Volume 49, Issue 5

Diabetes mellitus (DM) represents a heterogeneous and multifactorial metabolic disorder, leading to diverse economic challenges and health complications, significantly affecting individuals’ lives. Type 2 diabetes (T2DM) stands as the predominant form of diabetes, with its onset influenced by a range of genetic and environmental factors. Currently, conventional diabetic medications are employed in clinical practice, but they may entail certain side effects. Fortunately, diverse natural polysaccharides extracted from plants exhibit antidiabetic properties, which have been demonstrated to effectively mitigate hyperglycemia, reverse insulin resistance, and forestall complications associated with diabetes. Consequently, exploring the utilization of these polysaccharides as adjuncts to conventional therapies has emerged as a novel research direction in recent years. Thus, this review centers on the extraction, purification, and characterization of plant polysaccharides, providing an overview of the advancements in the antidiabetic effects and mechanisms of natural polysaccharides. This perspective offers fresh insights to explore potential research domains and enhances understanding for the development of diabetic drugs derived from polysaccharides.

Objectives It has been recently reported that immune activity of some animal peptides/proteins has been found in some plants. In this study, the presence of immune reactivity of alarin, asprosin, paraoxonase, elabela, glucagon, and nesfatin-1 like molecules was investigated in two plant species related to Kastamonu Taşköprü garlic (an endemic cultivar of the Allium sativum L.) and Tunceli garlic ( Allium tuncelianum ). Methods Ten Kastamonu and 10 Tunceli garlic were embedded in paraffin blocks and 4–5 μm transversal and longitudinal sections were taken and stained by immunohistochemical method. In addition, 500 mg of wet tissue samples were homogenized from these garlic and supernatants were obtained. The amounts of alarin, asprosin, elabela, glucagon and nesfatin-1 were studied by enzyme linked immunosorbent assay (ELISA) method and paraoxonase was studied by spectrophotometric method. Results Studied molecules show immune reactivity in the cytoplasm of the bulb storage cells of the plants parenchyma. The ELISA supernatant results also confirmed the presence of these animal molecules in the plants. Conclusions Although we cannot reveal the purpose of the presence of these animal molecules detected in plants with these existing data, we predict that these data will be a source of inspiration in the field of botany, that studies will shift in this direction and that they might contribute to the revision of the evolutionary processes of all living things, especially an ancestor of garlics.

Objectives The primary aim of this study is to immobilize glucose oxidase (GOX) as a model enzyme using a simple and efficient method without the use of any toxic chemicals. This will enable the multiple reusability of GOX, thereby reducing the process cost. Furthermore, the objective is to determine the kinetic, optimum pH, and temperature parameters in comparison to soluble GOX. Methods For immobilization, GOX at a concentration of 5 mg/mL was mixed with an equal volume of hypol prepolymer in a beaker. Immobilization was achieved by the formation of amide bonds between –NH 2 groups on the enzyme surface and the prepolymer. The activities of polyurethane foam (PUF)-immobilized and soluble GOX were monitored using horse radish peroxidase enzyme as a second enzyme. 2,2′-Azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical ABTS + generated as a result of the enzymatic reactions was detected at 420 nm. Results The kinetic parameters for soluble GOX were Km=9.39 ± 1.13 mM and V max =9.14 ± 0.45 A.U sec −1  × 10 −4 , while for PUF-immobilized GOX Km=10.04 ± 1.30 mM and V max =9.09 ± 0.49 A.U sec −1  × 10 −4 . PUF-immobilized GOX displayed high performance during reuse experiments. PUF-immobilization led to the activity at higher temperatures and over a wide pH range compared to its soluble counterpart. Conclusions GOX was successfully immobilized for the first time using PUF. This technique can be used in the healthcare and food sectors as well as in biosensor applications. In the future, when successfully applied to many enzymes of industrial value, it could offer significant advantages.

