RESUMO
INTRODUCTION: Insurance coverage for oral valganciclovir (VGCV) began in Japan in April 2023 on the basis of results, including our clinical trials for symptomatic congenital cytomegalovirus (CMV) disease. The VGCV treatment is available throughout Japan, so clinicians must consider the likelihood of hearing improvement and the possibility of neutropenia before dosing. MATERIALS AND METHODS: We performed a substudy of an investigator-initiated, single-arm, prospective, multicenter, clinical trial in which 24 infants with symptomatic congenital CMV disease were orally administered 16 mg/kg VGCV twice daily for 6 months as an intervention. We examined the infants' baseline characteristics associated with improved hearing impairment or a severely reduced neutrophil count. RESULTS: Of the 24 patients, 4 had normal hearing on assessment of their ear with the best hearing. Hearing impairment improved in 14 patients and did not respond to VGCV treatment in 6 patients at the 6-month hearing assessment. CMV DNA levels in plasma at baseline were higher in patients in whom hearing did not respond to treatment. A neutrophil count <500/mm3 occurred in 5 (21%) patients for the first 6 weeks and in 8 (33%) patients for the first 6 months. A neutrophil count at screening and the lowest neutrophil count over the 6 months showed the highest correlation (r = 0.477, p = 0.019). CONCLUSIONS: Infants with a low plasma viral load at screening tend to have an improvement in hearing impairment. Clinicians should be aware of neutropenia during VGCV treatment particularly in patients with a low neutrophil count during screening.
Assuntos
Antivirais , Infecções por Citomegalovirus , Citomegalovirus , Neutropenia , Valganciclovir , Humanos , Valganciclovir/uso terapêutico , Valganciclovir/administração & dosagem , Infecções por Citomegalovirus/tratamento farmacológico , Infecções por Citomegalovirus/virologia , Antivirais/uso terapêutico , Antivirais/administração & dosagem , Antivirais/efeitos adversos , Feminino , Lactente , Masculino , Estudos Prospectivos , Administração Oral , Citomegalovirus/isolamento & purificação , Citomegalovirus/efeitos dos fármacos , Citomegalovirus/genética , Recém-Nascido , Japão , Resultado do Tratamento , Perda Auditiva/virologia , DNA Viral/sangue , Ganciclovir/análogos & derivados , Ganciclovir/administração & dosagem , Ganciclovir/uso terapêutico , Ganciclovir/efeitos adversos , Neutrófilos/efeitos dos fármacosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known as Nagphana, belongs to the Cactaceae family. It is traditionally used to treat various ailments including inflammation, gastric ulcers, diabetes, hepatitis, asthma, whooping cough and intestinal spasm. AIM OF THE STUDY: Despite its traditional use in various countries, detailed toxicological studies of O. dillenii cladode are few. Thus in the current study, toxicity of O. dillenii cladode derived methanol extract, fractions and its α-pyrones: opuntiol and opuntioside have been addressed. METHODS: The test agents were assessed using both in vitro and in vivo toxicity assays. MTT on human embryonic kidney cell line (HEK-293), tryphan blue exclusion in rat neutrophils, Cytokinesis-B block micronucleus (CBMN) in human lymphocytes and genomic DNA fragmentation using agarose gel electrophoresis were performed. In acute toxicity test, mice orally received extract (5 g/kg) for 7 days followed by measurements of relative organ weight, biochemical (blood profile, liver and kidney function test) and histological studies (liver and kidney) were carried out. Rat bone marrow micronucleus genotoxicity assay was also conducted. RESULTS: O. dillenii derived test agents were non-cytotoxic and had no effect on the integrity of DNA. Methanol extract (5 g/kg) orally administered in mice did not cause any significant change in relative organ weights, biochemical parameters and liver and kidney histology as compared to vehicle control. In parallel, extract did not stimulate micronuclei formation in rat bone marrow polychromatic erythrocytes. CONCLUSION: These results led to conclude that edible O. dillenii extract is non-toxic via the oral route and appears to be non-cyto-, hepato-, nephro- or genotoxic, thereby supporting its safe traditional use against various ailments. Therefore, opuntiol and opuntioside may serve as lead compounds in designing new drug(s) derived from edible plants.
Assuntos
Ácidos Cumáricos/toxicidade , Monossacarídeos/toxicidade , Opuntia/química , Extratos Vegetais/toxicidade , Animais , Ácidos Cumáricos/isolamento & purificação , Fragmentação do DNA/efeitos dos fármacos , Feminino , Células HEK293 , Humanos , Masculino , Metanol/química , Camundongos , Testes para Micronúcleos , Monossacarídeos/isolamento & purificação , Neutrófilos/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Pironas/isolamento & purificação , Pironas/toxicidade , Ratos , Ratos Sprague-Dawley , Testes de Toxicidade AgudaRESUMO
Decreased neutrophil function following administration of chemotherapy has been reported in dogs with lymphoma. The first objective of our study was to determine if neutrophil oxidative burst and phagocytic activity are affected by chemotherapy 7 to 10 days following initiation of treatment in dogs with lymphoma and non-lymphoma malignancies. The second objective was to determine if there is a correlation between neutrophil numbers and neutrophil function before or after initiation of chemotherapy. Flow cytometric assessment of neutrophil oxidative burst and phagocytosis following stimulation with Escherichia coli was performed in 9 dogs diagnosed with lymphoma and 17 non-lymphoma tumor-bearing dogs pre- and post-chemotherapy, as well as 14 tumor-free control dogs. Spearman rank correlation was performed to determine if blood neutrophil numbers and neutrophil function were significantly correlated. Lymphoma patients showed significantly reduced percentage neutrophil oxidative burst post-chemotherapy compared to healthy controls as well as compared to pre-chemotherapy values (P = 0.0022 and P = 0.0020, respectively). Lymphoma patients also exhibited significantly reduced neutrophil phagocytosis activity post-chemotherapy compared to controls and pre-chemotherapy values (P = 0.0016 and P = 0.014, respectively). Dogs with non-lymphoma malignancies also showed a significant decrease in both percentage oxidative burst and phagocytosis post-chemotherapy compared to pre-chemotherapy values (P = 0.00040 and P = 0.029, respectively). Neutrophil numbers and function were not significantly correlated. The results of the study suggest that chemotherapeutic treatment decreases neutrophil oxidative burst and phagocytic activity 7 to 10 days post-treatment in dogs with various malignancies. Furthermore, neutrophil numbers cannot be used to predict neutrophil function.
