Aspects of tree shrew consolidated sleep structure resemble human sleep

Understanding human sleep requires appropriate animal models. Sleep has been extensively studied in rodents, although rodent sleep differs substantially from human sleep. Here we investigate sleep in tree shrews, small diurnal mammals phylogenetically close to primates, and compare it to sleep in rats and humans using electrophysiological recordings from frontal cortex of each species. Tree shrews exhibited consolidated sleep, with a sleep bout duration parameter, τ, uncharacteristically high for a small mammal, and differing substantially from the sleep of rodents that is often punctuated by wakefulness. Two NREM sleep stages were observed in tree shrews: NREM, characterized by high delta waves and spindles, and an intermediate stage (IS-NREM) occurring on NREM to REM transitions and consisting of intermediate delta waves with concomitant theta-alpha activity. While IS-NREM activity was reliable in tree shrews, we could also detect it in human EEG data, on a subset of transitions. Finally, coupling events between sleep spindles and slow waves clustered near the beginning of the sleep period in tree shrews, paralleling humans, whereas they were more evenly distributed in rats. Our results suggest considerable homology of sleep structure between humans and tree shrews despite the large difference in body mass between these species.

Over-Expression, Purification, and Characterization of Plant LeGST01, the Mammalian GST-Omega Homolog that Interacts with LeTPX1 from a Tomato cDNA Library

The study focuses on an in vivo GST- omega homologue (pCRT7/TPxII intB4) over-expression, purification and characterization. Experiments purport to characterize the antioxidant activity of the LeTPx1, the interacting glutathione S-transferases BI-GST/GPx, LeGST-T1, T2, T3, T4, T5 and the mammalian inhibitor of apoptosis Bcl-2. Upon specific expression, the proteins exerted, differential protective effects in yeast cells treated with lethal doses of the prooxidants hydrogen peroxide, t-butyl hydroperoxide, and cumene hydroperoxide. The antioxidant activity of LeTPx1 was highest against the cumene hydroperoxide. The overexpressing GST (omega) homologue TPxintB4 (Baier and Dietz, 1999) which share a considerable homology of the mammalian GST-omega1. In conclusion, the work shows that yeast parental strains are extremely sensitive to very low concentrations (0.2mM) of Cumenehydroperoxide. However, after applying the different antioxidants; it appears that the smallest concentrations t to be tolerated.

Arabidopsis Transmembrane Receptor-Like Kinases (RLKs): A Bridge between Extracellular Signal and Intracellular Regulatory Machinery

Receptors form the crux for any biochemical signaling. Receptor-like kinases (RLKs) are conserved protein kinases in eukaryotes that establish signaling circuits to transduce information from outer plant cell membrane to the nucleus of plant cells, eventually activating processes directing growth, development, stress responses, and disease resistance. Plant RLKs share considerable homology with the receptor tyrosine kinases (RTKs) of the animal system, differing at the site of phosphorylation. Typically, RLKs have a membrane-localization signal in the amino-terminal, followed by an extracellular ligand-binding domain, a solitary membrane-spanning domain, and a cytoplasmic kinase domain. The functional characterization of ligand-binding domains of the various RLKs has demonstrated their essential role in the perception of extracellular stimuli, while its cytosolic kinase domain is usually confined to the phosphorylation of their substrates to control downstream regulatory machinery. Identification of the several ligands of RLKs, as well as a few of its immediate substrates have predominantly contributed to a better understanding of the fundamental signaling mechanisms. In the model plant Arabidopsis, several studies have indicated that multiple RLKs are involved in modulating various types of physiological roles via diverse signaling routes. Here, we summarize recent advances and provide an updated overview of transmembrane RLKs in Arabidopsis.

Morphological Description, Phylogenetic and Molecular Analysis of Dirofilaria immitis Isolated from Dogs in the Northwest of Iran

Background: Dirofilariasis is a globally distributed arthropod-borne parasitic disease of mainly canids and felids. We evaluated to extend the knowledge of morpho-molecular characteristics and outer ultrastructure of Dirofilaria immitis isolated from Northwest of Iran. Methods: Overall, 67 filarial worms including 26 males parasites were collected from the cardiovascular system of the 43 stray dogs in Meshkinshar, Ardebil Province, Northwest of Iran in 2017, and subjected to light and scanning electron microscopy (SEM) as well as carmine alum staining for morpho-molecular and identification. Molecular methods were used for confirmation of morphological findings by sequencing of Cytochrome c oxidase subunit I (cox1) gene. Results: The partial DNA sequencing of cox1 gene of adult parasites showed considerable homology and close proximity to the previously isolated from Kerman and Meshkinshahr, Iran. The lowest genetic variation and the highest intra-species variability was found in D. immitis and D. repens, respectively. No similarity was identified between D. immitis nucleotide sequence and Wolbachia species as its endosymbiont bacteria. Conclusion: The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.

