Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 31
Filtrar
Filtros adicionais











Tipo de estudo
Intervalo de ano
1.
J Nanosci Nanotechnol ; 20(2): 760-768, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31383071

RESUMO

Enhancing the mobility in metal-oxide-semiconductor field-effect transistors (MOSFETs) with narrow channel widths is highly sensitive to the stress effects of Si channel when related advanced strain engineering is introduced and is compatible with semiconductor processes. In practice, layouts have significant effect on the device performance, especially for the protruding gate width on shallow trench isolation structures. The geometric parameter is investigated by systematically analysing an n-channel MOSFET composed of silicon-carbon (SiC) stressors embedded in the source and drain (S/D) regions and a tensile contact etch stop layer (CESL) using three-dimensional finite element simulation. Tensile CESL (1.1 GPa) and a SiC S/D stressor with a carbon mole fraction of 1.65% are loaded on the structure. The difference in the interactive percentages between the mechanical bending effect of the CESL from the top of the poly gate and the downward force of the CESL adjacent to the spacer sidewall of the gate occurs when the protruding gate width is increased. Results indicate that mobility was highly enhanced by approximately 72.5% at a width of approximately 0.2 µm. The mechanical bending effect becomes dominant when the gate width is more than 0.2 µm. Consequently, the mobility gain decays and consequently converges toward a constant.

2.
Environ Toxicol ; 34(8): 921-927, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31066208

RESUMO

Hyperkalemia is often associated with cardiac dysfunction. In this study an earthworm extract (dilong) was prepared from dried Pheretima aspergillum powder and its effect against high-KCl challenge was determined in H9c2 cardiomyoblast cells. H9c2 cells pre-treated with dilong (31.25, 62.5, 125, and 250 mg/mL) for 24 hours, where challenged with different doses of KCl treatment for 3 hours to determine the protective mechanisms of dilong against cardiac fibrosis. High-KCl administration induced mitochondrial injury and elevated the levels of pro-apoptotic proteins. The mediators of fibrosis such as ERK, uPA, SP1, and CTGF were also found to be upregulated in high-KCl condition. However, dilong treatment enhanced IGF1R/PI3k/Akt activation which is associated with cell survival. In addition, dilong also reversed high-KCl induced cardiac fibrosis related events in H9c2 cells and displayed a strong cardio-protective effect. Therefore, dilong is a potential agent to overcome cardiac events associated with high-KCl toxicity.


Assuntos
Mitocôndrias Cardíacas/efeitos dos fármacos , Mioblastos Cardíacos/efeitos dos fármacos , Oligoquetos , Cloreto de Potássio/toxicidade , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular , Sobrevivência Celular , Fibrose , Mioblastos Cardíacos/patologia , Substâncias Protetoras/farmacologia
3.
Clin Nurs Res ; : 1054773819831444, 2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30808207

RESUMO

This study aimed to estimate the effects of a supportive care program on the posttraumatic stress symptoms (PTSSs) of patients with oral cancer after surgery. Participants were divided into two groups. Outcome measurements included the Chinese version of the Davidson trauma scale to examine PTSSs at a clinical follow-up 1 week (T0), 1 month (T1), and 3 months (T2) after hospital discharge. The results indicated that the frequency and severity mean scores of PTSSs for the two groups at T0 were significantly higher than those at T1 and T2. Both the groups and times were significantly different; moreover, the supported group's PTSS score decreased more than that of the nonsupported group. These findings supported the effects of the supportive care program. Health care staff should be aware of the PTSS status of patients with oral cancer who undergo surgery and consider these issues in combination with patients' discharge care plans.

4.
Kidney Blood Press Res ; 43(6): 1822-1831, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30537749

RESUMO

BACKGROUND/AIMS: Hyperglycemia and hyperuricemia are two major disorders of Metabolic syndrome. Kidney plays a crucial role in maintaining the homeostasis of uric acid and glucose. The aim of the study was to examine the changes of renal glucose and uric acid transporters in animals with metabolic syndrome. METHODS: Sprague-Dawley rats were fed with high fructose diet (60%) for 3 months (FR-3) and 5 months (FR-5). At the end study, serum and urine biochemical data were compared. Gene expression and protein abundance of renal GLUT1, GLUT2, GLUT9, SGLT1, SGLT2, UAT and URAT1 was investigated by using RT-PCR and immunohistochemical staining. RESULTS: Metabolic syndrome was induced by high-fructose diet. Systolic blood pressure and proteinuria was significantly increased in FR-5 animals. In kidney tissue, gene expression of GLUT2 and SGLT2 increased significantly in a time dependent manner. GLUT9, SGLT1 and UAT were also significantly upregulated in FR-5. Immunohistochemical study showed a significant increase of SGLT1 in both FR-3 (413.5 ± 88.3% of control, p< 0.001) and FR-5 (677.6 ± 26.5% of control, p< 0.001). Also, SGLT2 protein was increased in both FR-3 (643.1 ± 41.3% of control, p< 0.001) and FR-5 (563.3 ± 21.7% of control, p< 0.001). Fructose rich food also induced increase of UAT by nearly 5-fold in both FR-3 and FR-5 (both p< 0.05) and more than 3-fold of GLUT-9 in FR-3 and FR-5 (both p< 0.05). CONCLUSION: Long term high fructose diet induced metabolic syndrome with increased blood pressure and proteinuria in rats. Metabolic syndrome was associated with dual increase in renal glucose and uric acid transporters, including SGLT1, SGLT2, GLUT2, GLUT9 and UAT.


