Virus Penetration Research Articles

Autonomous Vehicle Data Protection : A Review of Security Threats, Challenges and Protective Mechanism

The swift advancement of autonomous vehicle (AV) technology has presented notable obstacles with data security and privacy. This analysis examines major security threats that jeopardize user privacy and vehicle control, including data breaches, virus penetration, cyber attacks, illegal access, V2V and V2I system hacking, GPS spoofing, and data leaks. It also looks at the challenges developers have when putting strong security measures in place, considering how complicated AV infrastructures may be and how cloud, IoT, and AI are integrated. The paper outlines a number of defence tactics, such as blockchain technology, machine learning for anomaly detection, encryption, and secure communication protocols. In order to improve AV cyber security, it ends with suggestions for further study in fields like edge computing and quantum-resistant cryptography.

In silico investigation of therapeutic potential of phytosteroids against SARS-CoV2 infection via inhibition of viral multiplication and attenuation of host cytokine storm

SARS-CoV2 infection overcomes host cell membrane barrier, followed by utilization of host cellular processes for viral multiplication. Simultaneously, it triggers a cytokine storm within and around infected cells and tissues. Anti-inflammatory agents that can potentially inhibit viral penetration and multiplication within the host cells may be ideal drug candidates against COVID-19. Dietary phytosteroids have significant anti-inflammatory potential. Hence, the present study intends to investigate anti-viral potential of three dietary phytosteroids, namely, brassinolide, 28-homocastasterone and 24-epibrassinolide against SARS-CoV2 proteins, S1 spike protein, nucleocapsid protein and main protease enzyme, in addition to host pro-inflammatory proteins, interleukin-1, tumour necrosis factor-α, cyclooxygenase-2 and prostaglandin synthase, as drug targets. Molecular docking studies by AutoDock version 4.0 was performed. Brassinolide, 28-homocastasterone and 24-epibrassinolide exhibits high docking score against all the seven proteins, as compared to hydroxychloroquine. Brassinolide and 28- homocastasterone has maximum binding affinity for pro-inflammatory proteins and SARS-CoV2 proteins.Dietary phytosteroids may potentially attenuate cytokine storm with simultaneous inhibition of host entry and multiplication of SARS-CoV2. In-vitro and in-vivo anti-viral studies of plant steroids may provide a clear path for the identification and development of novel drug candidates against COVID-19, that also provides evidence for the concept of reverse pharmacognosy.

Anti-Herpetic Activity of Killer Peptide (KP): An In Vitro Study.

Antimicrobial peptides represent a promising alternative to traditional drugs in relation to cost, toxicity, and, primarily, the growing problem of drug resistance. Here, we report on the activity against HSV-1 and HSV-2 of a previously described wide-spectrum synthetic decapeptide, Killer Peptide (KP). As determined by plaque reduction assays, treatment with KP at 100 μg/mL resulted in a reduction in the viral yield titer of 3.5 Logs for HSV-1 and 4.1 Logs for HSV-2. Further evaluation of KP antiviral activity focused on the early stages of the virus replicative cycle, including the determination of the residual infectivity of viral suspensions treated with KP. A direct effect of the peptide on viral particles impairing virus absorption and penetration was shown. The toxicity profile proved to be extremely good, with a selectivity index of 29.6 for HSV-1 and 156 for HSV-2. KP was also active against acyclovir (ACV)-resistant HSV isolates, while HSV subcultures in the presence of sub-inhibitory doses of KP did not lead to the emergence of resistant strains. Finally, the antiviral action of KP proved to be synergistic with that of ACV. Overall, these results demonstrate that KP could represent an interesting addition/alternative to acyclovir for antiviral treatment.

