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1.
BMC Nephrol ; 21(1): 97, 2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32169051

RESUMO

BACKGROUND: Growing evidence has shown that the gut-renal connection and gut microbiota dysbiosis play a critical role in immunoglobulin A nephropathy (IgAN). However, the fecal microbiome profile in Chinese patients with IgAN remains unknown. A cross-sectional study was designed for the first time to investigate the fecal microbiota compositions in patients with primary IgAN in China and to evaluate the relationship between the fecal microbiome and IgAN clinical presentation. METHODS: Fecal samples were collected from 17 IgAN patients and 18 age-, sex-, and body mass index-matched healthy controls, and bacterial DNA was extracted for 16S ribosomal RNA gene sequencing targeting the V3-V4 region. RESULTS: Fecal samples from the IgAN patients and healthy controls showed differences in gut microbiota community richness and compositions. Compared to the healthy controls, IgAN patients at the phylum level had an increased abundance of Fusobacteria, but a decreased abundance of Synergistetes. The significantly increased genera in the IgAN group were Escherichia-Shigella, Hungatella, and Eggerthella, all of which possess pathogenic potential. Furthermore, the genus Escherichia-Shigella was negatively associated with the estimated glomerular filtration rate (eGFR) but was positively associated with the urinary albumin-to-creatinine ratio (uACR). However, the genus rectale_group was present in the IgAN group with a low abundance and was negatively associated with the uACR. Functional analysis disclosed that infection-related pathways were enriched in the IgAN group. CONCLUSIONS: We demonstrate that gut microbiota dysbiosis occurs in patients with IgAN, and that changes in gut bacterial populations are closely related to IgAN clinical features, suggesting that certain specific gut microbiota may be a potential therapeutic target for IgAN.

2.
Immunobiology ; 224(3): 339-346, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30975435

RESUMO

A deficiency of complement factor H may lead to excessive consumption of C3 and an increase in C3b deposition, which are important pathological characteristics of lupus nephritis. Complement factor H-related proteins (CFHRs), comprising CFHR1 to CFHR5 (CFHR1-5), are members of the wider factor H/CFHR family. Their role in lupus nephritis remains unclear. In this study, we compared circulating levels of CFHR1-5 in 152 patients diagnosed with lupus nephritis and 20 unrelated healthy individuals to explore the relationship between the expression of CFHR1-5 and development of the disease. We found that plasma levels of CFHR3 and CFHR5 were higher in patients with lupus nephritis than in healthy individuals; also, CFHR3 and CFHR5 concentrations increased with increasing systemic lupus erythematosus disease activity index (SLEDAI) values (P < 0.05). Pearson's and Spearman's correlation test results confirmed that plasma CFHR3 and CFHR5 levels in lupus nephritis patients were positively correlated with proteinuria and levels of creatinine (Cr) and anti-dsDNA (correlation coefficients = 0.491-0.717, P < 0.05), while they were negatively correlated with plasma C3 levels and eGFR [correlation coefficients = -(0.706-0.788), P < 0.05]. Receiver operating characteristic (ROC) curve analysis results confirmed that plasma CFHR3 and CFHR5 levels were predictive of SLEDAI values and disease end points (area under the curve = 0.664-0.884, P < 0.05), with patients with both high CFHR3 and high CFHR5 exhibiting the shortest progression-free survival. Thus, both CFHR3 and CFHR5 are of prognostic value in lupus nephritis status.


