Antibiotic Supplements Affect Electrophysiological Properties and Excitability of Rat Hippocampal Pyramidal Neurons in Primary Culture

Introduction: Antibiotic supplements are regularly used in neuronal culture media to control contamination; however, they can interfere with the neuronal excitability and affect electrophysiological properties. Therefore, in this study, the effect of penicillin/streptomycin supplements on the spontaneous electrophysiological activity of hippocampal pyramidal neurons was examined. Methods: Electrophysiological whole-cell patch-clamp recordings from rat hippocampal pyramidal cells in primary culture were performed to investigate the effects of antibiotic supplements on the intrinsic excitability of cultured cells. Results: The present findings indicated that presence of antibiotic supplements (penicillin/streptomycin) in the culture medium altered the intrinsic electrical activity of hippocampal pyramidal neurons in primary culture. These alterations included: 1) depolarized resting membrane potential; 2) a significant enhancement in the after-hyperpolarization amplitude; 3) a significant increase in the area under the action potential and in the decay and rise time of the action potential; 4) a significant broadening of action potential and 5) a significant reduction in the firing frequency. Conclusion: These findings suggest that addition of antibiotic supplements to culture media influences the neuronal excitability and alters the electrophysiological properties of cultured neurons, possibly through changing the ionic conductance underlying neuronal excitability. Key Words: Primary cell culture, Patch-clamp techniques, Hippocampus     

Morphological, optical and electromagnetic characterization of polybutylene terephthalate/silica nanocomposites

Polybutylene terephthalate (PBT) composites containing 1 %, 3 % and 5 % silica nanoparticles were prepared by melt compounding method. The characteristics of the nanocomposites were investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), tensile strength test (TST), reflectance spectroscopy (RS), electromagnetic transition test (ETT) and atomic force microscopy (AFM). XRD method showed successful incorporation of silica into the polymer matrix. SEM and AFM results showed the presence of silica aggregates on the surface of PBT sheets. Thermal analysis results demonstrated some changes in crystallinity of PBT after addition of silica. The PBT/silica nanocomposites were found to have higher electromagnetic reflection compared with the pure PBT. Silica nanoparticle is thus suggested as a good candidate for electromagnetic shielding purposes.     

SiO2-kaolinite affecting the surface properties of ternary poly(vinyl chloride)/silica/kaolinite nanocomposites

The products from the dispersion of nanoscale particulates such as the layered clays or the spherical inorganic minerals within the polymeric matrices are called polymeric nanocomposites. In this paper, we prepared poly(vinyl chloride) (PVC) based nanocomposites containing SiO₂-kaolinite by melt compounding. The influence of SiO₂-kaolinite on the surface properties of PVC was investigated by the use of various surface analysis techniques including a ttenuated total reflectance spectroscopy (ATR), wide angle X-ray diffractometry (WAXD), atomic force microscopy (AFM), scanning electron microscopy (SEM), electron dispersive X-ray spectrometer (EDX), contact angle measurement (CAM), and reflectance spectroscopy (RS). ATR spectroscopy showed possible interaction between layered kaolinite and PVC at surface. Microscopic methods illustrated an increased surface roughness compared to the pure PVC. Contact angle measurements of the resultant PVC nanocomposites demonstrated that the wettability of substrates depends on the surface interactions between kaolinite layers and PVC matrix. Optical properties of nanocomposite films were finally measured by the aid of reflectance spectrophotometer. It can be seen from optical studies that reflectance values were increased after incorporation of SiO₂-kaolinite in nanocomposite.     

Rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) anesthesia with myrcene: efficacy and physiological responses in comparison with eugenol

The aim of the present study was to investigate anesthetic efficacy of myrcene in rainbow trout (Oncorhynchus mykiss) along with the fish biochemical response to anesthesia in comparison with eugenol. In the first experiment, 240 fish were stocked in 12 tanks and acclimatized to experimental conditions for 2 weeks. Then, the fish of each tank were subjected to one concentration of either eugenol (12, 20, 30, 50, 80, and 130 μL/L) or myrcene (100, 150, 200, 300, 400, and 500 μL/L) concentrations. Induction time of and recovery time from anesthesia were recorded for each fish separately. Using these results, desired concentrations to induce anesthesia within 60, 180, 300, and 600 s were determined, being 81, 30, 19, and 10 μL/L eugenol and 531, 251, 177, and 111 μL/L myrcene. In the second experiment, 96 fish were stocked in 8 tanks. Six fish were netted from each tank and exposed to the calculated eugenol or myrcene concentrations. Blood samples were taken after the fish reached anesthesia. The results showed that there was no significant difference in serum lactate, alanine transaminase, and alkaline phosphatase. Increase in the induction time of anesthesia resulted in increased serum glucose with no significant difference between the anesthetics. Increase in induction time of anesthesia led to increase in serum lactate dehydrogenase activity in the eugenol-anesthetized fish and aspartate transaminase activity in myrcene-anesthetized fish. In conclusion, myrcene is capable to anesthetize rainbow trout, but at higher concentrations compared to eugenol. In addition, biochemical analysis showed that increase in induction time of anesthesia leads to hyperglycemia and increase in AST and LDH activities depending on anesthetic type.     

