Enfermedad de Pompe de inicio tardío: análisis de una casuística de 19 pacientes mexicanos / Late onset Pompe disease: an analysis of 19 patients from Mexico

Introducción: La enfermedad de Pompe es una miopatía metabólica rara con espectro clínico heterogéneo, especialmente la de inicio tardío, cuya sintomatología es de progresión más lenta y representa un gran reto diagnóstico. Objetivo: Describir el genotipo y las características clínicas de pacientes mexicanos con Pompe de inicio tardío (LOPD). Material y métodos. Se incluyó a 19 pacientes mexicanos con LOPD confirmada mediante actividad enzimática y estudio molecular del gen GAA. Se evaluaron datos clínicos y se revisaron las mutaciones en bases de datos genómicas. Resultados: La mediana de edad de inicio de los síntomas fue de 19 años (rango: 2-43 años), y la edad de diagnóstico, de 36 años (rango: 9-52 años). Los síntomas más frecuentes fueron debilidad axial y proximal (n = 17; 89,5%), marcha basculante (n = 17; 89,5%) e hiperlordosis (n = 7; 36,8%). A 16 pacientes (84,2%) se les realizó electromiografía; 11 (57,8%) describieron patrón miopático y sólo en cinco pacientes (26%) se incluyó la valoración de los músculos paraespinales. Las variantes patogénicas más frecuentes en nuestra casuística fueron c.-32-13T>G, c.1799G>A y c.1082C>T. Conclusiones: Parecido a lo comunicado en publicaciones internacionales, la LOPD en México es clínicamente heterogénea; los pacientes pueden tardar años en llegar al diagnóstico. La debilidad muscular axial y proximal es el dato clínico más frecuente, por lo que la electromiografía debe incluir valoración de los músculos paraespinales. A excepción de una, las mutaciones encontradas en nuestra serie de casos se encuentran previamente descritas en las bases de datos de enfermedad de Pompe.(AU)

Introduction: Pompe disease (PD) is a rare metabolic myopathy with an ample and heterogeneous clinical spectrum, particularly late onset PD (LOPD), which is characterized by appearance at older age and slower disease progression, leading to diagnostic confirmation difficulty and delay. Aim: To describe the genotype and clinical characteristics of Mexican patients with LOPD. Material and methods: Clinical information from 19 Mexican patients with LOPD confirmed with enzyme activity and GAA gene analysis was reviewed. Genetic information of our population was crossed with international genetic databases. Results: Median age between onset of symptoms and diagnosis was 19 years (range 2-43) and diagnostic confirmation 36 years (range 9-52). Most frequently referred symptoms were proximal axial weakness (n = 17; 89.5%), waddling gait (n = 17; 89.5%) and hyperlordosis (n = 7; 36.8%). Sixteen patients (84.2%) were evaluated with electromyography; a myopathic pattern was reported in 11 (57.8%), but only in 5 patients (26%) paraspinal muscle evaluation was included. The most pathogenic mutations in our group were c.-32-13T>G, c.1799G>A and c.1082C>T. Conclusions: Similar to other international publications, LOPD in Mexico is clinically heterogeneous; patients may delay years before diagnosis is established. Axial and proximal weakness is the most frequent clinical feature; thus, electromyography with paraspinal muscle evaluation is essential. Except for one, the mutations found in our patients have been previously reported in PD genetic databases.(AU)

[Late onset Pompe disease: an analysis of 19 patients from Mexico]. / Enfermedad de Pompe de inicio tardío: análisis de una casuística de 19 pacientes mexicanos.