Assuntos
Proteínas Sanguíneas/metabolismo , Proteínas do Sistema Complemento/metabolismo , Nefrite Lúpica/metabolismo , Adolescente , Adulto , Anticorpos Antinucleares/sangue , Apolipoproteínas/metabolismo , Circulação Sanguínea , Estudos de Casos e Controles , Criança , Complemento C3/metabolismo , Proteínas Inativadoras do Complemento C3b/metabolismo , Creatinina/sangue , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteinúria , Adulto Jovem
3.
Zhonghua Wei Chang Wai Ke Za Zhi ; 21(11): 1255-1260, 2018 Nov 25.
Artigo em Chinês | MEDLINE | ID: mdl-30506536

RESUMO

OBJECTIVE: To investigate the efficacy and safety of the bladder training in male patients before urinary catheter removal after mid-low rectal cancer surgery. METHODS: This was a prospective, open, randomized controlled study. INCLUSION CRITERIA: male patients; pathologically diagnosed as mid-low rectal adenocarcinoma; distance from tumor lower edge to anal margin ≤10 cm; standard radical surgery for rectal cancer, including intestinal resection and regional lymph node dissection. EXCLUSION CRITERIA: previous history of benign prostatic hyperplasia or history of prostate surgery; bladder dysfunction such as dysuria and urinary retention before surgery; local resection of rectal tumor or extended resection. According to the above criteria, 92 patients who underwent colorectal surgery at the Union Hospital of Fujian Medical University from June to December 2016 were prospectively included. The patients were randomly divided into bladder training group (n=43) and bladder non-training group (n=49) according to the random number table method. The study was approved by the Ethics Committee of the Union Hospital of Fujian Medical University (ethical approval number: 2016KY005) and registered with the China Clinical Trial Registration Center (ChiCTR) (registration No.ChiCTR-IOR-16007995). The implementation of patient's treatment measures, the data collection and analysis were based on the three-blind principle, using envelopes for distribution concealment. In the bladder training group, bladder training was routinely performed from the first day after operation to catheter removal, and in bladder non-training group the catheter was kept open till its removal. The catheter was removed in the early morning at the 5th day after surgery, and the spontaneous urine output was recorded and the residual urine volume of the bladder was measured after the first urination. The international prostate symptom score (IPSS) was applied to evaluate the patient's urinary function before and after surgery. RESULTS: The age of whole group was (58.6±10.9) years old, the body mass index was (22.4±2.7) kg/m 2, and the distance from tumor lower edge to anal margin was (6.5±1.9) cm. The baseline data, such as age, body mass index, distance from tumor lower edge to anal margin, preoperative IPSS score, preoperative bladder residual urine volume, neoadjuvant radiotherapy and chemotherapy, preventive ileostomy and surgical procedure were not significantly different between two groups (all P>0.05). There was no significant difference in IPSS scores evaluated at the second day (3.6±4.0 vs. 3.5±3.4, t=0.128, P=0.899) and one month (3.7±2.9 vs. 3.0±3.1, t=1.113, P=0.269) after catheter removal between the bladder training group and bladder non-training group. No significant difference in the postoperative residual urine volume of bladder (media 44 ml vs. 24 ml, Z=-1.466, P=0.143), the first spontaneous urination volume (median 200 ml vs. 150 ml, Z=-1.228, P=0.219) after catheter removal, and postoperative hospital stay [(8.2±4.5) days vs. (9.1±5.5) days, t=-0.805, P=0.423] was found. Urinary infection rate was 20.9%(9/43) in the training group, which was even higher than 8.2%(4/49) in the non-training group, but the difference was not significant(χ²=3.077, P=0.079). No patient needed re-catheterization in either group. CONCLUSIONS: The routine bladder training after mid-low rectal cancer surgery does not improve the urinary function, and can not reduce the residual urine volume of bladder after catheter removal. This routine clinical practice is not helpful for the bladder function recovery after rectal cancer surgery.


Assuntos
Neoplasias Retais , Bexiga Urinária , Retenção Urinária , Idoso , China , Humanos , Laparoscopia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recuperação de Função Fisiológica , Neoplasias Retais/cirurgia , Neoplasias Retais/terapia , Resultado do Tratamento , Bexiga Urinária/cirurgia , Retenção Urinária/terapia
4.
Vaccine ; 36(41): 6144-6151, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30194004

