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1.
Biomedicines ; 12(4)2024 Apr 08.
Artículo en Inglés | MEDLINE | ID: mdl-38672177

RESUMEN

The entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into human embryonic kidney (HEK293T) cells has been shown to be a cholesterol-rich, lipid raft-dependent process. In this study, we investigated if the presence of a cholesterol uptake receptor Niemann-pick type c1-like1 (NPC1L1) impacts SARS-CoV-2 cell entry. Initially, we utilized reporter-based pseudovirus cell entry assays and a spike (S) glycoprotein-mediated cell-to-cell fusion assay. Using Chinese hamster ovary (CHO-K1) cells, which lack endogenous receptors for SARS-CoV-2 entry, our data showed that the co-expression of NPC1L1 together with the ACE2 receptor synergistically increased SARS-CoV-2 pseudovirus entry even more than the cells expressing ACE-2 receptor alone. Similar results were also found with the HEK293T cells endogenously expressing the ACE2 receptor. Co-cultures of effector cells expressing S glycoprotein together with target cells co-expressing ACE-2 receptor with NPC1L1 significantly promoted quantitative cell-to-cell fusion, including syncytia formation. Finally, we substantiated that an elevated expression of NPC1L1 enhanced entry, whereas the depletion of NPC1L1 resulted in a diminished SARS-CoV-2 entry in HEK293T-ACE2 cells using authentic SARS-CoV-2 virus in contrast to their respective control cells. Collectively, these findings underscore the pivotal role of NPC1L1 in facilitating the cellular entry of SARS-CoV-2. Importance: Niemann-Pick type C1-like1 (NPC1L1) is an endosomal membrane protein that regulates intracellular cholesterol trafficking. This protein has been demonstrated to play a crucial role in the life cycle of several clinically important viruses. Although SARS-CoV-2 exploits cholesterol-rich lipid rafts as part of its viral entry process, the role of NPC1L1 in SARS-CoV-2 entry remains unclear. Our research represents the first-ever demonstration of NPC1L1's involvement in facilitating SARS-CoV-2 entry. The observed role of NPC1L1 in human kidney cells is not only highly intriguing but also quite relevant. This relevance stems from the fact that NPC1L1 exhibits high expression levels in several organs, including the kidneys, and the fact that kidney damages are reported during severe cases of SARS-CoV-2. These findings may help us understand the new functions and mechanisms of NPC1L1 and could contribute to the identification of new antiviral targets.

2.
Daru ; 32(1): 461-468, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38613658

RESUMEN

PURPOSE: A pharmacopoeia is a compendium of guidelines and criteria for drug quality. It was established by a national or regional entity and has legal significance. This applies to administration of drugs in a particular nation or region. METHOD: In this study, the differences and similarities of microbiological acceptance criteria, specifications for microbial enumeration of herbal drugs and herbal drug preparations in 14 national and international pharmacopeias were investigated. RESULTS: It was found that 12 pharmacopeias have given separate microbial limits for total aerobic microbial count (TAMC) and total yeast and mold count (TYMC), and a list of specified microorganisms for which acceptance criteria are defined. However, similarities were noticed in Ph.Eur, Ph. Helv and, BP. Salmonella, and Escherichia coli are the most common pathogens specified for herbal preparations in which boiling water is added prior to use and for internal use in all Pharmacopoeias because they serve as indicators of potential contamination. CONCLUSION: From this study, it can be concluded that the differences in microbial limit tests and their acceptance criteria as specified in the various pharmacopoeias need to be harmonized. It will become a more convenient option for global drug manufacturers to import/export herbal drugs, and this would also eliminate the burden of performing various analytical methods and comply with different microbial acceptance criteria set by various pharmacopoeias. The comparative data obtained from this study will be used to develop strategies for revisions of pharmacopoeias in a harmonized manner with respect to microbiological acceptance criteria, specifications for microbial enumeration of herbal drugs and herbal drug preparations.


