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The utilization of 3D cell culture for spheroid formation holds significant implications in cancer research, contributing to a fundamental understanding of the disease and aiding drug development. Conventional methods such as the hanging drop technique and other alternatives encounter limitations due to smaller drop volumes, leading to nutrient starvation and restricted culture duration. In this study, we present a straightforward approach to creating superhydrophobic paper cones capable of accommodating large volumes of culture media drops. These paper cones have sterility, autoclavability, and bacterial repellent properties. Leveraging these attributes, we successfully generate large spheroids of ovarian cancer cells and, as a proof of concept, conduct drug screening to assess the impact of carboplatin. Thus, our method enables the preparation of flexible superhydrophobic surfaces for laboratory applications in an expeditious manner, exemplified here through spheroid formation and drug screening demonstrations. [ABSTRACT FROM AUTHOR]

Paper based lateral flow systems are widely used biosensor platforms to detect biomolecules in a liquid sample. Proteins, bacteria, oligonucleotides, and nanoparticles were investigated in the literature. In this work we designed a magnetic platform including dual magnets and tested the flow of micron size immunomagnetic particles alone and when loaded with cells on two different types of papers. The prewetting conditions of the paper and the applied external magnetic field are the two dominant factors affecting the particle and cell transport in paper. The images recorded with a cell phone, or with a bright field optical microscope were analyzed to measure the flow of particles and cells. The effect of prewetting conditions and magnetic force were measured, and it was shown that in the worst case, minimum 90% of the introduced cells reached to the edge of the paper. The paper based magnetophoretic lateral flow systems can be used for cell assays. [ABSTRACT FROM AUTHOR]

Paper incorporated with ZnO nanostructures (called ZnO paper) is a flexible substrate material with applications in many areas including UV detectors and gas sensors. In this work, we tested the cytotoxicity of ZnO paper against cancer cells. The ZnO paper was placed in culture media of different cancer cell lines including A549, H1299, and WI38 with and without the targeted anti-lung cancer drugs Iressa and Staurosporine, and cell viability was determined. We found that ZnO nanoparticles were cytotoxic to all the cell lines examined, and cell viability was further reduced by increasing the deposition time of these nanoparticles. In addition, cytotoxicity was enhanced when both the ZnO paper and the anticancer drugs were present in the culture media. Our experimental results indicate that ZnO paper can be used for cytotoxicity test and the development of future novel anticancer drugs. [ABSTRACT FROM AUTHOR]

Exosomes are vesicles released by healthy and cancer cells into the extracellular matrix and bodily fluid. Cancer cell-derived exosomes have attracted much attention in early-stage detection and prognostication of treatment response. Thus, detecting exosomes is of great interest to biology and medicine. However, many conventional detection methods require high-cost equipment and centralized laboratory facilities, making diagnostics inaccessible in limited-resource settings. This study reports a proof-of-concept low-cost electrochemical paper-based analytical device to quantify both the total bulk and cancer cell-derived exosomes in cell culture media. The device employs a sandwich immune assay design, where exosomes are initially captured using the electrode-bound generic antibodies (i.e. CD9) and subsequently detected via ovarian cancer-specific CA125 antibodies. Our proposed device quantifies the total bulk exosome concentration with a detection limit of 9.3 × 107 exosomes per mL and ovarian cancer cell-derived exosomes with a detection limit of 7.1 × 108 exosomes per mL, with a relative standard deviation of <10% (n = 3). We suggest that this low-cost and simple electrochemical paper-based device could be an alternative tool for detecting disease-specific exosomes in biological samples with the potential to be further developed for point-of-care diagnosis. [ABSTRACT FROM AUTHOR]

Ovarian cancer has a poor cure rate and rates of relapse are high. Current recurrence detection is limited by non-specific methods such as blood testing and ultrasound. Based on reports that human epididymis four (HE4) / creatinine (CRE) ratios found in urine are elevated in ovarian cancers, we have developed a paper-based device that combines lateral flow technology and cell phone analysis to quantitatively measure HE4/CRE. Surrogate samples were used to test the performance over clinically expected HE4/CRE ratios. For HE4/CRE ratios of 2 to 47, the percent error was found to be 16.0% on average whether measured by a flatbed scanner or cell phone. There was not a significant difference between the results from the cell phone or scanner. Based on published studies, error in this method was less than the difference required to detect recurrence. This promising new tool, with further development, could be used at home or in low-resource settings to provide timely detection of ovarian cancer recurrence. [ABSTRACT FROM AUTHOR]

In this study, a paper-integrated analytical device that combined a paper-based colorimetric assay with a paper-based cell culture platform was developed for the in situ detection of hydrogen sulfide (H2S) in three-dimensional (3D)-cultured, live prostate cancer cells. Two kinds of paper substrates were fabricated using a simple wax-printing methodology to form the cell culture and detection zones, respectively. LNCaP cells were seeded directly on the paper substrate and grown in the paper-integrated analytical device. The cell viability and H2S production of LNCaP cells were assessed using a simple water-soluble tetrazolium salt colorimetric assay and H2S-sensing paper, respectively. The H2S-sensing paper showed good sensitivity (sensitivity: 6.12 blue channel intensity/μM H2S, R2 = 0.994) and a limit of quantification of 1.08 μM. As a result, we successfully measured changes in endogenous H2S production in 3D-cultured, live LNCaP cells within the paper-integrated analytical device while varying the duration of incubation and substrate concentration (L-cysteine). This paper-integrated analytical device can provide a simple and effective method to investigate H2S signaling pathways and drug screening in a 3D culture model. [ABSTRACT FROM AUTHOR]