Une diminution de la fonction des neutrophiles après l'administration d'une chimiothérapie a été rapportée chez des chiens atteints de lymphome. Le premier objectif de notre étude était de déterminer si la stimulation oxydative des neutrophiles et l'activité phagocytaire sont affectées par la chimiothérapie 7 à 10 jours après le début du traitement chez les chiens atteints de lymphomes et de tumeurs malignes non lymphomateuses. Le deuxième objectif était de déterminer s'il existe une corrélation entre les nombres de neutrophiles et la fonction des neutrophiles avant ou après le début de la chimiothérapie. L'évaluation par cytométrie en flux de la stimulation oxydative des neutrophiles et de la phagocytose après stimulation par Escherichia coli a été réalisée chez neuf chiens diagnostiqués avec un lymphome et 17 chiens avec des tumeurs non lymphomateuses avant et après la chimiothérapie, ainsi que 14 chiens témoins sans tumeur. Une corrélation des rangs de Spearman a été effectuée pour déterminer si les nombres de neutrophiles sanguins et la fonction des neutrophiles étaient significativement corrélés. Les patients atteints de lymphome ont montré un pourcentage significativement réduit de stimulation oxydative des neutrophiles après la chimiothérapie par rapport aux témoins sains ainsi que par rapport aux valeurs pré-chimiothérapie (P = 0,0022 et P = 0,0020, respectivement). Les patients atteints de lymphome ont également présenté une activité de phagocytose par les neutrophiles significativement réduite après la chimiothérapie par rapport aux témoins et aux valeurs pré-chimiothérapie (P = 0,0016 et P = 0,014, respectivement). Les chiens atteints de tumeurs malignes non lymphomateuses ont également montré une diminution significative du pourcentage de stimulation oxydative et de la phagocytose post-chimiothérapie par rapport aux valeurs pré-chimiothérapie (P = 0,00040 et P = 0,029, respectivement). Les nombres et la fonction des neutrophiles n'étaient pas significativement corrélés. Les résultats de l'étude suggèrent que le traitement chimiothérapeutique diminue la poussée oxydative des neutrophiles et l'activité phagocytaire 7 à 10 jours après le traitement chez les chiens atteints de diverses tumeurs malignes. De plus, les nombres de neutrophiles ne peuvent pas être utilisés pour prédire la fonction des neutrophiles.(Traduit par Docteur Serge Messier).
Assuntos
Antineoplásicos/uso terapêutico , Doenças do Cão/tratamento farmacológico , Neoplasias/veterinária , Neutrófilos/efeitos dos fármacos , Animais , Antineoplásicos/efeitos adversos , Estudos de Casos e Controles , Cães , Escherichia coli , Feminino , Masculino , Fagocitose/fisiologia , Explosão Respiratória/efeitos dos fármacosRESUMO
This study was conducted to establish the toxicological profile of combination treatment with therapeutic HPV DNA vaccines (GX-188E) and the long-acting form of recombinant human interleukin-7 fused with hybrid Fc (IL-7hyFc). GX-188E was administered intramuscularly by electroporation with or without IL-7hyFc intravaginally once per 2 weeks for 8 weeks (five times) in female Sprague-Dawley rats. Because up-regulation of immune responses and migration of antigen-specific T cells in cervicoviginal tissue were predicted as therapeutic effects, we distinguished adverse effects from therapeutic effects based on the severity of the systemic immune response, reversibility of lymphoid tissue changes, target tissue damage, and off-target immune responses. We observed that the number of neutrophils was increased, and the number of lymphocytes was decreased in the blood. Further, myofiber degeneration, necrosis, fibroplasia, and cell infiltration were observed at the GX-188E administration site. These changes were fully or partially recovered over a 4-week period. Analysis of lymphocytes in spleen revealed that CD4+ T cells and total T cells decreased in rats treated with GX-188E in combination with a high dose of IL-7hyFc (1.25 mg/animal). However, these changes were not considered adverse because they were transient and may have been related to electroporation-mediated DNA delivery or the local migration of lymphocytes induced by IL-7. Therefore, the potential toxicity of the combination of GX-188E and IL-7hyFc treatment was comparable to that of GX-188E treatment alone, and the no observed adverse effect level for GX-188E with IL-7hyFc was considered as 320 µg/animal for GX-188E and 1.25 mg/animal for IL-7hyFc.
