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1.
Clin Chem Lab Med ; 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38753471

RESUMO

OBJECTIVES: Even in the current era of hematology analyzer automation and peripheral equipment, quality control sample measurement remains a manual task, leading to variability in quality control data and increased workload. In this study, we evaluated the performance of quality control measurement using the BT-50 Transportation Unit (BT-50, Sysmex, Kobe, Japan), equipped with a scheduled automatic quality control function, to ensure measurement accuracy and streamline the workflow of hematology testing. METHODS: We evaluated the automatic measurement performance of quality control samples using the BT-50 for six representative blood test parameters: WBC (white blood cell), RBC (red blood cell), HGB (hemoglobin), HCT (hematocrit), PLT (platelet), and RET% (reticulocyte percent). We evaluated the equivalence and compared measurement accuracy between the BT-50 and the manual method. We then compared the variability to other laboratories and confirmed the stability of quality control samples. We also evaluated changes in workflow and staff resources before and after the introduction of the BT-50. RESULTS: The quality control measurement results for the BT-50 and the manual method were found to be equivalent for all six parameters. The variability measured by the BT-50 was lower for some parameters compared to the manual method. Furthermore, the workflow was streamlined by reducing manual processes, resulting in increased efficiency. CONCLUSIONS: We confirmed the performance of quality control measurements using the schedule function of the BT-50. Introducing the BT-50 reduced the operator's workload, improved operational efficiency, and promoted the standardization of quality control measurements.

2.
Eur J Cardiothorac Surg ; 65(4)2024 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-38489836

RESUMO

OBJECTIVES: After staged reconstruction for hypoplastic left heart syndrome, the neoaortic root tends to dilate, and the incidence of significant neoaortic valve insufficiency increases with time. This study aimed to evaluate the mid-term outcomes of the neoaortic root geometries and valve function after chimney reconstruction in the Norwood procedure. METHODS: Between 2013 and 2021, 20 consecutive patients who underwent chimney reconstruction during the Norwood procedure for hypoplastic left heart syndrome and its variants in our institution were enrolled. The actual diameters of the following points were measured, and Z-scores were calculated based on the normal aortic root geometries using the long axis view of echocardiography at the pre-Norwood stage and the lateral view of angiography at pre-Glenn, pre-Fontan, post-Fontan and follow-up (age 5-6 years) stages: neoaortic valve annulus; sinus of Valsalva; sinotubular junction; and ascending aorta just proximal to the anastomosis to the aortic arch. The degree of neoaortic valve regurgitation was evaluated by echocardiography at each stage. RESULTS: The median follow-up period was 3.9 years. Neoaortic roots after chimney reconstruction were spared from progressive dilation over time. With growth, the conical configuration of the neoaortic roots was preserved without geometrical distortion. The Z-scores of the annulus, sinus of Valsalva, sinotubular junction and ascending aorta ranged roughly from 4 to 6, 4 to 6, 2 to 4 and 0 to 2, respectively. All neoaortic valves at each stage had mild or no regurgitation. CONCLUSIONS: Chimney reconstruction prevented neoaortic root dilation and avoided significant neoaortic valve regurgitation in the mid-term. These neoaortic dimensions with smooth flow profiles in the neoaorta after chimney reconstruction may have contributed to the current results. Further studies are needed to clarify the long-term outcomes.


Assuntos
Insuficiência da Valva Aórtica , Síndrome do Coração Esquerdo Hipoplásico , Procedimentos de Norwood , Humanos , Pré-Escolar , Criança , Síndrome do Coração Esquerdo Hipoplásico/cirurgia , Resultado do Tratamento , Procedimentos de Norwood/efeitos adversos , Aorta/cirurgia , Ecocardiografia , Insuficiência da Valva Aórtica/cirurgia , Insuficiência da Valva Aórtica/etiologia , Seguimentos , Estudos Retrospectivos
4.
J Artif Organs ; 2023 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-37227546

