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1.
Micromachines (Basel) ; 15(9)2024 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-39337772

RESUMO

Access to clean water is fundamental to public health and safety, serving as the cornerstone of well-being in communities. Despite the significant investments of millions of dollars in water testing and treatment processes, the United States continues to grapple with over 7 million waterborne-related cases annually. This persistent challenge underscores the pressing need for the development of a new, efficient, rapid, low-cost, and reliable method for ensuring water quality. The urgency of this endeavor cannot be overstated, as it holds the potential to safeguard countless lives and mitigate the pervasive risks associated with contaminated water sources. In this study, we introduce a biochip LAMP assay tailored for water source monitoring. Our method swiftly detects even extremely low concentrations of Escherichia coli (E. coli) in water, and 10 copies/µL of E. coli aqueous solution could yield positive results within 15 min on a PC-MEDA biochip. This innovation marks a significant departure from the current reliance on lab-dependent methods, which typically necessitate several days for bacterial culture and colony counting. Our multifunctional biochip system not only enables the real-time LAMP testing of crude E. coli samples but also holds promise for future modifications to facilitate on-site usage, thereby revolutionizing water quality assessment and ensuring rapid responses to potential contamination events.

2.
J Anal Toxicol ; 2024 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-39159049

RESUMO

Occasionally, obtaining an adequate or acceptable postmortem blood specimen for drug analysis is not possible due to factors such as decomposition, exsanguination, or embalming. Submandibular salivary gland tissue, one of three major types of salivary gland tissue in the oral cavity of humans, has been reported to be a viable alternative postmortem specimen for toxicological testing. In this study, we evaluated the performance of the Randox Evidence Investigator instrument and Randox DOA (Drugs of Abuse) Ultra Whole Blood Array for the semi-quantitative determination of 21 immunoassays in an alternative matrix, submandibular salivary gland tissue. We analyzed 132 submandibular salivary gland tissue specimens and compared the generated results to concomitantly collected postmortem whole blood specimen results. Oxycodone 2, meprobamate, barbiturate, benzodiazepine assay 1, zolpidem, and buprenorphine all showed perfect agreement (Cohen's Kappa Score = 1.00) between the submandibular salivary gland tissue results and the postmortem whole blood results; dextromethorphan, fentanyl, benzoylecgonine, methamphetamine, tricyclic antidepressants, oxycodone 1, and opiate showed an almost perfect agreement (Cohen's Kappa Score = 0.81-0.99); methadone, generic opioids, and amphetamine exhibited substantial agreement (Cohen's Kappa Score = 0.61-0.80). Tramadol demonstrated fair agreement (Cohen's Kappa Score = 0.41-0.60). The lowest measure of agreement was observed with cannabinoids, meeting criteria for slight agreement (Cohen's Kappa Score = 0.01-0.20). An application of the techniques described in this study could be implemented in postmortem toxicology laboratories as well as medical examiners offices to provide preliminary drugs of abuse test results that can be used to direct additional testing. This study highlights the successful integration of a novel specimen matrix and an "off-label" use of an established analytical technique.

3.
J Extracell Vesicles ; 13(6): e12467, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38898558

RESUMO

Extracellular vesicles (EVs) carry disease-specific molecular profiles, demonstrating massive potential in biomarker discovery. In this study, we developed an integrated biochip platform, termed EVID-biochip (EVs identification and detection biochip), which integrates in situ electrochemical protein detection with on-chip antifouling-immunomagnetic beads modified with CD81 antibodies and zwitterion molecules, enabling efficient isolation and detection of neuronal EVs. The capability of the EVID-biochip to isolate common EVs and detect neuronal EVs associated with Parkinson's disease in human serum is successfully demonstrated, using the transmembrane protein L1-cell adhesion molecule (L1CAM) as a target biomarker. The EVID-biochip exhibited high efficiency and specificity for the detection of L1CAM with a sensitivity of 1 pg/mL. Based on the validation of 76 human serum samples, for the first time, this study discovered that the level of L1CAM/neuronal EV particles in serum could serve as a reliable indicator to distinguish Parkinson's disease from control groups with AUC = 0.973. EVID-biochip represents a reliable and rapid liquid biopsy platform for the analysis of complex biofluids offering EVs isolation and detection in a single chip, requiring a small sample volume (300 µL) and an assay time of 1.5 h. This approach has the potential to advance the diagnosis and biomarker discovery of various neurological disorders and other diseases.


