RESUMO
Mimosa pudica L. (MP) is well-known plant in traditional medicinal system, especially in India. Unfortunately, leaves of MP are less explored. To determine the food and nutritional value of the neglected part of Mimosa pudica L. (MP), that is MP leaves, phytochemicals and metal ions of MP were quantified by newly developed HPLC and ICPOES-based methods. The content of phytochemicals observed using HPLC analysis for chlorogenic acid, catechin, and epicatechin was 141.823 (±8.171), 666.621 (±11.432), and 293.175 (±12.743) µg/g, respectively. Using GC/MS/MS analysis, fatty acid like oleic acid were identified. In ICP-OES analysis, a significant content of Na, K, Ca, Cu, Fe, Mg, Mn, and Zn was observed. The observed TPC and TFC for MP leaf extracts was 44.327 (±1.041) mg GAE/ g of wt. and 214.217 (±4.372) mg QCE/ g of wt., respectively. The DPPH assay depicted a strong antioxidant activity of MP leaf extracts with IC50 values of 0.796 (±0.081) mg/mL and a TEAC value of 0.0356 (±0.0003). A significant antacid activity (666â mg MP+400â mg CaCO3 >400â mg CaCO3 â«666â mg Gelusil) of MP leaves was noticed. The methanolic extract of MP leaves demonstrated anti-microbial activity against Staphylococcus aureus (15±2mm), Pseudomonas aeruginosa (12±2mm) and Escherichia coli (10±2mm). In silico studies confirmed the inâ vitro results obtained for antioxidant, antiacid, and anti-microbial activities. In addition, in silico studies revealed the anti-cancerous and anti-inflammatory potential of the MP leaves. In summary, this study demonstrated the medicinal significance of MP leaves and the conversion of agro-waste or the under-utilized part of MP into pharmaceutical potent materials. Consequently, the present study highlighted that MP leaves alone have medicinal importance with good nutritional utility and possess large promise in the pharma industry along with improving bio-valorization and the environment.
RESUMO
Today, due to the widespread outbreak of the deadly coronavirus, popularly known as COVID-19, the traditional classroom education has been shifted to computer-based learning. Students of various cognitive and psychological abilities participate in the learning process. However, most students are hesitant to provide regular and honest feedback on the comprehensiveness of the course, making it difficult for the instructor to ensure that all students are grasping the information at the same rate. The students' understanding of the course and their emotional engagement, as indicated via facial expressions, are intertwined. This paper attempts to present a three-dimensional DenseNet self-attention neural network (DenseAttNet) used to identify and evaluate student participation in modern and traditional educational programs. With the Dataset for Affective States in E-Environments (DAiSEE), the proposed DenseAttNet model outperformed all other existing methods, achieving baseline accuracy of 63.59% for engagement classification and 54.27% for boredom classification, respectively. Besides, DenseAttNet trained on all four multi-labels, namely boredom, engagement, confusion, and frustration has registered an accuracy of 81.17%, 94.85%, 90.96%, and 95.85%, respectively. In addition, we performed a regression experiment on DAiSEE and obtained the lowest Mean Square Error (MSE) value of 0.0347. Finally, the proposed approach achieves a competitive MSE of 0.0877 when validated on the Emotion Recognition in the Wild Engagement Prediction (EmotiW-EP) dataset.
RESUMO
Minichoromosome maintenance (MCM) proteins play key role in cell cycle progression by licensing DNA replication only once per cell cycle. These proteins are found to be overexpressed in cervical cancer cells. In this study, we depleted MCM4, one of the MCM 2-7 complex components by RNA interference (RNAi) in four cervical cancer cell lines. The four cell lines were selected on the basis of their human papillomavirus (HPV) infection: HPV16-positive SiHa, HPV18-positive ME-180, HPV16- and HPV18-positive CaSki, and HPV-negative C-33A. The MCM4-deficient cells irrespective of their HPV status grow for several generations and maintain regular cell cycle. We did not find any evidence of augmented response to a short-term (48 h) cisplatin treatment in these MCM4-deficient cells. However, MCM4-/HPV16+ SiHa cells cannot withstand a prolonged treatment (up to 5 days) of even a sublethal dosage of cisplatin. They show increased chromosomal instability compared to their control counterparts. On the other hand, MCM4-deficient CaSki cells (both HPV16+ and 18+) remain resistant to a prolonged exposure to cisplatin. Our study indicates that cervical cancer cells may be using excess MCMs as a backup for replicative stress; however, its regulatory mechanism is dependent on the HPV status of the cells.
