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Argonaute (AGO), a component of RNA-induced silencing complexes (RISCs), is a representative RNA-binding protein (RBP) known to bind with mature microRNA (miRNA) and is directly involved in post-transcriptional gene silencing. However, despite the biological significance of miRNA, the roles of other micro RNA-binding proteins (miRBPs) remain unclear in regulation of miRNA loading, dissociation from RISC, and extracellular release. In this study, we perform protein arrays to profile miRBPs and identify 118 RNA-binding proteins directly binding with miRNAs. Among those proteins, RBP quaking (QKI) inhibits extracellular release of mature microRNA let-7b by controlling the loading of let-7b into extracellular vesicles via additional miRBPs such as hnRNPD/AUF1 and hnRNPK. The enhanced extracellular release of let-7b after QKI depletion activates the Toll-like Receptor 7 (TLR7) and promotes the production of proinflammatory cytokines in recipient cells, leading to brain inflammation in mouse cortex. Thus, this study reveals contribution of QKI to the inhibition of brain inflammation via regulation of extracellular let-7b release.
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Immune checkpoint inhibitors (ICIs) are widely used in cancer immunotherapy, requiring effective methods for response monitoring. This study evaluated changes in 18F-2-fluoro-2-deoxy-D-glucose (FDG) and 18F-fluorothymidine (FLT) uptake by tumors following ICI treatment as potential imaging biomarkers in mice. Tumor uptakes of 18F-FDG and 18F-FLT were measured and compared between the ICI treatment and control groups. A combined imaging index of glucose-thymidine uptake ratio (GTR) was defined and compared between groups. In the ICI treatment group, tumor growth was effectively inhibited, and higher proportions of immune cells were observed. In the early phase, 18F-FDG uptake was higher in the treatment group, whereas 18F-FLT uptake was not different. There was no difference in 18F-FDG uptake between the two groups in the late phase. However, 18F-FLT uptake of the control group was markedly increased compared with the ICI treatment group. GTR was consistently higher in the ICI treatment group in the early and late phases. After ICI treatment, changes in tumor cell proliferation were observed with 18F-FLT, whereas 18F-FDG showed altered metabolism in both tumor and immune cells. A combination of 18F-FLT and 18F-FDG PET, such as GTR, is expected to serve as a potentially effective imaging biomarker for monitoring ICI treatment.
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Fluordesoxiglucose F18 , Neoplasias , Animais , Biomarcadores , Didesoxinucleosídeos , Fluordesoxiglucose F18/uso terapêutico , Glucose/uso terapêutico , Inibidores de Checkpoint Imunológico/farmacologia , Inibidores de Checkpoint Imunológico/uso terapêutico , Camundongos , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/uso terapêutico , Timidina/farmacologiaRESUMO
A total of 150 growing pigs ([Landrace × Yorkshire] × Duroc) with an initial average body weight (BW) of 24.45 kg were used in a 6-week trial to estimate the optimum lysine to glutamic acid ratio in pigs fed low-protein diets supplemented with increasing level of synthetic glutamic acid (Glu). Pigs were randomly allotted to 5 dietary treatments consisting of either control diet (CON) formulated to have 157 g crude protein (CP) or negative control diets (NC, NC1, NC2 and NC3) with 20 g CP reduction and addition of Glu (1.1, 3.9, 6.8 and 9.6 g/kg feed respectively). Supplementing the increasing level of Glu to low CP diets did not exert any linear or quadratic responses in the growth performance parameters as well as nutrient digestibility. The serum creatinine concentration in pigs receiving CON diet showed trends (p = 0.063) in increment compared with pigs receiving NC diet. However, with the increase in the supplementation of Glu, there were no linear or quadratic responses on serum blood urea nitrogen (BUN) and creatinine concentrations. There was a tendency in the reduction (p = 0.088, p = 0.064) of backfat thickness and lean percentage, respectively, at week 3 and a trend in the reduction (p = 0.092) in lean percentage at week 6 in pigs fed NC diet compared with those fed CON diet. The increase in the supplemental level of Glu tended to show quadratic responses in the backfat thickness and lean percentage at week 3 and 6. In conclusion, the growth performance parameters as well as carcass traits with Lys: Glu ratio 1: 2.71 were very close with the mean values of CON diet indicating that 6.8 g Glu when supplemented to 2% CP reduced diet could achieve the comparable growth performance and carcass trait as that of standard basal diet.
