Ultrastructural Changes in the Ovariole of Isophya nervosa Ramme, 1931 (Orthoptera: Tettigoniidae) and Egg Morphology.

This study was conducted to assess the morphology of eggs and histology of the ovaries in female Isophya nervosa Ramme, 1931 (Orthoptera: Tettigoniidae). While the egg morphology of I. nervosa was studied and examined by a stereomicroscope, a light microscope, and a scanning electron microscope, respectively, the morphology and histology of the ovary of this species were studied and examined by a stereomicroscope, a light microscope, a scanning electron microscope, and a transmission electron microscope, respectively. We found that the adult female had two pairs of ovaries, lateral oviduct, common oviduct, and spermatheca. Morphological study of the ovariole revealed that it is categorized under panoistic type of ovariole which is divided into three regions, the terminal filament, the germarium, and the vitellarium. We also observed that the eggs in I. nervosa have an ellipsoidal shape and are brown in color. Three different layers such as extrachorion, exochorion, and endochorion were observed. When the egg morphology is examined, it is understood that the surface pattern of the egg and the features of the micropylar areas may be distinguishing characters at the subfamily level, in addition to known classical taxonomic characters.

Silencing of an ABC transporter, but not a cadherin, decreases the susceptibility of Colorado potato beetle larvae to Bacillus thuringiensis ssp. tenebrionis Cry3Aa toxin.

The Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae), is a major pest of potato plants worldwide and is notorious for its ability to develop resistance to insecticides. Cry3 toxins synthesized by Bacillus thuringiensis ssp. tenebrionis have been used successfully to manage this pest. Resistance to Cry toxins is a concerning problem for many insect pests; therefore, it is important to determine the mechanisms by which insects acquire resistance to these toxins. Cadherin-like and ABC transporter proteins have been implicated in the mode of action of Cry toxins as mutations in these genes render lepidopterans resistant to them; however, clear consensus does not exist on whether these proteins also play a role in Cry3 toxin activity and/or development of resistance in coleopterans. In the current study, we identified the L. decemlineata orthologues of the cadherin (LdCAD) and the ABCB transporter (LdABCB1) that have been implicated in the mode of action of Cry toxins in other coleopterans. Suppression of LdABCB1 via RNA interference reduced toxin-related larval mortality, whereas partial silencing of LdCAD did not. Our results suggest that the ABCB is involved in the mode of action of Cry3Aa toxins; however, no evidence was found to support the role of cadherin as a receptor of Cry3Aa in L. decemlineata.

The Ovariol Morphology and Ultrastructure of Poecilimon ataturki Ünal, 1999 (Orthoptera, Tettigoniidae) and the Histochemical Features of the Yolk Granules.

This study presents the oocyte development of Poecilimon ataturki Ünal, 1999 (Orthoptera, Tettigoniidae) with histology, morphology, and histochemistry by using a stereomicroscope, a light microscope, a scanning electron microscope, and a transmission electron microscope. The ovary in this species is a panoistic type which contains many ovarioles which consist of terminal filament, germarium, and vitellarium. Germarium is the region that has undifferentiated cells which generate the oocytes and follicular cells. In the vitellarium region, yolk granules start to cover the whole oocyte. In histochemical studies, to determine the content of the yolk granules, proteins, and carbohydrates in oocytes were treated with a bromophenol blue (BPB) method, a mercury bromophenol blue (mBPB) method, and a periodic acid Schiff (PAS) method, respectively. As a result of these methods, the yolk granules gave positive results in ovariole sections treated with the PAS and the BPB, while the mBPB staining was negative.

Histomorphology of the Malpighian Tubules and the Chemical Composition of the Spherocrystals in the Tubule Epithelial Cells of Adult Leptophyes albovittata (Kollar, 1833) (Orthoptera, Tettigoniidae).

