No effect of different bariatric surgery procedures on LINE-1 DNA methylation in diabetic and nondiabetic morbidly obese patients.

BACKGROUND: Bariatric surgery (BS) is proposed as a highly effective therapy for reducing weight and improving obesity-related co-morbidities. The molecular mechanisms involved in the metabolic improvement after BS are not completely resolved. Epigenetic modifications could have an important role. OBJECTIVE: The aim of this study was to evaluate the effect of different BS procedures (Roux-en-Y gastric bypass and laparoscopic sleeve gastrectomy) on global DNA methylation (long interspersed nucleotide element 1 [LINE-1]) in a group of nondiabetic and diabetic severely obese patients. SETTING: University hospital, Spain. METHODS: This study included 60 patients (30 nondiabetic and 30 diabetic severely obese patients) undergoing BS: 31 patients underwent Roux-en-Y gastric bypass and 29 underwent laparoscopic sleeve gastrectomy. Before and 6 months post-BS, anthropometric data, blood pressure, and metabolic parameters were determined. LINE-1 DNA methylation was quantified by pyrosequencing. We used the methylation levels of tumor necrosis factor-α as a control gene promoter. RESULTS: There were no differences between LINE-1 methylation levels at baseline and at 6 months after surgery (66.3±1.6 versus 66.2±2.06). Likewise, there was no statistically significant difference on LINE-1 methylation levels when we stratified according to metabolic status (diabetic versus nondiabetic), nor was there regarding the BS procedure. A strong correlation was shown between LINE-1 methylation levels and weight at baseline both in diabetic and nondiabetic obese patients (r = .486; P<.001). Tumor necrosis factor-α methylation levels increased significantly after BS in the group of diabetic obese patients. CONCLUSION: After BS, global LINE-1 methylation is not modified in the short term. More studies are required to determine if LINE-1 is a stable epigenetic marker, or, on the contrary, if it is susceptible to modification by external factors such as changes in lifestyle or a surgical intervention.

Mutation status and immunoglobulin gene rearrangements in patients from northwest and central region of Spain with chronic lymphocytic leukemia.

The aim of this study was to investigate the frequency and mutation status of the immunoglobulin heavy variable chain (IGHV) in a cohort of 224 patients from northwest and central region of Spain diagnosed with chronic lymphocytic leukemia (CLL), and to correlate it with cytogenetic abnormalities, overall survival (OS) and time to first treatment (TTFT). 125 patients had mutated IGHV, while 99 had unmutated IGHV. The most frequently used IGHV family was IGHV3, followed by IGHV1 and IGHV4. The regions IGHV3-30, IGHV1-69, IGHV3-23, and IGHV4-34 were the most commonly used. Only 3.1% of the patients belonged to the subfamily IGHV3-21 and we failed to demonstrate a worse clinical outcome in this subgroup. The IGHV4 family appeared more frequently with mutated pattern, similar to IGHV3-23 and IGHV3-74. By contrast, IGHV1-69 was expressed at a higher frequency in unmutated CLL patients. All the cases from IGHV3-11 and almost all from IGHV5-51 subfamily belonged to the group of unmutated CLL.

Identification of a novel recurrent gain on 20q13 in chronic lymphocytic leukemia by array CGH and gene expression profiling.

BACKGROUND: The presence of genetic changes is a hallmark of chronic lymphocytic leukemia (CLL). The most common cytogenetic abnormalities with independent prognostic significance in CLL are 13q14, ATM and TP53 deletions and trisomy 12. However, CLL displays a great genetic and biological heterogeneity. The aim of this study was to analyze the genomic imbalances in CLL cytogenetic subsets from both genomic and gene expression perspectives to identify new recurrent alterations. PATIENTS AND METHODS: The genomic imbalances and expression levels of 67 patients were analyzed. The novel recurrent abnormalities detected with bacterial artificial chromosome array were confirmed by FISH and oligonucleotide microarrays. In all cases, gene expression profiling was assessed. RESULTS: Copy number alterations were identified in 75% of cases. Overall, the results confirmed FISH studies for the regions frequently involved in CLL and also defined a new recurrent gain on chromosome 20q13.12, in 19% (13/67) of the CLL patients. Oligonucleotide expression correlated with the regions of loss or gain of genomic material, suggesting that the changes in gene expression are related to alterations in copy number. CONCLUSION: Our study demonstrates the presence of a recurrent gain in 20q13.12 associated with overexpression of the genes located in this region, in CLL cytogenetic subgroups.

Clonal plasma cells from monoclonal gammopathy of undetermined significance, multiple myeloma and plasma cell leukemia show different expression profiles of molecules involved in the interaction with the immunological bone marrow microenvironment.

