Boron-Doped Carbon Nanodots as a Theranostic Agent for Colon Cancer Stem Cells.

Carbon nanodots have drawn a great deal of attention due to their green and expedient opportunities in biological and chemical sciences. Their high fluorescence capabilities and low toxicity for living cells and tissues make them excellent imaging agents. In addition, they have a fluorimetric response against inorganic and organic species. Boron-doped carbon nanodots (B-CDs) with high fluorescence yield were produced from phenylboronic acid and glutamine as boron and carbon sources, respectively, by a hydrothermal method. First, the effects of the temperature on their fluorescence yield and the structural characteristics of B-CDs were investigated. Second, their cytotoxicity and cell death and proliferation behaviors were examined. The cytotoxicity was evaluated by the MTT assay. The cellular properties were evaluated with the distribution of caspase 3, Ki67, lamin B1, P16, and cytochrome c after the indirect immunoperoxidase technique. After the MTT assay, 1:1 dilution of all applicants for 24 h was used in the study. After immunohistochemical analyses, the application of B-CDs synthesized at 230 °C did not change control cell (Vero) proliferation, and also apoptosis was not triggered. Colo 320 CD133+ and CD133- cell-triggered apoptosis and cellular senescence were found to be synthesis temperature dependent. In addition, Colo 320 CD133- cells were affected relatively more than CD133+ cells from B-CDs. While B-CDs did not affect the control cells, the colon cancer stem cells (Colo 320 CD133+) were affected in a time-dependent manner. Therefore, the use of the synthesized B-CD product may be an alternative method for controlling or eliminating cancer stem cells in the tumor tissue.

SnO2 nanoparticles/waste masks carbon hybrid materials for DNA biosensor application on voltammetric detection of anti-cancer drug pazopanib.

This present study is the first investigation of pazopanib-dsDNA binding using bare and modified GCE. The interaction was mainly evaluated based on the decrease of voltammetric signal of deoxyadenosine by differential pulse voltammetry using three different ways, including the incubated solutions, dsDNA biosensor, and nanobiosensor. The nanobiosensor was fabricated with the help of SnO2 nanoparticles and carbon hybrid material. The carbon material is derived from the waste mask, the most used personal protective equipment for the ongoing COVID-19 pandemic. Both materials were synthesized via the green synthesis technique and characterized by various techniques, including BET, TEM, SEM-EDX, AFM, XPS, and XRD. Spectrophotometric and molecular docking studies also evaluated the pazopanib-dsDNA binding. All calculations showed that pazopanib (PZB) was active in the minor grove region of DNA.

Human hair rich in pyridinic nitrogen-base DNA biosensor for direct electrochemical monitoring of palbociclib-DNA interaction.

Carbon material derived from the waste-based biomass human hair (H), which is naturally rich in pyridinic nitrogen, provides a significant benefit in biosensor applications with its dominant conductivity character. The carbon material was synthesized from human hair waste by the hydrothermal carbonization (HTC) method, which is a promising green synthesis. A morphological characterization of the carbon materials was performed. In this study, H and amine-functionalized multi-walled carbon nanotubes (NH2-MWCNT) were combined for the first time as a modifier, which enhanced the glassy carbon electrode (GCE) surface area for deoxyribonucleic acid (DNA) biosensor studies. Palbociclib (PLB) is clinically used in the treatment of breast cancer. The novel electrochemical nanobiosensor was used to investigate the dsDNA-PLB interaction to evaluate the possibility that PLB causes conformational changes in DNA structure and/or oxidative damage. The interaction was conducted based on the voltammetric signals of deoxyguanosine (dGuo) and deoxyadenosine (dAdo) by differential pulse voltammetry (DPV) on a bare and H + NH2-MWCNT modified GCE. The proposed analytical method was applied to a pharmaceutical dosage form with a satisfactory recovery of 98.25 %. The nanobiosensor was tested in the presence of some interfering agents. The binding mechanism of dsDNA-PLB was also evaluated by spectroscopic and theoretical calculations.

