Predictive and prognostic value of total tumor load in sentinel lymph nodes in breast cancer patients after neoadjuvant treatment using one-step nucleic acid amplification: the NEOVATTL study.

OBJECTIVE: To evaluate the predictive and prognostic value of total tumor load (TTL) in sentinel lymph nodes (SLNs) in patients with infiltrating breast cancer after neoadjuvant systemic therapy (NST). METHODS: This retrospective multicenter study used data from a Spanish Sentinel Lymph Node database. Patients underwent intraoperative SLN biopsy after NST. TTL was determined from whole nodes using a one-step nucleic acid amplification (OSNA) assay and defined as the total sum of CK19 mRNA copies in all positive SLNs. Cox-regression models identified independent predictive variables, which were incorporated into a nomogram to predict axillary non-SLN metastasis, and identified prognostic variables for incorporation into a disease-free survival (DFS) prognostic score. RESULTS: A total of 314 patients were included; most had no lymph node involvement prior to NST (cN0; 75.0% of patients). Most received chemotherapy with or without biologic therapy (91.7%), and 81 patients had a pathologic complete response. TTL was predictive of non-SLN involvement (area under the concentration curve = 0.87), and at a cut-off of 15,000 copies/µL had a negative predictive value of 90.5%. Nomogram parameters included log (TTL + 1), maximum tumor diameter and study-defined NST response. TTL was prognostic of disease recurrence and DFS at a cut-off of 25,000 copies/µL. After a 5-year follow-up, DFS was higher in patients with ≤ 25,000 copies/µL than those with > 25,000 (89.9% vs. 70.0%; p = 0.0017). CONCLUSIONS: TTL > 15,000 mRNA copies/µL was predictive of non-SLN involvement and TTL > 25,000 mRNA copies/µL was associated with a higher risk of disease recurrence in breast cancer patients who had received NST.

Obtención de plasma rico en plaquetas (PRP) en el Laboratorio de Terapia Celular para uso como herramienta terapéutica en medicina regenerativa / Obtaining platelet rich plasma (PRP) in the Cell Therapy Laboratory for use as a therapeutic tool in regenerative medicine

Platelet rich plasma (PRP) is used to speed up tissue repair. Despite its widespread use, the therapeutic application of PRP generates controversies in clinical results due to the variability in methods of obtaining the different preparations and differences between the components of different types of PRP, so it's recommended to mention the type of platelet preparation used. In this article, we describe technical and biologics characteristics of our platelet product, and we compare them to different commercial preparations described in order to validate their clinical use. Our results determine that the preparation can be considered a platelet rich plasma with biological activity in vivo and in vitro, which supports its use as a valid therapeutic tool, alternative to products currently available in Regenerative Medicine. (AU)

Post-exercise hypotension and skeletal muscle oxygenation is regulated by nitrate-reducing activity of oral bacteria.

Post-exercise hypotension (PEH) is a common physiological phenomenon leading to lower blood pressure after acute exercise, but it is not fully understood how this intriguing response occurs. This study investigated whether the nitrate-reducing activity of oral bacteria is a key mechanism to trigger PEH. Following a randomized, double blind and crossover design, twenty-three healthy individuals (15 males/8 females) completed two treadmill trials at moderate intensity. After exercise, participants rinsed their mouth with antibacterial mouthwash to inhibit the activity of oral bacteria or a placebo mouthwash. Blood pressure was measured before, 1h and 2 h after exercise. The microvascular response to a reactive hyperaemia test, as well as blood and salivary samples were taken before and 2 h after exercise to analyse nitrate and nitrite concentrations and the oral microbiome. As expected, systolic blood pressure (SBP) was lower (1 h: -5.2 ±â€¯1.0 mmHg; P < 0.001); 2 h: -3.8 ±â€¯1.1 mmHg, P = 0.005) after exercise compared to baseline in the placebo condition. This was accompanied by an increase of circulatory nitrite 2 h after exercise (2h: 100 ±â€¯13 nM) compared to baseline (59 ±â€¯9 nM; P = 0.013). Additionally, an increase in the peak of the tissue oxygenation index (TOI) during the reactive hyperaemia response was observed after exercise (86.1 ±â€¯0.6%) compared to baseline levels (84.8 ±â€¯0.5%; P = 0.010) in the placebo condition. On the other hand, the SBP-lowering effect of exercise was attenuated by 61% at 1 h in the recovery period, and it was fully attenuated 2 h after exercise with antibacterial mouthwash. This was associated with a lack of changes in circulatory nitrite (P > 0.05), and impaired microvascular response (peak TOI baseline: 85.1 ±â€¯3.1%; peak TOI post-exercise: 84.6 ±â€¯3.2%; P > 0.05). Diversity of oral bacteria did not change after exercise in any treatment. These findings show that nitrite synthesis by oral commensal bacteria is a key mechanism to induce the vascular response to exercise over the first period of recovery thereby promoting lower blood pressure and greater muscle oxygenation.

