Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 98
Filtrar
1.
J Biol Chem ; 299(8): 105020, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37423300

RESUMO

Mammalian type opsin 5 (Opn5m), a UV-sensitive G protein-coupled receptor opsin highly conserved in vertebrates, would provide a common basis for UV sensing from lamprey to humans. However, G protein coupled with Opn5m remains controversial due to variations in assay conditions and the origin of Opn5m across different reports. Here, we examined Opn5m from diverse species using an aequorin luminescence assay and Gα-KO cell line. Beyond the commonly studied major Gα classes, Gαq, Gα11, Gα14, and Gα15 in the Gq class were individually investigated in this study, as they can drive distinct signaling pathways in addition to a canonical calcium response. UV light triggered a calcium response via all the tested Opn5m proteins in 293T cells, which was abolished by Gq-type Gα deletion and rescued by cotransfection with mouse and medaka Gq-type Gα proteins. Opn5m preferentially activated Gα14 and close relatives. Mutational analysis implicated specific regions, including α3-ß5 and αG-α4 loops, αG and α4 helices, and the extreme C terminus, in the preferential activation of Gα14 by Opn5m. FISH revealed co-expression of genes encoding Opn5m and Gα14 in the scleral cartilage of medaka and chicken eyes, supporting their physiological coupling. This suggests that the preferential activation of Gα14 by Opn5m is relevant for UV sensing in specific cell types.


Assuntos
Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP , Opsinas , Humanos , Camundongos , Animais , Opsinas/genética , Opsinas/metabolismo , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/genética , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Cálcio/metabolismo , Transdução de Sinais , Receptores Acoplados a Proteínas G/metabolismo , Opsinas de Bastonetes/metabolismo , Mamíferos/metabolismo
2.
J Biol Chem ; 299(7): 104899, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37295773

RESUMO

Opsins are universal photosensitive proteins in animals. Vertebrates have a variety of opsin genes for visual and non-visual photoreceptions. Analysis of the gene structures shows that most opsin genes have introns in their coding regions. However, teleosts exceptionally have several intron-less opsin genes that are presumed to have been duplicated by an RNA-based gene duplication mechanism, retroduplication. Among these retrogenes, we focused on the Opn4 (melanopsin) gene responsible for non-image-forming photoreception. Many teleosts have five Opn4 genes including one intron-less gene, which is speculated to have been formed from a parental intron-containing gene in the Actinopterygii. In this study, to reveal the evolutionary history of Opn4 genes, we analyzed them in teleost (zebrafish and medaka) and non-teleost (bichir, sturgeon, and gar) fishes. Our synteny analysis suggests that the intron-less Opn4 gene emerged by retroduplication after the branching of the bichir lineage. In addition, our biochemical and histochemical analyses showed that, in the teleost lineage, the newly acquired intron-less Opn4 gene became abundantly used without substantial changes in the molecular properties of the Opn4 protein. This stepwise evolutionary model of Opn4 genes is quite similar to that of rhodopsin genes in the Actinopterygii. The unique acquisition of rhodopsin and Opn4 retrogenes would have contributed to the diversification of the opsin gene repertoires in the Actinopterygii and the adaptation of teleosts to various aquatic environments.


Assuntos
Evolução Molecular , Peixes , Íntrons , Opsinas , Animais , Peixes/genética , Peixes/fisiologia , Opsinas/genética , Opsinas/metabolismo , Filogenia , Rodopsina/genética , Rodopsina/metabolismo , Peixe-Zebra/genética , Oryzias/genética , Sintenia/genética
3.
Cancer Sci ; 115(2): 660-671, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38130032

