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Gastroesophageal reflux disease (GERD) is among the most prevalent gastrointestinal (GI) disorders. It is known to often coexist with other chronic diseases such as asthma, chronic obstructive pulmonary disease (COPD), obesity, diabetes mellitus (DM), and hypertension. Upper endoscopy, esophageal manometry, and impedance-pH monitoring are a few invasive diagnostic options that are reserved for selected GERD patients. Symptom assessment by using questionnaires, such as the frequency scale for the symptoms of GERD (FSSG), is simple, convenient, noninvasive, and inexpensive. These questionnaires are widely used to facilitate diagnosis and appropriate treatment. Early diagnosis of GERD and timely management may improve clinical outcomes in patients. Proton pump inhibitors (PPIs) are the preferred therapy for GERD. However, evidence indicates that excessive and extended use of PPIs is linked to adverse events. An overview of the diagnosis and management of GERD, as well as an evidence-based overview of the relationship between GERD and asthma, COPD, obesity, DM, and hypertension, is presented in this review. Expert opinions and recommendations for diagnosing GERD using invasive tests and validated questionnaires have also been mentioned.
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Asma , Refluxo Gastroesofágico , Hipertensão , Doença Pulmonar Obstrutiva Crônica , Humanos , Refluxo Gastroesofágico/complicações , Refluxo Gastroesofágico/diagnóstico , Refluxo Gastroesofágico/epidemiologia , Obesidade/complicações , Obesidade/diagnóstico , Obesidade/epidemiologia , Inibidores da Bomba de Prótons/uso terapêuticoRESUMO
Determining the adverse nature of findings from nonclinical safety studies often poses a challenge for the key stakeholders responsible for interpreting the results of definitive toxicity studies in support of pharmaceutical product development. Although there are instances in which responses to treatment clearly indicate intolerability or tissue injury associated with dysfunction; in practice, more often there is uncertainty in characterizing an effect of drug treatment as adverse or not. This is due to the inherent variability in responses of biological test systems to toxicological insults, leaving the ultimate analyses of adversity to individual interpretation and subjectivity. This article is a follow-up to the workshop entitled, "Adverse or Not Adverse?: Thinking process behind adversity determination during nonclinical drug development," conducted at the 58th Annual Meeting of the Society of Toxicology, March 2019 in Baltimore, MD. In this paper, we further discuss and incorporate the perspectives of authors representing different roles, such as Study Director, Study Pathologist, Pharmacology/Toxicology Reviewer (U.S. Food and Drug Administration), and Sponsor in the determination and use of adversity. We also present a practical stepwise approach as an aid in this assessment, and further apply these principles to discuss 10 case studies with different therapeutic modalities and unique challenges.
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Desenvolvimento de Medicamentos , Avaliação Pré-Clínica de Medicamentos/métodos , Nível de Efeito Adverso não Observado , Preparações Farmacêuticas , Medição de Risco/métodos , Estados Unidos , United States Food and Drug AdministrationRESUMO
The INHAND (International Harmonization of Nomenclature and Diagnostic Criteria for Lesions) Project (www.toxpath.org/inhand.asp) is a joint initiative of the societies of toxicologic Pathology from Europe (ESTP), Great Britain (BSTP), Japan (JSTP), and North America (STP) to develop an internationally accepted nomenclature for proliferative and nonproliferative lesions in laboratory animals. The purpose of this publication is to provide a standardized nomenclature for classifying lesions observed in most tissues and organs from the dog used in nonclinical safety studies. Some of the lesions are illustrated by color photomicrographs. The standardized nomenclature presented in this document is also available electronically on the internet (http://www.goreni.org/). Sources of material included histopathology databases from government, academia, and industrial laboratories throughout the world. Content includes spontaneous lesions, lesions induced by exposure to test materials, and relevant infectious and parasitic lesions. A widely accepted and utilized international harmonization of nomenclature for lesions in laboratory animals will provide a common language among regulatory and scientific research organizations in different countries and increase and enrich international exchanges of information among toxicologists and pathologists.
