Serodiagnosis of multiple cancers using an extracellular protein kinase A autoantibody-based lateral flow platform.

Extracellular protein kinase A autoantibody (ECPKA-AutoAb) has been suggested as a universal cancer biomarker due to its higher amounts in serum of several types of cancer patients than that of normal individuals. Herein, we first developed a lateral flow immunoassay (LFIA) tool, using a sandwich format, toward ECPKA-AutoAb in human serum. For this format, 3G2 as a capture antibody was identified using hybridoma technique and a series of screenings where it showed superior capacity to recognize Enzo PKA catalytic subunit alpha (Cα), compared to other PKA antibodies and antigens. Using these components, we performed sandwich ELISA toward a mimic and real sample of ECPKA-AutoAb. As per the results, limit of detection (LOD) was found to be 135 ng/mL and ECPKA-AutoAb levels were higher in various cancer patients than in normal individuals like previous studies. Based on these results, we applied this sandwich format into LFIA tool and found that the LOD of the fabricated LFIA tool showed about 3.8 ng/mL using spiked PKA-Ab, which is significantly improved compared to the LOD of sandwich ELISA. Also, the developed LFIA tool demonstrated a remarkable ability to detect significant differences in ECPKA-AutoAb levels between normal and cancer patients within 15 min, showing a potential for point-of-care (PoC) detection. One interesting point is that our LFIA strip contains an additional conjugation pad II, named because of its position behind the conjugation pad, in which PKA Cα is dried, enabling a sandwich format.

Machine Learning-Based Approach to Developing Potent EGFR Inhibitors for Breast Cancer-Design, Synthesis, and In Vitro Evaluation.

The epidermal growth factor receptor (EGFR) is vital for regulating cellular functions, including cell division, migration, survival, apoptosis, angiogenesis, and cancer. EGFR overexpression is an ideal target for anticancer drug development as it is absent from normal tissues, marking it as tumor-specific. Unfortunately, the development of medication resistance limits the therapeutic efficacy of the currently approved EGFR inhibitors, indicating the need for further development. Herein, a machine learning-based application that predicts the bioactivity of novel EGFR inhibitors is presented. Clustering of the EGFR small-molecule inhibitor (∼9000 compounds) library showed that N-substituted quinazolin-4-amine-based compounds made up the largest cluster of EGFR inhibitors (∼2500 compounds). Taking advantage of this finding, rational drug design was used to design a novel series of 4-anilinoquinazoline-based EGFR inhibitors, which were first tested by the developed artificial intelligence application, and only the compounds which were predicted to be active were then chosen to be synthesized. This led to the synthesis of 18 novel compounds, which were subsequently evaluated for cytotoxicity and EGFR inhibitory activity. Among the tested compounds, compound 9 demonstrated the most potent antiproliferative activity, with 2.50 and 1.96 µM activity over MCF-7 and MDA-MB-231 cancer cell lines, respectively. Moreover, compound 9 displayed an EGFR inhibitory activity of 2.53 nM and promising apoptotic results, marking it a potential candidate for breast cancer therapy.

Metals as toxicants in event-based expedited production of children's jewelry.

Globally, the hazardous substance in children's goods is of great concern. Toxic chemicals are potentially harmful to the health and growth of infants and children. Lead (Pb) and cadmium (Cd)-contaminated children's jewelry is widely encountered in many countries. This study aims to determine the concentration of metal toxicants (Pb, Cd, Ni, Cu, Zn, Co, and Fe) in event-based (Independence Day festival) children's jewelry, considering time-limited and fast production products that may compromise the quality and safety parameters during manufacturing. The determinations are for the time-constraint industrial production of children's jewelry in the context of the toxic substances in a variety of base materials used. This is the first time event-based children's jewelry has been monitored and critically assessed for metal contamination. Forty-two samples, including metallic, wooden, textile, rubber, plastic, and paint-coated plastic children's jewelry, were tested. Seventy-four percent of samples detected Pb and Cd in quantifiable amounts. Ni in 71%, Cu in 67%, Co in 43%, and Zn and Fe were detected in 100% samples with quantifiable amounts. Twenty-two ID-CJ samples exceeded the US regulatory limit for Pb and four samples for Cd. However, twenty-nine samples for Pb, eleven for Cd, five for Co, and one for Cu exceeded the EU regulatory limit. The highest concentration of Pb was found in paint-coated plastic jewelry, and the highest Cd was found in metallic jewelry. These results suggest that the potential hazards of event-based children's jewelry deserve the attention of government agencies seeking to limit children's exposure to toxic chemicals. Intergovernmental organizations and individual countries regulate chemicals in consumer products, but a coordinated international approach is lacking. Some continents and countries still lack in regulations for children's products, especially jewelry, and toys.

