Mutations in the receptor-binding domain of human SARS CoV-2 spike protein increases its affinity to bind human ACE-2 receptor

The severe acute respiratory syndrome virus-2 (SARS CoV-2) infection has resulted in the current global pandemic. The binding of SARS CoV-2 spike protein receptor-binding domain (RBD) to the human angiotensin converting enzyme-2 (ACE-2) receptor causes the host infection. The spike protein has undergone several mutations with reference to the initial strain isolated during December 2019 from Wuhan, China. A number of these mutant strains have been reported as variants of concern and as variants being monitored. Some of these mutants are known to be responsible for increased transmissibility of the virus.
The reason for the increased transmissibility caused by the point mutations can be understood by studying the structural implications and inter-molecular interactions in the binding of viral spike protein RBD and human ACE-2. Here, we use the crystal structure of the RBD in complex with ACE-2 available in www.joplink.net/coronavirus-proteins/ the public domain and analyse the 250 ns molecular dynamics (MD) simulations of wild-type and mutants; K417N, K417T, N440K, N501Y, L452R, T478K, E484K and S494P.
The ionic, hydrophobic and hydrogen bond interactions, amino acid residue flexibility, binding energies and structural variations are characterized. The MD simulations provide clues to the molecular mechanisms of ACE-2 receptor binding in wild-type and mutant complexes. The mutant spike proteins RBD were associated with greater binding affinity with ACE-2 receptor. Communicated by Ramaswamy H. Sarma.

COVID-19 and Alzheimer’s disease: Meninges-mediated neuropathology

SARS-CoV-2 the causative agent of COVID-19 displays a broad range of pathophysiology. Cytokine storms associated with COVID-19 damage the blood-brain barrier (BBB) and allow pro-inflammatory factors to invade the brain, further promoting neurodegeneration. While SARS-CoV-2 viral RNA and proteins have been detected in brain tissues, the mechanisms of neuroinvasion remain unknown. COVID-19 has had a disproportionate impact on those suffering from neurodegenerative disorders such as Alzheimer’s disease (AD).
Understanding the mechanisms of SARS-CoV-2 neuroinvasion is crucial to study the long-term neurocognitive effects of COVID-19 on AD pathology.
Viruses can infiltrate the brain through the meninges via infected immune cells. The meninges regulate the immune surveillance of the brain and play a key role in the efflux of pathogens from the brain. Meningeal dysfunction has been demonstrated to exacerbate amyloid-beta pathogenesis. In this study, we explore the neuroinvasion pathway of SARS-CoV-2 through the meninges and its effect on AD pathology.
Method: 5x FAD x hACE2 mice were inoculated intranasally with a sublethal dose of SARS-CoV-2. The mice were maintained for 2 weeks. Mouse brains and meninges were harvested. The tissue was stained and immunofluorescence imaging was conducted to study viral proliferation and immune responses. Histo-cytometry was conducted for quantitative imaging analysis. Gene expression studies were done using Nanostring assays. All experiments involving the SARS-Cov-2 virus were carried out in a BSL3 facility.
Result: This ongoing study demonstrates the proliferation of the SARS-CoV-2 virus in the brain via meningeal lymphatics. SARS-CoV-2 infection resulted in increased neuroinflammation. Additionally, inflammatory responses induced meningeal dysfunction resulting in increased amyloid-beta pathology and cerebrospinal fluid drainage.
Conclusion: Given the increasing evidence for a viral hypothesis of Alzheimer’s Disease it is extremely important to study the neurodegenerative effects of COVID-19 which has affected millions worldwide. We demonstrate that SARS-CoV-2 infiltrates the brain via the meninges promoting neuroinflammation. Furthermore, amyloid-beta pathologies are exacerbated by COVID-19 in animal models providing preclinical evidence of the long-term neurodegenerative effects of COVID-19.