Assuntos
Frutose/efeitos adversos , Síndrome Metabólica/metabolismo , Transportadores de Ânions Orgânicos/metabolismo , Proteínas de Transporte de Sódio-Glucose/metabolismo , Animais , Epitélio/química , Rim/química , Rim/citologia , Síndrome Metabólica/induzido quimicamente , Ratos , Ratos Sprague-Dawley
5.
Environ Toxicol ; 32(12): 2471-2480, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28856781

RESUMO

Anthocyanins are known cyto-protective agents against various stress conditions. In this study cardio-protective effect of anthocyanins from black rice against diabetic mellitus (DM) was evaluated using a streptozotocin (STZ)-induced DM rat model. Five-week-old male Wistar rats were administered with STZ (55 mg kg-1 , IP) to induce DM; rats in the treatment group received 250 mg oral anthocyanin/kg/day during the 4-week treatment period. DM and the control rats received normal saline through oral gavage. The results reveal that STZ-induced DM elevates myocardial apoptosis and associated proapoptotic proteins but down-regulates the proteins of IGF1R mediated survival signaling mechanism. Furthermore, the functional parameters such as the ejection-fraction and fraction-shortening in the DM rat hearts declined considerably. However, the rats treated with anthocyanins significantly reduced apoptosis and the associated proapoptotic proteins and further increased the survival signals to restore the cardiac functions in DM rats. Anthocyanin supplementation enhances cardiomyocyte survival and restores cardiac function.


Assuntos
Antocianinas/farmacologia , Cardiotônicos/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Coração/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor IGF Tipo 1/metabolismo , Estreptozocina , Animais , Antocianinas/uso terapêutico , Apoptose/efeitos dos fármacos , Cardiotônicos/uso terapêutico , Sobrevivência Celular/efeitos dos fármacos , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/fisiopatologia , Coração/fisiopatologia , Masculino , Miocárdio/patologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/patologia , Ratos Wistar , Transdução de Sinais
6.
Nephron ; 137(2): 91-98, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28637038

RESUMO

BACKGROUND: Both cardiovascular calcification and autonomic dysfunction are frequently encountered in hemodialysis patients. We aimed to investigate the relationship between cardiovascular calcification and heart rate variability (HRV) and their influence on long-term outcome. METHODS: Seventy-eight hemodialysis patients underwent echocardiogram and radiography of the pelvis and hands to identify valvular and vascular calcification. HRV was evaluated using a commercial machine. RESULTS: Based on the average, the patients were divided into higher and lower subgroups of high frequency (HF) and low frequency (LF) respectively. Patients with higher LF were younger and were found to have a lower proportion of diabetes. Their hemoglobin, albumin, and bone morphogenic protein (BMP)-7 levels were significantly higher and both high-sensitive C-reactive protein (hs-CRP) and osteoprotegerin levels were lower (all p < 0.05). In patients of the higher HF group, the proportion of diabetes was lower but they were found to have higher levels of BMP-7 and lower levels of hs-CRP, interleukin-6 (all p < 0.05). Significantly higher LF and HF were noted in patients without vascular calcification, but only hand artery (HA) calcification was negatively correlated with both LF and HF in multivariate analysis. Low LF and high hs-CRP were the independent predictors of mortality. Coexistence of low LF band and HA calcification was associated with the worse outcome. CONCLUSIONS: Abnormal autonomic nervous function was closely related to inflammation and mortality in hemodialysis patients. Calcification of HA was associated with autonomic dysfunction and patients with lower autonomic tone and HA calcification had the highest mortality rate in this population.


