Two new prenylated 2-arylbenzofurans from roots of Artocarpus heterophyllus and their anti-respiratory burst activities / 中国中药杂志

Two prenylated 2-arylbenzofurans were isolated from roots of Artocarpus heterophyllus, with a combination of various chromatographic approaches, including ODS, MCI, Sephadex LH-20, and semipreparative high performance liquid chromatography(HPLC). They were identified as 5-[6-hydroxy-4-methoxy-5,7-bis(3-methylbut-2-enyl)benzofuran-2-yl]-1,3-benzenediol(1) and 5-[2H,9H-2,2,9,9-tetramethyl-furo[2,3-f]pyrano[2,3-h][1]benzopyran-6-yl]-1,3-benzenediol(2) with spectroscopic methods, such as HR-ESI-MS, IR, 1D NMR, and 2D NMR, and named artoheterins B(1) and C(2), respectively. The anti-respiratory burst activities of the two compounds were evaluated with rat polymorphonuclear neutrophils(PMNs) stimulated by phorbol 12-myristate 13-acetate(PMA). The results showed that 1 and 2 exhibited significant inhibitory effect on respiratory burst of PMNs with IC_(50) values of 0.27 and 1.53 μmol·L~(-1), respectively.

Enfermedad granulomatosa crónica: fisiopatología genética y molecular. Pruebas diagnósticas vigentes / Chronic granulomatous disease: genetic and molecular pathophysiology. Effective diagnostic tests

Introducción: La enfermedad granulomatosa crónica es una inmunodeficiencia primaria congénita del sistema inmune innato, originada por defectos en el complejo enzimático nicotinamida adenina dinucleótido fosfato oxidasa presente en células fagocíticas. Estos defectos funcionales causan incapacidad para producir especies reactivas del oxígeno en los fagocitos, que afectan la eliminación de algunos microorganismos patógenos dentro del fagolisosoma. El diagnóstico de esta enfermedad se realiza actualmente mediante la prueba de 1,2,3-dihidrorodamina asistida por citometría de flujo multiparamétrica, o la tinción de fagocitos con nitroazul de tetrazolio asistida por microscopio óptico. Objetivos: Describir los aspectos fisiopatológicos y moleculares de la enfermedad granulomatosa crónica; y discutir aspectos relacionados con las pruebas de diagnóstico antes mencionadas. Métodos: Se realizó una investigación bibliográfica-documental a partir de artículos científicos publicados desde 1933 hasta 2018, para ello fueron consultadas las bases de datos SciELO, PubMed y Springer. Desarrollo: Se exponen las características fisiopatológicas de la enfermedad granulomatosa crónica, así como la relación entre las mutaciones genéticas más abundantes en la población afectada y la gravedad de las manifestaciones clínicas que presentan los pacientes. Además, se analizan críticamente los beneficios y las deficiencias de dos técnicas que se utilizan actualmente para diagnosticar la enfermedad. Conclusiones: La enfermedad granulomatosa crónica puede generar consecuencias inmunológicas e inflamatorias graves, que se hallan en consonancia con las características genéticas expresadas en el complejo enzimático dañado. El diagnóstico de la enfermedad resulta más confiable, exhaustivo y específico, mediante la citometría de flujo y su prueba de 1,2,3-dihidrorodamina(AU)

Introduction: Chronic granulomatous disease is a congenital primary immunodeficiency of the innate immune system, caused by defects in the nicotinamide adenine dinucleotide phosphate oxidase enzyme complex present in phagocytic cells. These functional defects cause inability to produce reactive oxygen species in phagocytes, affecting the elimination of some pathogenic microorganisms within the phagolysosome. The diagnosis of this disease is currently made by means of the 1,2,3-dihydrorodamine test assisted by multiparametric flow cytometry, or the staining of phagocytes with nitro-blue tetrazolium assisted by light microscopy. Objectives: To characterize molecular and pathophysiologically the chronic granulomatous disease; and to discuss aspects related to the aforementioned diagnostic tests. Methods: In this work, a bibliographic-documentary research was carried out from scientific articles published from 1933 to 2018, for which the SciELO, PubMed and Springer databases were consulted. Development: The pathophysiological characteristics of chronic granulomatous disease are exposed, as well as the relationship between the most abundant genetic mutations in the affected population, and the severity of the clinical manifestations presented by the patients. In addition, the benefits and deficiencies of two techniques currently used to diagnose the disease are critically analyzed. Conclusions: Chronic granulomatous disease can generate severe immunological and inflammatory consequences, which are in line with the genetic characteristics expressed in the damaged enzyme complex. The diagnosis of the disease is more reliable, exhaustive and specific, using flow cytometry and its 1,2,3-dihydrorodamine test(AU)