Development of new TAK-285 derivatives as potent EGFR/HER2 inhibitors possessing antiproliferative effects against 22RV1 and PC3 prostate carcinoma cell lines.

Epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) protein tyrosine kinases co-expressed in various cancers such as ovarian, breast, colon, and prostate subtypes. Herein, new TAK-285 derivatives (9a-h) were synthesised, characterised, and biologically evaluated as dual EGFR/HER2 inhibitors. Compound 9f exhibited IC50 values of 2.3 nM over EGFR and 234 nM over HER2, which is 38-fold of staurosporine and 10-fold of TAK-285 over EGFR. Compound 9f also showed high selectivity profile when tested over a small kinase panel. Compounds 9a-h showed IC50 values in the range of 1.0-7.3 nM and 0.8-2.8 nM against PC3 and 22RV1 prostate carcinoma cell lines, respectively. Cell cycle analysis, apoptotic induction, molecular docking, dynamics, and MM-GBSA studies confirmed the plausible mechanism(s) of compound 9f as a potent EGFR/HER2 dual inhibitor with an effective antiproliferative action against prostate carcinoma.

Construction of a Pentacyclic Framework Enabled by Nickel Catalysis.

We present a novel nickel-catalyzed reaction of indole-tethered 2-alkynylphenol esters with various (hetero)aryl boronic acids, resulting in the synthesis of diversely functionalized pentacyclic benzofurocyclohepta[b]indole derivatives. This unprecedented cascade reaction involves the arylative cyclization of alkynes, nucleophilic attack of the indole moiety on the oxonium ion intermediate, 1,2-alkyl group migration, and aromatization. The synthesized molecules exhibit exceptional cytotoxicity against multiple cancer cell lines while maintaining biocompatibility toward healthy cells.

Contamination by hazardous elements in low-priced children's plastic toys bought on the local markets of Karachi, Pakistan.

Children's plastic toys may contain toxic metals to which infants and young children can be orally exposed and may pose acute or chronic adverse health effects. This research aims to evaluate the total metal concentrations (TMCs) of Pb, Cd, Cr, Ni, Zn, Cu, and Mn in children's plastic toys bought in the local markets of Karachi, Pakistan, and compare TMCs to different toy safety regulatory limits. A total of 44 children's plastic toys sourced in the Karachi local markets were analyzed by an atomic absorption spectrophotometer for contamination of hazardous elements. Toy samples were divided into two groups: plastic toys (DCT) and plastic toys with paints or coatings (DPCT). For plastic toys, 83% (19) of samples had TMCs that exceeded European Union (EU) toy safety regulation limits for Pb, and 65% (15) of samples that exceeded for Cd. For plastic toys with paints or coating, 43% (9) of samples had TMCs that exceeded EU migration limits for Pb and 24% (5) for Cd. More than 20 samples exceeded the United States Consumer Product Safety Commission (US CPSC), Canadian, and Bureau of Indian Standards (BIS) toy safety regulation limits. In toy samples (n = 44), very high TMCs of Pb (64%), Cd (45%), Cr (5%), and Ni (2%) were observed. Zn, Cu, and Mn TMCs existed but were below the regulation limits. The contamination levels of Pb, Cd, Cr, and Ni and smaller extent of Zn, Cu, and Mn still pose health issues in children and may cause serious problems in their health.

