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1.
Toxicology ; : 153064, 2021 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-34890705

RESUMO

Drug-induced lens opacity has the potential to cause blindness and is of concern in drug development. Inhibition of cholesterol biosynthesis is one of the causes of lens opacity. Lens opacity is only observed after chronic administration in in vivo nonclinical studies in drug development. Thus, to save resources (e.g., time and cost) and to reduce burden on animals, it is required to develop in vitro evaluation systems that can predict and avoid the risk of lens opacity earlier and easier. In this study, we investigated whether rat lens explant cultures could be useful for the evaluation of drug-induced lens opacity via inhibition of cholesterol biosynthesis. Nineteen drugs, including statins, allylamine, thiocarbamate, azole, and morpholine, which inhibit cholesterol biosynthesis, as well as a negative control (acetaminophen, rosiglitazone and troglitazone), were used. Rat lens explants were treated with drugs for 13 days at concentrations close to IC50 values or higher against cholesterol biosynthesis, and lens opacity (severity and region) was evaluated. In most cases, region-specific lens opacity limited in the equator to posterior pole, as observed in vivo was observed at IC50 values or higher concentrations. The severity of opacity was likely to be related to the inhibitory potency toward cholesterol biosynthesis, concentration of drugs distributed in the lens, or time of exposure. Furthermore, GSH levels were also involved in the deterioration of lens opacity. In conclusion, we demonstrated that rat lens explant cultures can be useful to assess the potential drug-induced lens opacity associated with inhibition of cholesterol biosynthesis and to elucidate the mechanisms of lens opacity.

2.
Int Cancer Conf J ; 10(3): 254-258, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34221842

RESUMO

This is the case report of primary malignant melanoma (MM) of uterine cervix treated by immune checkpoint inhibitor: the Pembrolizumab. Despite the merge of the novel drugs that has been strikingly improving prognosis of MM, we still struggle treatment of MM of uterine cervix that has aggressive characteristics with unknown etiology. We present our case to contribute its rarity of the disease case report, the primary MM of the uterine cervix that had poor response to pembrolizumab and had OS of 6 months. The treatment ineffectiveness is mainly considered for mucosal MM of low tumor mutation burden and its unusual type of pathology. Accumulation of retrospective studies exclusively on cervical melanoma needs to be proceeded to investigate on characteristics between poor and long survival to establish standardized treatment.

3.
Hinyokika Kiyo ; 67(5): 205-209, 2021 May.
Artigo em Japonês | MEDLINE | ID: mdl-34126664

RESUMO

Lymphoepithelioma-like carcinoma (LELC) of the ureter is very rare and only 14 previous cases have been reported. Here, we report a case of LELC of the ureter. A 76-year-old woman was admitted to our hospital complaining of gross hematuria. Left ureteral cancer was suspected by the imaging examination, and laparoscopic left total nephroureterectomy was performed. Histopathological examination showed pure type of LELC in the ureter. She is alive without disease recurrence at fifteen months after surgery.


Assuntos
Carcinoma de Células Escamosas , Ureter , Neoplasias Ureterais , Idoso , Feminino , Humanos , Recidiva Local de Neoplasia , Nefroureterectomia , Ureter/diagnóstico por imagem , Ureter/cirurgia , Neoplasias Ureterais/diagnóstico por imagem , Neoplasias Ureterais/cirurgia
4.
FASEB J ; 34(7): 9480-9497, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32474967

RESUMO

In vitro mouse spermatogenesis using a classical organ culture method became possible by supplementing basal culture medium with only the product of bovine serum albumin purified by chromatography (AlbuMAX), which indicated that AlbuMAX contained every chemical factor necessary for mouse spermatogenesis. However, since the identity of these factors was unclear, improvements in culture media and our understanding of the nutritional and signal substances required for spermatogenesis were hindered. In the present study, chemically defined media (CDM) without AlbuMAX was used to evaluate each supplementary factor and their combinations for the induction of spermatogenesis. Similar to in vivo conditions, retinoic acid, triiodothyronine (T3 ), and testosterone (T) were needed. Based on differences in spermatogenic competence between AlbuMAX, fetal bovine serum, and adult bovine serum, we identified α-tocopherol, which strongly promoted spermatogenesis when combined with ascorbic acid and glutathione. Differences were also observed in the abilities of lipids extracted from AlbuMAX using two different methods to induce spermatogenesis. This led to the identification of lysophospholipids, particularly lysophosphatidylcholine, lysophosphatidic acid, and lysophosphatidylserine, as important molecules for spermatogenesis. New CDM formulated based on these results induced and promoted spermatogenesis as efficiently as AlbuMAX-containing medium. In vitro spermatogenesis with CDM may provide a unique experimental system for research on spermatogenesis that cannot be performed in in vivo experiments.


