Epigenetic Evaluation of N-(2-hydroxyphenyl)-2-Propylpentanamide, a Valproic Acid Aryl Derivative with Activity Against HeLa Cells.

BACKGROUND: Valproic acid (VPA) is an HDAC inhibitor (HDACI) with an anticancer activity, but is hepatotoxic. N-(2-hydroxyphenyl)-2-propylpentanamide (o-OH-VPA) is a VPA aryl derivative designed in silico as a selective inhibitor of HDAC8 with biological properties against HeLa, rhabdomyosarcoma and breast cancer cell cultures. OBJECTIVE: We studied the epigenetic mechanism of o-OH-VPA as an HDACI and evaluated whether it was toxic to normal cells. METHODS: HeLa cells and primary human fibroblasts were used for this study as carcinogenic and normal cells, respectively. Cell survival was evaluated by MTT assay, whereas viability and doubling time were determined by the Trypan-blue method. HDAC activity was tested using the colorimetric HDAC activity assay. The expression of p21 was analyzed by PCR and HDAC8 expression was also evaluated by real-time PCR. Cell cycle and caspase-3 activity were analyzed by flow cytometry and caspase-3 colorimetric assay, respectively. RESULTS: o-OH-VPA (IC50 = 0.1 mM) was fifty-eight times more effective than VPA (IC50 = 5.8 mM) to reduce HeLa cell survival. Furthermore, o-OH-VPA increased the doubling time of HeLa cells by 33% with respect to the control. o-OH-VPA acted as HDACI in HeLa cells without affecting the HDAC8 expression, arresting the cell cycle of HeLa cells in the G0/G1 phase due to the increase in p21 expression with the inhibition of caspase-3 activity without exhibiting toxicity toward normal cells. CONCLUSION: Our results revealed that o-OH-VPA is an HDACI with a selective effect against HeLa cells but without the known toxicity exerted by most pan-HDACIs on normal cells.

Public insurance program impact on catastrophic health expenditure on acute myocardial infarction.

OBJECTIVE: ST-segment elevation myocardial infarction (STEMI) has an important economic burden that poised the urgent need to evaluate its catastrophic medical expense. This study evaluates the first 5 years of the national health initiative called Popular Insurance (PI) at the National Institute of Cardiology in Mexico. STUDY DESIGN: Retrospective data analysis. METHODS: STEMI patients with (n=317) and without (n=260) PI were selected. Analysed variables included socio-economical context, management care, cost evaluation and three outcomes (mortality, hospital readmission and therapeutic adherence). Descriptive statistical analyses, Kaplan-Meier survival and Support Vector Machine models were used accordingly. RESULTS: Treatment costs were higher for PI-covered individuals (P=0.022) and only 1.89% of them remained in debt, in contrast to 16.15% of those without PI. Statistically significant differences were found in relation to days in hospital wards (P<0.001), imaging studies (P<0.001) and surgical materials (P=0.04). Survival analysis (P=0.44) and therapeutic adherence (P=0.38) showed no differences. Hospital readmission was predicted with an 81.97% accuracy. The most important predictive variables included were stent type, number of days at the coronary care unit and hospital wards. CONCLUSIONS: The PI has proven to be a successful program where no differences were found in terms of health care and survival, whereas it provides timely financial support for families facing catastrophic health challenging events.

Improving risk assessment for post-surgical low cardiac output syndrome in patients without severely reduced ejection fraction undergoing open aortic valve replacement. The role of global longitudinal strain and right ventricular free wall strain.

Low cardiac output syndrome (LCOS) after surgical aortic valve replacement (SAVR) is related to increased mortality and treatment related costs. We aimed to evaluate whether echocardiography-derived left ventricular global longitudinal strain (LV-GLS) relates to the occurrence of postoperative LCOS in patients undergoing SAVR. We prospectively enrolled 75 patients with symptomatic severe aortic stenosis, left ventricular ejection fraction (LVEF) >40%, NYHA Class

Phylogenetic Relationships within Phyllophaga Harris (sensu lato) (Coleoptera: Melolonthidae, Melolonthinae) with Emphasis on Listrochelus Blanchard.