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Ração Animal , Ácido Glutâmico , Ração Animal/análise , Animais , Composição Corporal , Peso Corporal , Dieta/veterinária , Dieta com Restrição de Proteínas/veterinária , Suplementos Nutricionais , Ácido Glutâmico/farmacologia , Lisina/farmacologia , SuínosRESUMO
There has been considerable interest in the clinical use of exosomes as delivery vehicles for treatments as well as for promising diagnostic biomarkers, but the physiological distribution of exosomes must be further elucidated to validate their efficacy and safety. Here, we aimed to develop novel methods to monitor exosome biodistribution in vivo using positron emission tomography (PET) and optical imaging. Exosomes were isolated from cultured mouse breast cancer cells and labeled for PET and optical imaging. In mice, radiolabeled and fluorescently labeled exosomes were injected both via lymphatic and hematogenous metastatic routes. PET and fluorescence images were obtained and quantified. Radioactivity and fluorescence intensity of ex vivo organs were measured. PET signals from exosomes in the lymphatic metastatic route were observed in the draining sentinel lymph nodes. Immunohistochemistry revealed greater exosome uptake in brachial and axillary versus inguinal lymph nodes. Following administration through the hematogenous metastasis pathway, accumulation of exosomes was clearly observed in the lungs, liver, and spleen. Exosomes from tumor cells were successfully labeled with 64Cu (or 68Ga) and fluorescence and were visualized via PET and optical imaging, suggesting that this simultaneous and rapid labeling method could provide valuable information for further exosome translational research and clinical applications.
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Exossomos , Corantes Fluorescentes/farmacocinética , Imagem Multimodal/métodos , Animais , Carbocianinas/química , Carbocianinas/farmacocinética , Radioisótopos de Cobre , Vias de Administração de Medicamentos , Exossomos/química , Feminino , Corantes Fluorescentes/administração & dosagem , Corantes Fluorescentes/química , Radioisótopos de Gálio , Compostos Heterocíclicos com 1 Anel/química , Injeções Intravenosas , Marcação por Isótopo/métodos , Camundongos Endogâmicos BALB C , Tomografia por Emissão de Pósitrons/métodos , Distribuição TecidualRESUMO
B-RafV600E oncogene mutation occurs in various cancers and is associated with tumor initiation. However, genetic modification of B-RafV600E in cells induces MAPK activation and results in oncogene-induced senescence. Overcoming the oncogene-induced senescence by B-RafV600E requires activation of another oncogene pathway, such as AKT signaling. In the present study, we explored the factors involved in overcoming the senescence program in cells activated by B-RafV600E and AKT signaling. B-RafV600E activation caused a feedback inhibition of AKT phosphorylation and resulted in downregulation of FoxM1, one of the AKT downstream components. AKT activation by PTEN downregulation induced FoxM1 expression, and co-expression of B-RafV600E and FoxM1 overcame the cellular senescence. These observations suggested that FoxM1 is critical downstream gene of AKT and functions to overcome B-RafV600E-induced senescence.
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Transformação Celular Neoplásica/genética , Senescência Celular/genética , Fibroblastos/metabolismo , Proteína Forkhead Box M1/genética , Proteínas Proto-Oncogênicas B-raf/genética , Proteínas Proto-Oncogênicas c-akt/genética , Substituição de Aminoácidos , Proliferação de Células , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Retroalimentação Fisiológica , Fibroblastos/citologia , Proteína Forkhead Box M1/metabolismo , Regulação da Expressão Gênica , Humanos , Mutação , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , Fosforilação , Cultura Primária de Células , Proteínas Proto-Oncogênicas B-raf/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de SinaisRESUMO
Objective: The purpose of this study was to investigate how Korean counselors modify the helping skills they learned during training in their work with Korean clients. Method: Thirteen practicing Korean therapists who had taken two master's level courses in the Hill helping skills model were interviewed about their experiences in applying the model in their work with clients. Data were analyzed using consensual qualitative research (CQR). Results: Several characteristics were noted of Korean clients that might influence the appropriateness of the helping skills model (e.g., clients regard counseling as authorities and wise experts). Participants had a number of reactions to the overall helping skills model (e.g., noting that Korean counselors deliver empathy and genuineness more through non-verbal rather than verbal channels). Participants also noted needed modifications to specific stages or skills in the model (e.g., using the insight stage less than the exploration stage). Discussion: Based on these findings, six guidelines are offered for modifying the Hill helping skills approach to fit the needs of Korean clients: Provide a pre-exploration stage to educate clients about approach, utilize indirect and non-verbal communication more than verbal communication, validate client's experiences, work cautiously with emotions, be cautious about using insight skills, and respond to clients' implicit communication when they ask for action.