Estandarización de la técnica Real Time PCR para la detección molecular de la mutación V617F en el gen de JAK-2 en neoplasias mieloproliferativas crónicas / Standardization of the Real Time PCR technique for the molecular detection of the V617F mutation in the JAK-2 gene in chronic myeloproliferative neoplasms

V617F mutation in exon 14 of Janus Kinase 2 gene (jak-2) is used as a molecular marker for the diagnosis of Philadelphia negative myeloproliferative neoplasms (Phi-) such as Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Primary Myelofibrosis (MFP). To detect this mutation, we used conventional polymerase chain reaction technique (PCR), a simple and inexpensive technique, however, has some drawbacks that current technology allows to solve. During the last years, more sensitive molecular techniques have been incorporated in clinical practice to support the diagnosis, prognosis and follow-up of hematological patients. For its implementation in the clinical routine should be considered technical and economic aspects, so in this work, we evaluate the Real Time PCR technique as a diagnostic method for the detection of the Jak-2-V617F mutation, using in house primers design. Our result show that the technique implemented has a concordance index of 0.87 with the conventional PCR used in the molecular diagnosis of myeloproliferative neoplasms. In addition, it has the same specificity, greater sensitivity and, shorter execution time in relation to conventional PCR. The implementation of this diagnostic method in our Hospital is technically possible and commercially convenient. (AU)

Prevalence of bacterial contamination in platelet concentrates at the National Center of Blood Transfusion (Mexico).

OBJECTIVES: Most common bacterial sepsis associated with transfusion is caused by contaminated Platelet Concentrates (PC). The screening of PC to detect bacterial contamination is obligatory in Mexico, and it is carried out in quality control programs. In Mexico, the identification and molecular characterization of bacterial contaminants to detect contamination sources have not been implemented due to high costs; however, it is an actual current need. MATERIAL AND METHODS: One hundred PC were randomly selected and microbiologically analyzed. This sample size corresponds to 1% of the PC obtained by the National Center of Blood Transfusion (NCBT) in Mexico City according to the Official Mexican Standard NOM-253-SSA1-2012. Additionally, molecular biology tests were implemented in order to identify the possible contamination sources. RESULTS: Nine of the 100 PC analyzed (9%) showed bacterial contamination; analysis of the nucleotide sequences revealed the presence of characteristic microbiota from donor skin and soil. Diverse clonal relationship between the strains was identified in Staphylococcus epidermidis. CONCLUSION: Detection of contaminants associated with environmental and skin flora, shows the need to implement measures in the process of disinfecting skin at the site of phlebotomy and cleaning each of the areas involved in blood collection.

Razones por las que las personas donan o no donan sangre en Guatemala / Reasons why people donate or do not donate blood in Guatemala

Estudio descriptivo realizado para determinar la percepción que tienen los guatemaltecos y guatemaltecas sobre la donación voluntaria de sangre. Se utilizaron tres encuestas dirigidas a tres poblaciones diferentes: personas que si donan, personas que no donan (observadoras) y personas que fueron diferidas durante la donación . De las 518 personas que acudieron a las diferentes colectas de donación voluntaria, únicamente 378 (73%) participaron en las encuestas. Se logró establecer que las personas que donan se sienten motivadas a colaborar con los que necesitan los hemoderivados. Los que fueron diferidos se mostraron decepcionados por no haber logrado pasar el proceso previo a la donación. Los observadores sentían curiosidad por el proceso pero no mostraron disposición para donar, o para aceptar la responsabilidad de que su sangre fuera transfundida...(AU)

