A new acridone with antifungal properties against Candida spp. and dermatophytes, and antibiofilm activity against C. albicans.

AIM: The increase in the number of fungal infections worldwide, coupled with the limitations of current antifungal chemotherapy, demand the development of safe and effective new antifungals. Here, we presented the synthesis of a novel acridone (M14) and its antifungal properties against Candida and dermatophytes species. METHODS AND RESULTS: A series of 17 acridones was designed, synthesized and tested for its antifungal activity. The minimum inhibitory concentration (MIC) was determined by the broth microdilution method. Only the acridone M14 showed growth-inhibitory activity against reference strains and clinical isolates of Candida and dermatophytes, with MIC range of 7·81-31·25 µg ml-1 . Moreover, M14 exhibited fungicidal activity and prevented biofilm formation by C. albicans as well as reduced the viability of preformed biofilms, even at sub-MICs. The confocal laser scanning microscopy analysis revealed that C. albicans hyphal growth was completely inhibited in the presence of M14. Similarly, there was a severe inhibition on hyphal growth of Trichophyton rubrum. We also found that M14 has relatively low toxicity to human fibroblasts. CONCLUSIONS: The new acridone M14 has antifungal properties against Candida spp. and dermatophytes, and antibiofilm activity against C. albicans. In addition, M14 is relatively selective to fungal cells compared to human normal cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its in vitro antifungal activity, anti-Candida biofilm effect and moderate cytotoxicity towards normal human cell, M14 may serve as a valuable lead compound to develop a new antifungal agent.

Integral nutritional approach to the care of cancer patients: results from a Delphi panel

Introduction: Malnutrition is a common complication in cancer patients and can negatively affect the outcome of treatments. This study aimed to reach a consensus on nutritional needs and optimize nutritional care in the management of cancer patients at a national level. Methods: A qualitative, multicenter, two-round Delphi study involving 52 specialists with experience in nutritional support in cancer patients was conducted. Results: Regarding the presence of malnutrition, 57.7% of the participants stated that < 30% of the patients had malnutrition at the time of diagnosis, 40.4% considered that 31-50% had malnutrition during cancer treatment, and 26.9% that > 50% at the end of the treatment. Forty percent of participants believed that the main objective of nutritional treatment was to improve quality of life and 34.6% to improve tolerability and adherence to chemotherapy. The quality nutritional care provided at their centers was rated as medium-low by 67.3%. Enteral and parenteral nutrition was administered to less than 10% and less than 5% of patients in 40.4 and 76.9% of cases, respectively. In relation to nutritional screening at the time of diagnosis, 62.9% of participants considered than screening to assess the risk of malnutrition was performed in < 30% of patients. Conclusions: There is an important variability in the management of cancer patient nutrition, which is associated with the absence of a national consensus on nutritional support in this field. Given the incidence of nutritional disorders in cancer patients, a specialist in clinical nutrition (regardless of his/her specialty) should be integrated into the strategic cancer plan

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Integral nutritional approach to the care of cancer patients: results from a Delphi panel.

INTRODUCTION: Malnutrition is a common complication in cancer patients and can negatively affect the outcome of treatments. This study aimed to reach a consensus on nutritional needs and optimize nutritional care in the management of cancer patients at a national level. METHODS: A qualitative, multicenter, two-round Delphi study involving 52 specialists with experience in nutritional support in cancer patients was conducted. RESULTS: Regarding the presence of malnutrition, 57.7% of the participants stated that < 30% of the patients had malnutrition at the time of diagnosis, 40.4% considered that 31-50% had malnutrition during cancer treatment, and 26.9% that > 50% at the end of the treatment. Forty percent of participants believed that the main objective of nutritional treatment was to improve quality of life and 34.6% to improve tolerability and adherence to chemotherapy. The quality nutritional care provided at their centers was rated as medium-low by 67.3%. Enteral and parenteral nutrition was administered to less than 10% and less than 5% of patients in 40.4 and 76.9% of cases, respectively. In relation to nutritional screening at the time of diagnosis, 62.9% of participants considered than screening to assess the risk of malnutrition was performed in < 30% of patients. CONCLUSIONS: There is an important variability in the management of cancer patient nutrition, which is associated with the absence of a national consensus on nutritional support in this field. Given the incidence of nutritional disorders in cancer patients, a specialist in clinical nutrition (regardless of his/her specialty) should be integrated into the strategic cancer plan.