Assuntos
Antioxidantes/farmacologia , Lisofosfolipídeos/farmacologia , Técnicas de Cultura de Órgãos/métodos , Espermatogênese , Testículo/citologia , Vitaminas/farmacologia , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Testículo/efeitos dos fármacos , Testículo/metabolismo
6.
Reprod Med Biol ; 18(4): 362-369, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31607796

RESUMO

Purpose: Mouse in vitro spermatogenesis is possible with classical organ culture methods, by placing the testis tissue at the interphase between culture medium and air. In this condition, however, a tissue piece tends to round up to be compact, whose central region suffers from shortage of nutrients and oxygen. In this study, the authors improved the culture condition by spreading each tissue thin and flat, by which they were able to get better access to the oxygen and nutrients. Methods: Immature mouse testis tissues placed on agarose gel block were forced to spread flat by covering with a polydimethylsiloxane (PDMS) ceiling chip (PC chip). They were then cultured for weeks and evaluated by the transgene expression of Acr-Gfp, which reflects the progression of spermatogenesis. Results: Testis tissues covered with PC chip initiated and maintained spermatogenesis in its wider region than those without PC chip covering. Flow cytometric analysis demonstrated that the PC method yielded more numerous meiotic germ cells than those without PC. Immunohistochemical examination confirmed the authentic histological figure of spermatogenesis from spermatogonia up to round or elongating spermatids. Conclusions: The PC chip method is simple and effective to improve the efficiency of in vitro spermatogenesis in the organ culture system.

7.
Sci Rep ; 9(1): 20329, 2019 12 30.
Artigo em Inglês | MEDLINE | ID: mdl-31889132

RESUMO

Osteoarthritis (OA) is one of the world's most common degenerative diseases, but there is no disease-modifying treatment available. Previous studies have shown that prostaglandin E2 (PGE2) and PGE2 receptor 4 (EP4) are involved in OA pathogenesis; however, their roles are not fully understood. Here, we examined the efficacy of oral administration of KAG-308, an EP4-selective agonist, in surgically induced mouse knee OA. Cartilage degeneration and synovitis were significantly inhibited by the KAG-308 treatment. Chondrocyte hypertrophy and expression of tumor necrosis factor alpha (TNF) and matrix metalloproteinase 13 (Mmp13) in the synovium were suppressed in the KAG-308-treated mice. In cultured chondrocytes, hypertrophic differentiation was inhibited by KAG-308 and intranuclear translocation of histone deacetylase 4 (Hdac4) was enhanced. In cultured synoviocytes, lipopolysaccharide (LPS)-induced expression of TNF and Mmp13 was also suppressed by KAG-308. KAG-308 was detected in the synovium and cartilage of orally treated mice. TNF secretion from the synovia of KAG-308-treated mice was significantly lower than control mice. Thus, we conclude that oral administration of KAG-308 suppresses OA development through suppression of chondrocyte hypertrophy and synovitis. KAG-308 may be a potent candidate for OA drug development.


