Response of early-cycle common bean cultivars with carioca, black and speckled grains to top-dressing nitrogen fertilization

The aim was to evaluate the agronomic and qualitative attributes of early-cycle common bean cultivars with different grains types grains in response to top-dressing nitrogen (N) doses. The experiment was carried out using a randomized block design, in a split-plot scheme, with 4 replicates. The plots consisted of the cultivars IAC Nuance, IAC 1849 Polaco and IAC Veloz, with speckled, Carioca and black grains, respectively. The subplots were formed by N doses applied as top-dressing: 0 kg ha-1, 60 kg ha-1 (applied in the stage of third trifoliate leaf), 120 kg ha-1 (1/2 applied at third trifoliate leaf stage + 1/2 applied at the floral bud stage) and 180 kg ha-1 (1/3 applied at the first trifoliate leaf stage + 1/3 applied at the third trifoliate leaf stage + 1/3 applied at the floral bud stage). IAC Veloz stood out for grain yield, showing the highest grain yield in the lowest N doses, being classified as efficient to the use of N. The cultivars IAC Nuance and IAC 1849 Polaco reached maximum yields with 155 and 163 kg ha-1 of N. The IAC Nuance was the most responsive, increasing grain yield by up to 25.3% due to nitrogen fertilization. Increasing N doses applied as top-dressing increased the sieve yield and crude protein content of the common bean cultivars, with IAC Nuance standing out. The cultivars showed good grain quality, and IAC 1849 Polaco and IAC Veloz had the shortest cooking time and IAC Veloz also had the fastest hydration.

Tableta masticable de lecitina de soya: de subproducto a producto farmacéutico / Chewable pill of soy phosphatidylcholine: from by-product to pharmaceutical product

En el Laboratorio Farmacéutico Oriente de Santiago de Cuba se acometió el desarrollo de una tableta masticable de lecitina de soya con fines de registro y ulterior producción, lo cual se realizó durante el bienio 2011-2013. Se utilizaron excipientes de calidad farmacéutica, los métodos analíticos de la Farmacopea de los Estados Unidos, edición 35/Formulario Nacional, edición 30 del 2012, así como la tecnología de granulación húmeda y compresión directa. La lecitina fue caracterizada como materia prima farmacéutica y la tableta desarrollada cumplió con los atributos de calidad establecidos, por lo cual se registró con estabilidad comprobada de 2 años. Se suministró valor agregado a esta sustancia, con riesgo potencial de acumulación para el medio ambiente, como producto farmacéutico nuevo en Cuba

The development of a chewable pill of soy phosphatidylcholilne was undertaken in Oriente Pharmaceutical Laboratory from Santiago de Cuba with registration ends and subsequent production, that was carried out during the biennium 2011-2013. Excipients of pharmaceutical quality, the analytic methods of the United States Pharmacopoeia, edition 35/National Form, 2012 30th edition, as well as the technology of humid granulation and direct compression were used. Phosphatidylcholine was characterized as pharmaceutical raw material and the developed pill fulfilled the established quality attributes, reason why it was registered with 2 years proven stability. Added value was given to this substance, with potential risk of accumulation for the environment, as new pharmaceutical product in Cuba

Aspectos farmacológicos de la lecitina de soya y sus posibles aplicaciones médicas / Pharmacological aspects of soy phosphatidylcholine and its possible medical uses

La lecitina de soya, producto natural empleado como suplemento nutricional, presenta múltiples acciones biológicas demostradas, por lo cual resulta muy beneficiosa para tratar a pacientes con distintas afecciones. Teniendo en cuenta lo anterior se realizó la presente investigación donde se exponen algunos aspectos de interés, con vistas a difundir aún más lo relacionado con esta temática

The soy phosphatidylcholine, natural product used as nutritional supplement, presents multiple demonstrated biological actions, reason why it is very beneficial to treat patients with different disorders. Taking into account the above-mentioned the present investigation was carried out where some aspects of interest are exposed, aimed at diffusing even more everything related to this thematic

Estudio de la capacidad antioxidante in vitro de Phaseolus vulgaris L. (frijol) mediante extracción asistida por microondas / Study of in vitro antioxidant capacity of Phaseolus vulgaris L. (frijol) by microwave assisted extraction

