Molecular Mechanism of SARS-CoVs Orf6 Targeting the Rae1–Nup98 Complex to Compete With mRNA Nuclear Export

The accessory protein Orf6 is uniquely expressed in sarbecoviruses including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which is an ongoing pandemic. SARS-CoV-2 Orf6 antagonizes host interferon signaling by inhibition of mRNA nuclear export through its interactions with the ribonucleic acid export 1 (Rae1)–nucleoporin 98 (Nup98) complex. Here, we confirmed the direct tight binding of Orf6 to the Rae1-Nup98 complex, which competitively inhibits RNA binding. We determined the crystal structures of both SARS-CoV-2 and SARS-CoV-1 Orf6 C-termini in complex with the Rae1–Nup98 heterodimer. In each structure, SARS-CoV Orf6 occupies the same potential mRNA-binding groove of the Rae1–Nup98 complex, comparable to the previously reported structures of other viral proteins complexed with Rae1-Nup98, indicating that the Rae1–Nup98 complex is a common target for different viruses to impair the nuclear export pathway. Structural analysis and biochemical studies highlight the critical role of the highly conserved methionine (M58) of SARS-CoVs Orf6. Altogether our data unravel a mechanistic understanding of SARS-CoVs Orf6 targeting the mRNA-binding site of the Rae1–Nup98 complex to compete with the nuclear export of host mRNA, which further emphasizes that Orf6 is a critical virulence factor of SARS-CoVs.

Identification of Hotspot Mutations in the N Gene of SARS-CoV-2 in Russian Clinical Samples That May Affect the Detection by Reverse Transcription-PCR

Sensitive and reliable diagnostic test systems based on real-time PCR are of great importance in the fight against the ongoing SARS-CoV-2 pandemic. The genetic variability of the SARS-CoV-2 virus leads to the accumulation of mutations, some of which may affect the sensitivity of modern PCR assays. The aim of this study was to search in Russian clinical samples for new mutations in SARS-CoV-2 gene N that can affect the detection by RT-PCR. In this study, the polymorphisms in the regions of the target gene N causing failed or poor detection of the target N in the RT-PCR assay on 12 selected samples were detected. Sequencing the entire N and E genes in these samples along with other 195 samples that were positive for both target regions was performed. Here, we identified a number of nonsynonymous mutations and one novel deletion in the N gene that affected the ability to detect a target in the N gene as well a few mutations in the E gene of SARS-CoV-2 that did not affect detection. Sequencing revealed that majority of the mutations in the N gene were located in the variable region between positions 193 and 235 aa, inside and nearby the phosphorylated serine-rich region of the protein N. This study highlights the importance of the further characterization of the genetic variability and evolution of gene N, the most common target for detecting SARS-CoV-2. The use of at least two targets for detecting SARS-CoV-2, including one for the E gene, will be necessary for reliable diagnostics.

Exploitation of the Host Ubiquitin System: Means by Legionella pneumophila

Ubiquitination is a commonly used post-translational modification (PTM) in eukaryotic cells, which regulates a wide variety of cellular processes, such as differentiation, apoptosis, cell cycle, and immunity. Because of its essential role in immunity, the ubiquitin network is a common target of infectious agents, which have evolved various effective strategies to hijack and co-opt ubiquitin signaling for their benefit. The intracellular pathogen Legionella pneumophila represents one such example; it utilizes a large cohort of virulence factors called effectors to modulate diverse cellular processes, resulting in the formation a compartment called the Legionella-containing vacuole (LCV) that supports its replication. Many of these effectors function to re-orchestrate ubiquitin signaling with distinct biochemical activities. In this review, we highlight recent progress in the mechanism of action of L. pneumophila effectors involved in ubiquitination and discuss their roles in bacterial virulence and host cell biology.