Assuntos
Biomarcadores , Vesículas Extracelulares , Molécula L1 de Adesão de Célula Nervosa , Doença de Parkinson , Doença de Parkinson/metabolismo , Doença de Parkinson/sangue , Doença de Parkinson/diagnóstico , Humanos , Vesículas Extracelulares/metabolismo , Molécula L1 de Adesão de Célula Nervosa/metabolismo , Biomarcadores/sangue , Masculino , Feminino , Biópsia Líquida/métodos , Idoso , Pessoa de Meia-Idade
4.
Adv Sci (Weinh) ; 11(30): e2403516, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38868948

RESUMO

Metabolic dysfunction-associated steatotic liver disease (MASLD) is hallmarked by hepatic steatosis, cell injury, inflammation, and fibrosis. This study elaborates on a multicellular biochip-based liver sinusoid model to mimic MASLD pathomechanisms and investigate the therapeutic effects of drug candidates lanifibranor and resmetirom. Mouse liver primary hepatocytes, hepatic stellate cells, Kupffer cells, and endothelial cells are seeded in a dual-chamber biocompatible liver-on-a-chip (LoC). The LoC is then perfused with circulating immune cells (CICs). Acetaminophen (APAP) and free fatty acids (FFAs) treatment recapitulate acute drug-induced liver injury and MASLD, respectively. As a benchmark for the LoC, multiplex immunofluorescence on livers from APAP-injected and dietary MASLD-induced mice reveals characteristic changes on parenchymal and immune cell populations. APAP exposure induces cell death in the LoC, and increased inflammatory cytokine levels in the circulating perfusate. Under FFA stimulation, lipid accumulation, cellular damage, inflammatory secretome, and fibrogenesis are increased in the LoC, reflecting MASLD. Both injury conditions potentiate CIC migration from the perfusate to the LoC cellular layers. Lanifibranor prevents the onset of inflammation, while resmetirom decreases lipid accumulation in hepatocytes and increases the generation of FFA metabolites in the LoC. This study demonstrates the LoC potential for functional and molecular evaluation of liver disease drug candidates.


Assuntos
Acetaminofen , Doença Hepática Induzida por Substâncias e Drogas , Modelos Animais de Doenças , Animais , Camundongos , Dispositivos Lab-On-A-Chip , Hepatócitos/metabolismo , Hepatócitos/efeitos dos fármacos , Fígado/patologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado Gorduroso/metabolismo , Camundongos Endogâmicos C57BL , Propionatos , Chalconas
5.
Biosensors (Basel) ; 14(4)2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38667191

RESUMO

Exosomes constitute an emerging biomarker for cancer diagnosis because they carry multiple proteins that reflect the origins of the parent cell. The highly sensitive detection of exosomes is a crucial prerequisite for the diagnosis of cancer. In this study, we report an exosome detection system based on quantum weak value amplification (WVA). The WVA detection system consists of a reflection detection light path and a Zr-ionized biochip. Zr-ionized biochips effectively capture exosomes through the specific interaction between zirconium dioxide and the phosphate groups on the lipid bilayer of exosomes. Aptamer-modified gold nanoparticles (Au NPs) are then used to specifically recognize proteins on exosomes to enhance the detection signal. The sensitivity and resolution of the detection system are 2944.07 nm/RIU and 1.22 × 10-5 RIU, respectively. The concentration of exosomes can be directly quantified by the WVA system, ranging from 105-107 particles/mL with the detection limit of 3 × 104 particles/mL. The use of Au NPs-EpCAM for the specific enhancement of breast cancer MDA-MB-231 exosomes is demonstrated. The results indicate that the WVA detection system can be a promising candidate for the detection of exosomes as tumor markers.