Electrochemical detection of caspase-3 based on a chemically modified M13 phage virus.

Caspase-3, a cysteine-dependent protease, is considered a reliable molecular biomarker for the diagnosis and prognosis of apoptosis-related diseases. In this study, we demonstrated a phage-based electrochemical biosensor for the evaluation of cell apoptosis by the sensitive and specific detection of caspase-3. Specifically, for screening of affinity peptide-displayed phages, phage display was performed using M13 phage libraries (cyclic forms of peptides), and we identified potential affinity peptide-displayed phage clones with the sequence CPTTMWRYC. After characterization of its binding affinity using enzyme-linked immunosorbent assay, whole phage particles were covalently attached to a gold surface using coupling chemistry (MUA-EDC/NHS). The developed phage sensor was characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), scanning electron microscopy (SEM), electrochemical analysis using cyclic voltammetry (CV), and square wave voltammetry (SWV). Under optimal conditions, the affinity peptide-displayed phage sensor showed a good binding affinity (Kd = 0.13 ± 0.56 µM) and limit of detection (0.39 µM) for caspase-3 detection. Furthermore, developed phage sensor could be monitored the response of apoptotic HeLa cells by detecting caspase-3 activity. This work should stimulate the development of efficient alternative caspase-3 detection methods for the diagnosis and prognosis of apoptosis-related diseases.

Facile synthesis of carbon dots from Tagetes erecta as a precursor for determination of chlorpyrifos via fluorescence turn-off and quinalphos via fluorescence turn-on mechanisms.

Convenient one-pot synthetic route for the fabrication of carbon dots (CDs) from Tagetes erecta flower (TEF), named as "TEF-CDs', through solvo(hydro)-thermal carbonization of the TEF was developed. The TEF-CDs revealed high selectivity towards chlorpyrifos and quinalphos, acting as a fluorescent probe. The CDs synthesized from T. erecta flower showed a strong blue color at 495 nm when excited at 420 nm, along with the exhibition of a strong quantum yield of 63.7%. The synthesized CDs revealed their richness in the surface-active organic group that synthesized CDs from T. erecta flower are mainly comprised of C, O, and N, which were crystalline in structure that was revealed by TEM image and XRD spectra. Furthermore, when the probe was exposed to different pH conditions, no major noticeable changes were recorded. Moreover, when the probe was exposed to chlorpyrifos and quinalphos, we have noticed that fluorescence spectra was turned off when the probe was exposed to chlorpyrifos and "turned on" after the exposure quinalphos. Moreover, fluorescence spectral changes showed a good linearity with chlorpyrifos and quinalphos concentrations in the range of 0.05-100.0 µM for chlorpyrifos and 0.01-50.0 µM for quinalphos. The limit of detection are 2.1 ng mL-1 and 1.7 ng mL-1 for chlorpyrifos and quinalphos, respectively. Finally, the TEF-CDs-based fluorescent nanoprobe was successfully applied to estimate chlorpyrifos and quinalphos with an effective accuracy in rice and fruit samples with rapid detection time.

Photo-induced reactions for disassembling of coloaded photosensitizer and drug molecules from upconversion-mesoporous silica nanoparticles: An effective synergistic cancer therapy.

Photodynamic therapy is an emerging noninvasive cancer treatment approach, which requires a photosensitizer (PS), light, and molecular oxygen. In this study, we have successfully fabricated a dual nature (pH- and reactive-oxygen-species-responsive) upconversion nanoparticles (UCNPs) to utilize coloaded doxorubicin (DOX) and chlorin e6 (Ce6) with high antitumor efficacy. The model anticancer drug (DOX) and PS (Ce6) were conjugated in a ratio of 1:1 (w:w), and then loaded on the surface of UCNPs@mesoporous silica (mSiO2) (85.63 ± 9.87 nm). Cellular uptake could be achieved by either increased permeability or ionic effect of UCNPs@mSiO2, where Ce6 controlled the DOX release under a near-infrared (NIR) laser irradiation at 980 nm. A cytotoxicity analysis revealed that the dual-responsive UCNPs@mSiO2 could successfully deliver DOX and Ce6 at the tumor site, causing cell death with a high efficiency. This study shows that the modified UCNPs@mSiO2 is a promising system to realize NIR-light-triggered PS and drug delivery approach to improve synergistic therapies in vitro and in vivo, in the future.