RESUMO

REV7 is a multifunctional protein implicated in various biological processes, including DNA damage response. REV7 expression in human cancer cells affects their sensitivity to DNA-damaging agents. In the present study, we investigated the significance of REV7 in pancreatic ductal adenocarcinoma (PDAC). REV7 expression was immunohistochemically examined in 92 resected PDAC specimens and 60 endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNAB) specimens of unresectable PDAC treated with platinum-based chemotherapy, and its association with clinicopathologic features was analyzed. Although REV7 expression was not significantly associated with the progression of primary tumors (T-factor and Stage) in either resected or unresectable PDAC, decreased levels of REV7 expression in EUS-FNAB specimens of unresectable PDAC were significantly associated with better outcomes of platinum-based chemotherapy and a favorable prognosis. REV7-deficient PDAC cell lines showed suppressed cell growth and enhanced sensitivity to cisplatin in vitro. Tumor-bearing mice generated using REV7-deficient PDAC cell lines also showed enhanced sensitivity to cisplatin in vivo. RNA sequencing analysis using WT and REV7-deficient PDAC cell lines revealed that REV7 inactivation promoted the downregulation of genes involved in the DNA repair and the upregulation of genes involved in apoptosis. Our results indicate that decreased expression of REV7 is associated with better outcomes of platinum-based chemotherapy in PDAC by suppressing the DNA damage response. It is also suggested that REV7 is a useful biomarker for predicting the outcome of platinum-based chemotherapy and the prognosis of unresectable PDAC and is a potential target for PDAC treatment.


Assuntos
Adenocarcinoma , Fenômenos Biológicos , Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Animais , Camundongos , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Platina/uso terapêutico , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Carcinoma Ductal Pancreático/tratamento farmacológico , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/metabolismo , Adenocarcinoma/tratamento farmacológico , Reparo do DNA/genética
4.
Eur Spine J ; 2024 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-38844588

RESUMO

PURPOSE: The purpose of the present study was to investigate the association between quantitatively assessed trunk extensor strength and gait-induced back pain (GIBP) in patients with adult spinal deformity (ASD). METHODS: Ninety-five patients with ASD aged ≥ 50 years who were admitted to our hospital between April 2018 and March 2023 were included in the study. GIBP was evaluated through a 6-minute walking test (6MWT), with GIBP being defined as the occurrence of back pain during the evaluation and inability to complete the test. The patients were divided into three groups: difficulty completing the 6MWT (Group 1), ability to complete the 6MWT with breaks (Group 2), and ability to complete the 6MWT without taking a break (Group 3). The main independent variable was trunk extensor strength, which was measured using a hand-held dynamometer. Ordered logistic regression analysis was conducted to assess the association between GIBP and trunk extensor strength while adjusting for basic characteristics and radiographic parameters as covariates. RESULTS: The numbers of patients with ASD included in each group were; 27 in Group 1 (28.4%), 31 in Group 2 (32.6%), and 37 in Group 3 (39.0%). An ordered logistic regression analysis adjusted for basic characteristics and radiographic parameters, trunk extensor strength was significantly associated with GIBP (odds ratios, 1.128; 95% confidence intervals, 1.025-1.242). CONCLUSIONS: The results of the present study strongly indicate that trunk extensor strength is a valuable factor associated with GIBP in patients with ASD.

5.
Cell Tissue Res ; 391(2): 249-267, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36418571

RESUMO

Eyes shut homolog (EYS) encodes a proteoglycan and the human mutation causes retinitis pigmentosa type 25 (RP25) with progressive retinal degeneration. RP25 most frequently affects autosomal recessive RP patients with many ethnic backgrounds. Although studies using RP models have facilitated the development of therapeutic medications, Eys has been lost in rodent model animals. Here we examined the roles for Eys in the maintenance of photoreceptor structure and function by generating eys-null medaka fish using the CRISPR-Cas9 system. Medaka EYS protein was present near the connecting cilium of wild-type photoreceptors, while it was absent from the eys-/- retina. The mutant larvae exhibited a reduced visual motor response compared with wild-type. In contrast to reported eys-deficient zebrafish at the similar stage, no retinal cell death was detected in the 8-month post-hatching (8-mph) medaka eys mutant. Immunohistochemistry showed a significant reduction in the length of cone outer segments (OSs), retention of OS proteins in the inner segments of photoreceptors, and abnormal filamentous actin network at the base of cone OSs in the mutant retina by 8 mph. Electron microscopy revealed aberrant structure of calyceal processes, numerous vesiculation and lamellar interruptions, and autophagosomes in the eys-mutant cone photoreceptors. In situ hybridization showed an autophagy component gene, gabarap, was ectopically expressed in the eys-null retina. These results suggest eys is required for regeneration of OS, especially of cone photoreceptors, and transport of OS proteins by regulating actin filaments. Enhanced autophagy may delay the progression of retinal degeneration when lacking EYS in the medaka retina.