Neurological Involvement in COVID-19

The respiratory system is the most common target of COVID-19, however, various experimental studies and case reports have shown its affinity for neural tissues. In this chapter, we described pathogenesis and propagation of SARS-CoV-2 virus in the nervous system, potential routes of the SARS-CoV-2 invasion in the brain, as well as indirect effects of COVID-19 on multiorgan disorders. We have also presented all of the reported neurological manifestations in COVID-19 with an explanation of possible underlying pathways. Among patients who tested positive on SARS-CoV-2, various neurological irregularities have been described, affecting both the central and peripheral nervous systems. In general, neurological complications in COVID-19 patients occur within 1 and 14 days, in most cases on average on the 5th day of the incubation period. We have demonstrated all of the reported neurological findings, whereas the most commonly reported were headache, dizziness, myalgia, hypogeusia, hyposmia, and impaired consciousness. More serious neurological conditions in COVID-19 included meningitis, encephalitis, and ischemic or hemorrhagic stroke.

Isolation and Characterisation of Toxic Secondary Metabolites Produced by Black Mould

<p>Microbial secondary metabolites, commonly called natural products, have been crucial for the progression of modern medicine. Not essential for the basic functions of life, secondary metabolites are instead produced to provide a competitive advantage in the environment. The method of action is commonly toxicity to other species in their environment, thereby harming or killing the competition. These toxic properties have allowed them to be utilised as antimicrobial and antitumor agents, however this same toxicity is able to cause detrimental health effects in humans causing symptoms ranging from minor to life threatening. The black mould Stachybotrys chartarum is capable of producing very toxic secondary metabolites called macrocyclic trichothecenes. Satratoxin G (6) and H (7), are two of the most toxic naturally occurring compounds in the world. This has made S. chartarum a common target when adverse health has been associated with damp and mouldy dwellings. However, there is very little evidence for this link beyond its ubiquity and ability to produce the aforementioned highly toxic macrocyclic trichothecenes. This research investigates S. chartarum and the toxic secondary metabolites it produces, with special emphasis on satratoxin G and H. Different culturing methods and resulting morphology are assessed. The satratoxins were isolated from crude extracts and full characterisation by 1-D and 2-D NMR spectroscopy was done. This process revealed differences from the accepted literature, and spectra are reported herein to aid in future identification. The importance of genetics and the public health implications of mould contamination are also discussed.</p>

Isolation and Characterisation of Toxic Secondary Metabolites Produced by Black Mould

<p>Microbial secondary metabolites, commonly called natural products, have been crucial for the progression of modern medicine. Not essential for the basic functions of life, secondary metabolites are instead produced to provide a competitive advantage in the environment. The method of action is commonly toxicity to other species in their environment, thereby harming or killing the competition. These toxic properties have allowed them to be utilised as antimicrobial and antitumor agents, however this same toxicity is able to cause detrimental health effects in humans causing symptoms ranging from minor to life threatening. The black mould Stachybotrys chartarum is capable of producing very toxic secondary metabolites called macrocyclic trichothecenes. Satratoxin G (6) and H (7), are two of the most toxic naturally occurring compounds in the world. This has made S. chartarum a common target when adverse health has been associated with damp and mouldy dwellings. However, there is very little evidence for this link beyond its ubiquity and ability to produce the aforementioned highly toxic macrocyclic trichothecenes. This research investigates S. chartarum and the toxic secondary metabolites it produces, with special emphasis on satratoxin G and H. Different culturing methods and resulting morphology are assessed. The satratoxins were isolated from crude extracts and full characterisation by 1-D and 2-D NMR spectroscopy was done. This process revealed differences from the accepted literature, and spectra are reported herein to aid in future identification. The importance of genetics and the public health implications of mould contamination are also discussed.</p>

Review on Coexistence of Hypertension with Diabetes Mellitus

The coexistence of both diabetes mellitus and hypertension affect the some major target organs. Their common target organ is heart and kidney. The primary goal in the management of the hypertensive diabetic patients is lowering blood pressure to less than 130/80mm Hg Beta- blockers have been reported to adversely affect the overall risk factor profile in the diabetic patient. Initially ACE inhibitors and ARB are initially can be given to diabetic hypertensive. Beta blockers also show great effects in preventing further cardiovascular diseases in diabetic hypertensive. Although combined drug therapy is usually required to achieve goal but in addition to drug therapy some other precautions should also plays effective role like exercise، low sodium chloride intake, lower lipids in diet, maintaining glucose level, stress less patients environment. Calcium channels blockers and diuretics in combination with ACE inhibitors and antidiabetic drugs will also exerts beneficial effects.