Objectives Sulforaphane is well-known for its anti-cancer properties particularly against breast, skin and prostate cancers. High sensitivity of sulforaphane to oxygen, heat, and alkaline conditions, as well as its poor oral bioavailability and water instability limit its use in medicine. In this study, we aim to overcome the prementioned limitations by encapsulating sulforaphane within a lipid-based drug delivery system, known as emulsome, and investigate the anti-cancer features of the attained formulation. Methods The stability and dispersity of the formulation were assessed sequentially by zeta sizer, scanning electron microscopy and confocal laser scanning microscopy. Cell culture studies were performed to evaluate the anticancer activity of the formulation. Results Sulforaphane-loaded emulsomes with an average particle size of 246.0±14.1 nm, an average zeta potential of −23.5±2.4 mV and a polydispersity index of around 0.38 were produced. Encapsulations up to 0.036 mg/mL sulforaphane concentration was achieved. When MCF7 breast cancer cells were treated with sulforaphane-loaded emulsomes, a significant decrease was observed in proliferation of the cells along 72 h. In control group, emulsomes were found safe as tested at same concentrations on MCF-10a healthy cells. Applied as dissolved in DMSO, free sulforaphane with an IC 50 value of 1.2 µM was more effective against MCF7 cells than sulforaphane-loaded emulsome formulation having a IC 50 value 21.1 µM. Conclusions Sulforaphane-loaded emulsomes were obtained as stable, moderately disperse suspensions. Delivery of the bioactive compound into the cells were achieved. Yet, its biological activity remained behind its free form.

Objectives Cholangiocarcinoma (CCA) is a highly heterogeneous biliary malignant tumor. Studies have demonstrated that JAK/STAT signaling is activated in many types of cancer. In addition, JAK/STAT is activated downstream of AXL, and the AXL receptor is activated by its ligand, GAS6. In this study, we investigated the anticarcinogenic effect of silibinin and its relationship with the GAS6/AXL ve JAK/STAT pathway in the human EGI-1 cell line. Methods Cell viability, apoptosis, and cell cycle were measured by Muse Cell Analyzer. All the protein levels were determined by the ELISA method. Results We observed that silibinin significantly reduced cell proliferation and colony formation (p<0.05, p<0.001, respectively). Silibinin also significantly induced total apoptosis and the G0/G1 phase of the cell cycle (p<0.01). We observed that silibinin significantly decreased JAK2 levels while increased STAT1 levels compared to the controls respectively (p<0.001; p<0.001). Besides, silibinin statistically decreased the levels of sAXL; however, numerically, but not statistically, it increased the level of GAS6 (p>0.05). Conclusions Silibinin reduces colony formation, inducing apoptosis and arresting cancer cells in the G0/G1 phase, which is an indicator of its anticancer activity. In addition, silibinin decreased the levels of JAK2 and sAXL, which contributes to the development of cancer, and increased the levels of STAT1, suggesting silibinin’s antiproliferative effects.

Objectives Dentin tissue can act as a reservoir for bioactive molecules that create signals for cellular proliferation and differentiation to initiate tissue regeneration. Therefore, the aim of this study is to compare the cell viability, inflammatory response, and antimicrobial activity of bovine dentin grain-added calcium-hydroxide (CH-BDG) with different pulp-capping materials. Methods ProRoot MTA, Biodentine, Dycal, TheraCal-LC, and an experimental material, CH-BDG, were examined. Cell viability was determined via the WST-1 assay. The inflammatory response was analysed by the monocyte chemoattractant protein-1 (MCP-1/CCL2) and macrophage inflammatory protein-1α (MIP-1α/CCL3) levels. The antimicrobial activity was evaluated by agar-diffusion. Results The cell viability of CH-BDG was analogous with Biodentine and control at 24 h. The cell viability of CH-BDG decreased at 48 h, but the rate was higher than ProRoot MTA and Dycal (p<0.05). For MCP-1 and MIP-1α values, there was no significant difference between the control and CH-BDG. The MCP-1 level of CH-BDG was lower compared to other pulp-capping materials (p<0.05). The MIP-1α level of CH-BDG was lower compared to ProRoot MTA, Biodentine, and TheraCal-LC (p<0.05). No inhibition zone was detected against oral microorganisms for CH-BDG. Conclusions The experimentally developed CH-BDG showed competing properties and additional advantages compared to the existing pulp-capping materials.