Beneficial effects of acute trans-resveratrol supplementation in treated hypertensive patients with endothelial dysfunction.

Endothelial dysfunction is a surrogate marker of cardiovascular risk. Resveratrol is known to improve endothelial function in animals, however, clinical trials are limited. We hypothesized that the acute trans-resveratrol supplementation improves endothelial function in treated hypertensive patients with endothelial dysfunction. Twenty-four hypertensive patients between 45 and 65 years-old with baseline endothelial dysfunction were enrolled in a randomized, cross-over, double-blind, placebo-controlled trial. Individuals received either a single dose of trans-resveratrol (300 mg) or placebo and were crossed-over after a one-week washout period. Blood pressure (BP) measurements, aortic systolic blood pressure (SBP) and brachial flow-mediated dilation (FMD) were performed before and 1.5 hours after the intervention. FMD was significantly increased in women (4.2 ± 0.5 vs 7.1 ± 1.3%, p = 0.026) but not in men (4.4 ± 0.9 vs 4.9 ± 0.8%, p = 0.588) in the trans-resveratrol group. There was no statistical difference between baseline and final values of brachial BP and also no changes in aortic SBP. Patients with higher low-density lipoprotein (LDL) cholesterol had better FMD response to trans-resveratrol than patients with lower LDL cholesterol (7.4 ± 1.2 vs 4.3 ± 1.0%, p = 0.004). Our study demonstrated that the acute supplementation of trans-resveratrol promoted an improvement in endothelial function, especially in women and those with higher LDL-cholesterol, despite no changes in BP. List of Abbreviation: Aix: augmentation index; AP: augmentation pressure; BP: blood pressure; BMI: body Mass Index; CVD: cardiovascular disease; FMD: flow-mediated dilation; FRS: Framingham Risk Score; HDL: high-density lipoprotein; LDL: low-density lipoprotein; NO: nitric oxide; SPSS: Statistical Package for Social Sciences; ROS: reactive oxygen species; SBP: systolic blood pressure; TG: triglycerides.

Lipocalin-2 regulates adult neurogenesis and contextual discriminative behaviours.

In the adult mammalian brain, newborn granule cells are continuously integrated into hippocampal circuits, and the fine-tuning of this process is important for hippocampal function. Thus, the identification of factors that control adult neural stem cells (NSCs) maintenance, differentiation and integration is essential. Here we show that the deletion of the iron trafficking protein lipocalin-2 (LCN2) induces deficits in NSCs proliferation and commitment, with impact on the hippocampal-dependent contextual fear discriminative task. Mice deficient in LCN2 present an increase in the NSCs population, as a consequence of a G0/G1 cell cycle arrest induced by increased endogenous oxidative stress. Of notice, supplementation with the iron-chelating agent deferoxamine rescues NSCs oxidative stress, promotes cell cycle progression and improves contextual fear conditioning. LCN2 is, therefore, a novel key modulator of neurogenesis that, through iron, controls NSCs cell cycle progression and death, self-renewal, proliferation and differentiation and, ultimately, hippocampal function.

Physics, chemistry, and Hirshfeld surface analyses of gamma-irradiated thalidomide to evaluate behavior under sterilization doses / 药物分析学报