In insects, the number, cytological and histological structures, and the spherocrystals of the Malpighian tubules (MTs) can vary considerably in different insect groups. These differences are considered important because they can be used as taxonomic characters. For this purpose, the ultrastructure of the MT epithelial cells in Leptophyes albovittata (Kollar, 1833) (Orthoptera, Tettigoniidae) was examined by light microscopy, scanning electron microscopy, and transmission electron microscopy. The wall of each tubule consists of a single layer of cells. These cells have round-shaped nuclei. Two different cell types were demonstrated in the tubule cell. These are cells that have electron-dense cytoplasm and electron-lucent cytoplasm. It was observed that the cytoplasm of these cells has many spherocrystals. The chemical composition of the spherocrystals was found to be high in carbon, phosphorus, and manganese in tubule cells.

Immunomagnetic separation and Listeriamonocytogenes detection with surface-enhanced Raman scattering

Background/aim: We aimed to develop a rapid method to enumerate Listeria monocytogenes (L. monocytogenes) utilizing magnetic nanoparticle based preconcentration and surface-enhanced Raman spectroscopy measurements. Materials and methods: Biological activities of magnetic Au-nanoparticles have been observed to have the high biocompatibility, and a sample immunosensor model has been designed to use avidin attached Au-nanoparticles for L. monocytogenes detection. Staphylococcus aureus (S. aureus) and Salmonella typhimurium (S. typhimurium) bacteria cultures were chosen for control studies. Antimicrobial activity studies have been done to identify bio-compatibility and bio-characterization of the Au-nanoparticles in our previous study and capturing efficiencies to bacterial surfaces have been also investigated. Results: We constructed the calibration graphs in various population density of L. monocytogenes as 2.2 × 101 to 2.2 × 106 cfu/mL and the capture efficiency was found to be 75%. After the optimization procedures, population density of L. monocytogenes and Raman signal intensity showed a good linear correlation (R2 = 0.991) between 102 to 106 cfu/mL L. monocytogenes. The presented sandwich assay provides low detection limits and limit of quantification as 12 cfu/mL and 37 cfu/mL, respectively. We also compared the experimental results with reference plate-counting methods and the practical utility of the proposed assay is demonstrated using milk samples. Conclusion: It is focused on the enumeration of L. monocytogenes in milk samples and the comparision of results of milk analysis obtained by the proposed SERS method and by plate counting method stay in food agreement. In the present study, all parameters were optimized to select SERS-based immunoassay method for L. monocytogenes bacteria to ensure LOD, selectivity, precision and repeatablity.

Tracing Size and Surface Chemistry-Dependent Endosomal Uptake of Gold Nanoparticles Using Surface-Enhanced Raman Scattering.

Surface-enhanced Raman scattering (SERS)-based single-cell analysis is an emerging approach to obtain molecular level information from molecular dynamics in a living cell. In this study, endosomal biochemical dynamics was investigated based on size and surface chemistry-dependent uptake of gold nanoparticles (AuNPs) on single cells over time using SERS. MDA-MB-231 breast cancer cells were exposed to 13 and 50 nm AuNPs and their polyadenine oligonucleotide-modified forms by controlling the order and combination of AuNPs. The average spectra obtained from 20 single cells were analyzed to study the nature of the biochemical species or processes taking place on the AuNP surfaces. The spectral changes, especially from proteins and lipids of endosomal vesicles, were observed depending on the size, surface chemistry, and combination as well as the duration of the AuNP treatment. The results demonstrate that SERS spectra are sensitive to trace biochemical changes not only the size, surface chemistry, and aggregation status of AuNPs but also the endosomal maturation steps over time, which can be simple and fast way for understanding the AuNP behavior in single cell and useful for the assisting and controlling of AuNP-based gene or drug delivery applications.

SERS-based rapid assay for sensitive detection of Group A Streptococcus by evaluation of the swab sampling technique.