The immunological bone marrow (BM) microenvironment plays a major role in controlling growth and survival of clonal plasma cells (PC); this might translate into different patterns of expression of molecules involved in immune responses on PC from different types of monoclonal gammopathies (MG). We have studied the expression of a group of nine such molecules on both BMPC and the plasma of 61 newly diagnosed MG patients (30 MG of undetermined significance (MGUS), 27 multiple myeloma (MM) and four plasma cell leukemia (PCL)) and five normal individuals. Clonal PC from all MG displayed significantly increased levels of CD56, CD86 and CD126, and decreased amounts of CD38 (P<0.001). Additionally, HLA-I and beta2-microglobulin were abnormally highly expressed in MGUS, while CD40 expression was decreased in MM and PCL (P<0.05). Interestingly, a progressive increase in the soluble levels of beta2-microglobulin was found from MGUS to MM and PCL patients (P=0.03). In contrast, all groups showed similar surface and soluble amounts of CD126, CD130 and CD95, except for increased soluble levels of CD95 observed in PCL. Overall, those phenotypic differences are consistent with increased antigen presentation and costimulatory capacities in MGUS, which progressively deteriorate in malignant MG (MM and PCL).

Carbamazepine induced transient monoclonal gammopathy and immunodeficiency.

Immune abnormalities have been found in many patients receiving anti-epileptic drugs. However, the effects of carbamazepine are still conflicting. We report the case of a 31-year-old woman who began carbamazepine treatment because of idiopathic epilepsy of adulthood. After three years of treatment she developed arthralgias and malaise. Complete immunologic evaluation showed a total absence of immunoglobulin M with decreased levels of immunoglobulin A, positive antinuclear antibodies and monoclonal paraproteinemia type IgG-kappa. The possibility of B cell lymphoma or myeloma was ruled out. Skin testing was negative. Bone marrow examination was normal. After carbamazepine discontinuation, levels of IgA and IgM increased until reaching normal values over 3 years. The monoclonal gammopathy of undetermined significance also disappeared over this period. During this period of immunodeficiency, the patient did not complain of any infectious complications.

Carbamazepine induced transient monoclonal gammopathy and immunodeficiency

Immune abnormalities have been found in many patients receiving anti-epileptic drugs. However, the effects of carbamazepine are still conflicting. We report the case of a 31-year-old woman who began carbamazepine treatment because of idiopathic epilepsy of adulthood. After three years of treatment she developed arthralgias and malaise. Complete immunologic evaluation showed a total absence of immunoglobulin M with decreased levels of immunoglobulin A, positive antinuclear antibodies and monoclonal paraproteinemia type IgG-kappa. The possibility of B cell lymphoma or myeloma was ruled out. Skin testing was negative. Bone marrow examination was normal. After carbamazepine discontinuation, levels of IgA and IgM increased until reaching normal values over 3 years. The monoclonal gammopathy of undetermined significance also disappeared over this period. During this period of immunodeficiency, the patient did not complain of any infectious complications (AU)

Se han detectado alteraciones inmunitarias en muchos pacientes que reciben antiepilépticos; sin embargo, los efectos de carbamacepina siguen siendo contradictorios. Describimos el caso de una mujer de 31 años que empezó a tomar carbamacepina por epilepsia idiopática en adultos. Después de tres años de tratamiento, experimentó artralgias y malestar. Una evaluación inmunológica completa reveló la ausencia total de inmunoglobulina M y disminución de la concentración de inmunoglobulina A, anticuerpos antinucleares positivos y una paraproteinemia monoclonal de tipo IgG-kappa. Se descartó la posibilidad de linfoma de los linfocitos B o mieloma. Las pruebas cutáneas fueron negativas. La exploración de la médula ósea fue normal. Después de suspender la carbamacepina, las concentraciones de IgA e IgM aumentaron hasta alcanzar sus valores normales a lo largo de 3 años. La gammapatía monoclonal de significación indeterminada también desapareció a lo largo de ese período. Durante ese período de inmunodeficiencia, la paciente no experimentó ninguna complicación infecciosa (AU)

Extended interval between enzyme therapy infusions for adult patients with Gaucher's disease type 1.

