Comparative transcriptome analysis provides insight into the important pathways and key genes related to the pollen abortion in the thermo-sensitive genic male sterile line 373S in Brassica napus L.

The thermo-sensitive genic male sterility (TGMS) system plays a key role in the production of two-line hybrids in rapeseed (Brassica napus). To uncover key cellular events and genetic regulation associated with TGMS, a combined study using cytological methods and RNA-sequencing analysis was conducted for the rapeseed TGMS line 373S. Cytological studies showed that microspore cytoplasm of 373S plants was condensed, the microspore nucleus was degraded at an early stage, the exine was irregular, and the tapetum developed abnormally, eventually leading to male sterility. RNA-sequencing analysis identified 430 differentially expressed genes (298 upregulated and 132 downregulated) between the fertile and sterile samples. Gene ontology analysis demonstrated that the most highly represented biological processes included sporopollenin biosynthetic process, pollen exine formation, and extracellular matrix assembly. Kyoto encyclopedia of genes and genomes analysis indicated that the enriched pathways included amino acid metabolism, carbohydrate metabolism, and lipid metabolism. Moreover, 26 transcript factors were identified, which may be associated with abnormal tapetum degeneration and exine formation. Subsequently, 19 key genes were selected, which are considered to regulate pollen development and even participate in pollen exine formation. Our results will provide important insight into the molecular mechanisms underlying TGMS in rapeseed.

Human Cytomegalovirus Induced Aberrant Expression of Non-coding RNAs.

Human cytomegalovirus (HCMV) is a ß-herpesvirus whose genome consists of double stranded linear DNA. HCMV genome can generate non-coding RNAs (ncRNAs) through transcription in its host cells. Besides that, HCMV infection also changes the ncRNAs expression profile of the host cells. ncRNAs play a key role in maintaining the normal physiological activity of cells, and the disorder of ncRNAs expression has numerous adverse effects on cells. However, until now, the relationship between ncRNAs and HCMV-induced adverse effects are not summarized in detail. This review aims to give a systematic summary of the role of HCMV infection in ncRNAs expression while providing insights into the molecular mechanism of unnormal cellular events caused by ncRNAs disorder. ncRNAs disorder induced by HCMV infection is highly associated with cell proliferation, apoptosis, tumorigenesis, and immune regulation, as well as the development of cardiovascular diseases, and the potential role of biomarker. We summarize the studies on HCMV associated ncRNAs disorder and suggest innovative strategies for eliminating the adverse effects caused by HCMV infection.

Signaling Modulation by miRNA-221-3p During Tooth Morphogenesis in Mice.

miRNAs are conserved short non-coding RNAs that play a role in the modulation of various biological pathways during tissue and organ morphogenesis. In this study, the function of miRNA-221-3p in tooth development, through its loss or gain in function was evaluated. A variety of techniques were utilized to evaluate detailed functional roles of miRNA-221-3p during odontogenesis, including in vitro tooth cultivation, renal capsule transplantation, in situ hybridization, real-time PCR, and immunohistochemistry. Two-day in vitro tooth cultivation at E13 identified altered cellular events, including cellular proliferation, apoptosis, adhesion, and cytoskeletal arrangement, with the loss and gain of miRNA-221-3p. qPCR analysis revealed alterations in gene expression of tooth-related signaling molecules, including ß-catenin, Bmp2, Bmp4, Fgf4, Ptch1, and Shh, when inhibited with miRNA-221-3p and mimic. Also, the inhibition of miRNA-221-3p demonstrated increased mesenchymal localizations of pSMAD1/5/8, alongside decreased expression patterns of Shh and Fgf4 within inner enamel epithelium (IEE) in E13 + 2 days in vitro cultivated teeth. Moreover, 1-week renal transplantation of in vitro cultivated teeth had smaller tooth size with reduced enamel and dentin matrices, along with increased cellular proliferation and Shh expression along the Hertwig epithelial root sheath (HERS), within the inhibitor group. Similarly, in 3-week renal calcified teeth, the overexpression of miRNA-221-3p did not affect tooth phenotype, while the loss of function resulted in long and slender teeth with short mesiodistal length. This study provides evidence that a suitable level of miRNA-221-3p is required for the modulation of major signaling pathways, including Wnt, Bmp, and Shh, during tooth morphogenesis.

