Lymphocyte subsets and soluble forms of MIC-A and MIC-B are prognostic factors in non-Hodgkin lymphoma patients.

MIC-A and MIC-B are the natural ligands for NKG2D, an activator receptor expressed in NK cells. Soluble isoforms of MIC-A and MIC-B (sMICA, sMICB) have been identified in different malignancies, affecting NK cells' cytotoxicity. The study was performed to determine the levels of sMICA, sMICB, the expression of MIC-A, and MIC-B on tumor tissues, and lymphocyte subpopulations (CD4 + , CD8 + , NK, NKT, Tγδ cells, B cells, monocytes) in 94 patients with non-Hodgkin's lymphoma (NHL) and 72 healthy donors.The most frequent lymphoma was diffuse large B cell lymphoma (48%). Patients with NHL had decreased numbers of CD4 T cells, CD8 T cells, B cells, monocytes, NK cells, type 1 dendritic cells, γδ T cells, and increased iNKT cells. Patients showed higher levels of sMIC-A and similar serum levels of sMIC-B.Survival was poorer in patients having higher LDH values and lower numbers of CD4 T cells, type 1 dendritic cells, gamma-delta T cells, and high levels of sMIC-A.In conclusion, high levels of sMIC and decreased numbers in circulating lymphocyte subsets are related to poor outcomes in NHL.

Soluble MIC-A, IPI, and response to treatment strongly predict survival in patients with germinal center diffuse large B cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most frequent lymphoma. MIC-A and MIC-B are the natural ligands for NKG2D, a receptor expressed in NK cells. MIC-A soluble isoforms (sMICA) have been described in different malignancies. OBJECTIVES: To analyze lymphocyte subsets and sMIC-A in germinal center DLBCL. MATERIALS AND METHODS: sMICA, sMICB, and peripheral blood lymphocyte subsets (CD4+, CD8+, NK, NKT, γδ T cells, and dendritic cells) were analyzed in 59 patients and 60 healthy donors. RESULTS: Patients had decreased numbers of type 1 and type 2 dendritic cells, NK, iNKT, CD4 T, and CD8 T cells, and higher levels of sMIC-A. The 2-year PFS for high IPI scores and high sMIC-A was 24% and 28%, respectively. The 2-year OS for high IPI scores and high sMIC-A was 42% and 33%. The 2-year PFS and OS for patients not achieving response to treatment were 0% and 10%, respectively. The MICPI score (one point each for high IPI score and high sMIC-A) showed that those patients summing two points had worse PSF and OS. CONCLUSIONS: Patients with DLBCL have decreased numbers of peripheral lymphocyte subsets and high levels of sMIC-A. The addition of sMIC-A to IPI could improve its prognostic relevance.

Poor Survival in COVID-19 Associated with Lymphopenia and Higher Neutrophile-Lymphocyte Ratio.

BACKGROUND: Immune cell counts in blood in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection may be useful prognostic biomarkers of disease severity, mortality, and response to treatment. OBJECTIVES: To analyze sub-populations of lymphocytes at hospital admission in survivors and deceased from severe pneumonia due to coronavirus disease-2019 (COVID-19). METHODS: We conducted a cross-sectional study of healthcare workers confirmed with SARS-CoV-2 in convalescents (control group) and healthy controls (HC) diagnosed with severe COVID-19. Serum samples were taken at hospital admission and after recovery. Serum samples ≥ 25 days after onset of symptoms were analyzed for lymphocyte subpopulations through flow cytometry. Descriptive statistics, Kruskall-Wallis test, receiver operating characteristic curve, calculation of sensitivity, specificity, predictive values, and Kaplan-Meier analysis were performed. RESULTS: We included 337 patients: 120 HC, 127 convalescents, and 90 severe COVID-19 disease patients (50 survivors, 40 deceased). For T cells, total lymphocytes ≥ 800/µL, CD3+ ≥ 400/µL, CD4+ ≥ 180/µL, CD8+ ≥ 150/µL, B cells CD19+ ≥ 80/µL, and NK ≥ 34/µL subsets were associated with survival in severe COVID-19 disease patients. All subtypes of lymphocytes had higher concentrations in survivors than deceased, but similar between HC and convalescents. Leukocytes ≥ 10.150/µL or neutrophils ≥ 10,000/µL were associated with increased mortality. The neutrophil-to-lymphocyte ratio (NLR) ≥ 8.5 increased the probability of death in severe COVID-19 (odds ratio 11.68). CONCLUSIONS: Total lymphocytes; NLR; and levels of CD3+, CD4+, CD8+, and NK cells are useful as biomarkers of survival or mortality in severe COVID-19 disease and commonly reach normal levels in convalescents.

