Pathological investigation of high pathogenicity avian influenza H5N8 in captive houbara bustards (Chlamydotis undulata), the United Arab Emirates 2020.

At the end of 2020, an outbreak of HPAI H5N8 was registered in captive African houbara bustards (Chlamydotis undulata) in the United Arab Emirates. In order to better understand the pathobiology of this viral infection in bustards, a comprehensive pathological characterization was performed. A total of six birds were selected for necropsy, histopathology, immunohistochemistry, RNAscope in situ hybridization and RT-qPCR and nanopore sequencing on formalin-fixed and paraffin-embedded (FFPE) tissue blocks. Gross lesions included mottled and/or hemorrhagic pancreas, spleen and liver and fibrinous deposits on air sacs and intestine. Necrotizing pancreatitis, splenitis and concurrent vasculitis, hepatitis and fibrino-heterophilic peritonitis were identified, microscopically. Viral antigens (nucleoprotein) and RNAs (matrix gene) were both detected within necro-inflammatory foci, parenchymal cells, stromal cells and endothelial cells of affected organs, including the myenteric plexus. Molecular analysis of FFPE blocks successfully detected HPAI H5N8, further confirming its involvement in the lesions observed. In conclusion, HPAI H5N8 in African houbara bustards results in hyperacute/acute forms exhibiting marked pantropism, endotheliotropism and neurotropism. In addition, our findings support the use of FFPE tissues for molecular studies of poorly characterized pathogens in exotic and endangered species, when availability of samples is limited.

The effects of an 8-week hypopressive exercise training program on urinary incontinence and pelvic floor muscle activation: A randomized controlled trial.

BACKGROUND: Pelvic floor dysfunction and urinary incontinence are two of the most frequent gynecological problems, and pelvic floor muscle training is recommended as a first-line treatment, with new approaches such as hypopressive exercises. This study aimed to analyze the efficacy of an 8-week supervised training program of hypopressive exercises on pelvic floor muscle strength and urinary incontinence symptomatology. DESIGN: Blinded randomized controlled trial. SETTINGS: Women with pelvic floor dysfunction and urinary incontinence symptoms, aged 18-60 years. PARTICIPANTS: A total of 117 participants were randomly allocated to the hypopressive exercises group (n = 62) or a control group that received no intervention (n = 55) and completed the study. MAIN OUTCOME MEASURES: Clinical and sociodemographic data were collected, as well as pelvic floor muscle strength (using the Modified Oxford Scale); the genital prolapse symptoms, colorectal symptoms, and urinary symptoms (with the Pelvic Floor Distress Inventory [PFDI-20]); the impact of pelvic floor disorders (PFD) on women's lives (with the Pelvic Floor Impact Questionnaire [PFIQ-7]); and the severity of urinary incontinence symptoms (using the International Consultation on Incontinence Questionnaire [ICIQ]). RESULTS: The results showed an improvement in the hypopressive group in the pelvic floor muscle strength F (1117) = 89.514, p < 0.001, a significantly lower score for the PFIQ7 total score, t (112) = 28.895, p < 0.001 and FPDI20 t (112) = 7.037, p < 0.001 as well as an improvement in ICIQ-SF values after 8 weeks of intervention in comparison with the control group. CONCLUSIONS: After performing an 8-week of hipopressive exercises intervention, a decrease in pelvic floor disorders associated symptoms can be observed. In addition, pelvic floor muscle contractility is improved and a decrease in severity and symptoms associated with urinary incontinence has been reported.

HDAC11 is a novel regulator of fatty acid oxidative metabolism in skeletal muscle.