How Do Post-Translational Modifications Influence the Pathomechanistic Landscape of Huntington's Disease? A Comprehensive Review.

Huntington's disease (HD) is an autosomal dominant inherited neurodegenerative disorder characterized by the loss of motor control and cognitive ability, which eventually leads to death. The mutant huntingtin protein (HTT) exhibits an expansion of a polyglutamine repeat. The mechanism of pathogenesis is still not fully characterized; however, evidence suggests that post-translational modifications (PTMs) of HTT and upstream and downstream proteins of neuronal signaling pathways are involved. The determination and characterization of PTMs are essential to understand the mechanisms at work in HD, to define possible therapeutic targets better, and to challenge the scientific community to develop new approaches and methods. The discovery and characterization of a panoply of PTMs in HTT aggregation and cellular events in HD will bring us closer to understanding how the expression of mutant polyglutamine-containing HTT affects cellular homeostasis that leads to the perturbation of cell functions, neurotoxicity, and finally, cell death. Hence, here we review the current knowledge on recently identified PTMs of HD-related proteins and their pathophysiological relevance in the formation of abnormal protein aggregates, proteolytic dysfunction, and alterations of mitochondrial and metabolic pathways, neuroinflammatory regulation, excitotoxicity, and abnormal regulation of gene expression.

Review of Emerging Japanese Encephalitis Virus: New Aspects and Concepts about Entry into the Brain and Inter-Cellular Spreading.

Japanese encephalitis virus (JEV) is an emerging flavivirus of the Asia-Pacific region. More than two billion people live in endemic or epidemic areas and are at risk of infection. Recently, the first autochthonous human case was recorded in Africa, and infected birds have been found in Europe. JEV may spread even further to other continents. The first section of this review covers established and new information about the epidemiology of JEV. The subsequent sections focus on the impact of JEV on humans, including the natural course and immunity. Furthermore, new concepts are discussed about JEV's entry into the brain. Finally, interactions of JEV and host cells are covered, as well as how JEV may spread in the body through latently infected immune cells and cell-to-cell transmission of virions or via other infectious material, including JEV genomic RNA.

Lysyl-tRNA synthetase (Krs) acts a virulence factor of Beauveria bassiana by its vital role in conidial germination and dimorphic transition.

Krs is a class II lysyl-tRNA synthetase (KRS) that is involved in cytosolic protein synthesis in budding yeast but functionally has not been explored in filamentous fungi. Previous transcriptomic analysis has revealed that a Krs-coding gene is likely involved in pathogenesis of Beauveria bassiana, a classic insect pathogen as a global source of fungal insecticides. Here, we show that Krs is localized in the cytoplasm of hyphal cells and acts as a substantial virulence factor in B. bassiana. Deletion of krs resulted in 10-h delayed germination, decreased (15 %) thermotolerance, and lowered (46 %) UV-B resistance of aerial conidia despite limited impact on conidiation capacity and slight or inconspicuous influence on radial growth on rich and minimal media with different carbon (10 sugars/polyols) and nitrogen (17 amino acids) sources. The deletion mutant suffered 58 % reduction in submerged blastospore yield (an index of in vitro dimorphic transition rate) in a minimal medium, and the reduction increased to 71 % in another trehalose-based medium mimic to insect haemolymph. In standardized bioassays, median lethal actions of Δkrs against Galleria mellonella larvae through the infections passing and bypassing the insect cuticle were prolonged to 192 and 153 h from wild-type median lethal time (LT50) estimates of 119 and 109 h, respectively. Microscopic examination revealed 2-d delayed presence of in vivo formed hyphal bodies in the haemolymph of the larvae infected by Δkrs in either mode. These findings unveil a vital role of Krs in conidial germination and dimorphic transition and its contribution to the fungal potential against arthropod pests.

Glial-associated changes in the cerebral cortex after collagenase-induced intracerebral hemorrhage in the rat striatum.