Assuntos
Doenças do Sistema Nervoso Autônomo/complicações , Doenças do Sistema Nervoso Autônomo/mortalidade , Nefropatias/complicações , Nefropatias/mortalidade , Diálise Renal , Calcificação Vascular/complicações , Calcificação Vascular/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Doenças do Sistema Nervoso Autônomo/terapia , Proteína Morfogenética Óssea 7/sangue , Proteína C-Reativa/análise , Estudos de Coortes , Nefropatias Diabéticas/complicações , Nefropatias Diabéticas/mortalidade , Eletrocardiografia , Feminino , Frequência Cardíaca , Humanos , Interleucina-6/sangue , Nefropatias/terapia , Masculino , Pessoa de Meia-Idade , Osteoprotegerina/sangue , Pelve/diagnóstico por imagem , Valor Preditivo dos Testes , Estudos Prospectivos , Calcificação Vascular/terapia
7.
Int J Mol Sci ; 17(9)2016 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-27657062

RESUMO

Doxorubicin (Dox) is extensively used for chemotherapy in different types of cancer, but its use is limited to because of its cardiotoxicity. Our previous studies found that doxorubicin-induced insulin-like growth factor II receptor (IGF-IIR) accumulation causes cardiomyocytes apoptosis via down-regulation of HSF1 pathway. In these studies, we demonstrated a new mechanism through which anthocyanin protects cardiomyoblast cells against doxorubicin-induced injury. We found that anthocyanin decreased IGF-IIR expression via estrogen receptors and stabilized heat shock factor 1 (HSF1) to inhibit caspase 3 activation and apoptosis of cardiomyocytes. Therefore, the phytoestrogen from plants has been considered as another potential treatment for heart failure. It has been reported that the natural compound anthocyanin (ACN) has the ability to reduce the risk of cardiovascular disease (CVD). Here, we demonstrated that anthocyanin acts as a cardioprotective drug against doxorubicin-induced heart failure by attenuating cardiac apoptosis via estrogen receptors to stabilize HSF1 expression and down-regulated IGF-IIR-induced cardiomyocyte apoptosis.

8.
Medicine (Baltimore) ; 95(11): e3032, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26986120

RESUMO

Stroke is one of the most common causes of physical disability, and early, intensive, and repetitive rehabilitation exercises are crucial to the recovery of stroke survivors. Unfortunately, research shows that only one third of stroke patients actually perform recommended exercises at home, because of the repetitive and mundane nature of conventional rehabilitation exercises. Thus, to motivate stroke survivors to engage in monotonous rehabilitation is a significant issue in the therapy process. Game-based rehabilitation systems have the potential to encourage patients continuing rehabilitation exercises at home. However, these systems are still rarely adopted at patients' places. Discovering and eliminating the obstacles in promoting game-based rehabilitation at home is therefore essential. For this purpose, we conducted a study to collect and analyze the opinions and expectations of stroke patients and clinical therapists. The study is composed of 2 parts: Rehab-preference survey - interviews to both patients and therapists to understand the current practices, challenges, and expectations on game-based rehabilitation systems; and Rehab-compatibility survey - a gaming experiment with therapists to elaborate what commercial games are compatible with rehabilitation. The study is conducted with 30 outpatients with stroke and 19 occupational therapists from 2 rehabilitation centers in Taiwan. Our surveys show that game-based rehabilitation systems can turn the rehabilitation exercises more appealing and provide personalized motivation for various stroke patients. Patients prefer to perform rehabilitation exercises with more diverse and fun games, and need cost-effective rehabilitation systems, which are often built on commodity hardware. Our study also sheds light on incorporating the existing design-for-fun games into rehabilitation system. We envision the results are helpful in developing a platform which enables rehab-compatible (i.e., existing, appropriately selected) games to be operated on commodity hardware and brings cost-effective rehabilitation systems to more and more patients' home for long-term recovery.


Assuntos
Reabilitação do Acidente Vascular Cerebral , Jogos de Vídeo/psicologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Ocupacional/psicologia , Preferência do Paciente , Adulto Jovem
9.
Materials (Basel) ; 9(9)2016 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-28773838

RESUMO

Stress-induced failure is a critical concern that influences the mechanical reliability of an indium tin oxide (ITO) film deposited on a transparently flexible polyethylene terephthalate (PET) substrate. In this study, a cycling bending mechanism was proposed and used to experimentally investigate the influences of compressive and tensile stresses on the mechanical stability of an ITO film deposited on PET substrates. The sheet resistance of the ITO film, optical transmittance of the ITO-coated PET substrates, and failure scheme within the ITO film were measured to evaluate the mechanical stability of the concerned thin films. The results indicated that compressive and tensile stresses generated distinct failure schemes within an ITO film and both led to increased sheet resistance and optical transmittance. In addition, tensile stress increased the sheet resistance of an ITO film more easily than compressive stress did. However, the influences of both compressive and tensile stress on increased optical transmittance were demonstrated to be highly similar. Increasing the thickness of a PET substrate resulted in increased sheet resistance and optical transmittance regardless of the presence of compressive or tensile stress. Moreover, J-Integral, a method based on strain energy, was used to estimate the interfacial adhesion strength of the ITO-PET film through the simulation approach enabled by a finite element analysis.