Assessment and potential-site determination of a wheat straw power plant by Aspen Plus and multi-criteria GIS model.

Wheat is one of the most important crops worldwide. Mexicali, Baja California, is an important wheat producer in Mexico with an average production of 507,543 t. Wheat straw is generated as a residue which could be used for different purposes such as bioenergy, heat and power generation. In this work, an assessment and potential site determination of a biomass power plant operating with wheat straw as fuel was performed. Aspen Plus was used to evaluate a plant capacity of at least 10 MW considering the physicochemical properties and an higher heating value of 14.86 MJ kg-1 of the wheat straw from the region. The combustion produced 39.76 MW, and the overall plant efficiency was 25.52%. The development of the multi-criteria geographic information system model allowed us to assess and analyse four factors and three restrictions to determine the potential site for the biomass power plant. The factors were raw material, wheat crops, electric transmission lines, paths and roads, water canals and aqueducts, while the restrictions were localities, Ramsar sites and faults. The biomass power plant is technically and geographically feasible. The geographical coordinates of the potential site of the biomass power plant that fulfils all the criteria are 32°29'29.72″N and 115°15'39.45″W.

Phenotypes and concentration of PON1 in cardiovascular disease: The role of nutrient intake.

BACKGROUND AND AIMS: Paraoxonase 1 (PON1) is considered to play a crucial role as an anti-atherosclerotic factor. The PON1 activity is affected by genetic polymorphisms, environmental factors, age, sex, lifestyle, pharmaceutical drugs, and dietary factors. The aim of this study was to evaluate the association between macro- and micronutrients as well as PON1 concentration and activities in patients with cardiovascular diseases (CVD), cardiovascular risk factors but no CVD (CRF), and in healthy controls (control group). METHODS AND RESULTS: A case-control study was carried out with 356 volunteers from the Mexican Institute of Social Security, Mexico. Clinical parameters, lipid profile, PON1 activities (AREase, LACase, CMPAase and PONase), and PON1 concentration were evaluated. There was a differential intake of macro- and micronutrients among the study groups. The intake of proteins and carbohydrates was higher in the CVD group than in the CFR and control groups (p < 0.05). AREase, LACase, and CMPAase activities and PON1 concentration were lowest in the CVD group. CONCLUSION: LACase and CMPAase activities, as well as PON1 concentration, could be included in the battery of CVD predictive biomarkers in the Mexican population.

Pathophysiology of Alloimmunization.

INTRODUCTION: Alloimmunization is caused by exposure to erythrocytes from a donor that expresses blood group antigens other than those of the recipient and is related to processes that alter the balance of the immune system. Knowing the pathophysiology of alloimmunization process is essential to understand clinical complications associated with this process. PATIENTS AND METHODS: From October 2016 to April 2017, irregular antibody screening was performed in 1,434 polytransfused (compatible with the ABO and D system) patients by means of agglutination techniques using erythrocytes of a known phenotype of 44 patients with a positive alloantibody screening. Non-alloimmunized (control) subjects were matched for age, gender, pathology, and treatment group with alloimmunized patients. The subsets of B, T, and Treg lymphocytes were determined by flow cytometry. RESULTS: The results of screening for alloantibodies in patients by specificity of antibodies were as follows: nonspecific (30%), followed by anti-Dia (13%), anti-e (9%), anti-S (9%), anti-I (7%), anti-K (7%), and anti-P (7%). A lower percentage of CD4+ T lymphocytes and an increase of CD8+ T lymphocytes were observed in alloimmunized patients, as well as a low CD4/CD8 ratio (0.7 vs. 1.6, p = 0.003), a higher percentage of B lymphocytes versus the control group (30 vs. 20%, p = 0.003), and a decrease of Treg CD4+ lymphocytes versus the control group (3 vs. 12 cells/µL, p = 0.043). These observations suggest that alloimmunized patients have important alterations in the number of some lymphocyte subsets that can be translated into clinical immune dysregulation. CONCLUSION: A decreased CD4/CD8 ratio, increased B lymphocytes, and Treg lymphocyte deficiency are the most significant changes observed in alloimmunized patients.