Assuntos
Anti-Inflamatórios/farmacologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Epoprostenol/análogos & derivados , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Receptores de Prostaglandina E Subtipo EP4/antagonistas & inibidores , Fator de Necrose Tumoral alfa/biossíntese , Administração Oral , Animais , Anti-Inflamatórios/administração & dosagem , Biópsia , Cartilagem/efeitos dos fármacos , Cartilagem/metabolismo , Cartilagem/patologia , AMP Cíclico , Modelos Animais de Doenças , Epoprostenol/administração & dosagem , Epoprostenol/farmacologia , Humanos , Camundongos , Osteoartrite do Joelho/tratamento farmacológico , Transporte Proteico , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Transdução de Sinais , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia
8.
Biotechnol Bioeng ; 115(12): 3030-3041, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30144353

RESUMO

Organ culture experiments can be hampered by central degeneration or necrosis due to the inadequate permeation of oxygen and nutrients, which deteriorates the function and growth of cultured tissues. In the current study, we aimed to overcome this limitation of organ culture through spreading the tissue two dimensionally on an agarose gel stand and molding into a disc shape by placing a ceiling of polydimethylsiloxane (PDMS) chip, which is highly oxygen permeable. By this, every part of the tissue can receive a sufficient supply of oxygen through PDMS as well as nutrients through the agarose gel below. This method not only prevented central necrosis of tissues, but also supported the tissue growth over time. In addition, such growth, as volume enlargement, could be easily measured. Under these conditions, we examined the effect of several factors on the growth of neonatal mouse testis, and found that follicle stimulating hormone (FSH) and insulin significantly promoted the growth. These results are in good agreement with previous in vivo reports. Notably, the growth achieved over 7 days in our in vitro system is almost comparable to, about 80% of, that observed in vivo. Thus, we successfully monitored the promotion of tissue growth beyond the limits of the conventional organ culture method. This extremely simple method could offer a unique platform to evaluate the growth as well as functional properties of organs, not only the testis but also others as well.


Assuntos
Técnicas de Cultura de Órgãos/instrumentação , Técnicas de Cultura de Órgãos/métodos , Testículo/citologia , Testículo/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Células Cultivadas , Dimetilpolisiloxanos/química , Dispositivos Lab-On-A-Chip , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Nylons/química , Células de Sertoli/citologia
9.
Biochem Biophys Res Commun ; 500(4): 885-891, 2018 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-29705697

RESUMO

In our previous study, we produced a microfluidic device (MFD) which successfully maintained spermatogenesis for over 6 months in mouse testis tissues loaded in the device. In the present study, we developed a new MFD, a monolayer device (ML-D) with a barrier structure consisting of pillars and slits, which is simpler in design and easier to make. This ML-D was also effective for inducing mouse spermatogenesis and maintained it for a longer period than the conventional culture method. In addition, we devised a way of introducing sample tissue into the device during its production, just before bonding the upper layer of polydimethylsiloxane (PDMS) and bottom glass slide. The tissue can obtain nutrients horizontally from the medium running beside it and oxygen vertically from above through PDMS. In addition, the glass slide set at the bottom improved the visibility of the sample tissue with an inverted microscope. When we took photos of cultured tissue of the Acr-Gfp transgenic mouse testis in ML-D sequentially every day, morphological changes of the acrosome during spermiogenesis were successfully recorded. The ML-D is simple in design and useful for culturing testis tissue for inducing and maintaining spermatogenesis with clearer visibility. Due to the new method of sample loading, tissues other than testis should also be applicable.


Assuntos
Desenho de Equipamento/instrumentação , Dispositivos Lab-On-A-Chip , Espermatogênese/genética , Espermatozoides/ultraestrutura , Testículo/citologia , Animais , Dimetilpolisiloxanos/química , Expressão Gênica , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Masculino , Camundongos , Camundongos Transgênicos , Espermatozoides/crescimento & desenvolvimento , Espermatozoides/metabolismo , Testículo/crescimento & desenvolvimento , Testículo/metabolismo , Técnicas de Cultura de Tecidos
10.
PLoS One ; 13(2): e0192884, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29432471

RESUMO

We previously reported the successful induction and completion of mouse spermatogenesis by culturing neonatal testis tissues. The culture medium consisted of α-minimum essential medium (α-MEM), supplemented with Knockout serum replacement (KSR) or AlbuMAX, neither of which were defined chemically. In this study, we formulated a chemically defined medium (CDM) that can induce mouse spermatogenesis under organ culture conditions. It was found that bovine serum albumin (BSA) purified through three different procedures had different effects on spermatogenesis. We also confirmed that retinoic acid (RA) played crucial roles in the onset of spermatogonial differentiation and meiotic initiation. The added lipids exhibited weak promoting effects on spermatogenesis. Lastly, luteinizing hormone (LH), follicle stimulating hormone (FSH), triiodothyronine (T3), and testosterone (T) combined together promoted spermatogenesis until round spermatid production. The CDM, however, was not able to produce elongated spermatids. It was also unable to induce spermatogenesis from the very early neonatal period, before 2 days postpartum, leaving certain factors necessary for spermatogenic induction in mice unidentified. Nonetheless, the present study provided important basic information on testis organ culture and spermatogenesis in vitro.