Molecular Mechanisms and Cellular Contribution from Lung Fibrosis to Lung Cancer Development

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrosing interstitial lung disease (ILD) of unknown aetiology, with a median survival of 2–4 years from the time of diagnosis. Although IPF has unknown aetiology by definition, there have been identified several risks factors increasing the probability of the onset and progression of the disease in IPF patients such as cigarette smoking and environmental risk factors associated with domestic and occupational exposure. Among them, cigarette smoking together with concomitant emphysema might predispose IPF patients to lung cancer (LC), mostly to non-small cell lung cancer (NSCLC), increasing the risk of lung cancer development. To this purpose, IPF and LC share several cellular and molecular processes driving the progression of both pathologies such as fibroblast transition proliferation and activation, endoplasmic reticulum stress, oxidative stress, and many genetic and epigenetic markers that predispose IPF patients to LC development. Nintedanib, a tyrosine–kinase inhibitor, was firstly developed as an anticancer drug and then recognized as an anti-fibrotic agent based on the common target molecular pathway. In this review our aim is to describe the updated studies on common cellular and molecular mechanisms between IPF and lung cancer, knowledge of which might help to find novel therapeutic targets for this disease combination.

Development and Clinical Validation of CT-Based Regional Centiloid Method for Amyloid PET

Background: We developed and validated CT-based regional Centiloid. A CT-based regional Centiloid was developed and validated in the present study. Methods: For development ofMRI-based or CT-based regional CLs,the cohort consist of 63 subjects (20 young controls (YC) and18 old controls (OC), and 25 Alzheimer’s disease dementia (ADD)).We used direct comparison of FMM-FBB CL (dcCL) method using MRI and CT images to define a common target region and six regional VOIs including the frontal, temporal, parietal, posterior cingulate, occipital and striatal regions. Global and regional dcCL scales were compared between MRI-based and CT-based methods. For clinical validation, cohortconsisted of 2,245subjects (627 in CN, 933 in MCI, and 685 in ADD). Results: Both MRI-based and CT-based dcCL scales showed that FMM and FBB were highly correlated with each other, globally and regionally (R2 = 0.96~0.99). Both FMM and FBB showed that CT-based regional dcCL scales were highly correlated with MRI-based regional dcCL scales (R2 = 0.97~0.99). Absolute differences in regional CL scales between CT-based and MRI-based methods seemed to be relatively insignificant (p>0.05). In our clinical validation study, the G(-)R(+) and G(+)Str(+) groups predict worse neuropsychological performance than the G(-)R(-) and the G(+)Str(-) groups (p< 0.05) respectively.Conclusions: Our findings suggested that it is feasible to convert FMM or FBB dcSUVR values into the dcCL scales regionally without additional MRI scans, which might in turn become a more easily accessible method for researchers and be applicable to a variety of different conditions.

MiR-188-5p and MiR-141-3p Promote Bladder Cancer Synergistically via Targeting PTEN to Activate AKT/c-MYC Signal Pathway

MicroRNAs play a key role in the progress of bladder cancer (BC), which may lead to poor prognosis. A single MicroRNA can be used as an independent factor to regulate the progress of BC, while two MicroRNAs can have a synergistic regulatory effect on BC progress. It has been confirmed in our previous studies that miR-188-5p and miR-141-3p demonstrated high expressions in BC tissues and cells, which can promote the progress of BC and affect patient’s prognosis. As a follow-up research, this study has made further exploration in many aspects. We predicted and confirmed that miR-188-5p and miR-141-3p had a common target gene PTEN which had low expression in BC tissues and cells. Down regulation of PTEN can promote the progress of BC, and significantly reverse the inhibitory effect of down-regulated miR-188-5p and miR-141-3p on BC progress. PTEN is a cancer suppressor gene. Experiments further verified that pAKT and c-MYC were downstream effector proteins of PTEN, and their expressions increased with the decrease of PTEN expression. Experiments manifested that down-regulating miR-188-5p and miR-141-3p could significantly inhibit the volume and weight of subcutaneous tumors in mice, and half dose co-transfection of the two miRs made the tumor smaller and lighter. Therefore, it was concluded that miR-188-5p and miR-141-3p promoted the progress of BC synergistically by inhibiting PTEN to activate AKT/c-MYC pathway.