Assuntos
Cisplatino/administração & dosagem , Resistencia a Medicamentos Antineoplásicos/genética , Componente 4 do Complexo de Manutenção de Minicromossomo/genética , Neoplasias do Colo do Útero/genética , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Feminino , Papillomavirus Humano 16/efeitos dos fármacos , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/efeitos dos fármacos , Papillomavirus Humano 18/patogenicidade , Humanos , Componente 4 do Complexo de Manutenção de Minicromossomo/antagonistas & inibidores , Interferência de RNA , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/virologiaRESUMO
Redox responsive biodegradable polymersomes comprising of poly(ethylene glycol)-polylactic acid-poly(ethylene glycol) [PEG-s-s-PLA-s-s-PLA-s-s-PEG] triblock copolymer with multiple disulfide linkages were developed to improve intracellular delivery and to enhance chemotherapeutic efficacy of doxorubicin in breast cancer with minimal cardiotoxicity. Folic acid and trastuzumab functionalized monodispersed polymersomes of size â¼150 nm were prepared by nanoprecipitation method while achieving enhanced doxorubicin loading of â¼32% in the polymersomes. Multiple redox responsive disulfide linkages were incorporated in the polymer in order to achieve complete disintegration of polymersomes in redox rich environment of cancer cells resulting in enhanced doxorubicin release as observed in in vitro release studies, where â¼90% doxorubicin release was achieved in pH 5.0 in the presence of 10 mM glutathione (GSH) as compared to â¼20% drug release in pH 7.4. Folic acid and trastuzumab mediated active targeting resulted in improved cellular uptake and enhanced apoptosis in in vitro studies in breast cancer cell lines. In vivo studies in Ehrlich ascites tumor bearing Swiss albino mice showed enhanced antitumor efficacy and minimal cardiotoxicity of polymersomes with â¼90% tumor regression as compared to â¼38% tumor regression observed with free doxorubicin. The results highlight therapeutic potential of the polymersomes as doxorubicin delivery nanocarrier in breast cancer therapy with its superior antitumor efficacy and minimal cardiotoxicity.
Assuntos
Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Portadores de Fármacos/química , Ácido Fólico/farmacologia , Neoplasias Mamárias Experimentais/tratamento farmacológico , Trastuzumab/farmacologia , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Doxorrubicina/farmacocinética , Doxorrubicina/uso terapêutico , Portadores de Fármacos/efeitos adversos , Portadores de Fármacos/síntese química , Liberação Controlada de Fármacos , Feminino , Humanos , Células MCF-7 , Camundongos , Oxirredução , Polietilenoglicóis/químicaRESUMO
Gallbladder cancer (GBC) is a rare but very aggressive most common digestive tract cancer with a high mortality rate due to delayed diagnosis at the advanced stage. Moreover, GBC progression shows asymptomatic characteristics making it impossible to detect at an early stage. In these circumstances, conventional therapy like surgery, chemotherapy, and radiotherapy becomes refractive. However, few studies reported some molecular markers like KRAS (Kirsten Rat Sarcoma) mutation, upregulation of HER2/neu, EGFR (Epidermal Growth Factor Receptor), and microRNAs in GBC. However, the absence of some specific early diagnostic and prognostic markers is the biggest hurdle for the therapy of GBC to date. The present study has been designed to identify some specific molecular markers for precise diagnosis, and prognosis, for successful treatment of the GBC. By In Silico a network-centric analysis of two microarray datasets; (GSE202479) and (GSE13222) from the Gene Expression Omnibus (GEO) database, shows 50 differentially expressed genes (DEGs) associated with GBC. Further network analysis revealed that 12 genes are highly interconnected based on the highest MCODE (Molecular Complex Detection) value, among all three genes; TRIP13 (Thyroid Receptor Interacting Protein), NEK2 (Never in Mitosis gene-A related Kinase 2), and TPX2 (Targeting Protein for Xklp2) having highest network interaction with transcription factors and miRNA suggesting critically associated with GBC. Further survival analysis data corroborate the association of these genes; TRIP13, NEK2, and TPX2 with GBC. Thus, TRIP13, NEK2, and TPX2 genes are significantly correlated with a greater risk of mortality, transforming them from mere biomarkers of the GBC for early detections and may emerge as prognostic markers for treatment.