BACKGROUND: Enzyme replacement therapy (ERT) for Gaucher's disease with alglucerase or imiglucerase is efficacious, well-tolerated and safe. However, cost considerations, visits to medical facilities, potentially duration of theray for life, are issues of major concern to a proportion of treated patients and has, in some cases, led to the withdrawal of therapy. AIMS: To elucidate whether an extension of the interval between enzyme infusions to once every three weeks is as effective in maintaining the clinical responses achieved with the bi-monthly regimen. MATERIALS AND METHODS: Four patients with an optimal response to ERT (at 30 units/kg every two weeks for an average of 27 months), were subjected to enzyme dose/frequency changes that essentially constituted a reduction in cumulative dose over the treatment period. Patients were assessed every 6 months for alterations in haematological parameters, plasma chitotriosidase levels, liver and spleen size, and bone symptoms. RESULTS: All patients had to resume the previous infusion schedule of once every two weeks; one because of new bone marrow infiltrates, two because of visceral enlargement, and the fourth due to progressive anaemia. CONCLUSIONS: This limited experience suggests that a reduction in enzyme dose associated with an extended interval between infusions may lead to variable disease control, and underscores the need for individualization of enzyme therapy.

[Anemia secondary to lead poisoning. Our experience with 12 cases]. / Anemia secundaria a intoxicación por plomo. Nuestra experiencia a propósito de 12 casos.

BACKGROUND: A decrease in hemoglobin production is probably the main cause of anemia observed in patients poisoned by lead although hemolysis caused by the effects on membrane or by inhibition of other enzymes such as 5'-pyrimidine nucleotidase may also play a key role. The main lead exposure source comes from lead use in industry; however, outside occupational exposure, food and water consumption is the main cause of lead exposure. MATERIALS AND METHODS: Clinical and analytical characteristics are reported of twelve patients with lead poisoning of food and water source occurred in the last 14 years in two health areas in north Extremadura. RESULTS: The exposure source was wine, vinegar and olives in one case, hand-made brandy in five, water consumption in houses with lead piping, and in two cases the cause was not determined. The clinical picture was similar in all cases with the exception of one female patient who had encephalopathy and hepatic failure. Diagnosis was suggested by regenerative anemia with basophilic stippling of erythrocytes. All patients received oral calcium disodium EDTA. CONCLUSIONS: Lead poisoning in non-industrial areas is a rare entity. Nevertheless, owing to the use of artisanal procedures for wine elaboration and derivatives, its occurrence is increasing. In a patient with symptoms of lead poisoning and regenerative anemia, a peripheral blood smear should be reviewed.

Anemia secundaria a intoxicación por plomo. Nuestra experiencia a propósito de 12 casos / Anemia secondary to lead poisoning. Our twelve-case experience

Introducción. La disminución en la producción de hemo es probablemente la causa principal de la anemia observada en pacientes intoxicados por plomo, si bien la hemólisis provocada por los efectos sobre la membrana o por inhibición de otras enzimas tales como la 5'-pirimidin nucleotidasa también pueden tener una importancia capital. La mayor fuente de exposición al plomo proviene de su utilización en actividades industriales; sin embargo, fuera del ámbito laboral el origen alimentario es la principal causa de contacto con este tóxico. Material y métodos. Presentamos las características clínicas y analíticas de 12 casos de saturnismo de origen alimentario producidos en los últimos 14 años en 2 áreas sanitarias del norte de Extremadura. Resultados. La fuente de exposición fue vino, vinagre y aceitunas en un caso, aguardiente artesanal en 5, en 2 pacientes fue por el consumo de agua canalizada por tuberías de plomo y en 2 no se determinó la causa. El cuadro clínico fue superponible en todos, excepto en 1 paciente con encefalopatía y fallo hepático. La anemia regenerativa con punteado basófilo sugirió el diagnóstico. Todos recibieron tratamiento con ácido etilendiaminotetracético (EDTA) cálcico disódico oral. Conclusiones. La intoxicación por plomo en zonas no industriales constituye una rareza; sin embargo, debido al uso de procedimientos artesanales en la elaboración del vino y sus derivados nuevamente se está incrementando. En un paciente con síntomas de saturnismo y con anemia regrenerativa debe revisarse la extensión de sangre periférica (AU)

Agranulocitosis asociada a tratamiento con lamotrigina / Lamotrigine-induced agranulocytosis

No disponible

Hb Lepore-Baltimore (delta 68Leu-beta 84Thr) and Hb Lepore-Washington-Boston (delta 87Gln-beta IVS-II-8) in central Portugal and Spanish Alta Extremadura.