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Aconselhamento/métodos , Comportamento de Ajuda , Psicoterapia , Adulto , Feminino , Humanos , Entrevistas como Assunto , Pessoa de Meia-Idade , Modelos Psicológicos , Psicoterapia/métodos , República da CoreiaRESUMO
The purpose of the study was to examine the factor structure, measurement invariance, and psychometric properties of a commonly used measure of perceived career barriers (The Perception of Barriers Scale; Luzzo & McWhirter, 2001) with racially diverse college women. The results supported a 9-factor structure for the Perception of Barriers Scale; configural, metric, and scalar invariance for the 9-factors were found with Asian, African American, Latina, and White college women. All groups of women of color reported higher career barriers due to racism and higher educational barriers due to racial discrimination than White women. The results also demonstrated potential differences in salient barriers for women of color (educational barriers due to lack of confidence/skills for Asian women, career barriers due to sexism and racism for African American women, and educational barriers due to financial concerns for Latina women). The reliability estimates of the subscales were satisfactory and support for criterion validity was demonstrated. The results highlighted the importance of measuring and identifying salient barriers for college women of color. Future research and practice implications for assessing and ameliorating salient barriers for college women are discussed. (PsycINFO Database Record
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Mobilidade Ocupacional , Etnicidade/psicologia , Percepção , Grupos Raciais/psicologia , Racismo/psicologia , Sexismo/psicologia , Adolescente , Feminino , Humanos , Racismo/tendências , Reprodutibilidade dos Testes , Sexismo/tendências , Estudantes/psicologia , Universidades/tendências , Adulto JovemRESUMO
Asian American women's (AAW's) mental health issues have received growing public attention; recent statistics suggest alarmingly high suicide rates among AAW. Yet, little research has examined the nuanced oppression that AAW face and the daily effects of compounded racism and sexism contributing to their mental health issues. Applying the intersectionality and microaggressions framework, we developed the Gendered Racial Microaggressions Scale for Asian American Women (GRMSAAW) using data collected from 564 AAW. Items were developed via literature review, focus group, and expert review. Exploratory (N = 304) and confirmatory (N = 260) factor analyses suggested a 4-factor structure and produced 22-item scales of frequency and stress appraisal with the following subscales: (a) Ascription of Submissiveness, (b) Assumption of Universal Appearance, (c) Asian Fetishism, and (d) Media Invalidation. Internal consistency estimates were .80 and above for frequency and stress appraisal scales, and the scales accounted for 52% and 60% of variance, respectively. Examination of a bifactor model containing one general factor and four group factors suggested that GRMSAAW could be represented unidimensionally (total scale score) for the purpose of applied measurement. Initial construct validity was established as GRMSAAW scores were associated with sexism, racial microaggressions, depressive symptoms, and internalized racism in ways consistent with theory. Implications for research and practice are discussed. (PsycINFO Database Record (c) 2018 APA, all rights reserved).
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Agressão/psicologia , Asiático/psicologia , Racismo/psicologia , Sexismo/psicologia , Inquéritos e Questionários/normas , Adolescente , Adulto , Idoso , Depressão/diagnóstico , Depressão/etnologia , Depressão/psicologia , Análise Fatorial , Feminino , Grupos Focais , Identidade de Gênero , Humanos , Pessoa de Meia-Idade , Racismo/etnologia , Reprodutibilidade dos Testes , Sexismo/etnologia , Fatores Socioeconômicos , Adulto JovemRESUMO
Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and (64)Cu. HT29 (hTERT+) and U2OS (hTERT-) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo.
