RESUMEN
BACKGROUND: Early detection of developmental delay (DD) in preschool children is crucial for counselling parents, initiating diagnostic work-up, and starting early intervention (EI). METHODS: We conducted a register study of all preschool children referred for EI in the Canton of Zurich, Switzerland, in 2017 (N = 1,785) and used an online survey among primary care physicians (PCPs, N = 271) to evaluate the care service of DD children. RESULTS: PCPs accounted for 79.5% of all referrals by physicians and had correctly referred over 90% of the children in need of EI at an average age of 39.3 months (SD 8.9). In the survey, which represents 59.2% of all pediatricians and 11.3% of all general practitioners in the Canton, PCPs reported performing a mean of 13.5 (range 0-50, SD 10.7) well-child visits per week to preschool children and estimated well-child visits to be the most frequent type of consultation (66.7%) for the identification of DD. Parents' hesitancy in accepting further evaluation or support were reported by 88.7%. CONCLUSIONS: Most preschool children with DD are identified in well-child visits. These visits represent an ideal opportunity for early detection of developmental impairment and initiation of EI. Carefully addressing parents' reservations could reduce the rate of refusal, thus improving early support for children with DD.
Asunto(s)
Médicos Generales , Padres , Humanos , Preescolar , Niño , Examen Físico , Encuestas y Cuestionarios , Pediatras , Discapacidades del Desarrollo/diagnósticoRESUMEN
During the last few years, an important element in the improvement of the molecular biology techniques has been the necessity for availability of high quality and functionality DNA. Several DNA extraction procedures with different results in both performance and quality, have been proposed. In this study our objective was to determine the most reliable extraction method that balances DNA quantity, and to assess the sample quantification of the fluorometric DNA quantification methods. For this, blood extracted by venopunction from 20 healthy volunteers was used to obtain DNA from buffy coat, and 4 commercial DNA extraction kits were assessed as well as two fluorometric DNA quantification methods with protocols of different complexity. Results suggest that manual methods achieve higher quality and larger yields of DNA. DNA purity obtained with the 4 extraction kits evaluated through the 260/280 and 260/230 ratio showed that the Qiacube kit fulfilled the criteria established in this work, followed very close by the Flexigene kit. On the other hand, the fluorometric DNA methods used in the samples quantification showed a higher variability when using QuantiFlour method, obtaining better results probably due to the simplicity of this protocol.
Asunto(s)
Capa Leucocitaria de la Sangre , ADN , Humanos , ADN/aislamiento & purificaciónRESUMEN
This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)
O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)
Asunto(s)
Animales , Cíclidos/crecimiento & desarrollo , Hepatopáncreas/química , Lactobacillus plantarum , Microbioma Gastrointestinal , Alimentación Animal , Factor I del Crecimiento Similar a la Insulina , Suplementos Dietéticos , LeptinaRESUMEN
While the health effects of trypanosomes in Australian mammals in their native range are not fully understood, there is evidence of an impact in those species introduced to other geographical regions. Here we report the pathological and molecular features of concurrent fatal trypanosomiasis and toxoplasmosis in an adult female captive red-necked wallaby (syn. Bennett's wallaby; Macropus rufogriseus) from Bee County, Texas, USA. The animal exhibited no clinical signs prior to sudden death. On necropsy, the main findings were generalized organ congestion and bilateral renal petechiation. Microscopically, the main finding was lymphohistiocytic and necrotizing pancarditis with intrasarcoplasmic protozoal pseudocysts containing amastigotes and occasional intrahistiocytic amastigotes, morphologically compatible with Trypanosoma cruzi, as well as rare intrasarcoplasmic protozoal tissue cysts with zoites morphologically compatible with Toxoplasma gondii. Other lesions included acute centrilobular to panlobular necrotizing hepatitis with intrahepatocellular T. gondii cysts, necrotizing splenitis, pulmonary oedema with fibrin, histiocytosis and rare fibrin microthrombi, and acute renal tubular degeneration with proteinosis and pigmented casts suggestive of haemoglobinuria or myoglobinuria. Immunohistochemical labelling confirmed intralesional T. gondii cysts and molecular analyses identified T. cruzi genotype I and T. gondii. This is a unique case that, to the best of our knowledge, represents the first description of T. cruzi and T. gondii co-infection, as well as the first record of naturally occurring infection T. cruzi genotype I infection in macropodids. This case adds to the epidemiological knowledge on Chagas disease in the USA, particularly in Texas where there is a high prevalence of human and canine trypanosomiasis.