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Animais de Laboratório , Animais , Bases de Dados Factuais , Cães , Europa (Continente) , JapãoRESUMO
AIM AND OBJECTIVE: To study the profile, indications, related complications, and predictors of decannulation and mortality in patients who underwent tracheostomy in the pediatric intensive care unit (PICU). MATERIALS AND METHODS: Retrospective analysis of prospectively collected data of tracheostomies was done on patients admitted at PICU. Demographics, primary diagnosis, indication of tracheostomy, and durations of endotracheal intubation, mechanical ventilation, and tracheostomy cannulation were recorded. The indication was recorded in one of the four categories-upper airway obstruction (UAO), central neurological impairment (CNI), prolonged mechanical ventilation, and peripheral neuromuscular disorders). RESULTS: Two hundred ninety cases were analyzed. UAO (42%) and CNI (48.2%) were main indications in the halves of the study period, respectively. Decannulation was successful in 188 (64.8%) patients. Seventy-seven percentage UAO patients were decannulated successfully [OR (odds ratio); 95% CI (confidence interval), 2.647; 1.182-5.924, p = 0.018]. Age <1 year (0.378; 0.187-0.764; p = 0.007), nontraumatic, noninfectious central neurological diseases (0.398; 0.186-0.855; p = 0.018), and malignancy (0.078; 0.021-0.298; p <0.001), durations of posttracheostomy ventilation (0.937; 0.893-0.983; p = 0.008), and stay in the PICU (0.989; 0.979-0.999; p = 0.029) were predictors of unsuccessful decannulation. There were 91 (31.4%) deaths. Age <1 year (2.39 (1.13-5.05; p = 0.02), malignancy (17.55; 4.10-75.11; p <0.001), durations of posttracheostomy ventilation (1.06; 1.006-1.10; p = 0.028), and hospital stay (1.007; 1.0-1.013; p = 0.043) were independent predictors of mortality. Indication of UAO favored survivor (0.24; 0.09-0.57; p <0.001). CONCLUSION: The indications for tracheostomy in children had changed over the years. Infancy, primary diagnosis, length of posttracheostomy ventilation, and stay in the PICU and hospital were independent predictors of decannulation and mortality. WHAT THIS ADDS: Similar to developed countries, the age at the time of tracheostomy and indication are changing. Inability to decannulate and mortality were associated with the age of a child at the time of tracheostomy, indication, medical diagnosis, and duration of postprocedure mechanical ventilation and stay in the hospital. HOW TO CITE THIS ARTICLE: Sachdev A, Chaudhari ND, Singh BP, Sharma N, Gupta D, Gupta N, et al. Tracheostomy in Pediatric Intensive Care Unit-A Two Decades of Experience. Indian J Crit Care Med 2021;25(7):803-811.
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The tuneability of oxygen containing groups in graphene oxide (GO) that controls physicochemical properties is highly desirable for device applications. In this context, the thermally reduced graphene oxide (r-GO) powders and spin coated thin films with varying sp2/sp3 carbon network have been prepared using highly exfoliated GO (synthesized using modified Hummer's method with an innovative conjunction of lyophilisation). The additional step of lyophilisation results in the formation of highly exfoliated and monodispersed GO nanosheets as evidenced from FESEM, TEM, XRD, and Raman, FT-IR and UV-Vis spectroscopy. Spectroscopic analysis revealed the systematic evolution of r-GO with tuneable structural, optical and electrical properties as results of varying annealing temperatures (100-400 °C), due to restoration of sp2 conducting carbon network i.e., the formation of new -CâC- network and Stones-Wales defect. The tuneability of physical properties is further corroborated by change in the resistance values, as evidenced through the current-voltage characteristics in GO thin film based lateral device structures with Ag and Al top contacts. Controlling physicochemical properties at relatively low processing temperature warrants the utilization of GO and r-GO in various electronic and optoelectronic devices.