Plasmid pSfr64a and the symbiotic plasmid pSfr64b of Sinorhizobium fredii GR64 control each other's conjugative transfer through quorum-sensing elements.

Rhizobia are nitrogen-fixing symbionts of plants. Their genomes frequently contain large plasmids, some of which are able to perform conjugative transfer. Plasmid pSfr64a from Sinorhizobium fredii GR64 is a conjugative plasmid, whose transfer is regulated by quorum sensing genes encoded by itself (traR64a, traI64a), in the symbiotic plasmid pSfr64b (traR64b, traI64b), and in the chromosome (ngrI). Also, transfer of pSfr64b requires quorum sensing elements encoded in this plasmid (traR64b, traI64b), in pSfr64a (traR64a), and in the chromosome (ngrI). These results demonstrate that pSfr64a and the symbiotic plasmid depend on each other for conjugative transfer. Plasmid pSfr64a from S. fredii GR64 is unable to transfer from the genomic background of Rhizobium etli CFN42. Our results show that the relaxase of pRet42a is able to process the oriT of pSfr64a, and viceversa, underlining their functional similarity and suggesting that in addition to the external signals, the "cytoplasmic environment" may pose a barrier to plasmid dissemination, even if the plasmids are functional in other aspects.

Maternal obesity associated with increase in natural killer T cells and CD8+ regulatory T cells in cord blood units.

BACKGROUND: One of the major limitations of umbilical cord blood (UCB) as hematopoietic stem cell source is its restricted cell number. In mothers who are candidates for stem cell donation, there are variables that affect the quantity and quality of UCB units. The aim of this study was to determine if obstetric, maternal, and fetal factors modify the number of lymphocyte subsets in UCB units. STUDY DESIGN AND METHODS: This was a prospective, observational study. In UCB units, the numbers of CD34, NK, NKT, iNKT, Type 1 dendritic cells (DCs), Type 2 DCs, T γδ, T CD4+, T CD8+ lymphocytes, CD4+CD25+FoxP3+, and CD8+CD25+FoxP3+ T regulatory (Treg) cells were quantified by flow cytometry. RESULTS: Fifty-four UCB units were included; the donors' mean weight was 75 kg (range, 52 to 102 kg) and they had a mean body mass index (BMI) of 30 kg/m(2) (range 22 to 40 kg/m(2) ), of which 12 (22%) had a normal BMI, 14 (26%) were overweight, and 28 (52%) were obese. The mean number of CD34+ cells was 4.45 × 10(6) (range, 0.7 × 10(6) to 20.5 × 10(6) ). The number of NKT, CD3+, CD4+, CD8+, and CD8+CD25+FoxP3+ Treg cells was significantly higher in overweight or obese mothers; CD34+ cells were decreased in the same group. The number of iNKT and CD34+ cells was decreased in newborns weighing above the average. CONCLUSIONS: Maternal factors such as BMI, and fetal factors such as weight at birth, should be added to the selection criteria of UCB donors.

Perceived environment in relation to objective and self-reported physical activity in Spanish youth. The UP&DOWN study.