Asunto(s)
Enfermedad de Chagas/veterinaria , Coinfección/veterinaria , Macropodidae , Toxoplasmosis Animal , Animales , FemeninoRESUMEN
Human samples are commonly collected and long-term stored in biobanks for current and future analyses. Even though techniques for freezing human blood are well established, the storage time can compromise the cell viability as well as the yield and quality of nucleic acids (RNA and DNA) extracted from them. In this study, a protocol to obtain peripheral blood mononuclear cells (PBMCs) from 70 subjects, which were stored at - 196 °C from EDTA tubes for a long-term, was assessed. In parallel; a protocol to obtain DNA from the same subjects, which were stored at - 80 °C from citrate tubes, was also studied. Samples stored from 2008 to 2012 were studied and the results obtained showed that there were no statistically significant differences in the RNA or DNA extracted in terms of purity, integrity and functionality The freezing protocol used by the Málaga Biobank shows that viable PBMCs and DNA could be kept for a period of, at least, 10 years, with a high quality and performance. Furthermore, RNA extracted from these PBMCs presents also a good quality and performance. Therefore, the samples frozen according to the conditions of the protocols assessed in this study could be optimal for biomedical research.
Asunto(s)
Criopreservación/métodos , ADN/análisis , Leucocitos Mononucleares/citología , ARN/análisis , Bancos de Sangre , Supervivencia Celular , Humanos , España , Bancos de TejidosRESUMEN
Growth hormone releasing peptides (GHRPs) could be widely used by cheating athletes because they produce growth hormone (GH) secretion, so may generate an ergogenic effect in the body. Knowledge of the essential amino acids needed in GHRP structure for interaction with the target biological receptor GHSR1a, the absorption through different administration routes, and the maintenance of pharmacological activity of potential biotransformation products may help in the fight against their abuse in sport. Several GHRPs and truncated analogues with the common core Ala-Trp-(D-Phe)-Lys have been studied with a radio-competitive assay for the GHSR1a receptor against the radioactive natural ligand ghrelin. Relevant chemical modifications influencing the activity for positions 1, 2, 3, and 7 based on the structure aa-aa-aa-Ala-Trp-(D-Phe)-Lys have been obtained. To test in vivo the applicability of the activities observed, the receptor assay activity in samples from excretion studies performed after nasal administration of GHRP-1, GHRP-2, GHRP-6, Hexarelin, and Ipamorelin was confirmed. Overall results obtained allow to infer structure-activity information for those GHRPs and to detect GHSR1a binding (intact GHRPs plus active metabolites) in excreted urines. Copyright © 2016 John Wiley & Sons, Ltd.
Asunto(s)
Sustancias de Crecimiento/farmacología , Oligopéptidos/farmacología , Receptores de Ghrelina/metabolismo , Doping en los Deportes , Sustancias de Crecimiento/administración & dosificación , Sustancias de Crecimiento/química , Sustancias de Crecimiento/orina , Células HEK293 , Humanos , Masculino , Oligopéptidos/administración & dosificación , Oligopéptidos/química , Oligopéptidos/orina , Relación Estructura-ActividadRESUMEN
Since the appearance of recombinant human growth hormone (rhGH) in the 1980s, its expansion and acquisition through the black market has increased, so the detection of its abuse continues to be a challenge. New biomarkers that are more reliable and sensitive, allowing a larger detection window, are still needed. In this line, Fibronectin 1 (FN1) has been proposed as a potential genetic and protein biomarker of rhGH abuse in peripheral blood lymphocytes, serum, and plasma. However, logistic problems associated with current blood collection in sports drug testing point towards potential new alternative matrices that could be good candidates to be evaluated. Results obtained in this study showed high ELISA FN1 levels in one dried blood spot and in urine samples in ten healthy male volunteers treated with rhGH. Results showed that especially dried blood spots appear as a potential good matrix to detect rhGH abuse by means of FN1 biomarker. Copyright © 2016 John Wiley & Sons, Ltd.