Introducción: Phaseolus vulgaris L. (frijol) es una fuente nutricional importante en Colombia, que aporta un gran contenido de sustancias bioactivas con potencial benéfico para la salud, tales como polifenoles, entre otras, que contribuyen de manera sinérgica con sus propiedades terapéuticas, y que pueden tener un efecto positivo contra algunas patologías. Objetivos: evaluar el método de extracción asistido por microondas como método alternativo para estudiar la capacidad antioxidante in vitro en ocho variedades de P. vulgaris L. cultivadas en Colombia. Métodos: semillas sin piel de P. vulgaris, deshidratadas y maceradas se sometieron a extracción asistida por microondas y extracción sólido-líquido; el contenido de fenoles se evaluó por el método de Folin-Ciocalteu y el potencial antioxidante in vitro se evaluó con base en los métodos del radical estable catión radical difenil-picrilhidrazilo y el radical catión 2,2´-azino-bis(ácido 3-etilbenzotiazolin-6-sulfunico). Resultados: el método de extracción asistido por microondas realizada en horno microondas convencional fue más eficiente respecto a la convencional ya que disminuyó la cantidad de solvente, de muestra empleada y los tiempos de extracción. Los extractos obtenidos por extracción asistida por microondas en horno microndas convencional presentaron un contenido de fenoles entre 29,36 y 60,61 g EAG/L, mientras que el método extracción sólido-líquido, estuvo entre 32,75 y 113,27 g EAG/L. El efecto anti-radicalario fue similar entre los extractos evaluados. Conclusiones: todos los extractos presentaron buena capacidad protectora contra radicales libres, y la técnica de extracción asistida por microondas en horno microndas convencional puede ser usada como método alterno para una valoración rápida, eficiente y eficaz del contenido de sustancias bioactivas en diferentes matrices, se presentó mínimas diferencias entre los resultados obtenidos, comparados con las metodologías de extracción asistida por microondas establecidas antes(AU)

Introduction: Phaseolus vulgaris L. is a representative crop of nutrient and economic importance in Colombia. Additionally, P. vulgaris is considered as a natural source of bioactives compounds, such as polyphenols, which have been associated with valuable effects on health. Objetives: to evaluate the microwave extraction assisted technique as an alternative methodology to study the antioxidant capacity of eight varieties of P. vulgaris cultivated in Colombia. Methods: dehydrated and powered seeds of P. vulgaris was subjected to microwave assisted extraction and solid-liquid extraction and. Total phenolis content was determined by Folin-Cicoulteau method and the potential antioxidant was evaluated using diphenyl-picrylhydrazyl radical stable and 2,2-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical cation assays. Results: microwave assisted extraction-Household microwave oven technique was more efficient and versatile than SLE method. The extracts obtained microwave assisted extraction-Household microwave oven methodology showed polyphenol content ranged between entre 29,36 and 60,61 g EGA/L, but SLE was over 32,75 and 113,27 g EAG/L. Conclusions: all extracts showed a considerable antioxidant potential against free radical, and microwave assisted extraction-Household microwave oven method could be used as an alternative method for fast, efficient and effective evaluation of the content of polyphenol in various matrices, with minimal differences comparing to established microwave assisted extraction techniques(AU)

Isolation and partial characterization of bean phytohemagglutinins

Los extractos de semillas de 21 variedades de caraota fueron ensayados para determinar la especificidad hemaglutinante y la actividad mitogénica. Entre las diferentes variedades de caraotas se pueden distinguir cuatro tipos, dos de los cuales son mitógenos. Se aislaron dos fracciones de lectinas (α y b) de cada uno de los cuatro tipos de caraotas. Sus PM fueron estimados por cromatografía de exclusión y los azúcares presentes por cromatografía en papel. La actividad hemaglutinante, la inhibición de la acción hemaglutinante por derivados de azúcares y glucopéptidos, así como la acción mitogénica, se determinaron para las ocho lectinas purificadas y las cuatro preparaciones control. Las fracciones α y b aisladas a partir de dos de los tipos de caraotas mostraron solamente acción mitogénica mínima, mientras que las de los otros dos tipos de caraotas y todas las preparaciones control fueron mitógenos potentes. Todas las preparaciones mitogénicas aglutinaron en altas diluciones tanto a los glóbulos rojos de vaca activados con tripsina como a los de hámster activados con pronasa; sin embargo, algunas preparaciones resultaron inactivas cuando se probaron con los glóbulos rojos humanos o de conejo(AU)