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The 2014 annual National Toxicology Program (NTP) Satellite Symposium, entitled "Pathology Potpourri" was held in Washington, D.C., in advance of the Society of Toxicologic Pathology's 33rd annual meeting. The goal of this annual NTP Symposium is to present current diagnostic pathology or nomenclature issues to the toxicologic pathology community. This article presents summaries of the speakers' presentations, including diagnostic or nomenclature issues that were presented, along with select images that were used for audience voting and discussion. Some lesions and topics covered during the symposium included a pulmonary mucinous adenocarcinoma in a male B6C3F1 mouse; plexiform vasculopathy in Wistar Han (Crl:WI[Han]) rats; staging of the estrous cycle in rats and mice; peri-islet fibrosis, hemorrhage, lobular atrophy and inflammation in male Sprague-Dawley (SD) rats; retinal dysplasia in Crl:WI[Han] rats and B6C3F1 mice; multicentric lymphoma with intravascular microemboli and tumor lysis syndrome, and 2 cases of myopathy and vascular anomaly in Tg.rasH2 mice; benign thymomas in Crl:WI[Han] rats; angiomatous lesions in the mesenteric lymph nodes of Crl:WI[Han] rats; an unusual foveal lesion in a cynomolgous monkey; and finally a series of nomenclatures challenges from the endocrine International Harmonization of Nomenclature and Diagnostic Criteria (INHAND) Organ Working Group (OWG).
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Introduction Non-metric dental traits (NMDTs) are a fundamental data source in forensic dentistry. Nevertheless, the insufficiency of data regarding the occurrence of these traits has instigated the present research endeavor aimed at ascertaining the prevalence, sexual dimorphism, and extent of inter-trait correlations within the Maharashtrian population of India. The secondary objective was to determine the correlations between NMDTs, dentoskeletal malocclusion, and ABO blood groups. Materials and methods This prospective, observational study included 528 individuals aged 18-30 years with dentoskeletal Class I, II, and III malocclusions. NMDTs such as the presence of Cusp of Carabelli (CoC) on the upper first molars, hypocone on the upper second molars, and tri- or bicuspid lower second premolars were observed on the dental casts of all individuals. The dental relationship was assessed clinically according to Angle's system for the classification of malocclusion. The skeletal relationship was assessed using lateral cephalograms of the individuals. ABO blood groups were obtained from their medical records. The Chi-square test of independence was used to assess the associations between various variables. The correlation between each measurement was determined using Spearman's correlation test. Multivariate analysis enabled the identification of parameters that exhibited independent associations with NMDTs. A multinomial logistic regression model was constructed using NMDTs as the outcome variable. Results The mean age of males was 20.82 ± 1.71 years and 21.15 ± 1.76 years was in females. NMDTs were predominantly seen in females (n=394, 75%), with Class II dentoskeletal malocclusion (n=265, 50%) and B blood group ((n=199, 38%). All traits showed bilateral predominance. A statistically significant association was found between CoC, dentoskeletal malocclusion, hypocone, and tricuspid lower second premolars (p <0.05). All NMDTs showed a negative correlation with sex, a positive correlation between age and the presence of hypocones and CoC, a negative correlation between age and tricuspid lower second premolars, a strong positive correlation with dentoskeletal malocclusion, and a weak positive correlation with ABO blood groups. Multinomial logistic regression model analysis revealed that none of the independent variables were statistically significant predictors of the presence of CoC and tricuspid lower second premolars, while dentoskeletal malocclusion and sex were significant predictors of the presence of the hypocone trait. Conclusion NMDTs showed a female predilection with bilateral predominance. A significant association was observed between these traits and dentoskeletal malocclusions. The most commonly observed NMDT was the presence of a hypocone on the upper second molars, followed by the tricuspid lower second premolars and the CoC.
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Many proteins that self-assemble into amyloid and amyloid-like fibers can adopt diverse polymorphic forms. These forms have been observed both in vitro and in vivo and can arise through variations in the steric-zipper interactions between ß-sheets, variations in the arrangements between protofilaments, and differences in the number of protofilaments that make up a given fiber class. Different polymorphs arising from the same precursor molecule not only exhibit different levels of toxicity, but importantly can contribute to different disease conditions. However, the factors which contribute to formation of polymorphic forms of amyloid fibrils are not known. In this work, we show that in the presence of 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine, a highly abundant lipid in the plasma membrane of neurons, the aggregation of α-synuclein is markedly accelerated and yields a diversity of polymorphic forms under identical experimental conditions. This morphological diversity includes thin and curly fibrils, helical ribbons, twisted ribbons, nanotubes, and flat sheets. Furthermore, the amyloid fibrils formed incorporate lipids into their structures, which corroborates the previous report of the presence of α-synuclein fibrils with high lipid content in Lewy bodies. Thus, the present study demonstrates that an interface, such as that provided by a lipid membrane, can not only modulate the kinetics of α-synuclein amyloid aggregation but also plays an important role in the formation of morphological variants by incorporating lipid molecules in the process of amyloid fibril formation.