Exosomes Recovered From the Plasma of COVID-19 Patients Expose SARS-CoV-2 Spike-Derived Fragments and Contribute to the Adaptive Immune Response

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by beta-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has rapidly spread across the globe starting from February 2020. It is well established that during viral infection, extracellular vesicles become delivery/presenting vectors of viral material. However, studies regarding extracellular vesicle function in COVID-19 pathology are still scanty. Here, we performed a comparative study on exosomes recovered from the plasma of either MILD or SEVERE COVID-19 patients.
We show that although both types of vesicles efficiently display SARS-CoV-2 spike-derived peptides and carry immunomodulatory molecules, only those of MILD patients are capable of efficiently regulating antigen-specific CD4+ T-cell responses. Accordingly, by mass spectrometry, we show that the proteome of exosomes of MILD patients correlates with a proper functioning of the immune system, while that of SEVERE patients is associated with increased and chronic inflammation.
Overall, we show that exosomes recovered from the plasma of COVID-19 patients possess SARS-CoV-2-derived protein material, have an active role in enhancing the immune response, and possess a cargo that reflects the pathological state of patients in the acute phase of the disease.

Role of SARS-CoV-2 in causing blood-brain barrier leakage and microglial activation as a risk factor of cognitive deterioration in subjects at risk of Alzheimer’s disease

The recent pandemic provides evidence of altered central nervous system (CNS) function in response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-COV-2 invades the CNS by binding angiotensin-converting enzyme 2 (ACE2) expressed on neurons and glia. SARS-COV-2 may have effects on increased permeability of endothelial cells within the blood-brain barrier (BBB) as studies have shown that the S1 protein can transverse the BBB.
This is interesting because leakage of the BBB is implicated in Alzheimer’s disease (AD) pathogenesis. We hypothesized that SARS-CoV-2 infection leads to innate stimulated inflammation, ultimately activating microglial cells and an influx of pro-inflammatory cytokines and leukocytes in the meninges, contributing to increased permeability of the BBB. This BBB permeability increases AD susceptibility in subjects at risk by causing irreversible damage to the BBB and microglial cell activation.
Method: We developed a double transgenic mouse model using mice expressing human ACE2 receptor and 5xFAD mice that exhibit increased neuropathology seen in human AD allowing modeling of AD in SARS-CoV-2 pathogenesis. The hACE2/5xFAD double transgenic mice were intranasally inoculated with a sub-lethal dose of SARS-CoV-2 to test the hypothesis that SARS-COV-2 potentiates AD pathology and cognitive deterioration through impairment of the BBB. Leukocyte and cytokine populations were measured by flow cytometry and single-nuclei RNA sequencing of the meninges for characterization of microglial populations.
Result: SARS-CoV-2 creates a cytotoxic environment in the brain immediately following infection in hACE2/5xFAD mice leading to leakage of the BBB in the meninges. Activation of microglial innate cells by SARS-COV-2 invasion of the CNS will cause neural deterioration having long term implications on cognitive function. The hACE2/5xFAD mouse model allows us to uncover implications for SARS-COV-2 on AD cognitive deterioration.
Conclusion: The hACE2/5xFAD mouse allows modeling of SARS-CoV-2 in developing AD cases and allowed us to determine the immune environment generated in the meninges in response to SARS-CoV-2 infections. This mouse model provides a platform to proactively determine the effects of SARS-CoV-2 in developing AD cases, a methodology to be exploited for future mouse models determining the relationship of other viruses on AD pathology, and the opportunity to address phenotypes with therapeutics for preventative initiatives.
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Perspectives of farmers and tourists on agricultural abandonment in east Lesvos, Greece.

Perspectives of farmers and tourists on agricultural abandonment in east Lesvos, Greece.

Multi-stakeholder perceptions of panorama modifications are more and more acknowledged as important inputs to discussions on future panorama developments, significantly when addressing the way forward for European rural areas experiencing agricultural abandonment.

This analysis presents a case exploration of abandonment of olive plantations in east Lesvos, Greece. We carried out two units of semi-structured interviews to narrate an exploration on native farmers’ capacity and willingness to keep up the plantations, to the outcomes of a panorama choice survey undertaken with vacationers. Three farmer varieties are recognized following a cluster evaluation based mostly on attributes of particular person capacity and willingness to farm. Farmers belonging to the prevalent kind revealed low capacity and willingness and count on to additional extensify their farms.

The remaining two farmer varieties have greater willingness; they’re motivated by cultural causes, extra incessantly expressing a want to keep up their land underneath household possession, and partake in social cooperative initiatives selling practices valorizing the olive plantations. We define how these varieties work together with regional drivers of change, and partly additionally contribute to persistence of abandonment by way of constrained capacity to farm.