Assuntos
Biomarcadores Tumorais , Neoplasias da Vesícula Biliar , Regulação Neoplásica da Expressão Gênica , Neoplasias da Vesícula Biliar/genética , Neoplasias da Vesícula Biliar/patologia , Neoplasias da Vesícula Biliar/metabolismo , Humanos , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Quinases Relacionadas a NIMA/genética , Quinases Relacionadas a NIMA/metabolismo , Simulação por Computador , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Redes Reguladoras de Genes , Perfilação da Expressão Gênica , Prognóstico , Carcinogênese/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismoRESUMO
Agro-waste is the outcome of the under-utilization of bioresources and a lack of knowledge to re-use this waste in proper ways or a circular economy approach. In the Indian medicinal system, the root of Cyperus scariosus (CS) is used at a large scale due to their vital medicinal properties. Unfortunately, the aerial part of CS is treated as agro-waste and is an under-utilized bioresource. Due to a lack of knowledge, CS is treated as a weed. This present study is the first ever attempt to explore CS leaves as medicinally and a nutrient rich source. To determine the food and nutritional values of the neglected part of Cyperus scariosus R.Br. (CS), i.e. CS leaves, phytochemicals and metal ions of CS were quantified by newly developed HPLC and ICPOES-based methods. The content of the phytochemicals observed in HPLC analysis for caffeic acid, catechin, epicatechin, trans-p-coumaric acid, and trans-ferulic acid was 10.51, 276.15, 279.09, 70.53, and 36.83 µg/g, respectively. In GC-MS/MS analysis, fatty acids including linolenic acid, phytol, palmitic acid, etc. were identified. In ICPOES analysis, the significant content of Na, K, Ca, Cu, Fe, Mg, Mn, and Zn was observed. The TPC and TFC of the CS leaves was 17.933 mg GAE eq./g and 130.767 mg QCE eq./g along with an IC50 value of 2.78 mg/mL in the DPPH assay and better antacid activity was measured than the standard (CaCO3). The methanolic extract of CS leaves showed anti-microbial activity against Staphylococcus aureus (15 ± 2 mm), Pseudomonas aeruginosa (12 ± 2 mm) and Escherichia coli (10 ± 2 mm). In silico studies confirmed the in vitro results obtained from the antioxidant, antiacid, and anti-microbial studies. In addition, in silico studies revealed the anti-cancerous and anti-inflammatory potential of the CS leaves. This study, thus, demonstrated the medicinal significance of the under-utilized part of CS and the conversion of agro-waste into mankind activity as a pharmaceutical potent material. Consequently, the present study highlighted that CS leaves have medicinal importance with good nutritional utility and have a large potential in the pharmaceutical industry along with improving bio-valorization and the environment.
Assuntos
Cyperus , Extratos Vegetais/química , Espectrometria de Massas em Tandem , Antioxidantes/análise , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/análise , Folhas de Planta/químicaRESUMO
Glycine betaine (GB) is an important osmolyte synthesized in response to different abiotic stresses, including salinity. The two known pathways of GB synthesis involve: 1) two step oxidation of choline (choline â betaine aldehyde â GB), generally found in plants, microbes and animals; and 2) three step methylation of glycine (glycine â sarcosine â dimethylglycine â GB), mainly found in halophilic archaea, sulphur bacteria and the cyanobacterium Aphanothece (Ap.) halophytica. Here, we transformed a salt-sensitive freshwater diazotrophic filamentous cyanobacterium Anabaena (An.) doliolum with N-methyltransferase genes (ApGSMT-DMT) from Ap. halophytica using the triparental conjugation method. The transformed An. doliolum synthesized and accumulated GB in cells, and showed increased salt tolerance and protection to nitrogenase activity. The salt responsiveness of the transformant was also apparent as GB synthesis increased with increasing concentrations of NaCl in the nutrient solution, and maximal [12.92 µmol (g dry weight)(-1)] in cells growing at 0.5 M NaCl. Therefore, the transformed cyanobacterium has changed its behaviour from preferring freshwater to halophily. This study may have important biotechnological implications for the development of stress tolerant nitrogen-fixing cyanobacteria as biofertilizers for sustainable agriculture.
Assuntos
Cianobactérias/enzimologia , Cianobactérias/fisiologia , Nitrogenase/metabolismo , Proteínas Metiltransferases/metabolismo , Tolerância ao Sal , Cloreto de Sódio/metabolismo , Cianobactérias/genética , Água Doce/microbiologia , Proteínas Metiltransferases/genética , Transformação BacterianaRESUMO
Breast cancer is the most prevalent cancer with the maximum number of cases worldwide. Early diagnosis of the cancer is necessary for an effective treatment plan. Due to a lack of awareness, diagnosis of breast cancer at an early stage is difficult. The present study aims to evaluate and compare the haematological and biochemical profiles of the early and late-stage breast cancer patient's data records. A retrospective cohort study was conducted on 56 breast cancer patients at the Institute of Medical Sciences, Banaras Hindu University India. Patient data records were obtained and haematological and biochemical parameters were arranged on an Excel sheet and analyzed. Random blood sugar (RBS), alkaline phosphates (ALP) levels, and urea levels were significantly high in patients with late-stage breast cancer (Tumor stage III and IV). At the advanced stage of breast cancer hemoglobin level falls and patients became anemic. Further large-scale studies with a greater number of patient data can help establish these parameters individually or in combination as prognostic and diagnostic markers in breast cancer staging.