10.
Materials (Basel) ; 9(10)2016 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-28773969

RESUMO

The suitability of stacked thin films for next-generation display technology was analyzed based on their properties and geometrical designs to evaluate the mechanical reliability of transparent conducting thin films utilized in flexural displays. In general, the high bending stress induced by various operation conditions is a major concern regarding the mechanical reliability of indium-tin-oxide (ITO) films deposited on polyethylene terephthalate (PET) substrates; mechanical reliability is commonly used to estimate the flexibility of displays. However, the pattern effect is rarely investigated to estimate the mechanical reliability of ITO/PET films. Thus, this study examined the flexible content of patterned ITO/PET films with two different line widths by conducting bending tests and sheet resistance measurements. Moreover, a stress-strain simulation enabled by finite element analysis was performed on the patterned ITO/PET to explore the stress impact of stacked film structures under various levels of flexural load. Results show that the design of the ITO/PET film can be applied in developing mechanically reliable flexible electronics.

11.
Taiwan J Obstet Gynecol ; 54(6): 641-6, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26700978

RESUMO

OBJECTIVE: In 1991, researchers reported that a modest preovulatory increase in serum progesterone levels is associated with lower pregnancy rates and higher incidence of pregnancy loss in in vitro fertilization (IVF). We wonder whether embryo transfer (ET) in assisted reproductive technology (ART) cycles in patients with premature progesterone rise (PPR) have a negative impact on the clinical pregnancy rates (CPRs) and/or live birth rates (LBRs) in our series. Consequently, will blastocyst transfer reverse the negative impact? MATERIALS AND METHODS: This noninterventional, retrospective, observational tertiary center study was conducted between January 2010 and December 2012. All fresh ET cycles with serum progesterone levels measured (n = 599) on the day of hCG administration were analyzed. RESULTS: Sera lutenizing hormone (LH), E2, and progesterone (P) were measured and analyzed. The CPRs of cycles in patients with p ≤ 1.5 ng/mL (low) versus those with p > 1.5 ng/mL (high) were 37.04% versus 41.03% [odds ratio (OR) = 1.18, 95% confidence interval (CI): 0.728-1.920; p = 0.50). The LBRs of cycles in patients with low progesterone level versus those with PPR were 30.52% versus 34.62% (OR = 1.21, 95% CI: 0.729-1.992; p = 0.47). No statistically significant association was detected. We further analyzed the outcomes according to different stages of ET and found that blastocyst (D5) ET significantly increase the LBRs as compared with cleavage stage (D2/D3) ET in the PPR group (44.44% versus 21.43%; p = 0.043). CONCLUSION: PPR did not significantly compromise the clinical outcomes in this series. However, shifting to blastocyst transfer probably could increase the live birth in cycles with PPR.


Assuntos
Blastocisto/citologia , Transferência Embrionária , Progesterona/sangue , Técnicas de Reprodução Assistida , Adulto , Técnicas de Cultura Embrionária , Feminino , Humanos , Nascimento Vivo , Hormônio Luteinizante/sangue , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
12.
Materials (Basel) ; 8(8): 5121-5137, 2015 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-28793495

RESUMO

three-dimensional integrated circuit (3D-IC) structure with a significant scale mismatch causes difficulty in analytic model construction. This paper proposes a simulation technique to introduce an equivalent material composed of microbumps and their surrounding wafer level underfill (WLUF). The mechanical properties of this equivalent material, including Young's modulus (E), Poisson's ratio, shear modulus, and coefficient of thermal expansion (CTE), are directly obtained by applying either a tensile load or a constant displacement, and by increasing the temperature during simulations, respectively. Analytic results indicate that at least eight microbumps at the outermost region of the chip stacking structure need to be considered as an accurate stress/strain contour in the concerned region. In addition, a factorial experimental design with analysis of variance is proposed to optimize chip stacking structure reliability with four factors: chip thickness, substrate thickness, CTE, and E-value. Analytic results show that the most significant factor is CTE of WLUF. This factor affects microbump reliability and structural warpage under a temperature cycling load and high-temperature bonding process. WLUF with low CTE and high E-value are recommended to enhance the assembly reliability of the 3D-IC architecture.

