RESUMO
The fungal genus Trichoderma is a rich source of structurally diverse secondary metabolites with remarkable pharmaceutical properties. The chemical constituents and anticancer activities of the marine-derived fungus Trichoderma lixii have never been investigated. In this study, a bioactivity-guided investigation led to the isolation of eleven compounds, including trichodermamide A (1), trichodermamide B (2), aspergillazine A (3), DC1149B (4), ergosterol peroxide (5), cerebrosides D/C (6/7), 5-hydroxy-2,3-dimethyl-7-methoxychromone (8), nafuredin A (9), and harzianumols E/F (10/11). Their structures were identified by using various spectroscopic techniques and compared to those in the literature. Notably, compounds 2 and 5-11 were reported for the first time from this species. Evaluation of the anticancer activities of all isolated compounds was carried out. Compounds 2, 4, and 9 were the most active antiproliferative compounds against three cancer cell lines (human myeloma KMS-11, colorectal HT-29, and pancreas PANC-1). Intriguingly, compound 4 exhibited anti-austerity activity with an IC50 of 22.43 µM against PANC-1 cancer cells under glucose starvation conditions, while compound 2 did not.
Assuntos
Antineoplásicos , Trichoderma , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Humanos , Trichoderma/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Estrutura Molecular , Organismos Aquáticos/química , Ensaios de Seleção de Medicamentos AntitumoraisRESUMO
PURPOSE: Common dense stereo simultaneous localization and mapping (SLAM) approaches in minimally invasive surgery (MIS) require high-end parallel computational resources for real-time implementation. Yet, it is not always feasible since the computational resources should be allocated to other tasks like segmentation, detection, and tracking. To solve the problem of limited parallel computational power, this research aims at a lightweight dense stereo SLAM system that works on a single-core CPU and achieves real-time performance (more than 30 Hz in typical scenarios). METHODS: A new dense stereo mapping module is integrated with the ORB-SLAM2 system and named BDIS-SLAM. Our new dense stereo mapping module includes stereo matching and 3D dense depth mosaic methods. Stereo matching is achieved with the recently proposed CPU-level real-time matching algorithm Bayesian Dense Inverse Searching (BDIS). A BDIS-based shape recovery and a depth mosaic strategy are integrated as a new thread and coupled with the backbone ORB-SLAM2 system for real-time stereo shape recovery. RESULTS: Experiments on in vivo data sets show that BDIS-SLAM runs at over 30 Hz speed on modern single-core CPU in typical endoscopy/colonoscopy scenarios. BDIS-SLAM only consumes around an additional 12 % time compared with the backbone ORB-SLAM2. Although our lightweight BDIS-SLAM simplifies the process by ignoring deformation and fusion procedures, it can provide a usable dense mapping for modern MIS on computationally constrained devices. CONCLUSION: The proposed BDIS-SLAM is a lightweight stereo dense SLAM system for MIS. It achieves 30 Hz on a modern single-core CPU in typical endoscopy/colonoscopy scenarios (image size around 640 × 480 ). BDIS-SLAM provides a low-cost solution for dense mapping in MIS and has the potential to be applied in surgical robots and AR systems. Code is available at https://github.com/JingweiSong/BDIS-SLAM .
Assuntos
Algoritmos , Imageamento Tridimensional , Procedimentos Cirúrgicos Minimamente Invasivos , Humanos , Imageamento Tridimensional/métodos , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Procedimentos Cirúrgicos Minimamente Invasivos/instrumentação , Cirurgia Assistida por Computador/métodos , Teorema de BayesRESUMO
Colorectal cancer accounted for the third most common cancer in the world. The search for new drug candidates that can be used for colorectal cancer treatment from marine-derived fungi, Emericella sp. The present study was performed to isolate the cytotoxic compound from Emericella sp. The isolation method was carried out by using a combination of chromatographic techniques to afford compound 1. The cytotoxic activity and the exosome production property were determined by using proliferation and luciferase assay against HT29 CD63 Nluc cells, respectively. The chemical structure of compound 1 was identified as cordycepin based on spectroscopy methods such as mass spectrometry and nuclear magnetic resonance (1D and 2D NMR) analyses and comparison with authentic spectral data. The biological activity assay showed that cordycepin exhibited cytotoxic activity with an IC50 value of 92.05 µM through proliferation assay, and also inhibited the exosome production by luciferase assay with an IC50 value of 86.47 µM. Cordycepin was isolated from culture broth Emericella sp., exhibiting moderate cytotoxic activity and inhibitory activity of exosome production. Thus, cordycepin is a potential compound to be investigated further for its exosome production inhibition activity for further use as an anticancer lead compound.