Skeletal muscle is the largest tissue in mammalian organisms and is a key determinant of basal metabolic rate and whole-body energy metabolism. Histone deacetylase 11 (HDAC11) is the only member of the class IV subfamily of HDACs, and it is highly expressed in skeletal muscle, but its role in skeletal muscle physiology has never been investigated. Here, we describe for the first time the consequences of HDAC11 genetic deficiency in skeletal muscle, which results in the improvement of muscle function enhancing fatigue resistance and muscle strength. Loss of HDAC11 had no obvious impact on skeletal muscle structure but increased the number of oxidative myofibers by promoting a glycolytic-to-oxidative muscle fiber switch. Unexpectedly, HDAC11 was localized in muscle mitochondria and its deficiency enhanced mitochondrial content. In particular, we showed that HDAC11 depletion increased mitochondrial fatty acid ß-oxidation through activating the AMP-activated protein kinase-acetyl-CoA carboxylase pathway and reducing acylcarnitine levels in vivo, thus providing a mechanistic explanation for the improved muscle strength and fatigue resistance. Overall, our data reveal a unique role of HDAC11 in the maintenance of muscle fiber-type balance and the mitochondrial lipid oxidation. These findings shed light on the mechanisms governing muscle metabolism and may have implications for chronic muscle metabolic disease management.

Loss of HDAC11 accelerates skeletal muscle regeneration in mice.

Histone deacetylase 11 (HDAC11) is the latest identified member of the histone deacetylase family of enzymes. It is highly expressed in brain, heart, testis, kidney, and skeletal muscle, although its role in these tissues is poorly understood. Here, we investigate for the first time the consequences of HDAC11 genetic impairment on skeletal muscle regeneration, a process principally dependent on its resident stem cells (satellite cells) in coordination with infiltrating immune cells and stromal cells. Our results show that HDAC11 is dispensable for adult muscle growth and establishment of the satellite cell population, while HDAC11 deficiency advances the regeneration process in response to muscle injury. This effect is not caused by differences in satellite cell activation or proliferation upon injury, but rather by an enhanced capacity of satellite cells to differentiate at early regeneration stages in the absence of HDAC11. Infiltrating HDAC11-deficient macrophages could also contribute to this accelerated muscle regenerative process by prematurely producing high levels of IL-10, a cytokine known to promote myoblast differentiation. Altogether, our results show that HDAC11 depletion advances skeletal muscle regeneration and this finding may have potential implications for designing new strategies for muscle pathologies coursing with chronic damage. DATABASE: Data were deposited in NCBI's Gene Expression Omnibus accessible through GEO Series accession number GSE147423.

Exposure to and Circulation of Avian Influenza and Newcastle Disease Viruses in Peridomestic Wild Birds in the United Arab Emirates.

Avian influenza and Newcastle disease viruses (AIV, NDV) are major pathogens of captive and wild birds worldwide. Wetlands and their associated bird communities, especially waterfowl and shorebirds, are known to play a central role in the epidemiology of these diseases as maintenance hosts. However, these viruses also circulate in places where these ecosystems and communities are uncommon, suggesting the involvement of other taxa in their epidemiological cycles. In the arid region of Abu Dhabi of the United Arab Emirates (UAE), both viruses are regularly detected, and represent a threat for local poultry and for the Houbara Bustard (Chlamydotis macqueenii) conservation breeding programs. To assess the presence, transmission, and maintenance of these viruses in such environments, 4,521 individuals from six resident and peridomestic wild bird species were sampled in the vicinity of two Houbara Bustard conservation breeding centers, and tested for AIV and NDV using real-time reverse transcriptase PCR and an enzyme-linked immunosorbent assay. A limited exposure to AIV was reflected in a virus prevalence below 0.4% and a serologic prevalence of 0.6%, and a moderate circulation of NDV was indicated by a virus prevalence of 0.9% and a serologic prevalence of 18.9% in the targeted peridomestic wild birds, suggesting different epidemiological roles for each taxa. Thus, some peridomestic species could actively participate in the epidemiological cycle of NDV in arid environments such as the UAE, challenging the conceptual epidemiologic framework centered on the involvement of waterfowl and shorebirds.