Asunto(s)
Contaminación de Medicamentos , Farmacopeas como Asunto , Preparaciones de Plantas , Preparaciones de Plantas/normas , Contaminación de Medicamentos/prevención & control , Farmacopeas como Asunto/normas , Recuento de Colonia Microbiana , Control de Calidad , Humanos
3.
Virus Res ; 345: 199375, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38642618

RESUMEN

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiologic agent of coronavirus disease 2019 (COVID-19), has posed significant challenges to global health. While much attention has been directed towards understanding the primary mechanisms of SARS-CoV-2 infection, emerging evidence suggests co-infections or superinfections with other viruses may contribute to increased morbidity and mortality, particularly in severe cases of COVID-19. Among viruses that have been reported in patients with SARS-CoV-2, seropositivity for Human cytomegalovirus (HCMV) is associated with increased COVID-19 risk and hospitalization. HCMV is a ubiquitous beta-herpesvirus with a seroprevalence of 60-90 % worldwide and one of the leading causes of mortality in immunocompromised individuals. The primary sites of latency for HCMV include CD14+ monocytes and CD34+ hematopoietic cells. In this study, we sought to investigate SARS-CoV-2 infection of CD14+ monocytes latently infected with HCMV. We demonstrate that CD14+ cells are susceptible and permissive to SARS-CoV-2 infection and detect subgenomic transcripts indicative of replication. To further investigate the molecular changes triggered by SARS-CoV-2 infection in HCMV-latent CD14+ monocytes, we conducted RNA sequencing coupled with bioinformatic differential gene analysis. The results revealed significant differences in cytokine-cytokine receptor interactions and inflammatory pathways in cells superinfected with replication-competent SARS-CoV-2 compared to the heat-inactivated and mock controls. Notably, there was a significant upregulation in transcripts associated with pro-inflammatory response factors and a decrease in anti-inflammatory factors. Taken together, these findings provide a basis for the heightened inflammatory response, offering potential avenues for targeted therapeutic interventions among HCMV-infected severe cases of COVID-19. SUMMARY: COVID-19 patients infected with secondary viruses have been associated with a higher prevalence of severe symptoms. Individuals seropositive for human cytomegalovirus (HCMV) infection are at an increased risk for severe COVID-19 disease and hospitalization. HCMV reactivation has been reported in severe COVID-19 cases with respiratory failure and could be the result of co-infection with SARS-CoV-2 and HCMV. In a cell culture model of superinfection, HCMV has previously been shown to increase infection of SARS-CoV-2 of epithelial cells by upregulating the human angiotensin-converting enzyme-2 (ACE2) receptor. In this study, we utilize CD14+ monocytes, a major cell type that harbors latent HCMV, to investigate co-infection of SARS-CoV-2 and HCMV. This study is a first step toward understanding the mechanism that may facilitate increased COVID-19 disease severity in patients infected with SARS-CoV-2 and HCMV.


Asunto(s)
COVID-19 , Infecciones por Citomegalovirus , Citomegalovirus , Receptores de Lipopolisacáridos , Monocitos , SARS-CoV-2 , Sobreinfección , Humanos , Monocitos/virología , Monocitos/inmunología , Citomegalovirus/inmunología , Receptores de Lipopolisacáridos/metabolismo , SARS-CoV-2/inmunología , COVID-19/virología , COVID-19/inmunología , Infecciones por Citomegalovirus/virología , Infecciones por Citomegalovirus/inmunología , Sobreinfección/virología , Sobreinfección/inmunología , Latencia del Virus , Inflamación , Coinfección/virología , Citocinas/metabolismo , Replicación Viral
4.
Trop Anim Health Prod ; 56(2): 49, 2024 Jan 18.
Artículo en Inglés | MEDLINE | ID: mdl-38236343

RESUMEN

The role of arthropods as livestock pests has been well established. Besides their biting habits causing nuisance in animals; they are important vectors for transmission of economically important livestock diseases worldwide. Various pests and vector control managemental programs that also make use of chemicals have variable success rates. Consequently, insecticide/acaricide resistance has been reported against most of the commonly used chemicals along with increased concern for environment and demand for clean and green, residue-free animal products. This calls for an urgent need to develop novel, alternate, effective strategies/technologies. This lays the foundation for the use of semiochemicals as alternatives along with other biological control agents. Current knowledge on semiochemical use in livestock is refined and limited; however, it has been widely exploited in the agricultural sector to control plant and food crop pests, surveillance, and monitoring. Semiochemicals have an added advantage of being natural and safe; however, knowledge of extraction and quantification by using assays needs to be explicit. Expertise is required in behavioral and electrophysiological studies of arthropods and their interactions with the host and environment targeting specific semiochemicals for promising results. A thorough prior understanding on aspects such as mechanism of action, the stimulus for the release, the effecter/target species, response produced, application methods, dose and concentration is required to develop any successful pest/vector control program. The current review provides essential and frontline information on semiochemicals and their potential applications in the livestock sector along with future challenges and interventions.