Recent biopharmaceutical applications of capillary electrophoresis methods on recombinant DNA technology-based products.

Biopharmaceuticals (recombinant technology-based products, vaccines, whole blood and blood components, gene therapy, cells, tissues, etc.,) are described as biological medical products produced from various living sources such as human, microbial, animal, and so on by manufacturing, extraction, or semi-synthesis. They are complex molecules having high molecular weights. For their safety and efficacy, their structural, clinical, physicochemical, and chemical features must be carefully controlled, and they must be well characterized by analytical techniques before the approval of the final product. Capillary electrophoresis (CE) having versatile modes can provide valuable safety and efficacy information, such as amino acid sequence, size variants (low and high molecular weight variants), charged variants (acidic and basic impurities), aggregates, N-linked glycosylation, and O-linked glycosylation. There are numerous applications of CE in the literature. In this review, the most significant and recent studies on the analysis of recombinant DNA technology-based products using different CE modes in the last ten years have been overviewed. It was seen that the researches mostly focus on the analysis of mAbs and IgG. In addition, in recent years, researchers have started to prefer CE combined mass spectrometry (MS) techniques to provide a more detailed characterization for protein and peptide fragments.

Replacement of antibodies with bacteriophages in lateral flow assay of Salmonella Enteritidis.

In this study, the analytical performance of bacteriophages for Salmonella Enteritidis was investigated using lateral flow assay (LFA) technique. The analytical performance characteristics of bacteriophages were compared with antibodies which are regularly used as analyte-specific agents in the lateral flow immunoassay test strip. Bacteriophages could be an alternative analyte-specific agents to antibodies in lateral flow assay testing of bacteria since they offer comparable sensitivity, specificity, and accuracy. In the present study, Surface Enhanced Raman Spectroscopy (SERS) and colorimetric measurements were combined in one platform and sensitive quantitation of target bacteria was accomplished with a total quantitative analysis time of less than 30 min. The developed Salmonella Enteritidis F5-4 phage-based LFA specifically responds to Salmonella Enteritidis, while lower SERS responses to different bacteria types including Bacillus subtilis, Micrococcus luteus, Escherichia coli, Salmonella Typhimurium were observed. The developed test strips were also applied for the determination of Salmonella Enteritidis in spiked chicken and egg samples.

Electrochemical, spectroscopic, and molecular docking studies of the interaction between the anti-retroviral drug indinavir and dsDNA.

In this study, an electrochemical DNA biosensor was developed using a straightforward methodology to investigate the interaction of indinavir with calf thymus double-stranded deoxyribonucleic acid (ct-dsDNA) for the first time. The decrease in the oxidation signals of deoxyguanosine (dGuo) and deoxyadenosine (dAdo), measured by differential pulse voltammetry, upon incubation with different concentrations of indinavir can be attributed to the binding mode of indinavir to ct-dsDNA. The currents of the dGuo and dAdo peaks decreased linearly with the concentration of indinavir in the range of 1.0-10.0 µg/mL. The limit of detection and limit of quantification for indinavir were 0.29 and 0.98 µg/mL, respectively, based on the dGuo signal, and 0.23 and 0.78 µg/mL, respectively, based on the dAdo signal. To gain further insights into the interaction mechanism between indinavir and ct-dsDNA, spectroscopic measurements and molecular docking simulations were performed. The binding constant (Kb) between indinavir and ct-dsDNA was calculated to be 1.64 × 108 M-1, based on spectrofluorometric measurements. The obtained results can offer insights into the inhibitory activity of indinavir, which could help to broaden its applications. That is, indinavir can be used to inhibit other mechanisms and/or hallmarks of viral diseases.

Fluorescence chemosensing of meldonium using a cross-reactive sensor array.

