RESUMO
Controlling RAS mutant cancer progression remains a significant challenge in developing anticancer drugs. Whereas Ras G12C-covalent binders have received clinical approval, the emergence of further mutations, along with the activation of Ras-related proteins and signals, has led to resistance to Ras binders. To discover novel compounds to overcome this bottleneck, we focused on the concurrent and sustained blocking of two major signaling pathways downstream of Ras. To this end, we synthesized 25 drug-drug conjugates (DDCs) by combining the MEK inhibitor trametinib with Akt inhibitors using seven types of linkers with structural diversity. The DDCs were evaluated for their cell permeability/accumulation and ability to inhibit proliferation in RAS-mutant cell lines. A representative DDC was further evaluated for its effects on signaling proteins, induction of apoptosis-related proteins, and the stability of hepatic metabolic enzymes. These in vitro studies identified a series of DDCs, especially those containing a furan-based linker, with promising properties as agents for treating RAS-mutant cancers. Additionally, in vivo experiments in mice using the two selected DDCs revealed prolonged half-lives and anticancer efficacies comparable to those of trametinib. The PK profiles of trametinib and the Akt inhibitor were unified through the DDC formation. The DDCs developed in this study have potential as drug candidates for the broad inhibition of RAS-mutant cancers.
Assuntos
Antineoplásicos , Neoplasias , Animais , Camundongos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias/tratamento farmacológico , Neoplasias/genética , Transdução de Sinais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Mutação , Inibidores da Angiogênese/farmacologia , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Linhagem Celular TumoralRESUMO
BACKGROUND: Although Korea and Japan have a national gastric cancer screening program, their screening intervals are different. The optimal screening interval of endoscopic screening in Japan was investigated based on the stage distributions of screen-detected gastric cancers. METHODS: Patients with gastric cancer detected by endoscopic and radiographic screenings were selected from the Niigata City Medical Association database. The stage distributions of the detected gastric cancers were compared among patients with different screening histories in both groups. Gastric cancer specific survival rates were analyzed using the Kaplan-Meier method with the log-rank test. RESULTS: There were 1585 and 462 subjects in the endoscopic and radiographic screening groups, respectively. In the endoscopic screening group, the stage IV proportion was lower in patients with screening history 1 and 2 years before diagnosis than in patients without screening history. Stage IV development was significantly related to the absence of screening history (p < 0.001); however, there were no differences between patients who had endoscopic screening history 2 and 3 years before diagnosis. The survival rates were not significantly different between patients with endoscopic screening 1 and 2 years previously (p = 0.7763). The survival rates were significantly higher in patients with endoscopic screening history 1 and 2 years before diagnosis than in patients without screening history (p < 0.001), and in patients with endoscopic screaming 3 years before diagnosis (P < 0.0069). CONCLUSION: The endoscopic screening interval for gastric cancer can be expanded to at least 2 years based on the stage distributions of detected cancers and the patient survival rates.
Assuntos
Detecção Precoce de Câncer/normas , Endoscopia Gastrointestinal/normas , Neoplasias Gástricas/diagnóstico , Adulto , Idoso , Detecção Precoce de Câncer/métodos , Detecção Precoce de Câncer/mortalidade , Endoscopia Gastrointestinal/métodos , Feminino , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias/mortalidade , Neoplasias Gástricas/mortalidade , Taxa de Sobrevida/tendências , Fatores de TempoRESUMO
BACKGROUND: There is no systematic analysis to identify problems involved with instruction on inhalation therapy for elderly patients. We conducted a nationwide questionnaire survey for patients and medical professionals. METHODS: A questionnaire survey was conducted of adult patients on inhaled drugs (ages 18-92 years, 820 individuals) and medical professionals (pharmacists or nurses) who provided instruction on inhalation therapy to these patients in 23 institutions in Japan to investigate the technique and the level of understanding (knowledge) of the inhalation therapy. Changes in the recognition of performance of inhalation technique and inhalation knowledge with increasing age were analyzed. RESULTS: According to patients' subjective assessment, there was no deterioration in the performance of the inhalation technique or loss of the knowledge with increasing age. On the other hand, medical professionals' objective assessment revealed a significant loss of both inhalation technique and knowledge with increasing age. Not many elderly patients noticed their own problems themselves, revealing a great perception gap between elderly patients and medical professionals. Thus, there was concern that patients would unconsciously practice the inhalation procedure improperly. On the other hand, in comparison with non-elderly patients, elderly patients were less resistant to continuation of therapy, suggesting that they would be more likely to accept instruction on inhalation therapy. CONCLUSIONS: Elderly patients are apt to assume that they "understand well", therefore, in order to recognize and close the perception gap between elderly patients and medical professionals, it is necessary to provide them with more aggressive (frequent) instructions on inhalation therapy.