Assuntos
Técnicas de Cultura de Órgãos/métodos , Espermatogênese , Fatores Etários , Animais , Bovinos , Meios de Cultura/química , Hormônio Foliculoestimulante/farmacologia , Técnicas In Vitro , Lipídeos/farmacologia , Hormônio Luteinizante/farmacologia , Masculino , Meiose/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Camundongos Transgênicos , Soroalbumina Bovina/isolamento & purificação , Soroalbumina Bovina/farmacologia , Transdução de Sinais , Espermatogênese/efeitos dos fármacos , Espermatogênese/fisiologia , Espermatogônias/efeitos dos fármacos , Testículo/citologia , Testículo/efeitos dos fármacos , Testículo/fisiologia , Testosterona/farmacologia , Tretinoína/farmacologia , Tri-Iodotironina/farmacologia
11.
Sci Rep ; 7(1): 15459, 2017 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-29133858

RESUMO

Three-dimensional aggregation and organ culture methods are critical for recreating in vivo cellular phenomena outside the body. Previously, we used the conventional gas liquid interphase organ culture method to induce complete mouse spermatogenesis. After incorporating microfluidic systems, we achieved a significant increase in efficiency and duration of spermatogenesis. One of the major drawbacks preventing the popularization of microfluidics, however, is the use of a power-pump to generate medium flow. In this study, we produced a pumpless microfluidic device using hydrostatic pressure and a resistance circuit to facilitate slow, longer lasting medium flow. During three months of culture, results in induction and maintenance of spermatogenesis showed no difference between pumpless and pump-driven devices. Correspondingly, the spermatogonial population was favorably maintained in the pumpless device compared to the conventional method. These results show the advantage of using microfluidic systems for organ culture experiments. Our pumpless device could be applied to a variety of other tissues and organs, and may revolutionize organ culture methods as a whole.


Assuntos
Dispositivos Lab-On-A-Chip , Espermatogênese/fisiologia , Testículo/fisiologia , Animais , Transferência Embrionária/métodos , Desenho de Equipamento , Feminino , Pressão Hidrostática , Masculino , Camundongos , Camundongos Transgênicos , Oócitos , Técnicas de Cultura de Órgãos/instrumentação
12.
Asian J Androl ; 19(3): 350-354, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-26975485

RESUMO

This study investigated the correlation between sperm motion parameters obtained by a computer-assisted semen analyzer and levels of reactive oxygen species in unwashed semen. In total, 847 patients, except for azoospermic patients were investigated. At the time of each patient's first consultation, semen parameters were measured using SMAS™ or CellSoft 3000™, and production of reactive oxygen species was measured using a computer-driven LKB Wallac Luminometer 1251 Analyzer. The patients were divided into two groups: reactive oxygen species - positive and negative. The semen parameters within each group were measured using one of the two computer-assisted semen analyzer systems and then compared. Correlations between reactive oxygen species levels and sperm motion parameters in semen from the reactive oxygen species - positive group were also investigated. Reactive oxygen species were detected in semen samples of 282 cases (33.3%). Sperm concentration (P < 0.01; P < 0.01), motility (P < 0.01; P < 0.05), and progressive motility (P < 0.01; P < 0.01) were markedly lower in the reactive oxygen species - positive group than in the reactive oxygen species - negative group. Among the sperm motion parameters in the reactive oxygen species - positive group, sperm concentration (P < 0.01; P < 0.01), motility (P < 0.05; P < 0.01), mALH (P < 0.05; P < 0.01), and progressive motility (P < 0.05; P < 0.01) also showed inverse correlations with the logarithmic transformed reactive oxygen species levels. Therefore, this study demonstrated that excessive reactive oxygen species in semen damage sperm concentration, motility, and other sperm motion parameters.