The aims of the present study were to assess the association of environmental perception with objective and self-reported physical activity (PA) and the relation between environmental perception and meeting PA recommendations on children and adolescents. A sample of 1520 youth (770 boys) aged 8-18 years (12.1 ± 2.5 years) from the UP&DOWN study were included in the data analyses. Environmental perception was assessed with the short adapted version of the ALPHA environmental questionnaire. PA was objectively (accelerometers) and self-reported measured (PA Questionnaire for Children, Patient-centered Assessment and Counseling for Exercise Questionnaire and Finnish PA Index). Linear regression models were used to assess the association of environmental perception with PA. Bivariate logistic regression models were used to assess differences between environmental perception and meeting PA recommendations. Environmental perception was positively associated with both objective and self-reported PA. Some differences were found in the association of environmental perception and PA between sex- and age-specific groups. Youth who perceived a more favourable environment were more likely to meet PA recommendations (at least 60 min · day(-1) of moderate to vigorous PA (MVPA)). Results suggest that environmental perceptions of children and adolescents may play an important role in achieving higher levels of PA.

Human papillomavirus infections in Mexican women with normal cytology, precancerous lesions, and cervical cancer: type-specific prevalence and HPV coinfections.

The prevalence and genotype distribution of human papillomavirus (HPV) provides the basis for designing HPV prevention programs. The prevalence rates of type-specific HPV and coinfections in samples of Mexican women were investigated in 822 women aged 18-87 years. HPV detection was performed using a Linear Array™ genotyping test. HPV infection was found in 12.4% of controls, 46.3% of those with cervical intraepithelial neoplasia 1, and 100% of those with cervical intraepithelial neoplasia 3 or cervical cancer. HPV 16 was the most prevalent type in all diagnosis groups. The HPV types most frequently found in cervical cancers were 16, 18, 45, 52, 58, and 39; HPV types 16, 62, 51, 84, 18, 53, and CP6108 were the most prevalent in control women. Considering HPV-positive samples only, coinfections occurred most often in controls (63%) and were less frequent in those with cervical cancer (26%). The most frequent viral types in coinfections with HPV 16 in control women were HPV 62, 51, and 84; in women with cervical cancers, HPV 18, 39, and 70 were most common. In conclusion, in addition to HPV types 16 and 18, types 45, 39, 58, 52, and 71 were found in cervical cancers in Mexican women (78%); among them, only 65% were attributable to HPV types 16 and 18. Therefore, it is necessary to consider these viral types in the design of new vaccines, and to determine whether certain HPV types coinfecting with HPV 16 in precursor lesions determine tumor progression or regression.

Morbidity of breast cancer and cervico-uterine cancer in women from the occidental region of Mexico.

BACKGROUND: The incidences of breast cancer (BC) and cervico-uterine cancer (CC) vary widely from country to country. In Mexico, BC mortality has doubled in the last 20 years to become the second leading cause of death for women aged 30 to 54 years. CC is the most common cause of death from neoplasia in women over 25 years old. In 2006, the state of Nayarit had one of the highest mortality rates for these types of cancers in Mexico. OBJECTIVE: To analyze and characterize the current demographics and morbidities associated with BC and CC in the state of Nayarit. MATERIAL AND METHODS: In this retrospective study, the clinical histories of patients who were diagnosed with BC or CC at the State Cancer Center from January 2006 to December 2010 were analyzed. RESULTS: A total of 406 patients with BC and 328 patients with CC were registered. The most common clinical stage for both cancer types was IIB. The municipalities of San Pedro Lagunillas and El Nayar presented the highest prevalences of BC and CC, respectively. CONCLUISION: Our results suggest that women living in poorer and more marginalized regions have a higher possibility of developing BC and CC. Because BC and CC are preventable and treatable in their early stages, demographic information from population records for these cancers is helpful in determining the incidence rates and patterns and improving decision-making processes.

Significant increase in MIC-A and MIC-B and soluble MIC-A and MIC-B in canine lymphomas.