Assuntos
Oryzias , Degeneração Retiniana , Animais , Humanos , Oryzias/genética , Oryzias/metabolismo , Degeneração Retiniana/genética , Peixe-Zebra/metabolismo , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Células Fotorreceptoras de Vertebrados/metabolismo , Mutação/genética , Opsinas/metabolismo , Autofagia/genética
6.
Eur Spine J ; 32(6): 1887-1894, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37039881

RESUMO

PURPOSE: The aim is to investigate whether a simple prone posture assessment test (P-test) at baseline can be predict the effectiveness of at least 3 months of physiotherapy for adults with structural spinal disorders. METHODS: Seventy-six adults (age 71.0 ± 7.1 years) with structural spinal disorders who visited our outpatient clinic and underwent physiotherapy, which included muscle strength and range of motion training was provided once a week for a minimum of 3 months, and where the load was adjusted individually by the physiotherapist. The P-test is performed with the subject lying on the bed in a prone position and is positive if no low back pain is seen and the abdomen touches the bed. The Oswestry Disability Index (ODI) was used to assess disability. The minimum clinically important difference (MCID) was set at 10% improvement of the ODI score. Logistic regression analysis was performed to investigate the association between baseline P-test and achievement of ODI-MCID. RESULTS: The study population characteristics were: Sagittal vertical axis 138.1 ± 73.2 mm; Pelvic tilt, 36.9 ± 9.8 degrees; Pelvic incidence minus lumbar lordosis, 45.3 ± 22.1 degrees; and maximum coronal Cobb angle, 21.3 ± 19.7 degrees. Logistic regression analysis showed that being positive on the P-test was associated with the achievement of ODI-MCID (Odds ratio, 8.381; 95% confidence interval, 2.487-35.257). CONCLUSIONS: This study found that our developed P-test was a useful predictor of achieving the ODI-MCID in a cohort of adults with structural spinal disorders receiving at least 3 months of physiotherapy.


Assuntos
Lordose , Dor Lombar , Fusão Vertebral , Humanos , Adulto , Recém-Nascido , Resultado do Tratamento , Qualidade de Vida , Dor Lombar/terapia , Postura , Estudos Retrospectivos
7.
Surg Today ; 53(4): 420-427, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35984520

RESUMO

PURPOSE: We investigated the utility of the open abdominal management (OA) technique for ruptured abdominal aortic aneurysm (rAAA). METHODS: Between January 2016 and August 2021, 33 patients underwent open surgery for rAAA at our institution. The patients were divided into OA (n = 12) and non-OA (n = 21) groups. We compared preoperative characteristics, operative data, and postoperative outcomes between the two groups. The intensive care unit management and abdominal wall closure statuses of the OA group were evaluated. RESULTS: The OA group included significantly more cases of a preoperative shock than the non-OA group. The operation time was also significantly longer in the OA group than in the non-OA group. The need for intraoperative fluids, amount of bleeding, and need for blood transfusion were significantly higher in the OA group than in the non-OA group. Negative pressure therapy (NPT) systems are useful in OA. In five of the six survivors in the OA group, abdominal closure was able to be achieved using components separation (CS) technique. CONCLUSIONS: NPT and the CS technique may increase the abdominal wall closure rate in rAAA surgery using OA and are expected to improve outcomes.