The N-glycans of Chlorella sorokiniana and a related strain contain arabinose but have strikingly different structures

The many emerging applications of microalgae such as Chlorella also instigate interest in their ability to conduct protein modifications such as N-glycosylation. Chlorella vulgaris has recently been shown to equip its proteins with highly O-methylated oligomannosidic N-glycans. Two other frequently occurring species names are Chlorella sorokiniana and Chlorella pyrenoidosa—even though the latter is taxonomically ill defined. We analyzed by mass spectrometry and nuclear magnetic resonance spectroscopy the N-glycans of type culture collection strains of C. sorokiniana and of a commercial product labeled C. pyrenoidosa. Both samples contained arabinose, which has hitherto not been found in N-glycans. Apart from this only commonality, the structures differed fundamentally from each other and from that of N-glycans of land plants. Despite these differences, the two algae lines exhibited considerable homology in their ITS1-5.8S-ITS2 rDNA sequences. These drastic differences of N-glycan structures between species belonging to the very same genus provoke questions as to the biological function on a unicellular organism.

Identification of a Novel Invasion-Promoting Region in Insulin Receptor Substrate 2

ABSTRACT Although the insulin receptor substrate (IRS) proteins IRS1 and IRS2 share considerable homology and activate common signaling pathways, their contributions to breast cancer are distinct. IRS1 has been implicated in the proliferation and survival of breast tumor cells. In contrast, IRS2 facilitates glycolysis, invasion, and metastasis. To determine the mechanistic basis for IRS2-dependent functions, we investigated unique structural features of IRS2 that are required for invasion. Our studies revealed that the ability of IRS2 to promote invasion is dependent upon upstream insulin-like growth factor 1 receptor (IGF-1R)/insulin receptor (IR) activation and the recruitment and activation of phosphatidylinositol 3-kinase (PI3K), functions shared with IRS1. In addition, a 174-amino-acid region in the IRS2 C-terminal tail, which is not conserved in IRS1, is also required for IRS2-mediated invasion. Importantly, this “invasion (INV) region” is sufficient to confer invasion-promoting ability when swapped into IRS1. However, the INV region is not required for the IRS2-dependent regulation of glucose uptake. Bone morphogenetic protein 2-inducible kinase (BMP2K) binds to the INV region and contributes to IRS2-dependent invasion. Taken together, our data advance the mechanistic understanding of how IRS2 regulates invasion and reveal that IRS2 functions important for cancer can be independently targeted without interfering with the metabolic activities of this adaptor protein.

Computational Analysis of Dengue and Chikungunya Viral Gene Sequences To Elucidate Their Transmission and/or Symptoms Similarity

Dengue (DENV) and Chikungunya (CHIKV) viruses are enveloped positive-strand RNA mosquito-borne viruses that have been disseminated worldwide. Both the viruses are dissimilar in many aspects like genome organization and expression, particle size and taxonomical status except two similar aspects i.e. mode of transmission and the disease pathogenecity. Reports are indicating the co-infection of the both the viruses in a same patient. In view of similarities and/or dissimilarities between these two viruses, we indented to identify the existence of homology between any two gene sequences of which one from each virus. For this study we have analyzed all the structural and non structural nucleotide sequences of DENV type 1 (ACCESSION no. NC_001477) and CHIKV IND-06-Guj strain (ACCESSION no. JF272477) using the computational techniques. The results indicated that there is no homology between structural genes of both viruses. But analysis of the non-structural genes revealed that DENV NS4B and CHIKV nsp2 is showing the considerable homology. It is reported that DENV NS4B is involved in maintaining the replication balance in mosquito and human hosts. Hence we predict based on our results, CHIKV nsp2 which is showing homology with the DENV NS4B also serving same purpose in case of CHIKV. Further functional analysis of DENV NS4B and CHIKV nsP2 revealed the information that both have role in IFN inhibition and likely to have an important role in viral pathogenesis. Identification of common or shared epitopes on these polypeptides could lead to the development of an immunogen which could serve as vaccine candidate for both the viruses.

