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1.
Int J Mol Sci ; 23(22)2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36430568

RESUMO

Yersiniosis, caused by Yersinia enterocolitica, is the third most rampant zoonotic disease in Europe; the pathogen shows high antibiotic resistance. Herbs have multiple anti-microbial components that reduce microorganism resistance. Therefore, an extract of Picrorhiza kurroa (P. kurroa) was evaluated for potential antimicrobial activity. We report that the ethanolic extract of P. kurroa showed effective antimicrobial activity (zone of inhibition: 29.8 mm, Minimum inhibitory concentration (MIC): 2.45 mg/mL, minimum bactericidal concentration (MBC): 2.4 mg/mL) against Yersinia enterocolitica. Potential bioactive compounds from P. kurroa were identified using LC-MS, namely, cerberidol, annonidine A, benzyl formate, picroside-1, and furcatoside A. P. kurroa showed effective antimicrobial potential in skim milk at different pH, acidity, and water activity levels. P. kurroa affected the physiology of Yersinia enterocolitica and reduced the number of live cells. Yersinia enterocolitica, when incubated with P. kurroa extract, showed lower toxin production. Picroside-1 was isolated and showed higher antimicrobial potential in comparison to the standard antibiotic. Picroside-1 lysed the Yersinia enterocolitica cells, as observed under scanning electron microscopy. Docking revealed that picroside-1 (ligand) showed both hydrophilic and hydrophobic interactions with the dihydrofolate reductase (DHFR) protein of Yersinia enterocolitica and that DHFR is a possible drug target. The high activity and natural origin of Picroside-1 justify its potential as a possible drug candidate for Yersinia enterocolitica.


Assuntos
Anti-Infecciosos , Picrorhiza , Yersinia enterocolitica , Picrorhiza/química , Picrorhiza/metabolismo , Anti-Infecciosos/farmacologia , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/metabolismo
2.
Inorg Chem ; 59(22): 16168-16177, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33103424

RESUMO

Superbenzene porphyrin conjugates find wide range of applications from nonlinear optical materials to semiconductors. Herein, we report the synthesis and characterization of 5,15-bis(3,5-di-tert-butylphenyl)-10,20-bis(pentaphenylphenyl)phenylporphyrin and its Zinc-metallated complex. Oxidative planarization of 5,15-bis(3,5-di-tert-butylphenyl)-10,20-bis(pentaphenylphenyl)phenylporphyrin and its metallated complex was carried out by using NOBF4 as an oxidizing agent. The formation of superbenzene porphyrin conjugates validates its Scholl type reactions. The laboratory-synthesized porphyrin conjugates were characterized experimentally using spectroscopic techniques such as 1H NMR, 13C NMR, electron spin resonance, and ultraviolet-visible spectroscopy for structural conformation. In addition, density functional theory calculations were carried out to validate the experimental results. The theoretical and experimental results show that the 4-(pentaphenylphenyl)phenyl ligand increases the stability, optical properties, and rate of planarization of synthesized porphyrins. The conjugates exhibited intense and distant electronic communication between two hexabenzocoronene sites, taking advantage of porphyrin as a π-spacer. The π-radical cation has also been found to be an intermediate in oxidative C-C bond formation. NICS calculations support such a conclusion.

3.
J Nanosci Nanotechnol ; 17(4): 2525-530, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29652120

RESUMO

Guanidino-graphene has been synthesized by the reaction of bromoamine with reduced graphene oxide and characterized by FT-IR, Raman, TGA, powder XRD, TEM, SEM, and zeta potential. It is a cheap, heterogeneous and environmentally benign solid base catalyst used for cascade Aldol-Michael-oxidation in the synthesis of chalcone, flavonoids.

4.
Proc Natl Acad Sci U S A ; 111(40): E4224-33, 2014 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-25246575

RESUMO

Initiator tRNAs are special in their direct binding to the ribosomal P-site due to the hallmark occurrence of the three consecutive G-C base pairs (3GC pairs) in their anticodon stems. How the 3GC pairs function in this role, has remained unsolved. We show that mutations in either the mRNA or 16S rRNA leading to extended interaction between the Shine-Dalgarno (SD) and anti-SD sequences compensate for the vital need of the 3GC pairs in tRNA(fMet) for its function in Escherichia coli. In vivo, the 3GC mutant tRNA(fMet) occurred less abundantly in 70S ribosomes but normally on 30S subunits. However, the extended SD:anti-SD interaction increased its occurrence in 70S ribosomes. We propose that the 3GC pairs play a critical role in tRNA(fMet) retention in ribosome during the conformational changes that mark the transition of 30S preinitiation complex into elongation competent 70S complex. Furthermore, treating cells with kasugamycin, decreasing ribosome recycling factor (RRF) activity or increasing initiation factor 2 (IF2) levels enhanced initiation with the 3GC mutant tRNA(fMet), suggesting that the 70S mode of initiation is less dependent on the 3GC pairs in tRNA(fMet).