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Amiloide , alfa-Sinucleína , alfa-Sinucleína/química , Amiloide/química , Membrana Celular/metabolismo , Corpos de Lewy/metabolismo , LipídeosRESUMO
Introduction Obstructive Airway Diseases (OADs) are the leading cause of death among chronic respiratory diseases worldwide, and novel therapies are direly needed. Fluticasone furoate/vilanterol (FF/Vi) (100/25 µg) is the first once-daily ICS/uLABA marketed in India for COPD since 2021. Considering its limited real-world experience in OAD patients in Indian clinical settings, a large drug utilization study (DUS) was planned. Methodology We conducted a cross-sectional, observational DUS at 1900 outpatient clinics in India from October 2021 to March 2022. Prescription data and medical history of patients who were prescribed the FF/Vi combination were collected. Results It was observed that FF/Vi was prescribed in an almost equal number of patients with COPD (44.2%) and asthma (42.9%). The majority of the patients (74%) were switched from previous ICS/LABA to this ICS/uLABA, while 26% of patients were treatment naïve. The average CAT score was 19.5±7.8 (43.2% GOLD Group C and 32.2% GOLD Group B) in COPD patients, while the average ACQ-5 score was 2.6±1.3 (33.1% GINA Step 3, 29.5% GINA Step 2) in asthmatic patients. Most of the patients (63.9%) had raised biomarkers (Blood eosinophil count >300 cells/µl). Prior history of exacerbation was present in 65% of patients with annual exacerbation rates of 1.2 in COPD, 1.1 in asthma, and 1.2 in asthma-COPD overlap syndrome (ACOS). Leukotriene inhibitors (42%) and LAMAs (30.8%) were common add-on medications. Conclusion We observed a trend towards a shift to once-daily ICS/uLABA (FF/Vi) by physicians, especially in symptomatic and exacerbating OAD patients with underlying comorbidities.
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Low density of formal care providers in rural India results in restricted and delayed access to standardized management of hypertension. Task-sharing with pharmacies, typically the first point of contact for rural populations, can bridge the gap in access to formal care and improve health outcomes. In this study, we implemented a hypertension care program involving task-sharing with twenty private pharmacies between November 2020 and April 2021 in two blocks of Bihar, India. Pharmacists conducted free hypertension screening, and a trained physician offered free consultations at the pharmacy. We calculated the number of subjects screened, initiated on treatment (enrolled) and the change in blood pressure using the data collected through the program application. Of the 3403 subjects screened at pharmacies, 1415 either reported having a history of hypertension or had elevated blood pressure during screening. Of these, 371 (26.22%) were enrolled in the program. Of these, 129 (34.8%) made at least one follow-up visit. For these subjects, the adjusted average difference in systolic and diastolic blood pressure between the screening and follow-up visits was -11.53 (-16.95 to -6.11, 95% CI) and -4.68 (-8.53 to -0.82, 95% CI) mmHg, respectively. The adjusted odds of blood pressure being under control in this group during follow-up visits compared to screening visit was 7.07 (1.29 to 12.85, 95% CI). Task-sharing with private pharmacies can lead to early detection and improved control of blood pressure in a resource-constrained setting. Additional strategies to increase patient screening and retention rates are needed to ensure sustained health benefits.