Assuntos
Antineoplásicos , Neoplasias do Colo , Neoplasias Colorretais , Emericella , Humanos , Emericella/química , Aspergillus , Linhagem Celular Tumoral , Fungos , Neoplasias do Colo/tratamento farmacológico , Luciferases , Estrutura Molecular , Antineoplásicos/químicaRESUMO
This study estimates the association between sarcopenia and blood biochemical parameters, nutritional intake, anthropometric measurements, physical performance, and physical activity in patients with type 2 diabetes mellitus (T2DM). Participants were recruited from a primary care clinic in Kaohsiung City. According to the diagnosis criteria of the Asian Working Group for Sarcopenia (AWGS) in 2019, 110 patients with T2DM (aged 50-80 years) were divided into three groups: non-sarcopenia (n = 38), possible sarcopenia (n = 31), and sarcopenia (n = 41). Blood samples were collected, and nutritional intake was evaluated by a registered dietitian. A food frequency questionnaire and a Godin leisure-time exercise questionnaire were used to assess their daily vitamin D intake and physical activity. There were significant differences in age, serum vitamin D levels, nutritional intake, anthropometric measurements, and physical performance between the three groups. In elderly patients with T2DM, reduced serum 25-hydroxyvitamin D [25(OH)D] levels and daily energy intake were significantly associated with possible sarcopenia. Age, lower BMI, reduced serum 25(OH)D, and reduced dietary protein and vitamin D intake were significantly associated with sarcopenia. These findings may serve as the basis for intervention trials to reduce the prevalence of sarcopenia.
Assuntos
Diabetes Mellitus Tipo 2 , Sarcopenia , Idoso , Humanos , Diabetes Mellitus Tipo 2/complicações , Taiwan/epidemiologia , Sarcopenia/epidemiologia , Sarcopenia/etiologia , Composição Corporal , Ingestão de Alimentos , Exercício Físico , Vitamina DRESUMO
Cancer cells secrete aberrantly large amounts of extracellular vesicles (EVs) including exosomes, which originate from multivesicular bodies (MVBs). Because EVs potentially contribute to tumor progression, EV inhibitors are of interest as novel therapeutics. We screened a fungal natural product library. Using cancer cells engineered to secrete luciferase-labeled EVs, we identified asteltoxin, which inhibits mitochondrial ATP synthase, as an EV inhibitor. Low concentrations of asteltoxin inhibited EV secretion without inducing mitochondrial damage. Asteltoxin attenuated cellular ATP levels and induced AMPK-mediated mTORC1 inactivation. Consequently, MiT/TFE transcription factors are translocated into the nucleus, promoting transcription of lysosomal genes and lysosome activation. Electron microscopy analysis revealed that the number of lysosomes increased relative to that of MVBs and the level of EVs decreased after treatment with asteltoxin or rapamycin, an mTORC1 inhibitor. These findings suggest that asteltoxin represents a new type of EV inhibitor that controls MVB fate.
Assuntos
Proteínas Quinases Ativadas por AMP , Vesículas Extracelulares , Lisossomos , Alvo Mecanístico do Complexo 1 de Rapamicina , Pironas , Serina-Treonina Quinases TORRESUMO
A concise synthesis of cajaninstilbene acid was achieved in 7 steps from (E)-3,5-dimethoxystilbene in 8.6% overall yield via the Claisen rearrangement of an aryl reverse-prenyl ether as the key step. Cytotoxic activities against human pancreatic carcinoma PANC-1 cells of cajaninstilbene acid and amorfrutins A-D were also evaluated.