Assuntos
Fragmentos Fc das Imunoglobulinas/toxicidade , Interleucina-7/toxicidade , Vacinas contra Papillomavirus/toxicidade , Vacinas de DNA/toxicidade , Administração Intravaginal , Animais , Biomarcadores/sangue , Biomarcadores/urina , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Eletroporação , Feminino , Fragmentos Fc das Imunoglobulinas/administração & dosagem , Interleucina-7/administração & dosagem , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Nível de Efeito Adverso não Observado , Vacinas contra Papillomavirus/administração & dosagem , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão/toxicidade , Medição de Risco , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Fatores de Tempo , Vacinas de DNA/administração & dosagemRESUMO
The cell composition of leukocyte infiltrates in the endometrium, myometrium, and vaginal walls was studied in Wistar rats with modeled chronic endomyometritis after administration of IFNγ (0.1 µg/100 g body weight) in different daily regimens (10.00 or 20.00). Morning injections of this cytokine ameliorated inflammatory infiltration of the uterine wall and vagina, but increased the content of neutrophils in the endometrium. Evening cytokine injections reduced neutrophilic infiltration, enhanced mononuclear infiltration, and had no effect on plasmacytic infiltration of the uterine and vaginal walls. In the vaginal wall, both IFNγ administration schedules decreased neutrophil content. The data indicate the necessity to take into account the circadian rhythms in IFN therapy.
Assuntos
Cronofarmacoterapia , Endometrite/tratamento farmacológico , Endométrio/efeitos dos fármacos , Interferon gama/farmacologia , Miométrio/efeitos dos fármacos , Vagina/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Endometrite/imunologia , Endometrite/patologia , Endométrio/imunologia , Endométrio/patologia , Feminino , Humanos , Contagem de Leucócitos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Miométrio/imunologia , Miométrio/patologia , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Plasmócitos/efeitos dos fármacos , Plasmócitos/imunologia , Ratos , Ratos Wistar , Vagina/imunologia , Vagina/patologiaRESUMO
Currently, assessment of the potential immunotoxicity of a given agent involves a tiered approach for hazard identification and mechanistic studies, including observational studies, evaluation of immune function, and measurement of susceptibility to infectious and neoplastic diseases. These studies generally use costly low-throughput mammalian models. Zebrafish, however, offer an excellent alternative due to their rapid development, ease of maintenance, and homology to mammalian immune system function and development. Larval zebrafish also are a convenient model to study the innate immune system with no interference from the adaptive immune system. In this study, a respiratory burst assay (RBA) was utilized to measure reactive oxygen species (ROS) production after developmental xenobiotic exposure. Embryos were exposed to non-teratogenic doses of chemicals and at 96 h post-fertilization, the ability to produce ROS was measured. Using the RBA, 12 compounds with varying immune-suppressive properties were screened. Seven compounds neither suppressed nor enhanced the respiratory burst; five reproducibly suppressed global ROS production, but with varying potencies: benzo[a]pyrene, 17ß-estradiol, lead acetate, methoxychlor, and phenanthrene. These five compounds have all previously been reported as immunosuppressive in mammalian innate immunity assays. To evaluate whether the suppression of ROS by these compounds was a result of decreased immune cell numbers, flow cytometry with transgenic zebrafish larvae was used to count the numbers of neutrophils and macrophages after chemical exposure. With this assay, benzo[a]pyrene was found to be the only chemical that induced a change in the number of immune cells by increasing macrophage but not neutrophil numbers. Taken together, this work demonstrates the utility of zebrafish larvae as a vertebrate model for identifying compounds that impact innate immune function at non-teratogenic levels and validates measuring ROS production and phagocyte numbers as metrics for monitoring how xenobiotic exposure alters the innate immune system.
Assuntos
Benzo(a)pireno/efeitos adversos , Testes Imunológicos de Citotoxicidade/métodos , Imunidade Inata/efeitos dos fármacos , Espécies Reativas de Oxigênio/análise , Explosão Respiratória/efeitos dos fármacos , Animais , Animais Geneticamente Modificados , Contagem de Células Sanguíneas , Embrião não Mamífero , Estradiol/efeitos adversos , Estudos de Viabilidade , Ensaios de Triagem em Larga Escala/métodos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Metoxicloro/efeitos adversos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Compostos Organometálicos/efeitos adversos , Fenantrenos/efeitos adversos , Espécies Reativas de Oxigênio/metabolismo , Explosão Respiratória/imunologia , Peixe-ZebraRESUMO
Mammals have evolved a special cellular mechanism for killing invading microbes, which is called the phagocytosis. Neutrophils are the first phagocytosing cells that migrate into the site of infection. In these cells, hypochlorite (HOCl) and other hypohalites, generated in the myeloperoxidase (MPO)-hydrogen peroxide (H2O2)-halide system is primarily responsible for oxidative killing. Here, we present a method for assessing these oxidative mechanisms in an in vitro cell-free system in a kinetical real-time-based manner by utilizing a bioluminescent bacterial probe called Escherichia coli-lux. The E. coli-lux method provides a practical tool for assessing the effects of various elementary factors in the MPO-H2O2-halide system. Due to the reported versatile intracellular pH and halide concentration during the formation of the phagolysosome and respiratory burst, the antimicrobial activity of the MPO-H2O2-halide system undergoes extensive alterations. Here, we show that at a physiological pH or lower, the antimicrobial activity of MPO is high, and the system effectively enhances the H2O2-dependent oxidative killing of E. coli by chlorination. The HOCl formed in this reaction is a prominent microbe killer. During the respiratory burst, there is a shift to a more alkaline environment. At pH 7.8, the chlorinating activity of MPO was shown to be absent, and the activity of the HOCl decreased. At this higher pH, the activity of H2O2 is enhanced and high enough to kill E. coli without the participation of MPO, and the lowered chloride concentration seemed still to enhance the H2O2-dependent killing capacity.
