RESUMO
CONTEXT: Atrial fibrillation (AF) is the most common sustained arrhythmia in clinical practice. The increasing evidence supports links between inflammation and AF. There is evidence showing that obesity is a major cause of adipose tissue (AT) inflammation. Ghrelin (GHRL), through its growth hormone secretagogue receptor (GHS-R) present on adipose tissue macrophages (ATMs), could modulate AT inflammation. OBJECTIVE: Our study aimed to evaluate the role of adipose tissue macrophages (ATMs) and their GHS-R in adipose tissue samples of right atrial appendages (RAA) biopsies. SUBJECTS AND METHOD: We obtained RAA biopsies from 10 obese patients, undergoing cardiac surgery for coronary artery bypass graft (CABG) and developing postoperative atrial fibrillation (POAF). The epicardial tissue samples were examined using immunohistochemistry to visualize and quantify CD68 and GSH-R expression of the ATMs. RESULTS: Histologically, the mean adipocyte diameter (MAD) of epicardial adipose tissue (EAT) was larger in EAT samples with inflammation as compared to EAT without inflammation (84.2 µm vs. 79.6 µm). The expression of CD68 was lower in EAT without inflammation as compared to EAT with inflammation in adipose tissue samples. Similarly, the expression of GSH-R was lower in EAT samples without inflammation as compared to EAT samples with inflammation in adipose tissue. CONCLUSIONS: Increased epicardial fat area, macrophage infiltration, and GHS-R expression in epicardial ATMs appeared to be associated with postoperative atrial fibrillation in obese patients.
Assuntos
Infecções por Vírus Epstein-Barr/diagnóstico , Herpesvirus Humano 4/fisiologia , Síndromes de Imunodeficiência/diagnóstico , Células Matadoras Naturais/imunologia , Receptores de IgG/genética , Citotoxicidade Celular Dependente de Anticorpos , Células Cultivadas , Criança , Feminino , Humanos , Masculino , Linhagem , RNA Viral/análiseRESUMO
A variety of strategies have been designed for sequence-based HLA typing (SBT) and for the isolation of new human leucocyte antigen (HLA) alleles, but unambiguous characterization of complete genomic sequences remains a challenge. We recently reported a simple method for the group-specific amplification (GSA) and sequencing of a full-length C*04 genomic sequence in isolation from the accompanying allele. Here we build on this strategy and present homologous methods that enable the isolation of HLA-C alleles belonging to another two allele groups. Using this approach, which can be applied to sequence-based typing in some clinical settings, we have successfully characterized three novel HLA-C alleles (C*04:128, C*07:01:01:02, and C*08:62).
Assuntos
Alelos , Antígenos HLA-C/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico , Regiões 5' não Traduzidas , Sequência de Bases , Éxons , Antígenos HLA-C/genética , Antígenos HLA-C/imunologia , Teste de Histocompatibilidade , Humanos , Íntrons , Modelos Moleculares , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
To determine the complete sequence of a newly identified human leukocyte antigen (HLA)-C allele, we designed a method where the full genomic sequence of HLA-C*04 was amplified in isolation from the patient second HLA-C allele in a single polymerase chain reaction (PCR), using primers spanning its 5'- and 3'-untranslated regions. The new allele, officially designated HLA-C*04:71, differs from HLA-C*04:01:01:01 by two single-nucleotide polymorphisms: one determines substitution of phenylalanine for serine 9 at the B pocket of the peptide-binding site; the second substitution is a new polymorphism in intron 5. Phe-9 is characteristic of Cw1 alleles and its presence in C*04:71 most likely affects its peptide-binding repertoire. The principle we have used for C*04:71 isolation could be adapted for unambiguous sequence-based HLA-C typing of selected samples in a clinical setting.