Highlights • We devised a system of paper-based SERS substrate combined with magnetic separation. • The system was used for simple and effective cancer cell screening. • Distinct SERS spectra of colon cancer cells, normal cells, and RBCs were achieved. • Each cell type could be differentiated with high sensitivity and specificity. • Captured cells could be identified as cancerous cells with accuracy up to 98%. Abstract Early and precise diagnosis of cancer is critical for a better prognosis. Here, we describe a simple and cost-effective plasmonic paper as a surface enhanced Raman scattering (SERS) substrate in combination with magnetic separation for cancer screening. The plasmonic paper was fabricated by immersing plain filter paper into gold nanorod solution and the SERS property of the paper was evaluated using 4-mercaptobenzoic acid (4-MBA) and rhodamine 6 G (R6 G), which showed an enhancement factor (EF) in the range of 106–108. HT-29, a colorectal cancer cell line that highly expresses epithelial cell adhesion molecule (EpCAM), served as the target cells; non-EpCAM-expressing cells, namely fibroblasts and red blood cells (RBCs), were used as negative controls. Intrinsic SERS spectra of the target and control cells showed distinctive patterns on the plasmonic paper due to differences in their structure and components. A combination of principal component analysis (PCA) and k-nearest-neighbor algorithm (k-NN) was employed to analyze and distinguish the acquired HT-29 and fibroblast SERS spectra, demonstrating a diagnostic sensitivity and specificity of 84.8% and 82.6%, respectively, whereas the differentiation between HT-29 and RBCs SERS spectra showed a sensitivity and specificity of 96.4% and 100%, respectively. The magnetic separation was applied to capture the target cells from cell mixtures followed by PCA and k-NN analysis. The identification of the captured cells as cancerous cells from the HT-29 and fibroblast mixture indicated an accuracy of 83.7%, while that from a mixture of HT-29 and RBCs was 98.2%. Thus, the simple paper-based SERS substrate with the assistance of magnetic enrichment and multivariate analysis offers a potent new platform for cancer cell detection and screening. [ABSTRACT FROM AUTHOR]

Highlights • A simple, inexpensive and fully integrated device was demonstrated for detecting gene mutations. • Integration of lateral flow test allowed the detection of gene mutation by naked eyes. • The L858R mutation in lung adenocarcinoma cells was detected with high specificity and sensitivity. Abstract Paper-based diagnostics have been successfully explored for nucleic acid detection. However, multi-step integration (such as nucleic acid extraction, washing, amplification and detection) into a single paper chip remains a huge challenge. Here, a simple, inexpensive, and fully integrated paper chip was demonstrated for detecting gene mutations, which could potentially play a guiding role in individualized treatments to improve the survival rate of lung cancer patients. To simplify the pipetting and manipulation of samples, we incorporated DNA extraction, purification, amplification and detection into a single chip. Further integration of lateral flow test allowed the detection of gene mutation by naked eye. The gene mutation in the epidermal growth factor receptor (EGFR L858R) in lung adenocarcinoma cells was successfully detected with high specificity and sensitivity within 90 min by the proposed fully integrated chip. This low cost, disposable and user-friendly chip provided a new and rapid molecular diagnostic platform, especially suitable for molecular diagnostics in resource-limited areas. [ABSTRACT FROM AUTHOR]

Conventional methods for extraction of DNA are expensive, time-consuming and tedious. To overcome these limitations, a paper-based DNA extraction device was developed that incorporates sponge-based buffer storage, a paper-based valve and channels of different length to autonomously direct the reagent and sample to the Fusion 5 disk for DNA capturing. With this device, DNA can be extracted within 2 min from only 30 μL samples of whole blood, serum, breast cancer cell, saliva, sputum and bacterial suspension. The device can also extract Hepatitis B Virus DNA from clinical blood samples and after quantification shows a detection limit as low as 10 copies⋅mL. This highlights its potential use in future diagnostics. The performance of the device was similar to that of the commercial QIAamp DNA micro kits and the FTA card. In our perception, this simple, inexpensive, portable and disposable device holds great promise in terms of POC testing in resource-poor settings . [ABSTRACT FROM AUTHOR]

We report on the design of a simple all-DNA circuit with dual functions of signal amplification and signal reporting and its use for detection of human telomerase activity from cancer cells. The system utilizes a catalytic hairpin assembly (CHA) reaction for amplification, which produces split G-quadruplex outputs that assemble to form complete guanine quadruplex structures as reporting modules. As designed, a linear DNA sequence (the target) functions as a catalyst to drive cyclic programmed assembly of two hairpins, producing a DNA duplex with two guanine-rich sequences that assemble to form a complete Gq structure. The formation of the Gq element allows either fluorescence or colorimetric detection of the target. Examples are provided to demonstrate fluorescence detection of cancer cells' telomerase activities in solution and the first example of a CHAmodulated colorimetric assay for detecting telomerase activities of cancer cells using a simple paper device. [ABSTRACT FROM AUTHOR]

Digital Signal Processing (DSP) plays an important role in the study of genes and genomics in today's time. DSP is applied to the DNA sequences and then by applying the various DSP algorithms like DFT, (Discrete Fourier Transform), DWT (Discrete Wavelet Transform). We can predict the coding region of DNA and also find out abnormality present in the coding region. This algorithm is used to find out the cancer cells. As cancer is caused due to some mutation or abnormalities present in the DNA. So this algorithm is tested in on the several cancer gene databases and normal gene databases present in the Gene bank and NCBI website that provides satisfactory results. [ABSTRACT FROM AUTHOR]

Abstract: Micrometer‐sized functional nucleic acid (FNA) superstructures (denoted as 3D DNA) were examined as a unique class of biorecognition elements to produce highly functional bioactive paper surfaces. 3D DNA containing repeating sequences of either a DNA aptamer or DNAzyme was created from long‐chain products of rolling circle amplification followed by salt aging. The resulting 3D DNA retained its original spherical shape upon inkjet printing and adhered strongly to the paper surface via physisorption. 3D DNA paper sensors showed resistance to degradation by nucleases, suppressed nonspecific protein adsorption, and provided a much higher surface density of functional DNA relative to monomeric FNAs, making such species ideally suited for development of paper‐based biosensors. [ABSTRACT FROM AUTHOR]

In this paper, a simple but efficient electrochemical biosensing platform was developed for ATP assay based on the preparation of paper-based working electrodes. In contrast to the electrochemical response of commercial glassy carbon electrode (GCE), improved voltammetric signals were achieved at the paper-based electrode. In particular, the surface contamination arising from oxidation product, a prominent challenge for ATP detection, was effectively eliminated by the paper-based system, endowing reliable analysis with high reproducibility. Moreover, the practical application value of the system was verified by assay of ATP in human serums, cancer cells, and normal cells with satisfactory results. Compared with the detection performances of traditional strategies, the advantages involved in the system were exhibited to be rapid, accurate, convenient, and inexpensive, which confer the proposed sensing platform promising for applications in public health as well as the fundamental research of molecular biology. [ABSTRACT FROM AUTHOR]