As partial results of a long-term project for the revision of the supraspecific classification of the American Melolonthini, using phylogenetic methods, interesting information on the relationships between the subgeneric groups of Phyllophaga Harris proposed by Saylor were obtained. The genus Listrochelus was described by Blanchard in 1851. However, in 1940, Saylor reduced it as a subgenus of Phyllophaga. The objective of the present study is to confirm the monophyly of Listrochelus by means of a phylogenetic analysis. A total of 132 species were analyzed; 31 species of them belong to Listrochelus, 76 are from other groups of Phyllophaga, and 25 species are from the outgroup. A morphological matrix with 281 characters was codified. A traditional search with 1000 iterations was performed in TNT. Branch support was investigated using the bootstrap method. Parsimony analysis resulted in ten equally parsimonious trees. The topology obtained in the strict consensus tree shows that the limits of Listrochelus are very clear (bootstrap 99%), supported by five synapomorphies and a combination of eight character states that are not exclusive to the group. Based on the hypothesis obtained, the restitution of the genus Listrochelus is proposed.

Green tea compound epigallo-catechin-3-gallate (EGCG) increases neuronal survival in adult hippocampal neurogenesis in vivo and in vitro.

Epigallo-catechin-3-gallate (EGCG), found in the leaves of Camellia sinensis (green tea), has antioxidant- and scavenger-functions and acts neuroprotectively. It has been publicized as anti-aging remedy but data on potential cellular mechanisms are scarce. Recent studies claimed that EGCG specifically promotes neural precursor cell proliferation in the dentate gyrus of C57Bl/6 mice, without changes at the level of immature and mature new neurons. We here analyzed the effects of EGCG on adult hippocampal neurogenesis in male Balb/C mice and saw a different pattern. Two weeks of treatment with EGCG (0, 0.625, 1.25, 2.5, 5 and 10mg/kg) showed a dose-response curve that peaked at 2.5mg/kg of EGCG with significantly increased cell survival without affecting cell proliferation but decreasing apoptotic cells. Also, EGCG increased the population of doublecortin-(DCX)-expressing cells that comprises the late intermediate progenitor cells (type-2b and -3) as well as immature neurons. After EGCG treatment, the young DCX-positive neurons showed more elaborated dendritic trees. EGCG also significantly increased net neurogenesis in the adult hippocampus and increased the hippocampal levels of phospho-Akt. Ex vivo, EGCG exerted a direct effect on survival and neuronal differentiation of adult hippocampal precursor cells, which was absent, when PI3K, a protein upstream of Akt, was blocked. Our results thus support a pro-survival and a pro-neurogenic role of EGCG. In the context of the conflicting published results, however, potential genetic modifiers must be assumed. These might help to explain the overall variability of study results with EGCG. Our data do indicate, however, that natural compounds such as EGCG can in principle modulate brain plasticity.

Interethnic variation of the MMP-9 microsatellite in Amerindian and Mexican Mestizo populations: considerations for genetic association studies.

We studied the interethnic variation of the MMP-9 microsatellite in the Mestizo and Amerindian populations using blood samples collected from 435 healthy unrelated individuals from the Central Valley of Mexico. DNA samples were genotyped using the -90 (CA)12-27 repeat near the MMP transcriptional start site using capillary electrophoresis. Our data were compared with those from African, Asian, and European populations (N = 729). Both Mestizo and Amerindian populations were in Hardy-Weinberg equilibrium (P ≥ 0.05). However, strong genetic heterogeneity was found within the Mestizo population (94%, P ≤ 0.0001), which exhibited the highest frequency of Amerindian, African, and European alleles. Likewise, Amerindians showed 6.7% variation among populations (P ≤ 0.0001), suggesting a genetic substructure potentially associated with linguistic affiliations. These findings were corroborated with principal component and population differentiation analyses, which showed relative proximity among the Mestizos and their historical parental populations: Asian (FST ≥ 0.05), European (FST ≥ 0.09), and African (FST ≥ 0.02). Nevertheless, important differences were found between Mestizo and Nahuas (P ≤ 0.0001), and between Mestizo and Me'Phaas (P ≤ 0.0001). These findings highlight the importance of determining local-specific patterns to establish the population variability of MMP-9 and other polymorphic markers. Validation of candidate markers is critical to identifying risk factors; however, this depends on knowledge of population genetic variation, which increases the possibility of finding true causative variants. We also show that dissimilar ethnic backgrounds might lead to spurious associations. Our study provides useful considerations for greater accuracy and robustness in future genetic association studies.

Rat embryo quality and production efficiency are dependent on gonadotrophin dose in superovulatory treatments.