INTRODUCTION: Pompe disease (PD) is a rare metabolic myopathy with an ample and heterogeneous clinical spectrum, particularly late onset PD (LOPD), which is characterized by appearance at older age and slower disease progression, leading to diagnostic confirmation difficulty and delay. AIM: To describe the genotype and clinical characteristics of Mexican patients with LOPD. MATERIAL AND METHODS: Clinical information from 19 Mexican patients with LOPD confirmed with enzyme activity and GAA gene analysis was reviewed. Genetic information of our population was crossed with international genetic databases. RESULTS: Median age between onset of symptoms and diagnosis was 19 years (range 2-43) and diagnostic confirmation 36 years (range 9-52). Most frequently referred symptoms were proximal axial weakness (n = 17; 89.5%), waddling gait (n = 17; 89.5%) and hyperlordosis (n = 7; 36.8%). Sixteen patients (84.2%) were evaluated with electromyography; a myopathic pattern was reported in 11 (57.8%), but only in 5 patients (26%) paraspinal muscle evaluation was included. The most pathogenic mutations in our group were c.-32-13T>G, c.1799G>A and c.1082C>T. CONCLUSIONS: Similar to other international publications, LOPD in Mexico is clinically heterogeneous; patients may delay years before diagnosis is established. Axial and proximal weakness is the most frequent clinical feature; thus, electromyography with paraspinal muscle evaluation is essential. Except for one, the mutations found in our patients have been previously reported in PD genetic databases.

TITLE: Enfermedad de Pompe de inicio tardío: análisis de una casuística de 19 pacientes mexicanos.Introducción. La enfermedad de Pompe es una miopatía metabólica rara con espectro clínico heterogéneo, especialmente la de inicio tardío, cuya sintomatología es de progresión más lenta y representa un gran reto diagnóstico. Objetivo. Describir el genotipo y las características clínicas de pacientes mexicanos con Pompe de inicio tardío (LOPD). Material y métodos. Se incluyó a 19 pacientes mexicanos con LOPD confirmada mediante actividad enzimática y estudio molecular del gen GAA. Se evaluaron datos clínicos y se revisaron las mutaciones en bases de datos genómicas. Resultados. La mediana de edad de inicio de los síntomas fue de 19 años (rango: 2-43 años), y la edad de diagnóstico, de 36 años (rango: 9-52 años). Los síntomas más frecuentes fueron debilidad axial y proximal (n = 17; 89,5%), marcha basculante (n = 17; 89,5%) e hiperlordosis (n = 7; 36,8%). A 16 pacientes (84,2%) se les realizó electromiografía; 11 (57,8%) describieron patrón miopático y sólo en cinco pacientes (26%) se incluyó la valoración de los músculos paraespinales. Las variantes patogénicas más frecuentes en nuestra casuística fueron c.-32-13T>G, c.1799G>A y c.1082C>T. Conclusiones. Parecido a lo comunicado en publicaciones internacionales, la LOPD en México es clínicamente heterogénea; los pacientes pueden tardar años en llegar al diagnóstico. La debilidad muscular axial y proximal es el dato clínico más frecuente, por lo que la electromiografía debe incluir valoración de los músculos paraespinales. A excepción de una, las mutaciones encontradas en nuestra serie de casos se encuentran previamente descritas en las bases de datos de enfermedad de Pompe.

Transcriptional activity and splicing factors are preserved during physiological apoptosis.

Apoptosis is the best-known programmed cell death that maintains tissue homeostasis in eukaryotic cells. The morphological characteristics include nuclear and cytoplasmic contraction and cytoplasmic blebbing, its biochemical hallmarks include caspase protease activity and DNA fragmentation. In rat ovaries, cell death is a normal process that occurs throughout the organism's life. Granulosa cells, the more abundant cell type forming the ovarian follicles, are eliminated via different routes of cell death. Most granulosa cells are eliminated through apoptotic cell death. In this work, we analyzed the behavior of nuclear components throughout the apoptotic process and determined how they are regionalized and conserved during follicular atresia in rat ovaries. Apoptosis was detected based on caspase-3 activity and DNA fragmentation using the TUNEL technique. We identified the transcription markers H3ac and RNA Pol II, and splicing factor SC35 by immunodetection. The nucleolar components were analyzed via light microscopy and transmission electron microscopy through immunodetection of the proteins nucleolin and nucleophosmin-1. The nuclear ultrastructure was analyzed using standard contrast and preferential ribonucleoprotein contrast. Our results demonstrate that during the progression of apoptosis, chromatin is remodeled to constitute apoptotic bodies; transcription and spliceosome elements are reorganized along with the nucleolar components. Additionally, the splicing and transcription factors are segregated into specific territories inside the apoptotic bodies, suggesting that transcriptional elements are reorganized during the apoptotic process. Our results indicate that apoptotic bodies not only are compacted, and chromatin degraded but all the nuclear components are progressively reorganized during cell elimination; moreover, the transcriptional components are preserved.

