Comparative evaluation of hydro-, chemo- , and hormonal-priming methods for imparting salt and PEG stress tolerance in Indian mustard (<Emphasis Type="Italic">Brassica</Emphasis><Emphasis Type="Italic">juncea</Emphasis> L.)

The present study was aimed to evaluate the effect of different seed priming methods to enhance the sodium chloride (NaCl) and polyethylene glycol-8000 (PEG-8000) stress tolerance in Indian mustard (Brassica juncea L.). Seeds subjected to different priming treatments such as water (hydro-priming), calcium chloride (CaCl₂) (chemo-priming), and abscisic acid (ABA) (hormonal-priming) showed increased rate of germination as compared to non-primed seeds. The primed and non-primed seeds were grown for 15 days and then the seedlings were independently subjected to iso-osmotic salt (150 mM NaCl) or PEG-8000 (20%) stress. The different biochemical responses were studied 10 days after treatment. Under NaCl and PEG stress, the dry weight and total chlorophyll content were higher in primed sets as compared to non-primed treatment which was also evident by the phenotype of the seedlings. In general, the higher activities of superoxide dismutase and glutathione reductase resulted in lower oxidative damage, in terms of malondialdehyde content, under NaCl and PEG stress in hydro-primed set as compared to non-primed, ABA-, and CaCl₂-primed treatments. Besides, the level of total phenolics and accumulation of osmolytes such as free proline, glycine betaine, and total soluble sugars was also lower in hydro-primed set as compared to other primed and non-primed treatments. The study thus suggests the use of hydro-priming as a simple and cost-effective strategy to alleviate the NaCl and PEG induced stress in B. juncea.     

Colon Delivery of Budesonide: Evaluation of Chitosan–Chondroitin Sulfate Interpolymer Complex

The present study was aimed at formulating tablets comprising of coating susceptible to microbial enzyme degradation for releasing budesonide in the colon. Tablets prepared by using Avicel® pH 102 as diluent and Eudragit® L100-55 as binder were coated to a weight gain of 10% w/w employing aqueous mixtures containing chitosan (CH) and chondroitin sulfate (CS). The interpolymer complex between CH and CS was characterized using Fourier transform infrared (FTIR) and differential scanning calorimetery (DSC) studies. The tablets were evaluated for release of budesonide through in vitro in vivo studies. Formation of bonds between –COO^? and –OSO ₃ ^? groups of CS and –NH ₃ ⁺ groups of CH was evident in the FTIR spectra of these interpolymer complexed (IPC) films. The DSC thermograms of these films revealed one endothermic transition between 190°C and 205°C, suggesting the formation of new bonds in the IPC. The pH sensitive swelling exhibited by these films was observed to be a function of CH concentration. Tablets coated with aqueous mixtures containing 40:60 or 50:50 ratio of CH/CS totally prevented the release of budesonide in pH 1.2 buffer. The peaks (FTIR) and endothermic transitions (DSC) characteristic of interpolymer complexation were observed to remain unaffected after sequential exposure of the films to pH 1.2 and pH 7.4 buffer IP. This proved the versatility of these IPC films for colon delivery. C _max of 1,168.99 and 1,174.2 ng/mL, respectively, at 12 and 8 h post-oral dosing of tablets coated with 40:60 or 50:50 ratio of CH/CS was observed in rats. The aqueous CH/CS (40:60) coating could provide a facile method for delivering budesonide to the colon.     

E(z), a depth-dependent potential for assessing the energies of insertion of amino acid side-chains into membranes: derivation and applications to determining the orientation of transmembrane and interfacial helices

We have developed an empirical residue-based potential (E(z) potential) for protein insertion in lipid membranes. Propensities for occurrence as a function of depth in the bilayer were calculated for the individual amino acid types from their distribution in known structures of helical membrane proteins. The propensities were then fit to continuous curves and converted to a potential using a reverse-Boltzman relationship. The E(z) potential demonstrated a good correlation with experimental data such as amino acid transfer free energy scales (water to membrane center and water to interface), and it incorporates transmembrane helices of varying composition in the membrane with trends similar to those obtained with translocon-mediated insertion experiments. The potential has a variety of applications in the analysis of natural membrane proteins as well as in the design of new ones. It can help in calculating the propensity of single helices to insert in the bilayer and estimate their tilt angle with respect to the bilayer normal. It can be utilized to discriminate amphiphilic helices that assume a parallel orientation at the membrane interface, such as those of membrane-active peptides. In membrane protein design applications, the potential allows an environment-dependent selection of amino acid identities.     