RESUMO
Bowel cancer is a multifactorial disease arising from a combination of genetic predisposition and environmental factors. Detection of bowel cancer and its precursor lesions is predominantly performed by either visual inspection of the colonic mucosa during endoscopy or cross-sectional imaging. Most cases are diagnosed when the cancer is already at an advanced stage. These modalities are less reliable for detecting lesions at the earliest stages, when they are typically small or flat. Removal of lesions at the earliest possible stage reduces the risk of cancer death, which is largely due to a reduced risk of subsequent metastasis. In this review, we summarised the origin of bowel cancer and the mechanism of its metastasis. In particular, we reviewed a broad spectrum of literatures covering the biomechanics of bowel cancer and its measurement techniques that are pertinent to the successful development of a bowel cancer diagnostic device. We also reviewed existing bowel cancer diagnostic techniques that are available for clinical use. Finally, we outlined current clinical needs and highlighted the potential roles of medical robotics on early bowel cancer diagnosis.
Assuntos
Neoplasias Colorretais , Humanos , Neoplasias Colorretais/diagnóstico por imagem , Neoplasias Colorretais/patologia , Neoplasias Colorretais/cirurgia , Metástase NeoplásicaRESUMO
BACKGROUND: Masticatory forces generate various degrees of stress and strain in the periodontium of teeth which determine the clinical functions and load-bearing capacity of the teeth. There are few in vitro studies that have analyzed stress generated due to combined forces acting on the teeth. Thus, the objective of the present study was to do a comparative analysis of the influence of various stresses on the periodontal ligament and alveolar bone of maxillary central incisor with normal bone height and reduced bone height under simulated standard masticatory using finite element stress analysis. METHODOLOGY: A 3D model of the tooth was obtained with the help of ANSYS software. These models were subjected to various oblique forces, i.e., 100N and 235.9N, applied at 45° angle on the lingual surface of the maxillary central incisor and stress values were recorded in three dimensions. The results from FE analysis were analyzed using 3D Von Mises Criteria. RESULTS: It was observed that in healthy periodontium; it was observed that among the periodontal structure studied, the maximum stress levels were exerted on root followed by cortical bone, cancellous bone, and PDL, irrespective of the force, as compared to the diseased periodontium, in which the bone height was reduced, the maximum stresses were on root followed by cortical bone, PDL, and cancellous bone. CONCLUSION: The main factor governing the success of any periodontal procedure depends on the height of the remaining bone and the amount of force exerted on to the tooth and the stress generated within the tooth. The finite element method could be of substantial importance in this respect as it can assess the stresses of various occlusal forces on the periodontal ligament, root, cortical bone, and cancellous bone of teeth in a periodontally healthy and diseased state.
RESUMO
SARS-CoV2 pandemic exposed the limitations of artificial intelligence based medical imaging systems. Earlier in the pandemic, the absence of sufficient training data prevented effective deep learning (DL) solutions for the diagnosis of COVID-19 based on X-Ray data. Here, addressing the lacunae in existing literature and algorithms with the paucity of initial training data; we describe CovBaseAI, an explainable tool using an ensemble of three DL models and an expert decision system (EDS) for COVID-Pneumonia diagnosis, trained entirely on pre-COVID-19 datasets. The performance and explainability of CovBaseAI was primarily validated on two independent datasets. Firstly, 1401 randomly selected CxR from an Indian quarantine center to assess effectiveness in excluding radiological COVID-Pneumonia requiring higher care. Second, curated dataset; 434 RT-PCR positive cases and 471 non-COVID/Normal historical scans, to assess performance in advanced medical settings. CovBaseAI had an accuracy of 87% with a negative predictive value of 98% in the quarantine-center data. However, sensitivity was 0.66-0.90 taking RT-PCR/radiologist opinion as ground truth. This work provides new insights on the usage of EDS with DL methods and the ability of algorithms to confidently predict COVID-Pneumonia while reinforcing the established learning; that benchmarking based on RT-PCR may not serve as reliable ground truth in radiological diagnosis. Such tools can pave the path for multi-modal high throughput detection of COVID-Pneumonia in screening and referral.