Antepenultimate residue at the C-terminus of NADPH oxidase RBOHD is critical for its function in the production of reactive oxygen species in Arabidopsis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Production of reactive oxygen species (ROS) is a conserved immune response primarily mediated by NADPH oxidases (NOXs), also known in plants as respiratory burst oxidase homologs (RBOHs). Most microbe-associated molecular patterns (MAMPs) trigger a very fast and transient ROS burst in plants. However, recently, we found that lipopolysaccharides (LPS), a typical bacterial MAMP, triggered a biphasic ROS burst. In this study, we isolated mutants defective in LPS-triggered biphasic ROS burst (delt) in Arabidopsis, and cloned the DELT1 gene that was shown to encode RBOHD. In the delt1-2 allele, the antepenultimate residue, glutamic acid (E919), at the C-terminus of RBOHD was mutated to lysine (K). E919 is a highly conserved residue in NADPH oxidases, and a mutation of the corresponding residue E568 in human NOX2 has been reported to be one of the causes of chronic granulomatous disease. Consistently, we found that residue E919 was indispensable for RBOHD function in the MAMP-induced ROS burst and stomatal closure. It has been suggested that the mutation of this residue in other NADPH oxidases impairs the protein’s stability and complex assembly. However, we found that the E919K mutation did not affect RBOHD protein abundance or the ability of protein association, suggesting that the residue E919 in RBOHD might have a regulatory mechanism different from that of other NOXs. Taken together, our results confirm that the antepenultimate residue E is critical for NADPH oxidases and provide a new insight into the regulatory mechanisms of RBOHD.

Effect of hypobaric hypoxia on killing functions and secreting IL-6,TNF-α of peritoneal macrophages in mice / 中国免疫学杂志

Objective:To explore the influence of high altitude hypoxia on the phagocytosis and killing functions of peritoneal macrophages in mice by establishing mouse model in high altitude hypoxic environment.Methods:①After exposure of mice to an altitude of 4 200 m,2 200 m and 400 m for 30 d respectively,flow cytometry was used to detect the phagocytosis and killing functions of peritoneal macrophages on staphylococcus aureus labeled with FITC.②The respiratory burst level of the cultured macrophage in mice was detected in 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe method. ③The concentration of NO2- as a stable oxidative metabolite of NO in the supernatant of the cultured macrophages was measured with ELISA kits;④The release level of IL-6 and TNF-α in the cultured mice macrophage supernatant was also determined with ELISA kits.Results:After exposed under high altitude hypoxia for 30 d,compared with the control group (400 m),the phagocytosis,respiratory burst level and NO release in high altitude groups (4 200 m and 2 200 m) were all than those in the control group(400 m) (P<0.05).While the concentration of IL-6 and TNF-α in the Mφ cultured supernatant showed an obvious increase (P<0.05).Conclusion:Exposure under high altitude hypoxia (at altitude of 4 200 m and 2 200 m) for 30 d compromised the phagocytosis and oxygen dependent cytolyticactivity functions,and also raised the cytokines secretion level of IL-6 and TNF-α in Mφ,thereby affecting the innate immune response ability of Mφ in body.

Estallido respiratorio de neutrófilos por las técnicas del nitroazul tetrazolio (NBT) y dihidrorodamina (DHR) en niños con sospecha clínica de enfermedad granulomatosa crónica (EGC)

Introducción: El estallido respiratorio (ER) de neutrófilos es fundamental para la defensa contra infecciones, proceso ausente o ineficaz en la EGC, una inmunodeficiencia primaria (IDP) diagnosticada mediante la prueba del NBT. Actualmente se destacan las técnicas por citometría de flujo como la DHR, realizada únicamente en el Instituto de Investigaciones en Ciencias de la Salud (IICS), habiendo sido aplicada sólo en niños sanos. Objetivo: Evaluar el ER de neutrófilos por las técnicas NBT y DHR en niños con sospecha clínica de EGC y describir sus características clínico-demográficas. Materiales y Métodos: Se incluyeron 36 niños de ambos sexos, menores de 17 años de edad, remitidos al IICS entre el 2014-2015 por médicos especialistas. Se extrajo sangre para evaluación del ER de neutrófilos y se aplicó un cuestionario. Resultados: La mediana de edad fue de 4 años, 56% varones. Predominaron los pacientes hospitalizados, la sepsis y forunculosis cutánea fueron las manifestaciones clínicas más frecuentes y un 72% presentó recurrencia de infecciones con mediana de 3 episodios/año. El promedio para el IE de neutrófilos fue de 38,1±13,7 en el ensayo DHR, y 87±17% de activación para la prueba del NBT. En 8 pacientes los valores fueron inferiores al considerado normal y en un niño se confirmó EGC, observándose un patrón de herencia ligada al X. Conclusión: La evaluación del ER de neutrófilos permitió detectar un caso de EGC, determinándose el patrón hereditario mediante la técnica DHR por primera vez en el país. Es esencial el empleo de herramientas diagnósticas disponibles en niños con sospecha clínica de IDPs, para la detección y tratamiento oportunos que mejoran su calidad de vida y reducen la mortalidad.