Apoptotic and autophagic cell death induced by glucolaxogenin in cervical cancer cells.

The antiproliferative and cytotoxic activity of glucolaxogenin and its ability to induce apoptosis and autophagy in cervical cancer cells are reported. We ascertained that glucolaxogenin exerts an inhibitory effect on the proliferation of HeLa, CaSki and ViBo cells in a dose-dependent manner. Analysis of DNA distribution in the cell-cycle phase of tumor cells treated with glucolaxogenin suggests that the anti-proliferative activity of this steroid is not always dependent on the cell cycle. Cytotoxic activity was evaluated by detection of the lactate dehydrogenase enzyme in supernatants from tumor cell cultures treated with the steroid. Glucolaxogenin exhibited null cytotoxic activity. With respect to the apoptotic activity, the generation of apoptotic bodies, the presence of active caspase-3 and annexin-V, as well as the DNA fragmentation observed in all tumor lines after treatment with glucolaxogenin suggests that this compound does indeed induce cell death by apoptosis. Also, a significantly increased presence of the LC3-II, LC3 and Lamp-1 proteins was evidenced with the ultrastructural existence of autophagic vacuoles in cells treated with this steroidal glycoside, indicating that glucolaxogenin also induces autophagic cell death. It is important to note that this compound showed no cytotoxic effect and did not affect the proliferative capacity of mononuclear cells obtained from normal human peripheral blood activated by phytohaemagglutinin. Thus, glucolaxogenin is a compound with anti-proliferative properties that induces programmed cell death in cancer cell lines, though it is selective with respect to normal lymphocytic cells. These findings indicate that this glycoside could have a selective action on tumor cells and, therefore, be worthy of consideration as a therapeutic candidate with anti-tumor potential.

Analysis of different cell death processes of prepubertal rat oocytes in vitro.

The processes of cell death were studied in vitro in populations of oocytes isolated from prepubertal rats. In order to identify apoptosis, the externalized phosphatidylserine was recognized with Annexin-V coupled to FITC and the fragmentation of DNA was demonstrated by means of electrophoresis. Oocytes were tested for autophagy by means of the incorporation of monodansylcadaverine and monitoring Lc3-I/Lc3-II by western blot. The expression of mRNA marker genes of autophagy and of apoptosis was studied by means of RT-PCR in pure populations of oocytes. Some oocytes expressed at least one of the following markers: caspase-3, lamp1 and Lc3. Some oocytes were positive to Annexin-V or to monodansylcadaverine. However, most of them were simultaneously positive to both markers. The relative frequency of oocytes simultaneously positive to markers of apoptosis and autophagy did not change in the different ages studied. The transformation of Lc3-I in Lc3-II was present in all populations of oocytes studied. The mRNAs for caspase-3, lamp1 and Lc3 were present in all populations of oocytes analyzed. Our results demonstrate that oocytes of rats from new born to prepubertal age are eliminated by means of three different cell death processes: apoptosis, autophagy and a mixed event in which both routes to cell death participate in the same cell.

Diagnósticos de enfermería prevalentes en cuidados paliativos oncológicos / No disponible