13.
Biochem Biophys Res Commun ; 438(2): 359-63, 2013 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-23896603

RESUMO

Lead intoxication is usually insidious and may cause a variety of complications such as kidney damage and hypertension. The role of intrarenal renin-angiotensin system (RAS) in lead-induced nephropathy has not been investigated. Adult male Sprague-Dawley rats were fed with water containing 250ppm of lead acetate (lead group) and deionized water (control group) for 4weeks. Another two groups started to receive intraperitoneal captopril (50mg/kg/d) or losartan (10mg/kg/d) after 2weeks of lead feeding and continued for another 2weeks. Immunoblotting was used to analyze the protein amount of intrarenal RAS components and transforming growth factor-beta (TGF-ß). Compared with control group, lead exposure resulted in increased proteinuria after 2-week treatment (4.2±0.9mg/100g vs. 1.8±0.8mg/100g, p<0.05) and 4-week (5.2±1.7mg/100g, p<0.05). Serum creatinine level was increased (0.40±0.2 vs. 0.3 ±.04mg/dL, p<0.05) and calculated glomerular filtration rate (GFR) was decreased (2.68±1.03 vs. 3.37±0.11mL/min, p<0.05). Intrarenal angiotensin converting enzyme (ACE), angiotensin II (ANG II), angiotensin II type 1 receptor (AT1R) and transforming growth factor-beta (TGF-ß) were upregulated in lead group. Captopril and losartan administration reduced proteinuria significantly (3.0±0.50mg/100g of captopril and 2.7±0.4mg/100g of losartan group) and lowered systolic blood pressure when compared with lead group. Furthermore, serum creatinine levels and GFR were improved by RAS blockade. Captopril treatment significantly reduced protein abundance of ACE, ANG II, AT1R and TGF-ß. Losartan treatment also decreased ANG II and TGF-ß. We concluded that lead exposure elicited intrarenal RAS activation with associated proteinuria and impaired renal function. RAS blockade was effective in alleviating lead-associated kidney injury and lowering blood pressure.


Assuntos
Regulação da Expressão Gênica , Nefropatias/induzido quimicamente , Intoxicação por Chumbo/fisiopatologia , Sistema Renina-Angiotensina/fisiologia , Angiotensina II/metabolismo , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Pressão Sanguínea , Captopril/farmacologia , Taxa de Filtração Glomerular , Losartan/farmacologia , Masculino , Peptidil Dipeptidase A/metabolismo , Proteinúria/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina/metabolismo , Sistema Renina-Angiotensina/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo
14.
Ther Apher Dial ; 16(6): 573-9, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23190518

RESUMO

High prevalence of depression has been reported in patients with end stage kidney disease and depression is associated with increased morbidity and mortality. We aimed to investigate the prevalence of depression in patients receiving standard hemodialysis (SHD) and hemodiafiltration (HDF) and compare the associated factors between these treatment modalities. The Beck Depression Inventory (BDI) was used to survey for major depressive symptoms. Demographic and biochemical data were reviewed and collected. Point prevalence of depression in HDF patients was significantly lower than SHD patients (23.9% vs. 43.1%, P < 0.05). The BDI score was also higher in SHD than HDF group (13.2 ± 11.6 vs. 8.7 ± 11.2, P < 0.05). SHD patients with major depressive symptoms had significantly lower levels of hemoglobin, albumin, creatinine, sodium and hand grip strength but had higher prevalence of diabetes and high sensitivity C-reactive protein (hs-CRP) levels. In HDF patients, phosphorus level was significantly lower in patients with major depressive symptoms. Logistic regression analysis revealed that hs-CRP, serum sodium and hand grip strength were significantly associated with major depressive symptoms in patients treated with SHD; while serum phosphorus was identified in HDF groups. We concluded that prevalence of depression was high in dialysis patients. Patients receiving HDF had a lower mean BDI score and a nearly 50% lower prevalence rate of major depressive symptoms than that of SHD. Factors associated with depression were different between two modalities.


Assuntos
Transtorno Depressivo Maior/epidemiologia , Hemodiafiltração , Falência Renal Crônica/terapia , Diálise Renal , Idoso , Proteína C-Reativa/metabolismo , Transtorno Depressivo Maior/fisiopatologia , Feminino , Força da Mão , Hemoglobinas/metabolismo , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fósforo/sangue , Prevalência , Escalas de Graduação Psiquiátrica , Sódio/sangue , Uremia/epidemiologia
15.
J Biomed Biotechnol ; 2009: 104716, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-20168979