Objectives Curcumin plays a leading role as an epigenetic regulator in cancer. miR-15a-5p is a crucial non-coding RNA for breast cancer (BRCA) and various cancers due to its tumor suppressor role. In our study, we aimed to examine the curcumin/miR-15a-5p/target gene interaction in BRCA cells. Methods The effects of curcumin and miR-15a-5p on cell viability in the MCF7 cells were examined using the WST8 technique. The cell migration was determined using scratch wound assay. miR-15a-5p level was detected in curcumin-treated cells and miR-15a-5p transfected cells compared to control groups by RT-qPCR. Overexpressed genes in BRCA were found by bioinformatics tools (GSE41970 and TCGA). miR15a-5p potential target genes in the miRNet tool were selected in overlapped genes between GSE41970 and TCGA. Survival analysis of the selected genes was examined using the GEPIA2 tool. Relative expression levels of four selected genes were examined via qPCR. Results Cell viability and scratch-wound closure rate were reduced in curcumin-treated and miR-15a-5p mimic transfected MCF7 cells. miR15a-5p overexpressed in curcumin-treated and miR-15a-5p transfected cells. Eighty-three dysregulated upregulated genes were detected (in GSE41970 and TCGA). Among the possible target genes of miR-15a-5p in the miRNet tool, 10 upregulated genes were detected overlapping with GSE41970 and TCGA. CCNE1 and CHEK1 genes were found to be important for survival in BRCA. CCNE1 and BMI1 were decreased in curcumin-treated and miR-15a-5p transfected cells. Conclusions Curcumin treatment increased miR-15a-5p and downregulated selected target genes. Curcumin/miR-15a-5p interaction may be a much stronger negative regulator of the CCNE1 and BMI1 genes in BRCA.

Objectives To investigate the cellular and apoptotic effects of food additive calcium propionate by in vitro methods. Methods Cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, intracellular Glutathione (GSH) by luminometric method, mitochondrial membrane potential (MMP) by fluorometry, apoptosis by dual Acridine Orange/Ethidium Bromide (AO/EB) fluorescent staining were studied in lung cells. Results It has been observed that calcium propionate reduced GSH and MMP levels significantly while increased cytotoxicity, apoptosis, and reactive oxygen species (ROS) levels in both A549 and DMS114 cancer cells in a dose-dependent manner. Conclusions Cytotoxic effects of calcium propionate were more pronounced in cancer cells compared to healthy cells, suggesting its potential suitability as a chemo-preventive agent.

Objectives Lung cancer is a highly prevalent and life-threatening disease worldwide, with non-small cell lung cancer (NSLC) accounting for around 80 % of all cases. Exosomes contain important genetic information for humans that could be employed, especially for early screening of tumors. Accordingly, we aimed to use exosomal miRNA (ex-miRNA) in early NSCLC diagnosis. Methods The extracted ex-miRNAs were validated through transmission electron microscopy, particle size potentiometer, and western blot analyses. Microarray was used to verify ex-miRNAs, and 20 miRNAs were selected. Herein, we obtained 240 blood samples from NSCLC patients (101 in the early stage) and 234 from healthy donors. Our study deployed real-time fluorescence quantitative PCR (qRT-PCR) for detecting significantly down-regulated miR-587 expression. In addition, the hypersensitive C-reactive protein (Hs-CRP) levels were measured in patient samples. Results The results of calculating the area under the curve (AUC) revealed that the diagnostic efficiency of miR-587 and Hs-CRP were 0.771 and 0.863, respectively. Meanwhile, the combined diagnostic efficiency of both increased to 0.901. In patients with early NSCLC, the diagnostic efficiency of miR-587, Hs-CRP, and combined AUC were 0.726, 0.873, and 0.899, respectively. This indicates that the accuracy of early NSCLC diagnosis is very high. Finally, we combined miR-587 and Hs-CRP with CEA and NSE for NSCLC (AUC=0.956) and early-stage patients (AUC=0.921). Conclusions In this study, miR-587 and Hs-CRP have significant diagnostic efficiency for NSCLC, especially the combination of CEA and NSE that could indicate early NSCLC diagnosis.

Objectives Mutations detected in the AVPR2 gene (arginine vasopressin type 2 receptor) are known to cause nephrogenic diabetes insipidus (NDI). Several pharmacological chaperones (PCs) target misfolded AVPR2 proteins and rescue them from the quality control system of the cell. In this study, we investigated the effect of YM087 and VPA985, which are PCs, on T273M-AVPR2 mutant that are known to cause NDI. Methods The total and cell surface expressions of T273M in COS-1 cells were measured by sandwich ELISA and flow cytometry after the cells were treated with YM087 and VPA985 separately. In addition, the cAMP accumulation assay was performed. Results It was observed that VPA985 treatment significantly increased the cell surface expression and slightly increased the maximum cAMP response of T273M. Both YM087 and VPA985 decreased the ligand concentrations which were responsible for making half of the maximum response of the T273M mutant receptor. Conclusions PCs have different potential effects on different AVPR2 mutants. Therefore, studying the effectiveness of PCs in rescuing AVPR2 mutants and making them functional again may contribute to the development of new therapeutic strategies.