RESUMO

PURPOSE: A narrow terminal aorta is a risk factor for endograft occlusion after endovascular aneurysm repair. To minimize limb complications, we used Gore Excluder legs positioned side-by-side at the terminal aorta. We investigated the outcomes of our strategy for endovascular aneurysm repair in patients with a narrow terminal aorta. METHODS: We enrolled 61 patients who underwent endovascular aneurysm repair with a narrow terminal aorta (defined as < 18 mm in diameter) from April 2013 to October 2021. The standard procedure involves complete treatment with the Gore Excluder device. When other types of main body endografts were used, they were deployed proximal to the terminal aorta, and we used the Gore Excluder leg device in the bilateral limbs. Postoperatively, the intraluminal diameter of the legs at the terminal aorta was measured to assess the configuration. RESULTS: During the follow-up period (mean: 2.7 ± 2.0 years), there were no aorta-related deaths, endograft occlusions, or leg-related re-interventions. There were no significant differences between the pre- and postoperative ankle-brachial pressure index values in the dominant and non-dominant legs (p = 0.44 and p = 0.17, respectively). Postoperatively, the mean difference rate (defined as [dominant leg diameter-non-dominant leg diameter]/terminal aorta diameter) was 7.5 ± 7.1%. The difference rate was not significantly correlated with the terminal aortic diameter, calcification thickness, or circumferential calcification (r = 0.16, p = 0.22; r = 0.07, p = 0.59; and r = - 0.07, p = 0.61, respectively). CONCLUSIONS: Side-by-side deployment of Gore Excluder legs produces acceptable outcomes for endovascular aneurysm repair with a narrow terminal aorta. The endograft expansion at the terminal aorta is tolerable without influencing calcification distribution.

5.
Ann Thorac Surg ; 115(1): e11-e13, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-35259392

RESUMO

In pediatric cardiovascular surgery, the autologous pericardium is useful for pulmonary artery augmentation. However, in some cases, the autologous pericardium may not be available for various reasons. Since 2014, we have developed tissue-engineered vascular grafts (TEVGs) and obtained good results. We report a successful case of multiple pulmonary artery plasty with TEVGs for pulmonary atresia/ventricular septal defect with major aortopulmonary collateral arteries. TEVGs are useful alternatives to autologous pericardium in pediatric cardiovascular operations that often require multistage procedures.


Assuntos
Cardiopatias Congênitas , Comunicação Interventricular , Atresia Pulmonar , Criança , Humanos , Lactente , Artéria Pulmonar/cirurgia , Prótese Vascular , Comunicação Interventricular/cirurgia , Atresia Pulmonar/cirurgia , Circulação Colateral
6.
Front Sports Act Living ; 4: 941042, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35935065

RESUMO

To evaluate competitive ballroom dancing, one of the official events of the World Games, spin movements used by advanced dancers who ranked high in actual competition may be employed. The purpose of this study was to investigate the kinematic characteristics of spin movements in competitive ballroom dancing performed by world champion couples, especially holding posture and lower limb movements. A champion couple and 13 national-level competitive ballroom dancers as the control group participated in this study. An inertial measurement unit (IMU) system (MVN Link; Xsens, Netherlands) consisting of 17 IMUs, attached to feet, shanks, thighs, pelvis, sternum, head, upper arms, forearms, and hands of the dancers, was used to obtain three-dimensional kinematic data at 240 Hz. The overall trend was that the actual and normalized stride lengths of the champion male and female dancers tended to be longer than those of the male and female dancers in the control group. Further, large differences were observed in the pelvic and rib cage segments movements, and the relative angle of the rib cage segment to the pelvic segment. The champion male dancer started to move from the pelvic segment, whereas the champion female dancer and the national-level top male and female dancers started to move from the rib cage segment. During the spin movement, the champion male dancer was in a position where the rib cage segment was rotated to the left with respect to the pelvic segment, whereas the other dancers were in a position where the rib cage segment was rotated to the right. Although limited to technical aspects, these dance kinematic characteristics of the world champion couples and their differences from those of the control group dancers, will be helpful to competitive ballroom dancers and coaches in their daily practice.