Beta-hemolytic, Group A Streptococcus pyogenes (GAS) is a life-threating pathogen and the reason for prominent disease, pharyngitis. The conventional analysis of GAS, gold standard, takes 48 hours and the related rapid tests lack in accuracy and sensitivity. In this study, firstly, the efficiency of swab sampling, which is a must in the GAS detection, was discussed with the proposed surface-enhanced Raman spectroscopy (SERS)-based batch assay and each step was controlled by the plate-counting method. Secondly, SERS-based lateral flow immunoassay (LFIA) test strips were constructed and the variation in the SERS intensity of 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) was observed. Thus, a linear correlation was found with a R2 value of 0.9926 and the LOD was calculated to be 0.2 CFU mL-1 of GAS which could be counted as one cell. The combination of the gold standard with the LFIA-SERS technique enabled the fast and accurate pathogen detection. In addition, GAS was quantified with paper-based test strips up to 100 CFU ml-1 level of bacteria for the first time without any interference. Besides, this study was featured with the discussion of the whole cell and pretreated cell detection of pathogens with LFIAs. Therefore, this work enlightens the points that have never been discussed on pathogen detection with paper-based platforms.

Tamoxifen-Related Thrombosis: An Experimental Study in Rat Venous Microvascular Anastomosis Model.

Tamoxifen is an estrogen receptor modulator and has been shown to increase risk for microvascular flap complications. This study aimed to investigate the clinical and histopathological effects of tamoxifen use in venous microvascular anastomosis model in rats. The role of vitamin E combination therapy and discontinuing tamoxifen therapy preoperatively were also evaluated.Forty rats were equally divided into 4 groups as follows: group 1 was given saline by oral gavage, group 2 was given tamoxifen citrate, group 3 was given tamoxifen citrate and vitamin E, and in group 4, tamoxifen citrate was given everyday except between days 12 and 16. In each group, femoral veins were dissected in each side and end-to-end anastomosis was performed in one side. Clinical and histopathological evaluations were performed. The ratio of total endothelial area to total vein area in a cross-sectional view of the vein was evaluated and compared.All veins with anastomosis in postoperative 15 minutes were found to be patent. In postoperative 1 week in groups 1 to 4, visible thrombus were present in 1, 3, 2, and 3 samples, respectively. Vitamin E group showed similar histopathological findings with control group. The ratio of endothelial layer to total vein cross-sectional area was increased in groups 2 and 4 in all samples. The increase was statistically significant between groups 2NA and 3NA (P = 0.023) and 2A and 1A (P = 0.006).Chronic tamoxifen consumption in the presence of anastomosis have led to prominent endothelial proliferation in rat femoral veins. Vitamin E combination therapy reversed this endothelial proliferation and should be focused in future studies.

Paper membrane-based SERS platform for the determination of glucose in blood samples.

In this report, we present a paper membrane-based surface-enhanced Raman scattering (SERS) platform for the determination of blood glucose level using a nitrocellulose membrane as substrate paper, and the microfluidic channel was simply constructed by wax-printing method. The rod-shaped gold nanorod particles were modified with 4-mercaptophenylboronic acid (4-MBA) and 1-decanethiol (1-DT) molecules and used as embedded SERS probe for paper-based microfluidics. The SERS measurement area was simply constructed by dropping gold nanoparticles on nitrocellulose membrane, and the blood sample was dropped on the membrane hydrophilic channel. While the blood cells and proteins were held on nitrocellulose membrane, glucose molecules were moved through the channel toward the SERS measurement area. Scanning electron microscopy (SEM) was used to confirm the effective separation of blood matrix, and total analysis is completed in 5 min. In SERS measurements, the intensity of the band at 1070 cm(-1) which is attributed to B-OH vibration decreased depending on the rise in glucose concentration in the blood sample. The glucose concentration was found to be 5.43 ± 0.51 mM in the reference blood sample by using a calibration equation, and the certified value for glucose was 6.17 ± 0.11 mM. The recovery of the glucose in the reference blood sample was about 88 %. According to these results, the developed paper-based microfluidic SERS platform has been found to be suitable for use for the detection of glucose in blood samples without any pretreatment procedure. We believe that paper-based microfluidic systems may provide a wide field of usage for paper-based applications.

Rare eggshell structure in Odonata: Lindenia tetraphylla (Van der Linden) (Anisoptera: Gomphidae).