Thalidomide was indicated as a sedative and antiemetic and prescribed for pregnant women. Its tragic teratogenic effects culminated in withdrawal from the market. Since the discovery of its anti-angiogenic and anti-inflammatory actions, thalidomide has been used in the treatment of leprosy and multiple myeloma, which justify studies of its stability. We investigated the effects of irradiation of thalidomide up to 100 kGy (fourfold the usual sterilizing dose for pharmaceutics). The β polymorph of thalidomide was obtained in an isothermal experiment at 270 °C. All samples underwent gamma irradiation for specific times. At different doses, decomposition of the pharmaceutical was not observed up to 100 kGy. The observed effect was angle turning between the phthalimide and glutarimide rings modulated by repulsion towards the carbonyl group, leading to a stable energetic configuration, as measured by the equilibrium in the torsion angle after irra-diation. The thalidomide molecule has a center of symmetry, so a full turn starting from 57.3° will lead to an identical molecule. Further irradiation will start the process again. Samples irradiated at 30 and 100 kGy have more compact unit cells and a lower volume, which leads to an increase in the intermolecular hydrogen interaction within the unit cel , resulting in higher thermal stability for polymorph α.

Physicochemical characterization, the Hirshfeld surface, and biological evaluation of two meloxicam compounding pharmacy samples / 药物分析学报

Meloxicam (MLX) is an anti-inflammatory drug susceptible to variations and crystalline transitions. In compounding pharmacies, the complete crystallographic evaluation of the raw material is not a routine procedure. We performed a complete crystallographic characterization of aleatory raw MLX samples from compounding pharmacies. X-ray diffraction indicated the presence of two crystalline forms in one sample. DSC experiments suggested that crystallization, or a crystal transition, occurred differently be-tween samples. The FTIR and 1H NMR spectra showed characteristic assignments. 13C solid-state NMR spectroscopy indicated the presence of more than one phase in a sample from pharmacy B. The Hirshfeld surface analysis, with electrostatic potential projection, allowed complete assignment of the UV spectra in ethanol solution. The polymorph I of meloxicam was more active than polymorph Ⅲ in an experi-mental model of acute inflammation in mice. Our results highlighted the need for complete crystal-lographic characterization and the separation of freely used raw materials in compounding pharmacies, as a routine procedure, to ensure the desired dose/effect.

Very late presentation of a disorder of sex development.

Disorders of sex development generally present in the neonatal period with ambiguity of external genitalia. We report a very old male patient presenting at 75 years because of panhypopituitarism and a large nonsecreting pituitary macroadenoma secondary to long-standing primary hypogonadism due to 46,XX sex reversal disorder now first diagnosed. Sex development disorders may go unrecognised for the entire life span, despite infertility and long-standing primary gonadic failure may lead to uncommon complications.

Challenges for the WFD second management cycle after the implementation of a regional multi-municipality sanitation system in a coastal lagoon (Ria de Aveiro, Portugal).

In the scope of the first WDF management cycle, a multi-municipality sanitation system with secondary treatment was implemented in 2005 in Ria de Aveiro coastal lagoon, with the treated effluent discharging into the Atlantic Ocean through a submarine outfall. The lagoon water chemical status was evaluated regarding dissolved inorganic nutrients and chlorophyll a before and after 2005. The S. Jacinto outfall has effectively reduced the point source nutrient loads (ammonium and phosphate) into Ria de Aveiro, representing a step forward for the implementation of the WFD, through eutrophication abatement. However, the lagoon remains exposed to non-point nitrogen sources, as a consequence of the current land use and water management, which in a scenario of winter extreme precipitation events, nutrients increase through surface run-off. Besides, in a combined scenario of low freshwater input into the lagoon and favourable oceanic condition, nutrients enter through Ria outer boundary coming from the S. Jacinto outfall. Thus, changes in the system hydrodynamics in the context of global change might pose new challenges regarding the WFD second management cycle involving the second river basin management plan and the first flood risk management plan, foreseeing the 'Good' ecological status in all Ria's Water Bodies.

Visceral adiposity is associated with cytokines and decrease in lung function in women with persistent asthma.