Assuntos
Pessoal de Saúde , Pacientes , Terapia Respiratória/estatística & dados numéricos , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Feminino , Avaliação Geriátrica , Pessoal de Saúde/estatística & dados numéricos , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Pacientes/estatística & dados numéricos , Vigilância da População , Curva ROC , Terapia Respiratória/métodos , Inquéritos e Questionários , Adulto JovemRESUMO
Water movement between cells in a plant body is the basic phenomenon of plant solute transport; however, it has not been well documented due to limitations in observational techniques. This paper reports a visualization technique to observe water movement among plant cells in different tissues using a time of flight-secondary ion mass spectrometry (Tof-SIMS) cryo-system. The specific purpose of this study is to examine the route of water supply from xylem to stem tissues. The maximum resolution of Tof-SIMS imaging was 1.8 µm (defined as the three pixel step length), which allowed detection of water movement at the cellular level. Deuterium-labelled water was found in xylem vessels in the stem 2.5 min after the uptake of labelled water by soybean plants. The water moved from the xylem to the phloem, cambium, and cortex tissues within 30-60 min after water absorption. Deuterium ion counts in the phloem complex were slightly higher than those in the cortex and cambium tissue seen in enlarged images of stem cell tissue during high transpiration. However, deuterium ion counts in the phloem were lower than those in the cambium at night with no evaporative demand. These results indicate that the stem tissues do not receive water directly from the xylem, but rather from the phloem, during high evaporative demand. In contrast, xylem water would be directly supplied to the growing sink during the night without evaporative demand.
Assuntos
Glycine max/metabolismo , Água/metabolismo , Xilema/metabolismo , Criopreservação , Espectrometria de Massa de Íon SecundárioRESUMO
Pulmonary alveolar proteinosis (PAP) is a devastating lung disease caused by abnormal surfactant homeostasis, with a prevalence of 6-7 cases per million population worldwide. While mutations causing hereditary PAP have been reported, the genetic basis contributing to autoimmune PAP (aPAP) has not been thoroughly investigated. Here, we conducted a genome-wide association study of aPAP in 198 patients and 395 control participants of Japanese ancestry. The common genetic variant, rs138024423 at 6p21, in the major-histocompatibility-complex (MHC) region was significantly associated with disease risk (Odds ratio [OR] = 5.2; P = 2.4 × 10-12). HLA fine-mapping revealed that the common HLA class II allele, HLA-DRB1*08:03, strongly drove this signal (OR = 4.8; P = 4.8 × 10-12), followed by an additional independent risk allele at HLA-DPß1 amino acid position 8 (OR = 0.28; P = 3.4 × 10-7). HLA-DRB1*08:03 was also associated with an increased level of anti-GM-CSF antibody, a key driver of the disease (ß = 0.32; P = 0.035). Our study demonstrated a heritable component of aPAP, suggesting an underlying genetic predisposition toward an abnormal antibody production.