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Imunoglobulina M/metabolismo , Peptídeos/química , Telomerase/metabolismo , Animais , Linhagem Celular Tumoral , Fluorescência , Humanos , Imunoglobulina M/química , Camundongos , Camundongos NusRESUMO
OBJECTIVES: The purpose of this study was to evaluate the characteristics and importance of superficial echogenic lesions around cranial sutures on neonatal cranial sonography. METHODS: We retrospectively reviewed the clinical records and neuroimaging studies of 40 neonates who had superficial echogenic lesions around sutures on neonatal cranial sonography. Magnetic resonance imaging (n = 18) and computed tomography (n = 2) were performed within 2 weeks after sonography. We correlated sonographic findings with computed tomographic and magnetic resonance imaging findings and analyzed them. We also evaluated the associated lesions, neurologic signs, and follow-up changes. RESULTS: Sonographically, the superficial echogenic lesions involved both sulci and perisulcal parenchyma in 39 neonates and were located in the frontal and parietal areas around the sagittal suture in 38 neonates. Magnetic resonance imaging revealed a pattern of hypoxic ischemic encephalopathy in 9 neonates, birth trauma in 3 neonates, a mixed pattern of hypoxic ischemic encephalopathy and trauma in 3 neonates, nonspecific single infarctions in 2 neonates, and lack of a defined lesion in 1 neonate. The associated lesions were subdural hemorrhage (n = 12), epidural hematoma (n = 4), germinal matrix hemorrhage (n = 3), intraventricular hemorrhage (n = 2), and periventricular leukomalacia (n = 1). All epidural hematomas were associated with scalp hematoma, and 2 patients had skull fractures. One neonate with epidural hematoma associated with a hypoxic ischemic encephalopathy pattern showed mild spasticity in both ankles until 16 months. CONCLUSIONS: Superficial echogenic lesions detected around cranial sutures on neonatal sonography may be an indicator of more serious intracranial lesions such as more extensive hypoxic ischemic encephalopathy and intracranial hematomas, including epidural hematoma.
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Traumatismos do Nascimento/diagnóstico por imagem , Suturas Cranianas/diagnóstico por imagem , Ecoencefalografia/métodos , Hematoma Epidural Craniano/diagnóstico por imagem , Hipóxia-Isquemia Encefálica/diagnóstico por imagem , Hemorragia Intracraniana Traumática/diagnóstico por imagem , Diagnóstico Diferencial , Feminino , Humanos , Recém-Nascido , Imageamento por Ressonância Magnética , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tomografia Computadorizada por Raios XRESUMO
The therapeutic efficacy of radioiodine (¹³¹I) therapy has been reported to be variable among cancer patients and even between metastatic regions in the same patients. Because the expression level of sodium iodide symporter (NIS) cannot reflect the efficacy of therapy, other strategies are required to predict the precise therapeutic effect of ¹³¹I therapy. In this research, we investigated the correlation between iodine (I) uptake, apoptosis imaging, and therapeutic efficacy. Two HT29 cell lines, cytomegalovirus (CMV)-NIS (or NIS+++) and TERT-NIS (or NIS+), were established by retroviral transfection. I uptake was estimated by I-uptake assay and gamma camera imaging. Apoptosis was evaluated by confocal microscopy and a Maestro fluorescence imaging system (CRi Inc., Woburn, MA) using ApoFlamma (BioACTs, Seoul, Korea), a fluorescent dye-conjugated apoptosis-targeting peptide 1 (ApoPep-1). Therapeutic efficacy was determined by tumor size. The CMV-NIS showed higher I uptake and ApoFlamma signals than TERT-NIS. In xenograft models, CMV-NIS also showed high 99m technetium signals and ApoFlamma signals. Tumor reduction had a stronger correlation with apoptosis imaging signals than with gamma camera imaging signals, which reflect I uptake. Higher NIS-expressing tumors showed increased apoptosis and I uptake, resulting in a significant tumor reduction. Moreover, tumor reduction showed a strong correlation with ApoFlamma imaging compared to I-uptake imaging.