Asunto(s)
Citocinas/sangre , Pruebas con Sangre Seca/métodos , Hormona de Crecimiento Humana/administración & dosificación , Detección de Abuso de Sustancias/métodos , Adulto , Doping en los Deportes , Ensayo de Inmunoadsorción Enzimática/métodos , Ejercicio Físico , Femenino , Fibronectinas , Hormona de Crecimiento Humana/sangre , Humanos , Masculino , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/sangre , Adulto JovenRESUMEN
Human Growth Hormone (hGH, somatotropin) is one of the relevant forbidden substances to be detected in sport drug testing. Since the appearance of recombinant hGH (rhGH) in the 80's, its expansion and availability through the black market have increased, so the detection of its abuse continues to be a challenge at present. New techniques or biomarkers that are robust, reliable, sensitive and allowing a large detection time window are welcome. rhGH produces an increase of insulin-like growth factor 1 (IGF-1). FN1 (fibronectin 1) and RAB31 (member of RAS oncogene family) genes have been suggested as two potential biomarkers for IGF-1 abuse. Following this line, in the present study some genetic and proteomic approaches have been performed with fourteen healthy male subjects treated with rhGH (which produces increase of IGF-1 concentrations) to study FN1 gene, FN1 protein, RAB31 gene and RAB31 protein as potential biomarkers for rhGH abuse. The results showed that both, RAB31 and FN1 genes and FN1 protein could be potential biomarkers for rhGH administration. Preliminary assessments of gender, age, acute sport activities and GHRP-2 (pralmorelin, a rhGH releasing peptide) influence suggest they are not relevant confounding factors. Thus, the selected markers present high sensitivity and a larger detection window for rhGH detection than IGF-1 itself.
Asunto(s)
Biomarcadores/sangre , Hormona de Crecimiento Humana/sangre , Adulto , Bioensayo/métodos , Doping en los Deportes/métodos , Femenino , Humanos , Factor I del Crecimiento Similar a la Insulina/metabolismo , Masculino , Proteómica/métodos , Proteínas Recombinantes/sangre , Deportes , Detección de Abuso de Sustancias/métodos , Adulto JovenRESUMEN
The everlasting pharmacological development is continuously producing new substances with potential doping abuse. Among these, secretagogues are very prone to misuse by athletes for their properties to release growth hormone (GH) and some limitations in the actual analytical methods to detect them. In this paper, an in-depth study on the key variables of the radio receptor method previously developed by our group is performed and a fit-for-purpose protocol is established. Thus, this sensitive and robust screening method is proposed as an intelligent and preventive antidoping method to detect new growth hormone secretagogues (GHSs) in exceptional suspicious urine samples obtained from athletes and will support the current detection methods based on liquid chromatography-mass spectrometry (LC-MS).
Asunto(s)
Doping en los Deportes , Glicoproteínas/orina , Hormona del Crecimiento/orina , Sustancias para Mejorar el Rendimiento/orina , Unión Competitiva , Cromatografía Liquida , Cromatografía de Gases y Espectrometría de Masas , Ghrelina/metabolismo , Células HEK293 , Humanos , Espectrometría de Masas , Receptores de Ghrelina/metabolismoRESUMEN
Immunohistochemistry (IHC) and in situ hybridization (ISH) techniques have been used for the detection of lymphocystis disease virus (LCDV) in formalin-fixed, paraffin-embedded tissues from gilt-head seabream, Sparus aurata L. Diseased and recovered fish from the same population were analysed. IHC was performed with a polyclonal antibody against a 60-kDa viral protein. A specific digoxigenin-labelled probe, obtained by PCR amplification of a 270-bp fragment of the gene coding the LCDV major capsid protein, was used for ISH. LCDV was detected in skin dermis and gill lamellae, as well as in several internal organs such as the intestine, liver, spleen and kidney using both techniques. Fibroblasts, hepatocytes and macrophages seem to be target cells for virus replication. The presence of lymphocystis cells in the dermis of the skin and caudal fin, and necrotic changes in the epithelium of proximal renal tubules were the only histological alterations observed in fish showing signs of the disease.