Assuntos
RNA Mensageiro/genética , RNA de Transferência de Metionina/genética , Ribossomos/metabolismo , Aminoglicosídeos/farmacologia , Anticódon/genética , Sequência de Bases , Sítios de Ligação/genética , Northern Blotting , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/metabolismo , Mutação , Iniciação Traducional da Cadeia Peptídica/genética , Fator de Iniciação 2 em Procariotos/metabolismo , RNA Mensageiro/metabolismo , RNA Ribossômico 16S/metabolismo , RNA de Transferência de Metionina/metabolismo , Proteínas Ribossômicas/metabolismo
5.
Nucleic Acids Res ; 41(9): 4963-75, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23530111

RESUMO

The accuracy of pairing of the anticodon of the initiator tRNA (tRNA(fMet)) and the initiation codon of an mRNA, in the ribosomal P-site, is crucial for determining the translational reading frame. However, a direct role of any ribosomal element(s) in scrutinizing this pairing is unknown. The P-site elements, m(2)G966 (methylated by RsmD), m(5)C967 (methylated by RsmB) and the C-terminal tail of the protein S9 lie in the vicinity of tRNA(fMet). We investigated the role of these elements in initiation from various codons, namely, AUG, GUG, UUG, CUG, AUA, AUU, AUC and ACG with tRNA(fMet(CAU) (tRNA(fMet) with CAU anticodon); CAC and CAU with tRNA(fMet(GUG); UAG with tRNA(fMet(CAU) ; UAC with tRNA(fMet(GUG) ; and AUC with tRNA(fMet(GUG) using in vivo and computational methods. Although RsmB deficiency did not impact initiation from most codons, RsmD deficiency increased initiation from AUA, CAC and CAU (2- to 3.6-fold). Deletion of the S9 C-terminal tail resulted in poorer initiation from UUG, GUG and CUG, but in increased initiation from CAC, CAU and UAC codons (up to 4-fold). Also, the S9 tail suppressed initiation with tRNA(fMet(CAU) lacking the 3GC base pairs in the anticodon stem. These observations suggest distinctive roles of 966/967 methylations and the S9 tail in initiation.


Assuntos
Proteínas de Escherichia coli/química , Escherichia coli/genética , Iniciação Traducional da Cadeia Peptídica , RNA Ribossômico 16S/química , RNA de Transferência de Metionina/química , Proteínas Ribossômicas/química , Ribossomos/química , Anticódon , Códon , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Metilação , Simulação de Dinâmica Molecular , Mutação , RNA Mensageiro/química , RNA Ribossômico 16S/metabolismo , RNA de Transferência de Metionina/genética , RNA de Transferência de Metionina/metabolismo , Proteína S9 Ribossômica , Proteínas Ribossômicas/genética , Deleção de Sequência
6.
Life Sci Alliance ; 7(8)2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38830771

RESUMO

Dengue fever, a neglected tropical arboviral disease, has emerged as a global health concern in the past decade. Necessitating a nuanced comprehension of the intricate dynamics of host-virus interactions influencing disease severity, we analysed transcriptomic patterns using bulk RNA-seq from 112 age- and gender-matched NS1 antigen-confirmed hospital-admitted dengue patients with varying severity. Severe cases exhibited reduced platelet count, increased lymphocytosis, and neutropenia, indicating a dysregulated immune response. Using bulk RNA-seq, our analysis revealed a minimal overlap between the differentially expressed gene and transcript isoform, with a distinct expression pattern across the disease severity. Severe patients showed enrichment in retained intron and nonsense-mediated decay transcript biotypes, suggesting altered splicing efficiency. Furthermore, an up-regulated programmed cell death, a haemolytic response, and an impaired interferon and antiviral response at the transcript level were observed. We also identified the potential involvement of the RBM39 gene among others in the innate immune response during dengue viral pathogenesis, warranting further investigation. These findings provide valuable insights into potential therapeutic targets, underscoring the importance of exploring transcriptomic landscapes between different disease sub-phenotypes in infectious diseases.