Assuntos
Aneurisma da Aorta Abdominal , Ruptura Aórtica , Implante de Prótese Vascular , Procedimentos Endovasculares , Humanos , Procedimentos Endovasculares/métodos , Japão , Abdome/cirurgia , Transfusão de Sangue , Ruptura Aórtica/cirurgia , Aneurisma da Aorta Abdominal/cirurgia , Resultado do Tratamento , Estudos Retrospectivos , Fatores de Risco
8.
J Orthop Sci ; 2023 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-38151394

RESUMO

BACKGROUNDS: Evaluation of gait posture using a three-dimensional motion analysis system (3DMAS) revealed that elderly patients with adult spinal deformity (ASD) experience pelvic anteversion while walking. The purpose of this study was to investigate the influence of changes in pelvic anteversion during gait on walking ability and physical function in patients with ASD. METHODS: Fifty-four patients with ASD aged 50 years or older who were admitted to our hospital between March 2016 and December 2021 were included in the study. The 6-min walking distance (6MWD) was used to evaluate walking ability, and trunk and hip extensor strength were measured to evaluate physical function in the subjects. The 3DMAS was used to measure the subject's changes in pelvic anteversion during gait. After measuring the changes in pelvic anteversion, the median value of the study subjects was calculated, according to which the subjects were divided into two groups (small anteversion [S] group, large anteversion [L] group). Walking ability and physical function were compared between the two groups. RESULTS: The number of subjects in each group was 27. Comparisons of walking ability and physical function between the groups revealed significant differences in 6MWD (S group, 333.6 ± 111.2 m; L group, 238.0 ± 106.3 m) and hip extensor strength (S group, 15.8 ± 3.8 kgf; L group, 13.4 ± 4.4 kgf). No significant differences regarding trunk extensor strength were observed between the groups (S group, 15.2 ± 4.0 kgf; L group, 12.9 ± 4.8 kgf). CONCLUSION: The results of the present study revealed that ASD patients with greater pelvic anteversion associated with walking have lower walking ability and physical function. These results suggest the importance of evaluating the posture of ASD patients not only by using radiographic findings but also by assessing movement, such as gait posture.

9.
Protein Expr Purif ; 195-196: 106096, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35460871

RESUMO

Plasmodium vivax ookinete surface protein, Pvs25, is a candidate for a transmission-blocking vaccine (TBV) for malaria. Pvs25 has four EGF-like domains containing 22 cysteine residues forming 11 intramolecular disulfide bonds, a structural feature that makes its recombinant protein expression difficult. In this study, we report the high expression of recombinant Pvs25 as a soluble form in silkworm, Bombyx mori. The Pvs25 protein was purified from hemolymphs of larvae and pupae by affinity chromatography. In the Pvs25 expressed by silkworm, no isoforms with inappropriate disulfide bonds were found, requiring no further purification step, which is necessary in the case of Pichia pastoris-based expression systems. The Pvs25 from silkworm was confirmed to be molecularly uniform by sodium dodecyl sulfate gel electrophoresis and size-exclusion chromatography. To examine the immunogenicity, the Pvs25 from B. mori was administered to BALB/c mice subcutaneously with oil adjuvant. The Pvs25 produced by silkworm induced potent and robust immune responses, and the induced antisera correctly recognized P. vivax ookinetes in vitro, demonstrating the potency of Pvs25 from silkworm as a candidate for a malaria TBV. To the best of our knowledge, this is the first study to construct a system for mass-producing malaria TBV antigens using silkworm.


Assuntos
Bombyx , Vacinas Antimaláricas , Malária Vivax , Animais , Antígenos de Protozoários/genética , Antígenos de Superfície , Bombyx/genética , Dissulfetos , Vacinas Antimaláricas/genética , Malária Vivax/prevenção & controle , Camundongos , Plasmodium vivax/genética
10.
BMC Vet Res ; 17(1): 319, 2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34592989