Assuntos
Técnicas Biossensoriais , Neoplasias da Mama , Exossomos , Ouro , Nanopartículas Metálicas , Humanos , Neoplasias da Mama/diagnóstico , Feminino , Ouro/química , Nanopartículas Metálicas/química , Biomarcadores Tumorais , Linhagem Celular Tumoral , Limite de Detecção , Zircônio/química
6.
Electrophoresis ; 45(9-10): 814-828, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38459798

RESUMO

Analysis of short tandem repeats (STRs) is a global standard method for human identification. Insertion/Deletion polymorphisms (DIPs) can be used for biogeographical ancestry inference. Current DNA typing involves a trained forensic worker operating several specialized instruments in a controlled laboratory environment, which takes 6-8 h. We developed the Quick TargSeq 1.0 integrated system (hereinafter abbreviated to Quick TargSeq) for automated generation of STR and DIP profiles from buccal swab samples and blood stains. The system fully integrates the processes of DNA extraction, polymerase chain reaction (PCR) amplification, and electrophoresis separation using microfluidic biochip technology. Internal validation studies were performed using RTyper 21 or DIP 38 chip cartridges with single-source reference samples according to the Scientific Working Group for DNA Analysis Methods guidelines. These results indicated that the Quick TargSeq system can process reference samples and generate STR or DIP profiles in approximately 2 h, and the profiles were concordant with those determined using traditional STR or DIP analysis methods. Thus, reproducible and concordant DNA profiles were obtained from reference samples. Throughout the study, no lane-to-lane or run-to-run contamination was observed. The Quick TargSeq system produced full profiles from buccal swabs with at least eight swipes, dried blood spot cards with two 2-mm disks, or 10 ng of purified DNA. Potential PCR inhibitors (i.e., coffee, smoking tobacco, and chewing tobacco) did not appear to affect the amplification reactions of the instrument. The overall success rate and concordance rate of 153 samples were 94.12% and 93.44%, respectively, which is comparable to other commercially available rapid DNA instruments. A blind test initiated by a DNA expert group showed that the system can correctly produce DNA profiles with 97.29% genotype concordance with standard bench-processing methods, and the profiles can be uploaded into the national DNA database. These results demonstrated that the Quick TargSeq system can rapidly generate reliable DNA profiles in an automated manner and has the potential for use in the field and forensic laboratories.


Assuntos
DNA , Repetições de Microssatélites , Humanos , Repetições de Microssatélites/genética , DNA/análise , DNA/genética , Técnicas de Genotipagem/métodos , Reação em Cadeia da Polimerase/métodos , Genética Forense/métodos , Reprodutibilidade dos Testes , Impressões Digitais de DNA/métodos , Mucosa Bucal/química , Genótipo
7.
J Hazard Mater ; 469: 134037, 2024 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-38521032

RESUMO

Simple yet ultrasensitive and contamination-free quantification of environmental pathogenic bacteria is in high demand. In this study, we present a portable clustered regularly interspaced short palindromic repeats-associated protein 12a (CRISPR/Cas12a) powered Air-displacement enhanced Evanescent wave fluorescence Fiber-embedded microfluidic Biochip (AEFB) for the high-frequency and nucleic acid amplification-free ultrasensitive detection of Escherichia coli O157:H7. The performance of AEFB was dramatically enhanced upon employing a simple air-solution displacement process. Theoretical assays demonstrated that air-solution displacement significantly enhances evanescent wave field intensity on the fiber biosensor surface and increases the V-number in tapered fiber biosensors. Consequently, light-matter interaction is strengthened, and fluorescence coupling and collection efficiency are improved, considerably enhancing sensitivity. By integrating the CRISPR biosensing mechanism, AEFB facilitated rapid, accurate, nucleic acid amplification-free detection of E.coli O157:H7 with polymerase chain reaction (PCR)-level sensitivity (176 cfu/mL). To validate its practicality, AEFB was used to detect E.coli O157:H7 in surface water and wastewater. Comparison with RT-PCR showed a strong linear relationship (R2 = 0.9871), indicating the excellent accuracy and reliability of this technology in real applications. AEFB is highly versatile and can be easily extended to detect other pathogenic bacteria, which will significantly promote the high-frequency assessment and early-warning of bacterial contamination in aquatic environments.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Ácidos Nucleicos , Escherichia coli O157/genética , Sistemas CRISPR-Cas , Reprodutibilidade dos Testes , Microfluídica
8.
Indian J Microbiol ; 64(1): 92-99, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38468745