The effect of sodium deoxyribonucleate with iron complex on the expression of surface markers of MT-4 cells infected with human immunodeficiency virus type 1 (HIV-1) (Retroviridae: Primate lentivirus group)

The persistence of immune dysfunction during therapy has serious consequences for the health of HIV-infected people. Therefore, an important direction is the search for drugs that can reduce the inflammatory potential of the immune system and serve as an additional component of antiviral therapy. Aim ‒ to study the effect of the immunomodulatory drug Sodium deoxyribonucleate with iron complex (DNA-Na-Fe) on the expression of activation markers in MT-4 cells infected with HIV-1. Expression levels of CD4, CD28, CD38, CD62L and HLA-DR proteins on the plasma membrane were measured in cells. To assess viral activity, the p24 protein was quantified by ELISA. The two cell variants with different replicative activity were analyzed. Control cells, cells with DNA-Na-Fe, infected cells and infected cells with DNA-Na-Fe were tested. Based on the results obtained, it can be concluded that antiviral activity of the drug in MT-4 cells infected with HIV-1 is associated with immunomodulatory activity that enhances the expression of membrane proteins CD4, CD28, CD38 and CD62L. Diversity in the effect of DNA-Na-Fe on the studied surface proteins expression in two cell lines indicates that they depend on the characteristics of the combined molecular biological processes occurring in cells. And the increased effects observed in a system with changes in replicative activity assumes its active participation in virus replication at the stages of virus penetration and budding. Studies have shown that DNA-Na-Fe has antiviral and immunomodulatory activity.

CD14 gene polymorphism in COVID-19 convalescents: analysis of (C-159)T

Cluster of differentiation 14 (CD14) is a pattern recognition receptor for lipopolysaccharide of gram-negative flora and has a close relationship with toll-like receptor 4 (TLR4). The interaction between CD14 and TLR 4 leads to the synthesis of cytokines such as interleukin-1, interleukin-6 and tumor necrosis factor-á. The TLR4 receptor is associated with the penetration of the new coronavirus infection virus into the cell, additionally stimulating the expression of the angiotensin converting enzyme 2 molecule and suppressing apoptosis in infected cells. Work on the study of the (159C)T polymorphism of the CD14 gene in COVID-19 is represented by only a few publications describing observations only in the Western European region. The purpose of our study was to study the association between single nucleotide polymorphism (SNP) of the CD14 gene – (159C)T (rs2569190) and susceptibility to a new coronavirus infection in the population of the Republic of Crimea. The study included 158 patients (87 women (55%) and 71 (45%) men, average age 45.6±6.14 years) who had COVID-19. Verification of previous COVID-19 was based on anamnestic data and data from studies performed at the time of illness (PCR). The control group consisted of 85 respondents who had not suffered a new coronavirus infection. To analyze the (C-159)T polymorphism of the CD14 gene, allele-specific PCR with electrophoretic detection in a 3% agarose gel (NPF "Litekh", Russia) was used. To compare the frequencies of allele combinations, the ÷2 test and odds ratio (95% CI) were used. The distribution of CD14 mutations followed Hardy-Weinberg equilibrium (p 0.05). The results of the study showed that the distribution of CD14 gene genotypes did not differ significantly in all study groups. The dominant genotype was the heterozygous genotype CT of the (C-159)T polymorphism of the CD14 gene. The analysis showed that the presence of CT and TT SNPs was not associated with the risk of developing a new coronavirus infection (p 0.05). Conclusions. The CD14 SNP (C-159)T is not associated with a higher risk of COVID-19 infection. The distribution of occurrence of SNP variants in the group of recovered individuals did not differ significantly from that in the control group (p 0.05).

PET/CT Targeted Tissue Sampling Reveals Intravenously Administered HGN194 IgG1 Affects HIV Distribution after Rectal Exposure.

Neutralizing monoclonal antibodies hold great potential for prevention of human immunodeficiency virus (HIV) acquisition. IgG is the most abundant antibody in human serum, has a long half-life, and potent effector functions, making it a prime candidate for an HIV prevention therapeutic. We combined Positron Emission Tomography imaging and fluorescent microscopy of 64Cu-labeled, photoactivatable-green fluorescent protein HIV (PA-GFP-BaL) and fluorescently labeled HGN194 IgG1 to determine whether intravenously instilled IgG influences viral interaction with mucosal barriers and viral penetration in colorectal tissue 2 h after rectal viral challenge. Our results show that IgG1 did not alter the number of virions found throughout the colon or viral penetration into the epithelium of the rectum or descending colon. A minor increase in virions was observed in the transverse colon of IgG1 treated animals. Overall, the number of viral particles found in the mesenteric lymph nodes was low. However, IgG1 administration resulted in a significant reduction of virions found in mesenteric lymph nodes. Taken together, our results show that HGN194 IgG1 does not prevent virions from penetrating into the colorectal mucosa but may perturb HIV virion access to the lymphatic system.