Assuntos
Autoanticorpos/genética , Doenças Autoimunes/genética , Predisposição Genética para Doença , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Cadeias HLA-DRB1/genética , Proteinose Alveolar Pulmonar/genética , Adulto , Idoso , Alelos , Povo Asiático , Autoanticorpos/biossíntese , Doenças Autoimunes/etnologia , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Estudos de Casos e Controles , Cromossomos Humanos Par 6 , Feminino , Expressão Gênica , Frequência do Gene , Estudo de Associação Genômica Ampla , Fator Estimulador de Colônias de Granulócitos e Macrófagos/antagonistas & inibidores , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Cadeias HLA-DRB1/imunologia , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Razão de Chances , Isoformas de Proteínas/genética , Proteinose Alveolar Pulmonar/etnologia , Proteinose Alveolar Pulmonar/imunologia , Proteinose Alveolar Pulmonar/patologia , Surfactantes Pulmonares/imunologia , Surfactantes Pulmonares/metabolismo , RiscoRESUMO
A 70-year-old man with gastric cancer of Borrmann type 3, liver metastases and peritoneal dissemination was treated by combination therapy of S-1 and docetaxel (DOC). He received DOC intravenously at 40 mg/m(2) on day 1 and S-1 orally at 100 mg/body on day 1 to 14, repeated every 28 days. After 2 courses of treatment, a CT scan revealed improvement of the gastric wall thickness, the eminent decrease of the peritoneal fluid and the reduction of the liver metastasis. After 3 courses of treatment, the primary lesion was remarkably improved on endoscopic examination, and the tumor marker normalized after 4 courses of treatment. Toxicities included leukocytopenia (WHO grade 3), neutropenia ( grade 3), anorexia (grade 2), and nausea (grade 2). Outpatient chemotherapy was possible by reduction of dose (S-1 100--> 80 mg/body, DOC 40--> 32 mg/m2). The response was maintained on CT and endoscopic examination after 21 courses of treatment. A case of an advanced gastric cancer patient successfully treated by combination therapy of S-1 and DOC was reported.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Ácido Oxônico/uso terapêutico , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/patologia , Taxoides/uso terapêutico , Tegafur/uso terapêutico , Idoso , Biomarcadores Tumorais/sangue , Docetaxel , Combinação de Medicamentos , Feminino , Gastroscopia , Humanos , Masculino , Estadiamento de Neoplasias , Neoplasias Gástricas/sangue , Tomografia Computadorizada por Raios XRESUMO
Amenamevir is a potent helicase-primase inhibitor and a novel class of antiviral agent other than nucleoside compounds, such as aciclovir, valaciclovir and famciclovir. This study is the first randomized, double-blind, valaciclovir-controlled phase 3 study to evaluate the efficacy and safety of amenamevir in Japanese patients with herpes zoster when treated within 72 h after onset of rash. A total of 751 patients were randomly assigned to receive either amenamevir 400 mg or 200 mg p.o. once daily or valaciclovir 1000 mg three times daily (daily dose, 3000 mg) for 7 days. The primary efficacy end-point was the proportion of cessation of new lesion formation by day 4 ("day 4 cessation proportion"). The day 4 cessation proportions for amenamevir 400 and 200 mg and valaciclovir were 81.1% (197/243), 69.6% (172/247) and 75.1% (184/245), respectively. Non-inferiority of amenamevir 400 mg to valaciclovir was confirmed by a closed testing procedure. Days to cessation of new lesion formation, complete crusting, healing, pain resolution and virus disappearance were evaluated as secondary end-points. No significant differences were observed in any of the treatment groups. Amenamevir 400 and 200 mg were well tolerated as well as valaciclovir. The proportions of patients who experienced drug-related adverse events were 10.0% (25/249), 10.7% (27/252) and 12.0% (30/249) with amenamevir 400 and 200 mg and valaciclovir, respectively. In conclusion, amenamevir 400 mg appears to be effective and well tolerated for treatment of herpes zoster in immunocompetent Japanese patients.
Assuntos
Antivirais/administração & dosagem , Herpes Zoster/tratamento farmacológico , Oxidiazóis/administração & dosagem , Aciclovir/análogos & derivados , Aciclovir/uso terapêutico , Idoso , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Valaciclovir , Valina/análogos & derivados , Valina/uso terapêuticoRESUMO
An image super-resolution method from multiple observation of low-resolution images is proposed. The method is based on sub-pixel accuracy block matching for estimating relative displacements of observed images, and sparse signal representation for estimating the corresponding high-resolution image, where correspondence between high- and low-resolution images are modeled by a certain degradation process. Relative displacements of small patches of observed low-resolution images are accurately estimated by a computationally efficient block matching method. The matching scores of the block matching are used to select a subset of low-resolution patches for reconstructing a high-resolution patch, that is, an adaptive selection of informative low-resolution images is realized. The proposed method is shown to perform comparable or superior to conventional super-resolution methods through experiments using various images.
