Simultaneous quantification of free nucleotides in complex biological samples using ion pair reversed phase liquid chromatography isotope dilution tandem mass spectrometry

A new sensitive and accurate analytical method has been developed for quantification of intracellular nucleotides in complex biological samples from cultured cells of different microorganisms such as Saccharomyces cerevisiae, Escherichia coli, and Penicillium chrysogenum. This method is based on ion pair reversed phase liquid chromatography electrospray ionization isotope dilution tandem mass spectrometry (IP-LC-ESI-ID-MS/MS. A good separation and low detection limits were observed for these compounds using dibutylamine as volatile ion pair reagent in the mobile phase of the LC. Uniformly ¹³C-labeled isotopes of nucleotides were used as internal standards for both extraction and quantification of intracellular nucleotides. The method was validated by determining the linearity, sensitivity, and repeatability.     

Spatial distribution and diversity of commercial demersal fish species in the shelf waters of the Caspian Sea

The Caspian Sea (CS) is a unique ecosystem known for its several fish species, especially sturgeons. To exhaustively manage the fish stocks of this ecosystem, detailed knowledge of species composition, abundance, distribution, and the habitat traits of the living organisms is necessary. This study analyzed the diversity, spatial, and seasonal distribution of commercial demersal fish species, and examined the relationships between community structure and environmental variability in Iranian shelf waters of the CS. For this purpose, seasonally fish sampling took place between 2009 and 2011 with a bottom trawl. Among 11 fish species captured, Chelon spp., Rutilus kutum, and Vimba vimba showed the highest abundances (i.e. 88.49%, 10.67%, and 0.69%, respectively). Univariate and multivariate analyses showed differences in commercial fish assemblages (abundance and species richness) according to seasons and regions. Distance-based Linear Model (DisTLM) showed that eight environmental variables display significant linear relationships with the fauna resemblance matrix (p < .05). Based on AIC criteria, the combination of silt&clay, TOM, longitude, depth, bottom, and surface temperature used to build the parsimonious DisTLM model explain 67.03% of the total variability. The results revealed a biogeographical and temporal gradient from the west to the east and summer to winter, in terms of commercial demersal fish assemblages and species diversity, as a consequence of different geomorphological, bottom substratum conditions and benthic communities.     

Fabrication of an Electrochemical Sensor Based on Gold Nanoparticles/Carbon Nanotubes as Nanocomposite Materials: Determination of Myricetin in Some Drinks

In this paper, the electrochemical behavior of myricetin on a gold nanoparticle/ethylenediamine/multi-walled carbon-nanotube modified glassy carbon electrode (AuNPs/en/MWCNTs/GCE) has been investigated. Myricetin effectively accumulated on the AuNPs/en/MWCNTs/GCE and caused a pair of irreversible redox peaks at around 0.408 V and 0.191 V (vs. Ag/AgCl) in 0.1 mol L⁻¹ phosphate buffer solution (pH 3.5) for oxidation and reduction reactions respectively. The heights of the redox peaks were significantly higher on AuNPs/en/MWNTs/GCE compare with MWCNTs/GC and there was no peak on bare GC. The electron-transfer reaction for myricetin on the surface of electrochemical sensor was controlled by adsorption. Some parameters including pH, accumulation potential, accumulation time and scan rate have been optimized. Under the optimum conditions, anodic peak current was proportional to myricetin concentration in the dynamic range of 5.0×10⁻⁸ to 4.0×10⁻⁵ mol L⁻¹ with the detection limit of 1.2×10⁻⁸ mol L⁻¹. The proposed method was successfully used for the determination of myricetin content in tea and fruit juices.     

The Effect of a Yeast Probiotic on Acute Diarrhea in Children

A probiotic is a living micro-organism administered to promote the health of the host by treating or preventing infections owing to strains of pathogens. Saccharomyces boulardii is a nonpathogen yeast that has a direct inhibitory effect on the growth of many pathogens, an anti-secretory effect and a trophic effect on enterocytes. The aim of this study was to determine the effect of S. boulardii on diarrhea in children. The children from 6 months to 6 years of age with acute watery diarrhea admitted in pediatric clinic in Kashan in 2012 were included in this trial. Exclusion criteria were high fever (T > 38.5 °C), severe dehydration, bloody diarrhea, severe malnutrition, using of antibiotics, anti-diarrheal or antifungal drugs and children with more than one complain. Two hundred patients were assigned into two groups: A total of 100 patients were treated with S. boulardii in addition to ORS (case group) and 100 patients were given placebo in addition to ORS (control group). The duration of diarrhea and frequency of stools were recorded by asking the mothers of the children every day. The results showed that the defecation frequency after second day of treatment in the case group was significantly less than the control group (P = 0.001) and the mean numbers of days of diarrhea was significantly lower in the case group (P = 0.001). The result of this study confirms that S. boulardii reduces the frequency of stool and duration of illness in children.     