RESUMO

BACKGROUND: Lobular dissecting hepatitis (LDH) is a rare form of canine liver cirrhosis that may be accompanied by portal hypertension in American Cocker Spaniels. In human patients with liver cirrhosis, portal vein thrombosis (PVT) is a common complication. However, PVT has not been reported in dogs with LDH. Herein, we describe the long-term follow-up of PVT in an American Cocker Spaniel with LDH. CASE PRESENTATION: An 8-year-old neutered male American Cocker Spaniel presented with a 1-month history of severe abdominal effusion. The dog was histopathologically diagnosed with LDH and treated with low-dose prednisolone on day 14. On day 115, computed tomography angiography (CTA) confirmed the presence of a thrombus in the portal vein. Therefore, the dog was subcutaneously administered with the anticoagulant dalteparin, and low-dose prednisolone was continued. As a follow-up for PVT, CTA examinations were performed on days 207, 515, 886, and 1168, and the dog's antithrombin and D-dimer levels were measured. Following anticoagulant therapy, the dog was confirmed to have gradually increased antithrombin activity and decreased D-dimer concentrations. In addition, although the thrombus was confirmed to be in the same area of the portal vein system by CTA, atrophy and increased CT values due to organization were observed during the follow-up period. The dog's condition remained stable without clinical signs until day 1112 when it developed hepatic encephalopathy. The dog died on day 1208. On postmortem examination, histopathologically, the liver showed marked bile duct hyperplasia and fibrosis with chronic thrombus in the portal vein. CONCLUSIONS: This case demonstrated that low-dose glucocorticoid combined with dalteparin allowed long-term follow-up of PVT in an American Cocker Spaniel with LDH.


Assuntos
Dalteparina/uso terapêutico , Hepatite/complicações , Veia Porta , Trombose Venosa/veterinária , Animais , Anticoagulantes/uso terapêutico , Angiografia por Tomografia Computadorizada , Doenças do Cão/tratamento farmacológico , Cães , Seguimentos , Cirrose Hepática/veterinária , Masculino , Prednisolona/uso terapêutico , Trombose Venosa/complicações , Trombose Venosa/diagnóstico por imagem
11.
Cell Tissue Res ; 379(1): 157-167, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31673758

RESUMO

REIC (reduced expression in immortalized cells) has been identified as a gene whose expression was reduced in immortalized cultured cells. The REIC gene is identical to Dickkopf-3 (Dkk3), which encodes a secreted glycoprotein belonging to the Dkk family. Previously, we showed that Dkk3 protein is present in the mouse adrenal medulla. However, its role in this tissue has not been elucidated. To explore it, we performed electron microscopic (EM) studies and RNA-sequencing (RNA-seq) analysis on Dkk3-null adrenal glands. EM studies showed that the number of dense core secretory vesicles were significantly reduced and empty vesicles were increased in the medulla endocrine cells. Quantitative PCR (qPCR) analysis showed relative expression levels of chromogranin A (Chga) and neuropeptide Y (Npy) were slightly but significantly reduced in the Dkk3-null adrenal glands. From the result of RNA-seq analysis as a parallel study, we selected three of the downregulated genes, uncoupled protein-1 (Ucp1), growth arrest and DNA-damage-inducible 45 gamma (Gadd45g), and Junb with regard to the estimated expression levels. In situ hybridization confirmed that these genes were regionally expressed in the adrenal gland. However, expression levels of these three genes were not consistent as revealed by qPCR. Thus, Dkk3 maintains the integrity of secreting vesicles in mouse adrenal medulla by regulating the expression of Chga and Npy.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/fisiologia , Medula Suprarrenal/fisiologia , Vesículas Secretórias/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Medula Suprarrenal/citologia , Medula Suprarrenal/ultraestrutura , Animais , Cromogranina A/metabolismo , Regulação para Baixo , Feminino , Hibridização In Situ , Camundongos , Camundongos Knockout , Neuropeptídeo Y/metabolismo , RNA Mensageiro , RNA-Seq , Vesículas Secretórias/ultraestrutura , Transcriptoma
12.
Eur J Orthop Surg Traumatol ; 30(2): 207-213, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31595359