Extracts of seeds of 21 bean cultlvars were screened for hemagglutinating specifity and for mitogenic activity. Four types could be distinguished in different beans, two of which are mitogens. Two lectin fractions (a and β) were isolated from each of the four bean types. Their MW were estimated by exclusion chromatography and component sugars by paper chromatography. Hemagglutinating activity, inhibition of hemagglutinating action by sugar-derivatives and glyco-peptides as well as mitogenic action were determined for the eight purified lectins and four control preparations. The a and β-fractions isolated from two bean types had only minimal mitogenic action, while those from the other two bean types and all of the control preparations were potent mitogens. All the mitogenic preparations agglutinated trypsin-activated cow red blood cells and pronase-activated hamster red blood cells in high dilutions but some were inactlve when tested with human or rabbit red blood cells(AU)

Expansion and function of MHC restricted killer T cells derived from umbilical cord blood / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to expand the cytotoxic T lymphocytes (CTL) through inducing the differentiation of umbilical blood monomuclear cells (UBMNC) by using various combination of cytokines, and to investigate the functions of expanded CTL.</p><p><b>METHODS</b>The MNC were isolated by ficoll density gradient centrifugation. Then, the PHA-P, IFN-γ combined with IL-2, IL-15 and other cytokines were used for induction and expansion of the cord blood-derived CTL. The biological function of CTL was examined by phenotype analysis, cytotoxic tests and real-time fluorescence quantitative PCR.</p><p><b>RESULTS</b>After expansion for 15 days, the cell number increased by 1522% ± 137%. The content of CD3(-)CD8(-) cells in uncultured cord blood MNC was 95%, and the CD3(+)CD8(+) CTL cells reached 82.77% in cultured cord blood MNC after expansion for 15 days. The expanded CTL cell showed the cytotoxic activity against K562 and HeLa cell line. The killing rate of MNC was 61.88 ± 1.08%. After expansion, the killing rate could reach to 90% with the average value of 90.33 ± 2.02%. The expanded CTL cells highly expressed some key cytokines, such as granzyme A, granzyme B, GM-CSF, granulysin, IFN-γ, TGF-β, TNF-α and perforin. Compared with the control group, the expression of IFN-γ and TGF-β significantly increased (P < 0.05), and the other factors dramatically increased (P < 0.01).</p><p><b>CONCLUSION</b>The cord blood-derived CTL can be expanded by different combinations of cytokines. These protocols may provide alternative choices for CTL cell expansion in tumor adoptive immunotherapy.</p>

Decreased memory T-cell response and function in human immunodeficiency virus-infected patients with tegumentary leishmaniasis

The effects of human immunodeficiency virus (HIV) on the immune response in patients with cutaneous leishmaniasis have not yet been fully delineated. This study quantified and evaluated the function of memory T-cell subsets in response to soluble Leishmania antigens (SLA) from patients coinfected with HIV and Leishmania with tegumentary leishmaniasis (TL). Eight TL/HIV coinfected subjects and 10 HIV seronegative subjects with TL were evaluated. The proliferative response of CD4+and CD8+T-cells and naïve, central memory (CM) and effector memory (EM) CD4+T-cells in response to SLA were quantified using flow cytometry. The median cell division indices for CD4+and CD8+T-cells of coinfected patients in response to SLA were significantly lower than those in patients with Leishmania monoinfection (p < 0.05). The proportions of CM and EM CD4+T-cells in response to SLA were similar between the coinfected patients and patients with Leishmania monoinfection. However, the median CM and EM CD4+T-cell counts from coinfected patients were significantly lower (p < 0.05). The reduction in the lymphoproliferative response to Leishmania antigens coincides with the decrease in the absolute numbers of both EM and CM CD4+T-cells in response to Leishmania antigens in patients coinfected with HIV/Leishmania.

Biological characteristics of PHA-induced CIK cells and its killing activity to K562 cells / 中国实验血液学杂志