Non-Hodkin's lymphoma (NHL) is the most frequent hematologic malignancy in humans and dogs. NKG2D is one of the most critical receptors on NK cells, recognizing their natural ligands on malignant cells such as A and B major histocompatibility complex-related proteins (MIC-A and MIC-B). Soluble molecules (sMIC-A and sMIC-B) can interfere with immune synapsis between NK cells and tumor cells, impeding NK cytotoxicity. The main objectives of this study were to analyze, in dogs with diffuse large B cell lymphoma, NK cell lymphoma, and reactive lymphadenopathies, the role of NK cells, their activating receptors NKG2D and NKp46, and their ligands MIC-A and MIC-B, as well as soluble molecules sMIC-A and sMIC-B. Thirty-six dogs with a possible diagnosis of NHL and eight healthy dogs were studied. NHL was diagnosed in 28 (78 %) dogs; in the other 8 (22 %), reactive lymphadenopathies were present. Most of the lymphomas corresponded to B cell NHL (82 %). The most predominant subtype was diffuse large B cell lymphoma (21, 71.5 %), followed by five cases (18 %) that were Non-B Non-T lymphomas (presumably NK cell lymphomas) and other B cell lymphomas (3, 10.5%). There were no cases of T cell NHL. MIC-A was positive in 7 of 27 (26 %) cases of NHL, and MIC-B in 20 of 27 (74 %) NHL. In non-malignant lymphadenopathies, three (37.5 %) dogs were positive for MIC-A, and five (62.5 %) expressed MIC-B. Dogs with lymphoma had higher numbers of NK cells than eight healthy dogs. In 15 dogs (12 cases with NHL and three cases with reactive adenopathies) and eight controls, there were no differences in the number of NK cells expressing NKP46 and NKG2D. NHL dogs had higher values of sMIC-A and sMIC-B. B-cell and NK cell lymphomas correspond to 86 % and 14 % of all canine lymphomas. MIC-A, MIC-B, and sMIC-A and sMIC-B were increased in canine lymphomas.

[Impact of hyperglycemia on the prognosis of patients hospitalized for severe pneumonia in COVID-19]. / Impacto de hiperglucemia en el pronóstico de hospitalizados por neumonía grave en COVID-19.

Background: Adequate glycemic control improves the prognosis of patients hospitalized for pneumonia associated with severe COVID-19. Objective: To evaluate the impact of hyperglycemia (HG) on the prognosis of patients hospitalized for severe pneumonia associated with COVID-19 in unvaccinated patients. Material and methods: Prospective cohort study. We included patients hospitalized from August 2020 to February 2021, with severe COVID-19 pneumonia, not vaccinated against SARS-CoV-2. Data was collected from admission to discharge. We used descriptive and analytical statistics according to the data distribution. ROC curves were used to determine the cut-off points with the highest predictive performance for HG and mortality, with the IBM SPSS program, version 25. Results: We included 103 patients, 32% women, 68% men, age 57 ± 13 years; 58% were admitted with HG (191, IQR 152-300 mg/dL) and 42% with normoglycemia (NG < 126 mg/dL). Mortality was higher in HG at admission 34 (56.7%) than in NG 13 (30.2%) (p = 0.008). HG was associated with diabetes mellitus 2 and neutrophilia (p < 0.05). The risk of death increases 1.558 times (95% CI 1.118-2.172) if HG is at admission and 1.43 times (95% CI 1.14-1.79) during hospitalization. Maintaining NG throughout the hospitalization contributed independently to survival (RR = 0.083 [95% CI 0.012-0.571], p = 0.011). Conclusion: HG significantly impacts prognosis by increasing mortality more than 50% during hospitalization for COVID-19.