Lindenia tetraphylla (Van der Linden, 1825) eggs exhibit an egg structure that is very rare in other Gomphidae species. They have a well-developed surface reticulation structure. The anterior pole of the egg has a small, rounded micropylar area consisting of seven orifices arranged radially around a central area. The posterior pole has a sessile, truncated cone that carries 55-65 coiled filaments. The filament structure found at the posterior pole of the egg has been observed in the gomphid species Lestinogomphus africanus (Fraser, 1926), Ictinogomphus australis (Selys, 1873), and I. rapax (Rambur, 1842). However, L. tetraphylla eggs differ from these species in both morphology and filament structure. This study provides a detailed analysis of the ultrastructure of L. tetraphylla eggs using scanning electron microscopy, and the functional and taxonomic significance of the eggshell are discussed. RESEARCH HIGHLIGHTS: The aim of this study is to examine the ultrastructure of the L. tetraphylla eggshell, emphasizing its function and taxonomic value. In this context, the general morphology of the egg, the reticulations on its surface, the micropylar region and micropylar structure, and the posterior filament coil were examined. In this study, the ultrastructure of L. tetraphylla egg was examined for the first time using scanning electron microscopy (SEM). As a result of the examination, it was detected for the first time that the posterior filament coil, which is rarely seen in odonate eggs, is also present in L. tetraphylla eggs. By comparing the L. tetraphylla egg with the eggs of I. ferox, I. rapax, I. australis, and L. africanus species, which are similar in terms of the posterior filament coil, the features that distinguish the L. tetraphylla egg.

Phage probe on RAFT polymer surface for rapid enumeration of E. coli K12.

This study presents a simple, fast, and sensitive label-free sensing assay for the precise enumeration of modeled pathogenic Escherichia coli K12 (E. coli K12) bacteria for the first time. The method employs the covalent binding bacteriophage technique on the surface of a reversible addition-fragmentation chain transfer (RAFT) polymer film. The Nyquist plots obtained from electrochemical impedance spectroscopy (EIS) identified the charge transfer resistance Rct was calculated from a suitable electrochemical circuit model through an evaluation of the relevant parameter after the immobilization of the bacteriophage and the binding of specific E. coli K12. The impedimetric biosensor reveals specific and reproducible detection with sensitivity in the linear working range of 104.2-107.0 CFU/mL, a limit of detection (LOD) of 101.3 CFU/mL, and a short response time of 15 min. The SERS response validates the surface roughness and interaction of the SERS-tag with E. coli K12-modified electrodes. Furthermore, the covalently immobilized active phage selectivity was proved against various non-targeting bacterial strains in the presence of targeted E.coli K12 with a result of 94 % specificity and 98 % sensitivity. Therefore, the developed phage-based electrode surface can be used as a disposable, label-free impedimetric biosensor for rapid and real-time monitoring of serum samples.

Gold-Coated Iron Composite Nanospheres Targeted the Detection of Escherichia coli.

We report the preparation and characterization of spherical core-shell structured Fe3O4-Au magnetic nanoparticles, modified with two component self-assembled monolayers (SAMs) consisting of 3-mercaptophenylboronic acid (3-MBA) and 1-decanethiol (1-DT). The rapid and room temperature synthesis of magnetic nanoparticles was achieved using the hydroxylamine reduction of HAuCl4 on the surface of ethylenediaminetetraacetic acid (EDTA)-immobilized iron (magnetite Fe3O4) nanoparticles in the presence of an aqueous solution of hexadecyltrimetylammonium bromide (CTAB) as a dispersant. The reduction of gold on the surface of Fe3O4 nanoparticles exhibits a uniform, highly stable, and narrow particle size distribution of Fe3O4-Au nanoparticles with an average diameter of 9 ± 2 nm. The saturation magnetization value for the resulting nanoparticles was found to be 15 emu/g at 298 K. Subsequent surface modification with SAMs against glucoside moieties on the surface of bacteria provided effective magnetic separation. Comparison of the bacteria capturing efficiency, by means of different molecular recognition agents 3-MBA, 1-DT and the mixed monolayer of 3-MBA and 1-DT was presented. The best capturing efficiency of E. coli was achieved with the mixed monolayer of 3-MBA and 1-DT-modified nanoparticles. Molecular specificity and selectivity were also demonstrated by comparing the surface-enhanced Raman scattering (SERS) spectrum of E. coli-nanoparticle conjugates with bacterial growth media.

