Aptamer-Modified Nb2C Multifunctional Nanomedicine for Targeted Photothermal/Chemotherapy Combined Therapy of Tumor.

The poor delivery efficiency of nanotherapeutic drugs and their potential off-target toxicity significantly limit their effectiveness and extensive application. An active targeting system with high efficiency and few side effects is a promising strategy for tumor therapy. Herein, a multifunctional nanomedicine Nb2C-PAA-DOX@Apt-M (NDA-M) was constructed for targeted photothermal/chemotherapy (PTT/CHT) combined tumor therapy. The specific targeting ability of aptamer could effectively enhance the absorption of nanomedicine by the MCF-7 cell. By employing Apt-M, the NDA-M nanosheets demonstrated targeted delivery to MCF-7 cells, resulting in enhanced intracellular drug concentration. Under 1060 nm laser irradiation, a rapid temperature increase of the NDA-M was observed within the tumor region to achieve PTT. Meanwhile, CHT was triggered when DOX release was induced by photothermal/acid stimulation. The experimental results demonstrated that aptamer-mediated targeting achieved enhanced PTT/CHT efficacy both in vitro and in vivo. Notably, NDA-M induced complete ablation of solid tumors without any adverse side effects in mice. This study demonstrated new and promising tactics for the development of nanomaterials for targeted tumor therapy.

Electrospun Aspirin/Eudragit/Lipid Hybrid Nanofibers for Colon-targeted Delivery Using an Energy-saving Process.

Both electrospinning apparatus and their commercial products are extending their applications in a wide variety of fields. However, very limited reports can be found about how to implement an energy-saving process and in turn to reduce the production cost. In this paper, a brand-new type of coaxial spinneret with a solid core and its electrospinning methods are developed. A novel sort of medicated Eudragit/lipid hybrid nanofibers are generated for providing a colon-targeted sustained release of aspirin. A series of characterizations demonstrates that the as-prepared hybrid nanofibers have a fine linear morphology with the aspirin/lipid separated from the matrix Eudragit to form many tiny islands. In vitro dissolution tests exhibit that the hybrid nanofibers are able to effectively prevent the release of aspirin under an acid condition (8.7%±3.4% for the first two hours), whereas prolong the drug release time period under a neutral condition(99.7±4.2% at the seventh hour). The energy-saving mechanism is discussed in detail. The prepared aspirin-loaded hybrid nanofibers can be further transferred into an oral dosage form for potential application in countering COVID-19 in the future.

Long noncoding RNA LBX2-AS1-modulated miR-4766-5p regulates gastric cancer development through targeting CXCL5.

BACKGROUND: Long noncoding RNAs (LncRNAs) have been reported to critically regulate gastric cancer (GC). Recently, it was reported that LBX2 antisense RNA 1 (LBX2-AS1) is abnormally expressed in GC. However, the role of LBX2-AS1 in the malignancy of GC is worth further discussion. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the LBX2-AS1, miR-4766-5p and C-X-C motif chemokine (CXCL5) expression in GC tissues and cells. Dual-luciferase reporter assay was applied to examine the target relationship between LBX2-AS1 and miR-4766-5p or miR-4766-5p and CXCL5. Cell counting kit-8 (CCK-8) and Transwell assays were used to detect cell proliferation, migration and invasion rates. The protein expression of CXCL5 was confirmed using western blot. The RNA pull down experiment was used to verify the specificity of LBX2-AS1 and miR-4766-5p on BGC-823 and SGC-7901 cells. RESULTS: LBX2-AS1 was up-regulated in GC tissues and cells, and its knockdown suppressed proliferation, migration and invasion of GC cells. While, overexpression of LBX2-AS1 increased proliferation and increased CXCL5 mRNA level. CXCL5 improved cell proliferation, migration and invasion of GC cells. LBX2-AS1 could bind to miR-4766-5p to regulate CXCL5 expression. Overexpression of CXCL5 overturned those effects of miR-4766-5p in GC cells. RNA Pull down shown that BGC-823 and SGC-7901 cells, miR-4766-5p specifically binds to LBX2-AS1. CONCLUSIONS: In short, this study demonstrated that LBX2-AS1 promoted proliferation, migration and invasion through up-regulation CXCL5 mediated by miR-4766-5p in GC. The LBX2-AS1/miR-4766-5p/CXCL5 regulatory axis provides a theoretical basis for the research on lncRNA-directed therapeutics in GC.

Capillary electrophoretic study of green fluorescent hollow carbon nanoparticles.

CE coupled with laser-induced fluorescence and UV absorption detections has been applied to study the complexity of as-synthesized green fluorescent hollow carbon nanoparticles (HC-NP) samples. The effects of pH, type, and concentration of the run buffer and SDS on the separation of HC-NP are studied in detail. It is observed that phosphate run buffer is more effective in separating the HC-NP and the optimal run buffer is found to be 30 mM phosphate and 10 mM SDS at pH 9.0. The CE separation of this HC-NP is based on the difference in size and electrophoretic mobility of HC-NP. Some selected HC-NP fractions are collected and further characterized by UV-visible absorption and photoluminescence (PL) spectroscopy, MS, and transmission electron microscopy. The fractionated HC-NP show profound differences in absorption, emission characteristics, and PL quantum yield that would have been otherwise misled by studying the complex mixture alone. It is anticipated that our CE methodology will open a new initiative on extensive studies of individual HC-NP species in the biomedical, catalysis, electronic, and optical device, energy storage, material, and sensing field.

A turn-on fluorescent aptasensor for adenosine detection based on split aptamers and graphene oxide.

A simple, sensitive and selective turn-on fluorescent aptasensor for adenosine detection was developed based on target-induced split aptamer fragment conjunction and different interactions of graphene oxide and the two states of the designed aptamer sequences.

Goat anti-rabbit IgG conjugated fluorescent dye-doped silica nanoparticles for human breast carcinoma cell recognition.

We report an indirect method for cancer cell recognition using photostable fluorescent silica nanoprobes as biological labels. The dye-doped fluorescent silica nanoparticles were synthesized using the water-in-oil (W/O) reverse microemulsion method. The silica matrix was produced by the controlled hydrolysis of tetraethylorthosilicate (TEOS) in water nanodroplets with the initiation of ammonia (NH3·H2O). Fluorescein isothiocyanate (FITC) or rhodamine B isothiocyanate conjugated with dextran (RBITC-Dextran) was doped in silica nanoparticles (NPs) with a size of 60 ± 5 nm as a fluorescent signal element by covalent bonding and steric hindrance, respectively. The secondary antibody, goat anti-rabbit IgG, was conjugated on the surface of the PEG-terminated modified FITC-doped or RBITC-Dextran-doped silica nanoparticles (PFSiNPs or PBSiNPs) by covalent binding to the PEG linkers using the cyanogen bromide method. The concentrations of goat anti-rabbit IgG covering the nanoprobes were quantified via the Bradford method. In the proof-of-concept experiment, an epithelial cell adhesion molecule (EpCAM) on the human breast cancer SK-Br-3 cell surface was used as the tumor marker, and the nanoparticle functionalized with rabbit anti-EpCAM antibody was employed as the nanoprobe for cancer cell recognition. Compared with fluorescent dye labeled IgG (FITC-IgG and RBITC-IgG), the designed nanoprobes display dramatically increased stability of fluorescence as well as photostability under continuous irradiation.

Co-assembly of graphene/polyoxometalate films for highly electrocatalytic and sensing hydroperoxide.

Graphene oxide (GO) films mixed with polyethylenimine (PEI) were prepared by a layer-by-layer assembly (LBL) method, in which the GO component is then converted to reduced GO (rGO) in situ through an electron transfer interaction with a polyoxometalate (POM) that is assembled on the outer surface. With this, devices were manufactured by spreading composite films of (PEI/rGO)n-POM with different numbers of PEI/rGO layers on ITO substrates. Cyclic voltammetry (CV) reveals that the catalytic activity for H2O2 of (PEI/rGO)n-POM films was significantly higher than that of similar films of (PEI/GO)n/PEI/POM manufactured LBL with the same number of layers, although the catalyst POM content of (PEI/rGO)n-POM was only half that of (PEI/GO)n/PEI/POM. The catalytic activity of (PEI/rGO)n-POM films first increases and then decreases as the number of PEI/rGO layers increases. The result shows that (PEI/rGO)3-POM films with three PEI/rGO layers exhibit the highest efficiency. Amperometric measurements of the (PEI/rGO)3-POM films showed improved current response, high sensitivity, wide linear range, low detection limit, and fast response for H2O2 detection. The enhanced catalytic property of (PEI/rGO)n-POM films is attributed to the electron transfer interaction and electrostatic interaction between POM and rGO.

A novel ratiometric fluorescent probe based on dual-emission carbon dots for highly sensitive detection of salicylic acid.

In this study, a novel ratiometric fluorescence probe based on dual-emission carbon dots (CDs) for the sensitive detection of salicylic acid (SA) was constructed for the first time. The dual-emission CDs were synthesized by simple hydrothermal method using tartaric acid (TA) and m-phenylenediamine (mPD) as raw materials. In the presence of SA, the fluorescence intensity of CDs was enhanced at 499 nm, but remained basically unchanged at 439 nm. This phenomenon is caused by the intermolecular hydrogen bond interactions. The concentrations of SA had an excellent linear relationship with CDs' fluorescence intensity ratio (F499/F439) in a range of 1 âˆ¼ 120 and 120 âˆ¼ 240 µM with low detection limits of 0.68 and 1.05 µM. The established ratiometric fluorescent probe is economical, simple and green, and can be used for the effective detection of SA. In addition, the proposed ratiometric fluorescent probe was successfully used to monitor SA in facial mask and toning lotion samples with a satisfactory recovery of 99.7-106.7 %. The results show that the constructed fluorescent probe based on dual-emission CDs has a great potential for the rapid and sensitive analysis of SA in actual samples.

The lncRNA-miRNA-integrin alpha V ceRNA network can affect the occurrence and prognosis of gastric cancer.

OBJECTIVES: The aim of this study was to explore the role of integrin alpha V (ITGAV) and the related long noncoding RNA-microRNA-messenger RNA competing endogenous RNA (lncRNA-miRNA-mRNA ceRNA) network in the development and prognosis of cancers, especially gastric cancer (GC), through bioinformatic analysis. METHODS: Pan-cancer and GC data were collected from the UCSC Xena website, and validation datasets were obtained from the Gene Expression Omnibus (GEO). R (version 3.6.3), GraphPad Prism 8, and SPSS 23.0 software were used to analyze data and prepare figures. RESULTS: The expression of ITGAV in tumor tissues was higher than that of normal tissues in ten cancer types. A lower expression of ITGAV in five tumors (CESC, LGG, LIHC, MESO, and STAD) predicted better patient prognosis. In GC, the mRNA and protein expression of ITGAV in tumor tissues was higher than that of normal tissues. Patients with high ITGAV expression had poor prognosis and clinical characteristics, including worse grades and more advanced stages. Patients with higher ITGAV expression had higher immune and stromal scores and lower purity (P<0.05). In addition, seven miRNAs were found that were negatively correlated with ITGAV expression through the website; high expression of these miRNAs indicated a better prognosis. Using this correlation, the authors built the lncRNA-miRNA-ITGAV ceRNA network, to predict the prognosis of GC. CONCLUSIONS: This study showed that ITGAV could be considered a prognostic factor for GC, and an lncRNA-miRNA-ITGAV ceRNA network was built to promote the exploration of the mechanism and prognosis of GC.

Effects of Graphene Oxide on Plant Growth: A Review.

Several reports of graphene oxide (GO) promoting plant growth have sparked interest in its potential applications in agroforestry. However, there are still some toxicity studies that have raised concerns about the biosafety of GO. These reports show conflicting results from different perspectives, such as plant physiology, biochemistry, cytology, and molecular biology, regarding the beneficial and detrimental effects of GO on plant growth. Seemingly inconsistent studies make it difficult to effectively apply GO in agroforestry. Therefore, it is crucial to review and analyze the current literature on the impacts of GO on plant growth and its physiological parameters. Here, the biological effects of GO on plant growth are summarized. It is proposed that an appropriate concentration of GO may be conducive to its positive effects, and the particle size of GO should be considered when GO is applied in agricultural applications. This review provides a comprehensive understanding of the effects of GO on plant growth to facilitate its safe and effective use.

Simulations and error analysis of the CNC milling of a face gear tooth with given tool paths.

This data article gives the validation files to the article "CNC milling of face gears with a novel geometric analysis" [1]. The data is about the simulation and machining error analysis of the CNC milling of a face gear tooth with given tool paths. It includes four files. Three of them are simulation videos of the CNC milling process in VERICUT with a general view, partial view and enlarged view, respectively. The other one is the source file of the machining error analysis, and it has the design model of the face gear, the simulated machined model of the face gear, and machining error analysis according to the comparison of the design model and simulated machined model.

[Investigation of interaction between levofloxacin and bovine serum albumin by fluorescence analysis based on liquid drop].

The interaction between pharmaceutical and protein is an important pharmacokinetic characteristic. Most kinds of drugs must reach the receptor to perform the pharmacological function by plasma. Albumins can serve as a depot protein and a transport protein for numerous endogenous and exogenous compounds. It is of great significance to investigate the binding interaction between albumin and drugs, since it can not only help understand the transportation and distribution of drugs but also elucidate the mechanism. Under the physiological condition of body, the interaction between levofloxacin (LVFX) and bovine serum albumin (BAS) was investigated by fluorescence spectra and ultraviolet absorbance (UV) spectra based on liquid drop. The experimental results showed that LVFX quenches the fluorescence of BAS by forming a LVFX-BSA complex. According to Lineweaver-Burk equation, the apparent binding constants (K(LB)) between LVFX and BSA were 1.694 9 x 10(5) L x mol(-1) (291 K) and 2.881 0 x 10(5) L x mol(-1) (310 K), and the binding sites (n) were 0.884 9 (291 K) and 0.808 9 (310 K). Thermodynamic parameters could be evaluated from the thermodynamic second law, with deltaH (enthalpy) being 20.94 kJ mol(-1) and deltaS (entropy) being 172.1 J x mol(-1). According to the relation between thermodynamic parameters and the interaction force, LVFX depended principally on the hydrophobic interaction to bind with BSA. The results showed that the quenching belonged to static fluorescence quenching with non-radiation energy transfer happening within single molecule. The binding locality was an area 2.68 nm away from tryptonphan residue-212 in BAS according to Forster's non-radiation energy transfer mechanism. The conformational changes of BSA were evaluated by measuring the synchronous fluorescence intensity of protein amino acid residues, both before and after the addition of LVFX. A slightly stronger blue-shift of tryptophan fluorescence upon the addition of drug was observed, and the emission maximum of tyrosine kept its position. It was suggested that the environments of tryptophan residues in pure albumin solution are relatively polar. Binding of LVFX changes the environments to apolar ones. The shift in polarity is brought about by confirmation changes due to the interaction between albumin and ligand molecule.

A novel highly fluorescent S, N, O co-doped carbon dots for biosensing and bioimaging of copper ions in live cells.

In this study, novel highly fluorescent sulfur, nitrogen, and oxygen co-doped carbon dots (S, N, O-CDs) were prepared from m-phenylenediamine and sulfamide by using the hydrothermal method. The prepared S, N, O-CDs show high doping rate and fluorescence yield as well as long-term fluorescence stability. In addition, S, N, O-CDs show good fluorescence response towards Cu2+ over a concentration range of 2-60 µM with a detection limit of 0.29 µM. Taking advantages of the properties of S, N, O-CDs including high selectivity and low cytotoxicity, S, N, O-CDs were successfully applied for Cu2+ sensing and imaging in the cells and O2˙--induced Cu2+ increase in the cells was observed. Furthermore, on account of strong complexation between Cu2+ and pyrophosphate ion (PPi) as well as specific hydrolysis ability of alkaline phosphatase (ALP) towards PPi, PPi and ALP were further detected with high selectivity based on S, N, O-CDs/Cu2+ system. The prepared S, N, O-CDs showed good detection results for PPi and ALP with detection limits of 0.44 µM and 1.03 U L-1, respectively. Moreover, the developed method also realized PPi and ALP detection in real samples.

Indirect immunofluorescence detection of E. coli O157:H7 with fluorescent silica nanoparticles.

A method of fluorescent nanoparticle-based indirect immunofluorescence assay using either fluorescence microscopy or flow cytometry for the rapid detection of pathogenic Escherichia coli O157:H7 was developed. The dye-doped silica nanoparticles (NPs) were synthesized using W/O microemulsion methods with the combination of 3-aminopropyltriethoxysilane (APTES) and fluorescein isothiocyanate (FITC) and polymerization reaction with carboxyethylsilanetriol sodium salt (CEOS). Protein A was immobilized at the surface of the NPs by covalent binding to the carboxyl linkers and the surface coverage of Protein A on NPs was determined by the Bradford method. Rabbit anti-E. Coli O157:H7 antibody was used as primary antibody to recognize E. coli O157:H7 and then antibody binding protein (Protein A) labeled with FITC-doped silica NPs (FSiNPs) was used to generate fluorescent signal. With this method, E. Coli O157:H7 in buffer and bacterial mixture was detected. In addition, E. coli O157:H7 in several spiked background beef samples were measured with satisfactory results. Therefore, the FSiNPs are applicable in signal-amplified bioassay of pathogens due to their excellent capabilities such as brighter fluorescence and higher photostability than the direct use of conventional fluorescent dyes.

Sensitive determination of kaempferol using carbon dots as a fluorescence probe.

In this study, a new sensitive and convenient method for the determination of kaempferol (Kae) based on the fluorescence quenching of fluorescent carbon dots (C-dots) was developed. The C-dots were prepared by simply mixing acetic acid, water and diphosphorus pentoxide. This green synthesis approach proceeds rapidly and gives large quantities of C-dots. The fluorescence of the C-dots decreased obviously with the increase of Kae concentration. The effect of other interfering substances on the fluorescence intensity of C-dots showed low interference. Under the optimum conditions, a linear correlation was established between fluorescence intensity ratio Fo/F and the concentration of Kae in the range of 3.5-49 µM with a detection limit (S/N=3) of 38.4 nM. The proposed method has been successfully applied to determination of Kae in xindakang tablets and human serum samples with recovery in the range of 94.6-109.0%. The C-dots could be a promising fluorescence probe for the detection of Kae owing to its low-cost production, easy operation, low cytotoxicity, and excellent biocompatibility.

Sensitive detection of carcinoembryonic antigen using surface plasmon resonance biosensor with gold nanoparticles signal amplification.

A new method for real-time detection of carcinoembryonic antigen (CEA) in human serum with high sensitivity and selectivity using surface plasmon resonance (SPR) biosensor was developed. Two kinds of antibodies were used to recognize CEA at different epitopes with high affinity and specificity. Gold nanoparticles (GNPs) modified with streptavidin (SA) were used to further enhance signal specifically via biotin-streptavidin interaction. The binding capacity of the streptavidin-modified gold nanoparticles (SA-GNPs) for ligand biotin was quantified by titration with biotin (5-fluorescein) conjugate to be 10.54 biotin binding sites per 100 nm(2). The developed GNPs enhanced sandwich SPR biosensor successfully fulfilled the sensitive detection of CEA in the range of 1-60 ng/mL with a detection limit of 1.0 ng/mL. Compared to the direct assay format, sandwich format without GNPs and SA-GNPs enhanced sandwich format led to 4.2-fold and 13.8-fold in the sensitivity, respectively. This sensor also showed good selectivity for CEA in the interference study. The results demonstrated that the proposed method could provide a high sensitivity and selectivity in the detection of CEA and offer a promising alternative for cancer biomarker than traditional clinical examinations.

MUC-1 aptamer-conjugated dye-doped silica nanoparticles for MCF-7 cells detection.

In this work, we have prepared three types of aptamer-conjugated Rubpy-doped silica nanoparticles for Human breast carcinoma MCF-7 cells labeling. Probe A is prepared through covalent conjugation between amine-labeled MUC-1 aptamer and carboxyl-modified Rubpy-doped NPs (NPs-aptamer). Probe B is prepared based on the interaction between biotin-labeled MUC-1 aptamer and avidin-conjugated Rubpy-doped NPs (NPs-avidin-biotin-aptamer). For Probe C, there is a PEG with flexible long chain as the bridge between avidin and the NPs (NPs-PEG-avidin-biotin-aptamer). In addition, we further investigate the practical number of MUC-1 aptamers on an NP of each probe using hoechst33258 dye. The binding efficiency of MUC-1 aptamer on the three types of probes as follows: Probe A < Probe B < Probe C. In addition, microscopic fluorescence imaging shows that Probe C containing the PEG molecules can be effectively applied for the recognition of MUC-1 protein in human breast carcinoma MCF-7 cells thus demonstrates that the PEG with flexible long chain as the bridge between the aptamer and NP can greatly enhances the freedom of MUC-1 aptamer. Compared with common organic dyes, the dye-doped silica nanoparticles serve as a stable bioprobe because of their facile conjugation with the desirable biomolecules, and have exhibited great potential in bioanalysis.

Covalent conjugation of avidin with dye-doped silica nanopaticles and preparation of high density avidin nanoparticles as photostable bioprobes.

Progress in biomedical imaging depends on the development of bioprobes with a high sensitivity and stability. Fluorescent silica nanoparticles (NPs) covalent conjugation of avidin has been proposed for cancer cells imaging by fluorescence microscopy. Uniform silica NPs were prepared using water-in-oil (W/O) microemulsion methods and primary amine groups were introduced onto the surface of the NPs by condensation of tetraethyl orthosilicate (TEOS). Optically stable organic dyes, tris(2,2'-bipyridyl) dichlororuthenium(II) hexahydrate (Rubpy), were doped inside the silica NPs. The amine functions were transferred to carboxyl groups coupled with a linker elongation. Avidin was immobilized at the surface of the NPs by covalent binding to the carboxyl linkers. The binding capacity of the avidin-covered NPs for ligand biotin was quantified by titration with biotin(5-fluorescein) conjugate to 1.25 biotin binding sites/100 nm(2). We used biotinylated antibody and cell recognition by fluorescence microscopy imaging technique. The lung carcinoma cells were identified easily with high efficiency using these antibody-coated NPs. By comparison with fluorescein isothiocyanate (FITC), dye-doped silica NPs display dramatically increased stability of fluorescence as well as photostability, as compared to the common organic dye, when under continuous irradiation.

Silica nanoparticles based label-free aptamer hybridization for ATP detection using hoechst33258 as the signal reporter.

In this work, we have developed a simple and sensitive method for ATP detection using silica nanoparticles (NPs) as the platform and hoechst33258 as the signal reporter. The ATP-binding aptamers hybridize with the probe DNA (DNA(p)) immobilized NPs to form the aptamer/DNA(p) duplex on the NPs surface. The conformational change of the aptamer leads to the decrease of the aptamer/DNA(p) duplex on the NPs due to the ATP-binding aptamer switches its structure from the aptamer/DNA(p) duplex to the aptamer/target complex in the presence of ATP. ATP detection can be easily realized by separating the silica nanoparticles and adding the hoechst33258 of intercalating to aptamer/DNA(p) (dsDNA). Good selectivity between ATP and CTP, GTP or UTP has been demonstrated, which is due to the specific recognition between ATP aptamer and ATP. The K(d) was estimated to be ∼1mM from 0 to 4mM and a liner response was observed from 0 to 0.2mM with a detection limit of ∼20µM. Compared with other methods, the carboxyl-modified silica nanoparticles (∼60nm) prepared by the reverse microemulsion method can serve as a stable and sensitive sensor platform because of their smaller size and facile conjugation with amine-containing molecules. In addition, the high sensitivity and selectivity of hoechst33258 was employed for the ssDNA and dsDNA determination, which takes advantage of the label-free aptamer and lower cost.

Evaluation of the bioconjugation efficiency of different quantum dots as probes for immunostaining tumor-marker proteins.

The differing bioconjugation efficiencies of quantum dots (QDs) are a practical obstacle to their popularization. Differences in bioconjugation efficiency based on immunostaining the same targeted molecules using different batches of QDs need to be evaluated prior to their application. In this paper, a quantitative method for evaluating the efficiency of QDs in staining tissues has been developed based on Hadamard transform (HT) spectral imaging. Proliferating cell nuclear antigens (PCNA) in breast cancer tissues were labeled with bioconjugated QD bioprobes using a 454 nm laser as the light source for fluorescence spectral imaging. Four-dimensional (4D) spectral imaging analysis of PCNA in cell nuclei was carried out using HT spectral microscopy based on immunostaining with different batches of QDs. The fluorescence intensity distributions in the cell nuclei were collected from the 4D images. Based on the information obtained from microscopic spectra and 4D images, differences in the bioconjugation efficiency among different batches of QDs were evaluated. The results demonstrate that it is possible to maintain uniform bioconjugation efficiencies with different QD bioconjugation processes in order to obtain accurate and reliable results in biomedical analysis and cancer diagnosis.