Asunto(s)
Infecciones por Virus ADN/veterinaria , Enfermedades de los Peces/patología , Enfermedades de los Peces/virología , Explotaciones Pesqueras/métodos , Hibridación in Situ/veterinaria , Iridoviridae/fisiología , Dorada/virología , Animales , Proteínas de la Cápside/inmunología , Infecciones por Virus ADN/patología , Infecciones por Virus ADN/virología , Genoma Viral , Inmunohistoquímica , Hibridación in Situ/métodosRESUMEN
Mx is an interferon-induced protein that protects against viral infections. In this study the absolute number of Mx transcripts after poly I:C injection (a synthetic dsRNA) or sole aquabirnavirus (solevirus) inoculation in Senegalese sole (Solea senegalensis Kaup) has been quantified. Mx expression profiles differed clearly in both experimental conditions; the induction response was faster and more intense after poly I:C injection than after solevirus inoculation. Moreover, pre-injection of soles with poly I:C prior to solevirus infection eliminated the induction of Mx expression associated with this virus. To evaluate the possible interference of poly I:C treatments on solevirus replication, the mRNA levels of the virus capsid protein (VP2) were determined by RT-PCR. VP2 transcripts were hardly detected in poly I:C pre-injected animals from 12 to 72 h after solevirus inoculation. All these data suggest that poly I:C is able to induce an antiviral state that interferes with solevirus replication, and support the suitability of Mx expression analysis as a marker to study the defensive response against solevirus.
Asunto(s)
Aquabirnavirus/inmunología , Enfermedades de los Peces/virología , Peces Planos/inmunología , Peces Planos/virología , Proteínas de Unión al GTP/genética , Poli I-C/inmunología , Transcripción Genética , Animales , Antivirales/inmunología , Infecciones por Birnaviridae/genética , Infecciones por Birnaviridae/veterinaria , Infecciones por Birnaviridae/virología , Proteínas de la Cápside/genética , Enfermedades de los Peces/genética , Peces Planos/genética , Proteínas de Resistencia a Mixovirus , Replicación Viral/efectos de los fármacosRESUMEN
Avaliou-se o efeito da dexmedetomidina sobre o ritmo cardíaco em 20 cães, sem raça definida, de ambos os sexos e considerados sadios, anestesiados pelo sevofluorano e submetidos a doses crescentes de adrenalina. Os animais foram, aleatoriamente, distribuídos em dois grupos (placebo e dexmedetomidina). No grupo placebo, os animais receberam, por via intravenosa, solução de NaCl a 0,9 por cento, na dose de 0,3ml/kg. Foram considerados dois momentos, M0 e M1, imediatamente antes e após a aplicação, respectivamente. Após 10 minutos, realizou-se a indução anestésica com sevofluorano, por meio de máscara facial vedada, até a perda do reflexo laringotraqueal. Em seguida, procedeu-se à intubação orotraqueal e a manutenção da anestesia foi realizada com a administração de sevofluorano na concentração de 1,5CAM, em circuito anestésico com reinalação parcial de gases. Decorridos 20 minutos da indução anestésica, iniciou-se a administração intravenosa contínua de solução de adrenalina a 2 por cento em doses crescentes de 1, 2, 3, 4 e 5mg/kg/min, por meio de bomba de infusão, com aumento da dose em intervalos de 10 minutos. Imediatamente antes desse acréscimo eram feitas as mensurações (M2 a M6). No grupo dexmedetomidina empregou-se a mesma metodologia substituindo-se a solução de NaCl a 0,9 por cento por hidrocloridrato de dexmedetomidina, na dose de 1µg/kg. Foram registradas as pressões arteriais, em M0 e em M2 a M6, e o traçado eletrocardiográfico, na derivação DII (M2 a M6), considerando-se para efeito estatístico o número total de bloqueios atrioventriculares (BAV) de primeiro e segundo graus e de complexos ventriculares prematuros (ESV), coincidentes com cada dose de adrenalina. Os dados foram submetidos à análise de variância seguida pelo teste de Tukey (P<0,05). Verificou-se que a dexmedetomidina interfere significativamente na condução atrioventricular levando a maior ocorrência de BAV e reduz o número de ESV nas doses infundidas de 2 e 3mg/kg/min ...