Highly restrictive and directional penetration of the blood cerebral spinal fluid barrier by JCPyV.

The human polyomavirus JCPyV is an opportunistic pathogen that infects greater than 60% of the world's population. The virus establishes a persistent and asymptomatic infection in the urogenital system but can cause a fatal demyelinating disease in immunosuppressed or immunomodulated patients following invasion of the CNS. The mechanisms responsible for JCPyV invasion into CNS tissues are not known but direct invasion from the blood to the cerebral spinal fluid via the choroid plexus has been hypothesized. To study the potential of the choroid plexus as a site of neuroinvasion, we used an adult human choroid plexus epithelial cell line to model the blood-cerebrospinal fluid (B-CSF) barrier in a transwell system. We found that these cells formed a highly restrictive barrier to virus penetration either as free virus or as virus associated with extracellular vesicles (EVJC+). The restriction was not absolute and small amounts of virus or EVJC+ penetrated and were able to establish foci of infection in primary astrocytes. Disruption of the barrier with capsaicin did not increase virus or EVJC+ penetration leading us to hypothesize that virus and EVJC+ were highly cell-associated and crossed the barrier by an active process. An inhibitor of macropinocytosis increased virus penetration from the basolateral (blood side) to the apical side (CSF side). In contrast, inhibitors of clathrin and raft dependent transcytosis reduced virus transport from the basolateral to the apical side of the barrier. None of the drugs inhibited apical to basolateral transport suggesting directionality. Pretreatment with cyclosporin A, an inhibitor of P-gp, MRP2 and BCRP multidrug resistance transporters, restored viral penetration in cells treated with raft and clathrin dependent transcytosis inhibitors. Because choroid plexus epithelial cells are known to be susceptible to JCPyV infection both in vitro and in vivo we also examined the release of infectious virus from the barrier. We found that virus was preferentially released from the cells into the apical (CSF) chamber. These data show clearly that there are two mechanisms of penetration, direct transcytosis which is capable of seeding the CSF with small amounts of virus, and infection followed by directional release of infectious virions into the CSF compartment.

Investigating the antiviral activity of Erigeron annuus (L.) pers extract against RSV and examining its active components

Ethnopharmacological relevanceThe plants in the genus Erigeron are known to exhibit antiviral activities, including those against the respiratory syncytial virus (RSV). In traditional medicine Erigeron annuus (L.) Pers (EA) has been used in the treatment of pulmonary diseases and acute infectious hepatitis. Aim of this studyThe aim of this study is to determine the optimum extraction method to produce the most potent anti-RSV extract, elucidate its mode and mechanisms of antiviral activity in both in vitro and in vivo models, and identify the chemical structures of the bioactive compounds. Materials and methodsThe whole plant of EA was extracted with ethyl acetate, methanol, ethanol, water, aqueous methanol (60, 80% and 100%) and aqueous ethanol (50, 75% and 95%) using maceration, reflux, and ultrasound-assisted extraction methods. The antiviral activities of the extracts were determined in vitro. The in vitro antiviral activities of the extracts were determined using Hep-2 cells. Four in vitro experiments were performed to determine the mode of antiviral activity of the most active extract, ethyl acetate fraction (EAE) of Erigeron annuus whole plant extract prepared by refluxing with 50% ethanol, by examining its ability to inactivate the virus directly, inhibit viral adsorption and penetration, inhibit viral replication and preventive effect. The effect of temperature and duration of treatment on these modes of action was also determined. The antiviral activity of the EAE was also assessed in vivo in a mouse model. The lung index, viral load, and lung tissue histology were measured. qRT-PCR and ELISA studies were performed to determine the expression of key genes (TLR-3 and TLR-4) and proteins (IL-2, IFN-γ, and TNF-α) related to RSV infection. The most active antiviral compound was isolated using chromatography techniques, and its chemical structure was identified through electrospray triple quadrupole mass spectroscopy and nuclear magnetic resonance spectroscopy. ResultsThe EAE was the most active on RSV. In vitro experiments showed that the antiviral activity of EAE is via direct inactivation, inhibition of entry, and inhibition of the proliferation of the virus. In vivo experiments showed that the EAE effectively inhibited the proliferation of RSV in the lungs and alleviated the lung tissue lesions in RSV-infected mice. The antiviral activity of the EAE is mediated by downregulating the expression of TLR3 and TLR4 in the lung, upregulating the expression of IL-2 and IFN-γ, and downregulating the expression of TNF-α. Apigenin 7-O-methylglucuronide was found to be a major bioactive compound in EAE. ConclusionsThe results of this study confirmed the antiviral activity of EA by inactivating, inhibiting the entry, and inhibiting the proliferation of RSV. The activity is mediated by regulating the immunity and inflammatory mediators. Apigenin 7-O-methylglucuronide is the bioactive compound present in EA.