Introduction: The neutrophil`s respiratory burst (RB) is essential for the defense against infections, this process is absent or ineffective in the CGD, a primary immunodeficiency (PID) diagnosed by the NBT test. Techniques that used flow cytometry such as DHR, performed only at the Instituto de Investigaciones en Ciencias de la Salud (IICS), currently stand out, having been applied only to healthy children. Objective: To evaluate the neutrophil`s RB using the NBT and DHR techniques in children with clinical suspicion of CGD and to describe their clinical and demographic characteristics. Materials and methods: 36 children of both sexes, with less than 17 years of age, that were referred to the IICS by specialists physicians between the years 2014-2015 were included. A blood sample was obtained to evaluate the neutrophil`s RB and a questionnaire was applied. Results: The median age was of 4 years and 56 % were males. Predominantly the patients were hospitalized, being sepsis and cutanueos furunculosis the most frequent clinical manifestations and a 72 % presented recurrent infection with a median of 3 episodes/year. The average for the neutrophil´s stimulation index (EI) was 38,1±13,7 with the DHR test, and 87±17% of activation for the NBT test. In 8 patients the values obtained were below the ones considered as normal and in one child CGD was confirmed, in which an X-linked inheritance pattern was observed. Conclusion: The evaluation of the neutrophil`s RB allowed the detection of one case of CGD, and the inheritance pattern was determined by the DHR test for the first time in our country. The use of available diagnostic tools in children with clinical suspicion of PID is essential for the appropriate detection and treatment that improve the quality of life and reduce mortality.

The effect of the timing of ERCP treatment option on the respiratory burst and levels of inflammatory cytokines of patients with acute obstructive suppurative cholangitis / 国际检验医学杂志

Objective To investigate the effect of the timing of endoscopic retrograde cholangiopancreatography(ERCP)treat-ment option on the respiratory burst and inflammatory factor level for acute obstructive suppurative cholangitis(AOSC)patients. Methods A total of 98 patients with AOSC who were treated in our hospital from January 2013 to June 2016 were retrospectively analyzed.The patients were divided into A group and B group according to the time of ERCP operation.42 cases in A group were received ERCP within 6 h after admission.56 cases in B group were received ERCP in 6-24 h after admission.,The serum levels of inflammatory cytokines were compared between the two groups before and after treatment.The apoptosis rate and respiratory burst rate of peripheral hematoma were compared between the two groups before and after treatment.The complications and death rate of the patients in the two groups were recorded.Results After treatment,the levels of TNF-α and IL-6 in the A group were signifi-cantly lower than those in the B group and the IL-10 level was significantly higher in the B group(P<0.05).After treatment,the rate of neutrophil apoptosis in the A group was significantly higher than that in the B group(P<0.05),and the respiratory burst level in the A group was significantly lower than that in the B group(P<0.05).The incidence number of complications and death in patients in group A was significantly lower than that in group B(P< 0.05).Conclusion ERCP treatment for patients with AOSC should be performed as early as possible and as soon as possible.Early ERCP treatment is safe and effective and can alleviate the inflammatory reaction of patients,improve the survival rate of patients,and is worthy of clinical promotion.

The neutrophil respiratory burst in newborns / 国际儿科学杂志

Neutrophil respiratory burst is like a double-edged sword.On one hand, newborns rely mainly on neutrophil against infection.On the other hand, newborns are succeptible to be damaged by reactive oxygen species.Studies have already suggested that reactive oxygen species have relationship with neonatal diseases, such as sepsis,bronchopulmonary dysplasia,but it still remains controversial.This paper provides an overview of the measurement of neutrophil respiratory burst, and the relationship of neutrophil respiratory burst with neonatal sepsis,as well as with bronchopulmonary dysplasia.