RESUMO

MicroRNAs (miRNAs) downregulate gene expression by binding to the partially complementary sites in the 3' untranslated region (UTR) of target mRNAs. Several methods, such as Northern blot analysis, quantitative real-time RT-PCR, microarray, and the luciferase reporter system, are commonly used to quantify the relative level or activity of miRNAs. The disadvantage of these methods is the requirement for cell lysis, which means that several sets of wells/dishes of cells must be prepared to monitor changes in miRNA activity in time-course studies. In this study, we developed a multisampling reporter system in which two secretable bioluminescence-generating enzymes are employed, one as a reporter and the other as an internal control. The reporters consist of a pair of vectors containing the Metridia luciferase gene, one with and one without a duplicated miRNA targeting sequence at their 3'UTR, while the other vector coding for the secreted alkaline phosphatase gene is used as an internal control. This method allows miRNA activity to be monitored within the same population of cells over time by withdrawing aliquots of the culture medium. The practicability and benefits of this system are addressed in this report.


Assuntos
Genes Reporter , Técnicas Genéticas , MicroRNAs/análise , MicroRNAs/genética , Catequina/análogos & derivados , Catequina/farmacologia , Células Hep G2 , Humanos , MicroRNAs/metabolismo
16.
Am J Physiol Renal Physiol ; 295(4): F923-31, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18632795

RESUMO

The impact of acute anal stretch on the pelvic-urethra reflex potentiation was examined in urethane-anesthetized rats by recording the external urethra sphincter electromyogram activity evoked by the pelvic afferent stimulation. Test stimulation (1 stimulation/30 s) evoked a baseline reflex activity with a single action potential that was abolished by gallamine (5 mg/kg iv). On the other hand, the repetitive stimulation (1 stimulation/1 s) induced spinal reflex potentiation (SRP) that was attenuated by intrathecal 6-cyano-7-nitroquinoxaline-2,4-dione (a glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionat receptor antagonist, 100 microM, 10 microl) and d-2-amino-5-phosphonovalerate [a glutamatergic N-methyl-D-aspartate (NMDA) antagonist, 100 microM, 10 microl]. Acute anal stretch using a mosquito clamp with a distance of 4 mm exhibited no effect, whereas distances of 8 mm attenuated and 12 mm abolished the repetitive stimulation-induced SRP. Intrathecal NMDA (100 microM, 10 microl) reversed the abolition on SRP caused by anal stretch. On the other hand, pretreated bicuculline [gamma-aminobutyric acid (GABA) A receptor antagonist, 100 microM, 10 microl] but not hydroxysaclofen (GABAB receptor antagonist) counteracted the abolition on the repetitive stimulation-induced SRP caused by the anal stretch. All of the results suggested that anal stretch may be used as an adjunct to assist voiding dysfunction in patients with overactive urethra sphincter and that GABAergic neurotransmission is important in the neural mechanisms underlying external urethra sphincter activity inhibited by anal stretch.


Assuntos
Canal Anal/inervação , Inibição Neural/fisiologia , Reflexo/fisiologia , Uretra/inervação , Micção/fisiologia , Ácido gama-Aminobutírico/fisiologia , 2-Amino-5-fosfonovalerato/farmacologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Canal Anal/fisiologia , Anestesia , Animais , Bicuculina/farmacologia , Estimulação Elétrica , Eletromiografia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Antagonistas GABAérgicos/farmacologia , Trietiodeto de Galamina/farmacologia , N-Metilaspartato/agonistas , N-Metilaspartato/antagonistas & inibidores , N-Metilaspartato/metabolismo , Fármacos Neuromusculares não Despolarizantes/farmacologia , Pelve/inervação , Ratos , Ratos Wistar , Reflexo/efeitos dos fármacos , Medula Espinal/fisiologia , Uretra/fisiologia
17.
Am J Physiol Renal Physiol ; 295(5): F1324-35, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18632800

RESUMO

Spinal cord-mediated cross-organ sensitization between the uterus and the lower urinary tract may underlie the high concurrence of obstetrical/gynecological inflammation and chronic pelvic pain syndrome characterized by urogenital pain. However, the neural pathway and the neurotransmitters involved are still unknown. We tested the hypothesis that the excitation of capsaicin-sensitive primary afferent fibers arising from the uterus through the stimulation of transient receptor potential vanilloid 1 (TRPV1) induces cross-organ sensitization on the pelvic-urethra reflex activity. Capsaicin (1-1,000 microM, 0.05 ml) was instilled into the uterus to induce cross-organ reflex sensitization. Activation of capsaicin-sensitive primary afferent fibers by capsaicin instillation into the uterine horn sensitized the pelvic-urethra reflex activity that was reversed by an intrauterine pretreatment with capsaizepine, a TRPV1-selective antagonist. Intrathecal injection of AP5, a glutamatergic N-methyl-D-aspartate (NMDA) antagonist, and Co-101244, an NMDA NR2B-selective antagonist, both abolished the cross-organ reflex sensitization caused by capsaicin instillation. These results demonstrated that TRPV1 plays a crucial role in contributing to the capsaicin-sensitive primary afferent fibers mediating the glutamatergic NMDA-dependent cross-organ sensitization between the uterus and the lower urinary tract when there is a tissue injury.