Assuntos
COVID-19/complicações , Aprendizado Profundo , Sistemas Inteligentes , Processamento de Imagem Assistida por Computador/métodos , Pneumonia/diagnóstico , Radiografia Torácica/métodos , Tomografia Computadorizada por Raios X/métodos , Algoritmos , COVID-19/virologia , Humanos , Incidência , Índia/epidemiologia , Redes Neurais de Computação , Pneumonia/diagnóstico por imagem , Pneumonia/epidemiologia , Pneumonia/virologia , Estudos Retrospectivos , SARS-CoV-2/isolamento & purificaçãoRESUMO
PARP1 trapping at DNA lesion by pharmacological inhibitors has been exploited in several cancers exhibiting defects in DNA repair mechanisms. PARP1 hyperactivation is involved in therapeutic resistance in multiple cancers. The role of PARP1 in cervical cancer (CC) resistance and implication of PARP inhibitor is yet to be elucidated. Our data demonstrates significantly higher expression of PARP1 in primary cervical tumors and CC cell lines SiHa and ME180. Upon cisplatin treatment CC cells display significant overexpression of PARP1 and its hyperactivation. PARP inhibitor olaparib shows significant anti-proliferative effect on CC cells and drive loss of clonogenic survival and enhanced cell death in combination with cisplatin. PARP inhibited cells show delay in resolution of γH2A.X foci and prolonged late S and G2-M phase arrest resulting in apoptosis. Further, PARP inhibition disrupts the localization of base excision repair (BER) effector XRCC1 and non-homologous end joining (NHEJ) proteins Ku80 and XRCC4. Due to disrupted relocation of repair factors, cisplatin induced stalled replication forks collapse and convert into double strand breaks (DSBs). Interestingly, PARP inhibition also shows anti-migratory and anti-invasive properties in CC cells, increases anchorage independent cell death and induces anoikis. Collectively, our data demonstrates therapeutic potential of PARP inhibitor in cervical cancer.
Assuntos
Reparo do DNA/efeitos dos fármacos , Ftalazinas/farmacologia , Piperazinas/farmacologia , Neoplasias do Colo do Útero/patologia , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Cisplatino/farmacologia , Quebras de DNA de Cadeia Dupla/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Feminino , Inativação Gênica/efeitos dos fármacos , Humanos , Metástase Neoplásica , Proteínas Nucleares/metabolismo , Poli(ADP-Ribose) Polimerase-1/metabolismo , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologiaRESUMO
Context ⢠Isha Hatha yoga is a form of yoga practice that practitioners claim can improve health. Gaining a scientific understanding of its effects on health is a crucial step in claiming health benefits. Objective ⢠The study intended to investigate the effects of Isha Hatha yoga on core stability and standing balance in healthy volunteers. Design ⢠The research team designed a quasiexperimental study. Setting ⢠The setting was at Isha Yoga Center at Isha Foundation in Coimbatore, India. Participants ⢠Participants were individuals registered for a residential program providing training in Isha Hatha yoga at the Isha Yoga Center. Intervention ⢠The training consisted of 21 d of training in Isha Hatha yoga, including Upa yoga, Surya Kriya, Angamardhana, Bhuta Suddhi, and asanas. Outcome Measures ⢠The single-leg stork test and the plank test were conducted before and after the training. Results ⢠Significant increases occurred in the standing-stork test scores on the right (P = .014) and left (P = .033) sides and in the plank test scores (P < .001). Conclusions ⢠The current study concluded that healthy volunteers who underwent 21 d of training in Isha Hatha yoga showed significant improvements in core strength and balance.
Assuntos
Músculo Esquelético/fisiologia , Equilíbrio Postural/fisiologia , Yoga , Adulto , Humanos , Resultado do TratamentoRESUMO
Cell cycle regulators cyclin D1 and cyclin E2 function in G1/S transition by activating downstream cyclin-dependent kinases. Deregulated expression of these cyclins has been reported in various cancers. However, little is known about their clinical significance in gastric carcinoma. We aimed to explore that whether there is differential expression of these cyclins in clinically distinct gastric cancer patients. In this study we recruited a total of 92 subjects including 20 controls and 72 cases of histopathologically proven gastric carcinoma. Expression profiling at transcript level was done by semiquantitative RT-PCR and of protein by immunohistochemistry. Receiver operator characteristics analysis was done for determining diagnostic utility of cyclin D1 and cyclin E2. We demonstrate that cyclins D1 and E2 are frequently overexpressed in early stages of gastric carcinoma. Interestingly, expression of cyclins D1 and E2 significantly correlates with different clinical parameters such as gender, histological type (intestinal and diffuse), tumor location (proximal, middle, and distal), tumor differentiation (differentiated and undifferentiated), tumor invasion (serosal, lymphatic, and venous) and tumor metastasis (lymph node, peritoneal, ascites, and liver). Cyclin D1 has significantly higher sensitivity and specificity as diagnostic biomarker than cyclin E2. Our results suggest that overexpression of cyclin D1 and cyclin E2 is an early event in gastric carcinogenesis. The differential expression of these cyclins may be useful as diagnostic biomarkers for early detection of gastric carcinoma.