Assuntos
Processamento Alternativo , Vírus da Dengue , Dengue Grave , Humanos , Processamento Alternativo/genética , Feminino , Masculino , Vírus da Dengue/genética , Adulto , Dengue Grave/genética , Dengue Grave/imunologia , Dengue Grave/virologia , Pessoa de Meia-Idade , Transcriptoma/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Perfilação da Expressão Gênica/métodos , Imunidade Inata/genética , Dengue/genética , Dengue/imunologia , Dengue/virologia , Adulto Jovem , Índice de Gravidade de Doença , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia
7.
J Bacteriol ; 195(16): 3524-30, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23729652

RESUMO

The ribosomal P-site hosts the peptidyl-tRNAs during translation elongation. Which P-site elements support these tRNA species to maintain codon-anticodon interactions has remained unclear. We investigated the effects of P-site features of methylations of G966, C967, and the conserved C-terminal tail sequence of Ser, Lys, and Arg (SKR) of the S9 ribosomal protein in maintenance of the translational reading frame of an mRNA. We generated Escherichia coli strains deleted for the SKR sequence in S9 ribosomal protein, RsmB (which methylates C967), and RsmD (which methylates G966) and used them to translate LacZ from its +1 and -1 out-of-frame constructs. We show that the S9 SKR tail prevents both the +1 and -1 frameshifts and plays a general role in holding the P-site tRNA/peptidyl-tRNA in place. In contrast, the G966 and C967 methylations did not make a direct contribution to the maintenance of the translational frame of an mRNA. However, deletion of rsmB in the S9Δ3 background caused significantly increased -1 frameshifting at 37°C. Interestingly, the effects of the deficiency of C967 methylation were annulled when the E. coli strain was grown at 30°C, supporting its context-dependent role.


Assuntos
Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Elongação Traducional da Cadeia Peptídica/fisiologia , Sequência de Aminoácidos , Códon , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Mutação da Fase de Leitura , Metilação , Modelos Moleculares , Conformação Proteica , Ribossomos , Thermus thermophilus/genética , Thermus thermophilus/metabolismo
8.
Adv Healthc Mater ; 12(26): e2300842, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37402278

RESUMO

Circulating tumor cells (CTCs) are established as distinct cancer biomarkers for diagnosis, as preclinical models, and therapeutic targets. Their use as preclinical models is limited owing to low purity after isolation and the lack of effective techniques to create 3D cultures that accurately mimic in vivo conditions. Herein, a two-component system for detecting, isolating, and expanding CTCs to generate multicellular tumor spheroids that mimic the physiology and microenvironment of the diseased organ is proposed. First, an antifouling biointerface on magnetic beads is fabricated by adding a bioinert polymer layer and conjugation of biospecific ligands to isolate cancer cells, dramatically enhancing the selectivity and purity of the isolated cancer cells. Next, the isolated cells are encapsulated into self-degradable hydrogels synthesized using a thiol-click approach. The hydrogels are mechanochemically tuned to enable tumor spheroid growth to a size greater than 300 µm and to further release the grown spheroids while retaining their tumor-like characteristics. In addition, drug treatment highlights the need for 3D culture environments rather than conventional 2D culture. The designed biomedical matrix shows potential as a universal method to ensure mimicry of in vivo tumor characteristics in individual patients and to improve the predictability of preclinical screening of personalized therapeutics.


Assuntos
Células Neoplásicas Circulantes , Humanos , Avaliação Pré-Clínica de Medicamentos/métodos , Polímeros/farmacologia , Esferoides Celulares , Hidrogéis/farmacologia , Microambiente Tumoral
9.
Gels ; 9(12)2023 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-38131924