The purpose of study was to investigate the in vitro proliferation ability of PHA-induced CIK cells and traditionally prepared CIK cells, the effector cell level and its influence on killing activity to K562 cells, and to analyze the difference between them. The peripheral blood mononuclear cells(PBMNC) of healthy persons were isolated and divided into A and B group. The CIK cells in A group were obtained by using traditional culture method, the CIK cells in B group were prepared by PHA induction. During the cultivation, the cell survival rate and cell absolute value in the cell culture system were counted every 3 days. On day 15 of culture, the cell immunophenotype of 2 groups were detected by flow cytometry, and the ratios of CD3(+)CD56(+), CD3(+)CD8(+) and CD3(+)CD4(+) cells in total cell amount of culture system were accounted. Meantime, the killing activity to K562 cells in different effector-target ratios was detected by using CCK-8 kit between the 2 groups. The results showed that the method of preparing CIK by PHA induction promoted the cell proliferation more than that of the traditional method (P < 0.05), moreover, both the survival rate of cells in 2 groups was more than 90%. The CD3(+)CD8(+), CD3(+)CD56(+) cell ratio in 2 groups obviously increased. As compared with traditional method, the CD3(+)CD8(+) cell level in B group was enhanced (P < 0.05); but there were no statistical differences in increase of CD3(+)CD56(+) cell level and decrease of CD3(+)CD4(+) cell level between 2 groups. while the effector-target ratio is 5:1, 10:1, 20:1 and 40:1, the killing activity of PHA-induced CIK cells to K562 cells was more stronger than traditionally-prepared CIK cells (P < 0.05), moreover, along with increase of effector-target ratio, the difference of killing activity to K562 cells in 2 groups significantly increased. It is concluded that compared with traditional method for preparing CIK cells, the new way by PHA induction can increase the proliferation of CIK cells obviously, enhance the ratio of CD3(+)CD8(+) cells and strengthen the killing activity to the K562 cells. This new way provides a new source of CIK cells and reliable evidence for cyto-immune therapy of leukemia and other tumors.

Alteration of the glycan profile of serum glycoproteins during the seroconversion process in hepatitis B virus-infected patients treated with antiviral therapy and its clinical significance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To use a lectin microarray to study the alteration of glycan affinity profiles of serum glycoproteins during the hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) seroconversion in patients with chronic hepatitis B (CHB) following treatment with antiviral therapy,and to explore its biological significance.</p><p><b>METHODS</b>CHB patients were divided into the following four groups:untreated HBeAg-positive,HBeAg seroconversion after anti-HBV therapy,HBsAg loss after anti-HBV therapy,and healthy individuals (controls).Serum samples were collected from each participant,depleted of high abundance proteins and analyzed by a lectin microarray containing 50 lectins.The lectin-affinity glycan profiles of serum proteins were partially verified by lectin blotting.Between-group differences were analyzed by one-way analysis of variance,and pairwise comparisons were carried out with the Student-Newman-Keuls (SNK) method.</p><p><b>RESULTS</b>The results from the lectin microarray and lectin blotting assay showed significantly reduced affinity for 16 lectins in the untreated HBeAg-positive group compared to the control group (P less than 0.05);in addition,the specific glycan profiles of the untreated HBeAg-positive group included decreased terminal and core fructose,GalNAc alpha-Thr/Ser (T,Tn-antigen),GalNac alpha,terminal beta1-4,and beta-D galactose,bisecting and/or GlcNAc,mannose and Sia.However,the HBeAg seroconversion after anti-HBV therapy group showed enhanced binding of PSA,MPL and the above-mentioned 16 lectins (P less than 0.05),suggesting that the reduced serum glycoprotein glycan structures returned to normal or slightly higher than healthy levels after the therapy-induced seroconversion.Comparison of the group with HBsAg loss after anti-HBV therapy to the group with HBeAg seroconversion after anti-HBV therapy showed the binding ability of ten lectins (AAL,ACL,HAL,HPL,RCA-I,LEL,STL,PHA-E,NML and PCL) were weakened to near control levels and six lectins (VAL,LCA,GNL,PSA,MPL and JAC) were significantly strengthened (all P less than 0.05). These findings implied that the glycan containing terminal fructose, GalNacalpha, terminal beta1-4 galactose,and bisecting GlcNAc glycan structures dropped to near control levels, while the terminal beta-D-galactose residues and core fructose structure increased significantly.</p><p><b>CONCLUSION</b>The glycan structures of serum glycoproteins are closely related to HBeAg and HBsAg seroconversion in CHB patients.It is possible that a special lectin binding glycan involving the terminal beta-D-galactose residues and core fructose may act as sugar markers associated with the disappearance of serum HBsAg during anti-HBV therapy for CHB.</p>

Hydrotime model can describe the response of common bean (Phaseolus vulgaris L) seeds to temperature and reduced water potential / O modelo 'hydrotime' pode descrever a resposta de sementes de feijão comum (Phaseolus vulgaris L) à temperatura e potenciais de água reduzidos

