Omic Evaluation of Nanomaterial-Based Photodynamic Therapy of Cancer.

Photodynamic therapy (PDT), a noninvasive cancer treatment, relies on three components: light source, oxygen, and photosensitizer (PS). When PS is excited by a specific wavelength of light in the presence of oxygen, it leads to the generation of reactive oxygen species (ROS), which results in targeted destruction of cancer cells. The success of PDT mainly depends on the properties of the chosen PS, emphasizing selectivity, high absorbance, drug conjugation, controlled biodistribution, and low toxicity. Nanomaterials not only play an important role in photochemical activity by maximizing the absorption of photons from the light source but can also adjust the pharmacokinetics and tumor selectivity of photoactive molecules. Therefore, they can be used as a PS on their own and conjugated with other PS molecules. When combined with selectivity, high targeting capacity, and finally, light of the appropriate wavelength, the scenario results in localized ROS formation and cell death. However, the signaling pathways of PDT-induced cell death may differ depending on the cell type or nanomaterial properties. For this reason, omics analyses are needed to clarify the mechanisms underlying photodynamic reactions. Proteomics, crucial in molecular sciences, sheds light on cancer mechanisms, identifying biomarkers and therapeutic targets. Examining nanoparticle-based PDT in cancer cell lines in vitro, this chapter aims to molecularly evaluate efficacy, utilizing proteomic analysis to understand the underlying mechanisms.

Transcriptomic evaluation of metals detected in placenta.

This research aims to assess the concentration of metals in human and canine placentas from the same geographic area and to investigate how these metal levels influence gene expression within the placenta. Placentas of 25 dogs and 60 women who had recently given birth residing in Ankara, Turkey were collected and subjected to metal analysis using ICP-OES. Placentas with detectable metal levels underwent further examination including Next Generation Sequencing, transcriptional analysis, single nucleotide polymorphism investigation, and extensive scrutiny across various groups. For women, placentas with concurrent detection of aluminum (Al), lead (Pb), and cadmium (Cd) underwent transcriptomic analysis based on metal analysis results. However, the metal load in dog placentas was insufficient for comparison. Paired-end sequencing with 100-base pair read lengths was conducted using the DNBseq platform. Sequencing quality control was evaluated using FastQC, fastq screen, and MultiQC. RNA-sequencing data is publicly available via PRJNA936158. Comparative analyses were performed between samples with detected metals and "golden" samples devoid of these metals, revealing significant gene lists and read counts. Normalization of read counts was based on estimated size factors. Principal Component Analysis (PCA) was applied to all genes using rlog-transformed count data. Results indicate that metal exposure significantly influences placental gene expression, impacting various biological processes and pathways, notably those related to protein synthesis, immune responses, and cellular structure. Upregulation of immune-related pathways and alterations in protein synthesis machinery suggest potential defense mechanisms against metal toxicity. Nonetheless, these changes may adversely affect placental function and fetal health, emphasizing the importance of monitoring and mitigating environmental exposure to metals during pregnancy.

Effects of DDT and DDE on placental cholinergic receptors.