RESUMO

Hydrogel-based artificial scaffolds are essential for advancing cell culture models from 2D to 3D, enabling a more realistic representation of physiological conditions. These hydrogels can be customized through crosslinking to mimic the extracellular matrix. While the impact of extracellular matrix scaffolds on cell behavior is widely acknowledged, mechanosensing has become a crucial factor in regulating various cellular functions. cancer cells' malignant properties depend on mechanical cues from their microenvironment, including factors like stiffness, shear stress, and pressure. Developing hydrogels capable of modulating stiffness holds great promise for better understanding cell behavior under distinct mechanical stress stimuli. In this study, we aim to 3D culture various cancer cell lines, including MCF-7, HT-29, HeLa, A549, BT-474, and SK-BR-3. We utilize a non-degradable hydrogel formed from alpha acrylate-functionalized dendritic polyglycerol (dPG) and thiol-functionalized 4-arm polyethylene glycol (PEG) via the thiol-Michael click reaction. Due to its high multivalent hydroxy groups and bioinert ether backbone, dPG polymer was an excellent alternative as a crosslinking hub and is highly compatible with living microorganisms. The rheological viscoelasticity of the hydrogels is tailored to achieve a mechanical stiffness of approximately 1 kPa, suitable for cell growth. Cancer cells are in situ encapsulated within these 3D network hydrogels and cultured with cell media. The grown tumor spheroids were characterized by fluorescence and confocal microscopies. The average grown size of all tumoroid types was ca. 150 µm after 25 days of incubation. Besides, the stability of a swollen gel remains constant after 2 months at physiological conditions, highlighting the nondegradable potential. The successful formation of multicellular tumor spheroids (MCTSs) for all cancer cell types demonstrates the versatility of our hydrogel platform in 3D cell growth.

10.
Eur J Med Chem ; 252: 115299, 2023 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-36996716

RESUMO

Malaria is a tropical disease with significant morbidity and mortality burden caused by Plasmodium species in Africa, the Middle East, Asia, and South America. Pathogenic Plasmodium species have lately become increasingly resistant to approved chemotherapeutics and combination therapies. Therefore, there is an emergent need for identifying new druggable targets and novel chemical classes against the parasite. Falcipains, cysteine proteases required for heme metabolism in the erythrocytic stage, have emerged as promising drug targets against Plasmodium species that infect humans. This perspective discusses the biology, biochemistry, structural features, and genetics of falcipains. The efforts to identify selective or dual inhibitors and their structure-activity relationships are reviewed to give a perspective on the design of novel compounds targeting falcipains for antimalarial activity evaluating reasons for hits and misses for this important target.


Assuntos
Antimaláricos , Plasmodium , Humanos , Antimaláricos/química , Plasmodium falciparum , Inibidores de Cisteína Proteinase/farmacologia , Inibidores de Cisteína Proteinase/química , Relação Estrutura-Atividade
11.
ACS Sens ; 8(2): 443-464, 2023 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-36683281

RESUMO

Porphyrinoids and their analogous compounds play an important role in biosensing applications on account of their unique and versatile catalytic, coordination, photophysical, and electrochemical properties. Their remarkable arrays of properties can be finely tuned by synthetically modifying the porphyrinoid ring and varying the various structural parameters such as peripheral functionalization, metal coordination, and covalent or physical conjugation with other organic or inorganic scaffolds such as nanoparticles, metal-organic frameworks, and polymers. Porphyrinoids and their organic-inorganic conjugates are not only used as responsive materials but also utilized for the immobilization and embedding of biomolecules for applications in wearable devices, fast sensing devices, and other functional materials. The present review delineates the impact of different porphyrinoid conjugates on their physicochemical properties and their specificity as biosensors in a range of applications. The newest porphyrinoid types and their synthesis, modification, and functionalization are presented along with their advantages and performance improvements.


Assuntos
Técnicas Biossensoriais , Estruturas Metalorgânicas , Nanopartículas , Dispositivos Eletrônicos Vestíveis , Estruturas Metalorgânicas/química , Polímeros/química , Nanopartículas/química
12.
iScience ; 26(12): 108336, 2023 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-38025778

RESUMO

COVID-19 pandemic saw emergence of multiple SAR-CoV-2 variants. Exacerbated risk of severe outcome and hospital admissions led us to comprehend differential host-immune kinetics associated with SARS-CoV-2 variants. Longitudinal investigation was conducted through different time periods of Pre-VOC and VOCs (Delta & Omicron) mapping host transcriptome features. Robust antiviral type-1 interferon response marked Omicron infection, which was largely missing during Pre-VOC and Delta waves. SARS-CoV-2-host protein-protein interactions and docking complexes highlighted N protein to interact with HNRNPA1 in Pre-VOC, demonstrating its functional role for enhanced viral replication. Omicron revealed enhanced binding efficiency of LARP1 to N protein, probably potentiating antiviral effects of LARP1. Differential expression of zinc finger protein genes, especially in Omicron, mechanistically support induction of strong IFN (Interferon) response, thereby strengthening early viral clearance. Study highlights eventual adaptation of host to immune activation patterns that interrupt virus evolution with enhanced immune-evasion mutations and counteraction mechanisms, delimiting the next phase of COVID-19 pandemic.