RESUMO

Influenza vaccines are the most effective intervention to prevent the substantial public health burden of seasonal and pandemic influenza. Hemagglutinin (HA), as the main antigen in inactivated influenza vaccines (IIVs), elicits functional neutralizing antibodies and largely determines IIV effectiveness. HA potency has been evaluated by single-radial immunodiffusion (SRID), the standard in vitro potency assay for IIVs, to predict vaccine immunogenicity with a correlation to protective efficacy. We previously reported that limited trypsin digestion (LTD) selectively degraded stressed HA, so that an otherwise conformationally insensitive biophysical quantification technique could specifically quantify trypsin-resistant, immunologically active HA. Here, we demonstrate that isotope dilution mass spectrometry (IDMS), a method capable of quantifying the absolute HA concentration without reference antigen use, can be further expanded by adding LTD followed with precipitation to selectively quantify the active HA. We test the LTD-IDMS assay on H7N9 vaccines stressed by low pH, raised temperature, or freeze/thaw cycles. This method, unlike SRID, has no requirement for strain-specific reference antigens or antibodies and can generate potency values that correlate with SRID. Thus, LTD-IDMS is a promising alternative in vitro potency assay for influenza vaccines to complement and potentially replace SRID in a pandemic when strain specific reagents may not be readily available.


Assuntos
Subtipo H7N9 do Vírus da Influenza A/imunologia , Subtipo H7N9 do Vírus da Influenza A/patogenicidade , Vacinas contra Influenza/uso terapêutico , Influenza Humana/imunologia , Influenza Humana/prevenção & controle , Hemaglutininas/metabolismo , Humanos , Marcação por Isótopo , Espectrometria de Massas
5.
Medicine (Baltimore) ; 97(34): e12038, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30142855

RESUMO

Colorectal cancer (CRC) ranked 3rd for cancer incidence and 4th for cancer death worldwide. Despite the increasing number of CRC studies, the etiology is not yet clear. In this study, we investigated the effects of the dinner-to-bed time, post-dinner walk and sleep duration on the risk for CRC.We conducted a matched case-control study based on hospital population. We involved 166 patients had a newly histologically confirmed CRC without previous treatment and 166 healthy healthy residents matched by age and gender at Fujian Medical Union Hospital. A self-designed questionnaire was used to information on demographic characteristics, dinner-to-bed time, post-dinner walk, sleep duration, and other behavioral factors. Conditional logistic regression was used to calculate the odds ratio (OR) and 95% confidence intervals (95% CIs) to assess the effect of dinner-to-bed time, post-dinner walking, and sleep duration as well as their joint effect on the risk of CRC at different genders.The adjusted odds ratio (AOR) of CRC for subjects with shorter dinner-to-bed time (2.0-2.9 h) were 2.527 (95% CIs = 1.127-5.337), relative to those with longer dinner-to-bed time (≥4 h), the difference was statistically significant (P < .05). Post-dinner walk was associated with a significantly decreased CRC risk (AOR = 0.339, 95% CIs = 0.203-0.865) compared with post-dinner non-walk. Compared with 6-9 h of sleep duration, the risk OR of CRC were 3.843 (95% CIs = 2.767-7.800, P < .05) and 2.12 (95% CIs = 0.754-5.959, P > .05) for long (≥9 h) and short (<6 h) sleep duration. The risk of CRC individuals with shorter dinner-to-bed time and post-dinner non-walk caused higher risk than those with longer dinner-to-bed time and post-dinner walk (AOR = 3.361, 95% CIs = 2.043-6.316). The risk of CRC was 2.231 (95% CIs = 1.089-3.762, P < .001), with a shorter dinner-to-bed time and ≥9 hours of sleep duration.We found that shorter dinner-to-bed time (<3 h), post-dinner walk, and long sleep duration (≥9 h) were seems to be related to CRC and may increase the risk of CRC.