RESUMO

With the widespread use of rifampicin (RMP) and isoniazid (INH), multidrug resistance (MDR) in Mycobacterium tuberculosis (M.tb) poses a threat to the success of tuberculosis (TB) control programs. We have developed a new polycarbonate track-etched membranes (PC-TEM) based DNA bio-chip designed for rapid detection of mutations conferring MDR in M.tb culture isolates. Bio-chips were designed to contain 14 specific probes for wild type and mutated allele of selected codons within 80 bp rifampicin resistance determining region of rpoB gene, katG gene and mabA-inhA regulatory region. RMP-resistance-associated gene mutation points rpoB 516, 526, 531 and 533, and the INH-resistance-associated gene mutation points katG315 and inhA-15 were targeted. Bio-chip signal was detected using enhanced chemiluminescence. A total of 50 culture isolates that were sensitive or resistant to RMP and/or INH were analyzed by bio-chip. The results of culture-based drug susceptibility testing (DST) were used as the gold standard and gene sequencing was performed to resolve the discordance. Amongst 50 culture isolates, we have detected 18 MDR, 9 RMP mono-resistant, 6 INH mono-resistant, and 17 fully susceptible isolates. The developed DNA bio-chip has a sensitivity of 90% for RMP and MDR and 100% for INH resistance. The bio-chip has a specificity of 100% for RMP and MDR and 88.8% for INH detection. The identification of mutations using the DNA bio-chip was 100% concordant with the sequencing data for the probes covered by the bio-chip. The detection of rpoB, katG and inhA gene mutation points by a DNA bio-chip may be used as a rapid, accurate, and economical, clinical detection method for MDR detection in M.tb. This is very valuable for the control of TB epidemics.

9.
Micromachines (Basel) ; 15(2)2024 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-38398923

RESUMO

In glass molding to produce biochips with micro holes, cavities, and channels, it is important to machine micro molds. This study presents a novel process for fabricating micro pin arrays on carbon graphite, one of the glass molding materials. The micro pin array was used as a mold to fabricate a glass-based micro hole array. Using conventional micro endmill tools, machining micro-cylindrical pins requires complex toolpaths and is time-consuming. In order to machine micro pin arrays with high efficiency, a micro eccentric tool was introduced. Micro pin arrays with a diameter of 200 µm and a height of 200 µm were easily fabricated on graphite using the micro eccentric tool. In the machining of micro pin arrays using eccentric tools, the machining characteristics such as cutting force and tool wear were investigated.

10.
Diagnostics (Basel) ; 14(2)2024 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-38248076

RESUMO

Intracranial chondroid tumors are a heterogeneous group of neoplasms characterized by the presence of a cartilage matrix. These tumors exhibit overlapping clinical and histological features. Mutations in IDH1/2 genes serve as important diagnostic markers of tumor type, particularly chondrosarcoma. To improve the accuracy of IDH1/2 diagnostics, we compared three methods: biochip assay, real-time PCR with DNA melting analysis using TaqMan probes and sequencing (qPCR-DMA-Sanger), and immunohistochemistry (IHC). Tumor samples from 96 patients were investigated. The IDH1 mutations were detected in 34/64 (53%) chondrosarcomas; IHC detected 27/56 (48.2%) mutations, the qPCR-DMA-Sanger method 27/59 (46%) mutations, and the biochip assay revealed 29/60 (48.3%) mutations. The detection of IDH1 mutations in chordoma (2/15) and osteosarcoma (2/7) suggested the need for a revised diagnosis. In benign tumors, IDH1 mutations were present in chondroma (4/6), but absent in chondromyxoid fibroma (0/4). The most frequent IDH1 mutations were R132C (60%), R132L, and R132G (13.5% each), R132H (8%), and R132S (5%). The concordance between the biochip assay and IHC was 90%, between IHC and PCR-DMA-Sanger 83%, and between biochip assay and qPCR-DMA-Sanger was 98%, respectively. No IDH2 mutations were found. The use of independent diagnostic methods may improve the detection of IDH-mutant specimens in chondroid tumors.

11.
Adv Sci (Weinh) ; 11(11): e2306373, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38204202

RESUMO

Detecting pancreatic duct adenocarcinoma (PDAC) in its early stages and predicting late-stage patient prognosis undergoing chemotherapy is challenging. This work shows that the activation of specific oncogenes leads to elevated expression of mRNAs and their corresponding proteins in extracellular vesicles (EVs) circulating in blood. Utilizing an immune lipoplex nanoparticle (ILN) biochip assay, these findings demonstrate that glypican 1 (GPC1) mRNA expression in the exosomes-rich (Exo) EV subpopulation and GPC1 membrane protein (mProtein) expression in the microvesicles-rich (MV) EV subpopulation, particularly the tumor associated microvesicles (tMV), served as a viable biomarker for PDAC. A combined analysis effectively discriminated early-stage PDAC patients from benign pancreatic diseases and healthy donors in sizable clinical from multiple hospitals. Furthermore, among late-stage PDAC patients undergoing chemotherapy, lower GPC1 tMV-mProtein and Exo-mRNA expression before treatment correlated significantly with prolonged overall survival. These findings underscore the potential of vesicular GPC1 expression for early PDAC screenings and chemotherapy prognosis.


