Plant-Derived Anti-Human Epidermal Growth Factor Receptor 2 Antibody Suppresses Trastuzumab-Resistant Breast Cancer with Enhanced Nanoscale Binding.

Traditional monoclonal antibodies such as Trastuzumab encounter limitations when treating Human Epidermal Growth Factor Receptor 2 (HER2)-positive breast cancer, particularly in cases that develop resistance. This study introduces plant-derived anti-HER2 variable fragments of camelid heavy chain domain (VHH) fragment crystallizable region (Fc) KEDL(K) antibody as a potent alternative for overcoming these limitations. A variety of biophysical techniques, in vitro assays, and in vivo experiments uncover the antibody's nanoscale binding dynamics with transmembrane HER2 on living cells. Single-molecule force spectroscopy reveals the rapid formation of two robust bonds, exhibiting approximately 50 pN force resistance and bond lifetimes in the second range. The antibody demonstrates a specific affinity for HER2-positive breast cancer cells, including those that are Trastuzumab-resistant. Moreover, in immune-deficient mice, the plant-derived anti-HER2 VHH-FcK antibody exhibits superior antitumor activity, especially against tumors that are resistant to Trastuzumab. These findings underscore the plant-derived antibody's potential as an impactful immunotherapeutic strategy for treating Trastuzumab-resistant HER2-positive breast cancer.

Exosome-Coated Prussian Blue Nanoparticles for Specific Targeting and Treatment of Glioblastoma.

Glioblastoma is one of the most aggressive and invasive types of brain cancer with a 5-year survival rate of 6.8%. With limited options, patients often have poor quality of life and are moved to palliative care after diagnosis. As a result, there is an extreme need for a novel theranostic method that allows for early diagnosis and noninvasive treatment as current peptide-based delivery standards may have off-target effects. Prussian Blue nanoparticles (PBNPs) have recently been investigated as photoacoustic imaging (PAI) and photothermal ablation agents. However, due to their inability to cross the blood-brain barrier (BBB), their use in glioblastoma treatment is limited. By utilizing a hybrid, biomimetic nanoparticle composed of a PBNP interior and a U-87 cancer cell-derived exosome coating (Exo:PB), we show tumor-specific targeting within the brain and selective thermal therapy potential due to the strong photoconversion abilities. Particle characterization was carried out and showed a complete coating around the PBNPs that contains exosome markers. In vitro cellular uptake patterns are similar to native U-87 exosomes and when exposed to an 808 nm laser, show localized cell death within the specified region. After intravenous injection of Exo:PB into subcutaneously implanted glioblastoma mice, they have shown effective targeting and eradication of tumor volume compared to PEG-coated PBNPs (PEG:PB). Through systemic administration of Exo:PB particles into orthotopic glioblastoma-bearing mice, the PBNP signal was detected in the brain tumor region through PAI. It was seen that Exo:PB had preferential tumor accumulation with less off-targeting compared to the RGD:PB control. Ex vivo analysis validated specific targeting with a direct overlay of Exo:PB with the tumor by both H&E staining and Ki67 labeling. Overall, we have developed a novel biomimetic material that can naturally cross the BBB and act as a theranostic agent for systemic targeting of glioblastoma tissue and photothermal therapeutic effect.

Thermally Induced Morphological and Structural Transformations on Eu2+/Eu3+-Coactivated Calcium Silicate Nanophosphors.

This study presents an approach for synthesizing Eu2+/Eu3+-coactivated Ca2SiO4 nanophosphors, by adjusting the ratio of both activators within a singular host material. Utilizing a hydrothermal method complemented by a postreduction sintering process, we fabricated a series of phosphors characterized by uniform 30-50 nm spherical nanoparticles. These engineered phosphors manifest multichannel luminescence properties and exhibit simultaneous blue and red emission from Eu2+ and Eu3+, respectively. Meticulous control of the 5% H2-95% N2 reduction temperature allowed for precise tuning of the Eu2+ and Eu3+ ions within the host lattice, which enabled the strategic adjustment of their luminescent outputs. Utilizing X-ray photoelectron spectroscopy (XPS), we could discern subtle alterations in the europium oxidation state. By using a transmission electron microscope (TEM) and an X-ray diffractometer (XRD), we found that the subsequent changes by reductive sintering to particle size, morphology, and mixed crystal structures influenced the materials' luminescent characteristics. Our findings herald a significant advancement in solid-state lighting, with the potential for the use of Eu2+/Eu3+-coactivated calcium silicate nanophosphors to develop white light emission technologies endowed with enhanced color rendering and luminous efficacy.