Development of a new mouse model of acute pancreatitis induced by administration of L-arginine

The pathogenesis of acute pancreatitis is not fully understood. Experimental animal models that mimic human disease are essential to better understand the pathophysiology of the disease and to evaluate potential therapeutic agents. Given that the mouse genome is known completely and that a large number of strains with various genetic deletions are available, it is advantageous to have multiple reliable mouse models of acute pancreatitis. Presently, there is only one predominant model of acute pancreatitis in mice, in which hyperstimulatory doses of cholecystokinin or its analog caerulein are administered. Therefore, the aim of this study was to develop another mouse model of acute pancreatitis. In this study, C57BL/6 mice were injected intraperitoneally with L-arginine in two doses of 4 g/kg each, 1 h apart. Serum amylase, myeloperoxidase, and histopathology were examined at varying time points after injection to assess injury to the pancreas and lung. We found that injection of L-arginine was followed by significant increases in plasma amylase and pancreatic myeloperoxidase accompanied by marked histopathological changes. The injury to the pancreas was slow to develop and peaked at 72 h. Subsequent to peak injury, the damaged areas contained collagen fibers as assessed by increased Sirius red staining. In contrast, D-arginine or other amino acids did not cause injury to the pancreas. In addition, acute inflammation in the pancreas was associated with lung injury. Our results indicate that administration of L-arginine to mice results in severe acute pancreatitis. This model should help in elucidating the pathophysiology of pancreatitis.     

Plant mitochondrial recombination surveillance requires unusual RecA and MutS homologs

For >20 years, the enigmatic behavior of plant mitochondrial genomes has been well described but not well understood. Chimeric genes appear, and occasionally are differentially replicated or expressed, with significant effects on plant phenotype, most notably on male fertility, yet the mechanisms of DNA replication, chimera formation, and recombination have remained elusive. Using mutations in two important genes of mitochondrial DNA metabolism, we have observed reproducible asymmetric recombination events occurring at specific locations in the mitochondrial genome. Based on these experiments and existing models of double-strand break repair, we propose a model for plant mitochondrial DNA replication, chimeric gene formation, and the illegitimate recombination events that lead to stoichiometric changes. We also address the physiological and developmental effects of aberrant events in mitochondrial genome maintenance, showing that mitochondrial genome rearrangements, when controlled, influence plant reproduction, but when uncontrolled, lead to aberrant growth phenotypes and dramatic reduction of the cell cycle.     

Pulmonary Artery Pseudoaneurysm in COVID-19-Associated Pulmonary Mucormycosis: Case Series and Systematic Review of the Literature

Mycopathologia - Literature on COVID-19-associated pulmonary mucormycosis (CAPM) is sparse. Pulmonary artery pseudoaneurysm (PAP) is an uncommon complication of pulmonary mucormycosis (PM), and...     

Allostery mediates ligand binding to WWOX tumor suppressor via a conformational switch

While being devoid of the ability to recognize ligands itself, the WW2 domain is believed to aid ligand binding to the WW1 domain in the context of a WW1–WW2 tandem module of WW domain‐containing oxidoreductase (WWOX) tumor suppressor. In an effort to test the generality of this hypothesis, we have undertaken here a detailed biophysical analysis of the binding of WW domains of WWOX alone and in the context of the WW1–WW2 tandem module to an array of putative proline‐proline‐x–tyrosine (PPXY) ligands. Our data show that while the WW1 domain of WWOX binds to all ligands in a physiologically relevant manner, the WW2 domain does not. Moreover, ligand binding to the WW1 domain in the context of the WW1–WW2 tandem module is two‐to‐three‐fold stronger than when treated alone. We also provide evidence that the WW domains within the WW1–WW2 tandem module physically associate so as to adopt a fixed spatial orientation relative to each other. Of particular note is the observation that the physical association of the WW2 domain with WW1 blocks access to ligands. Consequently, ligand binding to the WW1 domain not only results in the displacement of the WW2 lid but also disrupts the physical association of WW domains in the liganded conformation. Taken together, our study underscores a key role of allosteric communication in the ability of the WW2 orphan domain to chaperone physiological action of the WW1 domain within the context of the WW1–WW2 tandem module of WWOX. Copyright © 2015 John Wiley & Sons, Ltd.