Assuntos
Algoritmos , Aumento da Imagem/métodos , Reconhecimento Automatizado de Padrão/métodosRESUMO
AIM: To investigate mortality reduction from gastric cancer based on the results of endoscopic screening. METHODS: The study population consisted of participants of gastric cancer screening by endoscopy, regular radiography, and photofluorography at Niigata city in 2005. The observed numbers of cumulative deaths from gastric cancers and other cancers were accumulated by linkage with the Niigata Prefectural Cancer Registry. The standardized mortality ratio (SMR) of gastric cancer and other cancer deaths in each screening group was calculated by applying the mortality rate of the reference population. RESULTS: Based on the results calculated from the mortality rate of the population of Niigata city, the SMRs of gastric cancer death were 0.43 (95%CI: 0.30-0.57) for the endoscopic screening group, 0.68 (95%CI: 0.55-0.79) for the regular radiographic screening group, and 0.85 (95%CI: 0.71-0.94) for the photofluorography screening group. The mortality reduction from gastric cancer was higher in the endoscopic screening group than in the regular radiographic screening group despite the nearly equal mortality rates of all cancers except gastric cancer. CONCLUSION: The 57% mortality reduction from gastric cancer might indicate the effectiveness of endoscopic screening for gastric cancer. Further studies and prudent interpretation of results are needed.
Assuntos
Detecção Precoce de Câncer/métodos , Gastroscopia , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Adulto , Idoso , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Radiografia , Sistema de Registros , Neoplasias Gástricas/diagnóstico por imagem , Neoplasias Gástricas/prevenção & controle , Fatores de TempoRESUMO
This paper examined the route of water supply into soybean nodules through the new visualization technique of time of flight secondary ion mass spectrometry (Tof-SIMS) cryo system, and obtained circumstantial evidence for the water inflow route. The maximum resolution of the Tof-SIMS imaging used by this study was 1.8 µm (defined as the three pixel step length), which allowed us to detect water movement at the cellular level. Deuterium-labeled water was supplied to soybean plants for 4h and the presence of deuterium in soybean nodules was analyzed by the Tof-SIMS cryo system. Deuterium ions were found only in the endodermis tissue surrounding the central cylinder in soybean nodules. Neither xylem vessels nor phloem complex itself did not indicate any deuterium accumulation. Deuterium-ion counts in the endodermis tissue were not changed by girdling treatment, which restricted water movement through the phloem complex. The results strongly indicated that nodule tissues did not receive water directly from the phloem complex, but received water through root cortex apoplastic pathway from the root axis.
Assuntos
Glycine max/metabolismo , Água/metabolismo , Transporte Biológico , Deutério/metabolismo , Floema/metabolismo , Nódulos Radiculares de Plantas/metabolismo , Espectrometria de Massa de Íon Secundário , Xilema/metabolismoRESUMO
Human histamine H1 receptor (H1R) contains five possible phosphorylation residues (Thr140, Thr142, Ser396, Ser398 and Thr478) and the substitution of all these five residues to alanine completely impairs agonist-induced receptor downregulation. In the present study, to determine which residue(s) are responsible for receptor downregulation, we used mutant H1Rs in which single or multiple residues were substituted with alanine. The results suggested that two groups, i.e., residues Thr140 and Thr142, and residues Ser396 and Ser398, independently contributed to H1R downregulation. Thr140 and Ser398 mainly contributed to downregulation, and Thr142 or Ser396 had a slight inhibitory effect on Thr140- or Ser398-mediated process, respectively.
Assuntos
Regulação para Baixo , Receptores Histamínicos H1/química , Receptores Histamínicos H1/metabolismo , Serina/metabolismo , Treonina/metabolismo , Sequência de Aminoácidos , Animais , Células CHO , Cricetinae , Regulação para Baixo/efeitos dos fármacos , Histamina/farmacologia , Humanos , Dados de Sequência Molecular , Mutação/genética , Conformação Proteica , Receptores Histamínicos H1/genética , Serina/genética , Treonina/genéticaRESUMO
We report a case of advanced gastric cancer that responded to docetaxel with low-dose 5-FU and cisplatin combination chemotherapy after becoming chemoresistant to M-FLP. A 52-year-old male was diagnosed with type 3 gastric cancer of angulus (poorly differentiated adenocarcinoma) with left neck, Virchow, mediastinal and abdominal lymph nodes metastases. The patient was treated with 5 courses of M-FLP (MTX + 5-FU + LV + CDDP), and the effect of this therapy was PR, but the tumor was chemoresistant to the sixth course of this therapy. After 7 courses of M-FLP, docetaxel (TXT) with low-dose FP (5-FU + CDDP) was administered to the patient as second-line chemotherapy. After 2 courses of TXT with low-dose FP, the gastric cancer and metastatic lymph nodes were remarkably reduced and the effect of this therapy was PR. The toxic events were anemia (grade 2) and leukopenia (grade 3), which were treated with G-CSF. CDDP and 5-FU based regimens are considered as the first-line chemotherapy for metastatic advanced gastric cancer in Japan; however, a second-line chemotherapy has not been established. As in this case, a TXT based regimen is effective and well tolerated therapy as a second-line chemotherapy for metastatic gastric cancer after prior exposure to CDDP and 5-FU.