Assuntos
Espécies Reativas de Oxigênio/análise , Sêmen/química , Motilidade Espermática/fisiologia , Adulto , Idoso , Diagnóstico por Computador , Humanos , Infertilidade Masculina , Luminescência , Masculino , Pessoa de Meia-Idade , Análise do Sêmen , Adulto Jovem
13.
Surg Endosc ; 31(4): 1999-2005, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27572059

RESUMO

BACKGROUND: Laparoscopic surgical techniques are difficult to learn, and developing such skills involves a steep learning curve. To ensure surgeons achieve a high skill level, it is important to be able to measure and assess their skills. Therefore, it is necessary to understand the performance differences between experienced and novice surgeons, as such information could be used to help surgeons learn laparoscopic skills. We examined the differences in gripping and reaction force between experienced and novice surgeons during laparoscopic surgery. METHODS: We measured the gripping force generated during laparoscopic surgery performed on pigs using forceps with pressure sensors. Several sensors, including strain gauges, accelerometers, and a potentiometer, were attached to the forceps. This study included 4 experienced and 4 novice surgeons. Each subject was asked to elevate the kidney in order to approach the renal hilus using the forceps. Throughout the experiment, we measured the gripping force and reaction force generated during the movement of the forceps in real time. RESULTS: The experienced and novice surgeons exhibited similar reaction force levels. Conversely, gripping force differed significantly between the groups. The experienced and novice surgeons exhibited mean gripping force levels of 3.06 and 7.15 N, respectively. The gripping force standard deviation values for the experienced and novice surgeons were 1.43 and 3.54 N, respectively. The mean and standard deviation gripping force values of the experienced surgeons were significantly lower than those of the novice surgeons (P = 0.015 and P = 0.011, respectively). CONCLUSIONS: This study indicated that experienced surgeons generate weaker but more stable gripping force than novice surgeons during laparoscopic procedures.


Assuntos
Competência Clínica , Força da Mão , Laparoscopia/normas , Instrumentos Cirúrgicos , Urologistas , Animais , Fenômenos Biomecânicos , Internato e Residência , Fenômenos Mecânicos , Sus scrofa , Suínos , Urologia/educação
14.
Am J Cardiol ; 118(9): 1405-1409, 2016 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-27666178

RESUMO

Previous reports have shown that serum elastin fragments (SEFs) may be a useful biomarker for the diagnosis of an acute aortic dissection (AAD). However, because the reference interval of SEFs has not been established, it has not been determined whether SEFs are really useful for the diagnosis of AAD. The purpose of this study was to determine the usefulness of measuring SEFs for the diagnosis of AAD. A total of 42 consecutive patients aged 68 ± 18 years who were diagnosed with an AAD were studied. Patient background and SEF levels were examined on admission. SEF levels were also measured in patients undergoing a medical examination (n = 531, age 54 ± 17 years) to compare with those with an AAD. In the control group, SEF levels increased with age (R = 0.725, p <0.001). Then, we defined the upper limit of the reference interval of SEF levels as the 97.5th percentile of control SEF grouped by decade of life from the sixth to ninth decade. The overall risk of AAD exceeding the upper limit of the reference interval at each decade was 10% (4 of 42). For patients in their 60s and 70s, median SEF levels in the AAD group (89 [77 to 104], 93 [60 to 123] ng/ml, respectively) were not significantly higher than those in the control group (79 [68 to 92], 90 [79 to 106] ng/ml, respectively; p = 0.081 and 0.990, respectively). Our data suggest that measuring SEF levels may not be useful in the diagnosis of an AAD as the upper limit of the reference interval of the SEF level was unexpectedly higher.