The effect of dexmedetomidine on the cardiac rhythm in twenty healthy mongrel dogs of both sexes anesthetized with sevofluorane and submitted to increasing doses of adrenaline was evaluated. The animals were randomly allotted to different treatment groups. Animals of placebo group were intravenous injected with 0.9 percent NaCl solution at 0.3ml/kg/IV. Two moments were considered, M0 and M1, the moments immediately before and after application, respectively. Ten minutes later, the dogs were anesthetized using sevofluorane via face mask until lost of their laringotracheal reflex. Then, orotracheal intubation was performed and maintenance of anesthesia was kept with 1,5MAC sevofluorane using an anesthetic circuit with a rebreathing system. Twenty minutes after anesthesia induction, continuous IV administration of 2 percent adrenaline solution was given at increasing doses of 1,2,3,4 and 5mg/kg/min., every ten minutes, respectively. The moments M2 to M6 were measured immediately before the next increase of dose. In dexmedetomidine group, the same technique was used replacing 0.9 percent NaCl by dexmedetomidine hydrochloride at 1mg/kg. Blood pressures were recorded at M0 and M2 to M6. Electrocardiography line in the derivation DII (M2 to M6) was used to observe the number of atrioventricular blocks (AVB) of first and second degrees and ventricular premature complexes (VPC). Statistic evaluation was performed by analysis of variance followed by Tukey's test (P<0.05). Dexmedetomidine significantly altered the atrioventricular conduction resulting in a higher occurrence of AVB. This drug reduced the number of VPC at 2 and 3mg/kg/min of adrenaline. After administration of dexmedetomidine, reduction of arterial blood pressure up to one hour and reduction of cardiac rate were observed
Asunto(s)
Animales , Perros , Anestesia por Inhalación/veterinaria , Dexmedetomidina/efectos adversos , Anestésicos/administración & dosificación , Anestésicos/efectos adversos , Arritmias Cardíacas/inducido químicamente , Arritmias Cardíacas/veterinariaRESUMEN
The role of tumour necrosis factor (TNF)-alpha or cyclo-oxygenase-2 (COX-2) eicosanoids in dystrophinopathies has been evaluated by chronically treating (4-8 weeks) adult dystrophic mdx mice with the anti-TNF-alpha etanercept (0.5 mg/kg) or the COX-2 inhibitor meloxicam (0.2 mg/kg). Throughout the treatment period the mdx mice underwent a protocol of exercise on treadmill in order to worsen the pathology progression; gastrocnemious muscles from exercised mdx mice showed an intense staining for TNF-alpha by immunohistochemistry. In vivo, etanercept, but not meloxicam, contrasted the exercise-induced forelimb force drop. Electrophysiological recordings ex vivo, showed that etanercept counteracted the decrease in chloride channel function (gCl), a functional index of myofibre damage, in both diaphragm and extensor digitorum longus (EDL) muscle, meloxicam being effective only in EDL muscle. None of the drugs ameliorated calcium homeostasis detected by electrophysiology and/or spectrofluorimetry. Etanercept, more than meloxicam, effectively reduced plasma creatine kinase (CK). Etanercept-treated muscles showed a reduction of connective tissue area and of pro-fibrotic cytokine TGF-beta1 vs. untreated ones; however, the histological profile was weakly ameliorated. In order to better evaluate the impact of etanercept treatment on histology, a 4-week treatment was performed on 2-week-old mdx mice, so to match the first spontaneous degeneration cycle. The histology profile of gastrocnemious was significantly improved with a reduction of degenerating area; however, CK levels were only slightly lower. The present results support a key role of TNF-alpha, but not of COX-2 products, in different phases of dystrophic progression. Anti-TNF-alpha drugs may be useful in combined therapies for Duchenne patients.