Assuntos
Citotoxinas , Estilbenos , Humanos , Neoplasias Pancreáticas , Salicilatos , Estilbenos/farmacologia , Neoplasias PancreáticasRESUMO
BACKGROUND: Intestinal barrier breakdown, a frequent complication of intestinal ischemia-reperfusion (I/R) including dysfunction and the structure changes of the intestine, is characterized by a loss of tight junction and enhanced permeability of the intestinal barrier and increased mortality. To develop effective and novel therapeutics is important for the improvement of outcome of patients with intestinal barrier deterioration. Recombinant human angiopoietin-like protein 4 (rhANGPTL4) is reported to protect the blood-brain barrier when administered exogenously, and endogenous ANGPTL4 deficiency deteriorates radiation-induced intestinal injury. AIM: To identify whether rhANGPTL4 may protect intestinal barrier breakdown induced by I/R. METHODS: Intestinal I/R injury was elicited through clamping the superior mesenteric artery for 60 min followed by 240 min reperfusion. Intestinal epithelial (Caco-2) cells and human umbilical vein endothelial cells were challenged by hypoxia/ reoxygenation to mimic I/R in vitro. RESULTS: Indicators including fluorescein isothiocyanate-conjugated dextran (4 kilodaltons; FD-4) clearance, ratio of phosphorylated myosin light chain/total myosin light chain, myosin light chain kinase and loss of zonula occludens-1, claudin-2 and VE-cadherin were significantly increased after intestinal I/R or cell hypoxia/reoxygenation. rhANGPTL4 treatment significantly reversed these indicators, which were associated with inhibiting the inflammatory and oxidative cascade, excessive activation of cellular autophagy and apoptosis and improvement of survival rate. Similar results were observed in vitro when cells were challenged by hypoxia/reoxygenation, whereas rhANGPTL4 reversed the indicators close to normal level in Caco-2 cells and human umbilical vein endothelial cells significantly. CONCLUSION: rhANGPTL4 can function as a protective agent against intestinal injury induced by intestinal I/R and improve survival via maintenance of intestinal barrier structure and functions.
Assuntos
Proteína 4 Semelhante a Angiopoietina/farmacologia , Intestinos , Traumatismo por Reperfusão , Células CACO-2 , Células Endoteliais da Veia Umbilical Humana , Humanos , Mucosa Intestinal , Proteínas Recombinantes/farmacologia , Traumatismo por Reperfusão/prevenção & controleRESUMO
Graded index (GRIN) lenses are commonly used for compact imaging systems. It is not widely appreciated that the ion-exchange process that creates the rotationally symmetric GRIN lens index profile also causes a symmetric birefringence variation. This property is usually considered a nuisance, such that manufacturing processes are optimized to keep it to a minimum. Here, rather than avoiding this birefringence, we understand and harness it by using GRIN lenses in cascade with other optical components to enable extra functionality in commonplace GRIN lens systems. We show how birefringence in the GRIN cascades can generate vector vortex beams and foci, and how it can be used advantageously to improve axial resolution. Through using the birefringence for analysis, we show that the GRIN cascades form the basis of a new single-shot Müller matrix polarimeter with potential for endoscopic label-free cancer diagnostics. The versatility of these cascades opens up new technological directions.