Assuntos
Aneurisma Dissecante/diagnóstico , Aneurisma Aórtico/diagnóstico , Elastina/sangue , Doença Aguda , Idoso , Biomarcadores/sangue , Diagnóstico Diferencial , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
15.
Proc Natl Acad Sci U S A ; 113(24): E3394-402, 2016 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-27226296

RESUMO

Invariant natural killer T (iNKT) cells are a subset of innate-like T cells that act as important mediators of immune responses. In particular, iNKT cells have the ability to immediately produce large amounts of IFN-γ upon activation and thus initiate immune responses in various pathological conditions. However, molecular mechanisms that control IFN-γ production in iNKT cells are not fully understood. Here, we report that basic helix-loop-helix transcription factor family, member e40 (Bhlhe40), is an important regulator for IFN-γ production in iNKT cells. Bhlhe40 is highly expressed in stage 3 thymic iNKT cells and iNKT1 subsets, and the level of Bhlhe40 mRNA expression is correlated with Ifng mRNA expression in the resting state. Although Bhlhe40-deficient mice show normal iNKT cell development, Bhlhe40-deficient iNKT cells show significant impairment of IFN-γ production and antitumor effects. Bhlhe40 alone shows no significant effects on Ifng promoter activities but contributes to enhance T-box transcription factor Tbx21 (T-bet)-mediated Ifng promoter activation. Chromatin immunoprecipitation analysis revealed that Bhlhe40 accumulates in the T-box region of the Ifng locus and contributes to histone H3-lysine 9 acetylation of the Ifng locus, which is impaired without T-bet conditions. These results indicate that Bhlhe40 works as a cofactor of T-bet for enhancing IFN-γ production in iNKT cells.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/imunologia , Regulação da Expressão Gênica/imunologia , Proteínas de Homeodomínio/imunologia , Interferon gama/imunologia , Células T Matadoras Naturais/imunologia , Regiões Promotoras Genéticas/imunologia , Proteínas com Domínio T/imunologia , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regulação da Expressão Gênica/genética , Proteínas de Homeodomínio/genética , Imunidade Celular/genética , Interferon gama/genética , Camundongos , Camundongos Knockout , Neoplasias Experimentais/genética , Neoplasias Experimentais/imunologia , Proteínas com Domínio T/genética
16.
Sci Rep ; 6: 21472, 2016 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-26892171

RESUMO

In contrast to cell cultures, particularly to cell lines, tissues or organs removed from the body cannot be maintained for long in any culture conditions. Although it is apparent that in vivo regional homeostasis is facilitated by the microvascular system, mimicking such a system ex vivo is difficult and has not been proved effective. Using the culture system of mouse spermatogenesis, we addressed this issue and devised a simple microfluidic device in which a porous membrane separates a tissue from the flowing medium, conceptually imitating the in vivo relationship between the microvascular flow and surrounding tissue. Testis tissues cultured in this device successfully maintained spermatogenesis for 6 months. The produced sperm were functional to generate healthy offspring with micro-insemination. In addition, the tissue kept producing testosterone and responded to stimulation by luteinizing hormone. These data suggest that the microfluidic device successfully created in vivo-like conditions, in which testis tissue maintained its physiologic functions and homeostasis. The present model of the device, therefore, would provide a valuable foundation of future improvement of culture conditions for various tissues and organs, and revolutionize the organ culture method as a whole.


Assuntos
Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas , Espermatogênese , Espermatozoides/citologia , Testículo/citologia , Testículo/fisiologia , Técnicas de Cultura de Tecidos , Animais , Masculino , Camundongos , Testosterona/biossíntese
17.
Adv Mater ; 27(48): 7951-6, 2015 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-26503073

RESUMO

Crystalline protein assemblies of polyhedra crystal (PhC) can be utilized as solid enzyme containers for long-term storage of enzymes with retention of their enzymatic activity. The enzymes can be released from the crystals at the optimum pH for the enzymatic activity by dissolution of the crystals using in vivo crystal engineering.