Assuntos
Alelos , Antígenos HLA-C/genética , Peptídeos/química , Motivos de Aminoácidos , Sequência de Bases , Cristalografia por Raios X , Feminino , Genômica , Antígenos HLA-C/química , Humanos , Dados de Sequência Molecular , Ligação Proteica , Alinhamento de SequênciaRESUMO
Natural killer (NK) and T-lymphocytes monitor human leukocyte antigen (HLA)-E expression through CD94:NKG2 heterodimers. Structural polymorphism is not a hallmark for NK-complex genes on chromosome 12, except for complete NKG2C deletion in some humans. We present a method for fast, simple and accurate assessment of NKG2C copy-number variation - presence or absence in the genome of an NKG2C gene, in homo- or heterozygosis, is detected by a single conventional polymerase chain reaction that yields amplicons of different lengths in each genotype. We have also determined the NKG2C genotypes of a reference cell panel comprising 13 NK- and tumour-cell lines and 39 Epstein-Barr virus transformed cells from the International Histocompatibility Workshop. Our results should facilitate research on the importance of NKG2C and its deletion for immunity.
Assuntos
Variações do Número de Cópias de DNA/genética , Células Matadoras Naturais/metabolismo , Subfamília C de Receptores Semelhantes a Lectina de Células NK/genética , Reação em Cadeia da Polimerase/métodos , Linfócitos T/metabolismo , Linhagem Celular Transformada , Linhagem Celular Tumoral , Cromossomos Humanos Par 12/genética , Primers do DNA , Dosagem de Genes , Herpesvirus Humano 4/genética , Heterozigoto , Homozigoto , Humanos , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Subfamília C de Receptores Semelhantes a Lectina de Células NK/imunologia , Linfócitos T/citologia , Linfócitos T/imunologiaRESUMO
The genetic susceptibility to multiple sclerosis (MS) is only partially explained, and it shows geographic variations. We analyse here two series of Spanish patients and healthy controls and show that relapsing MS (R-MS) is associated with a gene deletion affecting the hypothetically soluble leukocyte immunoglobulin (Ig)-like receptor A3 (LILRA3, 19q13.4), in agreement with an earlier finding in German patients. Our study points to a gene-dose-dependent, protective role for LILRA3, the deletion of which synergizes with HLA-DRB1(*)1501 to increase the risk of R-MS. We also investigated whether the risk of suffering R-MS might be influenced by the genotypic diversity of killer-cell Ig-like receptors (KIRs), located only approximately 400 kb telomeric to LILRA3, and implicated in autoimmunity and defence against viruses. The relationship of LILRA3 deletion with R-MS is not secondary to linkage disequilibrium with a KIR gene, but we cannot exclude some contributions of KIR to the genetic susceptibility to R-MS.
Assuntos
Deleção de Genes , Antígenos HLA-DR/genética , Esclerose Múltipla/genética , Receptores Imunológicos/genética , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Criança , Feminino , Variação Genética , Genótipo , Cadeias HLA-DRB1 , Humanos , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/epidemiologia , Receptores KIR/genética , Espanha/epidemiologia , Adulto JovemRESUMO
OBJECTIVES: Biclonal gammopathies are characterized by the clonal proliferation of plasma cells or their B-lymphoid progenitors and are associated with the production of abnormal immunoglobulins (M proteins or paraproteins). There are no known studies that have analyzed this disease in Spain. We studied the underlying diseases, characteristics of paraproteins and the evolution of a series of patients with biclonal gammopathy. MATERIAL AND METHODS: We reviewed clonal gammopathies at the Department of Immunology of Hospital Puerta de Hierro in Madrid, between 1970 and 2011, selecting those patients with biclonal gammopathy in one reading. We collected data on the patient's epidemiology, underlying disease, associated diseases, therapies and paraprotein and immunoglobulin levels. RESULTS: Of the 1626 cases of clonal gammapathies, 47 were biclonal gammopathy (2.89%). The median follow-up was 2 years. The main associated condition was biclonal gammopathies of undetermined significance (BGUS). The most common paraprotein combination was IgG-IgG. Upon conducting a second paraprotein reading, 81% of the patients had lost at least 1 monoclonal component. A third of the patients had not undergone treatment. CONCLUSIONS: Biclonal gammopathy are fundamentally associated with biclonal gammopathies of undetermined significance. No biclonal gammopathies of undetermined significance evolved to a malignant disease. In a high percentage of patients, at least 1 of the 2 clonal components disappeared, sometimes spontaneously.