UNLABELLED: Abdominal obesity is associated with a risk of cardiovascular diseases, metabolic syndrome and decreased lung function. However, it is not known whether asthma control is influenced by the accumulation of adipose tissue in the various abdominal compartments. OBJECTIVE: To determine associations among abdominal adiposity distribution, asthma control, lung function and cytokines in women. METHODS AND DESIGN: In this cross-sectional study of asthmatic women, data on demographic variables, comorbid conditions, disease history, anthropometric and spirometric measurements were collected. Subcutaneous (SAT) and visceral (VAT) adipose tissues were measured by ultrasound, and the steatosis level was obtained. Asthma control was assessed according to Global Initiative for Asthma (GINA) criteria. Atopy was defined on the basis of allergen-specific Immunoglobulin E and/or skin prick testing. Cytokine levels were determined using enzyme-linked immunosorbant assays (ELISAs). RESULTS: Eighty-three asthmatic women were included, 37% of whom had uncontrolled asthma. After controlling for variables, a negative association between asthma control and VAT and the VAT/SAT ratio was observed. VAT was negatively associated with respiratory parameters after controlling for explanatory variables. In an adjusted model, body mass index (BMI) and SAT were inversely associated with the adiponectin serum level and VAT was associated with the interleukin 6 level. In conclusion, visceral obesity was negatively associated with asthma control and lung function; and positively associated with increased levels of interleukin 6 in women. We hypothesize that women should be studied as a separate group, and we suggest further studies with a control group to know if the uncontrolled asthmatic group is directly affected by visceral adipose inflammatory markers.

Alivio del dolor y el estrés al vacunar. Síntesis de la evidencia. Recomendaciones del Comité Asesor de Vacunas de la AEP / Relief of pain and distress during immunizations. Synthesis of evidence. Recommendations of the Advisory Committee on Vaccines of the Spanish Association of Paediatrics

Introducción: en niños y adolescentes sanos, las vacunaciones son con frecuencia fuente de dolor y sufrimiento. Padres, niños, adolescentes y profesionales sanitarios muestran preocupación sobre ello. El Comité Asesor de Vacunas de la Asociación Española de Pediatría (CAV-AEP) cree que abordar el dolor y el sufrimiento al vacunar es necesario, siguiendo la metodología de la medicina basada en la evidencia. El objetivo del presente trabajo es elaborar recomendaciones basadas en el conocimiento científico. Material y métodos: se dividió la materia de estudio en cuatro áreas: amamantamiento y soluciones azucaradas, anestésicos tópicos, métodos para la administración de vacunas y otras intervenciones (distracción). Se realizó una síntesis de la evidencia, asumiendo las recomendaciones de la Guía de práctica clínica de Anna Taddio (2010) e incorporando la evidencia de revisiones sistemáticas y ensayos clínicos posteriores a los incorporados en dicha guía. Resultados: las medidas que se han mostrado efectivas en la disminución del dolor han sido las siguientes: en lactantes, amamantar antes, durante y después de la inyección; las soluciones azucaradas son una alternativa si la lactancia materna no fuera posible; los anestésicos tópicos son eficaces para todas las edades, pero requieren un tiempo para mostrar su efecto y tienen un coste; no aspirar en la inyección intramuscular y hacerlo lo más rápido posible; administrar las vacunas de forma que la más dolorosa sea la última; cuando sea posible, es preferible inyectar simultáneamente más de una vacuna que hacerlo de forma secuencial; sostener al niño en brazos; y utilizar maniobras de distracción para niños de 2-14 años. Conclusiones: realizada una exhaustiva revisión del tema, hay pruebas suficientes para afirmar que los profesionales que administran vacunas infantiles deberían poner en práctica medidas para atenuar el dolor que indudablemente acompaña al procedimiento de la vacunación. Se trata además, en general, de medidas técnicamente sencillas y fáciles de incorporar a la práctica (AU)