Role of activin A on regulation of mouse neutrophil function / 中国免疫学杂志

Objective:To study the role of activin A in regulation of neutrophil function by detecting activin receptor expression and cellular activities.Methods:Peritoneal neutrophils were isolated in mouse.After the neutrophils were stimulated with activin A,the expression of ActRⅡA on neutrophils was examined by immunofluorescence and flow cytometry.Expression of Smad3 in neutrophils was analyzed by Western blot.Assays of neutrophils function were performed by detecting respiratory burst, production of NO and phagocytosis.Results:The isolated cells were composed of more than 90% peritoneal neutrophils.ActRⅡA was expressed on mouse neutrophils and Gr-1/ActRⅡA double-positive cells were 41.1%.Activin A promoted Smad3 phosphorylation in neutrophils,increased the production of ROS and O2-(P<0.05),enhanced secretion of NO and phagocytosis of mouse neutrophils(P<0.01),and promoted fluorescent microsphere phagocytosis of neutrophils by flow cytometry ( P<0.01 ) .Conclusion: Activin receptor and activin signaling protein were expressed on mouse neutrophils,activin A might play an important regulatory role in activation and function of neutrophils.

Effect of dexmedetomidine on apoptosis and respiratory burst of polymorphonuclear neutrophils in vitro / 实用医学杂志

Objective To investigate the effect of dexmedetomidine (Dex) on the apoptosis and respiratory burst of human polymorphonuclear neutrophils (PMN) in vitro. Methods The peripheral venous blood was collected from healthy volunteers. Isolation of PMN was performed by using the discontinuous plasma-Percoll gradient technique. PMN was randomLy divided into four groups: control group and three different concentrations of Dex (1 ng/mL,10 ng/mL,100 ng/mL) groups. PMN was cultured in enriched RPMI-1640 media at 2 × 106/mL for 24 h. The caspase-3 activity, apoptosis rate and respiratory burst of PMN were detected at 2 and 24 h. Results Compared with the control group, the PMN which was treated with three different concentrations of Dex showed no significant difference in caspase-3 activity, apoptosis rate and respiratory burst of PMN after 2 h of incubation. Compared with the control group, 1 ng/mL concentration of Dex did not affect the caspase-3 activity, apoptosis and respiratory burst of PMN cultured for 24 h. However, 10 ng/mL and 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incubation. Compared with 10 ng/mL Dex group, 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incutation. Conclusion Clinically relevant concentration of Dex does not affect apoptosis and respiratory burst of PMN, while high concentration of dexmedetomidine can induce apoptosis of PMN.

Disease resistance of pacu Piaractus mesopotamicus (Holmberg, 1887) fed with β-glucan / Resistência à doença em pacu, Piaractus mesopotamicus (Holmberg, 1887), alimentados com β-glucana

Effects of β-glucan on innate immune responses and survival were studied in pacu experimentally infected with Aeromonas hydrophila. Fish fed diets containing 0, 0.1% and 1% β-glucan were injected with A. hydrophila. β-glucan enhanced fish survival in both treated groups (26.7% and 21.2% of the control, respectively). Leukocyte respiratory burst and alternative complement pathway activities were elevated after bacterial challenge regardless the β-glucan concentration. Lysozyme activity was higher after infection and showed a gradual increase as β-glucan concentration increased. A significant elevation in WBC count was observed either after bacterial challenge or by influence of β-glucan separately. The same response was observed in the number of thrombocytes, lymphocytes, eosinophils, LG-PAS positive cell and monocytes. It can be concluded that feeding pacu with β-glucan can increase protection against A. hydrophila, due to changes in non-specific immune responses.

Os efeitos da β-glucana sobre as respostas imunes inatas e a sobrevivência foram estudados em pacu experimentalmente infectado com Aeromonas hydrophila. Peixes alimentados com dietas contendo 0,1% e 1% de β-glucana foram injetados com 1 × 108 CFU de A. hydrophila após 7 dias de alimentação. A sobrevivência de peixes foi maior nos dois grupos tratados em comparação ao grupo controle (26,7% e 21,2%, respectivamente). A atividade respiratória de leucócitos e a atividade hemolítica do complemento – via alternativa estavam elevadas após desafio bacteriano independentemente da concentração de β-glucana. A atividade de lisozima foi maior após a infecção e mostrou um aumento gradual de acordo com a concentração do imunoestimulante. Observou-se um aumento significativo na contagem de leucócitos totais após o desafio bacteriano e influência de β-glucana. A mesma resposta foi observada para trombócitos, linfócitos, eosinófilos, leucócito PAS positivo e monócitos. Com exceção de neutrófilos, que diminuíram frente ao mais alto nível do imunoestimulante e não se alteraram após a infecção, as outras células aumentaram após a exposição à A. hydrophila. A β-glucana não afetou os níveis de proteína total do soro, que aumentaram após o desafio bacteriano. Conclui-se que a administração de β-glucana em pacu pode aumentar a proteção contra A. hydrophila, por alterações nas respostas imunes de não-específicas.