In this manuscript, a disposable paper-based analytical device comprised of a closed bipolar electrode (BPE) was fabricated for the ultrasensitive electrochemiluminescence (ECL) detection of intracellular H 2 O 2 and the number of cancer cells. In this approach, wax printing was used to fabricated reaction zone, and carbon ink-based BPE and driving electrodes were screen-printed into the paper. AuPd nanoparticles (NPs), which served as a carrier of the capture aptamer and as the catalyst for the ECL reaction of luminol and H 2 O 2 , were used to modify the BPE. Luminol/Au NPs were attached to the surface of the captured cells via hybridation chain reaction with two hairpin structure DNA labelled luminol/Au NPs. In the stimulation of phorbol myristate acetate, The coreactant H 2 O 2 was released from the target cells. The ECL response of the luminol-H 2 O 2 system was related to the number of cancer cells in the testing buffer, which served as a quantitative signal for the determination of cancer cells and the concentration of H 2 O 2 . In order to decrease the external voltage, K 3 [Fe(CN) 6 ] was introduced in the cathode resevoir of BPE because it gained electrons at the cathode more easily than oxygen. The ECL intensity was quantitatively related to the concentration of MCF-7 in the range of 1.0 × 10 2 –1.0 × 10 7 cells/mL. The detection limit was 40 cells/mL and it showed good specificity for cells with high overexpression of mucin-1 receptor, it was concluded that the developed protocol could be effectively utilized for the detection of MCF-7 cells. [ABSTRACT FROM AUTHOR]

The article cites a research study that examines the role of MiR- Par3L axis in breast cancer progression. It examined normal tissues and Par3L expressions in breast cancer tissues and cells. Par3L was found to be frequently down-regulated in human breast cancer cells and tissues and Par3L low-expression was associated with aggressive clinicopathological features and poor clinical outcome of breast cancer patients.

A paper-based portable and disposable cyto-device was fabricated for the purpose of detecting cancer cells and in-situ screening of anticancer drugs. The device was prepared as follows: (a) Au@Pd paper electrodes were prepared by growing Au@Pd nanoparticles on cellulose; (b) the aptamer against MCF-7 cells was immobilized on the paper electrodes; (c) PtNi alloy particles were loaded with carbon dots to form a PtNi@C-dot composite acting as the ECL label, (d) concanavalin A (Con A) was labeled with the ECL label and then dropped on sensor surface carrying cancer cell. Con A can specifically recognize the mannose on the cancer cell surface. The assay is performed in the following way: (1) The aptamer-modified electrode is placed in a solution containing MCF-7 cancer cells; (2) buffer containing peroxodisulfate is added; (3) a potential sweep (from −0.5 to −1.8 V) is applied to the electrode, and ECL intensity is recorded. ECL intensity is related to the number of captured MCF-7 cells in the 480 to 2.0 × 10 cells⋅mL‾ concentration range. The electrode was applied to design a screening method for anti-cancer drugs. This is accomplished by monitoring the number of apoptotic cancer cells after exposing the cells on the electrode to a drug-containing medium. In our perception, the method presented here has a wide range of potential applications in cancer diagnosis and treatment. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

In this work, a novel dual photoelectrochemical/colorimetric cyto-analysis format was first introduced into a microfluidic paper-based analytical device (μ-PAD) for synchronous sensitive and visual detection of H2O2 released from tumor cells based on an in situ hydroxyl radicals (•OH) cleaving DNA approach. The resulted μ-PAD offered an excellent platform for high-performance biosensing applications, which was constructed by a layer-by-layer modification of concanavalin A, graphene quantum dots (GQDs) labeled flower-like Au@Pd alloy nanoparticles (NPs) probe, and tumor cells on the surface of the vertically aligned bamboo like ZnO, which grows on a pyknotic Pt NPs modified paper working electrode (ZnO/Pt-PWE). It was the effective matching of energy levels between GQDs and ZnO levels that lead to the enhancement of the photocurrent response compared with the bare ZnO/Pt-PWE. After releasing H2O2, the DNA strand was cleaved by •OH generated under the synergistic catalysis of GQDs and Au@Pd alloy NPs and thus, reduced the photocurrent, resulting in a high sensitivity to H2O2 in aqueous solutions with a detection limit of 0.05 nmol observed, much lower than that in the previously reported method. The disengaged probe can result in catalytic chromogenic reaction of substrates, resulting in real-time imaging of H2O2 biological processes. Therefore, this work provided a truly low-cost, simple, and disposable μ-PAD for precise and visual detection of cellular H2O2, which had potential utility to cellular biology and pathophysiology. [ABSTRACT FROM AUTHOR]

A trimetallic dendritic Au@PdPt nanparticles (Au@PdPt NPs) with intrinsic peroxidase-like catalytic activity were fabricated by a facile method. Au@PdPt NPs catalyzed the reduction of H 2 O 2 with the help of the electron mediator (thionine), which produced the electrochemical signals dramatically amplified. A sandwiched sensor was designed and implemented in the paper chip device for the determination of K-562 cells. The surface of dendritic Au@PtPd NPs was linked with folic acid by click chemistry. Folic acid was attached to the folate receptor (FR) of cell surface with high affinity. So, K-562 cells could be assayed based on folic acid-functionalized Au@PtPd NPs. On the other hand, folic acid-functionalized Au-paper working electrode on origami electrochemical device was fabricated, which could be used to capture K-562 cells. The twice utilization of folic acid for the capture of K-562 cells as well as the attachment of dendritic Au@PtPd NPs nanoprobe, which enhanced the selectivity of sensor for the determination of cells. The proposed biosensor exhibited excellent analytical performance toward the cytosensing of K562 cells with a wide detection linear range from 1.0 × 10 2 to 2.0 × 10 7 cells mL −1 and a detection limit of 31 cells mL −1 . Further, the proposed electrochemical sensor was applied to monitor cells. Au@PtPd NPs nonenzymatic catalysts could be used as signal-amplifying nanoprobes for ultrasensitive electrochemical sensing. [ABSTRACT FROM AUTHOR]