It is a descriptive study to determine the perception Guatemalans on voluntary blood donation. If people donate , people do not donate ( observer ) and people that were deferred during the donation , three surveys of three different populations were used. Of the 518 people who attended the various collections of voluntary donation, only 378 ( 73 %) participated in the polls. It was established that people who donate are motivated to collaborate with those who need blood products. Those who were deferred were disappointed not to have made it through the pre- donation process. The observers were curious about the process but did not show willingness to donate , or to accept responsibility for his blood was transfused...(AU)

[Frequency of minor histocompatibility antigens among Chilean blood donors]. / Frecuencia génica de antígenos menores de histocompatibilidad en la población chilena y estimación de sus efectos inmunológicos en el trasplante alogénico de progenitores hematopoyéticos.

BACKGROUND: Minor histocompatibility antigens (mHAgs) play a critical role in the immune responses associated with allogeneic stem cell transplantation, such as graft versus host disease (GVHD) and graft-versus-tumor (GVT). AIM: To determine the gene frequencies of the mHAgs HA-1, HA-2 and HA-8 in Chilean Blood Bank donors. MATERIAL AND METHODS: Blood from 192 blood donors was analyzed. The presence of haplotype HLA-A*02 was determined by flow cytometry. The frequency of mHAgs was determined by allele specific polymerase chain reaction in genomic DNA. RESULTS: Sixty one participants were carriers of the haplotype HLA-A*02. The relative allele frequency HA-1H was 45%, HA-Ir 55%, HA-2V 80.6%, HA-2M 19.4%, HA-8R 49.8% and HA-8P was 50.2%. Based on mHAgs disparity between HA-1, HA-2 or HA-8, the probability to generate a GVT response in HLA-A*02 individuals was 40%. CONCLUSIONS: The mHAgs frequency in Chilean population is under Hardy-Weinberg equilibrium and they are similar to those of other ethnic populations in the world.

Frecuencia génica de antígenos menores de histocompatibilidad en la población chilena y estimación de sus efectos inmunológicos en el trasplante alogénico de progenitores hematopoyéticos / Frequency of minor histocompatibility antigens among Chilean blood donors

Background: Minor histocompatibility antigens (mHAgs) play a critical role in the immune responses associated with allogeneic stem cell transplantation, such as graft versus host disease (GVHD) and graft-versus-tumor (GVT). Aim: To determine the gene frequencies of the mHAgs HA-1, HA-2 and HA-8 in Chilean Blood Bank donors. Material and Methods: Blood from 192 blood donors was analyzed. The presence of haplotype HLA-A*02 was determined by flow cytometry. The frequency of mHAgs was determined by allele specific polymerase chain reaction in genomic DNA. Results: Sixty one participants were carriers of the haplotype HLA-A*02. The relative allele frequency HA-1H was 45%, HA-Ir 55%, HA-2V 80.6%, HA-2M 19.4%, HA-8R 49.8% and HA-8P was 50.2%. Based on mHAgs disparity between HA-1, HA-2 or HA-8, the probability to generate a GVT response in HLA-A*02 individuals was 40%. Conclusions: The mHAgs frequency in Chilean population is under Hardy-Weinberg equilibrium and they are similar to those of other ethnic populations in the world.

Bilateral trans-humeral arm transplantation: result at 2 years.

Unilateral and bilateral hand transplantations have been performed worldwide with good mid-term functional results. An above-elbow bilateral transplantation was performed in a 29-year-old male patient from a fully HLA-mismatched donor. Alemtuzumab induction and steroid-free maintenance immunosuppression with tacrolimus and mycophenolate was used. Due to acute rejection, steroids were introduced at 6 months. Three acute rejection episodes occurred, one treated with alemtuzumab. New-onset diabetes after transplant, dyslipemia and worsening of previous high blood pressure required treatment. At 26 months post-transplantation, the patient has excellent elbow active movement, active flexion and extension of the thumb and fingers, useful sensation and a gainful job. Based on the functional results of the case reported, bilateral trans-humeral transplantation could be a viable treatment for selected bilateral above-elbow amputees.