7.
Asian Cardiovasc Thorac Ann ; 30(8): 931-934, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35821584

RESUMO

Left ventricular aneurysms are rarely encountered in pediatric patients. A 4-year-old boy was diagnosed with severe mitral regurgitation and a posterior left ventricular aneurysm associated with a viral infection. The aneurysm was surgically resected and plicated longitudinally with a combination of an interrupted mattress and continuous over-and-over sutures with an outer felt reinforcement. The mitral regurgitation was reduced to a trivial degree postoperatively. The patient's postoperative recovery was good. He was discharged 31 days after surgery without cardiac dysfunction or lethal arrhythmia. Considering the location and spread of the aneurysm, an appropriate surgical procedure should be adopted for pediatric patients.


Assuntos
Procedimentos Cirúrgicos Cardíacos , Aneurisma Cardíaco , Insuficiência da Valva Mitral , Procedimentos Cirúrgicos Cardíacos/métodos , Criança , Pré-Escolar , Aneurisma Cardíaco/complicações , Aneurisma Cardíaco/diagnóstico por imagem , Aneurisma Cardíaco/cirurgia , Ventrículos do Coração/diagnóstico por imagem , Ventrículos do Coração/cirurgia , Humanos , Masculino , Insuficiência da Valva Mitral/diagnóstico por imagem , Insuficiência da Valva Mitral/etiologia , Insuficiência da Valva Mitral/cirurgia , Resultado do Tratamento
8.
J Dance Med Sci ; 24(4): 168-174, 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-33218370

RESUMO

Competitive dance, also known as DanceSport, is one of the official sports of the World Games. The most salient characteristic of ballroom dance is the closed-hold position, during which the upper body segments of partner-dancers are linked. This study aimed to investigate partnering effects on joint motion ranges of the lower extremity and step lengths during the waltz in 13 national level competitive dance couples and a world champion couple. A Xsens MVN system was used to record movement at 240 Hz. Solo and pair conditions were examined. Compared with the highly skilled couples, the world champion couple demonstrated superior dance skills for generating the first step length in the pair condition of the waltz. This was particularly evident in the step length and joint motion range of the champion female dancer.


Assuntos
Desempenho Atlético/fisiologia , Comportamento Competitivo/fisiologia , Dança/fisiologia , Resistência Física/fisiologia , Amplitude de Movimento Articular/fisiologia , Antropometria/métodos , Feminino , Humanos , Masculino , Força Muscular/fisiologia
9.
JAMA Ophthalmol ; 135(5): 478-482, 2017 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-28358967

RESUMO

Importance: Ocular inflammation is occasionally observed after vaccinations, and most of them resolve without permanent visual disturbances. However, there are some rare cases of severe ocular complications following vaccinations. Objective: To report the findings in an infant boy who developed an acute loss of vision bilaterally after Haemophilus influenzae type b, Pneumococcal conjugate vaccination, and measles and rubella vaccination. His vision did not recover. Design, Setting, and Participant: A retrospective review of the medical records of a 13-month-old Japanese boy. Main Outcomes and Measures: Fundus and fluorescein angiographic findings, ultrasonographic and optical coherence tomographic images, and electroretinographic findings. Results: A healthy 13-month-old boy had an acute loss of vision in both eyes 31 days after Haemophilus influenzae type b and Pneumococcal conjugate vaccinations and 24 days after a measles and rubella vaccination. He also developed a common cold 10 days before the vision loss. Ultrasonography showed an exudative retinal detachment 1 day after the onset of the visual reduction; however, his fundi appeared normal 4 days later. His eyes did not pursue objects, and pupillary light reflexes were not present. No signs of anterior uveitis were noted. He was treated with corticosteroids, but his vision did not improve. The retinal vessels gradually attenuated, and diffuse small white punctate lesions appeared in the deep retina. Optical coherence tomography showed a thinner outer nuclear layer and an absent ellipsoid zone. The electroretinograms were nonrecordable. These findings suggested a severe impairment of the photoreceptors, especially their outer segments. Western blot analysis of the patient's sera detected an antibody against recoverin, a calcium-binding protein of photoreceptors. Conclusions and Relevance: We hypothesize that an infection induced severe chorioretinitis with an exudative retinal detachment, which then produced an autoantibody against recoverin. The autoantibody then altered the function of the photoreceptors very rapidly. The initial infection may have been caused by the measles and rubella vaccination. However, because to our knowledge this has not been reported previously, the visual loss after the vaccinations may have been an extremely rare event that was coincidental or may have been related to the vaccination.