Assuntos
Carcinoma Ductal Pancreático , Vesículas Extracelulares , Neoplasias Pancreáticas , Humanos , Biomarcadores Tumorais/genética , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/genética , Vesículas Extracelulares/metabolismo , Glipicanas/genética , Glipicanas/metabolismo , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
12.
Sensors (Basel) ; 23(23)2023 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-38067713

RESUMO

Cell models are one of the most widely used basic models in biological research, and a variety of in vitro cell culture techniques and models have been developed recently to simulate the physiological microenvironment in vivo. However, regardless of the technique or model, cell culture is the most fundamental but crucial component. As a result, we have developed a cell culture monitoring system to assess the functional status of cells within a biochip. This article focuses on a mini-microscope made from a readily available camera for in situ continuous observation of cell growth within a biochip and a pH sensor based on optoelectronic sensing for measuring pH. With the aid of this monitoring system, scientists can keep an eye on cell growth in real time and learn how the pH of the culture medium affects it. This study offers a new approach for tracking cells on biochips and serves as a valuable resource for enhancing cell culture conditions.


Assuntos
Técnicas de Cultura de Células , Microscopia , Técnicas de Cultura de Células/métodos , Concentração de Íons de Hidrogênio
13.
Iran J Pharm Res ; 22(1): e136363, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38116570

RESUMO

Background: Antimicrobial compounds are used in animal husbandry to prevent and treat bacterial diseases and as illegal growth-promoting agents. Due to the excessive and inappropriate use of antibiotics, the antibiotic residues in milk can cause allergic reactions and antibiotic resistance. A rapid biochip-based method for the multi-analyte screening of 6 families of antibiotic residues (quinolones, ceftiofur, florfenicol, streptomycin, tylosin, and tetracyclines) in milk was validated based on Commission Decision 2002/657 and the European guidance for screening methods for veterinary medicinal products. Methods: This methodology allows the 6 antibiotic families to be detected simultaneously, increasing the screening capacity and reducing costs in test settings. The method's applicability was shown by screening 38 UHT cow milk samples taken from Tehran province, IR Iran. Results: The results showed that the positive threshold T was above Fm, and the CCß was below the European Commission's Maximum Residue Limit (MRL) (100 ppb for ceftiofur and tetracycline and 50 ppb for tylosin in milk). Norfloxacin was detected in about 8% of the samples and tylosin in 2.63%. The total antibiotic concentration in UHT cow milk samples was lower than the European Commission's MRL. Conclusions: This study showed that the biochip technique is valid for screening tylosin, ceftiofur, streptomycin, tetracycline, norfloxacin, and florfenicol in milk. It was found that the method was easy, quick, and capable of detecting 6 families of antibiotic residues simultaneously from a single milk sample without sample preparation.

14.
Biomedicines ; 11(10)2023 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-37893142

RESUMO

Progress in macrophage research is crucial for numerous applications in medicine, including cancer and infectious diseases. However, the existing methods to manipulate living macrophages are labor-intense and inconvenient. Here, we show that macrophage membranes can be reconstituted after storage for months at 4 °C, with their CD206 receptor selectivity and specificity being similar to those in the living cells. Then, we have developed a mannose ligand, specific to CD206, linked with PEG as an IR spectroscopy marker to detect binding with the macrophage receptor. PEG was selected due to its unique adsorption band of the C-O-C group at IR spectra, which does not overlap with other biomolecules' spectroscopic feature. Next, competitive binding assay versus the PEG-bound ligand has enabled the selection of other higher-affinity ligands specific to CD206. Furthermore, those higher-affinity ligands were used to differentiate activated macrophages in a patient's bronchoalveolar (BAL) or nasopharyngeal (NPL) lavage. CD206- control cells (HEK293T) showed only non-specific binding. Therefore, biochips based on reconstituted macrophage membranes as well as PEG-trimannoside as an IR spectroscopic marker can be used to develop new methods facilitating macrophage research and macrophage-focused drug discovery.