Abandonment doesn’t align with present panorama preferences of vacationers, who favor cultivated landscapes, components of traditionality inside constructed infrastructure and undertake nature-based actions. We focus on how excessive willingness to farm related to skilled and pluri-active types of farming could nevertheless present alternatives to keep up the cultivated panorama and synergize with (agri-)tourism demand. Our findings are corresponding to these of different European research, contributing to discussions on the way forward for its rural landscapes.

Perspectives of farmers and tourists on agricultural abandonment in east Lesvos, Greece.
Views of farmers and vacationers on agricultural abandonment in east Lesvos, Greece.

Immunophenotypic characterization, multi-lineage differentiation and growing older of zebrafish coronary heart and liver tissue-derived mesenchymal stem cells as a novel strategy in stem cell-based remedy.

Mesenchymal stem cells (MSCs) are a very good mannequin for preclinical and scientific investigations, and various sources of MSCs are topic to intensive experiments. On this examine, mesenchymal stem cells (MSCs) have been remoted from coronary heart and liver tissue of Zebrafish (Danio rerio). The flow-cytometry in addition to RT-PCR have been used to investigate the expression of a panel of cell floor markers CD44, CD90, CD31 and CD34.

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  1. Within the following, alizarin pink, oil red-O and toluidine blue staining have been carried out to judge the multi-lineage differentiation of zebrafish coronary heart and liver tissue-derived MSCs. Subsequently, the gene and protein expression of Oct4, Sox2 and Nanog as pluri

    -potent markers have been analyzed by RT-PCR and western blotting, respectively.

As well as, MTT assay was used for cell proliferation potential and inhabitants doubling time (PDT) evaluation. Additionally, the growing older of cells was investigated by β-galactosidase exercise assay. The outcomes confirmed that, like different MSCs, zebrafish coronary heart and liver tissue-derived MSCs have been constructive for mesenchymal, adverse for hematopoietic markers and expressed pluripotent markers Oct4, Sox2 and Nanog. Furthermore, these cells have been differentiated to osteocyte, adipocyte, and chondrocyte lineages following directed differentiation. It was discovered that PDT of zebrafish coronary heart and liver tissue-derived MSCs have been 50.67 and 46.61 h, respectively.

These cells had considerably extra speedy progress on day 4. Our outcomes present that zebrafish coronary heart and liver tissue-derived MSCs exhibited typical MSC traits together with fibroblast morphology, multi-lineage differentiation capability, pluriefficiency potential and expression of a typical set of basic MSC floor markers.

Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.

Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.

Platinum-based chemotherapy is the mainstay of front-line therapy of sufferers affected by pluri-metastatic intermediate/excessive grade NeuroEndocrine Neoplasms (NENs). Nevertheless, there aren’t any commonplace second-line remedies at illness development.

Earlier scientific experiences have evidenced that temozolomide (TMZ), an oral analog of dacarbazine, is lively in opposition to NENs at commonplace doses of 150 to 200 mg/mq per day on days 1 to five of a 28-day cycle, even when a major treatment-related toxicity is reported.

METHODS

Metastatic NENs sufferers had been handled on the ENETS (European NeuroEndocrine Tumor Society) heart of excellence of Naples (Italy), from 2014 to 2017 with a second-line various metronomic schedule of TMZ, 75 mg/m2per os “one week on/one week off”. Toxicity was graded with NCI-CTC standards v4.0; goal responses with RECIST v1.1 and efficiency standing (PS) in accordance with ECOG.

Twenty-six consecutive sufferers had been handled. Median age was 65.5 years. The predominant main organs had been pancreas and lung. Grading was G2 in 11 sufferers, G3 in 15. Greater than half of sufferers had a PS 2 (15 vs. 11 with PS 1). The median time-on-temozolomide remedy was 12.2 months (95% CI: 11.4-19.6). No G3/G4 toxicities had been registered.

Full response was obtained in 1 affected person, partial response in 4, steady illness in 19 (illness management price: 92.3%), and progressive illness in 2. The median total survival from TMZ begin was 28.Three months. PS improved in 73% of sufferers.Metronomic TMZ is an acceptable therapy for G2 and G3 NENs significantly in PS 2 sufferers. Potential and bigger trials are wanted to verify these outcomes.