Assuntos
Endoscopia/instrumentação , Lentes , Neoplasias/diagnóstico por imagem , Óptica e Fotônica , Polarimetria de Varredura a Laser/instrumentação , Birrefringência , Neoplasias/diagnóstico , Refratometria , Polarimetria de Varredura a Laser/métodosRESUMO
Sustained-release systems made by biodegradable polymers for protein and peptide drug delivery have received considerable attention by academic researchers and major pharmaceutical companies around the world. Various types of biodegradable materials, including natural and synthetic polymers, have been applied to form protein and peptide drug carriers. Among these material candidates, poly lactic acid (PLA) and poly lactic-co-glycolic acid (PLGA) are the most commonly used biodegradable materials in the development of protein and peptide microspheres. In addition, many microsphere preparation technologies, including spray drying, coacervation, multiple emulsion solvent evaporation method and microporous membrane emulsification have been developed for microspheres preparation. In this review, we particularly summarize and briefly introduce the materials and methods that are used to fabricate microspheres as protein delivery systems. The existing opportunities and challenges for successful protein delivery are also discussed.
Assuntos
Portadores de Fármacos/química , Microesferas , Peptídeos/química , Polímeros/química , Proteínas/química , Preparações de Ação Retardada/química , Composição de Medicamentos , Microfluídica/métodosRESUMO
PURPOSE: Intra-operative measurement of tissue oxygen saturation ([Formula: see text]) is important in detection of ischaemia, monitoring perfusion and identifying disease. Hyperspectral imaging (HSI) measures the optical reflectance spectrum of the tissue and uses this information to quantify its composition, including [Formula: see text]. However, real-time monitoring is difficult due to capture rate and data processing time. METHODS: An endoscopic system based on a multi-fibre probe was previously developed to sparsely capture HSI data (sHSI). These were combined with RGB images, via a deep neural network, to generate high-resolution hypercubes and calculate [Formula: see text]. To improve accuracy and processing speed, we propose a dual-input conditional generative adversarial network, Dual2StO2, to directly estimate [Formula: see text] by fusing features from both RGB and sHSI. RESULTS: Validation experiments were carried out on in vivo porcine bowel data, where the ground truth [Formula: see text] was generated from the HSI camera. Performance was also compared to our previous super-spectral-resolution network, SSRNet in terms of mean [Formula: see text] prediction accuracy and structural similarity metrics. Dual2StO2 was also tested using simulated probe data with varying fibre number. CONCLUSIONS: [Formula: see text] estimation by Dual2StO2 is visually closer to ground truth in general structure and achieves higher prediction accuracy and faster processing speed than SSRNet. Simulations showed that results improved when a greater number of fibres are used in the probe. Future work will include refinement of the network architecture, hardware optimization based on simulation results, and evaluation of the technique in clinical applications beyond [Formula: see text] estimation.
Assuntos
Intestinos/diagnóstico por imagem , Isquemia/diagnóstico por imagem , Imagem Óptica/métodos , Oxigênio , Animais , SuínosRESUMO
OBJECTIVE: At present, there are no objective techniques to quantify and describe laryngeal obstruction, and the reproducibility of subjective manual quantification methods is insufficient, resulting in diagnostic inaccuracy and a poor signal-to-noise ratio in medical research. In this work, a workflow is proposed to quantify laryngeal movements from laryngoscopic videos, to facilitate the diagnosis procedure. METHODS: The proposed method analyses laryngoscopic videos, and delineates glottic opening, vocal folds, and supraglottic structures, using a convolutional neural networks (CNNs) based algorithm. The segmentation is divided into two steps: A bounding box which indicates the region of interest (RoI) is found, followed by segmentation using fully convolutional networks (FCNs). The segmentation results are statistically quantified along the temporal dimension and processed using singular spectrum analysis (SSA), to extract clear objective information that can be used by the clinicians in diagnosis. RESULTS: The segmentation was validated on 400 images from 20 videos acquired using different endoscopic systems from different patients. The results indicated significant improvements over using FCN only in terms of both processing speed (16 FPS vs. 8 FPS) and segmentation result statistics. Five clinical cases on patients have also been provided to showcase the quantitative analysis results using the proposed method. CONCLUSION: The proposed method guarantees a robust and fast processing of laryngoscopic videos. Measurements of glottic angles and supraglottic index showed distinctive patterns in the provided clinical cases. SIGNIFICANCE: The proposed automated and objective method extracts important temporal laryngeal movement information, which can be used to aid laryngeal closure diagnosis.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Laringoscopia/métodos , Laringe/diagnóstico por imagem , Redes Neurais de Computação , Processamento de Sinais Assistido por Computador , Algoritmos , Humanos , Laringe/fisiologia , Reprodutibilidade dos Testes , Gravação em VídeoRESUMO