Introducción: el adecuado control glucémico mejora el pronóstico de pacientes hospitalizados por neumonía asociada a COVID-19 grave. Objetivo: evaluar el impacto de la hiperglucemia (HG) sobre el pronóstico de pacientes hospitalizados por neumonía grave asociada a COVID-19 en no vacunados. Material y métodos: estudio de cohorte prospectivo. Se incluyeron pacientes hospitalizados de agosto de 2020 a febrero de 2021, con neumonía grave por COVID-19, no vacunados contra SARS-CoV-2. Los datos fueron recolectados desde el ingreso hasta el egreso. Se empleó estadística descriptiva y analítica de acuerdo con la distribución de datos. Se construyeron curvas ROC para determinar los puntos de corte de mayor rendimiento predictivo para HG y mortalidad, con el programa IBM SPSS, versión 25. Resultados: se incluyeron 103 pacientes, 32% mujeres, 68% hombres, edad 57 ± 13 años; 58% ingresaron con HG (191, IQR 152-300 mg/dL) y 42% en normoglucemia (NG < 126 mg/dL). La mortalidad fue mayor en HG al ingreso 34 (56.7%) que en NG 13 (30.2%) (p = 0.008). La HG se asoció con diabetes mellitus 2 y neutrofilia (p < 0.05). El riesgo de muerte se incrementó 1.558 veces (IC 95% 1.118-2.172) si la HG fue al ingreso y 1.43 veces (IC 95% 1.14-1.79) durante la hospitalización. Mantener NG durante todo el internamiento contribuyó de manera independiente a la sobrevida (RR 0.083 [IC 95% 0.012-0.571], p = 0.011). Conclusión: la HG impacta significativamente el pronóstico al incrementar en más de 50% la mortalidad durante la hospitalización por COVID-19.

CD133+CD34+ and CD133+CD38+ blood progenitor cells as predictors of platelet engraftment in patients undergoing autologous peripheral blood stem cell transplantation.

BACKGROUND: Hematopoietic stem cells (HSC) have been characterized by CD34+ expression and an adequate dose of CD34+ cells is associated with a complete engraftment. CD133 is a more specific marker of HSC. MATERIALS AND METHODS: We studied the relationship between graft content of CD34+, CD133+, and CD38+ cells and trilineage engraftment after autologous stem cell transplantation in patients with different hematological disorders. Blood samples were obtained before and after mobilization with recombinant granulocyte-colony stimulating factor (G-CSF, 16 µg/kg), from apheresis collections, and after transplantation. RESULTS: Cell subsets were quantified by flow cytometry, and the dose of each population infused was correlated with success of engraftment. G-CSF induced mobilization of CD133+CD38+ cells (12.6-fold) and CD133+CD34+ cells (14.7-fold). A correlation was observed between the infused dose of CD133+CD34+ and CD133+CD38+ cells and platelet engraftment. CONCLUSION: CD133+CD34+ and CD133+CD38+ cells were mobilized with G-CSF and these cell subsets were correlated with platelet engraftment.

Undifferentiated immunophenotypes and not expression of BCR-ABL can be associated in adult Mestizo Mexican patients with ALL.

INTRODUCTION: The BCR-ABL t(9;22)(q34;q11) translocation has been identified as a risk factor in de novo acute lymphoblastic leukemia (ALL), but there are other factors that may influence survival in patients not expressing this translocation. OBJECTIVE: To associate expression and non-expression of BCR-ABL with immunophenotype and other clinical features in adult patients with ALL from a Mexican mestizo population. MATERIAL AND METHODS; Peripheral blood samples from 35 adult patients with de novo ALL were used to detect BCR-ABL by reverse transcriptase polymerase chain reaction (RT-PCR) as well as immunophenotype by flow cytometry. RESULTS: In the group of BCR-ABL negative patients (74.28%) two subgroups were identified with the immature immunophenotypes CD34+/CD33+ and/or CD13+, and CD10-/CD34+. In the group of BCR-ABL positive patients (25.72%) leukemic blast cells with a more differentiated immunophenotype compared to the BCR-ABL negative group were found. As regards clinical and biological characteristics, we found survival in months to be very similar and a tendency to high initial leukocyte counts in both groups. CONCLUSIONS: This is the first study conducted on a Mexican mestizo population to report that BCR-ABL negative patients can present a high frequency of undifferentiated immunophenotypes and must therefore be considered as vulnerable as BCR-ABL positive patients.

Serum calcium is an independent prognostic factor of overall survival in Mexican patients with multiple myeloma.