15.
Biosensors (Basel) ; 13(9)2023 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-37754075

RESUMO

We present a novel and easy approach using a silicon-based impedance chip to determine the concentration of the given aqueous buffer solution. An accurate determination of the post-dilution concentration of the buffers is necessary for ensuring optimal buffer capacity, pH stability, and to assess solution reproducibility. In this study, we focused on phosphate buffer as the test liquid to achieve precise post-dilution concentration determinations. The impedance chip consisting of a top gold ring electrode, where a test volume of 20 µL to 30 µL of phosphate buffer was introduced for impedance measurements within the frequency range of 40 Hz to 1 MHz. For impedance investigation, we used phosphate buffers with three different pH values, and the impedance was measured after diluting the phosphate buffers to a concentration of 1.00 M, 0.75 M, 0.50 M, 0.25 M, 0.10 M, 0.05 M, and 0.01 M. In order to analyze the distinctive changes in the measured impedance, an equivalent circuit was proposed and modeled. From the impedance modeling, we report that the circuit parameter RAu/Si showed exponential dependence on the concentration of phosphate buffer and no dependence on the pH values of the phosphate buffer and on the added volume inside the ring electrode. The proposed silicon-based impedance chip is quick and uses reduced liquid volume for post-dilution concentration measurements of buffers and has perspective applications in the pharmaceutical and biological domains for regulating, monitoring, and quality control of the buffers.


Assuntos
Fosfatos , Silício , Soluções Tampão , Concentração de Íons de Hidrogênio , Impedância Elétrica , Reprodutibilidade dos Testes
16.
Bioengineering (Basel) ; 10(8)2023 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-37627808

RESUMO

Biochips, a novel technology in the field of biomolecular analysis, offer a promising alternative to conventional testing equipment. These chips integrate multiple functions within a single system, providing a compact and efficient solution for various testing needs. For biochips, a pattern-control micro-electrode-dot-array (MEDA) is a new, universally viable design that can replace microchannels and other micro-components. In a Micro Electrode Dot Array (MEDA), each electrode can be programmatically controlled or dynamically grouped, allowing a single chip to fulfill the diverse requirements of different tests. This capability not only enhances flexibility, but also contributes to cost reduction by eliminating the need for multiple specialized chips. In this paper, we present a visible biochip testing system for tracking the entire testing process in real time, and describe our application of the system to detect SARS-CoV-2.

17.
Colloids Surf B Biointerfaces ; 230: 113504, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37597493

RESUMO

In the present study, we developed an amino-polyindole modified phosphorus doped graphitic carbon nitride nanomaterial (APIN/P-g-C3N4) based immunosensing biochip for Serum amyloid A (SAA) biomarker towards early diagnosis of Swine flu. The P-g-C3N4 was synthesis via polycondensation and functionalized with APIN. Further, the biochip was fabricated by modifying the working area of SPE with APIN/P-g-C3N4 using drop cast method, APIN introduced the larger loading of -NH2 group moieties onto P-g-C3N4 matrix and benefitted to reinforced the biomolecules via covalent linkages. The monoclonal anti-SAA was conjugated onto APIN/P-g-C3N4/SPE using EDC-NHS chemistry and BSA was added for non-specific site blocking. The structural, chemical, composition and morphological characteristics of the synthesized, functionalized nanomaterial and fabricated biochips were investigated by XRD, XPS, FT-IR spectroscopy, SEM, FE-SEM and TEM techniques. Further, the electrochemical characterization and response studies of fabricated biochip were analyzed using the CV and DPV techniques. Based on the analytical performance of the proposed immunosensing biochip i.e. BSA/anti-SAA/APIN/P-g-C3N4/SPE, it is capable to detect SAA protein with ultra sensitivity of 79.5 µA log (mL ng-1) cm-2, ultralow limit of detection of 5 ng mL-1 and wider linear detection range of 5 ng mL-1-500 µg mL-1 with quick response time of 10 min. Moreover, the fabricated immunosensing biochips was used to analyse SAA protein in spiked serum samples and the achieved results demonstrated the good agreement with the electrochemical response observed in standard SAA protein samples in analytical solution. The proposed biochip can provide insights for developing a wide range of clinical screening tools for detecting various contagious diseases.