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Hipertensão , Farmácias , Humanos , Estudos Retrospectivos , População Rural , Hipertensão/diagnóstico , Hipertensão/tratamento farmacológico , Hipertensão/epidemiologia , Pressão Sanguínea/fisiologiaRESUMO
The risk of hepatocellular carcinoma increases with the persistence of non-alcoholic fatty liver disease. Triacylglycerol synthesis is initiated by glycerol-3-phosphate acyltransferase (GPAT). Of four isoforms, GPAT1 contributes 30-50% of total liver GPAT activity, and we hypothesized that it might influence liver susceptibility to tumorigenesis. C57Bl/6 mice deficient in GPAT1 were backcrossed 6 times to C3H mice. After exposure to the carcinogen diethylnitrosamine (DEN) and the tumor promoter phenobarbital, male Gpat1â»/â» mice, compared with controls (Gpat1âº/âº), had 93% fewer macroscopically visible nodules per liver at 21 weeks of age and 39% fewer at 34 weeks of age. Microscopically, control mice had increased numbers of foci of altered hepatocytes, particularly the basophilic subtype, as well as more, and malignant, liver neoplasms than did the Gpat1â»/â» mice. At 21 weeks of age, 50% (4/8) of control mice (50%) had hepatocellular adenomas with an average multiplicity (tumors per tumor-bearing-animal) of 4.3, while none occurred in 8 Gpat1â»/â» mice. At 34 weeks of age, all 15 control mice (100%) had hepatocellular adenomas with an average multiplicity of 5.2 compared to an incidence of 93% in Gpat1â»/â» mice and multiplicity of 3.1. HCCs were observed in 13% of control mice and in only 6% of Gpat1â»/â» mice. These data show that alterations in the formation of complex lipids catalyzed by Gpat1 reduce susceptibility to DEN-induced liver tumorigenesis.
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Glicerol-3-Fosfato O-Aciltransferase/deficiência , Neoplasias Hepáticas Experimentais/enzimologia , Animais , Proliferação de Células , Dietilnitrosamina/toxicidade , Predisposição Genética para Doença , Glicerol-3-Fosfato O-Aciltransferase/genética , Glicerol-3-Fosfato O-Aciltransferase/metabolismo , Hepatócitos/citologia , Hepatócitos/metabolismo , Hepatócitos/patologia , Histocitoquímica , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas Experimentais/induzido quimicamente , Neoplasias Hepáticas Experimentais/genética , Neoplasias Hepáticas Experimentais/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neoplasias Experimentais , Tamanho do Órgão , PPAR alfa/metabolismo , Fenobarbital/toxicidade , RNA Mensageiro , Estatísticas não ParamétricasRESUMO
OBJECTIVES: The objective of study was to find an association between the timing of tracheostomy with duration of mechanical ventilation (MV) and length of stay (LOS) in pediatric intensive care unit (PICU) and hospital. METHODS: The data were collected prospectively from 2000 to 2018 and were analyzed retrospectively. Data included clinical diagnosis, indication, and duration (days) of MV, LOS in PICU and hospital before and after tracheostomy. Patients who did not receive MV or underwent MV for <24 h were excluded. According to the indication of tracheostomy enrolled patients were divided into four groups-airways anomalies (AA), central neurological impairment (CNI), cardiopulmonary insufficiency (CPI), and neuromuscular disorders (NMD). Patients in each group were divided into early (ET) and late tracheostomy (LT) category based on the median (interquartile range interquartile range [IQR]) days of pretracheostomy MV. RESULTS: Two hundred and fifty six patients were analyzed. The frequency and median [IQR] days of pretracheostomy MV were -AA 54 [7(3,16)], CNI 120 [12(9,16)], CPI 51 [25(16.5,30.5)], and NMD 31[12(8,16.5)]. In AA patients, median (IQR) durations of posttracheostomy MV [2(1,5.2) versus 3.5(2,12); p = 0.032], PICU [7(5,8.2) versus11(7,18); p = 0.004] and hospital [12(9.7,21) versus 21.5(12,28); p = 0.027] stays were lower in ET as compared with LT group. Posttracheostomy MV duration was significantly short in ET patients with CNI and NMD (p < 0.005). The total days of MV, PICU and hospital stay were significantly lower in ET as compared with LT patients in all four groups (p < 0.01). CONCLUSION: As compared with LT, ET patient had shorter durations of total MV and PICU and hospital stay.