OBJECTIVE: To evaluate the impact of different prognostic factors that has been suggested to be useful in predicting the survival of patients with multiple myeloma (MM). MATERIALS AND METHODS: A longitudinal prospective study was conducted on 24 adult Mexican patients diagnosed with primary MM. The levels of expression of CD38, CD138 and cyclin D1 were analyzed in plasma cells (PCs) from patients and mononuclear cells from healthy donors. Serum levels of lactate dehydrogenase, creatinine, calcium, beta2 microglobulin and interleukin-6 (IL-6) as well as hemoglobin and platelet count were taken into consideration. RESULTS; CD138 and cyclin D1 levels in absolute numbers were significantly overexpressed in malignant PCs. A positive correlation was noted between cyclin D1 and CD38 expression levels in malignant PCs. IL-6 and serum calcium were also positively correlated in MM patients. Cyclin D1 overexpression was not associated with better overall survival (OS). Normal calcium levels were associated with better overall survival (OS). Serum calcium was the only variable correlating with better OS in Cox regression analysis. CONCLUSION: Serum calcium is an independent prognostic factor of OS in a population of Mexican patients with MM.

The conjugative plasmid of a bean-nodulating Sinorhizobium fredii strain is assembled from sequences of two Rhizobium plasmids and the chromosome of a Sinorhizobium strain.

BACKGROUND: Bean-nodulating Rhizobium etli originated in Mesoamerica, while soybean-nodulating Sinorhizobium fredii evolved in East Asia. S. fredii strains, such as GR64, have been isolated from bean nodules in Spain, suggesting the occurrence of conjugative transfer events between introduced and native strains. In R. etli CFN42, transfer of the symbiotic plasmid (pRet42d) requires cointegration with the endogenous self-transmissible plasmid pRet42a. Aiming at further understanding the generation of diversity among bean nodulating strains, we analyzed the plasmids of S. fredii GR64: pSfr64a and pSfr64b (symbiotic plasmid). RESULTS: The conjugative transfer of the plasmids of strain GR64 was analyzed. Plasmid pSfr64a was self-transmissible, and required for transfer of the symbiotic plasmid. We sequenced pSfr64a, finding 166 ORFs. pSfr64a showed three large segments of different evolutionary origins; the first one presented 38 ORFs that were highly similar to genes located on the chromosome of Sinorhizobium strain NGR234; the second one harbored 51 ORFs with highest similarity to genes from pRet42d, including the replication, but not the symbiosis genes. Accordingly, pSfr64a was incompatible with the R. etli CFN42 symbiotic plasmid, but did not contribute to symbiosis. The third segment contained 36 ORFs with highest similarity to genes localized on pRet42a, 20 of them involved in conjugative transfer. Plasmid pRet42a was unable to substitute pSfr64a for induction of pSym transfer, and its own transfer was significantly diminished in GR64 background. The symbiotic plasmid pSfr64b was found to differ from typical R. etli symbiotic plasmids. CONCLUSIONS: S. fredii GR64 contains a chimeric transmissible plasmid, with segments from two R. etli plasmids and a S. fredii chromosome, and a symbiotic plasmid different from the one usually found in R. etli bv phaseoli. We infer that these plasmids originated through the transfer of a symbiotic-conjugative-plasmid cointegrate from R. etli to a S. fredii strain, and at least two recombination events among the R. etli plasmids and the S. fredii genome. As in R. etli CFN42, the S. fredii GR64 transmissible plasmid is required for the conjugative transfer of the symbiotic plasmid. In spite of the similarity in the conjugation related genes, the transfer process of these plasmids shows a host-specific behaviour.

FLT3-ITD and CD135 Over-Expression are Frequent Findings of Poor Survival in Adult Patients with Acute Leukemias.