Assuntos
Anti-Infecciosos/farmacologia , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Oxirredução/efeitos dos fármacos , Peroxidase/metabolismo , Concentração de Íons de Hidrogênio , Ácido Hipocloroso/farmacologia , Neutrófilos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Fagossomos/efeitos dos fármacos , Explosão Respiratória/efeitos dos fármacosRESUMO
BACKGROUND: Glucocorticoids (GC) therapeutic response in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) varies markedly. AIMS/OBJECTIVES: To compare the utility between subjective and objective assessment of GC sensitivity in reflecting the impact of GC on systemic and local eosinophilia in CRSwNP patients. MATERIAL AND METHODS: Twenty-six patients with CRSwNP were enrolled. All patients were given 30 mg of prednisone once daily for 7 days and subsequently classified into subjectively GC-sensitive and -insensitive subgroup or objectively GC-sensitive and -insensitive subgroup. The numbers of eosinophils and neutrophils in blood and polyp tissues were compared between GC-sensitive and GC-insensitive subgroup. RESULTS: 17/26 (65.4%) patients were subjectively and 8/26 (30.8%) patients objectively sensitive to GC treatment. The absolute number and percentage of eosinophils in blood were decreased both in GC-sensitive and -insensitive subjects after GC treatment. In addition, a significant reduction in tissue eosinophil percentage was only observed in objectively GC-sensitive subjects after GC treatment. Furthermore, the change of tissue eosinophil percentage in objectively GC-sensitive subjects was significantly higher than that in objectively GC-insensitive subjects. CONCLUSIONS AND SIGNIFICANCE: Objective assessment may better reflect oral GC response in tissue eosinophilic inflammation than subjective assessment in patients with CRSwNP.
Assuntos
Glucocorticoides/uso terapêutico , Pólipos Nasais/tratamento farmacológico , Prednisona/uso terapêutico , Índice de Gravidade de Doença , Adolescente , Adulto , Eosinofilia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/imunologia , Neutrófilos/efeitos dos fármacos , Projetos Piloto , Estudos Prospectivos , Adulto JovemRESUMO
Individuals of African ancestry in the United States and Europe are at increased risk of developing schizophrenia and have poorer clinical outcomes. The antipsychotic clozapine, the only licensed medication for treatment-resistant schizophrenia, is under-prescribed and has high rates of discontinuation in individuals of African ancestry, due in part to increased rates of neutropenia. The genetic basis of lower neutrophil levels in those of African ancestry has not previously been investigated in the context of clozapine treatment. We sought to identify risk alleles in the first genome-wide association study of neutrophil levels during clozapine treatment, in 552 individuals with treatment-resistant schizophrenia and robustly inferred African genetic ancestry. Two genome-wide significant loci were associated with low neutrophil counts during clozapine treatment. The most significantly associated locus was driven by rs2814778 (ß = -0.9, P = 4.21 × 10-21), a known regulatory variant in the atypical chemokine receptor 1 (ACKR1) gene. Individuals homozygous for the C allele at rs2814778 were significantly more likely to develop neutropenia and have to stop clozapine treatment (OR = 20.4, P = 3.44 × 10-7). This genotype, also termed "Duffy-null", has previously been shown to be associated with lower neutrophil levels in those of African ancestry. Our results indicate the relevance of the rs2814778 genotype for those taking clozapine and its potential as a pharmacogenetic test, dependent on the outcome of additional safety studies, to assist decision making in the initiation and on-going management of clozapine treatment.
Assuntos
Clozapina/efeitos adversos , Neutropenia/induzido quimicamente , Neutropenia/genética , Alelos , Antipsicóticos/uso terapêutico , População Negra/genética , Clozapina/administração & dosagem , Sistema do Grupo Sanguíneo Duffy/genética , Feminino , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla/métodos , Humanos , Masculino , Neutropenia/sangue , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Neutrófilos/patologia , Polimorfismo de Nucleotídeo Único , Receptores de Superfície Celular/genética , Fatores de Risco , Esquizofrenia/genéticaRESUMO
RATIONALE: Cystic fibrosis (CF) lung disease progresses by a combination of airway inflammation, bacterial colonization, and infection. Airway inflammation is predominantly neutrophilic and complicates airway clearance therapies through cellular debris; excessive DNA; excessive and viscous mucus; and high concentrations of neutrophils, IL-8, and related cytokines liberated along the nuclear factor-κB signaling pathway. OBJECTIVES: We conducted a preliminary, single-site, randomized, double-blind, placebo-controlled study to evaluate the effects over 28 days of two dose levels (0.05 mg and 0.1 mg daily) of an older cardiac glycoside, digitoxin, as compared with placebo, on safety, pharmacokinetics, and inflammatory markers in induced sputum obtained from 24 subjects with mild to moderate CF lung disease. METHODS: Patients with CF 18-45 years old with any genotype combination were eligible. The primary objective was to measure the effects of digitoxin on IL-8 and neutrophil counts in induced sputum. Secondary objectives were to measure (1) the pharmacokinetics of digitoxin in sera of patients with stable CF; (2) safety indices, including ECG changes and sputum microbiology; (3) the effect of digitoxin on gene expression in nasal epithelial cells of patients with stable CF; and (4) quality-of-life scores using the Cystic Fibrosis Questionnaire-Revised. MEASUREMENTS AND MAIN RESULTS: It took several weeks to achieve a therapeutic serum level of digitoxin in subjects with CF. No safety concerns emerged during the study. Digitoxin treatment showed a trend toward reduction in sputum free neutrophil elastase and neutrophil counts, but not a reduction in sputum IL-8. Digitoxin treatment did not reach statistical significance for the primary or secondary outcome measures over the 28-day study period. However, the nasal mRNA from the group receiving 0.1 mg of digitoxin daily had a distinct distribution of global gene expression levels as compared with either the 0.05-mg dose or placebo treatment. The mRNAs encoding chemokine/cytokine or cell surface receptors in immune cells were decreased in nasal epithelial cells at the higher dose, leading to pathway-mediated reductions in IL-8, IL-6, lung epithelial inflammation, neutrophil recruitment, and mucus hypersecretion. CONCLUSIONS: At a dose of 0.1 mg daily for 28 days, digitoxin was safe for adults with CF lung disease, but it did not achieve a significant decrease in sputum inflammatory markers. Clinical trial registered with www.clinicaltrials.gov (NCT00782288).