A novel electrochemical lab-on-paper cyto-device (ELPCD) was fabricated to demonstrate sensitive and specific cancer cell detection as well as in-situ monitoring of multi-glycans on living cancer cells. In this ELPCD, aptamers modified three-dimensional macroporous Au-paper electrode (Au-PE) was employed as the working electrode for specific and efficient cancer cell capture. Using a sandwich format, sensitive and reproducible cell detection was achieved in this ELPCD on the basis of the electrochemical signal amplification of the Au-PE and the horseradish peroxidase-lectin electrochemical probe. The ELPCD displayed excellent analytical performance for the detection of four K562 cells with a wide linear calibration range from 550 to 2.0×10 7 cells mL −1 . Then, this ELPCD was successfully applied to determine cell-surface multi-glycans in parallel and in-situ monitor multi-glycans expression on living cells in response to drug treatment through in-electrode 3D cell culture. The proposed method provides promising application in decipherment of the glycomic codes as well as clinical diagnosis and treatment in early process of cancer. [ABSTRACT FROM AUTHOR]

The detection of cancer cells is important and fundamental for cancer diagnosis and therapy, which has attracted considerable interest recently. Although traditional cyto-sensors have been widely explored due to their high sensitivity and selectivity, it is still a challenge to develop a low-cost, portable, disposable, fast, and easy-to-use cancer cell detection method for applying in the field of cancer diagnosis and therapy. Herein, to address these challenges, we developed a microfluidic paper-based electrochemiluminescence origami cyto-device (μ-PECLOC), in which aptamers modified 3D macroporous Au-paper electrodes were employed as the working electrodes and efficient platforms for the specific cancer cells capture. Owing to the effective disproportionation of hydrogen peroxide and specific recognition of mannose on cell surface, concanavalin-A conjugated porous AuPd alloy nanoparticles were introduced into this μ-PECLOC as the catalytically promoted nanolabels for peroxydisulfate ECL system. Under the optimal conditions, the proposed μ-PECLOC exhibited excellent analytical performance with good stability, reproducibility, and accuracy, towards the cyto-sensing of four types of cancer cells indicating the potential applications to facilitate effective and multiple early cancer diagnosis and clinical treatment. [ABSTRACT FROM AUTHOR]

In this work, using human acute promyelocytic leukemia cells (HL-60) as a model, a novel microfluidic paper-based electrochemical cyto-device (μ-PECD) was fabricated to demonstrate a facile, portable, and disposable approach for cancer cell detection and in situ screening of anticancer drugs in a high-throughput manner. In this μ-PECD, aptamers modified three-dimensional macroporous Au-paper electrode (Au-PE) was fabricated and employed as the working electrode for specific and efficient cancer cell capture as well as for sequential in-electrode 3D cell culture. This Au-PE showed enhanced capture capacity for cancer cells and good biocompatibility for preserving the activity of captured living cells. Sensitive cancer cell detection was achieved in this μ-PECD, which could respond down to four HL-60 cells in 10 μL volume with a wide linear calibration range from 5.0 × 10 2 to 7.5 × 10 7 cells mL −1 and exhibited good stability and reproducibility. Then, in situ anticancer drug screening was successfully implemented in this μ-PECD through monitoring of the apoptotic cancer cells after the in-electrode 3D cell culture with drug-containing culture medium, demonstrating its wide range of potential applications to facilitate effective clinical cancer diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Detection of target analytes with high sensitivity and reproducibility remains a challenge for surface-enhanced Raman scattering (SERS) due to the lack of cost-effective and highly sensitive substrates. In this study, a hydrophobic SERS substrate capable of concentrating nanoparticles and analytes was prepared by spin-coating lubricating liquid onto commercial paper. The condensation effect of the paper-based hydrophobic substrate induced aggregation of gold nanoparticles (Au NPs) to generate "hot spots" for SERS and to drive analytes to the hot-spot areas for more sensitive detection. The obtained SERS signal intensity was 5-fold higher than that obtained using common paper, and a detection limit (LOD) of 4.3 × 10−10 M for rhodamine 6G (R6G) was achieved. Randomly selected points on the substrate and different batches of substrates all exhibited high reproducibility, and the relative standard deviation (RSD) at 1362 cm−1 is approximately 11%. A further application of the hydrophobic substrate was demonstrated by the detection of cytochrome C within a linear detection range of 3.90 × 10−8 M–1.25 × 10−6 M. In addition, the prepared substrate can obtain identifiable SERS spectra of cancer cells and non-cancer cells because a large number of AuNP or Au NPs clusters can adhere to cells, resulting in the construction of a 3D hotspot matrix. The disposable hydrophobic paper substrate eliminates the problem of solution diffusion, and also provides an effective platform for biomolecular screening detection. Image 1 • A hydrophobic substrate capable of highly sensitive SERS detection was prepared. • The molecules could be driven to hot-spot areas of nanoparticles via the solvent evaporation. • The substrate eliminates the problem of solution diffusion. • The substrate provides a non-destructive method for distinguish cancer and non-cancer cells. [ABSTRACT FROM AUTHOR]

A novel paper working electrode with Pt nanospheres grown in it (Pt-PWE) was first used as a sensor platform and then cancer cells were immobilized on the Pt-PWE (high affinity binding with aptamers). This electrode was first designed to achieve the in situ and real-time determination of H2O2 released from cancer cells to obtain an accurate determination. [ABSTRACT FROM AUTHOR]

Water soluble pullulan films were formatted into paper-based microfluidic devices, serving as a controlled time shutoff valve. The utility of the valve was demonstrated by a one-step, fully automatic implementation of a complex pesticide assay requiring timed, sequential exposure of an immobilized enzyme layer to separate liquid streams. Pullulan film dissolution and the capillary wicking of aqueous solutions through the device were measured and modeled providing valve design criteria. The films dissolve mainly by surface erosion, meaning the film thickness mainly controls the shutoff time. This method can also provide time-dependent sequential release of reagents without compromising the simplicity and low cost of paper-based devices. [ABSTRACT FROM AUTHOR]