Evolution of overactive bladder symptoms in genital prolapse surgery / Evolución de los síntomas de vejiga hiperactiva tras la cirugía de prolapso genital

Objective: In the present study, we evaluated the development of symptoms of overactive bladder in women undergoing surgery for pelvic organ prolapse. Material and methods: We performed an epidemiological, longitudinal, prospective study involving 39 gynecological centers in Spain. The study population included women with symptomatic pelvic organ prolapse who were candidates for vaginal surgery. Patients attended a baseline visit (before surgery) and another visit 1 year after surgery. Pelvic organ prolapse was graded according to the Baden-Walker classification and the Pelvic Organ Prolapse Questionnaire. Urgency, urge incontinence, frequency, and nocturia were evaluated separately. Patients filled out the Epidemiology of Prolapse and Incontinence Questionnaire and the International Consultation on Incontinence Questionnaire. Results: The final sample included 360 women (319 were evaluable at the postsurgery visit). At baseline 58.3%, 38.3%, and 47.2% of women reported urgency, urge urinary incontinence, and frequency, respectively, whereas after surgery these symptoms were reported by 34.5%, 17.2%, and 19.7% (p < 0.001). The percentage of patients reporting nocturia also decreased after surgery from 59.4% to 56.4%, although the difference was not statistically significant. Furthermore, 19.8%, 9.8%, and 7.6% of patients, respectively, reported de novo urgency, urge urinary incontinence, and frequency after surgery. Patients with persistence of postsurgical urge urinary incontinence had significantly higher scores in the International Consultation on Incontinence Questionnaire at baseline. Conclusions: In general, a reduction in overactive bladder symptoms was observed in women undergoing surgical treatment for pelvic organ prolapse

Objetivo: el presente estudio pretende evaluar la evolución de síntomas de vejiga hiperactiva en mujeres intervenidas de prolapso de órganos pélvicos. Material y métodos: estudio epidemiológico, longitudinal, prospectivo, en 39 servicios de ginecología de España, que incluyó mujeres con prolapso de órganos pélvicos sintomático candidatas a cirugía de prolapso por vía vaginal. Se realizó una visita previa y una de seguimiento al año de la cirugía. Se evaluó el prolapso de órganos pélvicos mediante la clasificación de Baden y el cuestionario de prolapso de órganos pélvicos. Las pacientes cumplimentaron el cuestionario de epidemiología de prolapso e incontinencia y el de incontinencia urinaria. Se evaluaron los síntomas de urgencia, incontinencia urinaria de urgencia, frecuencia y nocturia. Resultados: se incluyeron 360 mujeres (319 evaluables en la revisión poscirugía). En la evaluación basal el 58,3%, 38,3% y 47,2% de las mujeres refirieron urgencia, incontinencia urinaria de urgencia y frecuencia, mientras que en la visita poscirugía únicamente el 34,5%, 17,2% y 19,7% las presentaban (p < 0,001). El porcentaje de pacientes que presentaban nocturia también se redujo, del 59,4% al 56,4%, aunque no de forma estadísticamente significativa. Un 19,8%, 9,8% y 7,6% de las pacientes presentaron urgencia, incontinencia urinaria de urgencia y frecuencia de novo tras la cirugía, respectivamente. Las pacientes con persistencia de incontinencia urinaria de urgencia poscirugía presentaron puntuaciones significativamente más elevadas en el cuestionario de incontinencia urinaria en la visita basal. Conclusiones: en general, se observa una reducción de los síntomas de vejiga hiperactiva en mujeres sometidas a cirugía de prolapso de órganos pélvicos

Muscle cell identity requires Pax7-mediated lineage-specific DNA demethylation.