Surgical guidance and decision making could be improved with accurate and real-time measurement of intra-operative data including shape and spectral information of the tissue surface. In this work, a dual-modality endoscopic system has been proposed to enable tissue surface shape reconstruction and hyperspectral imaging (HSI). This system centers around a probe comprised of an incoherent fiber bundle, whose fiber arrangement is different at the two ends, and miniature imaging optics. For 3D reconstruction with structured light (SL), a light pattern formed of randomly distributed spots with different colors is projected onto the tissue surface, creating artificial texture. Pattern decoding with a Convolutional Neural Network (CNN) model and a customized feature descriptor enables real-time 3D surface reconstruction at approximately 12 frames per second (FPS). In HSI mode, spatially sparse hyperspectral signals from the tissue surface can be captured with a slit hyperspectral imager in a single snapshot. A CNN based super-resolution model, namely "super-spectral-resolution" network (SSRNet), has also been developed to estimate pixel-level dense hypercubes from the endoscope cameras standard RGB images and the sparse hyperspectral signals, at approximately 2 FPS. The probe, with a 2.1â¯mm diameter, enables the system to be used with endoscope working channels. Furthermore, since data acquisition in both modes can be accomplished in one snapshot, operation of this system in clinical applications is minimally affected by tissue surface movement and deformation. The whole apparatus has been validated on phantoms and tissue (ex vivo and in vivo), while initial measurements on patients during laryngeal surgery show its potential in real-world clinical applications.
Assuntos
Endoscópios , Tecnologia de Fibra Óptica , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional , Redes Neurais de Computação , Algoritmos , Humanos , Período Intraoperatório , Análise Espacial , Análise EspectralRESUMO
OBJECTIVE: Intestinal perforation is a rare complication after liver transplantation. This study was designed to calculate the incidence and investigate the outcomes of intestinal perforation in adult liver transplant patients. MATERIALS AND METHODS: The clinical records of liver transplant recipients between January 2014 and June 2016 were obtained. The incidence of intestinal perforation was calculated, and high risk factors were analyzed. RESULTS: The mean operative time was 8.5 h (range: 6-11 h). The mean portal vein occlusion time was 66.5 min (range: 58-72 min), and the mean cold ischemia time was 7.9 h (range: 6.5-9.5 h). Four (2.7%) patients developed intestinal perforation from 9 to 14 days postliver transplant. All perforations were single and repaired by interrupted silk sutures. Two patients uneventfully recovered, but intestinal perforation recurred in two other patients. Simple repair was undertaken in one patient, and terminal ileum resection and ileostomy were performed in the other patient. There were no perioperative deaths. CONCLUSION: The incidence of intestinal perforation after liver transplantation is low. Prompt diagnosis and treatment should be carried out to reduce comorbidities and mortality.
RESUMO
BACKGROUND: The management of obstructive left colon cancer (OLCC) remains debatable with the single-stage procedure of primary colonic anastomosis after cancer resection and on-table intracolonic lavage now being supported. PATIENTS AND METHODS: Patients with acute OLCC who were admitted between January 2008 and January 2015 were distributed into 5 different groups. Group ICI underwent emergency laparotomy for primary anastomosis following colonic resection and intraoperative colonic lavage; Group HP underwent emergency Hartmann's Procedure; Group CON consisted of patients treated by conservative management with subsequent elective open cancer resection; Group COL were colostomy patients; and Group INT consisted of patients who had interventional radiology followed by open elective colon cancer resection. The demographics of the patients and comorbidity, intraoperative data, and postoperative data were collected, with P < .05 as significant. RESULTS: There were 4 deaths in 138 cases (2.90%). There was only 1 patient who had anastomotic leakage (5.56%) in Group ICI, compared with none in Group HP and Group COL, 1 case in Group INT (7.69%), and 2 cases in Group CON (6.06%) (P > .05). Group INT and Group CON, when compared to the three surgical groups, Groups ICI, Group COL, and Group HP, individually, were statistically significant for the duration of surgery (P < .05). CONCLUSIONS: Primary anastomosis following colonic resection after irrigation can be safely performed in selected patients, with the necessary surgical expertise, with no increased risk in mortality, anastomotic leakage, and other postoperative complications.