The efficacy of 8% Arginine-CaCO3 applications on dentine hypersensitivity following periodontal therapy: a clinical and scanning electron microscopic study.

OBJECTIVES: Periodontal therapy is one of the etiological factors of dentine hypersensitivity (DH). This study aimed to evaluate the efficacy of %8Arginine-CaCO3 on DH that affects patients after periodontal treatment. STUDY DESIGN: Seventy-one teeth from the volunteers (n=36) with history of DH caused by periodontal therapy were included in this study, and randomly divided into two groups: group-1, who received 8%Arginine-CaCO3 and group-2, who received 1.23%NaF-gel. The clinical indices were recorded at first visit.DH was evaluated by using tactile, air-blast, and thermal stimuli. The subject's response was recorded at baseline, immediately (Day-0) and one month after the application. RESULTS AND CONCLUSIONS: The results were statistically analyzed, and it was found that 8% Arginine-CaCO3 treatment was more effective than 1.23% NaF-gel at time intervals. Sensitivity score differences between the groups were statistically significant at Day-28. The 8% Arginine-CaCO3 group exhibited statistically significant reduction in DH on three stimuli at baseline to Day-28. It was concluded that 8% Arginine-CaCO3 is more effective than 1.23% NaF-gel in reduction of patients' pain.

A new bio-active asymmetric-Schiff base: synthesis and evaluation of calf thymus DNA interaction, topoisomerase IIα inhibition, in vitro antiproliferative activity, SEM analysis and molecular docking studies.

In this paper, the asymmetric-Schiff base 2-(4-(2-hydroxybenzylideneamino)benzylideneamino)benzoic acid (SB-2) was newly synthesized and characterized by various spectroscopic methods. The interaction of SB-2 with calf thymus DNA was investigated by UV-vis, fluorescence spectroscopy and molecular docking methods. It was determined that SB-2 effectively binds to DNA via the intercalation mode. DNA electrophoretic mobility experiments displayed that topoisomerase IIα could not cleave pBR322 plasmid DNA in the presence of SB-2, confirming that the Schiff base acts as a topo II suppressor. In the molecular docking studies, SB-2 was found to show an affinity for both the DNA-topoisomerase IIα complex and the DNA. In vitro antiproliferative activity of SB-2 was screened against HT-29 (colorectal) and HeLa (cervical) human tumor cell lines by MTT assay. SB-2 diminished the cell viability in a concentration- and incubation time-dependent manner. The ability of SB-2 to measure DNA damage in tumor cells was evaluated with cytokinesis-block micronucleus assay after incubation 24 h and 48 h. Light and scanning electron microscopy experiments of tumor cells demonstrated an incubation time-dependent increase in the proportion of apoptotic cells (nuclear condensation and apoptotic bodies) suggesting that autophagy and apoptosis play a role in the death of cells. Based on the obtained results, it may be considered that SB-2 is a candidate for DNA-targeting antitumor drug.Communicated by Ramaswamy H. Sarma.

In vitro genotoxic and antigenotoxic effects of an exopolysaccharide isolated from Lactobacillus salivarius KC27L.

Exopolysaccharide isolated from Lactobacillus salivarius (new genus name Ligilactobacillus) KC27L strain (EPSKC27L) exhibits antioxidant properties with 1,1-diphenyl-2-picrylhydrazase (DPPH) radical and superoxide anion radical (O2-.) scavenging effect and iron ion (Fe2+) chelating activity. This study aimed to investigate the in vitro genotoxic effects of EPSKC27L alone (12.50, 25.00, 50.00, and 100.00 µg/mL) and its antigenotoxic activity against DNA damage induced by mitomycin-C (MMC; 0.20 µg/mL), methyl methanesulfonate (MMS; 5.00 µg/mL), and hydrogen peroxide (H2O2; 100 µM). For this purpose, chromosome aberration (CA), sister chromatid exchange (SCE), micronucleus (MN), and comet assays were performed in human peripheral lymphocytes. In addition, the structure of EPSKC27L was investigated in the scanning electron microscope (SEM). EPSKC27L alone did not cause a significant genotoxic effect in CA, SCE, MN, and comet tests. EPSKC27L significantly decreased the frequency of CA, SCE, and MN induced by MMC and MMS. EPSKC27L also significantly reduced DNA damage induced by H2O2. This study showed that the EPSKC27L alone has no genotoxic risk at these concentrations and shows antigenotoxic activity against MMC, MMS, and H2O2. Consequently, EPSKC27L was found to exhibit chemopreventive activity against genotoxic agents. This effect is believed to be due to the antioxidant properties of EPSKC27L.