In Vitro Antiviral Activity of Kalanchoe daigremontiana Extract against Human Herpesvirus Type 1.

Plant polyphenols possess diverse bioactivities, including antiviral activity against a broad spectrum of viruses. Here, we investigated the virucidal properties of an Kalanchoe daigremontiana extract using an in vitro model of human herpesvirus type 1 (HHV-1) infection. Chromatographic analysis indicated that the extract of Kalanchoe daigremontiana is rich in various compounds, among which are polyphenols with virucidal activity confirmed in the literature. We found that Kalanchoe daigremontiana extract shows an ability to prevent HHV-1 infection by direct inhibition of the virus attachment, penetration, and blocking of infection when used in pretreatment or post-entry treatment. Our results indicate that Kalanchoe daigremontiana extract may be a good candidate drug against HHV-1, both as a substance to prevent infection and to treat an already ongoing infection. Our findings illustrate that Kalanchoe daigremontiana could be a potential new candidate for clinical consideration in the treatment of HHV-1 infection alone or in combination with other therapeutics.

Oral Hydroalcoholic Extract of Licorice for Management of Mild to Moderate Coronavirus Disease-19 Patients: A Double-Blind Randomized Clinical Trial

Background: Licorice is proposed as a treatment option for coronavirus disease-19 (COVID-19) by inhibiting the virus binding, penetration and replication; it also modulates various cellular signaling pathways. In this double-blind randomized-controlled trial, efficacy of Licorice hydroalcoholic extract oral formulation has been evaluated in patients with mild–moderate COVID-19in an outpatient clinic. Methods: A total of 40 mild to moderate outpatient Covid-19cases were randomly assigned into either Licorice or placebo groups (n=20 in each). Patients in Licorice group were given 700 mg capsules of hydroalcoholic extract of Licorice, trice a day for 2 weeks, while controls were given placebo capsules that were filled with Avicel (microcrystalline cellulose). Blood samples were collected from all participants before starting the medication and on days three, seven, and 14 for laboratory parameters including quantitative C-reactive protein (CRP), WBCs, lymphocytes, and neutrophils counts. The clinical symptoms were also recorded. Results: Our results showed that Licorice extract significantly decreased the frequency of cough on day seven (P=0.004) and the frequency of dyspnea on day three (P=0.02). No significant differences were found in the frequency of nausea, diarrhea, body pain, sore throat, runny nose, dizziness, lethargy, and loss of taste or smell between the two groups. Patients in Licorice group showed significantly higher oxygen saturations on days three, seven, and 14. Other laboratory parameters did not show any changes between the two groups. Conclusion: Licorice extract improved some clinical symptoms such as cough and dyspnea while only increased the oxygen saturation. Further studies with larger sample sizes at different stages of Covid-19are suggested.