Expression of ROR1 in patients with renal cancer--a potential diagnostic marker

Background: The ectopic expression of receptor tyrosine kinase Ror1 has been reported in patients with hematological malignancies such as chronic lymphocytic leukemia and acute lymphoblastic leukemia. Here we report, for the first time, expression of ROR1 gene in both tumor tissues and peripheral blood mononuclear cells (PBMC) from patients with renal cancer (RC). Methods: In the current study, the expression of ROR1 gene was semi-quantitatively measured in PBMC and tumor tissues from 16 RC patients as well as PBMC from 22 healthy individuals relative to the expression of the housekeeping gene phosphoglucomutase 1 by RT-PCR. Results: Our results showed that ROR1 was expressed at gene level in 81.3% of renal tumor tissues (13 out of 16) whereas it was expressed in 94% of PBMC from RC patients (15 out of 16). A weak expression of ROR1 was observed in PBMC of 4 out of 22 healthy individual. A significant expression of ROR1 was observed in PBMC from RC patients when compared to that in PBMC from normal healthy individuals (P<0.001). The expression of ROR1 in PBMC may reflect a shedding of tumor cells into blood stream. Conclusion: We conclude that detection of a high level of ROR1 expression in blood cells might assist in early detection of renal malignancies, providing taking into consideration the clinical symptoms of the disease. Key Words: ROR1, Ectopic expression, Renal cancer     

A rapid diagnostic assay for detection and quantification of the causal agent of strawberry wilt from field samples

Verticillium dahliae Kleb, the cause of Verticillium wilt disease, is a destructive pathogen that leads to severe yield losses in strawberry fields and thus considerable economic damages. Although rapid identification and detection methods are becoming available more, pathogen quantification remains one of the main challenges in the disease management. In this study, a real-time polymerase chain reaction (rtPCR) assay was developed to quantitatively assess V. dahliae abundance directly from affected roots and soil collected from different areas in Estonia. A specific primer pair based on the ribosomal DNA (rDNA) internally transcribed spacer was designed for SYBR Green-based assay. Strawberry plant and soil samples were randomly collected from different areas in Estonia and analyzed for V. dahliae by soil plating technique and rtPCR assay. The assay was specific for V. dahliae so that the minimum detection limit was 0.93?pg?µl^?1 of pathogen DNA and the lowest amount of V. dahliae detected in soil was 10.48?pg?µl^?1 of target DNA corresponding to one microsclerotia per gram of soil. This technique allowed rapid detection and quantification of the pathogen DNA at the picogram level in soils and even in symptomless plants, facilitating the screening of the pathogen in diverse areas. This is the first study about the rtPCR technique being used successfully to assess populations of V. dahliae with high specificity and sensitivity in Estonia strawberry fields. Results of this research can be useful for growers and agricultural organizations to improve available disease management strategies against Verticillium wilt.     

Functional involvement of Ca(2+) and Ca(2+)-activated K(+) channels in anethol-induced changes in Ca(2+) dependent excitability of F1 neurons in Helix aspersa

The effects of anethol, the major component of anise oil, on the Ca²⁺-dependent excitability and afterhyperpolarization (AHP) in snail neurons were examined using intracellular recording. Anethol (0.5%) significantly broadened the spike, reduced the firing frequency and enhanced the AHP amplitude. In contrast, anethol (2%) significantly increased the firing frequency and decreased the AHP. Blockade of Ca²⁺ channels after anethol application depolarized the membrane potential and significantly reduced the firing rate. Furthermore, in the presence of anethol (0.5%) a significant decrease in the AHP was observed by Ca²⁺ channels blockage. Here, anethol-induced functional modification of Ca²⁺ and Ca²⁺-activated K⁺ channels is suggested.     

Description of a new species of seed-gall nematode, <Emphasis Type="Italic">Anguina obesa</Emphasis> n. sp. (Nematoda: Anguinidae) from northern Iran,and its phylogenetic relations with other species and genera

Anguina obesa n. sp., a new species of the genus, causing small seed galls inside the ovaries of foxtail weed plants (Alopecurus mysuroides Huds.) is described and illustrated based on its morphological and molecular characters. The new species is characterized by its 1516–2564 μm long obese females irregularly ventrally curved after fixation, having six lines in lateral fields, 6–9 μm long stylet with well-developed rounded knobs, constriction at junction of isthmus with the pharyngeal bulb, monodelphic-prodelphic female reproductive system, and conical, 60–80 μm long tail. Males of the new species are characterized with their slender 936–1420 μm long body, 25–30 μm long tylenchoid spicules, and bursa not reaching tail tip. Second stage juveniles of the new species were also common inside the galls and also recovered from soil in type locality. The new species is morphologically close to Anguina agropyronifloris, A. amsinckiae, A. paludicola and A. tumefaciens, but is more closely related to A. paludicola, from which it can be separated based on differences in morphological characters and internal transcribed spacer sequence. In Bayesian inference using sequences of the aforementioned genomic fragment, the new species formed a clade with A. agrostis, A. funesta, A. graminis, A. phalaridis and some unidentified isolates, with robust Bayesian posterior probability (BPP). The morphologically closest species, A. paludicola, occupied a separate position, outside of the clade containing the new species. The sequences of two other genomic fragments, 18S and 28S rDNA (D2/D3 region) were also made available for the new species. Morphological comparisons of the new species with the related species are discussed.