RESUMO

PURPOSE: This study was to evaluate clinical outcomes using a patient-oriented test that scores health-related quality of life (HRQOL) for patients after minimally invasive surgery using microendoscopic discectomy (MED) for lumbar disc hernia. Few studies regarding MED in terms of disease-specific quality of life measures using Japanese Orthopaedic Association Back Pain Evaluation Questionnaire (JOABPEQ) have been published. METHODS: Retrospective analysis of the surgical and clinical outcomes with regard to reducing pain and improving the functional status for 31 patients who underwent MED for lumbar disc hernia was conducted. These patients were evaluated at 3-year follow-up. The evaluations were based on a visual analogue scale (VAS), the Japanese Orthopaedic Association (JOA) scoring system, and the JOABPEQ, which is an objective, patient-oriented test that assesses HRQOL in patients with lumbar disorders. RESULTS: A low rate of improvement was seen only in mental health until 1 year, the low rate of improvement in mental health and was independently correlated with body mass index (BMI), pre-operative scores on the Brief Scale for Psychiatric problems in Orthopaedic Patients (BS-POP), and scores on the BS-POP at 12 months post-operatively. CONCLUSIONS: All categories of VAS, JOA scores, and all domains of JOABPEQ were significantly higher over 3 years than those obtained pre-operatively. But only mental health domain showed mild improvement until 1 year. Moreover, BMI showed a negative correlation with improvements in the mental health domain post-operatively. As patients may be mentally exhausted from lumbar disc herniation, pre-operative mental health may be improved by surgical treatment.


Assuntos
Discotomia/métodos , Deslocamento do Disco Intervertebral/cirurgia , Dor Lombar/psicologia , Dor Lombar/cirurgia , Qualidade de Vida , Inquéritos e Questionários , Adulto , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Endoscopia , Feminino , Seguimentos , Humanos , Deslocamento do Disco Intervertebral/complicações , Dor Lombar/etiologia , Vértebras Lombares , Masculino , Saúde Mental , Pessoa de Meia-Idade , Núcleo Pulposo , Medição da Dor/métodos , Escalas de Graduação Psiquiátrica , Estudos Retrospectivos
13.
Immunogenetics ; 71(10): 635-645, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31745606

RESUMO

It has become anticipated that regenerative medicine will extend into the field of veterinary medicine as new treatments for various disorders. Although the use of allogeneic stem cells for tissue regeneration is more attractive than that of autologous cells in emergencies, the therapeutic potential of allogeneic transplantation is often limited by allo-immune responses inducing graft rejection. Therefore, a methodology for quantifying and monitoring alloreactive T cells is necessary for evaluating allo-immune responses. The mixed lymphocyte reaction (MLR) is widely used to evaluate T cell alloreactivity. In human, flow cytometric MLR with carboxyfluorescein diacetate succinimidyl ester has been established and used as a more useful assay than conventional MLR with radioisotope labeling. However, the available information about alloreactivity based on the differences of dog major histocompatibility complex (MHC) (dog leukocyte antigen, DLA) is quite limited in dog. In this paper, we describe our established flow cytometric MLR method that can quantify the T cell alloreactivity while distinguishing cell phenotypes in dog, and T cell alloreactivity among DLA-type matched pairs was significantly lower than DLA-mismatched pairs, suggesting that our developed flow cytometric MLR method is useful for quantifying T cell alloreactivity. In addition, we demonstrated the advantage of DLA homozygous cells as a donor (stimulator) for allogeneic transplantation. We also elucidated that the frequency of alloreactive T cell precursors was almost the same as that of mouse and human (1-10%). To our knowledge, this is the first report to focus on the degree of allo-immune responses in dog based on the differences of DLA polymorphisms.