BACKGROUND: Skeletal muscle stem cells enable the formation, growth, maintenance, and regeneration of skeletal muscle throughout life. The regeneration process is compromised in several pathological conditions, and muscle progenitors derived from pluripotent stem cells have been suggested as a potential therapeutic source for tissue replacement. DNA methylation is an important epigenetic mechanism in the setting and maintenance of cellular identity, but its role in stem cell determination towards the myogenic lineage is unknown. Here we addressed the DNA methylation dynamics of the major genes orchestrating the myogenic determination and differentiation programs in embryonic stem (ES) cells, their Pax7-induced myogenic derivatives, and muscle stem cells in proliferating and differentiating conditions. RESULTS: Our data showed a common muscle-specific DNA demethylation signature required to acquire and maintain the muscle-cell identity. This specific-DNA demethylation is Pax7-mediated, and it is a prime event in muscle stem cells gene activation. Notably, downregulation of the demethylation-related enzyme Apobec2 in ES-derived myogenic precursors reduced myogenin-associated DNA demethylation and dramatically impaired the expression of differentiation markers and, ultimately, muscle differentiation. CONCLUSIONS: Our results underscore DNA demethylation as a key mechanism driving myogenesis and identify specific Pax7-mediated DNA demethylation signatures to acquire and maintain the muscle-cell identity. Additionally, we provide a panel of epigenetic markers for the efficient and safe generation of ES- and induced pluripotent stem cell (iPS)-derived myogenic progenitors for therapeutic applications.

Epigenetics override pro-inflammatory PTGS transcriptomic signature towards selective hyperactivation of PGE2 in colorectal cancer.

BACKGROUND: Misregulation of the PTGS (prostaglandin endoperoxide synthase, also known as cyclooxygenase or COX) pathway may lead to the accumulation of pro-inflammatory signals, which constitutes a hallmark of cancer. To get insight into the role of this signaling pathway in colorectal cancer (CRC), we have characterized the transcriptional and epigenetic landscapes of the PTGS pathway genes in normal and cancer cells. RESULTS: Data from four independent series of CRC patients (502 tumors including adenomas and carcinomas and 222 adjacent normal tissues) and two series of colon mucosae from 69 healthy donors have been included in the study. Gene expression was analyzed by real-time PCR and Affymetrix U219 arrays. DNA methylation was analyzed by bisulfite sequencing, dissociation curves, and HumanMethylation450K arrays. Most CRC patients show selective transcriptional deregulation of the enzymes involved in the synthesis of prostanoids and their receptors in both tumor and its adjacent mucosa. DNA methylation alterations exclusively affect the tumor tissue (both adenomas and carcinomas), redirecting the transcriptional deregulation to activation of prostaglandin E2 (PGE2) function and blockade of other biologically active prostaglandins. In particular, PTGIS, PTGER3, PTGFR, and AKR1B1 were hypermethylated in more than 40 % of all analyzed tumors. CONCLUSIONS: The transcriptional and epigenetic profiling of the PTGS pathway provides important clues on the biology of the tumor and its microenvironment. This analysis renders candidate markers with potential clinical applicability in risk assessment and early diagnosis and for the design of new therapeutic strategies.

Overlapping DNA methylation dynamics in mouse intestinal cell differentiation and early stages of malignant progression.

Mouse models of intestinal crypt cell differentiation and tumorigenesis have been used to characterize the molecular mechanisms underlying both processes. DNA methylation is a key epigenetic mark and plays an important role in cell identity and differentiation programs and cancer. To get insights into the dynamics of cell differentiation and malignant transformation we have compared the DNA methylation profiles along the mouse small intestine crypt and early stages of tumorigenesis. Genome-scale analysis of DNA methylation together with microarray gene expression have been applied to compare intestinal crypt stem cells (EphB2high), differentiated cells (EphB2negative), ApcMin/+ adenomas and the corresponding non-tumor adjacent tissue, together with small and large intestine samples and the colon cancer cell line CT26. Compared with late stages, small intestine crypt differentiation and early stages of tumorigenesis display few and relatively small changes in DNA methylation. Hypermethylated loci are largely shared by the two processes and affect the proximities of promoter and enhancer regions, with enrichment in genes associated with the intestinal stem cell signature and the PRC2 complex. The hypermethylation is progressive, with minute levels in differentiated cells, as compared with intestinal stem cells, and reaching full methylation in advanced stages. Hypomethylation shows different signatures in differentiation and cancer and is already present in the non-tumor tissue adjacent to the adenomas in ApcMin/+ mice, but at lower levels than advanced cancers. This study provides a reference framework to decipher the mechanisms driving mouse intestinal tumorigenesis and also the human counterpart.