Carbon Coated Iron-Cobalt Nanoparticles for Magnetic Particle Imaging.

Magnetic particle imaging (MPI) is an emerging imaging modality that provides direct and quantitative mapping of iron oxide tracers. To achieve high sensitivity and good spatial resolution images, a magnetic nanoparticle with a higher contrast intensity needs to be developed. Currently, a majority of MPIs being developed for potential clinical application are composed of iron oxide nanoparticles with a spherical shape. In this project, we intend to report development of high-performance carbon (C) coated iron-cobalt (FeCo) nanoparticles (FeCo/C) and investigate their feasibility as a MPI agent. We have synthesized FeCo/C through a facile and simple method at mild temperature that is safe, easy, and up-scalable. We studied the structural and functional relationships and biocompatibility of this MPI agent in vitro. However, to enhance the aqueous solubility and biocompatibility, the surface of FeCo/C was modified with polyethylene glycol (PEG). We found that variation in the ratio of Fe and Co plays a vital role in their physical properties and functionality. In vitro imaging confirms that the Fe3Co1/C nanoparticle has highly competitive MPI intensity compared to VivoTrax, a commercially available MPI agent. Confocal laser scanning microscopy imaging with Rhodamine B labeled FeCo/C displays cellular internalization by the A375 cancer cells. The in vitro toxicity analysis concludes that there is no significant toxicity of FeCo/C nanoparticles. Therefore, the newly developed MPI agent holds strong promise for biomedical imaging and could be further validated in vivo in small animals.

Europium-Doped Calcium Silicate Nanoparticles as High-Quantum-Yield Red-Emitting Phosphors.

Europium ion-activated calcium silicate phosphors (Ca2SiO4:Eu3+) with sharp red-light emission were fabricated via the hydrothermal method. The size of Ca2SiO4:Eu3+ phosphors was controlled between 20 and 200 nm by precursor silicate particle sizes. Systematic studies to determine morphology, crystal phase, and photoluminescence (PL) were carried out for all the phosphors, and their optical efficiencies were compared. We found that the luminescence intensity and emission wavelength of Ca2SiO4:Eu3+ phosphors depend on their particle sizes. Particularly, the Ca2SiO4:Eu3+ synthesized with 20 nm silica seed contains the most intense red emission, high color purity, and high PL quantum yield. For the 20 nm-sized Ca2SiO4:Eu3+ phosphor, PL quantum yields are measured to be above 87.95% and high color purity of 99.8%. The unusually high intensity of 5D0 → 7F4 emission (712 nm) is explained by structural distortion arising from silicate particle size reductions. We show that the obtained phosphor is a suitable candidate for solid-state lighting as a red component through CIE chromaticity coordinate and color purity measurements. Furthermore, the Ca2SiO4:Eu3+ particles are examined for their validity as promising bio-imaging probes through cell labeling and imaging experiments and biodegradability studies.

Hovenia Monofloral Honey can Attenuate Enterococcus faecalis Mediated Biofilm Formation and Inflammation.