Respiratory Burst Enzymes and Oxidant-antioxidant Status in Nigerian Children with Sickle Cell Disease.

Aim: To measure respiratory burst enzymes, pro-oxidants, antioxidants and red cell indices in Nigerian children with sickle cell disease (HbSS) below five years of age and compared with apparently healthy children with normal haemoglobin (HbAA). Method: A total of 45 subjects were recruited which included 23 children (age range 10 – 48 months) with HbSS and 22 children (age- and sex- matched) with HbAA. Blood samples were collected and red cell indices were determined using automated haematology analyser while serum superoxide dismutase (SOD), glutathione peroxidise (GSH-Px) and myeloperoxidase (MPO) activities were measured using ELISA kits. Serum malondialdehyde (MDA), hydrogen peroxide (H2O2), glutathione S transferase (GST), catalase (Cat), xanthine oxidase (XO) and glutathione (GSH) were measured with colorimetric techniques. MPO, SOD and Cat represented respiratory burst enzymes; MDA, H2O2 and XO were measured as pro-oxidants while GSH, GST and GSH-Px were the measured antioxidants. Results: Mean concentration of malondialdehyde was significantly reduced (5.56±1.12nmol/L vs. 6.46±1.11nmol/L, P=.04) in HbSS children compared with HbAA children. Similarly, mean serum activity of myeloperoxidase in HbSS children was significantly reduced compared with HbAA children (66.12±13.34U/mL vs 77.02±13.54U/mL, P=.03). However, there were no significant differences in mean concentration of serum glutathione, hydrogen peroxide; serum activities of glutathione peroxidase, catalase, superoxide dismutase, xanthine oxidase and glutathione S transferase in HbSS children compared with HbAA children Conclusion: HbSS children in this population did not demonstrate raised oxidative stress.

Increased oxygen consumption observed in phorbol 12-myristate 13-acetate stimulated human cultured promonocytic U937 cell lines treated with calcitriol and retinoic acid

OBJECTIVE@#To investigate the effect of phorbol 12-myristate 13-acetate (PMA) and formyl-methionyl-leucyl-phenylalanine (FMLP) on oxygen consumption of differentiated and non-differentiated immune cell lines by retinoic acid and calcitriol treatment which might be useful in subsequent elicitation of immunological action during immunosuppressive states.@*METHODS@#PMA and FMLP were used to artificially stimulate reactive oxygen production in cultured promonocytic U937 cell line. Paralleled samples of the cultured cells were separately prepared with calcitriol (1, 25- dihydroxyvitamin D3) and retinoic acid followed by a 72-hour incubation period. The rate of respiratory burst was measured using the Clark oxygen electrode.@*RESULTS@#The average increase in cell concentrations per mL observed was significantly higher in retinoic acid-treated cells (9×10(6) cells/mL) when compared with calcitriol-treated samples (4×10(6) cells/mL). There was a marked increase in oxygen consumption of the calcitriol-treated cell lines against the retinoic acid-treated ones. Exposure of differentiated U937 cells to PMA and FMLP increased significantly (P<0.05) in their oxygen consumption when compared with the control. PMA calcitriol-treated cells resulted in 55% oxygen consumption more than the control while FMLP oxygen consumption increased 78% by comparison with the control.@*CONCLUSIONS@#The result demonstrated that calcitriol may serve as a physiological promoter of normal differentiation of precursor cells which may exert an immunological action. This effect could elicit a marker potential and increase immune cell activity of the host especially in immunosuppressed diseased states.

Effects of glucose concentration on G6PD activity and respiratory burst of normal human′s neutrophils in vitro / 重庆医学

Objective To determine the effects of glucose concentration on G 6PD activity and respiratory burst of normal hu-man′s neutrophils in vitro .Methods Normal human′s neutrophils were cultured in different glucose concentration for 8 hours ,as-sayed G6PD activity by spectrophotometric method and determining ROS content by fluorescent probe DCFH-DA .Results G6PD activity and ROS of 15 mmol/L group and 25 mmol/L group were significant lower than before ,when the 5 mmol/L group and L-GLU group didn′t have significant change with time goes by .And G6PD activity and ROS of 25 mmol/L group were the lowest in all groups(P<0 .01) .Conclusion High glucose may induce G6PD activity decreased and cause respiratory burst dysfunction as a stimulating factor .The stimulation intensity was increased with the increase of glucose concentration .It′s the probable mechanism on susceptibility to infections in patients with diabetes mediated by dysfunction of respiratory burst in leucocyte .