RESUMO
Research on multiple sclerosis (MS) frequently requires typing for allele HLA-DRB1*1501, which the complexities of the HLA system can restrict to specialised histocompatibility laboratories. To overcome this limitation, we have implemented a simple, robust and highly specific method for DRB1*1501 detection. One single-tube polymerase-chain reaction (PCR) per DNA sample allows for detecting DR2 individuals. The spare PCR products of these are then sequenced to identify allele DRB1*1501 by comparison with the official, publicly accessible HLA database. This approach, much simpler than previously available methods, should facilitate research on MS by making accurate identification of DRB1*1501 accessible to neuroscience laboratories.
Assuntos
Testes Genéticos/métodos , Antígenos HLA-DR/genética , Teste de Histocompatibilidade/métodos , Esclerose Múltipla/diagnóstico , Esclerose Múltipla/genética , Alelos , Bases de Dados Genéticas/estatística & dados numéricos , Predisposição Genética para Doença , Antígeno HLA-DR2/genética , Cadeias HLA-DRB1 , Humanos , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Análise de Sequência de DNA/métodosRESUMO
UNLABELLED: Revolutionary progresses in computer technology are of great use during forensic investigations in terms of facial identification based on reconstructions. AIM: The purpose of the current study is to evaluate the use and limits of facial reconstruction by considering soft tissues' thickness determination. MATERIAL AND METHOD: The authors investigated the limits of three methods used for soft tissues' thickness determination on a sample group consisting of 9 cadavers, 30 ultrasound investigations and 27 cranial X-rays. The measurements were performed at standard anthropological landmarks following methods previously described in literature. RESULTS: Measurement values for each soft tissue landmark overlapped between sexes, females having greater soft tissue thickness at two sites: infraorbital and supraorbital notches. US measurements demonstrate a much larger dependence of the soft tissue thickness according to different body posture and imply compressing soft tissues with the transducer. X-rays determinations revealed more accurate values than US, but are nocive to the subjects and can only be performed in standard radiographic positions. For cadavers, different postmortem stages and supine position generated inadequate results. CONCLUSION: In order to increase the degree of accuracy of craniofacial approximation it is necessary to obtain a validated data set specific for the Romanian population. This will allow a better determination of facial measurements opening new perspectives in understanding the relation between physical properties and facial soft tissue.
Assuntos
Cefalometria/métodos , Face/diagnóstico por imagem , Antropologia Forense/métodos , Imageamento Tridimensional , Dobras Cutâneas , Cadáver , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Radiografia , Caracteres Sexuais , UltrassonografiaRESUMO
Electronmicroscopic studies performed on atrial and ventricular tissue fragments from 9 human embryos, 8 to 15 weeks old, showed that in certain myocardiocytes, especially in the atrial ones, from the 10 weeks old embryos, there appear granules of secretion, very similar in morphology with those from adults, which had been demonstrated to contain the atrial natriuretic factor. The precocious appearance of this peptide represents a functional adaptation by which the embryo attempts at counterbalancing the actions of renin and angiotensin from the maternal circulation.
Assuntos
Glândulas Endócrinas/ultraestrutura , Coração/embriologia , Miocárdio/ultraestrutura , Fator Natriurético Atrial/metabolismo , Técnicas Citológicas , Glândulas Endócrinas/embriologia , Glândulas Endócrinas/metabolismo , Idade Gestacional , Histocitoquímica , Humanos , Microscopia Eletrônica , Miocárdio/metabolismoRESUMO
Electron microscopic investigations performed on heart tissue fragments from 12 human embryos. 8 to 15 weeks of age, showed that at the age of 8 weeks the myocardiocytes are poorly differentiated cells: they contain microfilaments of actin and myosin inserted on the cytoplasmic aspect of plasmalemma, a smooth endoplasmic reticulin and mitochondria, accounting for the early contraction. Gap junctions appear among myocardiocytes, confirming the myogenic theory of heart contractions. Within 10 to 15 weeks, actin and myosin myofilaments organize in sarcomeres. Eberth disks appear and numerous mitoses are to be seen.