Long range epigenetic silencing is a trans-species mechanism that results in cancer specific deregulation by overriding the chromatin domains of normal cells.

DNA methylation and chromatin remodeling are frequently implicated in the silencing of genes involved in carcinogenesis. Long Range Epigenetic Silencing (LRES) is a mechanism of gene inactivation that affects multiple contiguous CpG islands and has been described in different human cancer types. However, it is unknown whether there is a coordinated regulation of the genes embedded in these regions in normal cells and in early stages of tumor progression. To better characterize the molecular events associated with the regulation and remodeling of these regions we analyzed two regions undergoing LRES in human colon cancer in the mouse model. We demonstrate that LRES also occurs in murine cancer in vivo and mimics the molecular features of the human phenomenon, namely, downregulation of gene expression, acquisition of inactive histone marks, and DNA hypermethylation of specific CpG islands. The genes embedded in these regions showed a dynamic and autonomous regulation during mouse intestinal cell differentiation, indicating that, in the framework considered here, the coordinated regulation in LRES is restricted to cancer. Unexpectedly, benign adenomas in Apc(Min/+) mice showed overexpression of most of the genes affected by LRES in cancer, which suggests that the repressive remodeling of the region is a late event. Chromatin immunoprecipitation analysis of the transcriptional insulator CTCF in mouse colon cancer cells revealed disrupted chromatin domain boundaries as compared with normal cells. Malignant regression of cancer cells by in vitro differentiation resulted in partial reversion of LRES and gain of CTCF binding. We conclude that genes in LRES regions are plastically regulated in cell differentiation and hyperproliferation, but are constrained to a coordinated repression by abolishing boundaries and the autonomous regulation of chromatin domains in cancer cells.

Using high-sensitivity troponin T: the importance of the proper gold standard.

OBJECTIVE: The study objective was to determine how best to use high-sensitivity cardiac troponin T (hscTnT) to diagnose myocardial infarction. METHODS: A total of 358 patients presenting with acute coronary syndromes sampled at admission and 2, 4, and 6 to 8 hours. Both contemporary cardiac troponin T (cTnT) and hscTnT were measured. Patients were classified with conventional cTnT values by independent investigators. Myocardial infarction required a cTnT value ≥99th reference percentile and a ≥20% change. RESULTS: Seventy-nine patients had non-ST-segment elevation myocardial infarction, 105 patients had unstable angina, and 174 patients had nonacute coronary syndromes. A cTnT cutoff at the 10% coefficient of variation value missed 14.5% of infarctions. hscTnT had a sensitivity at admission of 89.9%, but specificity was only 75.1% because of elevations in 45.3% and 25.3% of those with unstable angina and nonacute coronary syndromes, respectively. The optimal value for myocardial infarction diagnosis with hscTnT was 25 ng/L at admission and 30 ng/L during serial sampling. All infarctions were diagnosed within 4 hours, with a time saving of 11 and 68 minutes compared with a cTnT value at the 99th reference percentile value and a cTnT value at a coefficient of variation of 10%. By using the 99th percentile of hsTnT plus a ≥20% change, 25 additional infarctions were identified. With these included, the optimal cutoff decreased to 12 ng/L at admission and 13 ng/L over time, but time to diagnosis increased. CONCLUSIONS: The gold standard used to diagnose myocardial infarction makes a major difference in the results. When myocardial infarction is diagnosed using hscTnT 99th percentile values with a 20% change, more are identified, diagnosis is delayed, and the optimal value for use is reduced.

Dynamics of bivalent chromatin domains upon drug induced reactivation and resilencing in cancer cells.