Assuntos
Anastomose Cirúrgica/métodos , Neoplasias do Colo/cirurgia , Obstrução Intestinal/etiologia , Irrigação Terapêutica/métodos , Idoso , Idoso de 80 Anos ou mais , Fístula Anastomótica/epidemiologia , Neoplasias do Colo/diagnóstico , Colostomia/métodos , Feminino , Humanos , Obstrução Intestinal/cirurgia , Cuidados Intraoperatórios/métodos , Laparotomia/métodos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologiaRESUMO
Liver ischemia and reperfusion (I/R) injury is characterized by defective liver autophagy accompanied by alterations to the endogenous defense system. Dihydromyricetin (DHM) is a natural flavonoid that demonstrates a wide range of physiological functions, and has been implicated as a regulator of autophagy. This study investigates the protective effects of DHM pretreatment on liver injury caused by ischemia/reperfusion (I/R) and elucidates the potential mechanism of DHM-mediated protection. Mice were subjected to 60 minutes of ischemia followed by 5 hours of reperfusion. DHM (100 mg/kg bw/day) or the vehicle was administered daily by gavage 7 days before ischemia and immediately before reperfusion. In this study, DHM markedly decreased serum aminotransferase activity and inhibited liver I/R -stimulated apoptosis. Moreover, DHM exerted hepatoprotective effects by upregulating mRNA levels of various essential autophagy-related genes including ATG5, ATG12, BECN1, and LC3. Autophagy inhibitor chloroquine or Atg5 knockdown blocked DHM -mediated elevation in liver function. Specifically, DHM significantly increased FOXO3a expression, and enhanced FOXO3a nuclear translocation and Ser588 phosphorylation modification. Importantly, the inhibition of FOXO3a with FOXO3a-siRNA in mice decreased DHM-induced autophagy-related genes and diminished the protective effects of DHM against liver I/R injury. In summary, these findings identify DHM as a novel hepatoprotective small molecule by elevating FOXO3a expression and nuclear translocation, stimulating autophagy-related genes and suppressing liver I/R-induced apoptosis, suggesting FOXO3a may have therapeutic value in liver cell protection in liver I/R injury.
Assuntos
Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Flavonóis/farmacologia , Proteína Forkhead Box O3/genética , Hepatopatias/metabolismo , Substâncias Protetoras/farmacologia , Traumatismo por Reperfusão/metabolismo , Animais , Caspase 3/metabolismo , Modelos Animais de Doenças , Proteína Forkhead Box O3/metabolismo , Hepatopatias/tratamento farmacológico , Hepatopatias/patologia , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/patologiaRESUMO
This study was designed to investigate the relationship between prognosis of pancreatic head cancer and status of para-aortic lymph node (PALN). A total of 233 patients with pancreatic head cancer who underwent surgical resection between February 2008 and October 2015 were enrolled in this study. Univariate and multivariate analyses were used to reveal the prognostic factors. Prognostic factors for patients with and without metastasis of PALN were analyzed, respectively. The 5-year overall survival (OS) rate was 19.0% for all patients, and the positive rate of PALN metastasis was 18.9% (44/233). The 1-, 2-, 3-, and 5-year OS rates in patients without metastasis of PALN were 79.4%, 54.8%, 36.4%, and 22.9%, respectively, whereas the 1-, 2-, and 3-year survival rates were 54.0%, 14.8%, and 0%, respectively, in patients with metastasis of PALN. Preoperative CA19-9 level, tumor size, T status, N status, and adjuvant therapy were independent prognostic factors for all patients confirmed by multivariate analysis. For patients without PALN metastasis, back pain, tumor size, T status, N status, portal or superior mesenteric vein invasion, and adjuvant therapy were independent prognostic factors, while the only one influence factor for 2-year OS was adjuvant therapy for patients with metastasis of PALN. Metastasis of PALN was associated with poor prognosis for patients with pancreatic head cancer. Patients with and without metastasis of PALN had different prognostic factors, and adjuvant therapy was the only prognostic factor for patients with metastasis of PALN.