Assuntos
Adenocarcinoma/tratamento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Neoplasias Gástricas/tratamento farmacológico , Adenocarcinoma/secundário , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Cisplatino/administração & dosagem , Docetaxel , Esquema de Medicação , Fluoruracila/administração & dosagem , Humanos , Leucovorina/administração & dosagem , Metástase Linfática , Masculino , Metotrexato/administração & dosagem , Pessoa de Meia-Idade , Neoplasias Gástricas/patologia , Taxoides/administração & dosagemRESUMO
Graft-versus-host disease(GVHD) colitis after allogeneic bone marrow transplantation or peripheral blood stem cell transplantation was often accompanied with upper gastro-intestinal symptoms such as upper abdominal pain. We investigated 7 cases of gastro-duodenal lesions with GVHD colitis. Endoscopic features of gastric lesions with GVHD were turbidity(100%), erythema(100%), erosions(85.7%), easy bleeding of mucosa(71.4%) and duodenal involvement(71.4%), histological findings of those were infiltration of inflammatory cells(100%), dilatation of capillaries(100%), hemorrhage(71.4%), stromal edema(85.7%), degenerated glands and apoptotic bodies(100%). Those findings resembled with endoscopic and histological features of GVHD colitis, so we diagnosed those lesions as GVHD gastro-duodenitis. Clinically GVHD colitis with gastro-duodenitis was severe compared with colitis without gastro-duodenitis.
Assuntos
Doença Enxerto-Hospedeiro/complicações , Úlcera Péptica/etiologia , Colite/etiologia , Humanos , Úlcera Péptica/patologiaRESUMO
Patients who have undergone distal gastrectomy for peptic ulcer are at higher risk of developing gastric remnant cancer, and chronic bile reflux is believed to increase the risk of cancer in remnant stomach. In remnant stomach, carcinogenesis may be prevented by selecting the anastomosis method with a few reflux of intestinal juice including a bile acid. How Helicobacter pylori(H. pylori) infection participate in stomal gastritis and gastric remnant cancer, same as early gastric cancer in the intact stomach, is attended. H. pylori positive rate of remnant stomach is different by examination method and a report, but its rate is decreased every year after gastrectomy and in particular low in Billroth-II(B-II) anastomosis. B-II anastomosis is followed by a significantly lower rate than B-1. This may reflect the role of bile reflux because bile reflux interferes with colonization by H. pylori. Gastric cancer excision usual increase complicates gastric remnant stomach and H. pylori infection, but while H. pylori infection lasts after gastrectomy for gastric cancer, cell proliferation increase in remnant stomach. In remnant stomach after gastrectomy for gastric cancer, while H. pylori infection continues, H. pylori infection may cause remnant gastritis and a second cancer of remnant stomach. H. pylori infection and bile reflux seem to have a synergistic effect on cell proliferation in remnant stomach and may explain the increased risk of gastric remnant cancer. The cancer-causing dominant role might changed from H. pylori infection predominance to bile reflux every year after gastrectomy. Furthermore, a prophylactic effect to carcinogenesis by H. pylori eradication therapy is expected. Eradication of H. pylori after gastrectomy for gastric cancer has been recommended.
Assuntos
Coto Gástrico , Infecções por Helicobacter/complicações , Helicobacter pylori , Neoplasias Gástricas/etiologia , Gastrectomia , Infecções por Helicobacter/tratamento farmacológico , Humanos , Neoplasias Gástricas/prevenção & controleRESUMO
We herein present the case of a 56-year-old Japanese woman who developed purulent pericarditis after undergoing chemoradiotherapy for esophageal cancer. She developed epigastralgia and a fever and was admitted to our hospital. A physical examination revealed hypotension, tachycardia and pericardial friction rub. Echocardiography revealed moderate pericardial effusion. Based on these observations, the patient was diagnosed with cardiac tamponade. Computed tomography confirmed the presence of an esophagopericardial fistula. Treatment with pericardiocentesis, drainage and short-term intrapericardial administration of antibiotics relieved the patient's symptoms. Daily rinsing through a catheter with normal saline prevented relapse of the purulent pericarditis. Esophagopericardial fistulas are so rare that their treatment is not well-established. We herein report successful palliative care of a malignant esophagopericardial fistula associated with purulent pericarditis.