The present study was performed to optimize a superovulation protocol in rats in order to produce a large number of good-quality embryos suitable to develop rat embryonic stem (rES) cells. We first evaluated the ovulation kinetics of three rat strains: Wistar, Fisher and ACI/N. Animals (n=30 per strain) were treated with 50 IU of pregnant mare serum gonadotrophin (PMSG), and ovulation was induced with 50 IU of human chorionic gonadotrophin (hCG) 50 h apart. Next, we evaluated the dose-response curves of PMSG and hCG in Wistar rats in order to obtain the highest number of embryos. The parameters evaluated for superovulation efficiency were: percentage of mated females, percentage of pregnant females and the average number of embryos collected per female. The results of these experiments suggested that the best dose combination was 50 IU for each hormone. Subsequent experiments, again with Wistar rats, were designed to test which of four hormonal combination treatments (30/30, 30/50, 50/30, and 50/50 IU of PMSG/hCG) will produce the largest numbers of good-quality embryos. Embryo quality was evaluated by embryo development uniformity, embryo morphology, embryo survival in an in vitro culture and embryo ability to generate rES-like cells. Results from these experiments showed that 30/50 IU of PMSG/hCG was the treatment that induced the best embryo quality. In conclusion, our results indicated that, in Wistar rats, the most appropriate hormonal combination dose for superovulation protocols with high number of good-quality embryos was 30 IU of PMSG and 50 IU of hCG given 50 h apart. We are performing further studies with rES-like cells produced with the present methodology to evaluate if they are able to participate in the production of germ-line chimeras.

[Fast pathway for heart surgery]. / Vía rápida para cirugía cardiaca.

The increasing demographic and life expectancy rates, together with the vertiginous technological development during the last two decades, have raised the number of cardiac patients requiring surgical treatment. Therefore, several institutions have been forced to give priority to advanced or more serious cases and to postpone those that do not demand an urgent surgery. This analysis was made from June 15, 1999 to June 15, 2000 and demonstrates the results obtained from maintaining a fast track at the National Institute of Cardiology "Ignacio Chávez". This has favored the practice of surgeries in cases with noncomplex pathologies, null mortality, and low morbidity rates. The analysis also compares results from patients who had been subjected to surgery for the same type of pathologies but following the usual course of admittance and surgery scheduling, proving to be cost-efficient. It also demonstrates the high incidence of inter-auricular communication and the Von Willebrand disease that exist in our environment. Based on the results, we propose to maintain and increase this type of surgeries, addressing their limitations.

[Mechanical reperfusion in acute myocardial infarction]. / Reperfusión mecánica en el infarto agudo del miocardio.

Nowadays, it has been demonstrated that reperfusion therapy in acute myocardial infarction diminishes mortality and improves ventricular function. Primary percutaneous transluminal coronary angioplasty (ACTPp) was described in 1983 by Hartzler as an alternative treatment in acute myocardial infarction. The results of this procedure were considered excellent. In 1996 the committee built up by the ACC/AHA published in the 1999 updated JACC's issue, a statement of the position that occupied at that moment primary angioplasty in the treatment of acute myocardial infarction. This committee pointed that this therapy showed to be an excellent alternative compared with the use of thrombolytic agents in acute myocardial infarction with ST elevation, patients who presented < 12 hs of the onset of symptoms according to the following conditions: 1. In less time, 2. Supported by trained staff, and 3. In an experience cath lab. This communication was introduced prior to the use of interventionist therapy with stents with or without the utilization of glycoproteins inhibitors. The analysis of results of different comparative studies of ACTPp with or without stents vs the use with or without II/IIIa glycoproteins inhibitors, was also included in this communication; among them it stands out the epistent which demonstrated less events as far as 6 months and was statistical significant in patients treated with Stent and abciximad. In conclusion we believe that ACTPp with Stent and glycoprotein inhibitors offers better results than those obtained from a combined therapy with thrombolytic agents and IIb/IIIa, however, we must recognise that there are limitations when performing angioplasty, as there is a lack of an efficient 24 hr. cath lab.

In vivo gene transfer to dopamine neurons of rat substantia nigra via the high-affinity neurotensin receptor.

BACKGROUND: Recently, we synthesized a nonviral gene vector capable of transfecting cell lines taking advantage of neurotensin (NT) internalization. The vector is NT cross-linked with poly-L-lysine, to which a plasmid DNA was bound to form a complex (NT-polyplex). Nigral dopamine neurons are able to internalize NT, thus representing a target for gene transfer via NT-polyplex. This hypothesis was tested here using reporter genes encoding green fluorescent protein or chloramphenicol acetyl transferase. MATERIALS AND METHODS: NT-polyplex was injected into the substantia nigra. Double immunofluorescence labeling was used to reveal the cell type involved in the propidium iodide-labeled polyplex internalization and reporter gene expression. RESULTS: Polyplex internalization was observed within dopamine neurons but not within glial cells, and was prevented by both hypertonic sucrose solution and SR-48692, a selective nonpeptide antagonist of NT receptors. Reporter gene expression was observed in dopamine neurons from 48 hr up to 15 days after NT-polyplex injection, and was prevented by SR-48692. However, no expression was seen when the NT-polyplex was injected into the ansiform lobule of the cerebellum, which contains low- but not high-affinity NT receptors. Neither internalization nor expression was observed in cultured glial cells, despite the NT-polyplex binding to those cells that was prevented by levocabastine, a low-affinity NT receptor antagonist. CONCLUSIONS: These results suggest that high-affinity NT receptors mediate the uptake of NT-polyplex with the subsequent reporter gene expression in vivo. NT polyfection may be used to transfer genes of physiologic interest to nigrostriatal dopamine neurons, and to produce transgenic animal models of dopamine-related diseases.