Assuntos
Anticorpos , Grafite , Suínos , Animais , Espectroscopia de Infravermelho com Transformada de Fourier , Fósforo
18.
Traffic Inj Prev ; 24(8): 663-669, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37603111

RESUMO

OBJECTIVE: This study aimed to measure the prevalence of drugs of abuse (DOA) among the goods carriage drivers associated with the southern State of India, Kerala. METHODS: Point-of-collection testing (POCT) of oral fluid collected from the participants (n = 249) was done using the Evidence MultiSTAT DOA Oral Fluid II Assay kits and the Evidence MultiSTAT analyzer. RESULTS: Out of the total samples, 53 (21.29%) were positive for one or more DOA. A high prevalence of tetrahydrocannabinol (THC) (10.04%) and synthetic cannabinoids were detected in the samples. The use of ketamine, alpha-PVP, LSD, methamphetamine, opiate, 6-MAM, benzodiazepines I, methadone, PCP, tramadol, and amphetamine was also detected and their frequency of use ranged between 4.02 and 0.80%. An association between drug abuse and distance of travel was found in drivers in this study, χ2 (5, N = 249) = 123.5, p < 0.001. Confirmatory analysis using ultra-high performance liquid chromatography-tandem mass spectrometry showed excellent agreement with the results of the screening test. CONCLUSIONS: This was the first study conducted among drivers in India for the detection of DOA. Tetrahydrocannabinol (THC) was used more by the goods carriage drivers associated with Kerala State, India. The use of psychoactive substances significantly increased with the distance of travel. Point-of-collection testing (POCT) by the biochip array technology is an efficient method for the detection of these substances.


Assuntos
Dronabinol , Transtornos Relacionados ao Uso de Substâncias , Humanos , Dronabinol/análise , Acidentes de Trânsito , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Metadona/análise , Anfetaminas/análise , Detecção do Abuso de Substâncias/métodos , Saliva/química
19.
Mol Biol (Mosk) ; 57(4): 597-608, 2023.
Artigo em Russo | MEDLINE | ID: mdl-37528780

RESUMO

Ancient DNA analyses help to solve the problems related to the genogeographic origin and migration patterns of populations. The Khazar Khaganate is a subject of controversy among researchers. Its complex historical development, lack of a sufficient number of artistic and written sources, the disappearance of representatives of Khazar culture leaves open the question of the appearance of the Khazars. DNA phenotyping of bone remains from elite burials of the Khazar period of Southern Russia was carried out with respect to eye color, hair color, skin color, and AB0 blood groups. Eight out of 10 individuals had brown eyes, dark hair (to varying degrees), and a predominantly dark skin during their lifetime. Individuals from two burials had gray-blue eyes, and one individual had blond hair. The most probable AB0 blood group was identified in eight people, of which five blood group 0 (I) group, four had blood group A (II), and one had blood group B (III). The allele frequency distribution was assessed for ten population-specific autosomal markers and suggested high heterogeneity for the ethnogeographic origin of the Khazars examined. The results are evidence for ethnocultural, genetic, and phenotypic diversity of the Khazar Khaganate.


Assuntos
Antígenos de Grupos Sanguíneos , Cor de Olho , Humanos , DNA/genética , Sepultamento , Federação Russa
20.
Proc Natl Acad Sci U S A ; 120(33): e2300036120, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37549292

RESUMO

While the world is rapidly transforming into a superaging society, pharmaceutical approaches to treat sarcopenia have hitherto not been successful due to their insufficient efficacy and failure to specifically target skeletal muscle cells (skMCs). Although electrical stimulation (ES) is emerging as an alternative intervention, its efficacy toward treating sarcopenia remains unexplored. In this study, we demonstrate a silver electroceutical technology with the potential to treat sarcopenia. First, we developed a high-throughput ES screening platform that can simultaneously stimulate 15 independent conditions, while utilizing only a small number of human-derived primary aged/young skMCs (hAskMC/hYskMC). The in vitro screening showed that specific ES conditions induced hypertrophy and rejuvenation in hAskMCs, and the optimal ES frequency in hAskMCs was different from that in hYskMCs. When applied to aged mice in vivo, specific ES conditions improved the prevalence and thickness of Type IIA fibers, along with biomechanical attributes, toward a younger skMC phenotype. This study is expected to pave the way toward an electroceutical treatment for sarcopenia with minimal side effects and help realize personalized bioelectronic medicine.


Assuntos
Sarcopenia , Animais , Humanos , Camundongos , Fibras Musculares Esqueléticas , Músculo Esquelético/fisiologia , Fenótipo , Sarcopenia/terapia , Prata
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