Histopathologic and Spectrometric Evaluation of Bony Components of Synostotic Suture and Parietal Bone in Children with Sagittal Synostosis.

AIM: To understand the characterization of the ossification process both in the synostotic suture, and the adjacent parietal bone. MATERIAL AND METHODS: The surgical procedure for the 28 patients diagnosed with sagittal synostosis consisted of removing the synostotic bone as a whole, if possible, "Barrel-Stave" relaxation osteotomies, and strip osteotomies to the parietal and temporal bones perpendicular to the synostotic suture. The synostotic (group I) and parietal (group II) bone segments are obtained during osteotomies. Atomic absorption spectrometry was used to determine the amount of calcium in both groups, which is an indicator of ossification. Scanning electron microscopy and immunohistochemistry were employed to assess trabecular bone formation, osteoblastic density, and osteopontin, which is one of the in vivo indicators of new bone formation. RESULTS: Histopathologically, trabecular bone formation scores did not indicate any significant difference between the groups. However, the osteoblastic density and calcium accumulation in group I were higher than those in group II, and the difference was significant. Osteopontin staining scores in cells showing membranous and cytoplasmic staining with osteopontin antibodies significantly increased in group II. CONCLUSION: In this study, we found reduced differentiation of osteoblasts despite their increase in number. Moreover, the osteoblastic maturation rate was low in synostotic sutures, bone resorption becomes slower than new bone formation, and the remodeling rate is low in sagittal synostosis.

Morphology of dry-resistant eggs in parthenogenetic Heterocypris incongruens (Ramdohr, 1808) (Ostracoda, Crustacea).

It has been known that many organisms evolved to survive in temporary or ephemeral inland waters. Many of them have dry-resistant eggs against desiccation. The structural feature of egg shell is important because only this will ensure to survive the dry period. Structural features of egg shell in the parthenogenetic Heterocypris incongruens (Ramdohr, 1808) was investigated by scanning electron microscope. Results showed that egg shell structure consists of two distinct layers; an outer layer with holes or alveoli and an inner layer consisting of two dense sublayers. Also, structural similarities in egg-shell of H. incongruens and some other crustaceans which combat desiccation problem will be discussed.

Ultrastructure of aedeagus, spermatheca and ovipositor of Julodis ehrenbergii Laporte (Coleoptera: Buprestidae: Julodinae) by scanning electron microscope.

The paper presents unknown ultrastructure observed by scanning electron microscope (SEM) of aedeagus, spermatheca and ovipositor of Julodis ehrenbergii Laporte (Coleoptera: Buprestidae: Julodinae) from Turkey. The studied specimens were collected from Çankiri and Çorum provinces which are new provincial records for Turkey, in July and August 2021. The genus Julodis Eschscholtz includes 49 species in the Palearctic region, while it is represented by seven species in Turkey. One of them, J. ehrenbergii Laporte is represented by only the nominate subspecies in Turkey. As known, aedeagus, spermatheca and ovipositor are taxonomically important structures. Before the present study, however, there are no work on these structures of J. ehrenbergii Laporte. For this reason, ultrastructural and detailed investigations of aedeagus, spermatheca and ovipositor of J. ehrenbergii Laporte from Turkey were firstly studied with SEM and stereo microscope to obtain new diagnostic characters in the genus Julodis Eschscholtz. The female spermatheca is found insignificant and not diagnostic but aedeagus is found important for diagnosis. In addition, the ultrastructural characters of the ovipositor can also be useful for diagnosis. Photos in SEM as well as photos in the stereo microscope are also given in the text. HIGHLIGHTS: Ultrastructural and detailed investigations of aedeagus, spermatheca and ovipositor of Julodis ehrenbergii Laporte from Turkey were firstly studied with scanning electron microscope (SEM) and stereo microscope to obtain new diagnostic characters in the genus Julodis Eschscholtz. New diagnostic characters that can be distinguished by SEM have been revealed in aedeagus, spermatheca and ovipositor structures, which are used for morphological differentiation of species.