Asunto(s)
Ciclooxigenasa 2/metabolismo , Eicosanoides/metabolismo , Músculo Esquelético/metabolismo , Distrofia Muscular de Duchenne/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Creatina Quinasa/sangre , Creatina Quinasa/efectos de los fármacos , Inhibidores de la Ciclooxigenasa/farmacología , Electrofisiología , Etanercept , Inmunoglobulina G/farmacología , Inmunohistoquímica , Inmunosupresores/farmacología , Masculino , Meloxicam , Ratones , Ratones Endogámicos mdx , Microelectrodos , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patología , Condicionamiento Físico Animal , Receptores del Factor de Necrosis Tumoral , Tiazinas/farmacología , Tiazoles/farmacologíaRESUMEN
AIMS: The development and evaluation of a protocol based on polymerase chain reaction (PCR) and nucleic acid hybridization techniques for the specific detection of lymphocystis disease virus (LCDV) in several marine fish species. METHODS AND RESULTS: The pair of primers for PCR, OBL3 and OBL4, was designed based on published nucleotide sequence (LCDV-1) and amplifies a fragment within the major capsid protein. The sensitivity was evaluated using DNA from purified viral particles, as well as from cells inoculated with several viral concentrations. The PCR combined with slot blot was the most sensitive methodology, detecting 2.5 ng of viral DNA. Using this methodology LCDV was detected at 5 days postinoculation from SAF-1 cells initially inoculated with 10(-5) TCID(50) ml(-1). The combination of PCR with membrane hybridization has also been proved to be adequate to detect LCDV from apparently healthy carriers by means of caudal fin sample analysis. This asymptomatic infection was also demonstrated by classical virological methods (cell culture and immunoblot). CONCLUSIONS: The protocol described in this study allows the specific detection of LCDV, both in cell cultures and in fin homogenates from asymptomatic fish. SIGNIFICANCE AND IMPACT OF THE STUDY: The detection of asymptomatic carriers by a rapid molecular method using caudal fin sampling, which does not imply animal killing, could be an important tool to control epizootics caused by LCDV, as fish could be analysed before their introduction and/or mobilization in farm facilities.
Asunto(s)
Infecciones por Virus ADN/veterinaria , Enfermedades de los Peces/diagnóstico , Iridoviridae/aislamiento & purificación , Animales , Antígenos Virales/análisis , Secuencia de Bases , Línea Celular , Infecciones por Virus ADN/diagnóstico , ADN Viral/análisis , Peces/virología , Hibridación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Alineación de Secuencia/métodosRESUMEN
Human mammaglobin (hMAM) has recently been recognized as a breast associated glycoprotein. Although the biological role of hMAM is unknown, it has been previously reported that hMAM gene expression is a marker of low biological and clinical aggressiveness of breast cancer (BC). In this study, 148 cases of BC tissues were investigated for hMAM mRNA expression by reverse transcriptase-polymerase chain reaction (RT-PCR). In order to evaluate its prognostic value, hMAM was correlated with age of patients, type and size of tumor, nodal stage, histologic grade, c-erbB-2 over expression, Ki67 labelling index, estrogen receptor (ER) status and progesterone receptor (PGR) status. Fisher's exact test was used to examine the association between different parameters and hMAM. hMAM was expressed in 138/148 (93%) of BC tissues examined. Among the 10 hMAM negative cases, 8 were invasive ductal carcinomas (microscopically higher G3 grade) and 2 infiltrating lobular carcinomas. We found a significant association (p = 0.020) between absence of hMAM mRNA and G3 histologic grade but not with any other prognostic parameters studied. The present study indicates that lack of hMAM expression is restricted to the BC with G3 grading. Further studies are needed to clarify the biological basis and the clinical significance of our results.
Asunto(s)
Neoplasias de la Mama/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteínas de Neoplasias/biosíntesis , Uteroglobina/biosíntesis , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/patología , Línea Celular Tumoral , Femenino , Humanos , Antígeno Ki-67/biosíntesis , Mamoglobina A , Persona de Mediana Edad , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , ARN Mensajero/metabolismo , Receptores de Progesterona/metabolismo , Uteroglobina/genética , Uteroglobina/metabolismoRESUMEN
Occult carcinoma cells in peripheral blood of breast cancer (BC) patients is generally associated with poor disease prognosis. Reverse transcriptase-polymerase chain reaction (RT-PCR) is a sensitive method for revealing rare circulating cancer cells. The target mRNA must be carefully chosen, as it must be expressed only by malignant cells. In this study, we developed a nested RT-PCR assay for mammaglobin (hMAM) and assessed both its specificity and its sensitivity in the detection of cancer cells in peripheral blood of BC patients. hMAM mRNA was detected by RT-PCR in 156/165 (95%) of fresh BC tissues analyzed. All samples from 66 healthy blood donors and 151 patients with benign breast disease were hMAM negative as assessed by nested RT-PCR. In contrast, hMAM was detected in 16/137 (12%) of peripheral blood samples deriving from BC patients: 0/9 in stage 0, 1/50 (2%) in stage I, 3/33 (9%) in stage II, 1/18 (5%) in stage III and 11/27 (41%) in stage IV. Using nested RT-PCR, we were able to amplify hMAM transcript of one tumour cell/10(6) normal cells. Our data demonstrate that hMAM mRNA detection by RT-PCR is a specific assay potentially suitable for identification of occult cancer cells in peripheral blood of BC patients.
