Expedient liquid chromatographic method with fluorescence detection for montelukast sodium in micro-samples of plasma

This study describes an expedient assay for the analysis of the asthma medication, montelukast sodium (Singulair, MK-0476), in human plasma samples. After a simple extraction of the plasma, the drug and internal standard, quinine bisulfate, were measured by HPLC. The chromatographic system consisted of a single pump, a refrigerated autosampler, a C₈ 4-μm particle size radial compression cartridge at 40°C and a fluorescence detector with the excitation and emission wavelengths set at 350 and 400 nm, respectively. The mobile phase which was delivered at 1.0 ml/min, was prepared by adding 200 ml of 0.025 M sodium acetate, pH adjusted to 4.0 with acetic acid, to 800 ml of acetonitrile, with 50 μl triethylamine. With a run time of only 10 min per sample, this assay had an overall recovery of >97% with a detection limit of 1 ng/ml. The inter- and intra-run relative standard deviations at 0.05, 0.2 and 1.0 μg/ml were all <9.2%, while the analytical recovery at the same concentrations were within 7.7% of the amount added.     

Changing seasonality and phenological responses of free-living male arctic ground squirrels: the importance of sex

Many studies have addressed the effects of climate change on species as a whole; however, few have examined the possibility of sex-specific differences. To understand better the impact that changing patterns of snow-cover have on an important resident Arctic mammal, we investigated the long-term (13 years) phenology of hibernating male arctic ground squirrels living at two nearby sites in northern Alaska that experience significantly different snow-cover regimes. Previously, we demonstrated that snow-cover influences the timing of phenological events in females. Our results here suggest that the end of heterothermy in males is influenced by soil temperature and an endogenous circannual clock, but timing of male emergence from hibernation is influenced by the timing of female emergence. Males at both sites, Atigun and Toolik, end heterothermy on the same date in spring, but remain in their burrows while undergoing reproductive maturation. However, at Atigun, where snowmelt and female emergence occur relatively early, males emerge 8 days earlier than those at Toolik, maintaining a 12-day period between male and female emergence found at each site, but reducing the pre-emergence euthermic period that is critical for reproductive maturation. This sensitivity in timing of male emergence to female emergence will need to be matched by phase shifts in the circannual clock and responsiveness to environmental factors that time the end of heterothermy, if synchrony in reproductive readiness between the sexes is to be preserved in a rapidly changing climate.     

Seasonality,habitat type and locality influenced bird assemblage structure in Nigeria

This is the first report of the avian assemblage in the study area of Dutse, Nigeria. In addition to recording bird species, the effects of season, dominant vegetation structure, locality and anthropogenic activities on bird abundance, species richness and diversity were investigated. Using the point transect method, 264 points on 48 km of transect were used to count birds between 06:30 and 11:00 from August 2015 to February 2016. A total of 122 bird species of 41 families were recorded. Highest bird species richness was recorded in Warwade, highest abundance in Model, and highest diversity in Malamawa. The dry season and woodland habitat showed higher bird species richness, abundance and diversity than the wet season and shrubland habitat. Tree density was more important in increasing bird abundance than shrub density. Small-scale anthropogenic activities and habitat modification, such as farming, grazing, wood removal and human interference did not appear to have impacted the birds; however, loss of high tree-density woodland habitats may pose a major threat to the bird community in Dutse. The presence of birds of concern in the area suggests the need for conservation efforts of avifauna and as well as the forested habitats in Dutse.     

Quantifying the ultrastructure changes of air-dried and irradiated human amniotic membrane using atomic force microscopy: a preliminary study

Air-dried and sterilized amnion has been widely used as a dressing to treat burn and partial thickness wounds. Sterilisation at the standard dose of 25 kGy was reported to cause changes in the morphological structure as observed under the scanning electron microscope. This study aimed to quantify the changes in the ultrastructure of the air-dried amnion after gamma-irradiated at several doses by using atomic force microscope. Human placentae were retrieved from mothers who had undergone cesarean elective surgery. Amnion separated from chorion was processed and air-dried for 16 h. It was cut into 10?×?10 mm, individually packed and exposed to gamma irradiation at 5, 15, 25 and 35 kGy. Changes in the ultrastructural images of the amnion were quantified in term of diameter of the epithelial cells, size of the intercellular gap and membrane surface roughness. The longest diameter of the amnion cells reduced significantly after radiation (p?<?0.01) however the effect was not dose dependent. No significant changes in the shortest diameter after radiation, except at 35 kGy which decreased significantly when compared to 5 kGy (p?<?0.01). The size of the irradiated air-dried amnion cells reduced in the same direction without affecting the gross ultrastructure. At 15 kGy the intercellular gap decreased significantly (p?<?0.01) with Ra and Rq, values reflecting surface roughness, were significantly the highest (p?<?0.01). Changes in the ultrastructure quantified by using atomic force microscope could complement results from other microscopic techniques.     