Prognostic implications of right atrial ischemic dysfunction in patients with biventricular inferior infarction: transesophageal echocardiographic analysis.

In order to determine the effect of right atrial dysfunction on clinical outcome, six patients with inferior myocardial infarction with extension to right ventricle and right atrium involving only obstructions of the right coronary artery were examined with transesophageal echocardiography (TEE) at the time of the event. Five of the patients were reexamined 15 to 55 months later. Two patients underwent thrombolysis and maintained ratios of right-to-left ventricular diameters of less than 1, as well as normal convexity of the interatrial septum. One patient had spontaneous reperfusion of the right coronary artery, reduction in right ventricular diameter, and normalization of interatrial septum. Another patient underwent delayed angioplasty and manifested a diminished wall movement score (WMS) in the follow-up echocardiogram. One patient died during his first hospitalization with significant right ventricular dilatation, inverted convexity of the interatrial septum, and right atrial thrombosis. The last patient died during follow-up with right ventricular dilatation, increased WMS, right atrial akinesis, and inverted interatrial convexity. Serial TEE examination of patients with infarction of the left ventricular inferior wall is a safe technique for determining the degree of the extension of the ischemic process to the right chambers.

Left atrial thrombosis after percutaneous mitral valvuloplasty: resolution with conservative management.

We report the detection of a thrombus 72 hours after mitral valvuloplasty through the use of the technique of Inoue. Images obtained by transesophageal echocardiography revealed its localization on the interatrial septum at the level of the transseptal puncture. Although the patient subsequently underwent surgery for the placement of a prosthetic valve in mitral position due to failure of the valvuloplasty, the thrombus resolved with conservative management.

[The benefit of a coronary stent in the reduction of major cardiac complications in acute coronary syndromes]. / Beneficio del stent coronario en la reducción de complicaciones cardiacas mayores en síndromes coronarios agudos.

To assess the incidence of in-hospital major adverse cardiac events (MACE), we analyzed 694 procedures in 613 consecutive patients during one year period. Patient population included 550 (79.2%) patients with unstable angina, 43 (6.2%) with stable angina and 101 (14.5%) with acute myocardial infarction. Elective percutaneous transluminal coronary angioplasty (PTCA) was performed in 593 (85.4%) patients, rescue PTCA in 7 (1%), and primary PTCA in 94 (13.5%). Angiographic lesion morphology was as follows: type A 30%; type B 58%; type C 12%. We compared patient population who received stent with PTCA-balloon only. Technical success was 95% and clinical success was achieved in 80% of the cases. Overall mortality was 1% in the stent group and 3% in the conventional PTCA. The incidence of MACE was 4% and 15.1% in the stent and angioplasty balloon groups respectively. We found a dramatic impact on reduction of the incidence of acute complications in the groups with stenting for unstable angina (p = 0.0001) and acute myocardial infarction (p = 0.0001). The major clinical advantage of stenting over balloon angioplasty was a lower need for repeated procedures.

[Translumbar coronary angiography and aortography in a patient with Takayasu's arteritis]. / Arteriografía coronaria y aortografía por vía translumbar en una paciente con arteritis de takayasu.

Since its introduction by Dos Santos in 1929, arterial angiography by translumbar percutaneous approach has suffered some transformations. Nowadays it has been replaced by other percutaneous approaches and it is indicated only when these routes of access have failed due to aortoiliac or subclavian arteries obstruction. This report presents a patient with Takayasu's Arteritis with severe peripheral arterial obstruction and unstable angina, who underwent coronary arteriography and aortography by translumbar approach. A review of this technique is made.