We evaluated the anti-biofilm formation and anti-inflammatory activity of Hovenia monofloral honey (HMH) against Enterococcus faecalis. Co-culture of HMH with E. faecalis attenuated the biofilm formation of E. faecalis on a polystyrene surface. In addition, HMH effectively eradicated the established E. faecalis biofilm. HMH significantly attenuated E. faecalis growth but did not affect the production of extracellular polymeric substances on E. faecalis, indicating that reduction of E. faecalis biofilm is a result of HMH-mediated killing of E. faecalis. Furthermore, we found that HMH can effectively attenuate E. faecalis-induced expression of a proinflammatory interleukin-8 (IL-8) in HT-29 cells. Interestingly, treatment of HMH significantly attenuated the E. faecalis-mediated expression of Toll-like receptor-2 (TLR-2) and its adaptor molecules, myeloid differentiation primary response 88 (MyD88), in HT-29 cells. In addition, E. faecalis-induced mitogen-activated protein kinases (MAPKs) phosphorylation was significantly attenuated by HMH administration. Furthermore, HMH-mediated anti-inflammatory efficacy (0.2 mg/mL of HMHs) had an equal extent of inhibitory efficacy as 5 µM of MyD88 inhibitor to attenuate E. faecalis-mediated IL-8 expression in HT-29 cells. These results suggest that HMH could effectively inhibit E. faecalis-mediated gastrointestinal inflammation through regulating the TLR-2/MyD88/MAPKs signaling pathways. Collectively, our data suggest that HMH could be developed as a potential natural agent to control E. faecalis-mediated biofilm formation and inflammation.

An investigation on Eu3+ doped CaLa2ZnO5 novel red emitting phosphors for white light-emitting diodes.

A novel phosphor namely CaLa2ZnO5 doped with Eu3+ ions were prepared by conventional solid state reaction method. We have studied and optimized various constraints like sintering temperature, sintering time and dopant concentration. XRD, SEM profiles have been studied to explore its structural properties. Luminescence properties of these phosphors have been characterized by means of their photoluminescence (PL) spectra. We have noticed that the emission intensity of CaLa2ZnO5:Eu3+ phosphors strongly depend on its sintering temperature and Eu3+ concentration. Moreover, their PL spectra reveals that CaLa2ZnO5:Eu3+ phosphors exhibits a strong luminescence of 5D(0)_7F(2) transition at 627 nm under the excitation of 468 nm, which correspond to the popular emission line from a GaN based blue light-emitting diode (LED) chip. The obtained results of the prepared Eu3+ doped phosphors are very much encouraging and they are potentially useful in the development of new solid-state lightning devices.

Induction of apoptosis and inhibition of telomerase activity by trichostatin A, a histone deacetylase inhibitor, in human leukemic U937 cells.

The objective of the present study was to investigate the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on the cell growth and apoptosis and its effect on the telomerase activity in human leukemic cell line U937. Exposure of U937 cells to TSA resulted in growth inhibition and induction of apoptosis in a dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy, agarose gel electrophoresis and flow cytometry analysis. The increase in apoptosis was associated with the up-regulation in proapoptotic Bax expression and down-regulation of antiapoptotic Bcl-2 and Bcl-X(L). TSA treatment inhibited the levels of cIAP family members and induced the proteolytic activation of caspase-3, which was associated with concomitant degradation of poly(ADP-ribose)-polymerase and beta-catenin protein. TSA treatment markedly inhibited the activity of telomerase in a dose-dependent fashion. Additionally, the expression of human telomerase reverse transcriptase (hTERT), a main determinant of the telomerase enzymatic activity, was progressively down-regulated by TSA treatment. We therefore conclude that TSA demonstrated antiproliferative and apoptosis-inducing effects on U937 cells in vitro, and that changes in Bcl-2 family protein levels as well as telomerase activity may play an important role in its mechanism of action.

Beta-lapachone, a quinone isolated from Tabebuia avellanedae, induces apoptosis in HepG2 hepatoma cell line through induction of Bax and activation of caspase.

The DNA topoisomerase inhibitor beta-lapachone is a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America. It has been reported to possess a wide range of pharmacological properties, and is a promising cancer chemopreventive agent. In this study, the effects of beta-lapachone on the growth of the human hepatoma cell line HepG2 were investigated. The results showed that beta-lapachone inhibits the viability of HepG2 by inducing apoptosis, as evidenced by the formation of apoptotic bodies and DNA fragmentation. Reverse transcription-polymerase chain reaction and immunoblotting results indicated that treatments of cells with beta-lapachone resulted in down-regulation of anti-apoptotic Bcl-2 and Bcl-X(L) and up-regulation of pro-apoptotic Bax expression. beta-Lapachone-induced apoptosis was associated with a proteolytic activation of caspase-3 and -9 and degradation of poly(ADP-ribose) polymerase protein. However, beta-lapachone treatment did not affect the inhibitor of apoptosis proteins family and the Fas/FasL system. Taken together, our study indicated that beta-lapachone may have potential as a chemopreventive agent for liver cancer.