Assuntos
Capsaicina/farmacologia , Receptores de N-Metil-D-Aspartato/metabolismo , Reflexo/fisiologia , Canais de Cátion TRPV/fisiologia , Útero/fisiologia , 2-Amino-5-fosfonovalerato/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Anestesia , Animais , Capsaicina/análogos & derivados , Relação Dose-Resposta a Droga , Estimulação Elétrica , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Músculo Liso/efeitos dos fármacos , Músculo Liso/inervação , Músculo Liso/metabolismo , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/fisiologia , Dor Pélvica/fisiopatologia , Fosforilação/efeitos dos fármacos , Piperidinas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Reflexo/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Canais de Cátion TRPV/antagonistas & inibidores , Útero/efeitos dos fármacos , Útero/inervação
18.
Am J Physiol Endocrinol Metab ; 295(3): E559-68, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18577691

RESUMO

Cross-organ sensitization between the uterus and the lower urinary tract (LUT) underlies the high concurrence of pelvic pain syndrome and LUT dysfunctions, and yet the role of gonadal steroids is still unknown. We tested the hypothesis that cross-organ sensitization on pelvic-urethra reflex activity caused by uterine capsaicin instillation is estrous cycle dependent. When compared with the baseline reflex activity (1.00 +/- 0.00 spikes/stimulation), uterine capsaicin instillation significantly increased reflex activity (45.42 +/- 9.13 spikes/stimulation, P < 0.01, n = 7) that was corroborated by an increase in phosphorylated NMDA NR2B (P < 0.05, n = 4) but not NR2A subunit (P > 0.05, n = 4) expression. Both intrauterine pretreatment with capsazepine (5.02 +/- 2.11 spikes/stimulation, P < 0.01, n = 7) and an intrathecal injection of AP5 (3.21 +/- 0.83 spikes/stimulation, P < 0.01, n = 7) abolished the capsaicin-induced cross-organ sensitization and the increment in the phosphorylated NR2B level (P < 0.05, n = 4). The degrees of the cross-organ sensitization increased in a dose-dependent manner with the concentration of instilled capsaicin from 100 to 300 microM in both the proestrus and metestrus stages, whereas they weakened when the concentrations were higher than 1,000 microM. Moreover, the cross-organ sensitization caused by the uterine capsaicin instillation increased significantly in the rats during the proestrus stage when compared with the metestrus stage (P < 0.01, n = 7). These results suggest that estrogen levels might modulate the cross-organ sensitization between the uterus and the urethra and underlie the high concurrence of pelvic pain syndrome and LUT dysfunctions.


Assuntos
Ciclo Estral/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologia , Reflexo/fisiologia , Canais de Cátion TRPV/fisiologia , Uretra/fisiologia , Útero/fisiologia , Animais , Western Blotting , Capsaicina/farmacologia , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Injeções Espinhais , N-Metilaspartato/fisiologia , Dor Pélvica/fisiopatologia , Fosforilação , Pressão , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/agonistas , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Canais de Cátion TRPV/efeitos dos fármacos , Uretra/efeitos dos fármacos , Doenças Urológicas/fisiopatologia , Útero/efeitos dos fármacos
19.
Am J Physiol Renal Physiol ; 293(3): F790-800, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17376759