Background: in healthy children and adolescents, immunizations that require a needle related procedure are the most common source of pain and distress. Parents, children, adolescents and health-care providers are concerned about this. The Advisory Committee on Immunization of the Spanish Association of Pediatrics (CAV-AEP) believes that address pain and distress at the time of vaccination is necessary following recommendations that have to be based on rigor and science. Methods: we divided the subject in four areas: Breastfeeding and oral sucrose solutions, topical anaesthetics, vaccination administration methods and other interventions (distraction). Synthesis of evidence was made. Assuming the recommendations of The Clinical Guideline of Anne Taddio (2010) and adding the evidence of clinical trial published after the Guide. Results: methods that showed effectiveness in diminishing pain were: for infants, breastfeeding before, during and after the puncture is effective in manage pain. Oral sucrose solutions could be an alternative if breastfeeding is not possible. Topical anaesthetics are effective for all ages but a time to produce effect is required and need financial resources. No aspiration for intramuscular injection, put the injection as quickly as possible, give the vaccines so that the most painful the last. If more than one vaccine injection are required in the same visit, and it is possible, it is preferable to inject simultaneously more than one vaccine than sequentially. Hold the infant. For children 2 to 14 years use distraction techniques. Conclusions: as a thorough revision of the topic was made, there are enough evidence to recommend that in any setting where children immunization is given, techniques to mitigate pain at the time of vaccination should be implemented, moreover these strategies are simple and easy to assimilate in clinical practice (AU)

Experimental simulation of decoherence in photonics qudits.

We experimentally perform the simulation of open quantum dynamics in single-qudit systems. Using a spatial light modulator as a dissipative optical device, we implement dissipative-dynamical maps onto qudits encoded in the transverse momentum of spontaneous parametric down-converted photon pairs. We show a well-controlled technique to prepare entangled qudits states as well as to implement dissipative local measurements; the latter realize two specific dynamics: dephasing and amplitude damping. Our work represents a new analogy-dynamical experiment for simulating an open quantum system.

Llanto al mes de vida. No todo son cólicos / Crying in a one-month-old baby. Not all are crampings

El llanto de los lactantes es uno de los motivos de consulta más frecuentes en las Urgencias Pediátricas. La mayoría de las ocasiones son cuadros autolimitados, pero hay que prestar especial atención a los signos y síntomas de alarma para realizar otras pruebas complementarias. Presentamos el caso de un lactante de un mes de vida que asociaba distensión abdominal y rechazo de las tomas. En las pruebas de imagen se apreció una masa multiquística en abdomen. La laparotomía evidenció una malformación linfática en el íleon medio, con confirmación anatomopatológica de linfagioma quístico mesentérico (AU)

Crying infants are one of the most frequent reasons within the pediatric emergency consultation. Most cases are self-limiting but we must pay special attention to the alarm signs/symptoms to perform additional tests. We report the case of a one-month-old infant who presents bloating and rejects feeding. The imaging test shows a multicystic mass in the abdomen. Laparotomy revealed a lymphatic malformation in the middle ileum and pathology confirmed mesenteric cystic lymphangioma (AU)

Hiperplasia suprarrenal congénita debida a deficiencia de 17 -hidroxilasa: a propósito de una nueva mutación en el gen CYP17A1 / Congenital adrenal hyperplasia due to lack of 17 -hydroxylase: A report of a new mutation in the gene CYP17A1

La enzima P450c17 cataliza 2 reacciones diferentes: 17alfa-hidroxilación de la progesterona y pregnenolona y segmentación de la unión del carbono 17-20 a partir de la 17,20 liasa para producir andrógenos suprarrenales. Esta enzima está codificada por el gen CYP17A1. Se presenta una paciente de 14 años con retraso en el desarrollo puberal y presión arterial elevada para su talla y edad. Cariotipo 46,XX. En el estudio hormonal destaca hipogonadismo hipergonadotropo, así como una insuficiencia suprarrenal y exceso mineralocorticoideo. El estudio genético mostró una mutación en homozigosis en el gen CYP17A1 (c.753+1G>A), no descrita previamente, la cual es responsable de la fisiopatología de la deficiencia de 17alfa-hidroxilasa. Esta entidad es una forma rara de hiperplasia suprarrenal congénita. Normalmente la enfermedad suele pasar desapercibida hasta la adolescencia o el inicio de la vida adulta y se debería sospechar ante individuos 46,XY con genitales ambiguos o 46,XX con retraso puberal que asocia hipertensión y/o hipopotasemia