Anticoronavirus activity of rhizome of Dryopteris crassirhizoma via multistage targeting of virus entry and viral proteases, Mpro and PLpro

Ethnopharmacological relevanceThe rhizome of Dryopteris crassirhizoma Nakai (Dryopteridaceae, RDC), a traditional East Asian herbal medicine, possesses a broad spectrum of medicinal properties, including anti-inflammatory, anticancer, antibacterial, and antiviral activities. Aim of the studyThis study investigates the 30% ethanolic extract of RDC's antiviral potential against human coronavirus OC43 (HCoV-OC43), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and its variants infections. Materials and methodsA 30% ethanolic extract of RDC or its components, filixic acid ABA (PubChem CID: 15081408) and dryocrassin ABBA (PubChem CID: 3082025) were treated with Human Coronavirus infection (HCoV-OC43, SARS-CoV-2 and its variants). The base peak chromatogram of RDC was evaluated using UPLC-Q/TOF Mass to identify the RDC, and the quantitative analysis of RDC compounds was performed using LC-MS/MS. A cytopathic effect (CPE) reduction assay, Western blot, immunofluorescence staining of viral protein expression, and qRT-PCR were performed to quantify the viral RNA copy numbers and determine the antiviral activity. The time-of-addition assay, the virus attachment, penetration, and virucidal assays, and SARS-CoV-2 Mpro and PLpro activity assay were used to elucidate the mode of action. ResultsRDC exhibited dose-dependent inhibition of HCoV-OC43–induced cytopathic effects, reducing viral RNA copy numbers and viral protein levels. Time-of-addition assays indicated that RDC targets the early stages of the HCoV-OC43 life cycle, inhibiting virion attachment and penetration with virucidal activity. Notably, filixic acid ABA and dryocrassin ABBA, constituents of RDC, reduced HCoV-OC43 viral RNA loads. Furthermore, RDC effectively blocked viral entry in pseudotyped lentivirus assays, involving spike proteins of SARS-CoV-2 Delta plus and South Africa variants, as well as control lentiviral particles expressing vesicular stomatitis virus glycoprotein G. Additionally, RDC demonstrated inhibition of SARS-CoV-2 infection and its variants by targeting viral proteases, namely main protease (Mpro) and papain-like protease (PLpro). ConclusionsThese findings underscore RDC's multistage approach to targeting viral infections by impeding virus entry and inhibiting viral protease activity. Therefore, RDC holds promise as a potent, broad-spectrum anticoronaviral therapeutic agent.

Clinical efficacy and safety of Xuanfei Touxie Fang as a complementary medicine in the treatment of SARS-CoV-2 Omicron variant: A retrospective study

IntroductionTo explore the clinical efficacy and safety of Xuanfei Touxie Fang (XFTXF) as a complementary medicine in the treatment of SARS-CoV-2 Omicron variant. MethodsA retrospective study of 317 patients with ordinary symptoms admitted to our hospital from December 2022 to February 2023 were divided into the observation group (n = 224) and the control group (n = 93). The observation group was treated with Xuanfei Touxie Fang combined with conventional treatment and the control group treated with conventional treatment alone. Recovery time of symptoms, length of hospital stay, clinical cure rate and recovery rate of chest CT manifestations were compared between the two groups. ResultsThe time to recover from fever (median: 1 vs 3 days) and the length of hospital stay (median: 7 vs 9 days) of the observation group after treatment were significantly shorter than the control group. The difference was statistically significant (P < 0.05). The clinical cure rate (68.3% vs 54.84 %) and the recovery rate of chest CT manifestations (87.95% vs 66.67 %) of the observation group after treatment were higher than the control group. The difference was statistically significant (P < 0.05). There was no adverse reactions during the treatment for two groups. The difference was no statistically significant (P > 0.05). DiscussionNumerous studies have demonstrated that Xuanfei Touxie Fang, which is frequently used in respiratory diseases, has anti-coronavirus effects by inhibiting viral replication, release, penetration, and RNA synthesis. Therefore, we treated COVID-19 with it and evaluated its safety and effectiveness. ConclusionsXuanfei Touxie Fang as a complementary medicine has a significant effect in the treatment of COVID-19, which can reduce the duration of fever and the length of hospital stay, improve the clinical cure rate and recovery rate of chest CT manifestations.