Epigenetic deregulation revealed by altered profiles of DNA methylation and histone modifications is a frequent event in cancer cells and results in abnormal patterns of gene expression. Cancer silenced genes constitute prime therapeutic targets and considerable progress has been made in the epigenetic characterization of the chromatin scenarios associated with their inactivation and drug induced reactivation. Despite these advances, the mechanisms involved in the maintenance or resetting of epigenetic states in both physiological and pharmacological situations are poorly known. To get insights into the dynamics of chromatin regulation upon drug-induced reactivation, we have investigated the epigenetic profiles of two chromosomal regions undergoing long range epigenetic silencing in colon cancer cells in time-course settings after exposure of cells to chromatin reactivating agents. The DNA methylation states and the balance between histone H3K4 methylation and H3K27 methylation marks clearly define groups of genes with alternative responses to therapy. We show that the expected epigenetic remodeling induced by the reactivating drugs, just achieves a transient disruption of the bivalent states, which overcome the treatment and restore the transcriptional silencing approximately four weeks after drug exposure. The interplay between DNA methylation and bivalent histone marks appears to configure a plastic but stable chromatin scenario that is fully restored in silenced genes after drug withdrawal. These data suggest that improvement of epigenetic therapies may be achieved by designing strategies with long lasting effects.

Dihydrofolate reductase amplification and sensitization to methotrexate of methotrexate-resistant colon cancer cells.

Gene amplification is one of the most frequent manifestations of genomic instability in human tumors and plays an important role in tumor progression and acquisition of drug resistance. To better understand the factors involved in acquired resistance to cytotoxic drugs via gene amplification, we have analyzed the structure and dynamics of dihydrofolate reductase (DHFR) gene amplification in HT29 cells treated with methotrexate (MTX). Analysis of the DHFR gene amplification process shows that the amplicon exhibits a complex structure that is consistently reproduced in independent treatments. The cytogenetic manifestation of the amplification in advanced stages of the treatment may be in the form of double minutes or as a homogeneously stained region. To get insights into the mechanisms of resistance, we have also investigated the sensitization to MTX of MTX-resistant cells after drug withdrawal and reexposure to MTX. Passive loss of the DHFR amplicon by withdrawal of the drug results in MTX-sensitive cells exhibiting a substantial reduction of their capacity or even an incapacity to generate resistance when submitted to a second cycle of MTX treatment. On a second round of drug administration, the resistant cells generate a different amplicon structure, suggesting that the formation of the amplicon as in the first cycle of treatment is not feasible. These results indicate that DHFR gene amplification is a "wear and tear" process in HT29 cells and that MTX-resistant cells may become responsive to a second round of treatment if left untreated during a sufficient period of time.

Prognostic significance of molecular classification of breast invasive ductal carcinoma.

OBJECTIVE: Breast carcinoma classification has dramatically changed in recent years following application of molecular techniques. Immunohistochemistry can help select patients for different therapies. The objective of the present report is to determine the prognostic influence of the molecular classification of breast carcinoma with immunohistochemistry. MATERIALS AND METHODS: We have retrospectively selected a cohort of 257 patients with invasive ductal carcinoma NOS diagnosed and treated in the same hospital between 1997 and 2000. We have classified the cases in four tumor types according to the immunohistochemical expression of several markers, as luminal A tumors [estrogen and/or progesterone receptors (ER/PE) positive and Her-2 negative]; luminal B tumors (ER/PR positive and Her-2 positive); Her-2 positive tumors (ER/PR negative with Her-2 positive); and triple negative phenotype (all markers negative). RESULTS: In our series, 116 patients had tumors of luminal A type (47.93%); 67 (27.68%) were luminal type B; 33 (13.63%) were Her2 positive; and 26 (10.74%) were triple negative. The recurrence rate was 19% for luminal type A tumors, 25.4% for luminal type B, 39.4% for Her2 positive and 30.8% for triple negative lesions. The mean relapse free survival was 79.07, 73.07, 64.3 and 83,5 months for luminal A and B, Her-2 and triple negative lesions, respectively. Mortality rate reached 11.2% for luminal A tumors compared with 19.4, 33.3 and 26.9% for luminal B, Her2 and triple negative tumors, respectively. The mean overall survival for these groups was 88.42, 81.41, 77.62 and 93.6 months. CONCLUSION: Molecular classification with immunohistochemistry behaves as a significant prognosticator for breast invasive ductal carcinoma in our series of patients. The worse prognosis observed for Her2 expressing lesions may have changed after trastuzumab use.