13.
Curr Pharm Des ; 29(39): 3137-3153, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38031774

RESUMO

One-third of people will be diagnosed with cancer at some point in their lives, making it the second leading cause of death globally each year after cardiovascular disease. The complex anticancer molecular mechanisms have been understood clearly with the advent of improved genomic, proteomic, and bioinformatics. Our understanding of the complex interplay between numerous genes and regulatory genetic components within cells explaining how this might lead to malignant phenotypes has greatly expanded. It was discovered that epigenetic resistance and a lack of multitargeting drugs were highlighted as major barriers to cancer treatment, spurring the search for innovative anticancer treatments. It was discovered that epigenetic resistance and a lack of multitargeting drugs were highlighted as major barriers to cancer treatment, spurring the search for innovative anticancer treatments. Many popular anticancer drugs, including irinotecan, vincristine, etoposide, and paclitaxel, have botanical origins. Actinomycin D and mitomycin C come from bacteria, while bleomycin and curacin come from marine creatures. However, there is a lack of research evaluating the potential of algae-based anticancer treatments, especially in terms of their molecular mechanisms. Despite increasing interest in the former, and the promise of the compounds to treat tumours that have been resistant to existing treatment, pharmaceutical development of these compounds has lagged. Thus, the current review focuses on the key algal sources that have been exploited as anticancer therapeutic leads, including their biological origins, phytochemistry, and the challenges involved in converting such leads into effective anticancer drugs.


Assuntos
Antineoplásicos , Produtos Biológicos , Neoplasias , Humanos , Proteômica , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias/tratamento farmacológico , Desenvolvimento de Medicamentos , Plantas , Produtos Biológicos/farmacologia , Produtos Biológicos/uso terapêutico
14.
FEMS Microbiol Lett ; 369(1)2022 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-35675219

RESUMO

Astronauts undergo space travel to bring scientific information to benefit humanity under various missions of space agencies such as NASA, European Space Agency, Indian Space Research Organization etc. During space missions, they encounter several stressors namely microgravity, fluid shifts, cosmic radiation, sleep deprivation and alteration in the circadian rhythm perturbing the quality of sleep. In addition, confined spaces makes pathogen interaction more likely if a pathobiont gets introduced into spacecraft. Microbiota is the first line оf resistаnсe tо vаriоus disorders and diseаses. It direсtly influenсes the biосhemiсаl, рhysiоlоgiсаl, аnd immunоlоgiсаl раthwаys. 'Gut microbiota' is essential for maintenance of healthy gut barrier functions. 'Dysbiosis' refers to perturbation of microbiota which is correlated with several metabolic and psychological disorders. Microbial metabolites are implicated in maintenance of human health. Investigations conducted on astronauts in international space missions and on analog terrestrial models have indicated a 'dysbiosis' of the gut microbiota associated with spaceflights. 'Dysbiosis' of the gut microbiome observed in astronauts has been implicated in immune dysregulation and a probiotic enriched diet is proposed to restore immune homeostasis. This article not just summarizes the state of art research on dysbiosis of the gut microbiome of astronauts, but also a diet mediated correction plan to restore their health especially during long term space missions. A characterization of microbial metabolites of the gut to enable administration of astronaut specific probiotic, postbiotic or synbiotic to alleviate space associated dysbiosis is proposed. It is also recommended that astronauts maintain a balanced nutritious diet throughout life to promote a resilient microbiota that is not perturbed by space missions. Further, a bioregenerative life support system wherein a probiotic may be produced in space station is proposed.