In this paper, we report a fluorescent sensor array approach for the urinary detection of a prohibited substance in sports, meldonium. Four chemosensors with ethidium bromide scaffolds were employed in this method. The interaction between meldonium and chemosensors was investigated by different techniques, such as ultraviolet-visible absorption and fluorescence spectroscopy, nuclear magnetic resonance, and mass spectrometry. Molecular dynamics simulation was also used to elucidate and support the interaction mechanisms between meldonium and the chemosensors. Differential responses obtained from the sensor array enabled the qualitative and quantitative analyses of meldonium with low error values. This method was able to detect and quantify meldonium at the nM level, fulfilling the requirements of minimum performance defined by the World Anti-Doping Agency.

Assessment of hand functions in patients with idiopathic cervical dystonia.

Cervical dystonia (CD) is the most common form of focal dystonia characterized by involuntary contractions of the neck muscles, causing abnormal rotation of the head into specific directions. Studies report that idiopathic dystonia is a developmental disorder of the sensorimotor circuits, involving both the cortico-striatal and thalamo-cortical pathways. It is also suggested that enhanced cortical plasticity extends beyond the clinically affected region and may also be detected in the unaffected upper limbs of the patient with CD. In the present study, we aimed at exploring if patients with CD had hand motor dysfunctions. Forty patients with idiopathic CD and 40 healthy controls were included in this study. Dystonic symptoms were assessed by means of The Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS). Stanford Health Assessment Questionnaire (HAQ) was used to assess functional status. Quality of life (QoL) was assessed by using the Medical Outcomes Study Short Form 36-Item Health Survey (SF 36). Grip strength was assessed by using hand dynamometers. Tip pinch, lateral pinch and chuck pinch of the hand were assessed by using a pinchmeter. Fingertip dexterity and hand coordination was assessed using Purdue Pegboard. Duruoz Hand Index (DHI) was used for the assessment of hand functions. There were no significant differences between the groups in grip and pinch strengths of hands and fingers. As to the fingertip dexterity, patients with CD had a mean Pin 1 and Pin 2 test score of 10.6 ± 2.8 and 10.8 ± 3.2 respectively and a mean assembling test score of 5.2 ± 2.0. These results were significantly worse than those of the healthy controls. As to the SF 36 sub-scores, there were significant differences between the groups in all SF 36 sub-scores (p < .001). This study indicates that patients with CD suffer a deteriorated fine motor coordination of hands without dystonic involvement of upper extremities. Furthermore, lower SF 36 scores in patients with CD suggest poorer health-related quality of life.

A Case of Refractory Polymyositis Successfully Treated With Abatacept Monotherapy.

Polymyositis (PM) is an autoimmune disease progressing in the form of a break down of the muscles that is induced by chronic inflammation in skeletal muscles. Muscle weakness is painless and concentrates on proximal muscles, involving the pectoral and pelvic girdle. If the disease is not treated properly, it may progress and lead to a considerable decrease in the quality of life. Its conventional treatment involves drugs that suppress inflammation such as steroids, methotrexate, azathioprine, and intravenous immunoglobulin. However, conventional treatment may prove insufficient to halt the progression of the disease and offer only a limited improvement because of the adverse effects it causes in some patients. In this article, we present a 48-year-old female patient diagnosed with PM nearly 13 years ago that did not sufficiently respond to the pharmaceutical agents that were indicated for the conventional treatment of the disease and developed femoral head avascular necrosis because of the treatment and was, in the end, successfully treated with abatacept monotherapy at our clinic.

Electrochemical, spectroscopic and molecular docking studies on the interaction of calcium channel blockers with dsDNA.