Assuntos
Vacina contra Sarampo/efeitos adversos , Células Fotorreceptoras/patologia , Degeneração Retiniana/etiologia , Vacina contra Rubéola/efeitos adversos , Vacinação/efeitos adversos , Doença Aguda , Eletrorretinografia , Angiofluoresceinografia , Fundo de Olho , Humanos , Lactente , Masculino , Degeneração Retiniana/diagnóstico , Tomografia de Coerência Óptica/métodos
10.
Clin Chem Lab Med ; 53(2): 281-9, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25153399

RESUMO

BACKGROUND: The XN-Series (Sysmex, Kobe, Japan) have been equipped with the automated digital cell imaging analyzer DI-60, which provides complete automation of the sample processing with automated complete blood counts (CBC), slide making/staining, and digital scanning with cell pre-classification. The aim of this study was to evaluate the efficacy of the XN-Series as an integrated blood cell analysis system. METHODS: White blood cell (WBC) morphological analysis by the DI-60 was evaluated using 232 blood samples from patients. Routine analysis of a total of 2000 blood samples has been performed to evaluate the processing ability of the XN-Series connected to the DI-60. RESULTS: The overall analysis accuracy of pre-classification of WBC by the DI-60 was 88.4%. Good correlation was observed between final results of the DI-60 analysis and manual differentiation with high sensitivity and specificity for blasts and immature granulocytes. The sample processing time of the XN-Series, from automated CBC to cell pre-classification, was 38±1 min/single run and 165±12 min/500 CBC samples run (slide preparation rate 15.6%) with no sample hold-up at the DI-60. CONCLUSIONS: The automated morphological analysis capability of the DI-60 has potential usefulness in the integrated automated hematology analysis system of XN-Series.


Assuntos
Automação , Testes Hematológicos , Leucócitos/patologia , Testes Hematológicos/instrumentação , Humanos , Contagem de Leucócitos/instrumentação
11.
J Equine Sci ; 22(1): 9-15, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-24833982

RESUMO

The aim of present study was to clarify the post-natal profile of follicle-stimulating hormone (FSH), luteinizing hormone (LH), immunoreactive (ir)-inhibin, progesterone, testosterone, and estradiol-17ß, and their relationships in Thoroughbred colts. Six hundred and thirty-six colts were used for the study. Single plasma samples from each animal were harvested from the blood drawn through jugular venipuncture. The subjects were born with high amounts of progesterone, testosterone, and estradiol-17ß, all of which dropped significantly and remained at lower levels till the end of 6 months. FSH decreased transiently after birth until day 12 and then gradually increased to peak at day 100 which then maintained in lesser levels towards the end of the studied period. LH was highest during birth which decreased until day 26 and then increased slowly to sub-birth levels up to day 90. Animals were born with high amounts of ir-inhibin. It dropped slowly and halved by day 20 and then decreased towards rest of the studied period. The increase in FSH is negatively correlated with the declining ir-inhibin levels. The early increase in FSH can be the indication of early post-natal maturation of the hypothalamic pituitary testicular axis that ultimately might be responsible for priming the testes for future development.

12.
J Equine Sci ; 22(2): 29-36, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-24833985

RESUMO

Changes in follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, immunoreactive(ir)-inhibin, testosterone, estradiol-17ß, and insulin-like growth factor (IGF)-I in Thoroughbred stallions along with changes in prolactin secretion in geldings were studied. The correlations of day-length with changes in the concentrations of these hormones were also studied. Five stallions and thirteen geldings were employed to draw blood samples in monthly basis and radioimmunoassay was performed to measure these hormones. All hormones showed a seasonal pattern, the levels being highest during the breeding season and lowest during the winter months. Most of the hormones were at their highest concentration during the month of April, the mid of spring in northern hemisphere. The concentration of circulating IGF-I also demonstrated seasonality, the peak lying on the month of April. The plasma concentration of prolactin also increased during the breeding season. This phenomenon was similar both in stallions and geldings although geldings had lower concentration than that of stallions. The changes in concentration of prolactin in stallions and geldings correlated more towards the day-length than towards the temperature. These results clearly indicate the seasonality of pituitary and gonadal hormones of Thoroughbred stallions, the activity being highest during the month of April and May of the breeding season.