BACKGROUND: Fms-like tyrosine kinase 3 (FLT3) expression and mutation have been considered a poor prognostic factor in acute myeloid leukemia (AML). FLT3-ITD mutation is present in 30% of adult patients with AML and 2-5% in childhood acute lymphoblastic leukemia (ALL). The impact of these mutations on the prognosis of ALL patients, has not yet been established. Moreover, a limited number of publications regarding the level of expression of the FLT3 receptor (CD135) in both leukemias exist. This study aimed to analyze the clinical outcomes associated to the presence of FLT3-ITD mutation and the expression of CD135. METHODS: 82 adult patients with newly diagnosed acute leukemia (39 with AML and 43 with ALL) were included. Flow cytometry and RT-PCR were done to analyze the expression of CD135 and the presence of FLT3 ITD mutation, respectively. RESULTS: FLT3-ITD was present in 14 (36%) of AML and 15 (35%) of ALL patients. Disease free survival (DFS) and overall survival (OS) were lower in ALL patients having a CD135 expression >3000 cells/µL. There was a trend for poor OS in AML patients expressing FLT3 ITD. OS was worse in AML patients with high expression of CD135. CONCLUSION: A higher (35%) frequency of FLT3-ITD was found in adult ALL patients. The presence of FLT3-ITD was associated with a trend of poor OS in AML cases, and overexpression of CD135 was correlated with poor DFS in ALL cases and poor OS in both acute leukemias.

Important decrease in invariant natural killer T, CD4+ regulatory T cells, CD8+ regulatory T cells, gamma-delta T cells, and CD4+ T lymphocytes in HIV-negative patients with hemophilia.

Hereditary hemophilias are X-linked inherited bleeding disorders defined as deficiencies of the coagulation factors VIII or IX. They are characterized by easy to provoke or spontaneous bleeding. HIV infection in hemophilic patients is a risk factor for the reduction of CD4+ T cells. There is no information regarding the cellular immune function in HIV-negative patients with hemophilia. To evaluate the number of lymphocyte subsets in adult patients with hemophilia A or B as compared with healthy donors. 39 Adult hemophilics and 27 healthy donors were included. Lymphocyte subsets [CD4 and CD8 T cells, natural killer cells, natural killer T (NKT) cells, invariant NKT (iNKT) cells, gamma-delta T (γδT) cells, type 1 and 2 dendritic cells, CD14 monocytes, CD4 and CD8 regulatory T cells (Tregs), and B cells], were analyzed by flow cytometry. A significant decrease of CD4+ T lymphocytes, γδT cells, iNKT cells, CD4+ and CD8+ Tregs was observed in patients with hemophilia. Those patients having factor VIII inhibitor had the lowest CD4+ Treg and CD8+ Treg counts. CD14 monocytes were increased, as well as iNKT and type 2 dendritic cells in obese-overweight hemophilics. CD4+ lymphocytes, iNKT, γδT cells, and Tregs (CD4+ and CD8+), are significantly decreased in patients with hemophilia. Depletion of Tregs is more important in patients with factor VIII inhibitor. Physicians caring for hemophilia patients should realize that, even when they are not suffering infections frequently, may have early evidence of cellular immunodeficiency.

Clinical relevance of NKT cells and soluble MIC-A in Hodgkin lymphoma.

Previous studies demonstrated that the majority of Hodgkin lymphoma (HL) patients achieve response after treatment, while 5% become refractory. Studies analyzing the role of lymphocyte subsets in peripheral blood are limited. This investigation sought to evaluate peripheral blood lymphocyte subsets and soluble MHC class I chain-related proteins A and B (sMIC-A/B) and their correlation with survival in patients with newly diagnosed HL. The study recruited 36 patients and 72 healthy donors. HL patients showed a decrease in CD4, B, monocytes, NK, and NKT cells; and an increase in γ-δ T cells and soluble MIC-A serum levels. Higher values of s-MIC-A  >100 ng/mL and NKT cells >40 µL correlated with poor overall survival (OS). In conclusion, in HL peripheral blood CD4 T and B cells, monocytes, NK, and NKT cells were decreased, while s-MIC-A and γ-δ T cells increased. Higher values of s-MIC-A and NKT cells correlated with poor survival.