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Neoplasias do Colo/radioterapia , Radioisótopos do Iodo/farmacocinética , Oligopeptídeos/farmacologia , Simportadores/metabolismo , Tecnécio/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Terapia Genética , Vetores Genéticos/administração & dosagem , Células HT29 , Humanos , Radioisótopos do Iodo/uso terapêutico , Microscopia Confocal , Oligopeptídeos/química , Imagem Óptica , Retroviridae/genética , Simportadores/genética , Resultado do TratamentoRESUMO
Since therapy-induced senescence (TIS) can either support or inhibit cancer progression, identifying which types of chemotherapeutic agents can produce the strongest anti-tumor TIS is an important issue. Here, cyclin-dependent kinase4/6 inhibitors (CDK4/6i)-induced senescence was compared to the TIS induced by conventional DNA-damaging agents. Despite both types of agents eliciting a similar degree of senescence, we observed increased expression of the senescence-associated secretory phenotype (SASP) and ligands related to pro-tumor immunity (IL6, CXCL8, TGFß, CD274, and CEACAM1) and angiogenesis (VEGFA) mainly in TIS induced by DNA-damaging agents rather than by CDK4/6i. Additionally, although all agents increased the expression of anti-tumor immunomodulatory proteins related to antigen presentation (MHC-I, B2M) and T cell chemokines (CXCL9, 10, 11), CDK4/6i-induced senescent cells still maintained this expression at a similar or even higher intensity than cells treated with DNA-damaging agents, despite the absence of nuclear factor-kappa-B (NF-κB) and p53 activation. These data suggest that in contrast with DNA-damaging agents, which augment the pro-tumorigenic microenvironment via pro-inflammatory SASP, CDK4/6i can generate TIS only with antitumor immunomodulatory proteins.
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Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , NF-kappa B/metabolismo , Senescência Celular/genética , Microambiente Tumoral , Quinase 4 Dependente de CiclinaRESUMO
In animal farming, timely estrus detection and prediction of the best moment for insemination is crucial. Traditional sow estrus detection depends on the expertise of a farm attendant which can be inconsistent, time-consuming, and labor-intensive. Attempts and trials in developing and implementing technological tools to detect estrus have been explored by researchers. The objective of this review is to assess the automatic methods of estrus recognition in operation for sows and point out their strong and weak points to assist in developing new and improved detection systems. Real-time methods using body and vulvar temperature, posture recognition, and activity measurements show higher precision. Incorporating artificial intelligence with multiple estrus-related parameters is expected to enhance accuracy. Further development of new systems relies mostly upon the improved algorithm and accurate data provided. Future systems should be designed to minimize the misclassification rate, so better detection is achieved.
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In this study, we explore the dynamic process of colorectal cancer progression, emphasizing the evolution toward a more metastatic phenotype. The term "evolution" as used in this study specifically denotes the phenotypic transition toward a higher metastatic potency from well-formed glandular structures to collective invasion, ultimately resulting in the development of cancer cell buddings at the invasive front. Our findings highlight the spatial correlation of this evolution with tumor cell senescence, revealing distinct types of senescent tumor cells (types I and II) that play different roles in the overall cancer progression. Type I senescent tumor cells (p16INK4A+/CXCL12+/LAMC2-/MMP7-) are identified in the collective invasion region, whereas type II senescent tumor cells (p16INK4A+/CXCL12+/LAMC2+/MMP7+), representing the final evolved form, are prominently located in the partial-EMT region. Importantly, type II senescent tumor cells associate with local invasion and lymph node metastasis in colorectal cancer, potentially affecting patient prognosis.
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Neoplasias Colorretais , Metaloproteinase 7 da Matriz , Humanos , Metaloproteinase 7 da Matriz/genética , Senescência Celular/genética , Fenótipo , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologiaRESUMO
Forkhead Box M1 (FoxM1) protein is a transcription factor and regulates cell cycle. It is commonly upregulated in human cancer tissue and correlated with poor prognosis, suggesting that the overexpression of FoxM1 plays a critical role in carcinogenesis. In this study, we report the identification and characterization of a new variant of FoxM1, which was first isolated from our laboratory in hepatoma cell lines. Compared with wild-type FoxM1, the new variant lacks of C-terminus of FoxM1 (FoxM1ΔC), which is a transactivation domain. Reverse transcription-polymerase chain reaction and western blot analysis demonstrated that FoxM1ΔC was highly expressed in a variety of cancer cell lines such as HepG2, HeLa, A549, MB231, EJ, U2OS, Hep3B and MCF7, but not expressed in normal human dermal fibroblast (HDF). Immunoprecipitation assay revealed that FoxM1ΔC interacted with wild-type FoxM1. Furthermore, FoxM1ΔC bound to FoxM1 targeted gene promoter region and correlated with dysregulation of wild-type FoxM1. FoxM1ΔC delayed G2/M to G1 progression of cell cycle, decreased Aurora B(T232) phosphorylation and increased chromosome centromere interspace. Finally, FoxM1ΔC induced instability of chromosome and formation of aneuploid cells within 1 month when expressed in HDF. In conclusion, FoxM1ΔC is expressed in cancer cells and dysregulates normal cell cycle and induces chromosome instability.