Assuntos
Microbiota , Voo Espacial , Astronautas , Dieta , Disbiose , Humanos , Astronave
15.
J Biotechnol ; 355: 21-41, 2022 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-35752390

RESUMO

Plant nutrition is crucial in crop productivity and providing food security to the ever-expanding population. Application of chemical/biological fertilizers and pesticides are the mainstays for any agricultural economy. However, there are unintended consequences of using chemical fertilizers and pesticides. The environment and ecological balance are adversely affected by their usage. Biofertilizers and biopesticides counter some undesired environmental effects of chemical fertilizers/pesticides; despite some drawbacks associated with their use. The recent developments in nanotechnology offer promise toward sustainable agriculture. Sustainable agriculture involves addressing the concerns about agriculture as well as the environment. This review briefs about important nanomaterials used in agriculture as nanofertilizers, nanopesticides, and a combination called nanobiofertilizers. Both nanofertilizers and nanopesticides enable slow and sustained release besides their eco-friendly nature. They can be tailored to the specific needs of to crop. Nanofertilizers also offer greater stress tolerance and, therefore, are of considerable value in the era of climate change. Furthermore, nanofertilizers/nanopesticides are applied in minute amounts, reducing transportation costs associated and thus positively affecting the economy. Their uses extend beyond such as if nanoparticles (NPs) are used at high concentrations; they affect plant pathogens adversely. Polymer-based biodegradable nanofertilizers and nanopesticides offer various benefits. There is also a dark side to the use of nanomaterials in agriculture. Nanotechnology often involves the use of metal/metal oxide nanoparticles, which might get access to human bodies leading to their accumulation through bio-magnification. Although their effects on human health are not known, NPs may reach toxic concentrations in soil and runoff into rivers, and other water bodies with their removal to become a huge economic burden. Nevertheless, a risk-benefit analysis of nanoformulations must be ensured before their application in sustainable agriculture.


Assuntos
Fertilizantes , Praguicidas , Agricultura , Produção Agrícola , Fertilizantes/análise , Humanos , Nanotecnologia , Plantas
16.
Lab Chip ; 22(8): 1519-1530, 2022 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-35266489

RESUMO

Circulating tumor cells (CTCs) are distinct cancer biomarkers established in clinical settings for early cancer detection, metastasis progression, and minimal residual disease (MRD) monitoring. Despite numerous advances, the comprehensive molecular characterization of CTCs is extremely challenging owing to their rarity and heterogeneity. Here, we present a novel cotton microfluidic substrate (CMS) as an innovative biomedical matrix that efficiently isolates CTCs while facilitating in vitro CTC expansion to enable a further downstream analysis of these rare cells. CMS enabled static and dynamic isolation of cells from the MCF-7 cancer cell line, as well as from head and neck squamous cell carcinoma (HNSCC) patients' blood and the cell capture efficiencies were further compared with a clinically regulated OncoDiscover® Liquid Biopsy Test. Further, CMS acted as a matrix on which the captured cancer cells were grown in 3D tumor models for studying anti-cancer drug efficacy and multi-drug resistance (MDR) mechanisms. The design of the CMS employed two different surface chemistries, flattened and nanostructured surfaces, each conjugated to anti-EpCAM antibodies to evaluate the CTC capture efficiency and 3D tumor growth dynamics. The nanostructured surface was highly efficient for capturing CTCs and promoted 3D tumor spheroid formation with a 5-fold increase in size from day 03 to day 10 of culture. Moreover, when treated with an anti-cancer drug, cisplatin, an almost 1/2 reduction in tumor size was achieved within 24 hours, followed by a cytostatic threshold and eventual acquisition of drug resistance within 3 days. Conclusively, the CMS matrix exhibits potential for further development of "tissue on chip" and "point-of-care" medical devices in cancer diagnostics, and chemo-therapeutic efficacy evaluations in both drug discovery and development.


Assuntos
Antineoplásicos , Células Neoplásicas Circulantes , Anticorpos , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Separação Celular , Humanos , Biópsia Líquida , Células Neoplásicas Circulantes/patologia
17.
Artigo em Inglês | MEDLINE | ID: mdl-35595620