IOS Press has retracted the following publication from its online content: [Cancer Biomarkers, 24(4) (2019), 497-508, DOI: 10.3233/CBM-182222] B Schizandrin A inhibits proliferation, migration and invasion of thyroid cancer cell line TPC-1 by down regulation of microRNA-429 b Qiuli Ding, Xiaoyan Li, Yongcun Sun and Xinru Zhang I Department of Ultrasound, Jining No.1 People's Hospital, Jining, Shandong 272000, China i This paper belongs to a set of papers allegedly linked to a paper mill, as described in [[1]]. The editorial office contacted the authors to clarify the allegations, but to date there was no response; therefore the article has been retracted. [Extracted from the article]

Celecoxib, a selective cyclooxygenase 2 inhibitor, is known to have anti-inflammatory activity and to induce apoptosis in various types of cancer cells. Here, we examined the molecular mechanism of celecoxib-induced apoptosis in cervical cancer cell lines (HeLa, CaSki and C33A). Screening of a microarray cDNA-chip containing 225 different genes revealed that growth arrest and DNA damage inducible gene (GADD153), a transcription factor involved in apoptosis, showed the strongest differential expression following celecoxib treatment in all three cervical cancer cell lines. Notably, siRNA-induced silencing of GADD153 suppressed celecoxib-induced apoptosis in all the three cell lines, and exogenous expression of GADD153 triggered apoptosis in cervical cancer cells in the absence of other apoptotic stimuli. A luciferase reporter gene assay and mRNA stability tests revealed that expression of GADD153 was regulated at both the transcriptional and post-transcriptional levels following celecoxib treatment. The region between −649 and −249, containing an intact C/EBP-ATF binding site, was required for the basal activity and celecoxib-induced stimulation of GADD153 promoter activity. Also, mRNA stability test showed that celecoxib prolonged the half-life of GADD153 mRNA. In terms of signaling pathway, addition of the NF-κB inhibitor, N-tosyl-l phenylalanyl-chloromethyl ketone (TPCK), had no effect on GADD153 expression levels. Celecoxib treatment induced Bak expression, whereas cell treated with siGADD153 or TPCK showed lower levels of celecoxib-induced Bak up-regulation. These novel findings collectively suggest that GADD153 may play a key role in celecoxib-induced apoptosis in cervical cancer cells by regulating the expression of proapoptotic proteins such as Bak. [ABSTRACT FROM AUTHOR]

The article reports on several research papers related to cancer and its treatment. It informs that loss of the apoptotic response has been implicated in carcinogenesis as a consequence of increased survival of DNA-damage bearing cells. Many cancer cells have an inherent resistance to chemotherapy due to the abnormal expression of anti-apoptosis genes. The increased risk of cancer in immunosuppressed patients after organ transplantation, and in patients with inherited defects in immune function, such as severe combined immune deficiency, was thought to support the immune surveillance hypothesis.

• We introduce a colorimetric and SERS sensor for the dual-mode detection of H 2 S. • A simple and facile H 2 S assay was prepared by dropping of Ag nanoplate solution on paper. • We utilized the parameter b of CIE L*a*b color model for the quantitative color analysis. • The analytical performance of H 2 S sensing paper for dual-mode detection was evaluated. • This paper assay can detect endogenous H 2 S in live LNCaP cells under shorter incubation times. We report a simple and facile paper assay based on silver nanoplates (Ag NPls) for the colorimetric and surface-enhanced Raman scattering (SERS) dual-mode detection of hydrogen sulfide (H 2 S) gas released from live cancer cells. Ag NPls were prepared via seed-mediated growth and dropped on detection zones within the poly(dimethylsiloxane)-patterned paper substrate. The blue-colored Ag NPls on the paper react with H 2 S to form brown-colored silver sulfide, which simultaneously decreases SERS activity of Ag NPls. We quantitatively analyzed the color and Raman signal intensity changes in Ag NPl-coated detection zone on the paper, using the parameter b of L*a*b color space and Raman peak at 1356 cm−1 of Rhodamine B, respectively. As a result, our Ag NPl-coated H 2 S dual-sensing paper assay showed a good sensitivity with a linear range of 1.64–16.4 μM H 2 S and a detection limit of 520 nM for colorimetric detection. And, it also represented a high sensitivity with a linear range of 82–330 nM H 2 S for SERS detection. In addition, it possessed high selectivity, excellent reproducibility, and high stability over 1 month. Using this Ag NPl-coated H 2 S dual sensing paper, we successfully measured the H 2 S concentration in live prostate cancer cells at different incubation times and cell numbers. Therefore, the Ag NPl-coated paper assay can be utilized as a facile and effective tool for the dual-mode colorimetric and SERS detection of free H 2 S gas within live cells without complicated sample processing or labor-intensive bioanalytical methods. Image, graphical abstract [ABSTRACT FROM AUTHOR]