Germination parameters of the response to temperature and water potential from four common bean (Phaseolus vulgaris) lines based on thermal-time and hydrotime concepts were estimated to verify to what extent they can predict germination under different thermal and water conditions. The cultivars IPR Uirapuru and IAPAR 81 (drought-tolerant), and Grauna and Carioca (not tolerant) were used. The isothermal assays were performed in a temperature gradient block, and the assays with different osmotic potentials (PEG 6000) were performed in germination chambers. Seeds from drought-tolerant cultivars spent less time to germinate at supra-optimum temperatures than non-tolerant ones, and the cultivar Uirapuru (drought-tolerant) germinated faster in response to reduced Ψ and low temperatures. The parameter Ψb(50) did not discriminate between drought-tolerant and non-tolerant lines at the infra-optimum temperature range, but it can be used to identify drought-tolerant lines at high temperatures. In general, the hydrotime model reproduced the actual germination data relatively well, chiefly at higher temperatures. This study evidenced that the hydrotime model can be used to describe the germination of common bean seeds under reduced water potentials, and as a screening tool for drought-tolerant bean genotypes.

Neste trabalho foram determinados alguns parâmetros térmicos e hídricos da germinação de quatro cultivares de feijão comum (Phaseolus vulgaris), com base nos modelos graus.dia (thermal-time) e psi.dia (hydrotime), testando-se sua adequação em descrever a germinação em diferentes condições de temperatura e água. Foram analisadas as cultivares IPR Uirapuru e IAPAR 81 (tolerantes à seca), e Grauna e Carioca (não-tolerantes). Os ensaios isotérmicos foram realizados em bloco termogradiente, enquanto que os experimentos com diferentes potenciais osmóticos (PEG 6000) foram realizados em câmaras de germinação. Sementes das cultivares tolerantes à seca germinaram mais rapidamente do que as não-tolerantes em temperaturas supraótimas, sendo que a cultivar Uirapuru (tolerante à seca) germinou mais rápido do que as demais em resposta ao Ψ reduzido e temperaturas baixas. O parâmetro Ψb(50) não discriminou entre cultivares tolerantes e não-tolerantes à seca na faixa térmica infraótima, embora ele possa ser usado para identificar cultivares tolerantes em temperaturas supraótimas. Em geral, o modelo hydrotime descreveu bem o comportamento germinativo, principalmente em temperaturas altas. Este trabalho mostra que o modelo pode ser usado para descrever a germinação de feijão comum em potenciais de água reduzidos, e como ferramenta para identificar genótipos de feijoeiro tolerantes à seca.

Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

Fusion of melanoma cells using a modified phytohaemagglutinin-ECM830 fusion method / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study melanoma cell fusion and find a highly efficient fusion method for tumor cells.</p><p><b>METHODS</b>Melanoma cells were labeled with green fluorescent protein and red fluorescent protein, respectively, and fused by a modified phytohaemagglutinin (PHA)-ECM830 fusion method. Melanoma fusion cells were selected by the fluorescence activated cell sorting. DNA content was determined by propidium iodide staining.</p><p><b>RESULTS</b>Melanoma cells were labeled with green fluorescent protein and red fluorescent protein markers and successfully fused through PHA-ECM830 fusion method. The fusion efficiency (7.18%) was much higher compared with ECM830 electricfusion method (0.50%). Melanoma fusion cells were successfully obtained by the fluorescence activated cell sorting.DNA content was doubled in melanoma fusion cells compared to B16-F10 melanoma cells. The proliferation rate of melanoma fusion cells was significantly decreased in vitro and in vivo.</p><p><b>CONCLUSIONS</b>We successfully obtained the melanoma fusion cells by the improved PHA-ECM830 fusion method. The proliferation rate of melanoma fusion cells dramatically decreases.</p>

Philadelphia chromosome detection in chronic myeloid leukemia: Utility of phytohemagglutinin-stimulated peripheral blood culture.