Assuntos
Anti-Inflamatórios/uso terapêutico , Fibrose Cística/tratamento farmacológico , Digitoxina/uso terapêutico , Inflamação/tratamento farmacológico , Adolescente , Adulto , Anti-Inflamatórios/farmacocinética , Biomarcadores/química , Digitoxina/farmacocinética , Método Duplo-Cego , Feminino , Humanos , Inflamação/microbiologia , Interleucina-8/química , Contagem de Leucócitos , Pulmão/fisiopatologia , Masculino , Maryland , Neutrófilos/efeitos dos fármacos , Escarro/química , Escarro/microbiologia , Adulto JovemRESUMO
The present study explores the preparation, characterization and the role of phenolic acid tethered fibrous protein in the management of induced oxidative stress studied under in vitro conditions. In brief, the biomaterial is prepared by engineering the fibrous protein with dihydroxy and trihydroxy phenolic acid moieties and subjected to characterization to ensure the tethering. The resultant biomaterial studied for its efficacy as a free radical scavenger using polymorphonuclear (PMN) cells with induced oxidative stress and also as an agent for cell migration using fibroblasts cells. Results revealed that induced oxidative stress in PMN cells after exposure to UVB radiation managed well with the prepared biomaterial by reducing the levels of superoxide anion, oxygen and hydroxyl radicals. Further, the protein and the phenolic acid interaction supports the cell migration as evidenced from the scratch assay. In conclusion, though phenolic acids are well known for their antimicrobial and antioxidant potential, indenting these acids directly to the wounds is not sensible, but tethering to protein explored the scavenging activity as expected. The present study infers that phenolic acid engineered protein has a significant role in managing the imbalance in the redox state prevailing in wounds and supports the healing at appreciable level.
Assuntos
Hidroxibenzoatos/química , Hidroxibenzoatos/farmacologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Escleroproteínas/química , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Compostos de Bifenilo/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Espaço Extracelular/efeitos dos fármacos , Espaço Extracelular/metabolismo , Espaço Extracelular/efeitos da radiação , Radical Hidroxila/metabolismo , Metaloproteinases da Matriz/metabolismo , Neutrófilos/citologia , Neutrófilos/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Picratos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxidos/metabolismo , Suínos , Raios UltravioletaRESUMO
The bark of A. occidentale L. is rich in tannins. Studies have described various biological activities of the plant, including antimicrobial, antioxidant, antiulcerogenic and antiinflammatory actions. The objective of this study was to assess the activity of the ethyl acetate phase (EtOAc) of A. occidentale on acute inflammation and to identify and quantify its phenolic compounds by HPLC. The method was validated and shown to be linear, precise and accurate for catechin, epicatechin, epigallocatechin and gallic acid. Swiss albino mice (Mus musculus) were treated with saline, Carrageenan (2.5%), Indomethacin (10 mg/kg), Bradykinin (6 nmol) and Prostaglandine E2 (5 µg) at different concentrations of EtOAc - A. occidentale (12.5; 25; 50; and 100 mg/kg/weight p.o.) for the paw edema test. Challenge was performed with carrageenan (500 µg/mL i.p.) for the doses 50 and 100 mg/kg of EtOAc. Levels of cytokines IL-1, TNF-α, IL-6 and IL-10 were also measured. All EtOAc - A. occidentale concentrations reduced the edema. At 50 and 100 mg/kg, an anti-inflammatory response of the EtOAc was observed. Carrageenan stimulus produced a neutrophil count of 28.6% while 50 and 100 mg/kg of the phase reduced this to 14.5% and 9.1%, respectively. The EtOAc extract reduced levels of IL-1 and TNF-α. These results suggest that the EtOAc plays a modulatory role in the inflammatory response. The chromatographic method can be used for the analysis of the phenolic compounds of the EtOAc phase.