Assuntos
Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia , Adulto , Idoso , Quimioterapia Adjuvante , Intervalo Livre de Doença , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/tratamento farmacológico , Prognóstico , Fatores de Risco , Análise de Sobrevida , Resultado do TratamentoRESUMO
PURPOSE: In clinical examinations, the tissue surface topology is an important feature for detecting the tissue pathology and implementing augmented reality. We have previously presented a miniaturised structured light (SL) system for recovery of tissue surface shape in minimally invasive surgery (MIS), based on a flexible multispectral structured illumination probe (1.9 mm diameter) (Clancy et al. in Biomed Opt Express 2(11):3119-3128, 2011. doi: 10.1364/BOE.2.003119 ). This paper reports further hardware and analytical developments to improve the light pattern decoding result and increase the reconstruction accuracy. METHODS: The feasibility of using an 8-band multispectral camera with higher pattern-colour discrimination ability than normal RGB camera in this system was studied. Additionally, the "normalised cut" algorithm was investigated to improve pattern segmentation. RESULTS: The whole SL system was evaluated by phantom and in vivo experiments. Higher pattern identification performance than that of an RGB camera was recorded by using the multispectral camera (average precision >97%, average sensitivity >62%). An average of [Formula: see text] reconstruction error was achieved using the proposed pattern decoding method on a heart phantom at a working distance of approximately 10 cm. CONCLUSIONS: The experiment showed the superiority of the multispectral camera over the RGB camera in the spot identification step. The proposed pattern decoding algorithm underwent evaluations using different experiments, showing that it provided promising reconstruction results. The potential of using this system in MIS environments has been demonstrated.
Assuntos
Endoscopia/métodos , Interpretação de Imagem Assistida por Computador/métodos , Iluminação/métodos , Algoritmos , Estudos de Viabilidade , Humanos , Imagens de Fantasmas , Sensibilidade e EspecificidadeRESUMO
Huntington's disease (HD) is a genetic and neurodegenerative disease, leading to motor and cognitive dysfunction in HD patients. At cellular level, this disease is caused by the accumulation of mutant huntingtin (HTT) in different cells, and finally results in the dysfunction of different cells. To clean these mutant proteins, ubiquitin-proteasome system (UPS) and autophagy system are two critical pathways in the brain; however, little is known in other peripheral tissues. As mutant HTT affects different tissues progressively and might influence the UPS and autophagy pathways at early stages, we attempted to examine two clearance systems in HD models before the onset. Here, in vitro results showed that the accumulation of UPS signals with time was observed obviously in neuroblastoma and kidney cells, not in other cells. In HD transgenic mice, we observed the impairment of UPS, but not autophagy, over time in the cortex and striatum. In heart and muscle tissues, disturbance of autophagy was observed, whereas dysfunction of UPS was displayed in liver and lung. These results suggest that two protein clearance pathways are disturbed differentially in different tissues before the onset of HD, and enhancement of protein clearance at early stages might provide a potential stratagem to alleviate the progression of HD.