Assuntos
Fístula Esofágica/diagnóstico , Neoplasias Esofágicas/diagnóstico , Derrame Pericárdico/diagnóstico , Pericardite/diagnóstico , Fístula Esofágica/complicações , Neoplasias Esofágicas/complicações , Feminino , Fístula/complicações , Fístula/diagnóstico , Humanos , Pessoa de Meia-Idade , Derrame Pericárdico/complicações , Pericardite/complicações , Pericárdio/microbiologia , Pericárdio/patologiaAssuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Camptotecina/análogos & derivados , Colo Sigmoide/cirurgia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/secundário , Neoplasias do Colo Sigmoide/patologia , Camptotecina/administração & dosagem , Esquema de Medicação , Combinação de Medicamentos , Fluoruracila/administração & dosagem , Humanos , Irinotecano , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Neoplasias do Colo Sigmoide/cirurgia , Tegafur/administração & dosagem , Uracila/administração & dosagemRESUMO
A convenient method for measuring immune complexes between tissue-nonspecific alkaline phosphatase (TNSALP) and immunoglobulin G (IgG) (i.e., TNSALP-IgG) would be highly useful for routine analyses. Here, we identified a surface-active agent that would dissolve membrane but not dissociate TNSALP-IgG complexes. Next, we developed an enzyme-linked immunosorbent assay (ELISA) method for detecting TNSALP-IgG complexes with two monoclonal antibodies (MoAbs): 3-29-3R was coated on assay plates and captured TNSALP-IgG from a specimen; an horseradish peroxidase (HRP)-conjugated anti-human IgG1 then reacted with captured TNSALP-IgG to form an "immunocomplex sandwich." The immunocomplex was detected via the absorbance of an HRP substrate, resulting in a semiquantitative assay. The mean absorbance of 0.195 (n=5) measured in sera from healthy donors was designated as an arbitrary unit (AU/mL) of TNSALP-IgG concentration. The ELISA values of patient sera known to contain TNSALP-IgG complexes were greater than those of normal sera (normal, 1.86 plusmn; 0.61; patient, 9.30 plusmn; 5.44), and these data were confirmed by electrophoresis. Thus, the ELISA could detect TNSALP-IgG complexes. The intraassay coefficient of variation (CV) was within 7.4% and analytical recovery was excellent. There was no significant interference from hemolytic, lipemic, or icteric serum. In summary, an ELISA using 3-29-3R MoAb and HRP-conjugated anti-human IgG1 constitutes a reliable and convenient method for the semiquantitative detection of TNSALP-IgG complexes in human serum.
Assuntos
Fosfatase Alcalina/imunologia , Complexo Antígeno-Anticorpo/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G/sangue , Fosfatase Alcalina/sangue , Fosfatase Alcalina/metabolismo , Anticorpos Monoclonais , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Eletroforese , Humanos , Imunoglobulina G/metabolismo , Tensoativos/química , Tensoativos/farmacologiaRESUMO
The chemical structure of lignin, a complex, irregular polymer of phenylpropane units that occurs in plant cell walls, was investigated using time-of-flight secondary ion mass spectrometry (ToF-SIMS). The positive ToF-SIMS spectra of lignin isolated from pine and beech wood showed prominent secondary ions possessing guaiacyl (at m/z 137 and 151) or syringyl (at m/z 167 and 181) rings, which are the basic building units of lignin polymer. This shows that ToF-SIMS is a useful tool for lignin structural analysis. The peaks at m/z 137 and 167 were assigned as the C6-C1 ion, and the peaks at m/z 151 and 181 may be double-component, the C6-C1 ion and the C6-C2 ion. We confirmed the characteristic guaiacyl ions using a synthetic lignin model compound, dehydrogenation polymer (DHP), which was formed by polymerizing of unlabeled and deuterium-labeled coniferyl alcohols. The formation mechanism of the main secondary ions was deduced by labeling specific positions of coniferyl alcohols with a stable isotope to study the relationship between chemical structure and secondary ion formation in ToF-SIMS.