Assuntos
Ciclina D1/metabolismo , Ciclinas/metabolismo , Neoplasias Gástricas/metabolismo , Biomarcadores Tumorais/metabolismo , Estudos de Casos e Controles , Ciclina D1/genética , Ciclinas/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Curva ROC , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologiaRESUMO
CXCL12 is a small pro-inflammatory chemo-attractant cytokine which signals through chemokine receptor CXCR4. The importance of CXCL12/CXCR4 axis is coming to the fore in several divergent signaling pathway-initiating signals related to cell survival and/or proliferation and cancer metastasis. In the present study we have investigated whether deregulation in CXCR4 signaling (as a consequence of deregulated expression of CXCL12) modulate the metastatic potential of cervical carcinoma cells. We demonstrate that CXCL12 is frequently down regulated and its promoter is hypermethylated in cervical cancer cell lines and primary tumor biopsies. Exogenous treatment of cervical cancer cell lines (HeLa, SiHa and C-33A) with recombinant CXCL12 inhibited the metastasis promoting cell migration, cell invasion and anchorage independent cell growth events. Although this study will need further in vivo validation, our observations suggest that (a) silencing of CXCL12 in cervical cancer cells may be critical in migration and invasion, the key events in cancer cell metastases; (b) cervical cancer cells having down regulated CXCL12 are more prone to being attracted to CXCL12 expressed at secondary sites of metastases; and (c) CXCL12 inhibits anchorage independent cell growth via anoikis. These findings suggest the tumor suppressor functions of CXCL12 in cervical cancer.
Assuntos
Quimiocina CXCL12/genética , Metástase Neoplásica , Microambiente Tumoral/genética , Neoplasias do Colo do Útero/genética , Apoptose/genética , Movimento Celular/genética , Proliferação de Células/genética , Quimiocina CXCL12/biossíntese , Feminino , Regulação Neoplásica da Expressão Gênica , Células HeLa , Humanos , Invasividade Neoplásica/genética , Neoplasias do Colo do Útero/patologiaRESUMO
Adenomatoid odontogenic tumour is a benign (hamartomatous), noninvasive lesion with slow but progressive growth. The 3 variants--follicular, extrafollicular and peripheral--present with identical histological findings. This report describes a patient with a large adenomatoid odontogenic tumour in the mandible, with the involved mandibular canine being pushed to the contralateral side. The paper also provides a refresher for general dental practitioners about various diagnostic aspects of this tumour and highlights the controversies regarding its origin and management in light of recent findings.
Assuntos
Neoplasias Mandibulares/patologia , Tumores Odontogênicos/patologia , Adolescente , Diagnóstico Diferencial , Feminino , HumanosRESUMO
AIM: The fundamental events for cancer progression and metastases include loss of cell adhesion, cell proliferation, anchorage-independent cell growth (evading anoikis), cell migration and cell invasion. All these events leading to cancer progression happen in a favorable nurturing tumor microenvironment. This study was designed to explore the anti-tumor activity of staurosporine (a nonspecific protein kinase inhibitor) in the tumor microenvironment of cervical cancer. MAIN METHODS: The anti-tumor activity of staurosporine was investigated by cell adhesion assay, colony formation assay, apoptosis assay and quantitative real-time polymerase chain reaction (PCR) in cervical cancer cell lines. KEY FINDINGS: The cell adhesion assay showed that staurosporine induces adhesion of cervical cancer cells to the extracellular matrix (ECM) protein fibronectin. The soft agar colony formation assay showed that staurosporine inhibits both the number and size of colony formation in a dose dependent manner and also induces adherent tendency in the cancer cells. Staurosporine also induces prominent apoptosis in single cell suspensions compared to adherent cells. Stroma cell induced transcription of matrix metalloprotease 1 (MMP1) and matrix metalloprotease 2 (MMP2) in cervical cancer cells was inhibited by staurosporine. SIGNIFICANCE: Our results indicate that staurosporine induces anti-tumor response in the cervical tumor microenvironment by inhibiting the fundamental events for cancer progression and metastases. The present study represents an attractive area for further research and opens up new avenues towards the understanding of cervical cancer therapeutics.