Assuntos
Doença de Huntington/genética , Doença de Huntington/patologia , Complexo de Endopeptidases do Proteassoma/genética , Ubiquitina/metabolismo , Animais , Autofagia/genética , Encéfalo/metabolismo , Encéfalo/patologia , Linhagem Celular , Modelos Animais de Doenças , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Proteína Huntingtina , Rim/metabolismo , Rim/patologia , Fígado/metabolismo , Fígado/patologia , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Transgênicos , Músculos/metabolismo , Músculos/patologia , Mutação/genética , Miocárdio/metabolismo , Miocárdio/patologia , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neuroblastoma/patologia , Ubiquitina/genéticaRESUMO
Dendritic cells (DCs) play an important role in initiating antitumour immune response. Tumour progression usually induces defects in DC maturation and thus tumour-bearing hosts exhibit immunosuppression and tumour escape. The previous studies showed that an exopolysaccharide (EPS) from a fungus, one anamorph of Cordyceps sinensis, inhibited tumour growth via activating immune response in the hosts. In view of the crucial actions of DCs in antitumour immunity, the present study aims to explore the effects of EPS on murine DCs. Murine DCs were derived from the bone marrow of C57BL/6 mice, and the effects of EPS on phenotype molecules and ingestion function of DCs were assayed using flow cytometry. Cytokine expressions of DCs were assayed by reverse transcriptase-PCR. Additionally, the level of phosphorylated signal transducers and activators of transcription 3 (p-STAT3) of DCs was evaluated using Western blotting. The results showed that EPS promoted the levels of surface molecules MHC II, CD40, CD80 and CD86 of DCs and decreased their ingestion ability. The mRNA expressions of cytokines (IL-12p40 and TNF-α) and inducible nitric oxide synthase were up-regulated by EPS. We also found that EPS significantly down-regulated p-STAT3 level of DCs. The results suggested that the promotion of DC's maturation and activation by EPS is probably related to the inhibition of STAT3 phosphorylation.
Assuntos
Cordyceps/imunologia , Células Dendríticas/imunologia , Polissacarídeos/farmacologia , Animais , Antígeno B7-2/imunologia , Antígeno B7-2/metabolismo , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Antígenos CD40/imunologia , Antígenos CD40/metabolismo , Diferenciação Celular , Cordyceps/química , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Endocitose , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica , Antígenos de Histocompatibilidade Classe II/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Subunidade p40 da Interleucina-12/imunologia , Subunidade p40 da Interleucina-12/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/imunologia , Óxido Nítrico Sintase Tipo II/metabolismo , Fenótipo , Polissacarídeos/imunologia , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Ativação Transcricional , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The immunogenicity of HLA-A*0201-restricted cytotoxic T lymphocyte (CTL) peptide in severe acute respiratory syndrome coronavirus (SARS-CoV) nuclear capsid (N) and spike (S) proteins was determined by testing the proteins' ability to elicit a specific cellular immune response after immunization of HLA-A2.1 transgenic mice and in vitro vaccination of HLA-A2.1 positive human peripheral blood mononuclearcytes (PBMCs). First, we screened SARS N and S amino acid sequences for allele-specific motif matching those in human HLA-A2.1 MHC-I molecules. From HLA peptide binding predictions (http://thr.cit.nih.gov/molbio/hla_bind/), ten each potential N- and S-specific HLA-A2.1-binding peptides were synthesized. The high affinity HLA-A2.1 peptides were validated by T2-cell stabilization assays, with immunogenicity assays revealing peptides N223-231, N227-235, and N317-325 to be the first identified HLA-A*0201-restricted CTL epitopes of SARS-CoV N protein. In addition, previous reports identified three HLA-A*0201-restricted CTL epitopes of S protein (S978-986, S1203-1211, and S1167-1175), here we found two novel peptides S787-795 and S1042-1050 as S-specific CTL epitopes. Moreover, our identified N317-325 and S1042-1050 CTL epitopes could induce recall responses when IFN-gamma stimulation of blood CD8+ T-cells revealed significant difference between normal healthy donors and SARS-recovered patients after those PBMCs were in vitro vaccinated with their cognate antigen. Our results would provide a new insight into the development of therapeutic vaccine in SARS.