Growth inhibition of A549 human lung carcinoma cells by beta-lapachone through induction of apoptosis and inhibition of telomerase activity.

The objective of the present study was to investigate the effect of beta-lapachone, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae), on the cell growth and apoptosis in human lung carcinoma cell line A549. Exposure of A549 cells to beta-lapachone resulted in growth inhibition and induction of apoptosis in a time- and dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometry analysis. This increase in apoptosis was associated with a decrease in Bcl-2 and expression, an increase of Bax, and an activation of caspase-3 and caspase-9. beta-lapachone treatment markedly inhibited the activity of telomerase in a dose-dependent fashion. Additionally, the levels of human telomerase RNA (hTR) and c-myc expression were progressively down-regulated by beta-lapachone treatment. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of beta-lapachone.

Escherichia coli 6-pyruvoyltetrahydropterin synthase ortholog encoded by ygcM has a new catalytic activity for conversion of sepiapterin to 7,8-dihydropterin.

The putative gene (ygcM) of Escherichia coli was verified in vitro to encode the ortholog of 6-pyruvoyltetrahydropterin synthase (PTPS). Unexpectedly, the enzyme was found to convert sepiapterin to 7,8-dihydropterin without any cofactors. The enzymatic product 7,8-dihydropterin was identified by HPLC and mass spectrometry analyses, suggesting a novel activity of the enzyme to cleave the C6 side chain of sepiapterin. The optimal activity occurred at pH 6.5-7.0. The reaction rate increased up to 3.2-fold at 60-80 degrees C, reflecting the thermal stability of the enzyme. The reaction required no metal ion and was activated slightly by low concentrations (1-5 mM) of EDTA. The apparent K(m) value for sepiapterin was determined as 0.92 mM and the V(max) value was 151.3 nmol/min/mg. The new catalytic function of E. coli PTPS does not imply any physiological role, because sepiapterin is not an endogenous substrate of the organism. The same activity, however, was also detected in a PTPS ortholog of Synechocystis sp. PCC 6803 but not significant in Drosophila and human enzymes, suggesting that the activity may be prevalent in bacterial PTPS orthologs.

Production of sepiapterin in Escherichia coli by coexpression of cyanobacterial GTP cyclohydrolase I and human 6-pyruvoyltetrahydropterin synthase.

Synechocystis sp. strain PCC 6803 GTP cyclohydrolase I and human 6-pyruvoyltetrahydropterin synthase were coexpressed in Escherichia coli. The E. coli transformant produced sepiapterin, which was identified by high-performance liquid chromatography and enzymatically converted to dihydrobiopterin by sepiapterin reductase. Aldose reductase, another indispensable enzyme for sepiapterin production, may be endogenous in E. coli.

Functional investigation of a gene encoding pteridine glycosyltransferase for cyanopterin synthesis in Synechocystis sp. PCC 6803.

A gene (slr1166) putatively encoding pteridine glycosyltransferase was disrupted with a kanamycin resistance cassette in Synechocystis sp. PCC 6803, which produces cyanopterin. The deduced polypeptide from slr1166 consisted of 354 amino acid residues sharing 45% sequence identity with UDP-glucose:tetrahydrobiopterin alpha-glucosyltransferase (BGluT) isolated previously from Synechococcus sp. PCC 7942. The knockout mutant was unable to produce cyanopterin but only 6-hydroxymethylpterin-beta-galactoside, verifying that slr1166 encodes a pteridine glycosyltransferase, which is responsible for transfer of the second sugar glucuronic acid in cyanopterin synthesis. The mutant was affected in its intracellular pteridine content and growth rate, which were 74% and 80%, respectively, of wild type, demonstrating that the second sugar residue is still required for quantitative maintenance of cyanopterin. This supports the previous suggestion that glycosylation may contribute to high cellular concentration of pteridine compounds.