Determination of the Shear Force at the Balance between Bacterial Attachment and Detachment in Weak-Adherence Systems, Using a Flow Displacement Chamber

We introduce a procedure for determining shear forces at the balance between attachment and detachment of bacteria under flow. This procedure can be applied to derive adhesion forces in weak-adherence systems, such as polymer brush coatings, which are currently at the center of attention for their control of bacterial adhesion and biofilm formation.     

Simultaneous and sensitive determination of a quaternary mixture of AA, DA, UA and Trp using a modified GCE by iron ion-doped natrolite zeolite-multiwall carbon nanotube

The evaluation of a novel modified glassy carbon electrode modified with iron ion-doped natrolite zeolite-multiwalled carbon nanotube for the simultaneous and sensitive determination of ascorbic acid (AA), dopamine (DA), uric acid (UA) and tryptophan (Trp) has been described. The measurements were carried out using cyclic voltammetry in buffer solution with pH 1. This modified electrode exhibits potent and persistent electroxidation behavior followed by well-separated oxidation peaks towards AA, DA, UA and Trp with increasing of the oxidation current. For the quaternary mixture containing AA, DA, UA and Trp, the 4 compounds can well separate from each other at the scan rate of 100 mVs(-1) with a potential difference of 270 mV, 150 mV and 260 mV for the oxidation peak potentials of AA-DA, DA-UA and UA-Trp, respectively, which was large enough to simultaneous determine AA, DA, UA and Trp. The catalytic peak current obtained, was linearly dependent on the AA, DA, UA and Trp concentrations in the range of 7.77-833 μM, 7.35-833 μM, 0.23-83.3 μM and 0.074-34.5 μM and the detection limits for AA, DA, UA and Trp were 1.11, 1.05, 0.033 and 0.011 μM, respectively. The analytical performance of this sensor has been evaluated for simultaneous detection of AA, DA, UA and Trp in human serum and urine samples.     

Distribution and possible function of an adrenomedullin-like peptide in the developing chick limb bud

Adrenomedullin (AM) is a multifunctional peptide that exhibits discrete domains of expression during mouse embryogenesis consistent with a role in regulating growth and differentiation during morphogenesis. Here we report that AM immunoreactivity is present at high levels throughout the apical ectodermal ridge (AER) of the chick limb bud as the AER is directing the outgrowth and patterning of underlying limb mesoderm. Immunostaining is particularly strong along the surfaces of the contiguous cells of the AER. AM immunoreactivity attenuates as the AER regresses and is absent from the distal apical ectoderm of stage 20 limbless mutant limb buds which fail to develop an AER. To explore the possible role of AM in AER activity, we examined the effect of exogenous AM and an AM inhibitor on the in vitro morphogenesis of limb mesoderm, cultured in the presence and absence of the AER. Although exogenous AM cannot substitute for the AER in promoting outgrowth of limb mesoderm in vitro, a specific AM antagonist, AM(22-52), impairs the outgrowth and proliferation of limb mesoderm cultured in the presence of the AER. This is consistent with the possibility that inhibition of endogenous AM activity in the AER impairs the ability of the AER to promote limb morphogenesis. Taken together, these studies suggest that an AM-like molecule may function in an autocrine fashion to regulate some aspect of AER activity.     

Screening yeast cells for absorption of selenium oxyanions

Certain yeast cells on solid nutrient medium produced colonies surrounded by a light zone of selenite absorption. This screening procedure resulted in the selection of 22 strains out of 200 isolates with different Se⁴⁺-absorbing capacity ranging from 16 to 98.8 g Se⁴⁺ g^–1 l^–1 h^–1. The highest rate of Se⁴⁺ elimination from the Na₂SeO₃ solution was observed with an oval shaped, cream pigmented fermentative yeast, tentatively called Candida sp. strain MS4. This strain was isolated from wastewater and found to accumulate selenium oxyanions. Se⁴⁺ uptake involved both inactive and active phenomena. The amounts of selenium (initial concentration 2 mg Se⁴⁺ l^–1) removed from aqueous solution by inactive and active phenomena were 667 g Se⁴⁺ g^–1 l^–1, and 1580 g Se⁴⁺ g^–1 l^–1, respectively. The strain also removed selenate inactively (135 g Se⁶⁺ g^–1 l^–1).