A Host Receptor Nanodisc-Based Biosensor Platform for the Detection of a Specific Virus and Its Variants.

The host receptor is a key element in the initial stage of the virus entry into the host. The use of this host receptor is valuable as a sensing element for selectively and sensitively detecting specific viruses. Also, viruses tend to escape neutralizing antibodies through viral mutation but still utilize the cell entry process using the same host receptors, so it would be a powerful detection tool even for the mutant viruses. The angiotensin-converting enzyme 2 (ACE2) receptor, which is the representative host receptor, performs an essential function in facilitating viral penetration by interacting with the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. In this study, we introduce a novel approach, where we fabricated a carbon nanotube field-effect transistor (CNT-FET) sensor and combined it with ACE2 receptor-embedded nanodisc (ND). ACE2 was produced using an E. coli expression system, purified, and integrated into the ND platform. ACE2 NDs showed robust functionality through interactions with a pseudotyped virus (PV) containing the spike protein, enabling sensitive detection of both SARS-CoV-2 and its genetic variations at 102 PFU/mL. The ACE ND-based sensor exhibited excellent selectivity by accurately differentiating SARS-CoV-2 wild-type and variants (Omicron, Delta) from other viruses (ZIKA and MERS-CoV). As a result of comparative analysis, ACE2 ND showed approximately 49% superior long-term functionality up to the second week compared to that of soluble ACE2. These findings highlight the high selectivity and sensitivity of host receptor-based sensors for detecting viral variants and provide a promising tool to prevent the spread of unknown viruses.

Abstract 588: Preclinical evaluation of novel chimeric vesiculovirus mediated expression of proteolytic enzymes targeting two major stromal components of pancreatic ductal adenocarcinoma

Abstract The vesiculovirus genus has recently garnered interest as an oncolytic virotherapy for pancreatic ductal adenocarcinoma (PDAC). A new synthetic chimeric vesiculovirus, VMG, demonstrated efficacy and safety against PDAC both in vitro and in vivo. The therapeutic effectiveness of virotherapies against PDAC is often hindered by immunosuppressive factors in the tumor environment, as well as the presence of hyaluronic acid (HA) and collagen (CN), which increase tumor stiffness and interstitial fluid pressure, leading to reduced drug/oncolytic virus and immune cell penetration into tumor. To address this, we engineered VMG viruses incorporating enzymes PH20, collagenase (CNase), and novel proteolytic enzyme SMC, the latter exhibits both PH20 and CNase-like activity. We characterized these modified viruses for their cytotoxicity and performance in vitro. Various PDAC cell lines displayed varied sensitivity to VMG vectors, yet they showed consistent oncolytic effects relative to the parent VMG-GFP virus. Our results revealed no compromise in viral replication or apoptosis-inducing ability after transgene insertion, which is evident as infectious viral progenies were produced to approximately similar level of magnitude of around 108 TCID50/ml in all the cell line tested. Additionally, the VMG-SMC and VMG-hPH20-CNase variants demonstrated notable enzyme activity. To enhance clinical relevance, we utilized an ex vivo live tumor platform, which indicated around 20% tumor cell death using VMG vectors compared to that of the control, signifying tumor viability and virus replicability 72 hours post-infection. Fluorescent imagery highlighted the viruses' ability to spread and express functional enzymes within tumors. In summary, VMG vectors expressing stroma-degrading proteolytic enzymes can effectively express PH20, CNase and SMC enzymes that degrade stromal barrier of HA and CN without interrupting the replicability, performance, and oncolytic characteristics of the VMG vectors. Future research should prioritize the safety of these vectors and their potential to augment neoantigen expression and immune cell infiltration in vivo, which are the hallmarks of antitumor efficacy. Citation Format: Khandoker Usran Ferdous, Mulu Z. Tesfay, Camila C. Simoes, Aleksandra Cios, Bolni Marius Nagalo. Preclinical evaluation of novel chimeric vesiculovirus mediated expression of proteolytic enzymes targeting two major stromal components of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 588.