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Instabilidade Cromossômica , Fatores de Transcrição Forkhead/biossíntese , Fatores de Transcrição Forkhead/genética , Ciclo Celular/genética , Linhagem Celular Tumoral , Proteína Forkhead Box M1 , Fatores de Transcrição Forkhead/metabolismo , Regulação Neoplásica da Expressão Gênica , Células HeLa , Células Hep G2 , Humanos , Células MCF-7 , Regiões Promotoras Genéticas , Transcrição Gênica , Ativação TranscricionalRESUMO
Gallium-68-labeled 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA)-cyclic Arg-Gly-Asp-D-Tyr-Lys (c(RGDyK)) was developed for αvß3 targeting, and is a promising agent for imaging of cancer and disorders related to angiogenesis. In this study, we performed kinetic analysis of (68)Ga-NOTA-c(RGDyK) in rats with surgically induced forelimb ischemia, and immunohistochemical analysis was also performed to assess αvß3 immuno-staining level. Animal models were created by excision of the left brachial vessels, and a sham operation was performed on the right brachial region under 2 % isoflurane anesthesia. Using an animal positron emission tomography/computed tomography (PET/CT) scanner, a list mode PET scan (120 min) was started with the injection of (68)Ga-NOTA-c(RGDyK) via the tail vein at 3, 5 and 7 days after ischemic surgery. Volumes of interest were drawn on the left ventricle, sham operation, control, and ischemic regions. Compartmental and two graphical analyses (Logan and RE plots) were performed for kinetic parameter estimation. The immunohistochemical analysis was also performed after the last PET scan, and cell components were scored on a six point scale for quantification of immuno-staining level (0-negative to 5-very high). A 3-compartment model with reversible binding best described the tissue time-activity curves. The distribution volume of the ischemic region was significantly higher than that of the sham operation (P < 10(-6)) and control region (P < 10(-9)). Both the Logan and RE plots showed high correlation with compartmental analysis (R(2) = 0.96 and 0.95 for Logan and RE, respectively). The temporal changes in distribution volume and binding potential were not significant. The immuno-staining level of the ischemic region was significantly higher than that of sham operation (P < 10(-4)) and control region (P < 10(-8)). Kinetic modeling studies with dynamic (68)Ga-NOTA-c(RGDyK) PET scan are feasible based on an image-derived input function in a rat ischemia model. The kinetic modeling analysis performed in this study will be useful for the quantitative evaluation of (68)Ga-NOTA-c(RGDyK) binding to αvß3 in angiogenic tissues.
Assuntos
Meios de Contraste , Endotélio Vascular/química , Membro Anterior/irrigação sanguínea , Integrina alfaVbeta3/análise , Isquemia/diagnóstico por imagem , Imagem Multimodal/métodos , Neovascularização Fisiológica , Compostos Organometálicos , Peptídeos Cíclicos , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos , Tomografia Computadorizada por Raios X/métodos , Animais , Meios de Contraste/farmacocinética , Endotélio Vascular/fisiologia , Desenho de Equipamento , Membro Anterior/diagnóstico por imagem , Imuno-Histoquímica , Isquemia/fisiopatologia , Miniaturização , Modelos Animais , Imagem Multimodal/instrumentação , Imagem Multimodal/veterinária , Músculo Esquelético/irrigação sanguínea , Compostos Organometálicos/farmacocinética , Peptídeos Cíclicos/farmacocinética , Tomografia por Emissão de Pósitrons/instrumentação , Tomografia por Emissão de Pósitrons/veterinária , Compostos Radiofarmacêuticos/farmacocinética , Ratos , Ratos Sprague-Dawley , Tomografia Computadorizada por Raios X/instrumentação , Tomografia Computadorizada por Raios X/veterinária , CicatrizaçãoRESUMO
Rotenone is an inhibitor of mitochondrial complex I-induced neurotoxicity in PC12 cells and has been widely studied to elucidate the pathogenesis of Parkinson's disease. We investigated the neuroprotective effects of betaine on rotenone-induced neurotoxicity in PC12 cells. Betaine inhibited rotenone-induced apoptosis in a dose-dependent manner, with cell viability increasing from 50 % with rotenone treatment alone to 71 % with rotenone plus 100-µM betaine treatment. Flow cytometric analysis demonstrated cell death in the rotenone-treated cells to be over 50 %; the number of live cells increased with betaine pretreatment. Betaine pretreatment of PC12 cells attenuated rotenone-mediated mitochondrial dysfunction, including nuclear fragmentation, ATP depletion, mitochondrial membrane depolarization, caspase-3/7 activation, and reactive oxygen species production. Western blots demonstrated activation of caspase-3 and caspase-9, and their increased expression levels in rotenone-treated cells; betaine decreased caspase-3 and caspase-9 expression levels and suppressed their activation. Together, these results suggest that betaine may serve as a neuroprotective agent in the treatment of neurodegenerative diseases.