RESUMO

OBJECTIVE: The aim of this study was to investigate the presence of circulating tumor cells (CTCs) and their correlation with prognostic factors and clinical outcomes in treatment-naive patients with oral squamous cell carcinoma. STUDY DESIGN: CTCs were isolated using OncoDiscover technique from presurgically obtained peripheral blood of 152 patients with treatment naïve oral squamous cell carcinoma. Sensitivity analysis was performed by including 40 healthy controls. CTCs cutoff values for clinicopathologic factors were obtained from receiver operating characteristic curves. Multivariate models determined the significance of CTC as independent variables. Kaplan-Meier analysis differentiated in overall survival between CTC values corresponding to the stage. RESULTS: Sensitivity, specificity, and accuracy of CTC detection were 94.32%, 98%, and 95.17%, respectively. Platform differentiated true positives at >3.5 CTCs (P < .00001). CTCs above 20.5 were suggestive of nodal metastasis (P < .0001) with a linear trend for detecting occult metastasis (P = .061). Early and advanced stages could be differentiated by >13.5 CTCs (P < .0001). Elevated CTCs were significantly associated with extranodal extension (>21.45 CTCs, P = .025), perineural invasion (>19.35 CTCs, P = .049), and depth of invasion (>12.5 CTCs, P = .0038). Median survival was reduced by 19 months when CTCs were >13. CONCLUSIONS: Preoperative CTC levels demonstrated a strong correlation with adverse clinicopathology factors and suggested its role as a sensitive prognostic marker to predict survival outcome and disease progress.


Assuntos
Carcinoma de Células Escamosas , Neoplasias de Cabeça e Pescoço , Neoplasias Bucais , Células Neoplásicas Circulantes , Biomarcadores Tumorais , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/terapia , Humanos , Neoplasias Bucais/terapia , Células Neoplásicas Circulantes/patologia , Prognóstico , Carcinoma de Células Escamosas de Cabeça e Pescoço
18.
J Mater Chem B ; 9(13): 2946-2978, 2021 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-33480960

RESUMO

Advanced materials and chemo-specific designs at the nano/micrometer-scale have ensured revolutionary progress in next-generation clinically relevant technologies. For example, isolating a rare population of cells, like circulating tumor cells (CTCs) from the blood amongst billions of other blood cells, is one of the most complex scientific challenges in cancer diagnostics. The chemical tunability for achieving this degree of exceptional specificity for extra-cellular biomarker interactions demands the utility of advanced entities and multistep reactions both in solution and in the insoluble state. Thus, this review delineates the chemo-specific substrates, chemical methods, and structure-activity relationships (SARs) of chemical platforms used for isolation and enumeration of CTCs in advancing the relevance of liquid biopsy in cancer diagnostics and disease management. We highlight the synthesis of cell-specific, tumor biomarker-based, chemo-specific substrates utilizing functionalized linkers through chemistry-based conjugation strategies. The capacity of these nano/micro substrates to enhance the cell interaction specificity and efficiency with the targeted tumor cells is detailed. Furthermore, this review accounts for the importance of CTC capture and other downstream processes involving genotypic and phenotypic CTC analysis in real-time for the detection of the early onset of metastases progression and chemotherapy treatment response, and for monitoring progression free-survival (PFS), disease-free survival (DFS), and eventually overall survival (OS) in cancer patients.


Assuntos
Biomarcadores Tumorais/análise , Neoplasias/diagnóstico , Células Neoplásicas Circulantes/patologia , Humanos
19.
ACS Appl Mater Interfaces ; 12(51): 56839-56849, 2020 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-33290035

RESUMO

Magnetoplasmonic nanomaterials, which combine light and magnetic field responsiveness in an advantageous manner, are attractive candidates for bio-nanoapplications. However, the synthetic access to such hybrid particles has been limited by the incompatibility of the iron- and gold-based lattices. In this work, we provide the first insights into a new synthetic strategy for developing magnetoplasmonic anisotropic nanocomposites with prominent phototransducing properties. In our approach, magnetic nanocubes based on an alloy of iron oxide, zinc, and silver were constructed. In a key second stage, the galvanic replacement of silver with gold atoms yielded satellite-like magnetoplasmonic anisotropic structures. Superior magnetic and photoconverting properties were observed for the novel magnetoplasmonic nanocomposites when compared with the pure parent structures. Moreover, the synergy between the magnetic and optical stimuli was examined, showing shape-dependent contributions in the magnetization experiments. More importantly, an excellent cell ablation capability upon laser irradiation was observed for the magnetoplasmonic nanocomposites compared to the pure magnetic or plasmonic controls. Further demonstration of these novel theragnostic agents as MRI contrast agents is also reported even during the light-irradiation event. Thus, the described particles showed promising properties for bioapplications emerging from the novel synthetic methodology.

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