Objetivo: determinar cuáles son los diagnósticos de enfermería más prevalentes en cuidados paliativos oncológicos. Verificar que los diagnósticos consensuados previamente en nuestro grupo de trabajo se encuentran entre los más prevalentes. Material y métodos: estudio descriptivo multicéntrico y retrospectivo de prevalencia. La muestra la forman 1.255 pacientes oncológicos de edad superiora 18 años, incluidos en programas de 7 unidades de cuidados paliativos localizadas en La Coruña, Málaga, Algeciras, Madrid, Barcelona, Salamanca y Santander. Fueron excluidos los pacientes no oncológicos y los no incluidos en los programas anteriormente mencionados. Los datos fueron recogidos desde el 1 de enero de 2007 al 31 de diciembre de 2007. Se realizó una revisión de las historias clínicas identificando los diagnósticos de enfermería registrados durante todo el proceso asistencial. Posteriormente se calculó la prevalencia con los datos obtenidos. Se emplearon medidas de nivel descriptivo, tanto absolutas como relativas. Resultados: se revisaron 1.255 historias obteniéndose un total de 6.351 diagnósticos. Los más prevalentes fueron: dolor crónico 67,4% (876), intolerancia a la actividad 66,9% (871), riesgo de estreñimiento 61,8% (803), deterioro del patrón del sueño 55,7% (724), deterioro de la mucosa oral 53,1% (691), riesgo de deterioro de la integridad cutánea 41,1% (529), ansiedad 35,3% (459), riesgo de cansancio de desempeño del rol de cuidador 35,0% (455), baja autoestima situacional 28,0% (372), sufrimiento espiritual 24,5% (319) y afrontamiento familiar comprometido 19,3% (252). Conclusiones: los diagnósticos más prevalentes coinciden con los consensuados previamente en nuestro grupo de trabajo. A la luz de estos resultados el presente estudio da paso al diseño de planes de cuidados estandarizados para pacientes paliativos oncológicos basándose en estos diagnósticos (AU)

Objective: to identify the most prevalent nursing diagnoses in oncological palliative-care patients. To verify that the nursing diagnoses previously considered most prevalent by our work group were among them. Material and methods: a descriptive, multicenter, retrospective study of prevalence. The sample included 1,255 oncological patients older than 18 years of age who were included in a program involving 7 palliative care units in La Coruña, Málaga, Algeciras, Madrid, Barcelona, Salamanca, and Santander. Non-cancer patients were excluded, as were patients not included in the above-mentioned program. Data were collected from January 1, 2007 to December 31, 2007. A review of the medical records with a nursing diagnosis was performed. Subsequently, prevalence was calculated. Absolute and relative measurements were used in the descriptive analysis. Results: 1,255 medical records were reviewed for a total of 6,351 diagnoses. Most prevalent diagnoses were: chronic pain 67.4% (876), activity intolerance 66.9% (871), risk of constipation 61.8% (803), sleep pattern disturbance 55.7% (724), oral mucosal impairment 53.1% (691), risk of skin integrity impairment 41.1% (529), anxiety 35.3% (459), risk of caregiver stress 35.0% (455), situational self-esteem disturbance 28.0% (372), spiritual distress 24.5% (319) and family coping impairment 19.3% (252). Conclusions: the most prevalent nursing diagnoses found match up with the diagnoses previously agreed upon by our work group. In the light of these results the present study makes way for the design of standard nursing care plans for oncological palliative-care patients based on these diagnoses (AU)

Hematologic abnormalities in liver-transplanted children during medium- to long-term follow-up.

OBJECTIVE: Hematologic abnormalities as adverse effects related to immunosuppressive drugs in liver-transplanted children are rarely reported. We have observed anemia, neutropenia, and thrombocytopenia in our pediatric liver-transplant population. The aim of this study was to exclude all suspected etiologies to define the association of immunosuppressants with these abnormalities. METHODS: Patients under 18 years old who still attend periodic controls at liver-transplant outpatient clinics were considered. Seventy patients met the inclusion criteria, 36 girls and 34 boys. Mean patient age was 5.6 years (range: 7 months to 17 years) and mean follow-up 6 years (range: 1-10 years). Medical records were reviewed beginning 1 month posttransplant. Treatment exposures, irradiation, blood product administration, and all laboratory studies were reviewed. When a hematologic abnormality was detected, we recorded the management for its resolution, the clinical response to therapy and the length of treatment. RESULTS: Twenty-five of the 70 children suffered 26 abnormal hematologic episodes (anemia 14, neutropenia 2, thrombocytopenia 3, simultaneous anemia and neutropenia 5, and pancytopenia 2). Eleven episodes (42%) had unclear etiologies and the process of elimination suggested an association with the immunosuppressant. Switching immunosuppressant was required in four patients and dose reduction in seven. CONCLUSIONS: Hematologic abnormalities in liver-transplanted children are common. The etiology is readily attributable to several causes. When the immunosuppressant appears to be a possible cause, the first step is dose reduction. If the hematologic abnormality persists despite dose reduction, a trial switch may be required.

[First orthotopic liver transplantation in patient with biliary atresia and situs inversus in spain]. / Primer trasplante hepático en un paciente con atresia de vías biliares y situs inversus en España.