Striatum and the cerebral cortex are regions susceptible to secondary injury after intracerebral hemorrhage (ICH) and glial cells in tissue adjacent to the hematoma may modulate cellular vulnerability after brain damage. Nonetheless, while the glial- associated changes occurring in the cerebral cortex after ICH may be important in maximizing brain recovery, they are not fully understood. The aim of this study was to evaluate the temporal profile of glial-associated changes in the cerebral cortex after ICH. First, the motor consequences of ICH and its relation to the lesion volume were analyzed. Secondly, glial cell proportion (GFAP+ and S100B+ astrocytes, CD11+ microglia) in the ipsilesional sensorimotor cortex and striatum, using flow cytometry were evaluated. ELISA was used to measure GFAP and S100B content in these structures as well as S100B levels in serum and cerebral spinal fluid. Main results revealed that ICH induced a delayed increase in GFAP+ cells in the sensorimotor cortex, as compared to the striatum, although the pattern of GFAP expression was similar in both structures. Interestingly, the time-curve patterns of both S100B and CD11+ microglial cells differed between the cortex and striatum. Altogether, these results suggest a different dynamics of glial-associated changes in the cerebral cortex, suggesting it is a vulnerable structure and undergoes an independent secondary process of reactive glial plasticity following intracerebral hemorrhage.

Molecular and biological pathways of skeletal muscle dysfunction in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) will be a major leading cause of death worldwide in the near future. Weakness and atrophy of the quadriceps are associated with a significantly poorer prognosis and increased mortality in COPD. Despite that skeletal muscle dysfunction may affect both respiratory and limb muscle groups in COPD, the latter are frequently more severely affected. Therefore, muscle dysfunction in COPD is a common systemic manifestation that should be evaluated on routine basis in clinical settings. In the present review, several aspects of COPD muscle dysfunction are being reviewed, with special emphasis on the underlying biological mechanisms. Figures on the prevalence of COPD muscle dysfunction and the most relevant etiologic contributors are also provided. Despite that ongoing research will shed light into the contribution of additional mechanisms to COPD muscle dysfunction, current knowledge points toward the involvement of a wide spectrum of cellular and molecular events that are differentially expressed in respiratory and limb muscles. Such mechanisms are thoroughly described in the article. The contribution of epigenetic events on COPD muscle dysfunction is also reviewed. We conclude that in view of the latest discoveries, from now, on new avenues of research should be designed to specifically target cellular mechanisms and pathways that impair muscle mass and function in COPD using pharmacological strategies and/or exercise training modalities.

A proteomic analysis of the early secondary molecular effects caused by Cn2 scorpion toxin on neuroblastoma cells.

Although the primary physiological effects produced by scorpion toxins are already well known, most of the secondary molecular events following scorpion neurotoxins-ion channel interactions are poorly understood and described. For this reason, we used a proteomic approach to determine the changes in relative protein abundance in F11 mouse neuroblastoma cells treated with Cn2, the major ß-toxin from the venom of the scorpion Centruroides noxius Hoffmann. Here we show that the relative abundance of 24 proteins changed after Cn2 treatment. Proteins related to protection from apoptosis and cell survival, as well as those involved in cell morphology and some translation elongation factors were diminished. By contrast, proteins associated with energy metabolism were increased. Additionally, results of western immunoblots confirmed the preference of action towards some special targets. These results suggest that Cn2 could modify the neuronal structure and induce apoptosis and reduction of the proliferation and cell survival. To support this conclusion we directly measured the Cn2 effect on cell proliferation, division and apoptosis. Our results open new avenues for continuing the studies aimed at better understanding the envenomation process caused by scorpion stings. BIOLOGICAL SIGNIFICANCE: The purpose of this work was to identify which proteins, apart from the ion-channels, are involved in the envenomation process in order to develop possible strategies to circumvent the deleterious effects caused by the toxic peptides of the venom. For this reason, we characterized the early changes in the proteome of F11 cells induced by Cn2, the major toxin of the New World scorpion C. noxius Hoffmann, using 2D-PAGE and LC-MS/MS. We identified 24 proteins which relative abundance is modified after the Cn2 treatment. Among these, proteins related with apoptosis protection, cell survival, neuronal morphology and some translation elongation factors were diminished, whereas proteins associated with energy metabolism were increased.