Assuntos
Neoplasias Colorretais/etiologia , Refeições/fisiologia , Período Pós-Prandial/fisiologia , Sono/fisiologia , Caminhada/fisiologia , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fatores de Risco , Inquéritos e Questionários , Fatores de Tempo
6.
Vaccine ; 36(21): 3010-3017, 2018 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-29680201

RESUMO

Influenza vaccines are the most effective intervention to prevent the substantial public health burden of seasonal and pandemic influenza. The capability of hemagglutinin (HA), the main antigen in inactivated influenza vaccines (IIVs), to elicit functional neutralizing antibodies determines IIV effectiveness. When HA is subjected to environmental stress during manufacturing or while stored prior to administration, such as low pH and temperature excursions, the HA immunological activity can be affected. Single-radial immunodiffusion (SRID), the standard in vitro potency assay for IIVs, is believed to specifically detect immunologically active HA and has been applied to evaluate HA stability against stress. Here we report that transient low pH treatment and freeze/thaw cycles with HA in PBS abolish SRID-quantified in vitro potency for all HAs of multiple influenza strains. Raised temperature substantially decreases in vitro potency with more extensive HA structural changes. Chemical stress and mechanical stress moderately change SRID in vitro potency values in a strain-dependent manner. Trypsin digestion, which selectively degrades stressed HA, followed by RP-HPLC quantification as a candidate alternative in vitro potency assay yields results comparable to SRID. Mouse immunogenicity studies confirm that HA stressed by transient low pH treatment does not elicit functional antibodies in vivo, nor does it have a measureable SRID value. However, HA stressed by raised temperature elicits high titers of functional antibodies in vivo despite substantial loss of SRID in vitro potency. This discrepancy between SRID in vitro potency and vaccine immunogenicity suggests that SRID may not reliably indicate IIV potency under all conditions. Further efforts to develop alternate potency assays that can better predict in vivo immunogenicity should continue along with additional studies exploring HA conformation, SRID values and consequent immunogenicity.


Assuntos
Armazenamento de Medicamentos/métodos , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/imunologia , Tecnologia Farmacêutica/métodos , Potência de Vacina , Animais , Anticorpos Antivirais/sangue , Feminino , Congelamento , Concentração de Íons de Hidrogênio , Vacinas contra Influenza/isolamento & purificação , Vacinas contra Influenza/efeitos da radiação , Camundongos Endogâmicos BALB C , Temperatura Ambiente , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologia , Vacinas de Produtos Inativados/isolamento & purificação , Vacinas de Produtos Inativados/efeitos da radiação
7.
PLoS One ; 12(9): e0183676, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28934237

RESUMO

The Qiongdongnan Basin is a strongly overpressured basin with the maximum pressure coefficient (the ratio of the actual pore pressure versus hydrostatic pressure at the same depth) over 2.27. However, there exists a widespread low-overpressure interval between the strong overpressure intervals in the Yanan Sag of western basin. The mechanisms of the low-overpressure interval are not well understood. Three main approaches, pore pressure test data and well-log analysis, pressure prediction based on the relationship between the deviation of the velocity and the pressure coefficients, and numerical modeling, were employed to illustrate the distribution and evolution of the low-overpressure interval. And we analyzed and explained the phenomenon of the low-overpressure interval that is both underlain and overlain by high overpressure internal. The low-overpressure interval between the strong overpressure intervals can be identified and modelled by drilling data of P-wave sonic and the mud weight, and the numerical modeling using the PetroMod software. Results show that the low-overpressure interval is mainly composed of sandstone sediments. The porosities of sandstone in the low-overpressure interval primarily range from 15%-20%, and the permeabilities range from 10-100 md. Analysis of the geochemical parameters of C1, iC4/nC4, ΔR3, and numerical modeling shows that oil and gas migrated upward into the sandstone in the low-overpressure interval, and then migrated along the sandstone of low-overpressure interval into the Yacheng uplift. The low-overpressure both underlain and overlain by overpressure resulted from the fluids migrating along the sandstones in the low-overpressure interval into the Yacheng uplift since 1.9Ma. The mudstone in the strong overpressure interval is good cap overlain the sandstone of low-overpressure interval, therefore up-dip pinchouts or isolated sandstone in the low-overpressure interval locating the migration path of oil and gas are good plays for hydrocarbon exploration.