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Unidades de Terapia Intensiva Pediátrica , Traqueostomia , Criança , Humanos , Unidades de Terapia Intensiva , Tempo de Internação , Respiração Artificial , Estudos RetrospectivosRESUMO
Aggregation of alpha-synuclein (α-Syn) drives Parkinson's disease (PD), although the initial stages of self-assembly and structural conversion have not been directly observed inside neurons. In this study, we tracked the intracellular conformational states of α-Syn using a single-molecule Förster resonance energy transfer (smFRET) biosensor, and we show here that α-Syn converts from a monomeric state into two distinct oligomeric states in neurons in a concentration-dependent and sequence-specific manner. Three-dimensional FRET-correlative light and electron microscopy (FRET-CLEM) revealed that intracellular seeding events occur preferentially on membrane surfaces, especially at mitochondrial membranes. The mitochondrial lipid cardiolipin triggers rapid oligomerization of A53T α-Syn, and cardiolipin is sequestered within aggregating lipid-protein complexes. Mitochondrial aggregates impair complex I activity and increase mitochondrial reactive oxygen species (ROS) generation, which accelerates the oligomerization of A53T α-Syn and causes permeabilization of mitochondrial membranes and cell death. These processes were also observed in induced pluripotent stem cell (iPSC)-derived neurons harboring A53T mutations from patients with PD. Our study highlights a mechanism of de novo α-Syn oligomerization at mitochondrial membranes and subsequent neuronal toxicity.
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Doença de Parkinson , alfa-Sinucleína , Cardiolipinas/metabolismo , Humanos , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Neurônios/metabolismo , Neurônios/patologia , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , alfa-Sinucleína/metabolismoRESUMO
Toxicogenomics is the application of toxicology, genetics, molecular biology and environmental health to describe the response of organisms to environmental stimuli. The field of toxicogenomics has developed over the past 15 years mainly due to advances in toxicology, molecular genetics and cell biology. Its prospective use to resolve crucial data gaps and data inconsistencies could improve risk assessment by providing additional data to increase the understanding of mechanisms and modes of action (MOA) and enhance the reliability of dose-response extrapolation. Thus, toxicogenomics holds promise for advancing the scientific basis of risk assessments. However, one of the current issues is how genomic/transcriptional data is being used to further describe a MOA for oncogenicity and, in turn, its potential uses in cancer risk assessment. This commentary identifies how toxicogenomics could be used on a case by case basis to add information to a MOA addressing both the opportunities and challenges this technology holds. In addition, some pitfalls to avoid in the generation and interpretation of toxicogenomic data and validation issues that need to be addressed before toxicogenomics can be used in the risk assessment process and regulatory decisions are discussed.
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Genômica/métodos , Toxicogenética/métodos , Toxicologia/métodos , Animais , Transformação Celular Neoplásica/genética , Relação Dose-Resposta a Droga , Genômica/tendências , Humanos , Reprodutibilidade dos Testes , Medição de Risco/métodos , Medição de Risco/tendências , Toxicogenética/tendências , Toxicologia/tendênciasRESUMO
We report the fabrication of single-phase polycrystalline Pb0.85Bi0.10(Zr0.52Ti0.48)O3 (PBiZT) ceramic which shows large polarization, i.e., â¼40 µC cm-2 and piezoelectric coefficients â¼130 pC N-1 and giant linear change in capacitive reactance and dielectric properties with increasing and decreasing pressure in the range of 1 kHz to 5 MHz. Nearly 70% change in dielectric constant and 56% change in capacitive reactance were obtained in the pressure range of 20-200 MPa, which makes it suitable for applications as a capacitive pressure sensor/gauge. The sensitivity of the device is calculated as 0.66 MPa-1 and 18.2 MPa-1 at 1 MHz and 5 MHz, respectively, which is the highest ever reported value so far for any bulk polycrystalline ceramic. The compressive stress of the device was tested according to the standard test method as a function of linear and volumetric strain, which yields the Young's modulus, Bulk modulus, and Poisson's ratio of the device. These values were further utilized to calculate actual stress in the sample and energy density using ANSYS software, which indicates at least four orders smaller pressure in the sample compared to the applied pressure.