Bivalent domains enforce transcriptional memory of DNA methylated genes in cancer cells.

Silencing of multiple cancer-related genes is associated with de novo methylation of linked CpG islands. Additionally, bivalent histone modification profiles characterized by the juxtaposition of active and inactive histone marks have been observed in genes that become hypermethylated in cancer. It is unknown how these ambiguous epigenetic states are maintained and how they interrelate with adjacent genomic regions with different epigenetic landscapes. Here, we present the analysis of a set of neighboring genes, including many frequently silenced in colon cancer cells, in a chromosomal region at 5q35.2 spanning 1.25 Mb. Promoter DNA methylation occurs only at genes maintained at a low transcriptional state and is characterized by the presence of bivalent histone marks, namely trimethylation of lysines 4 and 27 in histone 3. Chemically induced hyperacetylation and DNA demethylation lead to up-regulation of silenced genes in this locus yet do not resolve bivalent domains into a domain-wide active chromatin conformation. In contrast, active genes in the region become down-regulated after drug treatment, accompanied by a partial loss of chromatin domain boundaries and spreading of the inactive histone mark trimethylated lysine 27 in histone 3. Our results demonstrate that bivalent domains mark the promoters of genes that will become DNA methylated in adult tumor cells to enforce transcriptional silence. These bivalent domains not only remain upon drug induced gene reactivation, but also spread over adjacent CpG islands. These results may have important implications in understanding and managing epigenetic therapies of cancer.

Genome-wide tracking of unmethylated DNA Alu repeats in normal and cancer cells.

Methylation of the cytosine is the most frequent epigenetic modification of DNA in mammalian cells. In humans, most of the methylated cytosines are found in CpG-rich sequences within tandem and interspersed repeats that make up to 45% of the human genome, being Alu repeats the most common family. Demethylation of Alu elements occurs in aging and cancer processes and has been associated with gene reactivation and genomic instability. By targeting the unmethylated SmaI site within the Alu sequence as a surrogate marker, we have quantified and identified unmethylated Alu elements on the genomic scale. Normal colon epithelial cells contain in average 25 486 +/- 10 157 unmethylated Alu's per haploid genome, while in tumor cells this figure is 41 995 +/- 17 187 (P = 0.004). There is an inverse relationship in Alu families with respect to their age and methylation status: the youngest elements exhibit the highest prevalence of the SmaI site (AluY: 42%; AluS: 18%, AluJ: 5%) but the lower rates of unmethylation (AluY: 1.65%; AluS: 3.1%, AluJ: 12%). Data are consistent with a stronger silencing pressure on the youngest repetitive elements, which are closer to genes. Further insights into the functional implications of atypical unmethylation states in Alu elements will surely contribute to decipher genomic organization and gene regulation in complex organisms.

Endometriosis infiltrativa profunda / Deeply infiltrating endometriosis

Presentamos el caso de una paciente con endometriosis infiltrativa profunda, previamente intervenida de endometriosis ovárica, y su manejo posterior. La clínica de la paciente fue el indicador principal de la progresión de la enfermedad y las pruebas de imagen complementarias fueron imprescindibles para el diagnóstico de certeza y la orientación del correcto abordaje quirúrgico

We present a case of deeply infiltrating endometriosis and describe its management. The patient had previously undergone surgery for ovarian endometriosis. The patient's symptoms were the main indicator of disease progression and imaging techniques were essential for accurate diagnosis and surgical management

Selective pelvic arterial embolization in the management of obstetric hemorrhage.

OBJECTIVE: Retrospective evaluation of pelvic arterial embolization for the treatment of severe post-partum hemorrhage. METHODS: Data were collected, from our departmental clinical records, on all patients with life-threatening post-partum hemorrhage managed with arterial embolization between January 2001 and December 2003. RESULTS: During the period analyzed, there were 29,119 deliveries in our institution. Of these, 27 patients underwent pelvic arterial embolization to control severe hemorrhaging despite conservative management. Of the 27 patients, 22 (81.5%) had a vaginal delivery and 5 had a caesarean section. The major indication for embolization was uterine atony (15 women). Disseminated intravascular coagulation developed in 20 cases (74.1%). There were eight cases (29.6%) who underwent hysterectomy, seven of them pre-embolization. The most frequent vessel embolized was the uterine artery (13 cases; 38.3%). One patient (3.7%) presented complications related to the procedure. The success rate was 96.3%. CONCLUSION: Pelvic arterial embolization is a good therapeutic choice for severe post-partum hemorrhage refractory to conservative treatment measures.