Assuntos
Acetatos/química , Anacardium/química , Anti-Inflamatórios/administração & dosagem , Edema/tratamento farmacológico , Fenóis/administração & dosagem , Casca de Planta/química , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Carragenina/efeitos adversos , Cromatografia Líquida de Alta Pressão , Citocinas/metabolismo , Modelos Animais de Doenças , Edema/induzido quimicamente , Edema/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos , Neutrófilos/efeitos dos fármacos , Fenóis/química , Fenóis/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologiaRESUMO
Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are responsible for most mortality in patients with chronic obstructive pulmonary disease (COPD) and are caused mainly by bacterial infection. We analyzed and compared neutrophil CD64 expression (using the ratio of CD64 level in neutrophils to that in lymphocytes as an index), serum C-reactive protein (CRP), procalcitonin (PCT) levels, white blood cell (WBC) count, and neutrophil percentage among healthy subjects and patients with stable COPD or AECOPD. Compared with patients with COPD and healthy subjects, patients with AECOPD demonstrated significantly increased CD64 index, CRP, PCT, WBC count, and neutrophil percentage. Interestingly, CD64 index and PCT were both significantly higher in patients with AECOPD with positive bacterial sputum culture than those with negative culture. Furthermore, CD64 index and PCT were positively correlated in AECOPD, and there was also correlation between CD64 index and CRP, WBC, and neutrophil percentage. These data suggest that CD64 index is a relevant marker of bacterial infection in AECOPD. We divided patients with AECOPD into CD64-guided group and conventional treatment group. In CD64-guided group, clinicians prescribed antibiotics based on CD64 index; while in the conventional treatment group, clinicians relied on experience and clinical symptoms to determine the necessity for antibiotics. We found that the efficacy of antibiotic treatment in CD64-guided group was significantly improved compared with the conventional treatment group, including reduction of hospital stays and cost and shortened antibiotic treatment duration. Thus, the CD64 index has important diagnostic and therapeutic implications for antibiotic treatment of patients with AECOPD.
Assuntos
Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Positivas/imunologia , Linfócitos/imunologia , Neutrófilos/imunologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Aguda , Idoso , Antibacterianos/economia , Antibacterianos/uso terapêutico , Biomarcadores/sangue , Proteína C-Reativa/imunologia , Proteína C-Reativa/metabolismo , Calcitonina/sangue , Calcitonina/imunologia , Estudos de Casos e Controles , Medicina Baseada em Evidências , Feminino , Infecções por Bactérias Gram-Negativas/complicações , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Positivas/complicações , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Tempo de Internação/economia , Contagem de Leucócitos , Linfócitos/efeitos dos fármacos , Linfócitos/microbiologia , Linfócitos/patologia , Masculino , Pessoa de Meia-Idade , Neutrófilos/efeitos dos fármacos , Neutrófilos/microbiologia , Neutrófilos/patologia , Doença Pulmonar Obstrutiva Crônica/complicações , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/microbiologia , Receptores de IgG/sangue , Receptores de IgG/imunologiaRESUMO
OBJECTIVE: Kawasaki disease (KD) is the most commonly encountered inflammatory disease in children. However, its pathogenesis and diagnostic biomarkers have not been fully investigated. We examined the activation of neutrophils and monocytes in KD. METHODS: We studied the expression of the Fcγ-receptors CD64 and CD16 on neutrophils and monocytes in KD before and after the treatment with intravenous infusion of high dose immunoglobulin (IVIG). Bacterial infections were addressed as well. RESULTS: CD64 expression on neutrophils and monocytes was dramatically increased at the onset of KD flare-ups, but later decreased just after IVIG. Similarly, CD16-positive monocytes were observed at the onset and were less apparent after therapy. The addition of immunoglobulin did not block the expression of CD64 or CD16 in vitro. Serum G-CSF in the majority of patients, and IFN-γ in some patients, were elevated during flares but decreased after treatment. CONCLUSION: Our findings demonstrate that remarkable CD64 expression during KD flare-ups may serve as a biomarker for diagnosis. Evaluation of CD64 is also potentially useful for the determination of treatment efficacy in KD.
Assuntos
Monócitos/metabolismo , Síndrome de Linfonodos Mucocutâneos/metabolismo , Neutrófilos/metabolismo , Receptores de IgG/metabolismo , Biomarcadores/metabolismo , Pré-Escolar , Citocinas/sangue , Feminino , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Masculino , Monócitos/efeitos dos fármacos , Síndrome de Linfonodos Mucocutâneos/sangue , Síndrome de Linfonodos Mucocutâneos/diagnóstico , Síndrome de Linfonodos Mucocutâneos/tratamento farmacológico , Neutrófilos/efeitos dos fármacosRESUMO
There are limited data available supporting the use of the recombinant granulocyte colony-stimulating factor (G-CSF), tbo-filgrastim, rather than traditionally used filgrastim to mobilize peripheral blood stem cells (PBSC) or to accelerate engraftment after autologous stem cell transplantation (ASCT). We sought to compare the efficacy and cost of tbo-filgrastim to filgrastim in these settings. Patients diagnosed with lymphoma or plasma cell disorders undergoing G-CSF mobilization, with or without plerixafor, were included in this retrospective analysis. The primary outcome was total collected CD34(+) cells/kg. Secondary mobilization endpoints included peripheral CD34(+) cells/µL on days 4 and 5 of mobilization, adjunctive use of plerixafor, CD34(+) cells/kg collected on day 5, number of collection days and volumes processed, number of collections reaching 5 million CD34(+) cells/kg, and percent reaching target collection goal in 1 day. Secondary engraftment endpoints included time to neutrophil and platelet engraftment, number of blood product transfusions required before engraftment, events of febrile neutropenia, and length of stay. A total of 185 patients were included in the final analysis. Patients receiving filgrastim (n = 86) collected a median of 5.56 × 10(6) CD34(+) cells/kg, compared with a median of 5.85 × 10(6) CD34(+) cells/kg in the tbo-filgrastim group (n = 99; P = .58). There were no statistically significant differences in all secondary endpoints with the exception of apheresis volumes processed (tbo-filgrastim, 17.0 liters versus filgrastim, 19.7 liters; P < .01) and mean platelet transfusions (tbo-filgrastim, 1.7 units versus filgrastim, 1.4 units; P = .04). In conclusion, tbo-filgrastim demonstrated similar CD34(+) yield compared with filgrastim in mobilization and post-transplantation settings, with no clinically meaningful differences in secondary efficacy and safety endpoints. Furthermore, tbo-filgrastim utilization was associated with cost savings of approximately $1406 per patient utilizing average wholesale price.