In this paper, we present Raman imaging as a non-invasive approach for studying changes in mitochondrial metabolism caused by cardiolipin–cytochrome c interactions. We investigated the effect of mitochondrial dysregulation on cardiolipin (CL) and cytochrome c (Cyt c) interactions for a brain cancer cell line (U-87 MG). Mitochondrial metabolism was monitored by checking the intensities of the Raman bands at 750 cm−1, 1126 cm−1, 1310 cm−1, 1337 cm−1, 1444 cm−1 and 1584 cm−1. The presented results indicate that under pathological conditions, the content and redox status of Cyt c in mitochondria can be used as a Raman marker to characterize changes in cellular metabolism. This work provides evidence that cardiolipin–cytochrome c interactions are crucial for mitochondrial energy homeostasis by controlling the redox status of Cyt c in the electron transport chain, switching from disabling Cyt c reduction and enabling peroxidase activity. This paper provides experimental support for the hypothesis of how cardiolipin–cytochrome c interactions regulate electron transfer in the respiratory chain, apoptosis and mROS production in mitochondria. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are considered as the potential biomarkers for many cancers. To determine miRNAs in cancer cells is significant for realizing these diseases. In this work, a microfluidic paper-based laser-induced fluorescence sensor based on duplex-specific nuclease (DSN) amplification was developed and applied to selectively and sensitively determine miRNAs in cancer cells. An interface for laser-induced fluorescence detection was firstly applied to perform the sample detection on the paper-based chip. Under the optimal conditions, DSN (3 μL 0.10 U) and Taqman probes (2 μL 2.5 × 10−7 M) were preserved on the circles (Diameter 4 mm) of the folded paper chip. When miRNA solution was added, the mixed solution could trigger fluorescence signal amplification by cyclically digesting hybrids of miRNAs and Taqman probes by DSN. The whole determination, including sample heating process, could be accomplished within 40 min. The detection limits for miRNA-21 and miRNA-31 were 0.20 and 0.50 fM respectively, corresponding to only 1.0 and 1.5 zmol consumption of miRNAs. The testing of mismatched miRNAs showed that the method had good specificity. Finally, the method was applied to determine miRNA-21 and miRNA-31 in lysates of cancer cells of A549 and HeLa, and hepatocyte LO2. MiRNA-21 and miRNA-31 could be successfully found from the two cancer cells. The concentrations for miRNA-21 and miRNA-31 were 1.74 × 10−13 M and 6.29 × 10−14 M in HeLa cell lysate (3.75 × 104 cells/mL), 3.07 × 10−15 M and 3.28 × 10−15 M in A549 cell lysate (8.33 × 106 cells/mL) respectively. The recoveries ranged from 87.30% to 111.83%, indicating the results were reliable. The developed method was effective, selective and sensitive in the determination of miRNAs in cancer cells. Image 1 • Realization of sensitive laser-induced fluorescence detection with an interface. • Performing duplex-specific nuclease amplification on the paper-based chip. • The applicable method determined low abundance miRNAs from cancer cells. [ABSTRACT FROM AUTHOR]

Abstract: Rapid signaling of estrogen involves membrane estrogen receptors (ERs), including membrane subpopulations of ERα and ERβ. In the mid-1990s, several laboratories independently reported the cloning of an orphan G protein-coupled receptor from vascular and cancer cells that was named GPR30. Research published between 2000 and 2005 provided evidence that GPR30 binds and signals via estrogen indicating that this intracellular receptor is involved in rapid, non-genomic estrogen signaling. The receptor has since been designated as the G protein-coupled estrogen receptor (GPER) by the International Union of Pharmacology. The availability of genetic tools such as different lines of GPER knock-out mice, as well as GPER-selective agonists and antagonists has advanced our understanding, but also added some confusion about the new function of this receptor. GPER not only binds estrogens but also other substances, including SERMs, SERDs, and environmental ER activators (endocrine disruptors; xenoestrogens) and also interacts with other proteins. This article represents a summary of a lecture given at the 7th International Meeting on Rapid Responses to Steroid Hormones in September 2011 in Axos, Crete, and reviews the current knowledge and questions about GPER-dependent signaling and function. Controversies that have complicated our understanding of GPER, including interactions with human ERα-36 and aldosterone as a potential ligand, will also be discussed. [Copyright &y& Elsevier]

• Developing a mini paper-based device to detect fluorene cytotoxicity in 10 µL sample. • The MWCNTs/μ-PAD allowed a sampling volume of 10.0 μL, which is 50.0 times less than the sampling volume required with a conventional electrode (500 μL). • This device is comparable to the MTT method in fluorene cytotoxicity analysis. Developing a cost-effective and simple micro-analysis tool has long been an important objective in the toxicological detection of fluorene. In this study, a disposable paper-based micro-analysis device (μ-PAD) was designed using graphite doped with multi-walled carbon nanotubes (MWCNTs) to hand draw the electrode (MWCNTs/μ-PAD). We investigated the feasibility of the designed MWCNTs/μ-PAD as a cell-sensing platform using voltammetry measurements. Its application for evaluating the cytotoxicity of fluorene was studied based on the electrochemical determination of human breast cancer cells induced by fluorene. The results showed a dose-dependent toxicity effect of fluorene on cellular activity. The measurements were comparable with those obtained using a methyl-thiazolyl-tetrazolium assay. The detection limit of the MWCNTs/μ-PAD for human breast cancer cells was as low as 4.00 × 103 cells·mL−1 owing to the enhanced catalytic activity of the MWCNTs. Notably, the MWCNTs/μ-PAD—which had a diameter of 7.00 mm—allowed a sampling volume of 10.0 μL, which is 50.0 times less than the sampling volume required with a conventional electrode (500 μL). Advantages such as the simplicity of manufacture, low consumption, low cost, rapid detection, and disposability, suggest that the MWCNTs/μ-PAD could provide new opportunities and directions for in vitro microanalysis. [ABSTRACT FROM AUTHOR]

The interactions between Hela cells and silver nanoparticles (AgNPs) have been studied by scanning electrochemical microscopy (SECM) with both IrCl62−/3−and Fe(CN)63−/4−as the dual mediators. IrCl62−, which can be produced in situand react with AgNPs, is used as the mediator between the AgNPs on the cells and the SECM tip. Another redox couple, Fe(CN)63−/4−, which has a similar hydrophilicity to IrCl62−/3−, but cannot react with AgNPs, is also employed for the contrast experiments. The cell array is cultured successfully onto a Petri dish by microcontact printing (μCP) technique, which can provide a basic platform for studying of single cells. The approach curve and line scan are the two methods of SECM employed here to study the Hela cells. The former can provide the information about the interaction between Hela cells and AgNPs whereas the later gives the cell imaging. The permeability of cell membranes and morphology are two main factors which have effects on the feedback mode signals when K3Fe(CN)6is used as the mediator. The permeability of the cell membranes can be ignored after interaction with high concentration of AgNP solution and the height of the Hela cells is slightly decreased in this process. The kinetic rate constants (k0) between IrCl62−and Ag on the Hela cell can be evaluated using K3IrCl6as the mediator, and they are increased with the higher concentrations of the AgNP solutions. The k0is changed about 10 times from 0.43 ± 0.04 × 10−4to 1.25 ± 0.07 × 10−4and to 3.93 ± 1.9 × 10−4cm s−1corresponding to 0, 1 and 5 mM of AgNO3solution. The experimental results demonstrate that the AgNPs can be adsorbed on the cell surface and detected by SECM. Thus, the amount of AgNPs adsorbed on cell membranes and the permeability or morphology changes can be investigated simultaneously using this approach. The dual mediator system and cell array fabricated by μCP technique can provide better reproducibility because they can simplify experiments, and provide a platform for further single cell detection. [ABSTRACT FROM AUTHOR]