Hb Lepore is one of the most common abnormal haemoglobins in Caucasians in Central Portugal and in the Spanish Alta Extremadura (0.28% in a survey of school children). A group of 19 Portuguese and 14 Spanish Hb Lepore carriers (all unrelated) was characterised at the molecular level by the polymerase chain reaction, sequencing and restriction enzyme analysis. The Portuguese and one Spanish carrier were heterozygous for Hb Lepore-Baltimore, whereas all other Spanish subjects were Hb Lepore-Washington-Boston carriers. Sequencing of the Hb Lepore-Baltimore gene further established the crossover at delta 68-beta 84, a region two codons (CDs) shorter than that previously described and easily confirmed by digestion with MaeI and BanI. Data from haplotype analysis suggest that this crossover occurred as an independent event on the Iberian Peninsula. The haematological data were similar in both groups except for the levels of Hb F and the G gamma chain, which were significantly higher in the Hb Lepore-Baltimore heterozygotes. Quantification of the globin chains and the mRNA transcripts showed that the delta beta gene is transcribed at a higher level than the delta gene with levels of translation giving rise to 10%-15% of Hb Lepore. The different levels of Hb F observed in the two groups are the results of the higher transcription rate of the gamma genes in Hb Lepore-Baltimore heterozygotes and an apparently less efficient translation of G gamma genes in Hb Lepore-Washington-Boston heterozygotes.

[Molecular analysis of glucose-6-dehydrogenase deficiency in Spain]. / Análisis molecular del déficit de glucosa-6-fosfato deshidrogenasa (G6PD) en España.

PURPOSE: G6PD deficiency is the most frequent enzymopathy-producing genetic polymorphism in humans. Up to now, over 400 putative variants of G6PD have been distinguished on the basis of biochemical characterization of the deficient enzyme. Analysis of the G6PD gene has made possible a precise classification of the G6PD molecular variants by identification of about 80 different point mutations causing much of the phenotypic heterogeneity. In the Spanish population, the analysis of G6PD has led to the identification of 15 different point mutations that underlay the phenotypic heterogeneity of G6PD previously reported by biochemical analysis. The purpose of the study has been to identify the genetic mutation responsible of the G6PD deficiency and to improve the knowledge of its genetic homogeneity. PATIENTS AND METHODS: From 50 Spanish males with G6PD deficiency 34 came from out consultation and 16 from the Spanish Study Group on Red Cell Pathology (GEHBTA-Eritropatología) The methods employed included screening of prevalent mutations by ER-PCR, SSCP-PCR, genetic segmentation and biochemical characterization of the deficient enzyme. RESULTS: In 31 cases the mutations were characteristic of the four most frequent polymorphic variants found in Spain (G6PD A-376G/202A, G6PD Mediterranean 563T G6PD Union 1360T and G6PD Seattle 344C). Since these mutations either create or abolish a specific site recognized by a restriction endonuclease (RE), they can be rapidly detected by RE digestion of a PCR-amplified product (PCR-RE). In patients where none of these mutations were present (17 cases), the G6PD gene was subjected to PCR single-strand conformation polymorphism (PCR-SSCP) analysis combined with direct PCR-sequencing. By using this procedure, 9 new mutations have been identified, five of them have been also found in other geographical areas and were associated with favism (G6PD A-376G/968C, G6PD Santamaria 376G/542T, G6PD Aures 143C and G6PD Chatham 1003A) or chronic haemolytic anaemia (G6PD Tomah 1153C). The other four mutations are unique and not reported so far: Three of them are associated with favism (G6PD Málaga 542T, G6PD Murcia 209G and G6PD Valladolid 406T) and one with chronic haemolytic anaemia (G6PD Madrid 1155G). The remaining eight cases are under study. CONCLUSION: The present study confirms the marked genetic heterogeneity of G6PD deficiency in Spain and demonstrate that the PCR-RE analysis is an easy tool for rapid diagnosis of the molecular defect in subjects with the common forms of G6PD deficiency. Furthermore the fact that G6PD A-376G/202A is the most common variant within Spanish population and the finding of G6PD Aures 43C and G6PD Santamaría 76G/542T, who are polymorphic in Algeria is consistent with a significant gene flow from Africa to Europe through Spain.

[Extreme increase in the blood erythroblast count in a patient with iron-deficiency anemia and asplenia]. / Eritroblastemia intensa en un paciente con anemia ferropénica y asplenia.

Bone marrow stress can provoke the presence of high erythroblast count in peripheral blood of patients with splenectomy, but is very rare in patients with normal spleen. Hypoxia occurring in patients with severe ferropenic anaemia could originate the occasional presence of erythroblasts in peripheral blood. An associated pathology should be excluded if there is a high normoblast count. We present the case of a 40 years old woman with ferropenic anaemia and a very high normoblast count. Hyperplastic polips were found in the gastric tissue as the cause for her anaemia. The absence of spleen explained the presence of erytroblasts. Further studies excluded the possibility of hemoglobinopathy and bone marrow studies were normal. Treatment with iron was successfully in correcting the hemoglobin levels and the normoblasts disappeared from peripheral blood also. A careful examination of the peripheral blood film and, specially the presence of Howell-Jolly bodies is the cornerstone of the possible diagnosis confirmed with the appropriate radiological studies.