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The fibronectin type II (FnII) domain, present in diverse vertebrate proteins, plays crucial roles in several fundamental biological processes. PDC-109, the major bovine seminal plasma protein, contains two FnII domains that bind to choline phospholipids on sperm plasma membrane and induce lipid efflux crucial for successful fertilization. PDC-109 also exhibits chaperone-like activity and protects other proteins against various types of stress. Here, we show that a core tryptophan residue is highly conserved across species in the FnII domains. Mutation of conserved tryptophan residues W47, W93, and W106 in the FnII domains of PDC-109 to alanine leads to drastic decrease or complete abolition of membrane-binding and chaperone-like activities. These observations suggest that conserved tryptophans are important for the function of FnII proteins.
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Membrana Celular/metabolismo , Sequência Conservada , Proteínas Secretadas pela Vesícula Seminal/química , Proteínas Secretadas pela Vesícula Seminal/metabolismo , Triptofano , Sequência de Aminoácidos , Animais , Bovinos , Ligantes , Modelos Moleculares , Mutação , Domínios Proteicos , Multimerização Proteica , Estrutura Quaternária de Proteína , Proteínas Secretadas pela Vesícula Seminal/genéticaRESUMO
RATIONALE: Epitopes were delineated for allergenic proteins, but studies are required to identify residues mediating IgE binding. In the present study, the in silico approach was used to identify IgE-binding residues of Alt a 13(1-50) fragment and confirmed by experimental approach. METHOD AND RESULTS: IgE-binding epitopes of Alt a 13 mapped computationally were cloned, expressed, purified, and characterized using various immunochemical and biophysical methods. Among four fragments of Alt a 13, Alt a 13(1-50) demonstrated maximum IgE binding with two immunodominant regions and was mutated at these regions. The mutation in first region, Alt a 13(1-50)-K4A_S6F, did not show any change in immunological and biophysical properties of protein. However, mutations in the second region, Alt a 13(1-50)-T21F_N27I, caused reduced IgE binding, histamine release, and low IL-4 release on stimulation of Alternaria alternata positive patients peripheral blood mononuclear cells in vitro. CONCLUSION: This suggests that residues T21 and N27 are important for the secondary structure. In conclusion, Alt a 13(1-50)-T21F_N27I with reduced Th 2 response and intact T-cell proliferation capacity has potential for clinical use.
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Alternaria/imunologia , Antígenos de Fungos/imunologia , Epitopos/imunologia , Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Linfócitos T/imunologia , Adolescente , Adulto , Anticorpos Antifúngicos/sangue , Antígenos de Fungos/química , Antígenos de Fungos/genética , Biologia Computacional , Mapeamento de Epitopos , Epitopos/química , Epitopos/genética , Feminino , Liberação de Histamina/imunologia , Humanos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Proteínas Mutantes/genética , Proteínas Mutantes/imunologia , Proteínas Mutantes/metabolismo , Mutação/genética , Fragmentos de Peptídeos/imunologia , Fragmentos de Peptídeos/metabolismo , Engenharia de Proteínas , Estrutura Secundária de Proteína , Análise de Sequência de Proteína , Testes Cutâneos , Adulto JovemRESUMO
BACKGROUND: Immunotherapy (IT) is practiced mainly with mixed and single allergen vaccines. But studies are rare with mixed allergen preparations. OBJECTIVE: The objective of this study is to study mix and single insect allergen IT in patients of allergic rhinitis and asthma. METHODS: We performed a double-blind placebo-controlled trial of mix and single allergen IT for 1 year in 99 patients of asthma or rhinitis or both. There were two groups, (1) active allergen IT (n = 61) with three subgroups single insect extract (cockroach, housefly, or mosquito) and mix allergen IT (two or three insect extracts) and (2) placebo (n = 38). Clinical (skin reactivity, airway reactivity, and symptom score) and immunological (IgE/IgG4 and IgG1/IgG4 ratio) parameters were assessed at baseline and after 1 year of IT. RESULTS: Eighty-five patients completed 1 year of IT. The active allergen IT group patients showed a significant improvement compared to baseline values (p < 0.05) and placebo group patients (p < 0.05) with regard to symptom scores, FEV1 values, and immunological parameters (IgG4). No significant difference was found between mixed and single IT group patients for changes in clinical and immunological parameters. Positive correlation was observed between increase in IgG4 and clinical improvement. The changes in above parameters in placebo group were nonsignificant after 1 year of treatment. CONCLUSION: IT with two to three mix extract from the same allergen group is effective for insect hypersensitivity.