Screening of Predicted Secreted Antigens from Mycobacterium bovis Reveals the Immunodominance of the ESAT-6 Protein Family

ABSTRACT Results of previous studies utilizing bioinformatic approaches in antigen-mining experiments revealed that secreted proteins are among the most frequently recognized antigens from Mycobacterium bovis. Thus, we hypothesized that the analysis of secreted proteins is likely to reveal additional immunogenic antigens that can be used to increase the specificity of diagnostic tests or be suitable vaccination candidates for mycobacterial infections. To test this hypothesis, 382 pools of overlapping peptides spanning 119 M. bovis secreted and potentially secreted proteins were screened for the ability to stimulate a gamma interferon response in vitro using whole blood from tuberculin-positive reactor (TB reactor) cattle. Of the 119 proteins screened, 70 (59%) induced positive responses in the TB reactor animals to various degrees. Strikingly, all but one of the 15 ESAT-6 proteins tested were recognized by at least 30% of the TB reactor animals, with 12 of the 22 most commonly recognized antigens belonging to this protein family. Further analysis of these data demonstrated that there was no significant difference in immunogenicity between the ESAT-6 proteins that were components of potentially intact ESX secretory systems and those corresponding to additional partial esx loci. Importantly for vaccine design, antigenic epitopes in some highly conserved regions shared by numerous ESAT-6 proteins were identified. However, despite this considerable homology, peptide-mapping experiments also revealed that immunodominant peptides were located in regions of amino acid variability.

Circulating markers for endocrine tumours of the gastroenteropancreatic tract

The diffuse endocrine system (DES) includes a wide range of secretory cells that may be the source of tumours. Gastroenteropancreatic endocrine (GEP) tumours arising within the DES secrete a variety of peptides and amines that are found in the circulation and are responsible for the syndromes associated with these tumours. In this review, the most common tumours of the GEP tract are outlined and the circulating products of these tumours identified. Where differential diagnosis is difficult these points are addressed. The peptides most commonly secreted by GEP neuroendocrine tumours are identified and described and their biological activities are discussed. Current methods available for measurement of these peptides are described. Attention is drawn towards molecular specificity where appropriate, as many pancreatic and gut peptides fall within families which show considerable homology, such as the tachykinin family or the glucagon family. Other peptides such as gastrin circulate in multiple molecular forms. This homology and diversity may cause difficulty in the interpretation of peptide measurements in the clinical situation if assays are not specific.

ACEH/ACE2 is a novel mammalian metallocarboxypeptidase and a homologue of angiotensin-converting enzyme insensitive to ACE inhibitors

A human zinc metalloprotease (termed ACEH or ACE2) with considerable homology to angiotensin- converting enzyme (ACE) (EC 3.4.15.1) has been identified and subsequently cloned and functionally expressed. The translated protein contains an N-terminal signal sequence, a single catalytic domain with zinc-binding motif (HEMGH), a transmembrane region, and a small C-terminal cytosolic domain. Unlike somatic ACE, ACEH functions as a carboxypeptidase when acting on angiotensin I and angiotensin II or other peptide substrates. ACEH may function in conjunction with ACE and neprilysin in novel pathways of angiotensin metabolism of physiological significance. In contrast with ACE, ACEH does not hydrolyse bradykinin and is not inhibited by typical ACE inhibitors. ACEH is unique among mammalian carboxypeptidases in containing an HEXXH zinc motif but, in this respect, resembles a bacterial enzyme, Thermus aquaticus (Taq) carboxypeptidase (EC 3.4.17.19). Collectrin, a developmentally regulated renal protein, is homologous with the C-terminal region of ACEH but has no similarity with ACE and no catalytic domain. Thus, the ACEH protein may have evolved as a chimera of a single ACE-like domain and a collectrin domain. The collectrin domain may regulate tissue response to injury whereas the catalytic domain is involved in peptide processing events.Key words: ACEH, ACE2, metalloprotease, collectrin, carboxypeptidase, angiotensin II.