Respiratory burst function of neutrophils in very low birth weight infants / 中华围产医学杂志

Objective To investigate the respiratory burst function of neutrophils in very low birth weight infants (VLBWI).Methods Twenty two VLBWI was divided into two groups:neonatal respiratory distress syndrome (NRDS) and non NRDS (11 in each).The respiratory burst function of neutrophils in the peripheral blood of VLBWI within 48 hours after birth was determined using the flow cytometrydihydrorhodamine 1,2,3 method before and after the chemical stimulation of phorbol-12-myrismte 14 acetate (PMA),and the gp91Phox was also measured in resting neutrophils by flow cytometry.Twenty healthy term neonates served as controls.Mann-Whitney U test was used for statistical analysis.Results Before the stimulation of PMA,the percentage of activated neutrophils of VLBWI [(49.10±20.19) ％] producing a respiratory burst was higher than that of term neonates [(18.73 ±6.81) ％] (Z--4.911,P=0.000),however,after the stimulation of PMA,the percentage of activated neutrophils of VLBWI [(96.58 ± 3.44) ％] was lower than that of term neonates [(99.20±0.62) ％] (Z--3.186,P=0.001),and the stimulation index (SI) of VLBWI (171.40 ± 103.35) was lower than that of term neonates (306.30 ± 138.47),with significant difference (Z=-3.413,P=0.001).The geometric mean of gp91Phox in VLBWI (21.66± 19.87) was higher compared with term neonates (19.60±8.03),however,the difference was not significant (P=0.350).The percentage of neutrophils that expressed gp91Phox [(56.11 ± 29.40) ％] was lower in VLBWI than that in term neonates [(80.14± 14.87) ％],with significant difference (Z=-2.374,P=0.018).Before the stimulation of PMA,the percentage of activated neutrophils of VLBWI with NRDS (63.40± 16.45) ％] was higher than that of VLBWI without NRDS [(34.80± 11.65) ％],with significant difference (Z=-3.382,P=0.001),the SI of VLBWI with NRDS (129.46 ± 75.36) was significantly lower than that of VLBWI without NRDS (213.35 ± 113.49) (Z=-2.331,P=0.020).Conclusions Neutrophils producing a respiratory burst in both VLBWI and term neonates are active without stimulation of PMA,while the phenomenon is more obvious in VLBWI.Neutrophils in VLBWI and term infants can be activated by the stimulation of PMA,and express gp91Phox.The activation and gp91Phox expression of neutrophils in VLBWI with NRDS tend to be lower than those in VLBWI without NRDS.

Report of a case of severe congenital neutropenia with literature review / 临床儿科杂志

Objective To investigate the clinical features and pathogenesis of severe congenital neutropenia (SCN) by detecting the gene mutation of a SCN patient suspected by clinical diagnosis. Methods The intravenous anticoagulant and clin-ical data and laboratory results of this child were collected;the phagocyte and oxidation function of neutrophils were evaluated by flow cytometry;ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4 genes were screened by PCR amplification and sequencing. Results The neutrophil function of this patient was normal; sequencing results revealed no mutation occurred in ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4;and granulocyte colony-stimulating factor (G-CSF) can obviously enhance the level of neutrophils. Conclusion SCN is a kind of genetic heterogeneity syndrome associated with multiple gene mutations, gene diag-nosis will contribute to understanding of the pathogenesis of the disease and provide theoretical basis for treatment. Though more and more pathogenic genes were found to be connected with SCN, the cases of unknown mutation still account for a large proportion of this disease.

Increased oxygen consumption observed in phorbol 12-myristate 13-acetate stimulated human cultured promonocytic U937 cell lines treated with calcitriol and retinoic acid

Objective: To investigate the effect of phorbol 12-myristate 13-acetate (PMA) and formyl-methionyl-leucyl-phenylalanine (FMLP) on oxygen consumption of differentiated and non-differentiated immune cell lines by retinoic acid and calcitriol treatment which might be useful in subsequent elicitation of immunological action during immunosuppressive states. Methods: PMA and FMLP were used to artificially stimulate reactive oxygen production in cultured promonocytic U937 cell line. Paralleled samples of the cultured cells were separately prepared with calcitriol (1, 25- dihydroxyvitamin D

Effects of intraoperative thermostasis on respiratory burst of polymorphonuclear neutrophil in patients undergoing radical operation for lung cancer / 中华麻醉学杂志