Respuesta inmune contra antígenos menores de histocompatibilidad en el trasplante alogénico de progenitores hematopoyéticos / Immunologycal response under antigens hematopoietic stem cell transplantation

Patients who receive allo hematopoietic stem cell transplantation (SCT) could develop graft versus host disease and/or graft versus tumour effect. These immunological responses can happen even with perfect fully HLA matched haematopoietic stem cells. Moreover, the engraftment of the donor’s cells depends on the immunological conditions of both donor and recipient. The development of alloreactivity occurs in the context of the polymorphisms of the human genome, these genomic differences results in proteins with antigenic properties which trigger immune responses. Considering this, the SCT is a powerful tool to heal the patient disease, because all of them become chimeras. In other words, into individuals with two different genomic sets, which will develop a strong immunological response that cannot exist in natural conditions.

Programa de higiene oral para niños y jóvenes con discapacidad congnitiva, psicomotora y del lenguaje / Oral hygiene program for children and young persons with cognitive, psychomotor and language disabilities

Objetivo: diseñar, implementar y evaluar un programa de higiene oral para niños y jóvenes con discapacidad cognitiva, psicomotora y de lenguaje. Materiales y métodos: este estudio es cuasi-experimental y su objetivo es mejorar la higiene bucal mediante la implementación de un programa que utiiza las estrategias de musicoterapia (Minueto y Rondó del divertimento en Re efecto Mozart) y tableros de comunicación, en una población de 41 niños y jóvenes etre 3 y 28 años con discapacidad cognitiva, psicomotora y del lenguaje, de la Corporación Seres Integrales Serin. Se hizo una revisión de las deficiencias de cada uno de los niños, se diseñó el programa, capacitación a padres y terapeutas, acondicionamiento de la planta física y acercamiento a la comunidad. Se realizó toma del índice inicial, actividades alusivas a la higiene bucal, toma de índice intermedio, refuerzo de actividades y posteriormente toma de índice final. Resultados: después de la implementación del nivel 0 de estimulación con niños con discapacidad, disminuyó el 33 por ciento del índice de higiene bucal; el nivel 1 disminuyó el 49 por ciento del índice de higiene oral; el nivel 2 disminuyó en un 56 por ciento; el nivel 3 disminuyó el 59 por ciento del índice de higiene oral, el nivel 4 disminuyó el 68 por ciento del índice de higiene oral y el nivel 5 disminuyó en un 70 por ciento del índice de higiene oral. Conclusión: programa de higiene oral donde se utilizaron estrategias tales como la musicoterapia y tableros de comunicación. La capacitación a padres y tutores favoreció la disminución del índice de higiene oral en niños y jóvenes con discapacidad cognitiva psicomotora y del lenguaje.

[Interobserver correlation in the interpretation of 99mTc-sestamibi SPECT in reperfused acute myocardial infarction]. / Correlación interobservadores en interpretación de SPECT de perfusión con 99mTc-sestamibi en infarto agudo de miocardio reperfundido.

INTRODUCTION AND OBJECTIVES: The correct interpretation of myocardial perfusion single photon emission computed tomography (SPECT) requires knowledge of the technique reproducibility. The objective was analyze the interobserver correlation of different experience in the interpretation of myocardial perfusion SPECT in patients following acute myocardial infarction (AMI) in order to improve the quality of our site. METHODS: Sixty cases (56 +/- 11 years, 87 % men) with transmural AMI who had recently undergone successful thrombolysis were included. Resting perfusion with (99m)Tc-sestamibi was performed at one week post-AMI. ANALYSIS: Semiquantitative interpretation using 17 segment-model by 2 independent specialists and 5 observers, was performed blindly. Left ventricular ejection fraction (LVEF) was measured with isotopic ventriculography one month after AMI, with a mean of 38 %. RESULTS: Using independent and then agreed on perfusion analysis, average involved segments/patient was 9.3 +/- 4 and the sum of severity 25 +/- 13. Readings of other observers ranged from 7 +/- 3.7 to 9.4 +/- 3.9 and 16.7 +/- 9.7 to 24.6 +/- 13, respectively, consistent with the reading of the specialists of between 0.779-0.871 (kappa: 0.565-0.741). There was no significant difference when the number of segments were analyzed in 40 % of the cases and for intensity in 60 % of them in more experienced observers. Correlation with consensus reading for the number of segments ranged from 0.84 to 0.94 and for severity from 0.79 to 0.89. Identification of culprit arteries was acceptable, with r values between 0.612 and 0.683 and kappas between 0.629 and 0.656. Correlation of the number of involved segments and severity with LVEF performed one month after AMI was 0.73 and 0.74, respectively. CONCLUSIONS: There was good correlation in the interpretation of myocardial perfusion SPECT, with a significantly better fit in more experienced observers. This academic exercise was also helpful in improving our residents' skills in cardiology.