13.
Masui ; 52(6): 631-5, 2003 Jun.
Artigo em Japonês | MEDLINE | ID: mdl-12854479

RESUMO

BACKGROUND: In pediatric renal transplantation, an adult kidney is transplanted to a small child and it may cause severe cardiovascular insufficiency. METHODS: The anesthetic management of 15 pediatric recipients younger than 10 years, undergoing living-related renal transplantation were analyzed retrospectively. RESULTS: Before the operation, 3 of 15 patients were receiving hemodialysis, and the others were receiving peritoneal dialysis. Isoflurane was used for the anesthetic maintenance in most of cases. Arterial and central venous pressures were monitored invasively in all cases. Pulmonary arterial catheters and transesophageal echocardiography were used in some patients with cardiac complications. For intraoperative fluid management, a total volume of 18 ml.kg-1.h-1 (mean) was infused, which included approximately 8 ml.kg-1.h-1 of crystalloid, 7 ml.kg-1.h-1 of fresh frozen plasma, and 2 ml.kg-1.h-1 of washed red blood cells. Dopamine 1-7 micrograms.kg-1.min-1 and human atrial natriuretic peptide (hANP) 0.05-0.1 microgram.kg-1.min-1 were administered as diuretics. None of 15 patients developed severe cardiovascular insufficiency of poor diuresis. CONCLUSIONS: Cardiovascular stability and good function of the transplanted kidney were maintained by appropriate monitoring and continuous intravenous infusion of dopamine and hANP. It is important to infuse enough volume of fluid and blood before recanalization.


Assuntos
Anestesia/métodos , Transplante de Rim/métodos , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Falência Renal Crônica/terapia , Doadores Vivos , Masculino , Diálise Peritoneal , Diálise Renal , Estudos Retrospectivos
14.
Transpl Immunol ; 11(2): 147-53, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12799197

RESUMO

Natural killer (NK) cells play an important role in xenograft rejection. The aim of this study was to evaluate the co-effect of human leukocyte antigen (HLA)-G1 expression and the remodeling of glycoantigens such as the alpha-Gal epitope, Galalpha1,3Galbeta1,4GlcNAc-R, by the introduction of glycosyltransferase genes related to NK cell-mediated direct cytotoxicity. Human peripheral blood mononuclear cells or an NK-like cell line, YT cells, was used as an effector and pig endothelial cells (PEC) as the target. A PEC transfectant with HLA-G1 was first prepared by the transfection of HLA-G1 and human beta2 microglobulin. Several new transfectants were then established by the transfection of glycosyltransferase to the HLA-G1 transfectant. The effect of HLA-G1 on NK cell-mediated PEC lysis was lower than that by the glycosyltransferases. Therefore, in the case of the co-transfectants except for HLA-G1+alpha2,6sialyltransferase, such as HLA-G1+N-acetylglucosaminyltransferase-III and HLA-G1+alpha1,2fucosyltransferase, the effect of HLA-G1 expression on NK-mediated killing appeared to be accounted for by the transfected glycosyltransferase activities and the reduced alpha-Gal expression on the cell surface. However, these transfectants showed significant reductions in direct NK cell-mediated cytotoxicity, compared with the single HLA-G1 transfectant. The results herein suggest that a combination of HLA-G1 and glycosyltransferases has considerable potential for the downregulation of NK cell-mediated cytolysis.


Assuntos
Citotoxicidade Imunológica , Células Endoteliais/patologia , Glicosiltransferases/imunologia , Antígenos HLA/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Células Matadoras Naturais/imunologia , Animais , Células Cultivadas , Regulação para Baixo , Células Endoteliais/imunologia , Citometria de Fluxo , Glicosiltransferases/genética , Antígenos HLA/genética , Antígenos HLA-G , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Suínos , Transfecção
15.
Biol Reprod ; 68(1): 328-35, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12493730