Background: The conventional cytogenetic approach to demonstrate Philadelphia (Ph) chromosome at times does not yield enough number of metaphases or are of suboptimal quality. Further, the rapid molecular tests have completely pushed this simple technique into disrepute. Aims: This study aimed to evaluate usefulness of phytohemagglutinin (PHA)-stimulated peripheral blood culture for detection of Ph chromosome in chronic myeloid leukemia (CML) patients. Materials and Methods: Fifty-six patients, including 11 newly diagnosed cases of CML and 45 patients of CML on imatinib therapy showing the presence of Ph chromosome in unstimulated samples, were included in the study. Cytogenetic analysis was done on unstimulated samples, i.e. bone marrow aspirate, 24- and 48-h peripheral blood culture, and compared with PHA-stimulated 72-h peripheral blood culture. Results: The preparations from PHA-stimulated peripheral blood culture samples in all 56 patients yielded high number of good-quality metaphases. All the 11 (100%) newly diagnosed patients and 39/45 (87%) of the patients on imatinib therapy showed the presence of Ph chromosome in PHA-stimulated samples. Addition of PHA-stimulated 72-h peripheral blood culture preparation can be of use for increasing the diagnostic yield in cases of CML with suboptimal results on conventional cytogenetics from bone marrow aspirate sample.

Immunosuppression of human amniotic mesenchymal cells on allogeneic peripheral blood lymphocytes / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the immunoregulatory effects of human amniotic mesenchymal cells (hAMCs) on allogeneic peripheral blood lymphocytes.</p><p><b>METHODS</b>The hAMCs were isolated from abandoned human amnion. Peripheral blood mononuclear lymphocytes (PBMLs) were separated from healthy donors by density gradient centrifugation. Then, PBMLs were treated with phytohemagglutinin (PHA) and different concentrations of hAMCs. Proliferation effect of PBMLs was tested using MTS assay, and production of IFN-gamma and TNF-alpha by PBMLs was detected by ELISA.</p><p><b>RESULTS</b>hAMCs could remarkably inhibit the lymphocytes proliferation. When the ratios of hAMCs to PBMLs were 0.05: 1, 0.10 :1, 0.20: 1, the inhibitory rates of PBMLs proliferation were 16.91%, 20.83% and 28.19%, respectively. HAMCs also decreased the production of IFN-gamma and TNF-alpha by PBMLs in a dose-dependent manner (P<0.01).</p><p><b>CONCLUSIONS</b>HAMCs could inhibit the proliferation of allogeneic lymphocytes and reduce secretion of IFN-gamma and TNF-alpha, which might be one of the mechanism for prevention and remission of transplant rejection.</p>

Screening for cytotoxic defects with flow cytometric detection of CD107α on natural killer cells and cytotoxic lymphocyte cells / 中华儿科杂志

<p><b>OBJECTIVE</b>To establish a novel flow cytometry-based assay for measuring the expression of lysosomal-associated membrane protein 1 (LAMP-1, CD107α) on the cell surface of natural killer (NK) cells and cytotoxic T lymphocyte (CTL) and evaluate the screening value of this assay for cytotoxic defects-related diseases such as familial hemophagocytic lymphopro-liferative (FHL) syndrome.</p><p><b>METHOD</b>Three suspected Chediak-Higashi Syndrome (CHS) patients, three suspected FHL patients and 10 healthy children were enrolled in the study from October 2010 to June 2011. Their PBMCs were separated and activated overnight with IL-2. After the granule release of NK cells activated by phytohemagglutinin (PHA) and CD8+T cells by anti-CD3, the CD107α expression were analyzed by flow cytometry. The peripheral blood DNA and RNA of the patients were extracted to analyze the pathogenic genes via DNA-PCR/RT-PCR and direct sequencing.</p><p><b>RESULT</b>The CD107α expression on CTL in the ten healthy children significantly increased after activation by anti-CD3 [(0.18 ± 0.07)% vs. (4.47 ± 2.36)%, P < 0.05] and NK cells after activation by PHA [(0.27 ± 0.07)% vs. (5.80 ± 2.83)%, P < 0.05]. The frequency of CD107α-expression NK cells in three suspected CHS after activation was significantly elevated when compared with the healthy control [0.5%, 0.6% vs. (5.80 ± 2.83)%] except patient 2. After the anti-CD3 activation, the frequency of CD107α expression on CTL cells also showed no significant difference [0.3%, 0.9%, 0.2% vs. (4.47 ± 2.36)%] in three patients. All of their mean fluorescence intensity (MFI) showed the same trend. Patient 1 and 3 were identified to have LYST mutations (Patient 1: c.5411-5414 del TTTC, L1741fsX1758 and c.7975 C > T, R2596X; Patient 3: c.4863G > A, R1563H and c.5392-5393delAA, E1739fsX1756). There was no mutation identified in the LYST gene for patient 2. CD107α expression of NK cells and CTL in the suspected FHL patients and in mirror of these findings, no underlying gene variation of PRF, MUNC13-4 and STX11 were identified.</p><p><b>CONCLUSION</b>We developed a method to quantitatively assess cytotoxicity of the NK cells and CTL by measuring the expression of CD107α on the cell membrane, which appeared to be an effective and rapid screening test for cytotoxic defects-related diseases such as FHL and other HLH secondary to primary immunodeficiency.</p>