Hypermethylation of the prostacyclin synthase (PTGIS) promoter is a frequent event in colorectal cancer and associated with aneuploidy.

Inactivation of specific tumor suppressor genes by transcriptional silencing associated with hypermethylation of the promoter is a common event in cancer. We have applied the amplification of intermethylated sites (AIMS) technique to a 100 human colorectal cancers and seven cell lines to identify recurrent alterations that may unveil silenced tumor suppressor genes. Bisulfite sequencing was used to confirm differential DNA methylation results. Gene expression analysis was performed by real-time RT-PCR. An AIMS band recurrently displayed in tumors but not in normal tissues was isolated and identified as part of the CpG island of the prostacyclin synthase (PTGIS) gene promoter. PTGIS promoter was hypermethylated in 43 out of 100 colorectal cancers and in all cell lines. Bisulfite sequencing and clonal analysis confirmed the results obtained by AIMS and demonstrated biallelic hypermethylation of PTGIS promoter. Hypermethylation of the PTGIS promoter was associated with diminished gene expression, that was restored after treatment with demethylating and histone deacetylases inhibitor agents. PTGIS hypermethylation was associated with aneuploidy and p53 mutations. In the adjusted model, PTGIS methylation, but not p53 mutation, maintained the association with aneuploidy. We conclude that epigenetic inactivation of the PTGIS gene is a recurrent alteration in colorectal carcinogenesis.

Association between C-reactive protein levels and N-terminal pro-B-type natriuretic peptide in pre-dialysis patients.

BACKGROUND: The prevalence of inflammation is high among patients with chronic renal failure but the reason of inflammation is unclear. We test the hypothesis that inflammation in chronic renal failure could be the consequence of an increased left-ventricular wall tension related to ventricular dysfunction, hypervolemia or both. METHODS: For assessing left-ventricular filling pressure, plasma level of N-terminal pro-B-type natriuretic peptide (N-BNP) was used, as B-type natriuretic peptide is secreted from the cardiac ventricles in response to increased wall tension. N-BNP levels and C-reactive protein (CRP) were measured on the same day in 75 pre-dialysis patients. A previous history of cardiomiopathy with systolic dysfunction was present in 27 (36%) of them. RESULTS: The levels of N-BNP were not normally distributed (mean: 2,589 +/- 4,514 pg/ml; median: 789 pg/ml). The distribution of CRP levels was also not normal (mean: 15 +/- 27 mg/l; median: 5 mg/l). Both parameters correlated significantly (r: 0.41; p < 0.005). N-BNP was higher (p < 0.001) in patients with known ventricular dysfunction. Excluding these patients, the correlation between N-BNP and CRP was stronger (r: 0.88; p < 0.001). Univariate analysis in these patients without known cardiomyopathy showed that N-BNP levels also correlated with systolic and diastolic blood pressure (r: 0.54; p < 0.005) and inversely with creatinine clearance (r: -0.43; p < 0.01), serum albumin (r: 0.6; p < 0.001) and hemoglobin levels (r: 0.37; p < 0.05). CRP levels correlated significantly (p < 0.01) with the same parameters as N-BNP in univariate analysis. However, in multiple stepwise regression analysis in which CRP was the dependent variable, only the association with N-BNP remained significant (r: 0.87; p < 0.001). CONCLUSIONS: Our results suggest a link between left-ventricular filling pressure and inflammation in patients with advanced renal insufficiency. The importance of strict volume control in these patients, in order to reduce left-ventricular pressure and therefore inflammation, should be considered.