Neoadjuvant chemotherapy interferon for epidermoid penile cancer: a case report

Antecedentes. El carcinoma avanzado de pene es un tumor con un pronóstico malo con el tratamiento estándar por lo que la quimioterapia neoadyuvante se ha estado evaluando para mejorar la preservación del órgano y la supervivencia. Caso clínico. Paciente de 56 años visto en el Instituto Nacional de Cancerología en noviembre de 1993 siendo diagnosticado de cáncer de pene T3 N3 MO. El tratamiento consistió en cisplatino 100 mg/m2 cada 21 días por cinco ciclos alternados con metotrexato intravenoso a dosis de 250 mg/m2 más rescate con leucovorín oral a dosis de 10 mg/m2 cada ocho horas por seis dosis. El metotrexato y leucovorín se administraron cada 15 días en siete ocasiones. Además recibió interferón-alfa 4.5 x 10 6 U por vía subcutánea cada tercer día durante las 12 semanas del tratamiento. Resultados. El tratamiento fue bien tolerado alcanzando una respuesta clínica del 70 por ciento en el tumor primario y completa en los ganglios inguinales. Fue sometido a penectomía radical en abril de 1994 y actualmente está libre de enfermedad a los 61 meses de seguimiento. Conclusiones. Esta modalidad de quimioinmunoterapia parece ser muy activa en el cáncer de pene y debería evaluarse en un número mayor de pacientes con el fin de preservar el órgano e incrementar la supervivencia

Interventional cardiology in congenital heart disease.

Interventional procedures for congenital heart disease have evolved dramatically in the last decade. Beginning with pulmonary and aortic valvuloplasty, nowadays, stents and various devices are placed inside the heart and vessels to palliate or correct different congenital defects. The present communication summarizes the experience with interventional cardiology in congenital heart disease of the Instituto Nacional de Cardiología "Ignacio Chávez" during the last five years.

Percutaneous mitral commissurotomy.

Rheumatic fever is the most common cause of acquired heart disease in children and young adults worldwide, mainly in developing countries, and is the reason that a large number of patients with mitral stenosis will require some modality of treatment during their life span. The early therapeutic approach for this condition was solely surgical, but since the early 1960s, several balloon catheter interventional techniques have emerged as alternatives to surgical treatment. In 1976 Inoue described a novel single-balloon device designed specifically for percutaneous transvenous mitral commissurotomy (PMC), and in 1982 the first clinical application of Inoue's technique was successfully accomplished. Ever since several clinical trials established PMC as an effective and safe procedure for severe mitral stenosis in more than 30,000 patients worldwide, percutaneous balloon techniques have been considered the method of choice in selected patients (functional class >II, mitral valve area <1.5 cm2, and Wilkins' score <8) for several reasons. PMC is a nonsurgical method with results similar to those of surgical intervention, but without the unnecessary risks and complications of general anesthesia and extracorporeal circulation pump. Both PMC and surgical commissurotomy yield comparable acute and long-term results and similar restenosis rates. Moreover, PMC is the strategy of choice in symptomatic pregnant women with mitral stenosis, as well as in some cases with restenosis after a previous PMC. Patient selection and procedural technique are reviewed in detail.

Pericardium-covered stent for septal myocardial ablation in hypertrophic obstructive cardiomyopathy.

This report describes a patient with severe hypertrophic obstructive cardiomyopathy in New York Heart Association functional class III. Complete reduction of left ventricular outflow tract gradient was achieved by the selective occlusion of three target septal arteries with a pericardium-covered stent. The patient's in-hospital course was uneventful and has improved to functional class I.

Transjugular liver biopsy in 52 patients with an automated Trucut-type needle.

A three-year prospective study was conducted to assess the efficacy and safety of transjugular liver biopsy with a Trucut-type needle employing an automated device. Fifty-two consecutive patients (39 women and 13 men), ages 46.3+/-14.6 years, in whom percutaneous liver biopsy was contraindicated were included in the study. An 18-gauge transjugular hepatic needle with a 20-mm throw length, a cutting cannula at the distal end, and an automated trigger device on the proximal end was used. All procedures were performed under fluoroscopic control, and the following variables were assessed: (1) number of passes, (2) size and average number of the obtained fragments, (3) number of portal triads, and (4) adequacy of the specimen for histologic diagnosis. The procedure was successful in 49/52 patients, and all samples obtained were satisfactory for histologic analysis even when cirrhosis or bridging fibrosis were present. Mean biopsy specimen length was 1.7+/-0.88 cm; mean number of passes was 2.42+/-1.01, the mean number of biopsy fragments and portal triads per attempt were 2.5+/-1.01 and 6.24+/-3.18, respectively. No major complications were observed. Transjugular hepatic biopsy with this new cutting system is an effective and safe procedure in patients with contraindication for percutaneous liver biopsy.