This study presents evaluation of the possible interaction mechanism between calf thymus dsDNA and three calcium antagonists; nifedipine, lercanidipine and amlodipine. The interactions between Nifedipine-dsDNA and Lercanidipine-dsDNA were investigated by differential pulse voltammetry using two different interaction methods; at the dsDNA-electrochemical biosensor surface and in bulk incubated solution. Amlodipine was used as model drug in bulk incubated solution. The decrease in the peak current of guanine and adenine were used as an indicator for confirmation of the interaction event in acetate buffer of pH 4.70. In bulk incubated solution, after interaction with Nifedipine and Amlodipine the guanine signal was almost disappeared. At the dsDNA modified glassy carbon electrode surface, the peak currents of guanine and adenine were decreased while Nifedipine and Lercanidipine interacts with DNA. The interactions between Nifedipine-dsDNA and Lercanidipine-dsDNA were further studied by UV-Vis absorption spectroscopy which indicates the intermolecular interaction between these drugs and ds-DNA can be mainly through hydrogen bonding and van der Waals forces. Molecular docking calculations shown that the AMP-1-2, NDP and LDP-1-2-ctDNA having groove binding. Beside spectral data, docking studies elicited that AMP-1-2, NDP and LDP-1-2 complexes have different interaction and conformation trends to target (ctDNA).

A New Comorbidity in Female Patients With Ankylosing Spondylitis: Pelvic Organ Prolapse.

AIM: The aim of this study was to investigate whether increased intra-abdominal pressure caused by pelvic inflammation and frequent use of the Valsalva maneuver increases the incidence of pelvic organ prolapse (POP) among female patients with ankylosing spondylitis (AS). METHODS: Thirty-nine patients diagnosed as having AS through the use of the modified New York criteria, 47 patients with chronic low-back pain (CLBP), and 38 healthy controls (HCs) were included in this study. All the patients and the HCs underwent thorough physical and gynecological examinations. Pelvic organ prolapse was graded blindly by a gynecologist. Presence or absence of cystocele (CS), rectocele (RC), and uterine prolapse (UP) was noted. RESULTS: The incidences of CS, RC, and UD were significantly higher among the AS patients compared with the HCs (p = 0.001, 0.026, and 0002, respectively). The incidences of CS (p = 0.042) and UD (p = 0.017) were significantly higher among the AS patients compared with the CLBP patients. CONCLUSIONS: The incidence of POP is higher among patients with AS compared with normal population. These patients should be questioned about the symptoms of POP and recommended, if necessary, regular gynecological examinations, as well as specific exercises including those targeting AS.

A useful marker in the assessment of remission and activation of disease in patients with rheumatoid arthritis: Serum human neutrophil peptides 1-3.

AIM: Rheumatoid arthritis (RA) is a chronic disease of unknown etiology. Various cellular and molecular immunological factors are involved in the pathophysiology of RA. Recent studies suggest that neutrophils and alpha-defensins released from the neutrophils assume significant roles in the pathogenesis of RA. The aim of this study was to investigate the potential association between serum alpha-defensin levels and disease activity, functional status, radiological damage and several laboratory parameters in patients with RA. MATERIALS AND METHODS: A total of 42 patients with established RA who presented to the outpatient clinics of rheumatology of Dicle University Hospital and 38 healthy control subjects were included in this study. Disease activity was assessed by using the Disease Activity Scale 28 (DAS28). Quality of life was assessed by using the Rheumatoid Arthritis Quality of Life (RAQoL) Questionnaire and the Nottingham Health Profile (NHP). Functional status was assessed by using the Stanford Health Assessment Questionnaire (HAQ). Laboratory examinations included the following tests: CBC, ESH, CRP, and HNP 1-3. RESULTS: Patients with an active disease exhibited higher HNP 1-3 levels compared to patients in remission. At a cut off value of 708 pg/ml, sensitivity and specificity of the tests for HNP 1-3 were 72% and 70.6%, respectively. CONCLUSION: In the present study, patients with an active disease had significantly higher serum HNP 1-3 levels compared to patients in remission. In this respect, serum HNP 1-3 can be a useful marker in the assessment of disease activity and remission in patients with RA.

Simultaneous determination and validation of emtricitabine, rilpivirine and tenofovir from biological samples using LC and CE methods.