Assuntos
Filgrastim/farmacologia , Mobilização de Células-Tronco Hematopoéticas/economia , Transplante de Células-Tronco Hematopoéticas , Linfoma de Células B/terapia , Linfoma de Células T/terapia , Mieloma Múltiplo/terapia , Neoplasias de Plasmócitos/terapia , Adulto , Idoso , Antígenos CD34/imunologia , Benzilaminas , Plaquetas/citologia , Plaquetas/efeitos dos fármacos , Plaquetas/imunologia , Análise Custo-Benefício , Ciclamos , Feminino , Filgrastim/análogos & derivados , Filgrastim/economia , Sobrevivência de Enxerto , Mobilização de Células-Tronco Hematopoéticas/métodos , Compostos Heterocíclicos/uso terapêutico , Humanos , Tempo de Internação/economia , Linfoma de Células B/economia , Linfoma de Células B/imunologia , Linfoma de Células B/patologia , Linfoma de Células T/economia , Linfoma de Células T/imunologia , Linfoma de Células T/patologia , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/economia , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/patologia , Neoplasias de Plasmócitos/economia , Neoplasias de Plasmócitos/imunologia , Neoplasias de Plasmócitos/patologia , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Estudos Retrospectivos , Transplante AutólogoRESUMO
Systemic lupus erythematosus (SLE) is a chronic inflammatory disease mainly characterized by B cell hyperactivity. Glucocorticoid (GC) is widely used in SLE for its potent anti-inflammatory and immunosuppressive effects. Despite its important clinical efficacy, high-dose or long-term use of GC can cause severe side effects, such as osteoporosis, osteonecrosis, cataracts, hyperglycemia, coronary heart disease and cognitive impairment. Our early clinical studies have shown that Jieduquyuzishen prescription (JP) can effectively reduce the adverse effects and improve the curative effect of GC in the treatment of SLE. The BAFF/BAFF-R signaling pathway plays an important role in the development of SLE and has been regarded as a potential target for the therapy of SLE. In this study, we attempt to investigate the effect of JP on the BAFF/BAFF-R signaling pathway to explore the mechanism of JP in reducing the toxicity and enhancing the efficacy of GC. YAC-1 cells, isolated rat peripheral blood lymphocytes, polymorphonuclear neutrophils and spleen lymphocytes were treated with drug-containing serum. The results of RT-PCR, Western blot and dual-luciferase reporter gene assays indicate that either JP or GC can inhibit the mBAFF-induced up-regulation of BAFF, BAFF-R, Bcl-2, IL-10 and NF-κB in YAC-1 cells and WEHI-231 cells. Furthermore, MTS, flow cytometry and CFSE results reveal that the proliferation and survival of lymphocytes activated by mBAFF are suppressed by JP, GC and their combination. Contrary to GC, JP can reduce the apoptosis and raise the survival of polymorphonuclear neutrophils and can't increase the apoptosis of the peripheral blood lymphocytes and spleen lymphocytes. Therefore, it is possible that JP can down-regulate the BAFF/BAFF-R signaling pathway as effectively as GC, which may result in the dosage reduction of GC, thus decreasing the toxicity and improving the efficacy of GC-based treatment of SLE.
Assuntos
Fator Ativador de Células B/metabolismo , Receptor do Fator Ativador de Células B/metabolismo , Prescrições de Medicamentos , Medicamentos de Ervas Chinesas/farmacologia , Soro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Fator Ativador de Células B/genética , Receptor do Fator Ativador de Células B/genética , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sinergismo Farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Glucocorticoides/efeitos adversos , Glucocorticoides/farmacologia , Interleucina-10/genética , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Lúpus Eritematoso Sistêmico/metabolismo , Lúpus Eritematoso Sistêmico/patologia , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Fosfoproteínas/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Baço/imunologia , Transcrição Gênica/efeitos dos fármacosRESUMO
This work evaluated the physicochemical properties and antioxidant activity of spray dried extracts (SDE) from Psidium guajava L. leaves. Different drying carriers, namely, maltodextrin, colloidal silicon dioxide, Arabic gum, and ß -cyclodextrin at concentrations of 40 and 80% relative to solids content, were added to drying composition. SDE were characterized through determination of the total phenolic, tannins, and flavonoid content. Antioxidant potential of the SDE was assessed by two assays: cellular test that measures the luminol-enhanced chemiluminescence (LumCL) produced by neutrophils stimulated with phorbol myristate acetate (PMA) and the DPPH radical scavenging (DPPH∗ method). In both assays the antioxidant activity of the SDE occurred in a concentration-dependent manner and showed no toxicity to the cells. Using the CLlum method, the IC50 ranged from 5.42 to 6.50 µg/mL. The IC50 of the SDE ranged from 7.96 to 8.11 µg/mL using the DPPH(â¢) method. Psidium guajava SDE presented significant antioxidant activity; thus they show high potential as an active phytopharmaceutical ingredient. Our findings in human neutrophils are pharmacologically relevant since they indicate that P. guajava SDE is a potential antioxidant and anti-inflammatory agent in human cells.