Cord Blood Cellular Proliferative Response as a Predictive Factor for Atopic Dermatitis at 12 Months

Since the risk of developing allergic disease increases in individuals exposed to allergens previously, even during the neonatal period, the immunologic status of a fetus may be important in the subsequent development of allergy. We evaluated the fetal factors to predict atopic dermatitis (AD) at 12 months in 412 infants of a COhort for Childhood Origin of Asthma and Allergic Diseases (COCOA) in the general Korean population. Cord blood mononuclear cells (CBMCs) were stimulated with ovalbumin and phytohemagglutinin and cellular proliferative response and concentrations of interleukin-13 and interferon-gamma, were measured. The risk of developing AD was greater in boys than girls (OR 1.97, 95% CI 1.26-3.09), infants delivered by cesarean section than vaginally (OR 1.93, 95% CI 1.14-3.26) and infants with than without parental history of AD (OR 2.34, 95% CI 1.29-4.24). The CBMC proliferative response to phytohemagglutinin stimulation was higher in infants with than without AD (P = 0.048), but no difference was observed in ovalbumin-stimulated cells (P = 0.771). Risk factors for the development of AD at 12 months include male gender, delivery by cesarean section and parental history of AD. Increased CBMC proliferative response to phytohemagglutinin stimulation may predict the development of AD at 12 months.

Cord Blood Cellular Proliferative Response as a Predictive Factor for Atopic Dermatitis at 12 Months

Since the risk of developing allergic disease increases in individuals exposed to allergens previously, even during the neonatal period, the immunologic status of a fetus may be important in the subsequent development of allergy. We evaluated the fetal factors to predict atopic dermatitis (AD) at 12 months in 412 infants of a COhort for Childhood Origin of Asthma and Allergic Diseases (COCOA) in the general Korean population. Cord blood mononuclear cells (CBMCs) were stimulated with ovalbumin and phytohemagglutinin and cellular proliferative response and concentrations of interleukin-13 and interferon-gamma, were measured. The risk of developing AD was greater in boys than girls (OR 1.97, 95% CI 1.26-3.09), infants delivered by cesarean section than vaginally (OR 1.93, 95% CI 1.14-3.26) and infants with than without parental history of AD (OR 2.34, 95% CI 1.29-4.24). The CBMC proliferative response to phytohemagglutinin stimulation was higher in infants with than without AD (P = 0.048), but no difference was observed in ovalbumin-stimulated cells (P = 0.771). Risk factors for the development of AD at 12 months include male gender, delivery by cesarean section and parental history of AD. Increased CBMC proliferative response to phytohemagglutinin stimulation may predict the development of AD at 12 months.

In vitro soluble cd30 levels in patients with chronic stable coronary artery disease

The CD30 antigen seems to play a costimulatory role in maintaining the physiological balance between T-helper [Th] l/Th2 immune responses. In this study, plasma and in vitro soluble CD30 [sCD30] secretion was investigated in patients with coronary artery disease [CAD] as a plausible marker of dysregulated immune response. Twenty one patients with angiographically confirmed CAD and 31 healthy controls took part in this study. The levels of the activation marker sCD30 were determined in plasma and phytohaemagglutinin [PHA]-stimulated and unstimulated peripheral blood mononuclear cell cultures by ELISA. Plasma sCD30 levels did not differ significantly between the patients and controls. However, spontaneous sCD30 secretion was significantly lower in patients with CAD compared to controls [p < 0.001]. The soluble CD30 levels were significantly increased in the supernatant of PHA-stimulated PBMCs compared to unstimulated cultures in both groups of patients and controls [p < 0.001]. PHA-stimulated sCD30 secretion was found to be lower in patients compared to controls; however, the difference was not statistically significant. Plasma sCD30 levels were not statistically different in patients with chronic stable CAD, a well-known Thl-mediated disease, compared to controls; whereas decreased spontaneous and PHA-stimulated sCD30 secretion in patients with CAD might indicate the progressive shift towards a Thl immune response