Iraqi propolis increases degradation of IL-1β and NLRC4 by autophagy following Pseudomonas aeruginosa infection

Autophagy is a cellular process for maintaining cellular homeostasis. This process can be induced by different factors, such as immune stimuli and pathogen-associated molecules. Autophagy has an important role in the control of IL-1β secretion by macrophages and other cell types. In present study, we describe a novel role for Iraqi propolis affecting autophagy in controlling the secretion of IL-1β in bone-marrow macrophages (BMDMs). After infection with Pseudomonas aeruginosa in the presence of propolis, the degradation of IL-1β was induced, and the activity of inflammasome was reduced. Iraqi propolis-induced autophagy in in vitro and in vivo models decreased the levels of IL-1β and caspase-1. Results indicated that IL-1β pathway production is regulated by autophagy via two different novel mechanisms, namely, regulation of the activation of NLRC4 inflammasome and IL-1β targeting for lysosomal degradation.     

Biogenic synthesis of copper oxide nanoparticles using olea europaea leaf extract and evaluation of their toxicity activities: An in vivo and in vitro study

Copper oxide nanoparticles (CUNPs) were synthesized using Olea europaea leaf extract as reducing and protecting agent. The formation of nanoparticles was observed through a color change from yellowish to brownish black. The CUNPs were confirmed with UV–Vis spectrophotometer, which revealed a peak absorbance at 289 nm. The synthesized CUNPs were characterized by XRD, FTIR, SEM, and TEM. The XRD pattern revealed that CUNPs were crystalline in nature with a diameter around 20 nm. FTIR spectral analysis showed that CUNPs were capped with plant constituents. From SEM and TEM analyses, the CUNPs were generally found to be spherical in shape, and the size range was 20–50 nm. Free radical scavenging potential of CUNPs against DPPH was confirmed by its stable antioxidant effects. In addition, the toxicity of CUNPs in mice was also assessed by body weight and weights of liver, kidneys, spleen, and thymus. The immune response in mice was signaled through an obvious change in spleen and thymus index, with a decrease of ADA enzyme activity in serum, spleen, and thymus after CUNPs treatment. The CUNPs were found to exert cell growth arrest against AMJ‐13 and SKOV‐3 cancer cells in a dose‐dependent manner and induce cell death by apoptosis. Less significant cytotoxic effect was observed in normal dermal fibroblast cells. These findings suggest that CUNPs may have the potential to be anticancer agents. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:218–230, 2018     

Evaluation of Cryptosporidium parvum Genotyping Techniques

We evaluated the specificity and sensitivity of 11 previously described species differentiation and genotyping PCR protocols for detection of Cryptosporidium parasites. Genomic DNA from three species of Cryptosporidium parasites (genotype 1 and genotype 2 of C. parvum, C. muris, and C. serpentis), two Eimeria species (E. neischulzi and E. papillata), and Giardia duodenalis were used to evaluate the specificity of primers. Furthermore, the sensitivity of the genotyping primers was tested by using genomic DNA isolated from known numbers of oocysts obtained from a genotype 2 C. parvum isolate. PCR amplification was repeated at least three times with all of the primer pairs. Of the 11 protocols studied, 10 amplified C. parvum genotypes 1 and 2, and the expected fragment sizes were obtained. Our results indicate that two species-differentiating protocols are not Cryptosporidium specific, as the primers used in these protocols also amplified the DNA of Eimeria species. The sensitivity studies revealed that two nested PCR-restriction fragment length polymorphism (RFLP) protocols based on the small-subunit rRNA and dihydrofolate reductase genes are more sensitive than single-round PCR or PCR-RFLP protocols.     

Identification of a critical motif responsible for gating of Kir2.3 channel by intracellular protons.

Protons are involved in gating Kir2.3. To identify the molecular motif in the Kir2.3 channel protein that is responsible for this process, experiments were performed using wild-type and mutated Kir2. 3 and Kir2.1. CO2 and low pHi strongly inhibited wild-type Kir2.3 but not Kir2.1 in whole cell voltage clamp and excised inside-out patches. This CO2/pH sensitivity was completely eliminated in a mutant Kir2.3 in which the N terminus was substituted with that in Kir2.1, whereas a similar replacement of its C terminus had no effect. Site-specific mutations of all titratable residues in the N terminus, however, did not change the CO2/pH sensitivity. Using several chimeras generated systematically in the N terminus, a 10-residue motif near the M1 region was identified in which only three amino acids are different between Kir2.3 and Kir2.1. Mutations of these residues, especially Thr53, dramatically reduced the pH sensitivity of Kir2.3. Introducing these residues or even a single threonine to the corresponding positions of Kir2.1 made the mutant channel pH-sensitive. Thus, a critical motif responsible for gating Kir2.3 by protons was identified in the N terminus, which contained about 10 residues centered by Thr53.     

Macrocyclic inhibitors of Factor XIa: Discovery of alkyl-substituted macrocyclic amide linkers with improved potency

Optimization of macrocyclic inhibitors of FXIa is described which focused on modifications to both the macrocyclic linker and the P1 group. Increases in potency were discovered through interactions with a key hydrophobic region near the S1 prime pocket by substitution of the macrocyclic linker with small alkyl groups. Both the position of substitution and the absolute stereochemistry of the alkyl groups on the macrocyclic linker which led to improved potency varied depending on the ring size of the macrocycle. Replacement of the chlorophenyltetrazole cinnamide P1 in these optimized macrocycles reduced the polar surface area and improved the oral bioavailability for the series, albeit at the cost of a decrease in potency.