Assuntos
Antioxidantes/farmacologia , Neutrófilos/efeitos dos fármacos , Extratos Vegetais/química , Folhas de Planta/química , Psidium/química , Adolescente , Adulto , Antioxidantes/química , Compostos de Bifenilo/análise , Compostos de Bifenilo/química , Compostos de Bifenilo/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Humanos , Medições Luminescentes , Masculino , Neutrófilos/metabolismo , Picratos/análise , Picratos/química , Picratos/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Adulto JovemRESUMO
INTRODUCTION: A pegylated recombinant human granulocyte colony-stimulating factor (G-CSF) is effective in reducing the severity and duration of neutropenia. This study was performed to investigate the pharmacokinetics (PK), pharmacodynamics (PD), and tolerability of GCPGC, a new formulation of pegylated G-CSF, in healthy volunteers and to compare them with those of pegfilgrastim (Neulasta®). METHODS: Twenty-five healthy Korean male volunteers randomly received a single subcutaneous (SC) GCPGC injection at a dose of 30 (n = 10), 100 (n = 10), or 300 (n = 5) µg/kg or placebo in a 4:1 ratio in a double-blind manner. Additionally, 8 subjects received a SC dose of pegfilgrastim at 100 µg/kg. Blood samples were collected up to 14 days after both therapies. The absolute neutrophil count (ANC) and CD34(+) cell counts were the PD markers. RESULTS: After GCPGC administration, 4 different pharmacokinetic phases were identified, indicating target-mediated drug disposition (TMDD) for the elimination of GCPGC, which was slowed down as the dose was increased, resulting in a higher than proportional dose-normalized exposure to GCPGC. Although GCPGC was cleared faster than pegfilgrastim, leading to a 19 % lower systemic exposure to pegylated G-CSF, the increase in ANC and CD34(+) were ~20 % greater by GCPGC at 100 µg/kg than pegfilgrastim. Thrombocytopenia, splenomegaly, and hemoperitoneum occurred in one subject in the 300 µg/kg GCPGC group, which resolved completely with appropriate care. CONCLUSIONS: GCPGC showed a non-linear TMDD. The PK-PD characteristics of GCPGC at 30-100 µg/kg were comparable to those of pegfilgrastim at 100 µg/kg. GCPGC at 30-100 µg/kg was well tolerated in healthy Korean males.
Assuntos
Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos/farmacocinética , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/farmacocinética , Adulto , Antígenos CD34/metabolismo , Relação Dose-Resposta a Droga , Método Duplo-Cego , Filgrastim , Fator Estimulador de Colônias de Granulócitos/efeitos adversos , Voluntários Saudáveis , Humanos , Injeções Subcutâneas/métodos , Contagem de Leucócitos/métodos , Masculino , Neutropenia/tratamento farmacológico , Neutrófilos/efeitos dos fármacos , Polietilenoglicóis/efeitos adversos , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/farmacocinética , Adulto JovemRESUMO
The problem of power and sample size determination for distribution-free multiple comparison tests of K treatments versus a control group is addressed. We define the power as the probability of correctly rejecting one specified or all K hypotheses, corresponding to the per-pair and all-pairs power, respectively. The power formulas are derived for both joint ranking and pairwise ranking mechanism for general multiple comparison problems, followed by explicit form of these formulas when the single-step, step-down, or step-up adjustments are applied. The proposed power and sample size calculation methods apply to scenarios both when the underlying distributions are known and when they are unknown but a pilot study is available. Numerical methods via quasi-Monte Carlo integration and Monte Carlo integration are assessed. Our simulation studies show the accuracy of the power and sample size calculation formulas. We recommend the Monte Carlo integration as the calculation algorithm. An example from a mouse peritoneal cavity study is used to demonstrate the application of the methods.
Assuntos
Bioensaio/estatística & dados numéricos , Interpretação Estatística de Dados , Modelos Estatísticos , Tamanho da Amostra , Algoritmos , Animais , Anti-Inflamatórios/farmacologia , Simulação por Computador , Camundongos , Modelos Animais , Método de Monte Carlo , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Análise Numérica Assistida por Computador , Cavidade Peritoneal , Resultado do TratamentoRESUMO
BACKGROUND: Adhesion formation after abdominal surgery is a major cause of postoperative bowel obstruction, infertility, and chronic abdominal pain. In this study, we evaluated the effect of normal saline and methylene blue (MB) on postoperative adhesion formation in a rat cecum model. METHODS: A total of 30 Wistar female rats in 2 treatment and 1 control groups underwent midline laparotomy and standardized abrasion of the visceral peritoneum. Normal saline and methylene blue were administrated intraperitoneally at the end of the surgical procedure in 2 treatment groups. Fourteen days after surgery, a re-laparotomy was performed for macroscopic and pathological assessment. RESULTS: The adhesion grade and extent of the normal saline group was lower than control and MB groups in macroscopic assessment (P<0.05 for both). A comparison of adhesion stages in pathological assessment showed increment in abdominal adhesion by usage methylene blue 1% and demonstrated significant difference between MB and 2 other groups (P<0.05). CONCLUSIONS: Administrated normal saline individually reduce the adhesion grade near cecum. Conversely, usage of methylene blue 1% may unpredictably increase risk of adhesion formation.