Effect of phytohemagglutinin (PHA) from Yunnan white kidney bean on development of mouse embryos / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of different concentration of phytohemagglutinin (PHA) on mouse embryo development.</p><p><b>METHOD</b>In experiment 1, crude and purified PHA extracted from Yunnan white kidney bean with different concentration were added into M16 culture medium, the final concentration of PHA were: 50, 100, 200, 500, 1 000, 2 000 and 5 000 mg x L(-1) respectively. 2-cell stage embryos were collected and cultured in PHA containing or control medium for 72-96 h and their development were recorded. In experiment 2, different stage of embryos from 1-cell to blastocyst were treated by different concentrations of PHA same as experiment 1 and 10 000 mg x L(-1) in culture medium for 24 h before washing and cultured in M16 + PVA without PHA to blastocyst or hatching blastocyst stage.</p><p><b>RESULT</b>Low concentrations PHA at 50-100 mg x L(-1) promoted embryo development and increased the number of blastocyst stage embryos. In contrast, high concentrations of PHA (> 1 000 mg x L(-1)) blocked the embryos development from 1-cell to blastocyst stage and showed apoptosis morphology or death.</p><p><b>CONCLUSION</b>Depending on the concentrations, PHA from white kidney bean shown promotion or inhibition on mouse embryo development. 1-cell stage embryo shown more sensitive to PHA treatment than that of later stage embryos. Pretreatment 24 h in PHA containing medium can influence the further development of embryos. Low concentrations of PHA is benefit to embryo development, but high concentrations of PHA (> 1 000 mg x L(-1)) will block of the development of embryos.</p>

Fontes e doses de nitrogênio em cobertura no feijoeiro de inverno irrigado no sistema plantio direto / Sources and doses of sidedressing nitrogen on irrigated common bean cropped in winter in no tillage system

O feijoeiro é uma leguminosa de grande importância na economia brasileira, e o nitrogênio é o nutriente absorvido em maior quantidade. O manejo da adubação nitrogenada é de extrema importância no sentido de oferecer maior viabilidade econômica, além de aumentar a eficiência da planta na utilização dos recursos disponíveis. O objetivo do trabalho foi estudar o efeito de fontes e doses de nitrogênio em cobertura no desenvolvimento e produtividade do feijoeiro de inverno no sistema plantio direto, correlacionado com uma análise econômica simples. O delineamento experimental foi de blocos casualizados em esquema fatorial 3x3, constituídos pela combinação de três fontes de nitrogênio (sulfato de amônio, uréia e mistura - sulfato de amônio ½ do N + uréia ½ do N) e diferentes doses de nitrogênio em cobertura (zero, 40 e 80 kg ha-1, aplicado na fase V4-3), com quatro repetições. O projeto foi conduzido no município de Selvíria (MS), no período de outono-inverno de 2004. O solo do local é um LATOSSOLO VERMELHO Distrófico argiloso. Conclui-se que independente da fonte de N, o aumento da adubação nitrogenada proporciona incremento na produtividade do feijoeiro de inverno até a dose de 80 kg ha-1, sendo que esta proporciona em média, aumento de 25 % na produtividade comparado com a testemunha (sem N em cobertura). A uréia é a fonte de nitrogênio de maior eficiência econômica.

The common bean is a leguminous of great importance in the Brazilian economy and nitrogen is the taken up nutrient in larger amount. Nitrogen fertilization management is of extreme importance to offer larger economical viability, besides increasing the efficiency of plant in the use of the available resources. The objective this study was to evaluate the effect of sources and doses of sidedressing nitrogen in the development and yield of winter common bean in no tillage system, as well as evaluate its economical viability. A randomized blocks design was used, in a factorial scheme 3x3 with 9 treatments constituted by three sources of nitrogen (ammonium sulphate, urea and ammonium sulphate ½ of N + urea ½ of N, applied at V4-3 stadium) and different doses of sidedressing nitrogen (0, 40, 80 kg ha-1) in four replications. The study was conducted in Selvíria county, MS State in 2004 in no season crop period, in a dystrophic Haplustox soil. The conclusion: independent of nitrogen source, nitrogen fertilization increasing provides increment in yield of winter common bean up to dose 80 kg/ha, and this provides, on average, an increase of 20% in yield compared with control (without sidedressing nitrogen). The urea is the nitrogen source of larger economical efficiency