Trasplantes de progenitores hematopoyéticos / Hematopoietic stem cell transplant

The stem cell transplantation (SCT) has improved the disease free survival of a great number of diseases. It began as an experimental procedure, used as last resource in terminally ill patients. Nowadays it is a proved tool that allow the patient receive high dose of chemoradiotherapy, without the bone marrow toxicity being a conditioning step. The haematological recovery follows the same mechanisms that the organisms have designed for this purpose. First, cryopreservation of progenitors stem cells (PSCs) is required, then these cells are re-infused into the blood. The PSCs find their own homing in the bone marrow, proliferate, differentiate and re establish the haematopoietic balance. The present revision gives some information about the different phase or a SCT, physiology background, indications and principals adverse effects.

Estudio de quimerismo: aplicación en pacientes con trasplante de progenitores hematopoyéticos / Chimerism study: application in patients with allogeneic progenitor cell transplantation

El trasplante alogénico de progenitores hematopoyéticos (TAPH) es una técnica que ha cambiado el pronóstico de muchas enfermedades hematológicas malignas y no malignas. En determinados tiempos post trasplante coexisten células hematológicas de receptor y dador, por lo cual el individuo posee dos sistemas hematopoyéticos. El término Quimera se utiliza para indicar el origen dual de las células hematológicas. Este análisis es indispensable para saber si existe prendimiento o rechazo del trasplante. La determinación de quimerismo se realiza mediante la amplificación por PCR de secuencias cortas repetidas tandem (STRs del ingles short tandem repeat sequence). Este examen es fácil de realizar, reproducible, altamente sensible y específico. El resultado del quimerismo es de vital importancia a la hora de tomar decisiones clínicas, sobre todo si se utilizan técnicas no mieloablativas de acondicionamiento, infusión de linfocitos del donante, o modificaciones en los protocolos de inmunosupresión, donde existe una lata variabilidad en el prendimiento del injerto y el desarrollo de enfermedad de injerto contra huésped (EICH) e injerto contra tumor (ICT).

Allogeneic progenitor cell transplantation (ALoPCT) is a procedure that has changed the prognosis of many malignant and non-malignant hematologic diseases. For a period of time post-trasplant, cellular subtypes from the donor and the host coexist, giving the patient two hematopoietic systems. The term Chimera is used to indicate the dual origin of blood cells. This analysis is important to determine whether engrafment or rejection has occurred. The determination of chimerism is based on PCR pf Short Tandem Repeat sequences (STRs). The PCR technique is easy to perform, reproducible, highly sensitive and specific. Chimerism determination helps to improve the clinical approach to the patient, specifically when non-mieloablative conditioning, lymphocyte donor infusion or modification of the immunosuppressive protocols have employed. All of these manipulations produce a high variability in engraftment, development of graft versus host disease (GVHD) and the likelihood to eliminate tumor cells through graft versus tumor (GVT) effects.