Assuntos
Citometria de Fluxo/métodos , Histocompatibilidade , Teste de Cultura Mista de Linfócitos/métodos , Complexo Principal de Histocompatibilidade/genética , Complexo Principal de Histocompatibilidade/imunologia , Polimorfismo Genético , Linfócitos T/imunologia , Animais , Cães , Haplótipos , Ativação Linfocitária/imunologia
14.
Langmuir ; 33(15): 3680-3684, 2017 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-28351149

RESUMO

A novel type of energy donor-acceptor system on a clay surface has been prepared. The energy transfer between an energy-donating cationic pyrene derivative (An-Py2+) and an energy-accepting tris(bipyridine)ruthenium complex (Ru2+) on the clay surface was investigated using absorption, emission, and lifetime measurements. An obvious energy transfer was observed, and one Ru2+ molecule quenched the emission from five molecules of An-Py2+ with an emission quenching efficiency of 85% on the clay surface. This suggests that the light energies absorbed by five of the An-Py2+ molecules were accumulated in the one Ru2+ molecule. Near-quantitative emission quenching was observed for stoichiometric amounts of An-Py2+ and Ru2+. The apparent quenching rate constant is approximately 1017 L mol-1 s-1, and thus the quenching rate constant is 107-108 times higher than the diffusion rate constant in a homogeneous solution.

15.
Langmuir ; 31(1): 27-31, 2015 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-25540843

RESUMO

Tetra-cationic pyrene derivative (Py(4+)) and tris(bipyridine)ruthenium(II) (Ru(2+)) were hybridized onto the surface of a synthesized clay. We observed the remarkable stimulation of excited Py(4+) emission quenching on the clay surface, with a very large apparent quenching rate constant (kq = 7.4 ± 0.7 × 10(15) L mol(-1) s(-1)).

17.
Biochemistry ; 53(27): 4420-5, 2014 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-24960425

RESUMO

Rhodopsin is a G protein-coupled receptor specialized for photoreception and contains a light-absorbing chromophore retinal that binds to the lysine residue of opsin through a protonated Schiff base linkage. Light converts rhodopsin to an equilibrium mixture of the active state metarhodopsin II (MII) and its precursor, metarhodopsin I (MI), which have deprotonated and protonated Schiff base chromophores, respectively. This equilibrium was thought to depend on the pKa of not the Schiff base chromophore but glutamic acid E134 in the highly conserved E/DRY triad in helix III. We performed mutational analyses of E134 and nearby residues to examine whether the equilibrium is really dependent on the pKa of E134 and to obtain clues about the contribution of E134 to the G protein activation characteristics of rhodopsin. All the single mutants at position 134 except for E134D lost the characteristic pH-dependent equilibrium, indicating that the carboxyl group of E134 is responsible for the equilibrium. Interestingly, mutation at position 134 caused little change in the MI or MII spectra or G protein activation efficiency of MII, while it caused a shift of the MI-MII equilibrium. The mutants containing hydrophobic or amide-containing residues at position 134 formed an equilibrium in favor of MII, resulting in an increase in light-induced G protein activation efficiency. On the other hand, the wild type exhibited an opsin activity lower than those of the mutants, which exhibited reasonable light-dependent activities. These results strongly suggest that the evolutionary significance of E134 is not an increase in G protein activity but rather suppression of the opsin activity.


Assuntos
Ácido Glutâmico/metabolismo , Rodopsina/metabolismo , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Domínio Catalítico , Bovinos , Sequência Conservada , Células HEK293 , Humanos , Concentração de Íons de Hidrogênio , Mutação , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rodopsina/genética , Transducina/metabolismo
18.
Respirology ; 19(2): 239-245, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25198924