Assuntos
Antineoplásicos/farmacologia , Colo do Útero/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Estaurosporina/farmacologia , Microambiente Tumoral/efeitos dos fármacos , Neoplasias do Colo do Útero/tratamento farmacológico , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Colo do Útero/metabolismo , Colo do Útero/patologia , Regulação para Baixo/efeitos dos fármacos , Feminino , Humanos , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 2 da Matriz/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: The cyclin-dependent kinase inhibitor p27(Kip1) is known to act as a putative tumor suppressor in several human cancers, including cervical cancer. Down-regulation of p27(Kip1) may occur either through transcription inhibition or through phosphorylation-dependent proteolytic degradation. As yet, the mechanism underlying p27(Kip1) down-regulation and its putative downstream effects on cervical cancer development are poorly understood. Here we assessed the expression and sub-cellular localization of p27(Kip1) and its effects on proliferation, cell cycle progression and (inhibition of) apoptosis in cervical cancer cells. METHODS: Primary cervical cancer samples (n = 70), normal cervical tissue samples (n = 30) and cervical cancer-derived cell lines (n = 8) were used to assess the expression of p27(Kip1) and AKT1 by RT-PCR, Western blotting and immunohistochemistry, respectively. The effects of the PI3K inhibitor LY294004 and the proteasome inhibitor MG132 on cervical cancer cell proliferation were investigated using a MTT assay. Apoptosis and cell cycle analyses were carried out using flow cytometry, and sub-cellular p27(Kip1) localization analyses were carried out using immunofluorescence assays. RESULTS: We observed p27(Kip1) down-regulation (p = 0.045) and AKT1 up-regulation (p = 0.046) in both the primary cervical cancer samples and the cervical cancer-derived cell lines, compared to the normal cervical tissue samples tested. Treatment of cervical cancer-derived cell lines with the PI3K inhibitor LY294002 resulted in a reduced AKT1 activity. We also observed a dose-dependent inhibition of cell viability after treatment of these cell lines with the proteasome inhibitor MG132. Treatment of the cells with LY294002 resulted in a G1 cell cycle arrest, a nuclear expression of p27(Kip1), and a cytoplasmic p27(Kip1) accumulation after subsequent treatment with MG132. Additionally, we found that the synergistic effect of MG132 and LY294002 resulted in a sub-G1 cell cycle arrest and apoptosis induction through poly (ADP-ribose) polymerase (PARP) cleavage. CONCLUSION: Our data suggest that p27(Kip1) down-regulation in cervical cancer cells is primarily regulated through PI3K/AKT-mediated proteasomal degradation. The observed synergistic effect of the MG132 and LY294002 inhibitors may form a basis for the design of novel cervical cancer therapies.
Assuntos
Carcinoma de Células Escamosas/patologia , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias do Colo do Útero/patologia , Adulto , Apoptose/fisiologia , Western Blotting , Carcinoma de Células Escamosas/metabolismo , Pontos de Checagem do Ciclo Celular/fisiologia , Regulação para Baixo , Feminino , Citometria de Fluxo , Células HeLa , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Neoplasias do Colo do Útero/metabolismoRESUMO
UNLABELLED: The purpose of this study was to assess the feasibility of PET imaging of oncogene VPAC1 receptors overexpressed in human breast cancer cells. METHODS: Vasoactive intestinal peptide (VIP) analog (TP3982) was synthesized to harbor a carboxy-terminus lysine (Lys) residue separated from VIP-asparagine (Asn(28)) by 4-aminobutyric acid (Aba) as a spacer. Lys was derivatized with diaminopropionic acid coupled to a pair of dibenzoylthioglycolic acid residues as protecting groups. The analog was labeled with (64)Cu at pH 9 ((64)Cu-TP3982) and (99m)Tc at pH 12 ((99m)Tc-TP3982). (99m)Tc-TP3982 and VIP derivatized with Aba-GAGG and labeled with (99m)Tc ((99m)Tc-TP3654) were used as reference agents. Smooth muscle relaxivity assays performed with each derivative and compared with unaltered VIP(28) demonstrated functional integrity. In vitro stability of (64)Cu-TP3982 was determined by challenging the complex with 100-mol excess of diethylenetriaminepentaacetic acid (DTPA), human serum albumin (HSA), and cysteine. In vivo stability was determined in urine and