Assuntos
Engenharia de Proteínas/métodos , Proteína Quinase C/genética , Proteínas Estruturais Virais/química , Proteínas Estruturais Virais/genética , Animais , Cápsulas , Cristalização , Modelos Moleculares , Estrutura Secundária de Proteína , Células Sf9 , Spodoptera
18.
J Gastroenterol ; 50(11): 1124-33, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25791517

RESUMO

BACKGROUND: Immune tolerance is maintained in the liver, and perturbation of tolerance can lead to immune-mediated liver diseases such as autoimmune hepatitis (AIH). Invariant natural killer T (iNKT) cells and γδT cells have been shown to maintain immune homeostasis as regulatory cells and to play pathogenic roles in immune-mediated diseases as effector cells. We hypothesized that iNKT cells and γδT cells are involved in the maintenance of hepatic immune tolerance and immune-mediated liver disease. METHODS: We measured liver inflammation and the cytokine profiles of liver mononuclear cells in BALB/c wild-type (WT) mice and BALB/c Jα18-deficient (KO) mice lacking iNKT cells. We also examined the role of γδT cells in AIH using liver tissue from AIH patients and control subjects. RESULTS: Spontaneous liver inflammation, hepatocyte damage, and anti-nuclear-antibody production occurred in Jα18 KO mice but not in WT mice. Furthermore, liver mononuclear cells from Jα18 KO mice, but not those from WT mice, produced interleukin-17 (IL-17). γδT cells were the primary producers of the cytokine, and they were more abundant in the livers of Jα18 KO mice than in those of WT mice. In Jα18 KO mice, the administration of anti-γδT-cell-receptor antibody abolished liver inflammation, hepatocyte damage, and IL-17 production. γδT cells accumulated in the livers of AIH patients but not in those of the control subjects. CONCLUSIONS: Our results suggest a protective role for iNKT cells, a pathologic role for γδT cells, and an association between these cells in the pathogenesis of AIH.


Assuntos
Doenças Autoimunes/imunologia , Hepatite/imunologia , Células T Matadoras Naturais/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/análise , Subpopulações de Linfócitos T/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Citocinas/biossíntese , Feminino , Hepatite Animal/imunologia , Humanos , Tolerância Imunológica/imunologia , Interleucina-17/biossíntese , Fígado/imunologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Knockout , Pessoa de Meia-Idade
19.
Adv Urol ; 2015: 495308, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25784932

RESUMO

Objectives. Here we developed a unique training system, a patient specific virtual reality simulator, for laparoscopic renal surgery. To develop the simulator, it was important to first identify the physical properties of the organ. Methods. We recorded the force measured during laparoscopic surgery performed on pigs by using forceps with pressure sensors. Several sensors, including strain gauges, accelerometers, and a potentiometer, are attached to the forceps. Results. Throughout the experiment, we measured the reaction force in response to the forceps movement in real time. Conclusions. The experiment showed the possibility of digitizing these physical properties in humans as well.

20.
Int J Urol ; 22(6): 572-6, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25721949

RESUMO

OBJECTIVES: To describe and to validate a novel patient-specific virtual-reality based simulator for laparoscopic surgery. METHODS: Three surgeons carried out 13 preoperative simulations at Yokohama City University Hospital, Yokohama, Kanagawa, Japan, from 2011 to 2012. The procedures included seven nephrectomies, four partial nephrectomies and two pyeloplasties. We evaluated whether the anatomies reproduced by the simulator matched those encountered during the actual operations. Furthermore, the surgeons were asked to use visual analog scales (from 1 to 5; higher scores are better) to evaluate the anatomical integrity and utility of the simulations, and their intraoperative confidence during the subsequent surgical procedures. RESULTS: The simulator reproduced the patients' anatomies almost perfectly. Only a few minor mistakes were identified. Regarding the surgeons' evaluations of the system, the mean scores for the anatomical integrity and utility of the simulations, and the surgeons' intraoperative confidence were 3.4, 4.2 and 4.1, respectively. In all 13 cases, the surgeons were able to carry out preoperative training with ease, and they stated that the simulator was useful for producing preoperative images. CONCLUSIONS: A patient-specific simulator for laparoscopic renal surgery has been successfully developed. This system correctly reproduces anatomical structures, and it seems to be a useful preoperative training tool.


Assuntos
Rim/anatomia & histologia , Rim/cirurgia , Laparoscopia , Nefrectomia , Modelagem Computacional Específica para o Paciente , Interface Usuário-Computador , Adulto , Idoso , Atitude do Pessoal de Saúde , Feminino , Humanos , Laparoscopia/educação , Masculino , Pessoa de Meia-Idade
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