Assuntos
Hidrocarbonetos , Pressão Hidrostática , Oceanos e Mares , China , Modelos Estatísticos , Porosidade
8.
Vaccine ; 34(29): 3388-95, 2016 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-27154389

RESUMO

Influenza vaccines are the primary intervention to prevent the substantial health burden of seasonal and pandemic influenza. Subunit and split influenza vaccines are formulated, released for clinical use, and tested for stability based on their content of immunologically active (capable of eliciting functional antibodies) hemagglutinin (HA). Single-radial immunodiffusion (SRID), the standard in vitro potency assay in the field, is believed to specifically detect immunologically active HA. We confirmed that, with conformationally homogeneous HA preparations, SRID specifically detected native, pre-fusion HA, which elicited influenza neutralizing and hemagglutination inhibiting (HI) antibodies in mice, and it did not detect low-pH stressed, post-fusion HA, which was selectively removed from the SRID gel during a blotting step and was significantly less immunologically active. This selective detection was due to the SRID format, not a conformational specificity of the sheep antiserum used in the SRID, as the same antiserum detected non-stressed and low-pH stressed HA similarly when used in an ELISA format. However, when low-pH stressed HA was mixed with non-stressed HA, SRID detected both forms in mixed immunoprecipitin rings, leading to over-quantification of pre-fusion HA. We previously reported that trypsin digestion of antigen samples selectively degrade stressed HA, so that an otherwise conformationally insensitive biophysical quantification technique, reversed-phase high pressure liquid chromatography (RP-HPLC), can specifically quantify trypsin-resistant, immunologically active, pre-fusion HA. Here, we report that trypsin digestion can also improve the specificity of SRID so that it can quantify immunologically active, pre-fusion HA when it is mixed with less immunologically active, post-fusion HA.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/isolamento & purificação , Imunodifusão , Vacinas contra Influenza/imunologia , Tripsina/química , Animais , Anticorpos Antivirais/sangue , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Ensaio de Imunoadsorção Enzimática , Feminino , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Concentração de Íons de Hidrogênio , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização , Infecções por Orthomyxoviridae/prevenção & controle , Sensibilidade e Especificidade , Ovinos , Potência de Vacina
9.
Vaccine ; 33(41): 5342-5349, 2015 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-26348403

RESUMO

Influenza vaccines are the primary intervention for reducing the substantial health burden from pandemic and seasonal influenza. Hemagglutinin (HA) is the most important influenza vaccine antigen. Subunit and split influenza vaccines are formulated, released for clinical use, and tested for stability based on an in vitro potency assay, single-radial immunodiffusion (SRID), which selectively detects HA that is immunologically active (capable of eliciting neutralizing or hemagglutination inhibiting antibodies in an immunized subject). The time consuming generation of strain-specific sheep antisera and calibrated antigen standards for SRID can delay vaccine release. The limitation in generating SRID reagents was evident during the early days of the 2009 pandemic, prompting efforts to develop more practical, alternative, quantitative assays for immunologically active HA. Here we demonstrate that, under native conditions, trypsin selectively digests HA produced from egg or mammalian cell in monovalent vaccines that is altered by stress conditions such as reduced pH, elevated temperature, or deamidation, leaving native, pre-fusion HA, intact. Subsequent reverse-phase high pressure liquid chromatography (RP-HPLC) can separate trypsin-resistant HA from the digested HA. Integration of the resulting RP-HPLC peak yields HA quantities that match well the values obtained by SRID. Therefore, trypsin digestion, to pre-select immunologically active HA, followed by quantification by RP-HPLC is a promising alternative in vitro potency assay for influenza vaccines.