Asunto(s)
Biomarcadores de Tumor/sangre , Neoplasias de la Mama/sangre , Neoplasias de la Mama/diagnóstico , Proteínas de Neoplasias/biosíntesis , Células Neoplásicas Circulantes/patología , ARN Mensajero/metabolismo , Uteroglobina/biosíntesis , Línea Celular Tumoral , Humanos , Mamoglobina A , Sangre Oculta , Reacción en Cadena de la Polimerasa , Pronóstico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y EspecificidadRESUMEN
Antecedentes: El hiperparatiroidismo primario es causado por un adenoma único en alrededor del 90 por ciento de los casos; debido a la más precisa localización preoperatoria, se ha cuestionado la exploración cervical bilateral en todos los enfermos. Objetivos: Presentar la técnica y los resultados de la paratiroidectomia radioguiada mínimamente invasiva (PRM) en el tratamiento del hiperparatiroidismo primario. Lugar y aplicación: Servicio de cirugía oncológica. Diseño: Observacional retrospectivo. Población: 54 pacientes consecutivos operados por hiperparatiroidismo primario: 49 para tratamiento primario y 5 por recidiva. Método: Ecografía y centellograma con Sesta MIBI preoperatorios y paratiroidectomía radioguiada con cámara manual para detección de radiaciones gamma. Resultados: La cámara manual localizó al adenoma en el 88,8 por ciento de los casos, mientras que la ecografía y el centellograma lo hicieron en el 76 y 87 por ciento respectivamente. La combinación de los tres métodos permitió ubicar el adenoma en toda la serie; en 15 oportunidades fue necesario prolongar la cervicotomía por patología tiroidea sincrónica y en 2 por carcinoma de paratiroides. Conclusiones: La PRMI es una técnica complementaria de los estudios preoperatorios (AU)
Asunto(s)
Adulto , Masculino , Humanos , Femenino , Persona de Mediana Edad , Anciano , Hiperparatiroidismo/cirugía , Paratiroidectomía/métodos , Hiperparatiroidismo/diagnóstico por imagen , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Tecnecio Tc 99m Sestamibi/diagnóstico , Glándulas Paratiroides/cirugía , Glándulas Paratiroides/diagnóstico por imagen , Estudios RetrospectivosRESUMEN
Antecedentes: El hiperparatiroidismo primario es causado por un adenoma único en alrededor del 90 por ciento de los casos; debido a la más precisa localización preoperatoria, se ha cuestionado la exploración cervical bilateral en todos los enfermos. Objetivos: Presentar la técnica y los resultados de la paratiroidectomia radioguiada mínimamente invasiva (PRM) en el tratamiento del hiperparatiroidismo primario. Lugar y aplicación: Servicio de cirugía oncológica. Diseño: Observacional retrospectivo. Población: 54 pacientes consecutivos operados por hiperparatiroidismo primario: 49 para tratamiento primario y 5 por recidiva. Método: Ecografía y centellograma con Sesta MIBI preoperatorios y paratiroidectomía radioguiada con cámara manual para detección de radiaciones gamma. Resultados: La cámara manual localizó al adenoma en el 88,8 por ciento de los casos, mientras que la ecografía y el centellograma lo hicieron en el 76 y 87 por ciento respectivamente. La combinación de los tres métodos permitió ubicar el adenoma en toda la serie; en 15 oportunidades fue necesario prolongar la cervicotomía por patología tiroidea sincrónica y en 2 por carcinoma de paratiroides. Conclusiones: La PRMI es una técnica complementaria de los estudios preoperatorios