The placental cholinergic system; known as an important factor in intracellular metabolic activities, regulation of placental vascular tone, placental development, and neurotransmission; can be affected by persistent organic pesticides, particularly organochlorine pesticides(OCPs), which can influence various epigenetic regulations and molecular pathways. Although OCPs are legally prohibited, trace amounts of the persistent dichlorodiphenyltrichloroethane(DDT) are still found in the environment, making prenatal exposure inevitable. In this study, the effects of 2,4'-DDT and 4,4'-DDT; and its breakdown product 4,4'-DDE in the environment on placental cholinergic system were evaluated with regards to cholinergic genes. 40 human placentas were screened, where 42,50% (17 samples) were found to be positive for the tested compounds. Average concentrations were 10.44 µg/kg; 15.07 µg/kg and 189,42 µg/kg for 4,4'-DDE; 2,4'-DDT and 4,4'-DDT respectively. RNA-Seq results revealed 2396 differentially expressed genes in positive samples; while an increase in CHRM1,CHRNA1,CHRNG and CHRNA2 genes at 1.28, 1.49, 1.59 and 0.4 fold change were found(p<0028). The increase for CHRM1 was also confirmed in tissue samples with immunohistochemistry. In vitro assays using HTR8/SVneo cells; revealed an increase in mRNA expression of CHRM1, CHRM3 and CHRN1 in DDT and DDE treated groups; which was also confirmed through western blot assays. An increase in the expression of CHRM1,CHRNA1, CHRNG(p<0001) and CHRNA2(p<0,05) were found from the OCPs exposed and non exposed groups.The present study reveals that intrauterine exposure to DDT affects the placental cholinergic system mainly through increased expression of muscarinic receptors. This increase in receptor expression is expected to enhance the sensitivity of the placental cholinergic system to acetylcholine.

Silk fibroin nanoparticles and ß-tricalcium phosphate loaded tissue engineered gelatin bone scaffolds: A Nature-based, low-cost solution.

Tissue engineering has recently attracted attention as an alternative to traditional treatment methods for tissue and organ damage. Since bone is one of the most important vital parts of the body, the treatment of bone damage is important. Silk fibroin is a natural polymer with properties such as biocompatibility and biodegradability, which attracts attention with its controlled release, especially in drug delivery systems. In this study, gelatin-based scaffolds loaded with silk fibroin nanoparticles and ß -tricalcium phosphate (ß -TCP) were developed to be used as a potential drug delivery system in bone tissue engineering. The chosen nanoparticle formulation has a 294 nm average diameter with a 0.380 polidispersity index (PDI). In vitro characterization of scaffolds was performed by mechanical, morphological characterization, swelling capacity, Differential Scanning Calorimetry (DSC), Fourier-Transform Infrared Spectroscopy (FT-IR) measurements, and biocompatibility was evaluated by cell culture studies. Swelling index, tensile strength, elongation at break, and Young modulus of the ß -TCP and silk nanoparticles loaded scaffold were found as 456%, 1.476 MPa, 6.75%, and 24 MPa, respectively. In vitro cell culture studies have shown that scaffolds prepared in the present study can accelerate osteoblast differentiation and increase the healing rate of bone tissues. In addition, they have the potential to be used as a drug delivery system in bone tissue engineering that needs to be evaluated with further studies.

Apis mellifera anatoliaca Venom Exerted Anti-Inflammatory Activity on LPS-Stimulated Mammalian Macrophages by Reducing the Production of the Inflammatory Cytokines.

Extraction and characterization of natural products provide the opportunity to expand our arsenal of drug candidates against a wide range of diseases including cancer and inflammatory disorders. Previous studies have shown bee venom to have immense potential as an anti-inflammatory drug candidate. In this study, we focused on the venom of Apis mellifera anatoliaca and characterized its content by HPLC. An in vitro inflammation model based on lipopolysaccharide (LPS)-stimulated mammalian macrophages was utilized to examine the venom's anti-inflammatory potential. Additionally, its antiproliferative activity was evaluated in vitro against a human glioblastoma cell line. Based on the TNF, IL6, GMCSF, and IL12p40 pro-inflammatory cytokine production level in LPS-induced macrophages, venom-treated groups showed substantial decrease in the inflammatory action compared to untreated LPS-stimulated macrophages. When the cells were analyzed for viability, the venom did not have any cytotoxic effect on the macrophages at the concentration ranges that were utilized. Moreover, IC50 value of the venom was above 60 µg/mL on glioblastoma cancer cell line. These results suggest that the Apis mellifera anatoliaca venom does not have anticancer drug candidate potential, whereas it can efficiently be used against inflammatory and autoimmune disorders. To our knowledge, this is the first study to specifically examine the effect of anti-inflammatory activity of Apis mellifera anatoliaca venom on macrophages.

Live in same region, respond differently: Canine and human response to pollutants in placental accumulation.

Polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), and polycyclic aromatic hydrocarbons (PAHs) are endocrine-disrupting chemicals (EDCs). Their presence in the environment is particularly concerning in cases of fetal exposure, which is the most vulnerable period of life for both human and animals who share the same environment. Placenta, as a sample collected using noninvasive methods to screen EDCs, is a good indicator for potential fetal exposure. Although recent studies indicate that companion animal may correspond to human exposure, species-specific anatomo-morphological and metabolic differences are controversial. In this study, placenta samples of 60 women and 25 dogs living and giving birth within the same region were evaluated for the presence of PCB, OCP, PBDE, and PAH residues; where, socio-demographic factors were also assessed to identify the possible sources. Gas chromatography-mass spectrometry method was validated for the matrix, and among 45 screened and targeted pollutants, only 18 were found in human placentas. While the most frequently detected pollutants were DDTs, followed by PAHs and PCBs in decreasing order, the pollutants with the highest concentrations were PAHs, followed by PCBs and DDTs. Only five of the target contaminants were detected in the dog placentas. These results indicate that; as dogs have different bioaccumulation capacities and higher excretion rates than humans, the life-long effects of exposure to endocrine compound and possible consequences related to adverse health outcomes are expected to vary and concentrations cannot be directly correlated.

Diethylhexyl Phthalate and Bisphenol A Promote Vincristine and Tamoxifen Resistance in Vitro.

Environmental estrogen active compounds are strong determinants of estrogen receptor (ER)-positive breast cancers, and increased evidence indicates their contribution to chemotherapy resistance. In the current study, the efficacy of vincristine and tamoxifen, with the presence of diethylhexyl phthalate (DEHP) and bisphenol A (BPA) and the possible involvement of estrogen and estrogen receptor-related mechanisms, was evaluated in an ER+ mammary tumor cancer cell line, MCF-7. Chemotherapeutics tamoxifen as an estrogen receptor modulator and vincristine as an antimitotic compound were selected for evaluation against the presence of common endocrine disrupters. BPA and DEHP preincubation at their proliferative concentrations for 4 h was found to decrease the cytotoxicity of vincristine. mRNA and protein expression of ESR1 and ESR 2 were decreased by vincristine, while this decrease was reversed by DEHP and BPA. Both BPA and DEHP were able to interfere with the cytotoxic activity of vincristine against MCF-7 cells through ESR1 and ESR2. This study provides in vitro toxicological evidence for vincristine resistance and its relation to estrogen active environmental pollutants in ER+ breast cancer cells.

Effects of electrospun fiber curcumin on bisphenol A exposed Caco-2 cells.

Curcumin; the major polyphenolic compound, isolated from Curcuma longa L.; loaded polyvinylpyrrolidone K90 fibers were prepared using electrospinning method. Effectiveness was tested on human colorectal adenocarcinoma cells with the presence of the endocrine disrupter Bisphenol A. Curcumin-loaded fibers were shown to have good physicochemical properties where excellent morphology of the electrospin fibers were formed. With the presence of 8 nM Bisphenol A, 17.37 mM fibers were found to inhibit proliferation in the cells in a dose-dependent manner. Fibers induced a significant increase in malondialdehyde by Thiobarbituric Acid Reactive Substances Assay compared to the control and this effect was supported by the presence of Bisphenol A. Western blot results indicate Super Oxide Dismutase-1 levels were increased by fiber, while Bisphenol A coincubated group resulted in a decrease. Fibers increased the expression of Estrogen Receptor 2, while Estrogen Receptor 1 expression did not change. Estrogen Receptor 2 expression was increased by coincubation with Bisphenol A; indicating a possible role of Estrogen Receptor 2 in the protective effects of fiber. This study presents that fiber had enhanced bioavailability and solubility with increased anticancer effect in human colon adenocarcinoma cells in presence of Bisphenol A; where involved mechanisms are antioxidant system and estrogen receptor expression.