Objective To investigate the influence of intraoperative thermostasis over respiratory burst of polymorphonuclear neutrophils (PMNs) in patients undergoing radical operation for lung cancer.Methods Thirty-two ASA Ⅱ or Ⅲ patients scheduled for radical operation for lung cancer under general anesthesia were randomized into two groups ( n = 16 each): control group (Group C) and warming group (Group W). The patients in Group C were kept warm by routine measures such as using woollen blankets, while the patients in Group W were kept warm by force-air warming system and fluid warming device as soon as the patients were admitted to the operation room. Rectal and axillary temperatures were continuously monitored as the core and surface temperature, respectively. The core temperature was maintained at the preoperative level (baseline). Anesthesia was induced with midazolam, fentanyl and propofol. Tracheal intubation was facilitated with rocuronium. Anesthesia was maintained with isoflurane and nitrous oxide and intermittent i.v. boluses of fentanyl, midazolam and vecuronium. Venous blood samples were obtained before, during and at the end of surgery for normal blood analysis and respiratory burst of PMNs which included activated PMNs count and reactive oxygen species (ROS) production.Results (1) WBC and PMN counts were significantly increased during and after operation as compared with the baseline values before operation in both groups and there was no significant difference in WBC and PMN counts between the two groups. (2)Phorbol-12-myristate-13-acetate (PMA) stimulation resulted in higher intraoperative and postoperative activated PMN counts in both groups and higher postoperative ROS production in Group W. Postoperative ROS production was significantly higher in Group W than in Group C. (3) The PMN counts without stimulation activation during operation and intra- and post-operative ROS production were significantly decreased as compared with the baseline values before operation in Group C, while in Group W there was no significant difference in pre-, intra- and post-operative activated PMN counts and ROS production. The intraoperative PMN counts and intra- and post-operative ROS productions were significantly higher in Group W than in Group C.Conclusion Intraoperative thermostasis can effectively maintain activated PMN count and ROS production without stimulation and enhance ROS production with stimulation in patients undergoing radical operation for lung cancer.

Mediadores solubles liberados por la infección con Trypanosoma cruzi podrían desencadenar mecanismos de fisiopatogenia en la enfermedad de Chagas experimental / Soluble mediators in Trypanosoma cruzi infection could trigger mechanisms of pathogenesis in Chagas disease

El Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, afecta a cerca de 2.500.000 personas en nuestro país y 18 millones en América latina. Este parásito presenta en el hospedador dos estadios de importancia médica: la forma amastigote, intracelular, que se replica activamente, y la forma extracelular, el tripomastigote, que es infectante. Por ello, para controlar la infección se requiere una potente respuesta inmune humoral y celular, y es importante el resultado de la interacción hospedador­parásito en etapas tempranas de la infección. El 30% de los infectados desarrolla algún grado de patología, cardíaca o digestiva, a lo largo de los años. Algunos autores lo atribuyen a la acción directa del parásito, otros, a reacciones autoinmunes inducidas por el T. cruzi. El objetivo del presente trabajo fue estudiar en líquido peritoneal (sitio de infección), en un modelo experimental murino, algunos mediadores de respuesta innata en las primeras horas post infección con T. cruzi y la respuesta adaptativa al final de la fase aguda. Los resultados revelaron aumento del estallido respiratorio y síntesis de IL-6 desde las primeras horas. La concentración de óxido nítrico aumentó con la evolución de la infección mientras que la activación de arginasa se mantuvo controlada y fue importante la producción de inmunoglobulinas específicas. Los mecanismos estudiados estarían involucrados no sólo en la eliminación del T. cruzi, sino también en la fisiopatogenia de la infección. Estos hallazgos estimulan la continuación de estudios orientados a avanzar en el conocimiento de eventos inmunológicos tempranos para el desarrollo de nuevas terapéuticas profilácticas en el hombre.(AU)

Trypanosoma cruzi, ethiologic agent of Chagas disease, affects about 2.500.000 people in our country and 18 millon in Latin America. The parasite presents two stages of medical importance in the host, the amastigote, intracellular replicating form and the extracellular trypomastigote, the infective form. That is why the control of infection requires a strong humoral and cellular immune response, hence, it is very important the outcome of host-parasite interaction in the early stages of infection. 30% of infected people, develop some degree of pathology, cardiac or digestive in the chronic period of infection, attribute this to the direct action of the parasite, or to autoimmune reactions induced by Trypanosoma cruzi. The aim of this work was to study at local level, in a murine experimental model some mediators in the early infections with T. cruzi and the adaptive response at the end of the acute phase. The results revealed increased respiratory burst and synthesis of IL-6 since the first hours. Nitric oxide concentration increased with the progression of the infection, while the activation of arginase remained regulated and it was important the specific immunoglobulin production. These mediators would be involved in mechanisms of resistence to T. cruzi, but also in the pathogenia of the infection. These findings stimulate to go further in the knowledge of immunological early events directed to the development to therapeutic approaches in Chagas disease(AU)