Objectives We aimed to investigate the relationship between serum paraoxonase-1 (PON1) and paraoxonase-3 (PON3) levels and activities with hemoglobin A1c (HbA 1c ), serum fasting blood glucose, lipid profile, and oxidative stress in patients with type 2 diabetes mellitus (T2DM). Also, we aimed to examine PON1 and PON3 levels and activities in these patients according to the HbA 1c goal in diabetes treatment and PON1 192 phenotypes. Methods One hundred forty-one volunteers diagnosed with T2DM participated in this study. Serum PON1 and PON3 levels and activities, total oxidant status (TOS), and total antioxidant status (TAS) were measured. PON1 192 phenotypes were determined by using PON1 activities. Also, HbA 1c , serum fasting blood glucose, and lipid profile results, which were measured for routine examination on the same day as sample collection, were used for this study. Results There was a positive relationship between arylesterase and lactonase activities and high-density lipoprotein cholesterol (HDL-C), between lactonase activity and TAS, and a negative relationship between PON1 level and TAS in patients with T2DM. Our study also showed that PON3/HDL-C was higher in patients with HbA 1c levels ≥7 %. Lactonase activities were higher in patients with PON1 Q192Q and PON1 Q192R phenotypes than in patients with PON1 R192R phenotypes. Conclusions PON1 and PON3 levels and activities alone could not be associated with immediate or long-term blood glucose levels in patients with T2DM. Higher PON3/HDL-C in patients with HbA 1c levels ≥7 % may show a protective role of PON3 in defense against higher glucose levels. Also, we found that the PON1 192 phenotype can affect serum lactonase activity.

Objectives BLM , a member of the RecQ helicase family, plays an important role in DNA repair, and its biallelic mutations cause autosomal recessive Bloom syndrome, a disease characterized by elevated levels of sister chromatid exchange (SCE) in affected individuals and hereditary cancer susceptibility in carriers. This study aims to investigate genomic instability in breast cancer patients carrying heterozygous variants in the BLM gene. Methods Spontaneous chromosome breakage count and SCE counting were performed on newly drawn blood cultures, both spontaneous and stimulated. The spontaneous breakage count was conducted alongside control samples. In SCE analysis, 0–10 per metaphase was considered normal, 10–40 borderline, and counts above 40 were considered high. Results The study included 26 patients and one healthy control at each session. The clinical and pathological characteristics of the patients were evaluated. The analyses revealed borderline-level increased SCE rates in only one patient. No increase in spontaneous breakage count or SCE analysis was observed in other individuals compared to controls. Conclusions Increased genomic instability was not observed in the analyzed patient group. These results can lead to multiple interpretations. The variants carried in the BLM gene in the patient group may be of low pathogenicity, or increased instability compared to controls may not be necessary for heterozygous variants. Additionally, our patient group may not have been exposed to a genotoxic effect causing genomic instability. These results could also indicate a favorable position in terms of avoiding chemotherapy and radiotherapy complications.

Objectives The presence of live SARS-CoV-2 viruses in the fecal specimens and the positive results for SARS-CoV-2 RNA in gastrointestinal samples after respiratory specimens had become negative indicate that there may be a risk of transmission of the disease not only through the respiratory tract but also through the fecal-oral route. The study aim is to determine the time period that the SARS-CoV-2 virus remains in the fecal specimens of discharged COVID-19 patients to reveal the time interval in which the risk of transmission continues. Methods In 65 patients hospitalized with a COVID-19 diagnosis, the viral RNA was isolated from the supernatant of the stool sample using CVXTM viral RNA extraction kit. The extracted RNAs from the stool samples were detected using a commercial RT-PCR method. Results Positive results were obtained in the stool samples in eight of sixty-five patients and the presence of SARS-CoV-2 in these patients was detected for up to three weeks. Conclusions By determining the residence time of SARS-CoV-2 virus in stool samples of discharged COVID-19 patients, the time interval during which the possibility of transmission risk continues has been revealed. The findings of the study could prove beneficial in comprehending the risks of transmission and translating them into preventative measures.