Safety and Activity of Metronomic Temozolomide in Second-Line Treatment of Advanced Neuroendocrine Neoplasms.
Security and Exercise of Metronomic Temozolomide in Second-Line Remedy of Superior Neuroendocrine Neoplasms.

Would ivermectin for malaria management be useful in onchocerciasis-endemic areas?

There may be accumulating proof supporting using ivermectin as a malaria management software. Latest findings from the repeat ivermectin mass drug administrations for management of malaria trial demonstrated a diminished incidence of malaria in villages which obtained repeated ivermectin mass drug administration (MDA; six doses) in comparison with those that had just one spherical of ivermectin.

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A number of different research investigating the advantages of ivermectin for malaria functions are ongoing/deliberate.Whereas ivermectin MDA provides promising views within the combat in opposition to malaria, we spotlight the added advantages and anticipated challenges of conducting future research in onchocerciasis-endemic areas, that are confronted with a considerable illness burden together with onchocerciasis-associated epilepsy.

Rising the frequency of ivermectin MDA in such locations could scale back the burden of each malaria and onchocerciasis, and permit for extra entomological investigations on each the Anopheles mosquitoes and the blackflies.

Upfront, acceptability and feasibility research are wanted to evaluate the endorsement by the native populations, in addition to the programmatic feasibility of implementing ivermectin MDA a number of occasions a yr.Onchocerciasis-endemic websites would doubly profit from ivermectin MDA interventions, as these will alleviate onchocerciasis-associated morbidity and mortality, whereas doubtlessly curbing malaria transmission. Involving onchocerciasis applications and different related stakeholders within the malaria/ivermectin analysis agenda would foster the implementation of pluri-annual MDA in goal communities.

The ZT Biopolymer: A Self-Assembling Protein Scaffold for Stem Cell Applications.

The ZT Biopolymer: A Self-Assembling Protein Scaffold for Stem Cell Applications.

The event of cell tradition programs for the naturalistic propagation, self-renewal and differentiation of cells ex vivo is a excessive purpose of molecular engineering. Regardless of vital success in recent times, the excessive value of up-scaling cultures, the necessity for xeno-free tradition situations, and the diploma of mimicry of the natural extracellular matrix attainable in vitro utilizing designer substrates proceed to pose obstacles to the interpretation of cell-based technologies.

On this regard, the ZT biopolymer is a protein-based, steady, scalable, and economical cell substrate of excessive promise. ZT relies on the naturally occurring meeting of two human proteins: titin-Z1Z2 and telethonin. These protein constructing blocks are strong scaffolds that may be conveniently functionalized with full-length proteins and bioactive peptidic motifs by genetic manipulation, previous to self-assembly.

The polymer is, thereby, totally encodable. Functionalized variations of the ZT polymer have been proven to efficiently maintain the long-term culturing of human embryonic stem cells (hESCs), human induced pluripotent stem cells (hiPSCs), and murine mesenchymal stromal cells (mMSCs).

Pluripotency of hESCs and hiPSCs was retained for the longest interval assayed (four months). Outcomes level to the big potential of the ZT system for the creation of a modular, pluri-functional biomaterial for cell-based purposes.

The ZT Biopolymer: A Self-Assembling Protein Scaffold for Stem Cell Applications.

Maxwell®: An Unsupervised Studying Method for 5P Medication.

Within the 5P medication (Customized, Preventive, Participative, Predictive and Pluri-expert), the final pattern is to course of information by displacing the barycenter of the data from hospital centered programs to the affected person centered ones by his private medical information.

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In the present day, using synthetic intelligence for supporting this transition exhibits actual limitations in its implementation in operational follow, each on the stage of affected person care, but in addition within the basic each day lifetime of the well being skilled, due to the medico-legal imperatives induced by the guarantees of the ‘5P medication’.

On this paper, we suggest to fill this hole by introducing an authentic synthetic intelligence platform, named Maxwell, which follows an unsupervised studying method consistent with the medico-legal imperatives of the ‘5P medication’. We describe the useful platform traits and illustrate them by two examples of clustering in genomics and magnetic resonance imaging.