Leukocytes respiratory burst and lysozyme level in pacu (Piaractus mesopotamicus Holmberg, 1887) / Atividade respiratória de leucócitos e nível de lisozima em pacu (Piaractus mesopotamicus)

Innate immune responses are useful to determine the health status of fish and to evaluate the effect of immunomodulatory substances in fish farming. Leukocytes respiratory burst was measured in pacu (Piaractus mesopotamicus) using chemiluminescence assay and nitroblue tetrazolium (NBT) reduction assay. The nitroblue tetrazolium reduction seemed more adequate than chemiluminescence assay for leukocytes oxidative burst determination, since it was difficult to isolate the blood leucocytes for chemiluminescence assay. Plasma and serum lysozyme were measured using a turbidimetric assay. The heating of serum and plasma samples (56 ºC for 30 minutes) for complement system inactivation darkened the plasma samples and interfered in the results. The lysozyme activity in serum was higher than in plasma, suggesting that serum samples are more appropriate for the analysis. This study established protocols that can be useful tools in the study of immune mechanisms of the tropical fish pacu.

Respostas imunológicas inatas são úteis para determinar o estado de saúde de peixes e avaliar o efeito de substâncias imunomoduladoras no cultivo destes animais. A atividade respiratória de leucócitos foi medida em pacu (Piaractus mesopotamicus) através de ensaio de quimioluminescência e ensaio de redução do nitroblue tetrazolium (NBT). O ensaio de redução do nitroblue tetrazolium pareceu mais adequado que o ensaio de quimioluminescência para determinação da atividade respiratória de leucócitos, uma vez que foi difícil isolar com êxito os leucócitos do sangue para o ensaio de quimioluminescência. Lisozima sérica e plasmática foram medidas por meio de ensaio turbidimétrico. Com o objetivo de inativar as proteínas do sistema complemento, as amostras de soro e plasma foram aquecidas (56 ºC por 30 minutos). Porém, este procedimento provocou a turvação das amostras de plasma e interferiu nos resultados. A atividade de lisozima no soro foi maior que no plasma, sugerindo que amostras de soro são mais apropriadas para esta análise. Este estudo estabeleceu protocolos que podem ser utilizados como ferramentas no estudo de mecanismos imunológicos do peixe tropical pacu.

Apoptosis, shape change and reactive oxygen species generation in human neutrophils exposed to olanzapine, an a typical antipsychotic drug

The effects of olanzapine (OLZ) on the viability and functioning of human polymorphonuclearcells (PMNs) are clearly opposite to those previously reported forclozapine (CLZ). In fact, after 4- or 24-h-treatment with 20-50 μM OLZ, a significant inhibition of the respiratory burst in PMNs activated with opsonized zimosanor phorbol myristate acetate (PMA) was observed, whereas the burst provoked byformyl-methionyl-leucyl-phenylalanine (fMLP) was only inhibited at 50 μM OLZ.Under the same conditions, spontaneous apoptosis was accelerated at 20-50 μMOLZ, while the exogenous addition of H2O2 resulted in the PMN apoptosis beingdose-dependently inhibited by OLZ in the entire range of concentrations. However,when H2O2 was intracellularly generated by treatment with PMA, the induced apoptosis was only decreased at 2 μM OLZ. Absorbance scans revealed that OLZis able to react with equimolar quantities of either H2O2 or HOCl. These results suggest that OLZ inhibits both ROS-induced PMN apoptosis and respiratory burst due to extracellular scavenging of released ROS.

Los efectos de olanzapine (olz) sobre la viabilidad y el funcionamiento de células humanas polimorfonucleares (pmn, por sus siglas en inglés) claramente son opuestosa los señalados para la clozapine (clz). En efecto, después de 4-24 h de tratamiento con 20-50 μM olz, se observó una inhibición significativa del estallido respiratorio en pmn activados con zimosan o con forbol acetato miristato, mientras que la inhibición provocada por el formil-metionil-leucil-fenilalanina fue sólo inhibida a 50μM de olz. En las mismas condiciones, la apoptosis espontánea se aceleró con 20-50μM olz, mientras que la adición exógena de H2O2 dio lugar a la apoptosis de pmn en dosis dependiente inhibida por olz en el rango entero de concentraciones. Sin embargo, cuando se generó H2O2 intracelular por tratamiento con pma, la apoptosis inducida se disminuyó solamente con 2 μM olz. Las exploraciones de los espectros de absorbancia revelaron que olz puede reaccionar con cantidades equimolares de H2O2 o de HOCL. Estos resultados sugieren que olz inhibe ambos tipos de apoptosis de pmn (la inducida por especies reactivas oxigenadas y por estallido respiratorio debido a atrapadores extracelulares de estas especies reactivas oxigenadas).