One-pot synthesis of carbon dots with intrinsic folic acid for synergistic imaging-guided photothermal therapy of prostate cancer cells.

Photothermal therapy (PTT) is performed using near-infrared-responsive agents, which is proven to be an effective therapeutic strategy against cancer with several advantages including minimal invasion, high effectiveness, and easy implementation. Herein, we report a facile and novel one-pot synthetic approach for the fabrication of polydopamine-folate carbon dots (PFCDs) as theranostic nanocarriers for the image-guided PTT targeting of prostate cancer (PCa) cells that express a prostate-specific membrane antigen (PSMA) (folate hydrolase 1). The as-fabricated PFCDs exhibited several advantages such as easy preparation, high biocompatibility, low toxicity, good water-solubility, and excellent photothermal effect with robust blue fluorescence emission. The PSMA-directed imaging of PCa using PFCDs showed remarkable fluorescence enhancement in LNCap cells as compared to the case of other cells that did not express PSMA. PFCDs exhibited a photothermal effect in the PCa cells when irradiated with an 808 nm laser, which possibly resulted in the complete elimination of the tumor. Thus, these features make PFCDs a promising candidate for PTT. Moreover, PFCD-based PTT provides an effective biomedical platform for cancer therapy.

A facile hydrothermal synthesis of highly luminescent NaYF4:Yb3+/Er3+ upconversion nanoparticles and their biomonitoring capability.

Using a facile hydrothermal procedure, hydrophilic NaYF4: Yb3+/Er3+ nanoparticles (NPs) have been prepared as lanthanide-doped upconversion (UC) materials exhibiting different morphologies, crystal phases and luminescence intensity. The upconversion nanoparticles (UCNP) were characterized by means of electron microscopy and spectroscopy, X-ray diffraction (XRD) and photoluminescence analysis. The molar concentration of reactants and volumes of NaF affect the shapes and uniformity of the synthesized NPs. These parameters also have influence on crystal phase and luminescence intensity of the NPs. Adjusting hydrothermal reaction time and dopant concentration also enable the synthesis of NPs with strong UC luminescence. The as-prepared UCNP showed cellular nontoxicity to HeLa cells, and thus they are capable as promising agents for biological imaging.

Morphological evolution of upconversion nanoparticles and their biomedical signal generation.

Advancements in the fabrication of upconversion nanoparticles (UCNPs) for synthetic control can enable a broad range of applications in biomedical systems. Herein, we experimentally verified the role of the hydrothermal reaction (HR) time in the synthesis of NaYF4:20%Yb3+/3%Er3+ UCNPs on their morphological evolution and phase transformation at different temperatures. Characterizations of the as-prepared UCNPs were conducted using X-ray diffraction (XRD), electron microscopy and spectroscopy, and thermogravimetric and upconversion (UC) luminescence analysis. We demonstrated that determining the optimal HR time, also referred to here as the threshold time, can produce particles with good homogeneity, hexagonal phase, and UC luminescence efficiency. Subsequently, the polymer coated UCNPs maintained their original particle size distribution and luminescence properties, and showed improved dispersibility in a variety of solvents, cellular nontoxicity, in vitro bioimaging, and biocompatibility as compared to the bare UCNP. Besides this, polyacrylic acid conjugated UCNPs (UCNP@PAA) also revealed the strong anticancer effect by conjugating with doxorubicin (DOX) as compared to the free DOX. Based on these findings, we suggest that these particles will be useful in drug-delivery systems and as in vivo bioimaging agents synchronously.