Biliary atresia is the most common indication for liver transplantation in the pediatric age group. The Kasai portoenterostomy has become established as the primary treatment for biliary atresia. If portoenterostomy fails, death before 2 years of age is likely without liver transplantation. The most common multiple malformation syndrome associated with biliary atresia is polysplenia syndrome, which forms a constellation of defects of body symmetry, splenic development and vascular anomalies, including situs inversus, polysplenia and others. The situs inversus was formerly considered an absolute contraindication for liver transplantation. Recently however, several case reports have been published suggesting that neither situs inversus nor this particular subset of vascular abnormalities should be considered contraindications to liver transplantation. We present one case of liver transplantation performed in patient with biliary atresia, situs inversus and polysplenia. This is the first report described in Spain for a liver transplant in a child with biliary atresia plus situs inversus.

Primer trasplante hepático en un paciente con atresia de vías biliares y situs inversus en España / First orthotopic liver transplantation in patient with biliary atresia and situs inversus in Spain

La atresia de vías biliares es la indicación más frecuente para trasplante hepático en la edad pediátrica. La portoenterostomía de Kasai es el tratamiento primario para la atresia de vías biliares. Si la portoenterostomía falla, el fallecimiento suele ocurrir antes de los 2años de edad sin el trasplante hepático. El síndrome de múltiples malformaciones más comunmente asociado con la atresia de vías biliares es el síndrome de poliesplenia, formado por una constelación de defectos en la simetría corporal, desarrollo del bazo y anomalías vasculares, incluyendo situs inversus, poliesplenia y otros. El situs inversus era considerado como una contraindicación absoluta para el trasplante hepático. Sin embargo recientemente han sido publicados varios casos, sugiriendo que ni el situs inversus ni la presencia de dichas alteraciones vasculares deben ser consideradas como contraindicaciones para el trasplante hepático. Presentamos un caso de trasplante hepático realizado a un paciente con atresia de vías biliares, situs inversus y poliesplenia. Es el primer caso descrito en España (AU)

Isolation of a proliferation inhibitor factor from uterine myomatosis fibroblasts.

In this work, we report the isolation of a factor from the culture supernatant of confluent fibroblasts from human cervix with the diagnosis of uterine myomatosis. This factor possesses the capacity to inhibit the proliferation of normal fibroblasts. The proliferation inhibitor factor (PIF) was purified from the culture supernatant by precipitation with 80% ammonium sulfate, and by molecular sieve chromatography. Our results indicate that PIF is a protein of 23 kDa, which is highly sensitive to trypsin treatment, and is thermolabile, since temperatures equal to, or above, 60 degrees C eliminate the protein activity in 15 to 20 min. Western blot analyses identified no cross reactions of the purified PIF with TGF-alpha, TNFalpha, IFNgamma, or IL-1beta, suggesting that PIF is a new protein belonging to the group of factors secreted by fibroblasts able to inhibit cellular proliferation.

[Hematopoietic growth factors induce proliferation of non-hematopoietic tumor cells]. / Los factores de crecimiento hematopoyéticos estimulan la proliferación de células tumorales no hematopoyéticas.

OBJECTIVE: To evaluate the effect of hematopoietic growth factors (HGF) on the proliferation of non-hematopoietic cells such as fibroblasts and epithelial cells of normal and tumoral origin. METHODS: The lymphoid factor IL-2 and the myeloid HGF assayed were IL-3, G-CSF, GM-CSF and M-CSF. The cellular proliferation was determined by measuring the amount of crystal violet dye incorporation by cells through spectrophotometry. RESULTS: All myeloid HGF tested stimulated the proliferation of cell lines 5637, CaLo and L-929. These results suggest that the stromal cells can be induced to proliferate by myeloid growth factors hinting to a bilateral interaction between these two types of cells as it is known that stromal cells in turn secrete HGF. We also observed that for the mouse fibroblastic and epithelial cells, IL-2 was unable to induce proliferation in normal cells but had a strong effect on transformed cells. Finally we discuss our observation that tumour cells responded to IL-2 as a possible mechanism for an immune escape by these cells through IL-2 depletion.