Assuntos
Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Imunodifusão/métodos , Vírus da Influenza A/imunologia , Vacinas contra Influenza/imunologia , Potência de Vacina , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Concentração de Íons de Hidrogênio , Vírus da Influenza A/classificação , Temperatura Ambiente
10.
J Virol ; 88(20): 11802-10, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25078705

RESUMO

Respiratory syncytial virus (RSV) is the leading infectious cause of severe respiratory disease in infants and a major cause of respiratory illness in the elderly. There remains an unmet vaccine need despite decades of research. Insufficient potency, homogeneity, and stability of previous RSV fusion protein (F) subunit vaccine candidates have hampered vaccine development. RSV F and related parainfluenza virus (PIV) F proteins are cleaved by furin during intracellular maturation, producing disulfide-linked F1 and F2 fragments. During cell entry, the cleaved Fs rearrange from prefusion trimers to postfusion trimers. Using RSV F constructs with mutated furin cleavage sites, we isolated an uncleaved RSV F ectodomain that is predominantly monomeric and requires specific cleavage between F1 and F2 for self-association and rearrangement into stable postfusion trimers. The uncleaved RSV F monomer is folded and homogenous and displays at least two key RSV-neutralizing epitopes shared between the prefusion and postfusion conformations. Unlike the cleaved trimer, the uncleaved monomer binds the prefusion-specific monoclonal antibody D25 and human neutralizing immunoglobulins that do not bind to postfusion F. These observations suggest that the uncleaved RSV F monomer has a prefusion-like conformation and is a potential prefusion subunit vaccine candidate. Importance: RSV is the leading infectious cause of severe respiratory disease in infants and a major cause of respiratory illness in the elderly. Development of an RSV vaccine was stymied when a clinical trial using a formalin-inactivated RSV virus made disease, following RSV infection, more severe. Recent studies have defined the structures that the RSV F envelope glycoprotein adopts before and after virus entry (prefusion and postfusion conformations, respectively). Key neutralization epitopes of prefusion and postfusion RSV F have been identified, and a number of current vaccine development efforts are focused on generating easily produced subunit antigens that retain these epitopes. Here we show that a simple modification in the F ectodomain results in a homogeneous protein that retains critical prefusion neutralizing epitopes. These results improve our understanding of RSV F protein folding and structure and can guide further vaccine design efforts.


Assuntos
Anticorpos Neutralizantes/imunologia , Antígenos Virais/imunologia , Epitopos/imunologia , Vírus Sinciciais Respiratórios/imunologia , Humanos , Proteólise
11.
J Biol Chem ; 283(24): 16641-52, 2008 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-18411262

RESUMO

Gangliosides are key players in neuronal inhibition, with antibody-mediated clustering of gangliosides blocking neurite outgrowth in cultures and axonal regeneration post injury. In this study we show that the ganglioside GT1b can form a complex with the Nogo-66 receptor NgR1. The interaction is shown by analytical ultracentrifugation sedimentation and is mediated by the sialic acid moiety on GT1b, with mutations in FRG motifs on NgR1 attenuating the interaction. One FRG motif was developed into a cyclic peptide (N-AcCLQKFRGSSC-NH(2)) antagonist of GT1b, reversing the GT1b antibody inhibition of cerebellar granule cell neurite outgrowth. Interestingly, the peptide also antagonizes neurite outgrowth inhibition mediated by soluble forms of the myelin-associated glycoprotein (MAG). Structure function analysis of the peptide point to the conserved FRG triplet being the minimal functional motif, and mutations within this motif inhibit NgR1 binding to both GT1b and MAG. Finally, using gene ablation, we show that the cerebellar neuron response to GT1b antibodies and soluble MAG is indeed dependent on NgR1 function. The results suggest that gangliosides inhibit neurite outgrowth by interacting with FRG motifs in the NgR1 and that this interaction can also facilitate the binding of MAG to the NgR1. Furthermore, the results point to a rational strategy for developing novel ganglioside antagonists.