RESUMO

BACKGROUND AND OBJECTIVE: The results of studies examining the outcome and the factors predicting prognosis in combined pulmonary fibrosis and emphysema (CPFE) have so far been contradictory. Our objective was to determine prognosis and the prognostic factors for CPFE. METHODS: Of 108 consecutive idiopathic pulmonary fibrosis (IPF) patients admitted to our hospital, 46 were diagnosed as having CPFE and 62 patients diagnosed as having IPF alone. We retrospectively compared the clinical features between these two groups. RESULTS: Annual increase in estimated systolic pulmonary arterial pressure (esPAP) was significantly greater in CPFE patients, and survival time was significantly lower. Moreover, the prognosis was unfavourable regardless of the presence of lung cancer. The multivariate Cox proportional hazard regression model showed that predictive factors were an increase in the modified Medical Research Council dyspnoea score and esPAP ≥ 30.4 mm Hg. We classified patients into the following four groups: CPFE with high esPAP (esPAP ≥ 30.4 mm Hg), CPFE with normal esPAP (esPAP < 30.4 mm Hg), IPF alone with high esPAP and IPF alone with normal esPAP. Survival in the CPFE with high esPAP group was significantly worse than that in the other three subgroups. Furthermore, CPFE with the paraseptal type of emphysema and high esPAP had the worst prognosis. CONCLUSIONS: This study demonstrated that the prognosis of CPFE is significantly worse than that of IPF alone. In particular, CPFE with paraseptal emphysema associated with high esPAP has an extremely poor prognosis.


Assuntos
Fibrose Pulmonar Idiopática/complicações , Enfisema Pulmonar/etiologia , Idoso , Progressão da Doença , Feminino , Humanos , Fibrose Pulmonar Idiopática/diagnóstico , Masculino , Prognóstico , Modelos de Riscos Proporcionais , Enfisema Pulmonar/diagnóstico , Estudos Retrospectivos
19.
Beilstein J Org Chem ; 10: 1504-12, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25161707

RESUMO

Glycosyl-[60]fullerenes were first used as decontaminants against ricin, a lactose recognition proteotoxin in the Ricinus communis family. A fullerene glycoconjugate carrying two lactose units was synthesized by a [3 + 2] cycloaddition reaction between C60 and the azide group in 6-azidohexyl ß-lactoside per-O-acetate. A colloidal aqueous solution with brown color was prepared from deprotected bis(lactosyl)-C60 and was found stable for more than 6 months keeping its red color. Upon mixing with an aqueous solution of Ricinus communis agglutinin (RCA120), the colloidal solution soon caused precipitations, while becoming colorless and transparent. In contrast, a solution of concanavalin A (Con A) caused no apparent change, indicating that the precipitation was caused specifically by carbohydrate-protein interactions. This notable phenomenon was quantified by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the results were discussed in terms of detection and decontamination of the deadly biological toxin in the Ricinus communis family.

20.
J Mol Biol ; 436(5): 168319, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-37865286

RESUMO

Animal opsin is a G-protein coupled receptor (GPCR) and binds retinal as a chromophore to form a photopigment. The Opsin 5 (Opn5) group within the animal opsin family comprises a diverse array of related proteins, such as Opn5m, a protein conserved across all vertebrate lineages including mammals, and other members like Opn5L1 and Opn5L2 found in non-mammalian vertebrate genomes, and Opn6 found in non-therian vertebrate genomes, along with Opn5 homologs present in invertebrates. Although these proteins collectively constitute a single clade within the molecular phylogenetic tree of animal opsins, they exhibit markedly distinct molecular characteristics in areas such as retinal binding properties, photoreaction, and G-protein coupling specificity. Based on their molecular features, they are believed to play a significant role in physiological functions. However, our understanding of their precise physiological functions and molecular characteristics is still developing and only partially realized. Furthermore, their unique molecular characteristics of Opn5-related proteins suggest a high potential for their use as optogenetic tools through more specialized manipulations. This review intends to encapsulate our current understanding of Opn5, discuss potential manipulations of its molecular attributes, and delve into its prospective utility in the burgeoning field of animal opsin optogenetics.


Assuntos
Opsinas , Optogenética , Receptores Acoplados a Proteínas G , Animais , Opsinas/química , Opsinas/classificação , Opsinas/genética , Filogenia , Receptores Acoplados a Proteínas G/química , Receptores Acoplados a Proteínas G/classificação , Receptores Acoplados a Proteínas G/genética , Sequência Conservada
SELEÇÃO DE REFERÊNCIAS
Detalhe da pesquisa