serum for up to 24 h. The mass of the Cu-TP3982 complex was determined by mass spectrometry. Human T47D breast tumor xenografts were grown in athymic nude mice. Planar scintigraphic imaging was performed at 4 and 24 h after the intravenous administration of (99m)Tc-TP3982 and (99m)Tc-TP3654 and PET imaging was performed using a small animal MOSAIC PET scanner, also at 4 and 24 h after injection of (64)Cu-TP3982. Tissue-distribution studies were also performed. In a separate experiment, receptors were blocked by intravenous injection of authentic VIP(28) 30 min before the administration of (64)Cu-TP3982 and tissue distribution was examined. RESULTS: (64)Cu-TP3982 labeling yields were 98% +/- 1.2% and those for (99m)Tc-TP3982 and (99m)Tc-TP3654 were 98.2% +/- 1.1% and 97% +/- 1.6%, respectively. The biologic activity of both VIP analogs was uncompromised. When (64)Cu-TP3982 was challenged with 100-mol excess of DTPA, HSA, or cysteine, >98% radioactivity remained as (64)Cu-TP3982. In vivo, >98% of (64)Cu circulating in plasma remained as (64)Cu-TP3982. Of the (64)Cu excreted in urine 4, 20, and 24 h after injection, >98%, 89.9% +/- 0.9%, and 85% +/- 3%, respectively, were bound to TP3982. The mass of Cu-TP3982 as determined by surface-enhanced laser desorption/ionization time of flight (SELDI-TOF) was 4,049.7 Da. Four hours after receptor blocking with VIP(28), there was a significant reduction in uptake of all tissues except in the liver. With (64)Cu-TP3982, the 4-h postinjection tumor uptake was 10.8 +/- 2.1 %ID/g versus 0.5 +/- 0.02 %ID/g and 0.24 +/- 0.08 %ID/g for (99m)Tc-TP3982 and (99m)Tc-TP3654, respectively. Twenty-four hours after injection, the corresponding numbers were 17 +/- 0.7 %ID/g, 0.77 +/- 0.1 %ID/g, and 0.23 +/- 0.1 %ID/g. The severalfold greater uptake (21.2-74) of (64)Cu-TP3982 is attributable to the in vivo stability of the agent. CONCLUSION: The results suggest that the uncompromised biologic activity and the significantly greater tumor uptake of (64)Cu-TP3982, combined with the high sensitivity and enhanced resolution of PET imaging, make (64)Cu-TP3982 highly desirable for further studies in PET imaging of oncogene receptors overexpressed in breast and other types of cancers.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Radioisótopos de Cobre/farmacocinética , Peptídeos/farmacocinética , Receptores de Peptídeo Intestinal Vasoativo/metabolismo , Peptídeo Intestinal Vasoativo/farmacocinética , Animais , Biomarcadores Tumorais , Radioisótopos de Cobre/química , Radioisótopos de Cobre/farmacologia , Relação Dose-Resposta a Droga , Estabilidade de Medicamentos , Feminino , Contração Isométrica/efeitos dos fármacos , Masculino , Taxa de Depuração Metabólica , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Peso Molecular , Músculo Liso/efeitos dos fármacos , Oligopeptídeos/química , Oligopeptídeos/farmacocinética , Gambás , Especificidade de Órgãos , Peptídeos/química , Peptídeos/farmacologia , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacocinética , Compostos Radiofarmacêuticos/farmacologia , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tecnécio/química , Tecnécio/farmacocinética , Tecnécio/farmacologia , Distribuição Tecidual , Tomografia Computadorizada de Emissão/métodos , Peptídeo Intestinal Vasoativo/química , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
Frovatriptan, a 5-HT1B and 5-HT1D receptor agonist, is used for the treatment of acute migraine attack. This molecule is classified into second line therapy because of its slow onset of action (peak response obtained after 4 hours of administration) and low bioavailability (25%). Moreover, its therapy is the most costly among all triptans. Attempt has been made in present work to suggest a way out to fasten its onset of action and to enhance its bioavailability. Prepared tablets were evaluated by physicochemical tests, in vitro permeation studies, ex vivo permeation studies, and histopathological studies. Suitable mathematical calculations were performed to calculate the minimum amount of bioavailability that could be enhanced. Tablets containing chitosan (5% w/w) were found to give optimum results. Prepared tablets can double the bioavailability of frovatriptan and can initiate its response within 10 minutes of its administration. Suggestive alternative has the potential to increase the efficacy of frovatriptan for treating acute migraine attack.