Assuntos
Gangliosídeos/metabolismo , Proteínas da Mielina/metabolismo , Motivos de Aminoácidos , Animais , Sítios de Ligação , Células COS/metabolismo , Análise por Conglomerados , Gangliosídeos/química , Gangliosídeos/genética , Humanos , Camundongos , Camundongos Knockout , Mutação , Ácido N-Acetilneuramínico/química , Neuritos/metabolismo , Proteínas Nogo
12.
Brain Res Mol Brain Res ; 133(2): 187-97, 2005 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-15710235

RESUMO

Members of the MRG family of G-protein coupled receptors (GPCRs) are expressed predominately in small diameter sensory neurons of the dorsal root ganglia (DRG) suggesting a possible role in nociception. However, the large expansion of this gene family in rodents, combined with the lack of strict rodent orthologs for many of the human MRG genes, limits the usefulness of rodent models to evaluate human MRG involvement in nociception. Furthermore, the high degree of similarity between related rodent Mrg genes suggests that pharmacological approaches to define the function of individual receptors will prove difficult. The creation of an animal model to examine human MRG function will, therefore, require the identification of human MRG orthologs in a non-rodent species. Here we report the identification of MRGD, MRGE, and several MRGX orthologs in the crab-eating macaque, Macaca fascicularis. Similar to their human counterparts, all isolated macaque genes were expressed in dorsal root ganglia neurons. In the case of macaque MrgX2 and MrgD, expression was co-localized with the known nociceptive neuronal markers, IB4, VR1, and SP. Although expression in DRG neurons was the prominent feature of this family, we also found that MrgE was expressed in numerous brain regions of macaque, mouse, and human.


Assuntos
Expressão Gênica/fisiologia , Neurônios/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Animais , Northern Blotting/métodos , Southern Blotting/métodos , Encéfalo/anatomia & histologia , Encéfalo/metabolismo , Clonagem Molecular/métodos , DNA Complementar/metabolismo , Gânglios Espinais/citologia , Regulação da Expressão Gênica/fisiologia , Glicoproteínas/metabolismo , Humanos , Imuno-Histoquímica/métodos , Hibridização In Situ/métodos , Macaca fascicularis , Camundongos , Dados de Sequência Molecular , Família Multigênica/genética , RNA Mensageiro/biossíntese , Receptores de Droga/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Alinhamento de Sequência , Especificidade da Espécie , Substância P/metabolismo , Fatores de Transcrição/classificação
13.
Brain Res Mol Brain Res ; 109(1-2): 18-33, 2002 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-12531512

RESUMO

We report here the isolation of a novel gene termed mGluR5R (mGluR5-related). The N-terminus of mGluR5R is highly similar to the extracellular domain of metabotropic glutamate receptor 5 (mGluR5) whereas the C-terminus bears similarity to the testis-specific gene, RNF18. mGluR5R is expressed in the human CNS in a coordinate fashion with mGluR5. Although the sequence suggests that mGluR5R may be a secreted glutamate binding protein, we found that when expressed in HEK293 cells it was membrane associated and not secreted. Furthermore, mGluR5R was incapable of binding the metabotropic glutamate receptor class I selective agonist, quisqualate. Although mGluR5R could not form disulfide-mediated covalent homodimers, it was able to form a homomeric complex, presumably through noncovalent interactions. mGluR5R also formed noncovalent heteromeric associations with an engineered construct of the extracellular domain of mGluR5 as well as with full-length mGluR5 and mGluR1alpha. The ability of mGluR5R to associate with mGluR1alpha and mGluR5 suggests that it may be a modulator of class I metabotropic glutamate receptor function.


Assuntos
Receptores de Glutamato Metabotrópico/genética , Receptores de Glutamato Metabotrópico/metabolismo , Sequência de Aminoácidos , Proteínas de Transporte/genética , Fracionamento Celular , Linhagem Celular , Sistema Nervoso Central/metabolismo , Meios de Cultivo Condicionados , Agonistas de Aminoácidos Excitatórios/metabolismo , Humanos , Substâncias Macromoleculares , Dados de Sequência Molecular , Ligação Proteica , Ácido Quisquálico/metabolismo , Receptor de Glutamato Metabotrópico 5 , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Alinhamento de Sequência
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