Construction of a sensitive and selective plasmonic biosensor for prostate specific antigen by combining magnetic molecularly-imprinted polymer and surface-enhanced Raman spectroscopy.

Selective and sensitive detection of cancer biomarkers in serum samples is critical for early diagnosis of cancer. Prostate specific antigen is an important biomarker of prostate cancer, which ranks high among cancer-related deaths of men over 50 years old. Herein, a novel analytical method was introduced for detection of PSA by combining high selectivity of molecularly-imprinted polymers and high sensitivity of surface-enhanced Raman spectroscopy (SERS). Firstly, magnetic nanoparticles were grafted with an imprinted layer by using tannic acid as a functional monomer, diethylenetriamine as a cross-linker and prostate specific antigen as a template molecule. Detailed surface characterization and re-binding experiment results indicated that the imprinting of the antigen was successful with an imprinting factor of 5.58. The prepared magnetic molecularly imprinted polymers (MMIPs) were used as an antibody-free capture probe and labeled with gold nanoparticles that were modified with anti-PSA and a Raman reporter, namely 5,5'-dithiobis-(2-nitrobenzoic acid). Thus, a plasmonic structure (sandwich complex) was formed between MMIP and the SERS label. The limit of detection and limit of quantification of the designed sensor were 0.9 pg/mL and 3.2 pg/mL, respectively. The sensor also showed high recovery rates (98.0-100.1% for healthy person and 99.0-101.3% for patient) with low standard deviations (less than 4.3% for healthy person and less than 3.3% for patient) for PSA in serum samples. Compared with the traditional immunoassays, the proposed method has several advantages like low cost, reduced detection procedure, fast response, high sensitivity and selectivity. It is believed that the proposed method can be potentially used for selective and sensitive determination of tumor marker of prostate cancer in clinical applications.

Escherichia coli Enumeration in a Capillary-Driven Microfluidic Chip with SERS.

Pathogen detection is still a challenging issue for public health, especially in food products. A selective preconcentration step is also necessary if the target pathogen concentration is very low or if the sample volume is limited in the analysis. Plate counting (24-48 h) methods should be replaced by novel biosensor systems as an alternative reliable pathogen detection technique. The usage of a capillary-driven microfluidic chip is an alternative method for pathogen detection, with the combination of surface-enhanced Raman scattering (SERS) measurements. Here, we constructed microchambers with capillary microchannels to provide nanoparticle-pathogen transportation from one chamber to the other. Escherichia coli (E. coli) was selected as a model pathogen and specific antibody-modified magnetic nanoparticles (MNPs) as a capture probe in a complex milk matrix. MNPs that captured E. coli were transferred in a capillary-driven microfluidic chip consisting of four chambers, and 4-aminothiophenol (4-ATP)-labelled gold nanorods (Au NRs) were used as the Raman probe in the capillary-driven microfluidic chip. The MNPs provided immunomagnetic (IMS) separation and preconcentration of analytes from the sample matrix and then, 4-ATP-labelled Au NRs provided an SERS response by forming sandwich immunoassay structures in the last chamber of the capillary-driven microfluidic chip. The developed SERS-based method could detect 101-107 cfu/mL of E. coli with the total analysis time of less than 60 min. Selectivity of the developed method was also tested by using Salmonella enteritidis (S. enteritidis) and Staphylococcus aureus (S. aureus) as analytes, and very weak signals were observed.