Humoral Immune Response to SARS-CoV-2 Spike Protein Receptor-Binding Motif Linear Epitopes.

The worldwide spread of SARS-CoV-2 has led to a significant economic and social burden on a global scale. Even though the pandemic has concluded, apprehension remains regarding the emergence of highly transmissible variants capable of evading immunity induced by either vaccination or prior infection. The success of viral penetration is due to the specific amino acid residues of the receptor-binding motif (RBM) involved in viral attachment. This region interacts with the cellular receptor ACE2, triggering a neutralizing antibody (nAb) response. In this study, we evaluated serum immunogenicity from individuals who received either a single dose or a combination of different vaccines against the original SARS-CoV-2 strain and a mutated linear RBM. Despite a modest antibody response to wild-type SARS-CoV-2 RBM, the Omicron variants exhibit four mutations in the RBM (S477N, T478K, E484A, and F486V) that result in even lower antibody titers. The primary immune responses observed were directed toward IgA and IgG. While nAbs typically target the RBD, our investigation has unveiled reduced seroreactivity within the RBD's crucial subregion, the RBM. This deficiency may have implications for the generation of protective nAbs. An evaluation of S1WT and S2WT RBM peptides binding to nAbs using microscale thermophoresis revealed a higher affinity (35 nM) for the S2WT sequence (GSTPCNGVEGFNCYF), which includes the FNCY patch. Our findings suggest that the linear RBM of SARS-CoV-2 is not an immunodominant region in vaccinated individuals. Comprehending the intricate dynamics of the humoral response, its interplay with viral evolution, and host genetics is crucial for formulating effective vaccination strategies, targeting not only SARS-CoV-2 but also anticipating potential future coronaviruses.

SARS-CoV-2 presence in recreational seawater and evaluation of intestine permeability: experimental evidence of low impact on public health.

Coastal seawater pollution poses a public health risk due to the potential ingestion of contaminated water during recreational activities. Wastewater-based epidemiology has revealed the abundant presence of SARS-CoV-2 in seawater emitted from wastewater outlets. The objective of this research was to investigate the impact of seawater on SARS-CoV-2 infectivity to assess the safety of recreational activities in seawater. Wild SARS-CoV-2 was collected from oral swabs of COVID-19 affected patients and incubated for up to 90 min using the following solutions: (a) standard physiological solution (control), (b) reconstructed seawater (3.5% NaCl), and (c) authentic seawater (3.8%). Samples were then exposed to two different host systems: (a) Vero E6 cells expressing the ACE2 SARS-CoV-2 receptor and (b) 3D multi-tissue organoids reconstructing the human intestine. The presence of intracellular virus inside the host systems was determined using plaque assay, quantitative real-time PCR (qPCR), and transmission electron microscopy. Ultrastructural examination of Vero E6 cells revealed the presence of virus particles at the cell surface and in replicative compartments inside cells treated with seawater and/or reconstituted water only for samples incubated up to 2 min. After a 90-min incubation, the presence of the virus and its infectivity in Vero E6 cells was reduced by 90%. Ultrastructural analysis performed in 3D epi-intestinal tissue did not reveal intact viral particles or infection signs, despite the presence of viral nucleic acid detected by qPCR. Indeed, viral genes (Orf1ab and N) were found in the intestinal luminal epithelium but not in the enteric capillaries. These findings suggest that the intestinal tissue is not a preferential entry site for SARS-CoV-2 in the human body. Additionally, the presence of hypertonic saline solution did not increase the susceptibility of the intestinal epithelium to virus penetration; rather, it neutralized its infectivity. Our results indicate that engaging in recreational activities in a seawater environment does not pose a significant risk for COVID-19 infection, despite the possible presence of viral nucleic acid deriving from degraded and fragmented viruses.