A combination of antiretroviral agents is frequently used in effective treatment of the human immunodeficiency virus infection. In this study, two different separation methods are presented for the simultaneous determination of emtricitabine, rilpivirine and tenofovir from raw materials and urine samples. Developed liquid chromatography and capillary electrophoresis methods were thoroughly optimized for high analytical performances. Optimization of multiple variables at the same time by performing a minimum number of experiments was achieved by the Box-Behnken design, which is an experimental design in response surface methodology, in capillary electrophoresis. The results of the experimental design ensure minimum analysis time with well-separated analytes. Separation conditions, such as different stationary phases, pH level, organic modifiers and temperatures in liquid chromatography method, were also optimized. In particular, among stationary phases, the core-shell column especially enhanced the effectiveness of separation in liquid chromatography. Both methods were fully validated and applied to real samples. The main advantage of the developed methods is the separation of the drug combination in a short time with high efficiency and without any time-consuming steps.

Psychological health of caregivers and association with functional status of stroke patients.

OBJECTIVE: Stroke does not only affect the physical state of patients but also the emotional state of their relatives, most effectively their caregivers. The study aims to examine the mood of caregivers experienced with care for patients with stroke who are highly dependent on the assistance and also to establish the relationship between the emotional state of caregivers and the severity of disability of the patients. METHODS: This study contained a total of 76 patients with sufficient cognitive functions and severe physical disabilities with hemiplegia caused by a cerebrovascular accident and their caregivers and 94 controls. The functional state of patients was assessed by the Barthel Index (BI). Furthermore, emotional state of the caregivers was assessed by the Hospital Anxiety and Depression Scale (HADS) and their life quality was assessed by the SF36 Health Survey. RESULTS: The mean anxiety (9.73 ± 4.88) and depression rates (9.81 ± 5.05) in the caregivers were significantly higher than those in controls (p<0.001, respectively). Significant impairments were observed in both their mental and physical health. Regression analysis also showed a significant negative correlation between the BI scores and the HADS scores. CONCLUSION: Caregivers had an impaired emotional state and the level of their anxiety was associated with the severity of functional disability of the patients. Therefore, the support provided to the caregiver might be influential on the functional recovery of the patients.

The Effects of 1,3,5-trisubstituted Indole Derivatives on Cell Growth, Apoptosis and MMP-2/9 mRNA Expression of MCF-7 Human Breast Cancer Cells.

BACKGROUND: Matrix metalloproteinases are known as extracellular matrix degrading enzymes and have important role on tumor progression. OBJECTIVE: This study reports the effects of 1,3,5-trisubstituted indole derivatives on cytotoxicity, apoptosis and MMP- 2/MMP-9 mRNA expression of MCF-7 human breast carcinoma cells. METHOD: The cytotoxic effects of the compounds on MCF-7 cells were performed by MTT test, and cell proliferation was determined via BrdU incorporation. The apoptotic effects were observed by cell death detection elisa. The effects of the compounds on MMP-2/-9 enzyme activity and mRNA expression were also performed. RESULTS: The compounds inhibited the proliferation of MCF-7 breast carcinoma cells significantly in a dose dependent manner. All compounds were able to induce DNA fragmentation, especially compound 1. The IC50 values of compound 2 and 4 for MMP-2 were 0.42 µM and 1.88 µM, respectively. MMP-2 mRNA expression results were correlated with the inhibition of enzyme activity, such compound 4 inhibited MMP-2 mRNA expression at all treated concentrations. Docking simulation has also been performed to analyze the binding mode of compounds and the results showed that compound 2, the most active compound, formed a hydrogen bond with Glu202 for binding to the MMP-2 active site. In addition, the hydrophobic parts of compound 2 are in contact with nonpolar surface areas of MMP-2, such as His201, His211, Tyr223 and Tyr193. CONCLUSION: According to the molecular docking results along with the biological assay data, it is suggested that compound 2 might be used for further design and development of MMP-2 inhibitors.

Surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin in urine.

We present a surface-enhanced Raman probe (SERS) platform for the determination of a prohibited substance, recombinant erythropoietin (rEPO), in urine matrix, using nanoparticles as substrate. Rod-shaped gold nanoparticles (GNR) were modified with a Raman label and an antibody as SERS probe. We developed two SERS-based immunoassays for detection and quantification of rEPO in urine. In the first assay, rEPO was determined by a sandwich assay with gold surfaces and GNR. In the second assay, rEPO was extracted by using core shell-structured magnetic iron oxide gold nanoparticles, and again sandwich assay was performed by using GNR. We also demonstrated the ability of the proposed method to discriminate rEPO and urinary erythropoietin (uEPO). A good linear correlation was obtained between logarithms of rEPO concentrations in urine and Raman intensities within the range of 10-1-103 pg mL-1 rEPO concentrations. Detection limits which are smaller than 0.1 pg mL-1 levels were achieved owing to the high extractive performance of the nanoextraction techniques. Graphical Abstract Schematic represantation of surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin.

Detection and quantification of ATP in human blood serum.

Two fluorometric sensors based on the tri-serine tri-lactone scaffold and thiourea or sulfonamide moieties serving as hydrogen bond donors allowing for anion binding are described. The sensor utilizing thiourea as a recognition moiety shows fluorescence enhancement while the sensor with sulfonamide shows quenching upon addition of phosphates. Sensor arrays composed of two sensors are able to discriminate structurally similar organic phosphates in the presence of interferents in human blood serum. The quantitative analysis of ATP in human blood serum shows high accuracy (the root mean square error of prediction, 1.65%) without requiring any sample pretreatment.

Fluorescent zinc and copper complexes for detection of adrafinil in paper-based microfluidic devices.

Recognition of electroneutral Lewis bases and anions in aqueous media is extremely difficult. We show that fluorescent coordinatively unsaturated metal complexes can recognize various Lewis bases while providing an easy-to-detect fluorescence response. This approach is applied to the detection of adrafinil, a banned performance-enhancing drug.

A tri-serine tri-lactone scaffold for the quantification of citrate in urine.

Tri-serine tri-lactone based C3 symmetry fluorescent sensors were synthesized. Citrate is shown to bind to sensors, while displaying an increase in fluorescence intensity for the sensor with thiourea and a quenching for the sensor with sulfonamide. Information-rich responses of the sensors enable us to discriminate structurally similar anions, including mono-, di- and tri-carboxylates with 100% correct classification. A simple two-sensor array enables the determination of the concentration of citrate in urine without any sample preparation with high accuracy (error < 2%).

Gold nanoparticle-lignan complexes inhibited MCF-7 cell proliferation in vitro: a novel conjugation for cancer therapy.

Nanoparticles, including gold nanoparticles (AuNP), have been used in imaging in cancer treatment and as therapeutic agents and drug delivery vehicles. Particularly lignans, also called phytoestrogens, have strong effects on the treatment of carcinomas due to their antiestrogenic, antiangiogenic and proapoptotic mechanism. The aim of this study is to investigate the antiproliferative effects of three lignans-AuNP conjugates, pinoresinol (PINO), lariciresinol (LARI) and secoisolariciresinol (SECO), on the MCF-7 cell lines. For this purpose, first, thiolated ß-cyclodextrin (ß-CD) was synthesized to achieve a surface modification of AuNP, and then the ß-CD modified AuNP was characterized using the transmission electron microscopy (TEM), UV-Visible and Nuclear Magnetic Resonance (NMR) spectroscopy. Then, the selected lignans were conjugated to the ß-CD-modified AuNP, and the antiproliferative effect of these conjugates was monitored. The results suggest that when compared to their non-conjugated forms, the AuNP-bound lignan conjugates prevented the proliferation of the MCF-7 cells significantly. Therefore, these AuNP-conjugated derivatives can be new candidate agents for breast cancer therapy.