RESUMO

To clarify the source of inhibins in equine female fetuses, concentrations of immunoreactive (ir-) inhibin, inhibin pro-alphaC, and inhibin A in both fetal and maternal circulation and in fetal ovaries were measured. In addition, the localization of inhibin alpha and inhibin/activin beta(A), and beta(B) subunits and the expression of inhibin alpha(A) and inhibin/activin beta(A) subunit mRNA in fetal ovaries were investigated using immunohistochemistry and in situ hybridization. Concentrations of circulating ir-inhibin, inhibin pro-alphaC, and inhibin A were remarkably more elevated in the fetal than in the maternal circulation between Days 100 and 250 of gestation. Fetal ovaries contained large amounts of ir-inhibin, inhibin pro-alphaC, and inhibin A. In contrast, these inhibin forms were undetectable in both the maternal ovaries and placenta. The inhibin alpha and inhibin/activin beta(A) and beta(B) subunit proteins were localized to enlarged interstitial cells of the equine fetal ovary. Expression of inhibin alpha and inhibin/activin beta(A) subunit mRNAs were also observed in the interstitial cells. We conclude that the main source of large amounts of inhibins in fetal circulation is interstitial cells of fetal ovary and is not of maternal origin. Furthermore, these inhibins may play some important physiological roles in the development of gonads in the equine fetus.


Assuntos
Feto/metabolismo , Cavalos/fisiologia , Inibinas/metabolismo , Ovário/metabolismo , Animais , Feminino , Cavalos/anatomia & histologia , Subunidades beta de Inibinas/genética , Subunidades beta de Inibinas/metabolismo , Inibinas/genética , Troca Materno-Fetal , Tamanho do Órgão , Ovário/anatomia & histologia , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
16.
Transplantation ; 73(10): 1582-9, 2002 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-12042643

RESUMO

BACKGROUND: The inhibitory function of HLA class I molecules, HLA-G1 and HLA-E, on natural killer (NK) cell-mediated cytolysis has previously been reported. In this study, we report on a study of the effects of the co-expression of these molecules on the inhibition of NK cell-mediated cytolysis, using a newly constructed gene. METHODS: Complementary DNA (cDNA) of HLA-G (G1 and G3), HLA-E, and human beta2-microglobulin (hbeta2m) were prepared and transfected into swine endothelial cell (SEC) and Chinese hamster ovarian tumor (CHO) cell. The leader peptide sequences of HLA-G1 and HLA -E genes were changed to VMAPRTLFL or VMAPRTLVL, which corresponds to the original HLA-G1 and HLA-A2. The cell surface expression of the modified genes was evaluated by flow cytometry, and NK cell-mediated cytolysis by human peripheral blood mononuclear cells (PBMC) was assessed. RESULTS: The transfectant with the hbeta2m and HLA-G1 genes showed a clear expression of the HLA-G1 molecule and had an inhibitory effect on NK cell-mediated SEC lysis. Whereas neither the transfectant with the hbeta2m and HLA-E genes, nor that with the hbeta2m and HLA-G3 genes, expressed the HLA molecule on SEC, the transfectant with triple genes, hbeta2m, HLA-E, and HLA-G3, expressed the HLA-E molecule and also inhibited NK-mediated SEC lysis. Conversely, the modification of the leader sequence of the HLA-E gene successfully induced the expression of the HLA-E molecule on the SEC surface. Furthermore, the transfectant expressed both HLA-G1 and HLA-E molecules, thus efficiently enhancing the inhibition of NK-mediated SEC lysis. CONCLUSION: The co-expression of HLA-G1 and HLA-E molecules with the modified genes has potential for use in preventing xenograft rejection, as mediated by human NK cells.


Assuntos
Endotélio Vascular/imunologia , Regulação da Expressão Gênica , Antígenos HLA/genética , Antígenos de Histocompatibilidade Classe I/genética , Células Matadoras Naturais/imunologia , Sinais Direcionadores de Proteínas/genética , Animais , Anticorpos Monoclonais/farmacologia , Células CHO , Clonagem Molecular , Cricetinae , Citotoxicidade Imunológica/imunologia , Primers do DNA , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica/imunologia , Humanos , Neoplasias Ovarianas , Reação em Cadeia da Polimerase , Proteínas Recombinantes/imunologia , Suínos , Transfecção , Microglobulina beta-2/genética , Antígenos HLA-E
17.
J Biochem ; 131(4): 517-22, 2002 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11926988