P450c17 enzyme catalyses two different reactions: the 17Alpha-hydroxylation of progesterone and pregnenolone, and segmenting the carbon 17-20 binding from the 17,20 lyase producing adrenal androgens. This enzyme is coded by the CYP17A1 gene. The case is presented of a 14 year old patient with delayed pubertal development and a high blood pressure for height and age. 46,XX karyotype. Hormonal studies highlighted hypergonadotropic hypogonadism, adrenal insufficiency and mineralocorticoid excess. Subsequent genetic studies showed a homozygous mutation in the CYP17A1 gene (c.753+G>A), not previously described, which is responsible for the pathophysiology of 17Alpha-hydroxylase deficiency. This entity is a rare form of congenital adrenal hyperplasia. The disease often goes unnoticed until adolescence or early adult life, and should be suspected in 46,XY individuals with ambiguous genitalia or 46,XX with delayed puberty associated with hypertension and/or hypokalaemia

Rectangular ion funnel: a new ion funnel interface for structures for lossless ion manipulations.

Structures for lossless ion manipulations (SLIM) have recently demonstrated the ability for near lossless ion focusing, transfer, and trapping in subatmospheric pressure regions. While lossless ion manipulations are advantageously applied to the applications of ion mobility separations and gas phase reactions, ion introduction through ring electrode ion funnels or more conventional ion optics to SLIM can involve discontinuities in electric fields or other perturbations that result in ion losses. In this work, we developed and investigated a new funnel design that aims to seamlessly couple to SLIM at the funnel exit. This rectangular ion funnel (RIF) was initially evaluated by ion simulations, fabricated utilizing printed circuit board technology, and tested experimentally. The RIF was integrated to a SLIM-time of flight (TOF) MS system, and the operating parameters, including RF, DC bias of the RIF electrodes, and electric fields for effectively interfacing with a SLIM, were characterized. The RIF provided a 2-fold sensitivity increase without significant discrimination over a wide m/z range and well matched to that of SLIM, along with greatly improved SLIM operational stability.

Influence of segmental chromosome abnormalities on survival in children over the age of 12 months with unresectable localised peripheral neuroblastic tumours without MYCN amplification.

BACKGROUND: The prognostic impact of segmental chromosome alterations (SCAs) in children older than 1 year, diagnosed with localised unresectable neuroblastoma (NB) without MYCN amplification enrolled in the European Unresectable Neuroblastoma (EUNB) protocol is still to be clarified, while, for other group of patients, the presence of SCAs is associated with poor prognosis. METHODS: To understand the role of SCAs we performed multilocus/pangenomic analysis of 98 tumour samples from patients enrolled in the EUNB protocol. RESULTS: Age at diagnosis was categorised into two groups using 18 months as the age cutoff. Significant difference in the presence of SCAs was seen in tumours of patients between 12 and 18 months and over 18 months of age at diagnosis, respectively (P=0.04). A significant correlation (P=0.03) was observed between number of SCAs per tumour and age. Event-free (EFS) and overall survival (OS) were calculated in both age groups, according to both the presence and number of SCAs. In older patients, a poorer survival was associated with the presence of SCAs (EFS=46% vs 75%, P=0.023; OS=66.8% vs 100%, P=0.003). Moreover, OS of older patients inversely correlated with number of SCAs (P=0.002). Finally, SCAs provided additional prognostic information beyond histoprognosis, as their presence was associated with poorer OS in patients over 18 months with unfavourable International Neuroblastoma Pathology Classification (INPC) histopathology (P=0.018). CONCLUSIONS: The presence of SCAs is a negative prognostic marker that impairs outcome of patients over the age of 18 months with localised unresectable NB without MYCN amplification, especially when more than one SCA is present. Moreover, in older patients with unfavourable INPC tumour histoprognosis, the presence of SCAs significantly affects OS.

Cerebellar vermis H2 receptors mediate fear memory consolidation in mice.