RESUMO

The purposes of this study were to investigate whether the pelvic nerve-to-external urethra sphincter (EUS) reflex potentiation can be induced under physiological conditions and to determine whether glutamatergic neurotransmission is involved in the reflex potentiation. Stimulation-evoked reflex activities, during rhythmic bladder contractions caused by a continuous saline infusion, in 21 anesthetized rats were recorded with/without the intrathecal administration of 10 microl of CNQX (a glutamatergic AMPA receptor antagonist; 100 microM) and APV( a glutamatergic NMDA receptor antagonist; 100 microM). Reflex activities became potentiated following the increment of intravesical pressure (IVP) during the storage phase (2.39 +/- 0.28 spikes/mmHg, n = 21) and the ascending period of the voiding phase (1.46 +/- 0.35 spikes/mmHg, n = 21) and decreased following the decrement of IVP during the descending period of the voiding phase (1.50 +/- 0.33 spikes/mmHg, n = 21). Although it is characterized by a low IVP, a postvoiding reflex potentiation in stimulation-evoked activities was elicited at the critical period after a voiding contraction had just finished (23.95 +/- 8.96 spikes/mmHg, n = 21). The slope of the regression line of evoked activities vs. the IVP during the storage phase was significantly (P < 0.01) higher than that of the ascending and descending periods of the voiding phase, but there was no statistical difference between the ascending and the descending periods (P > 0.05). In addition, the slope of the regression line of posttetanic reflex potentiation was significantly higher than that of the storage phase (P < 0.01). All the slopes of the regression lines decreased after intrathecal CNQX administration (from 3.15 +/- 0.44, 2.10 +/- 0.57, 2.13 +/- 0.53, and 21.30 +/- 3.41 to 0.83 +/- 0.31, 0.74 +/- 0.12, 0.76 +/- 0.12, and 4.31 +/- 3.71 spikes/mmHg in storage, ascending and descending period of the voiding phase, and postvoiding potentiation, respectively; all P < 0.01, n = 10). The slopes of the regression lines became almost horizontal after intrathecal APV administration (from 3.15 +/- 0.44, 2.10 +/- 0.57, 2.13 +/- 0.53, and 21.30 +/- 3.41 to 0.16 +/- 0.12, 0.21 +/- 0.07, 0.18 +/- 0.05, and 0.23 +/- 0.76 spikes/mmHg in storage, ascending and descending period of voiding phase, and postvoiding potentiation, respectively; all P < 0.01, n = 10). Our results suggest that a potentiation in the pelvic nerve-to-EUS reflex can be induced under physiological conditions and the glutamatergic mechanism appears to be involved in this reflex potentiation.


Assuntos
N-Metilaspartato/metabolismo , Pelve/inervação , Nervos Periféricos/fisiologia , Reflexo/fisiologia , Uretra/inervação , Potenciais de Ação , Anestesia , Animais , Eletromiografia , Eletrofisiologia , Feminino , Ratos , Ratos Wistar , Medula Espinal/fisiologia
20.
Am J Physiol Renal Physiol ; 292(6): F1791-801, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17287199

RESUMO

The current study investigates whether the spinal pelvic nerve-to-external urethra sphincter (EUS) reflex potentiation can be induced by a mechanical stimulation and whether the glutamatergic mechanism is involved in yielding such a reflex potentiation. The external urethra sphincter electromyogram (EUSE) activity, evoked by a single or by repetitive pelvic nerve stimulation, in 30 anesthetized rats was recorded with/without bladder saline distension. Without saline distension (0 cmH(2)O), a single pulse nerve stimulation evoked a single action potential in the reflex activity, whereas repetitive pelvic stimulation and saline distension (6 approximately 20 cmH(2)O) both elicited a long-lasting reflex potentiation (20.05 +/- 3.21 and 75.01 +/- 9.87 spikes/stimulation, respectively). The saline distension-induced pelvic nerve-to-EUS reflex potentiation was abolished by D-2-amino-5-phosphonovalerate [APV; a glutamatergic N -methyl-D-aspartic acid (NMDA) receptor antagonist; 100 microM, 10 microl, 1.72 +/- 0.31 spikes/stimulation] and attenuated by 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxaline [NBQX; a glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptor antagonist; 100 microM, 10 microl, 26.16 +/- 7.27 spikes/stimulation], but was not affected by bicuculline (a GABAergic antagonist; 100 microM, 10 microl, 53.62 +/- 15.54 spikes/stimulation). Intrathecal administration of glutamate (31.12 +/- 8.25 spikes/stimulation, 100 microM, 10 microl) and NMDA (26.25 +/- 4.12 spikes/stimulation, 100 microM, 10 microl) both induced a long-lasting pelvic nerve-to-EUS reflex potentiation without saline distension, which was similar to the findings observed from saline distension only. The duration of the contraction wave of the urethra was elongated by the saline distension-induced pelvic nerve-to-EUS reflex potentiation, whereas the peak pressure of the contraction wave was not affected. Our findings suggest that saline distension in the bladder elicits a pelvic nerve-to-EUS reflex potentiation and the glutamatergic mechanism contributes to the presence of such a reflex potentiation.


Assuntos
Ácido Glutâmico/fisiologia , Pelve/inervação , Nervos Periféricos/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologia , Reflexo/fisiologia , Uretra/inervação , Potenciais de Ação/fisiologia , Anestesia , Animais , Dilatação , Eletromiografia , Feminino , Agonistas GABAérgicos/farmacologia , Antagonistas GABAérgicos/farmacologia , Injeções Espinhais , Músculo Esquelético/fisiologia , Estimulação Física , Pressão , Ratos , Ratos Wistar , Receptores de AMPA/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Traumatismos da Medula Espinal/fisiopatologia , Uretra/fisiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA