One of the current issues with thyroid tumor is early diagnosis as it makes the higher possibility of curing. This research was focused to detect and quantify the level of specific target sequence complementation of miR-222 with capture DNA sequence on interdigitated electrode (IDE) sensor. The aluminum electrode with the gap and finger sizes of 10 µm was fabricated on silicon wafer, further the surface was amine-functionalized for accommodating carboxylated-DNA probe. With DNA-target RNA complementation, the detection limit was attained to be 1 fM as estimated by a linear regression analysis [y = 1.5325x - 2.1171 R² = 0.9065] and the sensitivity was at the similar level. Current responses were higher by increasing the target RNA sequence concentrations. Control experiments with mismatched/noncomplementary sequences were failed to complement the capture DNA sequence immobilized on IDE, indicating the specific target validation. This research helps diagnosing and identifying the progression with thyroid tumor and miRNA being a potential "marker" in atypia diagnosis.
A high-performance interdigitated electrode (IDE) biosensing surface was reported here by utilizing self-assembled silica nanoparticle (SiNP). The modified surface was used to evaluate the complementation of hairpin forming region from Mitoxantrone resistance gene 7 (MXR7; liver cancer-related short gene). The conjugated SiNPs on 3-aminopropyl triethoxysilane functionalization were captured with probe sequence on IDE biosensing surface. The physical and chemically modified surface was used to quantify MXR7 and an increment in the current response upon complementation was noticed. Limit of target DNA detection was calculated (1-10 fM) and this label-free detection is at the comparable level to the fluorescent-based sensing. A linear regression was calculated [y = 0.243x - 0.0773; R² = 0.9336] and the sensitivity was 1 fM on the linear range of 1 fM to 10 pM. With the strong attachment of capture DNA on IDE through SiNP, the surface clearly discriminates the specificity (complementary) versus nonspecificity (complete-, single-, and triple-mismatched sequences). This detection strategy helps to determine liver cancer progression and the similar strategy can be followed for other gene sequence complementation.
Current developments in sensors and actuators are heralding a new era to facilitate things to happen effortlessly and efficiently with proper communication. On the other hand, Internet of Things (IoT) has been boomed up with er potential and occupies a wide range of disciplines. This study has choreographed to design of an algorithm and a smart data-processing scheme to implement the obtained data from the sensing system to transmit to the receivers. Technically, it is called "telediagnosis" and "remote digital monitoring," a revolution in the field of medicine and artificial intelligence. For the proof of concept, an algorithmic approach has been implemented for telediagnosis with one of the degenerative diseases, that is, Parkinson's disease. Using the data acquired from an improved interdigitated electrode, sensing surface was evaluated with the attained sensitivity of 100 fM (n = 3), and the limit of detection was calculated with the linear regression value coefficient. By the designed algorithm and data processing with the assistance of IoT, further validation was performed and attested the coordination. This proven concept can be ideally used with all sensing strategies for immediate telemedicine by end-to-end communications.
Prostate cancer is one of the predominant cancers affecting men and has been widely reported. In the past, various therapies and drugs have been proposed to treat prostate cancer. Among these treatments, gene therapy has been considered to be an optimal and widely applicable treatment. Furthermore, due to the increased specificity of gene sequence complementation, the targeted delivery of complementary gene sequences may represent a useful treatment in certain instances. Various gene therapies, including tumor-suppressor gene therapy, suicide gene therapy, immunomodulation gene therapy and anti-oncogene therapies, have been established to treat a wide range of diseases, such as cardiac disease, cystic fibrosis, HIV/AIDS, diabetes, hemophilia, and cancers. To this end, several gene therapy clinical trials at various phases are underway. This overview describes the developments and progress in gene therapy, with a special focus being placed on prostate cancer.
Artificial intelligence of things (AIoT) has become a potential tool for use in a wide range of fields, and its use is expanding in interdisciplinary sciences. On the other hand, in a clinical scenario, human blood-clotting disease (Royal disease) detection has been considered an urgent issue that has to be solved. This study uses AIoT with deep long short-term memory networks for biosensing application and analyzes the potent clinical target, human blood clotting factor IX, by its aptamer/antibody as the probe on the microscaled fingers and gaps of the interdigitated electrode. The earlier results by the current-volt measurements have shown the changes in the surface modification. The limit of detection (LOD) was noticed as 1 pM with the antibody as the probe, whereas the aptamer behaved better with the LOD at 100 fM. The time-series predictions from the AIoT application supported the obtained results with the laboratory analyses using both probes. This application clearly supports the results obtained from the interdigitated electrode sensor as aptamer to be the better option for analyzing the blood clotting defects. The current study supports a great implementation of AIoT in sensing application and can be followed for other clinical biomarkers.
Despite a lot of intensive research on cells-scaffolds interaction, focused are mainly on the capacity of construct scaffolds to regulate cell mobility, migration and cytotoxicity. The effect of the scaffold's topographical and material properties on the expression of biologically active compounds from stem cells is not well understood. In this study, the influence of cellulose acetate (CA) on the electrospinnability of gelatin and the roles of gelatin-cellulose acetate (Ge-CA) on modulating the release of biologically active compounds from amniotic fluid stem cells (AFSCs) is emphasized. It was found that the presence of a small amount of CA could provide a better microenvironment that mimics AFSCs' niche. However, a large amount of CA exhibited no significant effect on AFSCs migration and infiltration. Further study on the effect of surface topography and mechanical properties on AFSCs showed that the tailored microenvironment provided by the Ge-CA scaffolds had transduced physical cues to biomolecules released into the culture media. It was found that the AFSCs seeded on electrospun scaffolds with less CA proportions has profound effects on the secretion of metabolic compounds compared to those with higher CA contained and gelatin coating. The enhanced secretion of biologically active molecules by the AFSCs on the electrospun scaffolds was proven by the accelerated wound closure on the injured human dermal fibroblast (HDF) model. The rapid HDF cell migration could be anticipated due to a higher level of paracrine factors in AFSCs media. Our study demonstrates that the fibrous topography and mechanical properties of the scaffold is a key material property that modulates the high expression of biologically active compounds from the AFSCs. The discovery elucidates a new aspect of material functions and scaffolds material-AFSCs interaction for regulating biomolecules release to promote tissue regeneration/repair. To the best of our knowledge, this is the first report describing the scaffolds material-AFSCs interaction and the efficacy of scratch assays on quantifying the cell migration in response to the AFSCs metabolic products. This article is protected by copyright. All rights reserved.
Plain and Lycium barbarum yogurt were made in the presence and absence of fish collagen. Yogurt samples were analyzed for acidification, milk protein proteolysis, angiotensin I-converting enzyme (ACE) inhibitory activity, and sensory evaluation during refrigerated storage for up to 21 days. The o-phthaldialdehyde peptides amount of L. barbarum yogurt both in the presence and absence of fish collagen were significantly increased during 14 days of storage. SDS-PAGE showed improvement in whey proteins degradation of L. barbarum yogurt with/without fish collagen after 3 weeks of storage. L. barbarum yogurt in absence of fish collagen was acting as a great ACE inhibitor reached up to 85% on day 7 of storage. The incorporation of L. barbarum and/or fish collagen affected to a small extent the overall sensory characteristics of yogurt. Yogurt supplemented with L. barbarum and/or fish collagen may lead to the improvement in the production and formulation of yogurt differing in their anti-ACE activity.
C-reactive protein (CRP) is an acute phase reactant to be a marker of inflammation and has been correlated with the cardiac injury. An immunoassay was performed using anti-human CRP antibody on an InterDigitated electrode (IDE) sensor to determine and specify CRP concentration for diagnosing the condition of myocardial inflammation. To promote the detection, gold nanoparticle (GNP) was seeded on the aminated-IDE surface. Anti-CRP was hitched on the GNP-seeded surface and identified the abundance of CRP. The limit of quantification was found as 100 fM, and the higher current response was noticed by increasing CRP concentrations with the sensitivity at 1 pM. Furthermore, CRP-spiked human serum did not interfere the determination of CRP and increased the current response, indicating suitability for a real-life sample. Similarly, the control experiments with nonimmune antibody Troponin I are not showing the definite current responses, proving the selective identification of CRP. This method of diagnosing is needful to determine the cardiovascular injury at the right time.
A microbial fuel cell is a sustainable and environmental-friendly device that combines electricity generation and wastewater treatment through metabolic activities of microorganisms. However, low power output from inadequate electron transfer to the anode electrode hampers its practical implementation. Nanocomposites of oxidized carbon nanotubes and medium-chain-length polyhydroxyalkanoates (mcl-PHA) grafted with methyl acrylate monomers enhance the electrochemical function of electrodes in microbial fuel cell. Extensive polymerization of methyl acrylate monomers within mcl-PHA matrix, and homogenous dispersion of carbon nanotubes within the graft matrix are responsible for the enhancement. Modified electrodes exhibit high conductivities, better redox peak and reduction of cell internal resistance up to 76%. A stable voltage output at almost 700 mV running for 225 H generates maximum power and current density of 351 mW/m2 and 765 mA/m2 , respectively. Superior biofilm growth on modified surface is responsible for improved electron transfer to the anode hence stable and elevated power output generation.
A highly sensitive silica-alumina (Si-Al)-modified capacitive non-Faradaic glucose biosensor was introduced to monitor gestational diabetes. Glucose oxidase (GOx) was attached to the Si-Al electrode surface as the probe through amine-modification followed by glutaraldehyde premixed GOx as aldehyde-amine chemistry. This Si-Al (∼50 nm) modified electrode surface has increased the current flow upon binding of GOx with glucose. Capacitance values were increased by increasing the glucose concentrations. A mean capacitance value was plotted and the detection limit was found as 0.03 mg/mL with the regression coefficient value, R² = 0.9782 [y = 0.8391x + 1.338] on the linear range between 0.03 and 1 mg/mL. Further, a biofouling experiment with fructose and galactose did not increase the capacitance, indicating the specific glucose detection. This Si-Al-modified capacitance sensor detects a lower level of glucose presence and helps in monitoring gestational diabetes.
This study demonstrated the terminated sialo-sugar chains (Neu5Acα2,6Gal and Neu5Acα2,3Gal) mediated specificity enhancement of influenza virus and chicken red blood cell (RBC) by hemagglutination assay. These glycan chains were immobilized on the gold nanoparticle (GNP) to withhold the higher numbers. With the preliminary optimization, a clear button formation with 0.5% RBC was visualized. On the other hand, intact B/Tokio/53/99 with 750 nM hemagglutinin (HA) displayed a nice hemagglutination. The interference on the specificity of RBC and influenza virus was observed by anti-influenza aptamer at the concentration 31 nM, however, there is no hemagglutination prevention was noticed in the presence of complementary aptamer sequences. Spiking GNP conjugated Neu5Acα2,6Gal or Neu5Acα2,3Gal or a mixture of these two to the reaction promoted the hemagglutination to 63 folds higher with 12 nM virus, whereas under the same condition the heat inactivated viruses were lost the hemagglutination. Neuraminidases from Clostridium perfringens and Arthrobacter ureafaciens at 0.0025 neuraminidase units are able to abolish the hemagglutination. Other enzymes, Glycopeptidase F (Elizabethkingia meningoseptica) and Endoglycosidase H (Streptomyces plicatus) did not show the changes with agglutination. Obviously, sialyl-Gal-terminated glycan conjugated GNP amendment has enhanced the specificity of erythrocyte-influenza virus and able to be controlled by aptamer or neuraminidases. This article is protected by copyright. All rights reserved.
Overuse of antibiotics has led to the development of multi drug resistant strains. Antibiotic resistance is a major drawback in the biomedical field since medical implants are prone to infection by biofilms of antibiotic resistant strains of bacteria. With increasing prevalence of antibiotic resistant pathogenic bacteria, the search for alternative method is utmost importance. In this regard, magnetic nanoparticles are commonly used as a substitute for antibiotics that can circumvent the problem of biofilms growth on the surface of biomedical implants. Iron oxide nanoparticles (IONPs) have unique magnetic properties that can be exploited in various ways in the biomedical applications. IONPs are engineered employing different methods to induce surface functionalization that include the use of polyethyleneimine and oleic acid. IONPs have a mechanical effect on biofilms when in presence of an external magnet. In this review, a detailed description of surface engineered magnetic nanoparticles as ideal antibacterial agents is provided, accompanied by various methods of literature review. This article is protected by copyright. All rights reserved.
Non-small cell lung cancer (NSCLC) incited by epidermal growth factor receptor (EGFR) mutation makes up ∼85% of lung cancer diagnosed and death cases worldwide. The presented study introduced an alternative approach in detecting EGFR mutation using nano-silica integrated with polydimethylsiloxane (PDMS) polymer on interdigitated electrode (IDE) sensor. A 400 μm gap-sized aluminum IDE was modified with nano-polymer layer, which was made up of silica nanoparticles and PDMS polymer. IDE and PDMS-coated IDE (PDMS/IDE) were imaged using electron microscopes that reveals its smooth and ideal sensor morphology. The nano-silica-integrated PDMS/IDE surface was immobilized with EGFR probe and target to specify the lung cancer detection. The sensor specificity was justified through the insignificant current readouts with one-base mismatch and noncomplementary targets. The sensitivity of nano-silica-integrated PDMS/IDE was examined with mutant target spiked in human serum, where the resulting current affirms the detection of EGFR mutation. Based on the slope of the calibration curve, the sensitivity of nano-silica-integrated PDMS/IDE was 2.24E-9 A M-1 . The sensor recognizes EGFR mutation lowest at 1 aM complementary mutant target; however, the detection limit obtained based on 3σ calculation is 10 aM with regression value of 0.97.
Abdominal aortic aneurysm (AAA) is a serious, life-threatening vascular disease that presents as an enlarged area of the aorta, which is the main artery that carries blood away from the heart. AAA may occur at any location in the aorta, but it is mainly found in the abdominal region. A ruptured AAA causes serious health issues, including death. Traditional imaging techniques, such as computed tomography angiogram, magnetic resonance imaging, and ultrasound sonography, have been used to identify AAAs. Circulating biomarkers have recently become attractive for diagnosing AAAs due to their cost-effectiveness compared to imaging. Insulin-like growth factor 1 (IGF-1), a secreted hormone vital for human atherosclerotic plaque stability, has been found to be an efficient biomarker for AAA identification. In this report, immunosensing was performed by using an InterDigitated electrode (IDE) sensor to detect circulating levels of IGF-1. The detection limit of IGF-1 was found to be 100 fM with this sensor. Moreover, related protein controls (IGF-2 and IGFBP3) were not detected with the same antibody, indicating selective IGF-1 detection. Thus, immunosensing by using an IDE sensor may help to effectively diagnose AAAs and represents a basic platform for further development.
One-pot synthesis of sugar-functionalized oligomeric caprolactone was carried out by lipase-catalyzed esterification of ε-caprolactone (ECL) with methyl-d-glucopyranoside (MGP) followed by the elongation of functionalized oligomer chain. Functionalization was performed in a custom-fabricated glass reactor equipped with Rushton turbine impeller and controlled temperature at 60 °C using tert-butanol as reaction medium. The overall reaction steps include MGP esterification of ECL monomer and its subsequent elongation by free 6-hydroxyhexanoate monomer units. A ping-pong bi-bi mechanism without ternary complex was proposed for esterification of ECL and MGP with apparent values of kinetic constant, namely maximal velocity (Vmax ), Michaelis constant for MGP (KmMGP ), and Michaelis constant for ECL (KmECL ) at 3.848 × 10-3 M H-1 , 8.189 × 10-2 M, and 6.050 M, respectively. Chain propagation step of MGP-functionalized ECL oligomer exhibits the properties of living polymerization mechanism. Linear relationship between conversion (%) and number average molecular weight, Mn (g mol-1 ), of functionalized oligomer was observed. Synthesized functionalized oligomer showed narrow range of molecular weight from 1,400 to 1,600 g mol-1 with more than 90% conversion achieved. Structural analysis confirmed the presence of covalent bond between the hydroxyl group in MGP with carboxyl end group of ECL oligomer.
Ovarian cancer starts in the ovaries in its earlier stages and then spreads to the pelvis, uterus, and abdominal region. The success of an ovarian cancer treatment depends on the stage of the cancer and the diagnostic system. Squamous cell carcinoma antigen (SCC-Ag) is one of the most efficient cancer biomarkers, and elevated levels of SCC-Ag in ovarian cancer cells have been used to identify ovarian cancer. Carbon is a potential material for biosensing applications due to its thermal, electrical, and physical properties. Multiwalled carbon nanotubes (MWCNTs) are carbon-based materials that can be used here to detect SCC-Ag. Anti-SCC-Ag antibody was immobilized on the amine-modified MWCNT dielectric sensing surface to detect SCC-Ag. The uniformity of the surface structure was measured with a 3D nanoprofiler, and the results confirmed the detection of SCC-Ag at ∼80 pM. The specific detection of SCC-Ag was confirmed with two control proteins (factor IX and human serum albumin), and the system did not show biofouling. This experimental set-up with MWCNTs a dielectric sensing surface can lead to the detection of ovarian cancer in its initial stages.
Homocysteine [HSCH2 CH2 CH(NH2 )COOH] (Hcy) is a sulfur-containing amino acid of 135.18 Da of molecular weight, generated during conversion of methionine to cysteine. If there is a higher accumulation of Hcy in the blood, that is usually above 15 µmol/L, it leads to a condition referred to as hyperhomocysteinemia. A meta-analysis of observational study suggested an elevated concentration of Hcy in blood, which is termed as the risk factors leading to ischemic heart disease and stroke. Further experimental studies stated that Hcy can lead to an increase in the proliferation of vascular smooth muscle cells and functional impairment of endothelial cells. The analyses confirmed some of the predictors for Hcy presence, such as serum uric acid (UA), systolic blood pressure, and hematocrit. However, angiotensin-converting enzyme inhibitors angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) alone are inadequate for controlling UA and creatinine level, although the addition of folic acid may be beneficial in hypertensive patients who are known to have a high prevalence of elevated Hcy. We hypothesized that combination therapy with an ARB (olmesartan) and folic acid is a promising treatment for lowering the UA and creatinine level in hyperhomocysteinemia-associated hypertension.
Phytosynthesis of gold nanoparticles (AuNPs) has achieved an indispensable significance due to the diverse roles played by biomolecules in directing the physiochemical characteristics of biosynthesized nanoparticles. Therefore, the precise identification of key bioactive compounds involved in producing AuNPs is vital to control their tunable characteristics for potential applications. Herein, qualitative and quantitative determination of key biocompounds contributing to the formation of AuNPs using aqueous Elaeis guineensis leaves extract is reported. Moreover, roles of phenolic compounds and flavonoids in reduction of Au3+ and stabilization of AuNPs have been elucidated by establishing a reaction mechanism. Fourier-transform infrared spectroscopy (FTIR) showed shifting of O─H stretching vibrations toward longer wavenumbers and C═O toward shorter wavenumbers due to involvement of polyphenolic compounds in biosynthesis and oxidation of polyphenolic into carboxylic compounds, respectively, which cape nanoparticles to inhibit the aggregation. Congruently, pyrolysis-gas chromatography-mass spectrometry revealed the major contribution of polyphenolic compounds in the synthesis of AuNPs, which was further endorsed by reduction of total phenolic and total flavonoids contents from 48.08 ± 1.98 to 9.59 ± 0.92 mg GAE/g and 32.02 ± 1.31 to 13.8 ± 0.97 mg CE/g within 60 Min, respectively. Based on experimental results, reaction mechanism explained the roles of phenolic compounds and flavonoids in producing spherical-shaped AuNPs.
Biosynthesis and in vivo depolymerization of intracellular medium-chain-length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 grown on lauric acid were studied. Highest mcl-PHA fraction (>50 % of total biomass) and cell concentration (8 g L-1 ) were obtained at carbon-to-nitrogen (C/N) ratio 20, starting cell concentration 1 g L-1 , and 48 H fermentation. The mcl-PHA comprised of 3-hydroxyhexanoate (C6 ), 3-hydroxyoctanote (C8 ), 3-hydroxydecanoate (C10 ), and 3-hydroxydodecanoate (C12 ) monomers. In vivo action was studied in a mineral liquid medium without carbon source, and in different buffer solutions with varied pH, molarity, ionic strength, and temperature. The monomer liberation rate reflected the mol percentage distribution of the initial polymer subunit composition. Rate and percentage of in vivo depolymerization were highest in 0.2 M Tris-HCl buffer (pH 9, strength = 0.2 M, 30 °C) at 0.21 g L-1 H-1 and 98.6 ± 1.3 wt%, respectively. There is a congruity vis-à-vis to specific buffer type, molarity, pH, ionic strength, and temperature values for superior in vivo depolymerization activities. Direct products from in vivo depolymerization matched the individual monomeric composition of native mcl-PHA. It points to exo-type reaction for the in vivo process, and potential biological route to chiral molecules.
Differentially expressed aqueous soluble proteins between Mycobacterium tuberculosis H37Ra and H37Rv were identified. The protein extracts were separated by two-dimensional gel electrophoresis followed by tandem mass spectrometric analysis. Twelve proteins were detected to be differentially expressed significantly between virulent strain H37Rv and attenuated strain H37Ra. The differentially expression of these proteins was validated by a recently isolated clinical virulent strains of M. tuberculosis, TB138. Out of the 12 proteins identified, which consisted of ten upregulated and two downregulated proteins, nine were belonged to intermediate metabolism and respiration protein group, two were in lipid metabolism, and one protein was involved in information pathways and virulence. Among these proteins, two of the upregulated proteins, namely, mmsA and pntAa, showed a consistent expression pattern in both virulent mycobacterium strains. These proteins can serve as potential biomarkers for the intervention treatment of TB.