Histaminergic fibers are present in the molecular and granular layers of the cerebellum and have a high density in the vermis and flocullus. Evidence supports that the cerebellar histaminergic system is involved in memory consolidation. Our recent study showed that histamine injections facilitate the retention of an inhibitory avoidance task, which was abolished by pretreatment with an H2 receptor antagonist. In the present study, we investigated the effects of intracerebellar post training injections of H1 and H2 receptor antagonists as well as the selective H2 receptor agonist on fear memory consolidation. The cerebellar vermi of male mice were implanted with guide cannulae, and after three days of recovery, the inhibitory avoidance test was performed. Immediately after a training session, animals received a microinjection of the following histaminergic drugs: experiment 1, saline or chlorpheniramine (0.016, 0.052 or 0.16 nmol); experiment 2, saline or ranitidine (0.57, 2.85 or 5.07 nmol); and experiment 3, saline or dimaprit (1, 2 or 4 nmol). Twenty-four hours later, a retention test was performed. The data were analyzed using one-way analysis of variance (ANOVA) and Duncan's tests. Animals microinjected with chlorpheniramine did not show any behavioral effects at the doses that we used. Intra-cerebellar injection of the H2 receptor antagonist ranitidine inhibited, while the selective H2 receptor agonist dimaprit facilitated, memory consolidation, suggesting that H2 receptors mediate memory consolidation in the inhibitory avoidance task in mice.

[Congenital adrenal hyperplasia due to lack of 17α-hydroxylase: a report of a new mutation in the gene CYP17A1]. / Hiperplasia suprarrenal congénita debida a deficiencia de 17α-hidroxilasa: a propósito de una nueva mutación en el gen CYP17A1.

P450c17 enzyme catalyses two different reactions: the 17α-hydroxylation of progesterone and pregnenolone, and segmenting the carbon 17-20 binding from the 17,20lyase producing adrenal androgens. This enzyme is coded by the CYP17A1 gene. The case is presented of a 14 year old patient with delayed pubertal development and a high blood pressure for height and age. 46,XX karyotype. Hormonal studies highlighted hypergonadotropic hypogonadism, adrenal insufficiency and mineralocorticoid excess. Subsequent genetic studies showed a homozygous mutation in the CYP17A1 gene (c.753+G>A), not previously described, which is responsible for the pathophysiology of 17α-hydroxylase deficiency. This entity is a rare form of congenital adrenal hyperplasia. The disease often goes unnoticed until adolescence or early adult life, and should be suspected in 46,XY individuals with ambiguous genitalia or 46,XX with delayed puberty associated with hypertension and/or hypokalaemia.

Simultaneous detection of hepatitis C virus antigen and antibodies in dried blood spots.

BACKGROUND: Enzyme immunoassays (EIA) designed to detect hepatitis C virus (HCV) core antigen and anti-HCV antibodies (HCV AgAb) simultaneously can improve the early detection of HCV infection when molecular diagnostic methods are not widely available. OBJECTIVES: To evaluate the suitability of dried blood spot (DBS) samples for detecting HCV AgAb using commercial EIAs. STUDY DESIGN: Paired serum and DBS samples were assayed using two commercial EIAs for HCV AgAb (Monolisa™ HCV AgAb ULTRA and Murex HCV AgAb). Manufacturer's recommendations were followed for sera while sample volume, incubation time and cut-off (CO) determination were evaluated for the DBS samples. The values of sensitivity, specificity, inter-rater agreement, detection limit, assay precision and stability of DBS samples at different conditions (22-26°C, 2-8°C and -20°C) were determined. RESULTS: It was necessary to increase the DBS sample volume fourfold compared to the sera samples to approximate the DBS Optical Density (OD) values to the sera OD values. Using ROC curve to recalculate CO values for the DBS samples, sensitivity was 97.5% for both EIAs, while the specificity was 99.71% for Monolisa™ HCV AgAb ULTRA and 95.95% for Murex HCV AgAb. Accurate testing results were obtained with DBS samples for 60 days at all conditions evaluated; storage at -20°C resulted in low OD variation. Both EIAs demonstrated the same limit of detection among DBS samples [estimated viral load of 3.1 International Units per millilitre (IU/mL)] and low OD value variability in repetitivity and reproducibility studies. CONCLUSION: DBS samples can be used for the detection of HCV AgAb by EIA as they present comparable performance characteristics and excellent stability among various storage conditions.