Fusariosis diseminada por Fusarium oxysporum en un paciente adulto con leucemia mieloide aguda y neutropenia severa febril / Disseminated fusariosis by Fusarium oxysporum in an adult patient with acute myeloid leukemia and severe febrile neutropenia

Reportamos el primer caso de fusariosis diseminada en un paciente adulto en Chile, con una neoplasia hematológica y tratamiento quimioterápico, quien evolucionó con neutropenia febril prolongada, refractaria, fuera tratado con un amplio esquema antibacteriano y desarrollara una infección multisistémica, con compromiso cutáneo, sinusal y pulmonar por Fusarium oxysporum. Cursó con refractariedad al tratamiento antifúngico con anfotericina B deoxicolato y caspofungina, utilizados en forma secuencial. El desenlace fatal de este paciente se asoció a la persistencia de la neutropenia y a la infección por un hongo filamentoso habitualmente resistente a terapia antifúngica.

Autotrasplante (AT) de progenitores hematopoyéticos en mieloma múltiple: Experiencia clínica / Autologous transplant (AT) with peripheral-blood stem-cell rescue for multiple myeloma: A clinical experience

Background:Multiple myeloma is rarely curable. Advances in high dose chemotherapy and stem cell transplantation have improved overall survival and event-free disease periods, but relapses are inevitable. Aim: To report our experience with AT in multiple myeloma, between 1994 and 2003. Material and Methods: Retrospective analysis of 20 patients (12 women), with a mean age of 51.1 years. VAD (vincristine, doxorubicin and dexamethasone) was used as initial therapy in 19 patients. High dose cyclophosphamide (11 patients) and variations of VAD regimen (7) associated with granulocyte colony stimulating factor were used for peripheral-blood stem cell harvest. The conditioning regimen consisted of melphalan 200 mg/m2 followed by the reinfusion of peripheral-blood stem cells 24 hours later. The median number of CD34 cells infused was 3,3x106/kg. Three patients were subjected to a second auto graft and one to a non-myeloablative transplant. Mean follow up was 35.5 months. Results: Mucositis and febrile neutropenia were common complications. The median number of days for neutrophyl engraftment was 9 (range 8-11) and for platelets, 10 (range 7-13). No patient died. Complete remission was obtained in 60% (12/20), progession-free survival was 30 months and overall median survival, 47 months. Conclusions: The AT with high-dose melphalan is a safe procedure in our hospital, without mortality and engraftment in all the patients. Complete remission and progression free survival were similar to those reported abroad but the overall median survival was lower.

Linfoma intravascular tratado con anticuerpos monoclonales anti CD20: descripción de un caso clínico / Intravascular lymphoma treated with anti CD20 monoclonal antibodies: report of one case

We report a 78 year old male with prostatism, that was subjected to a prostate biopsy. The pathological study showed a microvascular lymphocytic infiltration. Four months later, the patients presentd with reduced alertness, cough, dyspnea, fever and elevation of lactic dehydrogenase and erythrocyte sedimentation rate. Chest and abdominal CAT scans, bone marrow aspirate, protein electrophoresis and prostate specific antigen were normal. A re-evaluation of prostate biopsy showed an intravascular lymphoid infiltration, positive for CD45 and CD20, compatible with the diagnosis of intravascular lymphoma. Chemotherapy was started, but it was not tolerated by the patient and the response was partial. Therefore, treatment with monoclonal antibodies anti CD20 (Rituximab) was started. The tumor had a complete and prolonged (24 months) remission after the treatment.

Intoxicación por rodendicida - superwarfarina: descripción de un caso clínico / Poisoning by rodenticide - superwarfarin: description of a clinical case

Los rodendicidas de uso doméstico están fácilmente disponibles en el comercio y, corresponden en general a superwarfarinas, cuyo mecanismo de acción es semejantea los anticoagulantes de uso clínico, pero difiere en una vida media más prolongada (meses) y una potencia cien veces mayor.Presentamos el caso clínico de una intoxicación con fines suicidas de Rodilon® (difethialone) 10 cajas de 50 g. La paciente ingresa con un tiempo de protrombina(TP) de 6 por ciento con un INR de 12.4, requiriendo administración de vitamina K1 durante varios meses. Las manifestaciones clínicas fueron leves (gingivorragia y petequias) al ingreso.En caso de no contar con el antecedente de ingesta, la intoxicación debe sospecharse en toda hipoprotrombinemia adquirida, sin antecedentes familiares de diátesis hemorrágica y, con función hepática normal.