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Alérgenos/imunologia , Asma/imunologia , Dessensibilização Imunológica , Proteínas de Insetos/imunologia , Rinite Alérgica Perene/imunologia , Adolescente , Adulto , Animais , Asma/sangue , Asma/fisiopatologia , Asma/terapia , Extratos Celulares/administração & dosagem , Extratos Celulares/efeitos adversos , Baratas/imunologia , Misturas Complexas/administração & dosagem , Misturas Complexas/efeitos adversos , Culicidae/imunologia , Progressão da Doença , Feminino , Moscas Domésticas/imunologia , Humanos , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Testes de Função Respiratória , Rinite Alérgica Perene/sangue , Rinite Alérgica Perene/fisiopatologia , Rinite Alérgica Perene/terapiaRESUMO
Genetic engineering of food crops has significantly influenced the agricultural productivity over the past two decades. It has proved a valuable tool, offering crops with higher yields, improved nutritional quality, resistance against pesticides, herbicides and tolerance against abiotic stresses. However, the safety assessment of genetically engineered (GE) crops is prerequisite before introduction into human food chain. The present study was aimed to assess the toxicity and allergenicity of leaf curl virus resistant GE tomato compared to its wild-type species. Balb/c mice fed with genetically engineered or wild-type tomato did not show significant differences in growth, body weight (P > 0.05) and food consumption when compared with control mice. Values for serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase, urea and cholesterol were comparable in GE and wild-type tomato fed mice. Mice immunized with GE or wild-type tomato extract showed low IgE response. Lung histology of ovalbumin fed mice showed bronchoconstriction with eosinophilic infiltration whereas GE or wild-type tomato showed no cellular infiltration with normal airways. Genetically engineered and wild-type tomato sensitized mice demonstrated similar IL-4 release in splenic cell culture supernatant. GE and wild tomato extract on ELISA showed comparable IgE binding (P > 0.05) with food allergic patients' sera. In conclusion, genetically engineered tomato showed no toxicity in mice and allergenicity is similar to the wild-type tomato.
Assuntos
Plantas Geneticamente Modificadas/efeitos adversos , Plantas Geneticamente Modificadas/genética , Solanum lycopersicum/efeitos adversos , Solanum lycopersicum/genética , Alérgenos/imunologia , Animais , Begomovirus/genética , Hipersensibilidade Alimentar/sangue , Humanos , Imunoglobulina E/imunologia , Solanum lycopersicum/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Soro/imunologia , Baço/imunologiaRESUMO
BACKGROUND: Previous studies elsewhere have shown higher serum immunoglobulin E (IgE) levels in smokers and an association between smoking and sensitisation to allergens. Such information is not available for Indian population. Hence, the present study was carried out to evaluate the effect of smoking on atopic predisposition and sensitisation to aeroallergens. METHODS: A total of 70 subjects were included in the present study comprising of 25 smokers, 22 reformed smokers and 23 non-smokers. Absolute eosinophil count (AEC), serum total IgE levels and skin prick test (SPT) against common aeroallergens were performed in all the subjects along with breath carbon monoxide (CO) levels and pulmonary function tests. RESULTS: Smokers showed significantly higher serum total IgE levels (328.80 +/- 161.82 IU/mL) as compared to reformed smokers 177.27 +/- 86.47 IU/mL) and non-smokers (29.56 +/- 9.75 IU/mL). A number of subjects among smokers and reformed smokers elicited positive SPT reactions to various allergen extracts. Non-smokers did not show any significant positive skin reaction. The AECs were slightly higher in smokers (350 +/- 1145.61/mm3) as compared to reformed smokers (305 +/- 146.33/ mm3). Breath CO was considerably higher (greater than three times) in smokers than reformed smokers. However, reformed smokers showed greater airways obstruction than smokers. The former also had higher Brinkman index (646.81 +/- 392.32) as compared to the latter (448.36 +/- 279.86). CONCLUSIONS: Smokers had significantly higher IgE serum levels than reformed smokers and non-smokers. Smoking seems to induce an atopic orientation and allergen sensitisation in individuals.