Multi-computer malware detection systems with metamorphic functionality

The need to develop new systems for detecting and counteracting malware remains relevant. In addition to malware detection methods, the need to develop new systems for detecting and counteracting malware has become increasingly important. The use of various detection systems and the formation of a variable architecture in them significantly improves the effectiveness of detection, since both for attackers in computer attacks and for malware, understanding the system is significantly complicated. In addition, such systems may contain baits, traps, and, accordingly, modifiable operating environments to deceptively execute programs for research. This paper develops a conceptual model of multicomputer systems, which is designed to ensure the functioning of antivirus bait and traps to detect malware and computer attacks in corporate networks. The proposed approach is intended to prevent and counteract metamorphic virus penetration. This paper presents the conceptual model of multicomputer systems and introduces a defining characteristic responsible for the control of decisions and other defining characteristics of the system. Methods for detecting metamorphic viruses with the possibility of their implementation in the architecture of multi-computer systems with bait and traps are developed so that the system directly joins the detection procedure through its components and decides on the presence of metamorphic code in the executable file. An implementation of a multi-computer malware detection system with metamorphic functionality was developed to prove the feasibility of the proposed conceptual architecture model and the developed methods for detecting metamorphic viruses. An experiment on the functioning of a multi-computer malware detection system was set up, and experimental studies were conducted. The conducted experiments included metamorphic virus detection. In addition, an experiment on the effectiveness of detecting the metamorphic code of viruses was conducted. The efficiency of detecting metamorphic virus code using the developed multi-computer system was also investigated, and the presence of improved detection was established. The directions of further work are to extend the results of this work to new types of malware.

CXCR4 Is a Potential Target for Anti-HIV Gene Therapy.

The human immunodeficiency virus (HIV) epidemic is a global issue. The estimated number of people with HIV is 39,000,000 to date. Antiviral therapy is the primary approach to treat the infection. However, it does not allow for a complete elimination of the pathogen. The advances in modern gene therapy methods open up new possibilities of effective therapy. One of these areas of possibility is the development of technologies to prevent virus penetration into the cell. Currently, a number of technologies aimed at either the prevention of virus binding to the CCR5 coreceptor or its knockout are undergoing various stages of clinical trials. Since HIV can also utilize the CXCR4 coreceptor, technologies to modify this receptor are also required. Standard knockout of CXCR4 is impossible due to its physiological significance. This review presents an analysis of interactions between individual amino acids in CXCR4 and physiological ligands and HIV gp120. It also discusses potential targets for gene therapy approaches aimed at modifying the coreceptor.

Isolation of Membrane Bound Mucins from Human Bronchial Epithelial Cells.

Membrane-bound mucins constitute a large portion of the periciliary layer of lung epithelial surfaces, and thus play an important role in many aspects of innate defense. The biophysical and biochemical properties of the membrane-bound mucins have important implications for mucociliary clearance, viral penetration, and potential therapeutics delivered to the airway surface. Hence, isolating them and determining these properties is important in understanding airways disease and ultimately in developing treatments. Here, we describe a method using isopycnic centrifugation to enrich and isolate shed membrane-bound mucins from the washings of human bronchial epithelial cell cultures.

Hematopoietic stem cell transplantation in the treatment of HIV infection – comparison of “Berlin patient”, “London patient” and “Dusseldorf patient”

Introduction and aim. Human Immunodeficiency Virus-1 (HIV-1) remains one of the major issues in global public health. The standard therapy for HIV-1 positive patients includes using antiretroviral therapy (ART). These medications ensure suppression of viral replication but do not lead to a cure for the patient. The aim of this study was to present hematopoietic stem cell transplantation (HSCT) as a malignant treatment method which led to cure for three HIV-1 positive patients. Material and methods. Literature available in April 2023 was searched by using the PubMed and Google Scholar databases. Articles were selected using the following words: HIV, AIDS, HSCT therapy, ART therapy. Analysis of the literature. In each case of these described HSCT, the donor of hematopoietic stem cells had a homozygous mutation in the HIV co-receptor CCR5 (CCR5Δ32/Δ32). This mutation leads to a permanent lack of the protein and prevents penetration of virus by using this receptor. After transplantation, all of these 3 patients remained virus-free despite discontinuation of ART therapy. Conclusion. More research is needed to reduce the risk of using HSCT and perhaps in the future be able to use this therapy in all HIV-infected people.