Mathematical modelling of human P2X-mediated plasma membrane electrophysiology and calcium dynamics in microglia

Regulation of cytosolic calcium (Ca²⁺) dynamics is fundamental to microglial function. Temporal and spatial Ca²⁺ fluxes are induced from a complicated signal transduction pathway linked to brain ionic homeostasis. In this paper, we develop a novel biophysical model of Ca²⁺ and sodium (Na⁺) dynamics in human microglia and evaluate the contribution of purinergic receptors (P2XRs) to both intracellular Ca²⁺ and Na⁺ levels in response to agonist/ATP binding. This is the first comprehensive model that integrates P2XRs to predict intricate Ca²⁺ and Na⁺ transient responses in microglia. Specifically, a novel compact biophysical model is proposed for the capture of whole-cell patch-clamp currents associated with P2X₄ and P2X₇ receptors, which is composed of only four state variables. The entire model shows that intricate intracellular ion dynamics arise from the coupled interaction between P2X₄ and P2X₇ receptors, the Na⁺/Ca²⁺ exchanger (NCX), Ca²⁺ extrusion by the plasma membrane Ca²⁺ ATPase (PMCA), and Ca²⁺ and Na⁺ leak channels. Both P2XRs are modelled as two separate adenosine triphosphate (ATP) gated Ca²⁺ and Na⁺ conductance channels, where the stoichiometry is the removal of one Ca²⁺ for the hydrolysis of one ATP molecule. Two unique sets of model parameters were determined using an evolutionary algorithm to optimise fitting to experimental data for each of the receptors. This allows the proposed model to capture both human P2X₇ and P2X₄ data (hP2X₇ and hP2X₄). The model architecture enables a high degree of simplicity, accuracy and predictability of Ca²⁺ and Na⁺ dynamics thus providing quantitative insights into different behaviours of intracellular Na⁺ and Ca²⁺ which will guide future experimental research. Understanding the interactions between these receptors and other membrane-bound transporters provides a step forward in resolving the qualitative link between purinergic receptors and microglial physiology and their contribution to brain pathology.     

Gene delivery in adherent and suspension cells using the combined physical methods

Cytotechnology - Physical methods are widely utilized to deliver nucleic acids into cells such as electro-transfection or heat shock. An efficient gene electro-transfection requires the best...     

Persian sturgeon insulin-like growth factor I: molecular cloning and expression during various nutritional conditions

The effects of different periods of starvation (1, 2, 3, and 4 weeks) and subsequent re-feeding (over a 4 week) on the compensatory growth performance and insulin-like growth factor I (IGF-I) mRNA expression in liver and white muscle were investigated in juvenile Persian sturgeon (Acipenser persicus). First, a fragment of 617 nucleotides coding for IGF-I was cloned from liver, which included an open reading frame of 486 nucleotides, encoding a 162 amino acid preproIGF-I. This is composed of a 45 aa for signal peptide, a 117 aa for the mature peptide comprising the B, C, A, and D domains, and a 47 aa for E domain. The mature Persian sturgeon IGF-I exhibits high sequence identities with other sturgeon species and teleost, ranging between 68 and 95 %. The pattern of IGF-I mRNA expression in the liver and white muscle was measured in response to different periods of starvation and subsequent re-feeding. Nutritional status influenced IGF-I mRNA expression pattern in both liver and muscle. IGF-I mRNA expression in the liver increased during starvation, before decreasing after re-feeding. Furthermore, white muscle IGF-I mRNA expression showed better responses to nutritional status and decreased following starvation and increased by re-feeding. However, changes in the expression of IGF-I mRNA were not significantly different between any of the treatments in both tissues. These data suggest that muscle and liver IGF-I mRNA expression do not have a regulatory role for somatic growth induced by compensatory growth in Persain sturgeon.     

New data on the spider fauna of Iran (Arachnida: Araneae)

We present the results of some spider surveys that were conducted in different parts of Iran. Faunistic records for 30 species which are new to Iran are presented here. Biogeographic data are provided for each species. Two families, namely Liocranidae and Mimetidae, are recorded for the first time from the country.     

Integrated pathway-based and network-based analysis of GC-MS rice metabolomics data under diazinon stress to infer affected biological pathways

Diazinon insecticide is widely applied in rice (Oryza sativa L.) fields in Iran. However, concerns are now being raised about its potential adverse impacts on rice. In this study, a time-course metabolic change in rice plants was investigated after diazinon treatment using gas chromatography-mass spectrometry (GC-MS) and subsequently three different methods, MetaboAnalyst, MetaboNetwork, and analysis of reporter reactions, as a potential multivariate method were used to find the underlying changes in metabolism with stronger evidence in order to link differentially expressed metabolites to biological pathways. Results clearly showed the similarity of acetylcholinesterase (AChE) of rice plants to that of animals in terms of its inhibitability by diazinon and emphasized that subsequent accumulation of AChE mainly affects the metabolism of osmolites and tricarboxylic acid intermediates subsequent accumulation of ACh mainly affects the metabolism of osmolites and TCA intermediates.     

Genetic variation and evolutionary origins of parthenogenetic Artemia (Crustacea: Anostraca) with different ploidies

Using two nuclear (ITS1 and Na⁺/K⁺ ATPase) and three mitochondrial (COI, 16S and 12S) markers, we determined the genetic variation and evolutionary relationship of parthenogenetic and bisexual Artemia. Our analyses revealed that mitochondrial genes had higher genetic variation than nuclear genes and that the 16S showed more variety than the other mitochondrial genes in parthenogenetic populations. Triploid parthenogens showed lower genetic variation than diploid ones, whereas the tetra‐ and pentaploids had greater genetic distance than diploid parthenogens. No shared haplotype was found between individuals of parthenogenetic populations and Asian bisexual species with the exception of Na⁺/K⁺ ATPase (Artemia tibetiana). Only mitochondrial markers can demonstrate phylogenetic relationships, and showed that the parthenogenetic Artemia is a polyphyletic group in which the diploid lineages share a common ancestor with Artemia urmiana while tetraploids are closely related to Artemia sinica. The triploid and pentaploid linages are likely to be directly derived from diploid and tetraploid parthenogens, respectively. Subsequently, west Asia is origin for di‐/triploids, and tetra‐/pentaploids rose from East Asia.     

Unphysical nucleation of diamond in the extended cutoff Tersoff potential

In simulations of carbon materials it is common practice to view the coefficients of the cutoff function as free parameters which can be optimised according to the system of interest. This work examines a common modification to the widely used Tersoff potential in which the coefficient of the upper cutoff is increased to improve the properties of amorphous carbon. Using molecular dynamics simulations, we show that this so-called extended cutoff Tersoff model leads to nucleation of diamond nanocrystals during annealing of amorphous carbon. By varying the density of the system, and examining the radial distribution function in conjunction with the modified cutoff function, we demonstrate that this behaviour is unphysical, and does not represent a new pathway for synthesising diamond. Viewed from a broader perspective, this observation provides a cautionary tale against altering the parameters of empirical potentials without fully considering the wider implications.