Assuntos
Betaína/farmacologia , Fármacos Neuroprotetores/farmacologia , Neurotoxinas/toxicidade , Rotenona/toxicidade , Animais , Caspases/metabolismo , Metabolismo Energético/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Citometria de Fluxo , Marcação In Situ das Extremidades Cortadas , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Microscopia Confocal , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Células PC12 , Ratos , Superóxidos/metabolismoRESUMO
Structure-activity relationship (SAR) calculations were used to find monoamine oxidase-B (MAO-B) inhibitors by identifying pharmacophores exhibiting high inhibitory activities. Several such chromenylchalcones were designed and synthesized accordingly. Their inhibitory effects on MAO-B were determined using an HPLC-based method and an MAO-B enzyme assay kit. (E)-3-(6-Methoxy-2H-chromen-3-yl)-1-(2-methoxyphenyl)prop-2-en-1-one exhibited a half-maximal inhibitory concentration of 320 nM. Its molecular-level binding mode with the three-dimensional structure of MAO-B was elucidated using an in silico docking study. The chromenylchalcone scaffold, which is derived from natural products including isoflavonoids and chalcones, had not been previously reported as an MAO-B inhibitor.
Assuntos
Benzopiranos/farmacologia , Chalconas/farmacologia , Inibidores da Monoaminoxidase/farmacologia , Monoaminoxidase/metabolismo , Benzopiranos/química , Chalconas/química , Relação Dose-Resposta a Droga , Humanos , Modelos Moleculares , Estrutura Molecular , Monoaminoxidase/química , Inibidores da Monoaminoxidase/química , Relação Estrutura-AtividadeRESUMO
Chalcones are of interest to medicinal chemists because their structures can be easily modified with various functional groups. The syntheses and biological activities of chalcones from natural sources are well known. In this study, 24 2'-hydroxychalcones bearing methoxy substituents were synthesized, among which five are new. The NMR data for all synthesized chalcones are described for the first time. The complete assignments of the (1)H and (13)C NMR data can be used for the identification of newly discovered and widely isolated, synthesized chalcones.
Assuntos
Chalconas/química , Isótopos de Carbono , Espectroscopia de Ressonância Magnética/normas , Prótons , Padrões de ReferênciaRESUMO
The late gestation period is crucial for fetal growth and development, impacting swine enterprises' profitability. Various nutritional strategies have been explored to enhance reproductive performance in sows, but findings regarding birth weight and litter size have been inconsistent. This study investigated the effects of increased feeding allowance during the late gestation period on the reproductive performance and farrowing behavior of primiparous and multiparous sows. A total of 28 sows (Landrace × Yorkshire) were used in this experiment, and fed 2.50 kg/d or 3.50 kg/d from 84 days of gestation until farrowing. Farrowing behavior was monitored using a DeepEyesTM M3SEN camera. The data were analyzed using the 2 × 2 factorial within Statistical Analysis System (SAS, 2011, Version 9.3) software. The results indicated that regardless of the parity number, sows fed a high diet exhibited a numerical increase in the total number of born piglets and a significant increase in milk yield (p = 0.014) and piglet birthweight (p = 0.023). Backfat thickness loss was significantly higher in sows with a 2.50 kg feeding allowance (p = 0.022), and the total number of piglets born, live births, and litter size were numerically higher in sows fed 3.50 kg per day. Moreover, stillborn piglets, mortality rate, and re-estrus days were numerically lower in sows with a high feeding allowance. The diet and parity did not individually affect the average duration of farrowing and farrowing intervals. However, the duration of postural changes in sows after farrowing was significantly reduced (p = 0.012). The principal component analysis revealed 81.40% and 80.70% differences upon partial least-squares discriminant analysis. Therefore, increasing feeding allowance during the late gestation period, regardless of parity, could positively influence sows' reproductive performance and piglets' growth performance during the lactation phase.