the levels of t,t-MA, SPMA and urinary benzene on both cigarette smoking and airborne benzene, whereas for very low exposure to benzene there was a dependency relation of SPMA on cigarette smoking and airborne benzene, of urinary benzene only on cigarette smoking and of t,t-MA on none of the variables considered. CONCLUSIONS: For occupational exposure to low concentrations of benzene, urinary benzene and SPMA showed a comparable validity, while for exposure to very low concentrations of this toxicant the validity of SPMA was confirmed while urinary benzene was found to be less useful. Cigarette smoking was the main factor conditioning the excretion of all the biomarkers of benzene in conditions of both low and very low exposure to the toxicant, so for the analysis of occupational exposure to benzene it is best to recommend abstention from smoking at least in the hours immediately before urine collection.

Applicability of urinary benzene to biological monitoring of occupational and environmental exposure to very low benzene concentrations.

OBJECTIVE: To verify whether urinary benzene is an applicable biomarker of occupational exposure to very low concentrations of benzene, considering the influence of cigarette smoke and benzene-toluene co-exposure. MATERIALS AND METHODS: 23 filling station attendants with occupational exposure to benzene and 31 controls were analyzed. Occupational and environmental exposure was monitored and t,t-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA), urinary benzene and creatinine in the urine samples were tested. RESULTS: Occupational exposure to benzene and toluene was significantly higher in the filling station attendants than in the controls, whereas t,t-MA, SPMA and urinary benzene were not different in the two groups. Instead, the smoker group showed significantly higher values for the above biomarkers than the non-smoker group, each of which included both exposed workers and controls. SPMA was dependent on airborne benzene and cigarette smoking, and urinary benzene only on cigarette smoking, while t,t-MA was not dependent on either of these variables. CONCLUSIONS: At very low concentrations of occupational exposure to benzene, urinary benzene is less valid than SPMA as a biomarker, even if both are strongly influenced by smoking habit. Abstention from smoking should therefore be recommended for at least two hours before urine collection.

[Influence of non-occupational sources on the levels of biomarkers of internal dose for use in biological monitoring of occupational exposure to extremely low concentrations of benzene]. / Ruolo dei fattori non occupazionali nel condizionare i livelli di indicatori di dose interna utilizzati per monitorare l'esposizione occupazionale a concentrazioni molto molto basse di benzene.

OBJECTIVES: To study how traditional (t,t-muconic acid--t,t-MA and S-phenylmercapturic--SPMA) and new (urinary benzene) urinary biomarkers of internal dose can contribute to exclude an occupational source of exposure to extremely low concentrations of benzene, also analyzing the influence that non-occupational sources of exposure, such as cigarette smoking and urban pollution, can have on the levels of these biomarkers. MATERIALS AND METHODS: Assessment was made of 6 workers employed at a groundwater purification plant polluted by benzene (exposed) and 6 administrative clerks employed at the same plant (controls); both groups included smokers and non-smokers. Environmental monitoring (fixed and personal samplings lasting 8 hours) and biological monitoring (determinations of t,t-MA, SPMA, urinary benzene, and urinary creatinine so as to apply suitable adjustments) were performed in exposed workers on 10 successive days, including also rest days (background exposure), and in controls only once. RESULTS: Airborne benzene always resulted lower than the limit of detection of the analytical method in both fixed and personal samplings done on exposed workers and controls during working days, while personal samplings done on exposed workers during rest days showed benzene concentrations even higher than 5 microg/m3, that is the limit value for ambient air quality. Concentrations of t,t-MA, SPMA and urinary benzene did not show differences between exposed workers, regardless of whether they were studied on working or rest days, and controls and appeared to be largely within the reference value range for the Italian population. All biomarkers of internal dose examined in the study showed significantly higher values in smokers than non-smokers. In the latter, SPMA was always below the limit of detection, while urinary benzene resulted higher than the limit of detection in 60.0% and 87.5% of the determinations done on working and on rest days, respectively. CONCLUSIONS: In situations of occupational exposure to extremely low doses of benzene or of absence of exposure, the application of an integrated environmental--biological monitoring approach, involving the determination of SPMA and/or urinary benzene, together with a careful evaluation of those factors determining non-occupational exposure to the toxicant, seems indispensable in order to be able to exclude the presence of occupational exposure. In these particular situations of occupational exposure to benzene, the interpretation of the results of environmental and biological monitoring should not only consider the TLV or BEI, but also the limit value for ambient air quality and the reference value for the general population, since benzene is able to determine genotoxic carcinogenic effects even at exposure to extremely low concentrations of the toxicant.