RESUMO

Glycoprotein-3-sulfotransferase (GP3ST) is a key enzyme in downregulating the expression of Galalpha1,3Galbeta1,4GlcNAc-R (the alpha-Gal epitope), via enzymatic competition with an alpha1,3 galactosyltransferase (alpha1,3GT), such as alpha2,6 sialyltransferase (alpha2,6ST). In this study, we report the dominance of GP3ST over alpha1,3GT using transfected pig endothelial cell (PEC) lines. The introduction of the GP3ST gene into PEC suppresses its antigenicity with respect to normal human pooled serum (NHS), including the alpha-Gal epitope and the Hanganutziu-Deicher (H-D) antigen, and, in addition, reduces the susceptibility to NHS in complement-mediated cell lysis. Western and lectin blot analyses of the products of parental PEC and its transfectants indicated that proteins smaller than 66 kDa have a diminished reactivity with NHS and the IB4 lectin. The levels of the alpha-Gal epitope in neutral glycosphingolipids were also decreased in the GP3ST transfectants as detected in thin layer chromatography by immunostaining. These data indicate that GP3ST is very effective in reducing xenoepitope levels.


Assuntos
Antígenos Heterófilos/imunologia , Sulfotransferases/química , Sulfotransferases/metabolismo , Animais , Antígenos Heterófilos/metabolismo , Western Blotting , DNA Complementar/metabolismo , Regulação para Baixo , Endotélio/citologia , Epitopos , Citometria de Fluxo , Glicoesfingolipídeos/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Lectinas/metabolismo , Camundongos , Plasmídeos/metabolismo , Sialiltransferases/metabolismo , Suínos , Transfecção , Transplante Heterólogo , alfa-Galactosidase/metabolismo , beta-D-Galactosídeo alfa 2-6-Sialiltransferase
18.
Mol Reprod Dev ; 61(3): 302-11, 2002 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11835575

RESUMO

Porcine membrane cofactor protein (pMCP) is abundantly expressed throughout the body with particularly strong expression on the vascular endothelia. Previous studies demonstrated that the promoter of the pMCP gene induced efficient expression of a human complement regulatory protein, decay-accelerating factor (DAF; CD55), in transgenic mice. In the present study, we tried to produce transgenic pigs with two hybrid genes, 0.9/hDAF and 5.4/hDAF, which were composed of human DAF (hDAF) gene regulated under pMCP promoters of different lengths (0.9 and 5.4 kb). Five live founder transgenic pigs were obtained only with the 0.9/hDAF construct. Although, four founder pigs transmitted the transgene to the second generation, the transmission rates varied among founders. We examined the expression of hDAF in tissues of descendants of two lines (Dm1 and Dm4). Human DAF specific RNAs were confirmed by an RT-PCR analysis in all organs examined. Levels of hDAF protein in the organs from the descendants of Dm1 line were higher than those in the corresponding human organs as determined by enzyme-linked immunosorbent assay. Immunohistochemical studies showed that the tissue distribution of hDAF in the descendants of both lines was similar to that of endogenous pMCP. The expression level of hDAF on the vascular endothelial cells in Dm1 line was twice that on the corresponding human cells. We tested whether proinflammatory cytokines upregulate an efficiency of pMCP promoter on hDAF expression in transgenic pigs. Although the expression of hDAF on the human endothelial cells increased with a combination of cytokines, tumor necrosis factor alpha and interferon-gamma, no cytokine-induced upregulation was seen in the cells of transgenic pigs. The endothelial cells from transgenic pigs exhibited high resistance to the human serum-mediated cytolysis.


Assuntos
Antígenos CD/genética , Antígenos CD55/genética , Antígenos CD55/metabolismo , Regulação da Expressão Gênica , Glicoproteínas de Membrana/genética , Regiões Promotoras Genéticas/genética , Suínos/genética , Animais , Animais Geneticamente Modificados , Western Blotting , Células Cultivadas , Citocinas/farmacologia , Endotélio Vascular/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Masculino , Proteína Cofatora de Membrana , Especificidade de Órgãos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transgenes/genética
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