Multiple myeloma (MM) cells express certain tumour-associated antigens (TAAs) that could serve as targets for active-specific immunotherapy. The aim of the present study was to test the MM/dendritic cell (DC) fusion as a vaccination strategy. We fused MM cells with DC to generate fusion cells (FCs) and tested their antigen presenting cell (APC) function in mixed lymphocyte reactions and cytotoxicity assays. First, the HS Sultan and SK0-007 HAT sensitive human MM cell lines and DCs generated from peripheral blood of normal donors were fused in the presence of 50% polyethylene glycol to form FCs. Next, tumour cells freshly isolated from patients were similarly fused with autologous DCs to generate FCs. The FCs demonstrated a biphenotypic profile, confirmed both by flow-cytometry and dual immunofluorescence microscopy. These FCs induced MM-specific cytotoxicity. FCs, but not MM cells or DCs alone, were potent stimulators of autologous patient T cells. More importantly, FC-primed autologous peripheral blood mononuclear cells demonstrated major histocompatibility complex-restricted MM-specific cytolysis. These studies therefore demonstrated that MM/DC FC can trigger an autologous immune response to MM cells and formed the framework for a clinical trial currently underway. [ABSTRACT FROM AUTHOR]

Multiple myeloma (MM) is a neoplastic proliferation of plasma cells and remains an incurable disease because of the development of drug resistance. Histone deacytylase (HDAC) inhibitors are a new class of chemotherapeutic reagents that cause growth arrest and apoptosis of neoplastic cells. Depsipeptide, a new member of the HDAC inhibitors, was found to be safe in humans and has been shown to induce apoptosis in various cancers. In order to evaluate the effects of depsipeptide, a MM cell line, U266 [interleukin (IL)-6 dependent], was analysed for viability and apoptosis. The combined effect of depsipeptide with melphalan and changes in BCL-2 family proteins (BCL-2, BCL-XL, BAX and MCL-1) were also investigated. In addition, the RPMI 8226 cell line (IL-6 independent), and primary patient myeloma cells were also analysed for apoptosis after depsipeptide treatment. Depsipeptide induced apoptosis in both U266 and RPMI 8226 cell lines in a time- and dose-dependent fashion, and in primary patient myeloma cells. We also demonstrated that depsipeptide had an additive effect with melphalan (10 μmol/l). BCL-2, BCL-XL and MCL-1 showed decreased expression in depsipeptide-treated samples. Based on recent clinical trials demonstrating minimal clinical toxicity, our study supports the future clinical utilization of depsipeptide in the management of MM. [ABSTRACT FROM AUTHOR]

The role of repopulating lymphocytes after allogeneic stem cell transplantation (SCT) includes the prevention of serious infections and attacking residual tumour cells in the early post-transplant phase. Therefore, the current study analysed the role of the absolute lymphocyte count (ALC) on day 21 after SCT in predicting transplant outcomes of 82 patients in terms of the risk of opportunistic infections and recurrence of original disease. The median dose of CD34+, CD3+ and mononuclear cells (MNC) infused was 6·41 × 106/kg, 1·96 × 108/kg and 6·81 × 108/kg respectively. The high ALC group (high ALC on day 21; ≥0·35 × 109/l) was associated with the use of peripheral blood stem cells, matched sibling donors and higher cell doses of MNC, CD3+ and CD4+ cells. The high ALC group also exhibited a better overall survival (56·3% vs. 17·7%) and disease-free survival (50·1% vs. 15·9%) after 3 years and lower incidences of relapse (33·6% vs. 67·1%) and fungal infections (3·0% vs. 19·5%) after 1 year. The incidence of cytomegalovirus antigenaemia was lower in the high ALC group (47·7% vs. 73·7%). Accordingly, identifying the ALC on day 21 would appear to be a useful and simple measurement to predict those patients with a high risk of opportunistic infections and relapse after allogeneic SCT. [ABSTRACT FROM AUTHOR]

Bispecific antibodies offer the possibility of improving effector-cell recruitment for antibody therapy. For this purpose, a recombinant bispecific single-chain Fv antibody (bsscFv), directed against Fc γRIII (CD16) and human leucocyte antigen (HLA) class II, was constructed and tested in functional assays. RNA from the hybridomas 3G8 and F3.3, reacting with CD16 and HLA class II, respectively, was used to generate phage display libraries. From these libraries, reactive phages were isolated and the bsscFv was constructed by connecting both single-chain Fv components through a 20 amino acid flexible linker. After expression in SF21 insect cells and chromatographic purification, the bsscFv bound specifically and simultaneously to both antigens. The affinities of the anti-CD16 and the anti-HLA class II scFv components of the bsscFv were 8·6 × 10−8 mol/l and 13·7 × 10−8 mol/l, respectively, which was approximately sevenfold lower than the F(ab) fragments of the parental antibodies. In antibody-dependent cellular cytotoxicity experiments with human mononuclear cells as effectors, the bsscFv-mediated specific lysis of both HLA class II-positive, malignant human B-lymphoid cell lines and primary cells from patients with chronic B-cell lymphocytic leukaemia. Optimal lysis was obtained at bsscFv concentrations of approximately 400 ng/ml, similar to the concentration required for maximum lysis by the corresponding chemically linked bispecific antibody. Thus, this recombinant bsscFv-antibody is an efficient molecule for effector-cell mediated lysis of malignant human B-lymphoid cells. [ABSTRACT FROM AUTHOR]