Perioperative red blood cell transfusion and outcome in stable patients after elective major vascular surgery.

OBJECTIVES: Definitive evidence that red blood cell transfusion improves outcome after vascular surgery is lacking. The aims of the study were to determine, among stable consecutive patients who underwent elective major vascular surgery, (1) the association between postoperative transfusion and 30-day death, myocardial infarction, and both, and (2) and if this association differs according to the presence of postoperative anaemia (haemoglobin value less than 9.0 g/dL within 7 days after surgery). METHODS: A retrospective observational study was conducted on 359 patients prospectively screened according to the ACC/AHA guidelines for preoperative risk in non-cardiac surgery. Main outcome was 30-day death; secondary outcomes 30-day myocardial infarction, and composite of 30-day myocardial infarction or death. RESULTS: Of the patients included, 95 (26.5%) received at least one unit of red blood cells. Patients who received transfusion had a significantly increased hazard of 30-day death (hazard ratio [HR] 11.72, 95% confidence interval [CI] 3.92-35.10; p<0.0001), myocardial infarction (HR 3.3, 95% CI 1.7-6.1; p=0.0003), and both (HR 4.0 95% CI 2.2-7.3; p<0.0001). Such associations held even after adjusting for baseline characteristics, surgical risk, bleeding, and propensity to receive transfusion. There was a significant interaction between transfusion and postoperative anaemia (p=0.012). In patients without anaemia, transfusion was associated with higher risk of 30-day death (HR 19.20, 95% CI 3.99-92.45; p=0.007), myocardial infarction (HR 5.05, 95% CI 2.23-11.44; p=0.0001), and both. Conversely, in patients with anaemia this association was not significant. CONCLUSIONS: In patients who underwent elective major vascular surgery, perioperative transfusion was associated with a significantly increased risk of 30-day events which was more attributable to patients with lesser degree of anaemia. Our data caution against the use of liberal transfusion in stable vascular surgery patients.

[Risk management: the opinions of nursing staff and coordinators in a local health district in the Piemonte region (Italy)]. / Il rischio clinico: opinioni di infermieri e coordinatori dell'ASL 17 della Regione Piemonte.

Clinical risk management includes a set of clinical and administrative activities performed to identify, evaluate and reduce risks for patients, staff and visitors as well as the organization itself. The first fundamental step in risk management is to evaluate risk factors; it is impossible to implement corrective actions and modify and eliminate risk factors if these are not known. The aim of this study was to evaluate the degree to which selected sentinel events were perceived by nursing staff as being severe and whether the degree of perceived severity of an event was associated with specific variables such as nursing work area (medical, surgical, intensive care), years of experience, degree and position. The study also aimed to evaluate the level of knowledge of clinical risk management, identify the main categories of errors within the organization, and evaluate nursing staff opinions on the use of an anonymous system to report errors. A sample of 98 nurses (91 female, 7 male), working in three hospitals in a local health district in the Piemonte region (Italy) participated in the study. The mean age of participants was 37 years (range 22-61). Participants were interviewed between 30 October and 6 November 2006, by using a structured questionnaire. Most participants (93%) were aware of the definition of risk management but over 60% did not use any tool for identifying clinical errors. Nurses perceived infections to be the most serious error, followed by medication errors and surgery and post-operative complications. Almost all participants (99%) considered it right to report errors and 59% admitted to having made errors, most frequently medication errors. Over 90% of participants agreed that an anonymous report form should be used for reporting. Healthcare professionals' willingness to cooperate and their ability to not play down the importance of sentinel events but rather voluntarily bring these to light are essential to the success of risk management in an organization. Clearly, patient safety does not depend on the single individual but rather on an interdisciplinary approach to problem notification and solving and collaboration among interdisciplinary team members.