To evaluate the clinical significance of CD21S expression of diffuse large B-cell lymphoma (DLBCL) tumour cells, we compared their clinical features, immunophenotype, response to therapy and outcome in relation to CD21S expression. Between 1987 and 1999, frozen sections from 240 DLBCL cases were examined for CD21S expression by immunohistochemical methods. CD21S expression was detected on the tumour cells of 87 (36%) cases. The median age of the CD21S+ DLBCL cases was 65 years (range: 17–84 years), the male–female ratio was 42:45, and they showed the following clinical features: Eastern Cooperative Oncology Group score >1 in 14%, lactate dehydrogenase greater than normal levels in 38%, extranodal sites >1 in 14%, stages III/IV disease at diagnosis in 29%, B symptoms in 17%, and a high/high–intermediate International Prognostic Index (IPI) in 23%. They also showed a better overall survival ( P = 0·00001, log-rank test) and a better complete remission rate ( P = 0·00004, chi-square test) than CD21S− DLBCL. Moreover, CD21S+ DLBCL showed a better survival than CD21S− DLBCL for both low/low–intermediate and high/high–intermediate risk categories of IPI ( P = 0·045 and P = 0·0016 respectively). Multivariate analysis identified CD21S expression as an independent factor for survival when compared with the five IPI factors. These findings indicate that CD21S expression of DLBCL tumour cells is a useful prognostic factor for survival. [ABSTRACT FROM AUTHOR]

Minor histocompatibility antigens (mHAs) are major histocompatibility complex (MHC)-associated peptides, which trigger T-cell responses that mediate graft versus host disease (GVHD) and graft versus leukaemia effects. We recently identified a new mHA epitope, termed ACC-1, which is presented by HLA-A*2402 and encoded by BCL2A1, whose expression is restricted to haematopoietic cells including leukaemic cells. HLA-A24/ACC-1 tetramer detected the presence of ACC-1-specific CD8+ cells in the peripheral blood of a patient up to 7 months following transplantation, and these tetramer-positive cells were expandable in vitro by ACC-1 peptide stimulation. A retrospective analysis of 320 patients with HLA-A*2402 who had received a human leucocyte antigen (HLA) genotypically matched unrelated donor through the Japan Marrow Donor Programme was conducted to determine whether ACC-1 disparity is associated with adverse clinical outcomes such as GVHD. Among these patients, ACC-1 disparity was detected in 55 (17·2%) donor/recipient pairs. After adjusting for known risk factors, the hazard ratios or odds ratios of acute and chronic GVHD, relapse and disease-free survival were not statistically different between patients receiving ACC-1 compatible and incompatible transplantation. These data suggest that disparity of haematopoietic cell-specific mHA, ACC-1, is unlikely at least to augment GVHD, and that T cells specific for ACC-1 may also be used for immunotherapy of recurring leukaemia without GVHD. [ABSTRACT FROM AUTHOR]

Diagnosis of hematological cancer requires complete white blood cell count, followed by flow cytometry with multiple markers, and cytology. It requires substantial time and specialized training. A dual-layer paper microfluidic chip was developed as a quicker, low-cost, and field-deployable alternative to detect ROR1+ (receptor tyrosine-like orphan receptor one) cancer cells from the undiluted and untreated buffy coat blood samples. The first capture layer consisted of a GF/D glass fiber substrate, preloaded with cancer specific anti-ROR1 conjugated fluorescent particles to its center for cancer cell capture and direct smartphone fluorescence imaging. The second flow layer was comprised of a grade 1 cellulose chromatography paper with wax-printed four channels for wicking and capillary flow-based detection. The flow velocity was used as measure of antigen concentration in the buffy coat sample. In this manner, intact cells and their antigens were separated and independently analyzed by both imaging and flow velocity analyses. A custom-made smartphone-based fluorescence microscope and automated image processing and particle counter software were developed to enumerate particles on paper, with the limit of detection of 1 cell/μL. Flow velocity analysis showed even greater sensitivity, with the limit of detection of 0.1 cells/μL in the first 6 s of assay. Comparison with capillary flow model revealed great alignment with experimental data and greater correlation to viscosity than interfacial tension. Our proposed device is able to capture and on-chip image ROR1+ cancer cells within a complex sample matrix (buffy coat) while simultaneously quantifying cell concentration in a point-of-care manner. • Dual layer paper microfluidic chip identifies ROR1+ cells directly from buffy coat blood samples. • Cells are captured on the capture layer, while antigens flow through the flow layer. • Smartphone quantified the cells by microscopic counting and the antigens by flow velocities. • Capillary flow model identified viscosity as a major contributor but not interfacial tension. • Detection limit is 0.1 cells/μL from 10 μL sample, without any dilution, concentration, or incubation. [ABSTRACT FROM AUTHOR]

Introduction and aim. Melanoma is a malignant neoplasm of melanocytic origin, the development of which largely depends on the influence of UV radiation. Its development is preceded by the appearance of a melanocytic (dysplastic) nevus. Treatment of patients with metastases to distant organs remains the greatest challenge, as the results of treatment in patients with stage IV melanoma are still unsatisfactory. The progress in systemic therapy of generalized melanoma is based on two mechanisms: molecularly targeted treatment and non-specific melanoma immunotherapy, which is characterized by a unique action - instead of inducing the destruction of tumor cells, it strengthens or stimulates the anti-tumor response of the patient's immune system. Immunotherapy uses ipilimumab and Nivolumab that uses the natural ability of the immune system to eliminate primary cancer cells. It inhibits the binding of the cytotoxic T-4 antigen to its ligands, thereby enhancing T-cell response and tumor resistance. The aim of the study was a presentation of the role of monoclonal antibodies in the treatment of melanoma. Material and methods. During the preparation of the presentation 4 databases (PubMed, Google Scholar, Sciencedirect, Medscape) were searched for studies science publications from march 2019 to March 2023 in English. The search terms were: „ipilimumab in melanoma" „ monoclonal antibodies in melanoma treatment" "nivolumab in melanoma" in the title or abstract or as keywords. A total of 660 references, excluding duplicates, were identified. Analysis of literature. Studies show that nivolumab in combination with ipilimumab reduced the risk of death by 45%, compared to ipilimumab monotherapy. The two-year overall survival rate was 64% for nivolumab in combination with ipilimumab, 59% for nivolumab monotherapy, and 45% for ipilimumab monotherapy. Conclusion. Monoclonal antibodies against various immune checkpoints have revolutionized the treatment of metastatic and inoperable melanoma. The combination of two monoclonal antibodies, e.g. ipilimumab with nivolumab, is much more effective than the use of these antibodies in monotherapy. [ABSTRACT FROM AUTHOR]