Asunto(s)
Acaricidas , Ganado , Animales , Agricultura , Feromonas , Control de Plagas
5.
Pathogens ; 13(1)2024 Jan 08.
Artículo en Inglés | MEDLINE | ID: mdl-38251367

RESUMEN

G-quadruplexes (G4s) are noncanonical nucleic acid structures that play significant roles in regulating various biological processes, including replication, transcription, translation, and recombination. Recent studies have identified G4s in the genomes of several viruses, such as herpes viruses, hepatitis viruses, and human coronaviruses. These structures are implicated in regulating viral transcription, replication, and virion production, influencing viral infectivity and pathogenesis. G4-stabilizing ligands, like TMPyP4, PhenDC3, and BRACO19, show potential antiviral properties by targeting and stabilizing G4 structures, inhibiting essential viral life-cycle processes. This review delves into the existing literature on G4's involvement in viral regulation, emphasizing specific G4-stabilizing ligands. While progress has been made in understanding how these ligands regulate viruses, further research is needed to elucidate the mechanisms through which G4s impact viral processes. More research is necessary to develop G4-stabilizing ligands as novel antiviral agents. The increasing body of literature underscores the importance of G4s in viral biology and the development of innovative therapeutic strategies against viral infections. Despite some ligands' known regulatory effects on viruses, a deeper comprehension of the multifaceted impact of G4s on viral processes is essential. This review advocates for intensified research to unravel the intricate relationship between G4s and viral processes, paving the way for novel antiviral treatments.

6.
ACS Omega ; 8(48): 45700-45707, 2023 Dec 05.
Artículo en Inglés | MEDLINE | ID: mdl-38075789

RESUMEN

This study showcases the creation of a biosensor strip designed for the rapid, precise, and highly sensitive electrochemical detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). These biosensor strips were crafted by affixing a monoclonal antibody (mAb) specific to SARS-CoV-2 onto the surface of a commercially screen-printed carbon electrode (SPCE) modified with polyaniline-titania nanotubes (PANi-TNT). The transportable sensing device was constructed by pairing the mAb functionalized strip with a portable potentiostat wirelessly connected to either a Windows or Android device. Fast and specific conjugation between spike protein of SARS-CoV-2 and immobilized anti-SARS-CoV-2 triggered a change in the charge and electron mobility in the biosensing layer of the strip to produce detectable current during chronoamperometric scanning in the presence of a phosphate buffer solution (PBS). The excellent sensitivity and specificity of the sensor toward SARS-CoV-2 were detected as analytical analysis demonstrated linearity in the range of 80 to 200 copies/µL with a limit of detection of 25.59 copies/µL from the dose-response and standard fitted curve. Through experimental validation, the sensor strip's ability to specifically detect SARS-CoV-2 was established, distinguishing it from human coronavirus-OC43 (HCoV-OC43), HCoV-NL63, HCoV-229E, and adenovirus. The results from these tests indicate that these strips possess the potential for the future creation of dependable and easily transportable point-of-care diagnostic devices, enabling swift, sensitive, and precise detection of SARS-CoV-2 in the saliva or nasopharyngeal fluid of individuals infected with the virus.

7.
Viruses ; 15(12)2023 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-38140679

RESUMEN

Kaposi's sarcoma-associated herpesvirus (KSHV) establishes life-long latent infection and is linked to several human malignancies. Latency-associated nuclear antigen (LANA) is highly expressed during latency, and is responsible for the replication and maintenance of the viral genome. The expression of LANA is regulated at transcriptional/translational levels through multiple mechanisms, including the secondary structures in the mRNA sequence. LANA mRNA has multiple G-quadruplexes (G4s) that are bound by multiple proteins to stabilize/destabilize these secondary structures for regulating LANA. In this manuscript, we demonstrate the role of Nucleolin (NCL) in regulating LANA expression through its interaction with G-quadruplexes of LANA mRNA. This interaction reduced LANA's protein expression through the sequestration of mRNA into the nucleus, demonstrated by the colocalization of G4-carrying mRNA with NCL. Furthermore, the downregulation of NCL, by way of a short hairpin, showed an increase in LANA translation following an alteration in the levels of LANA mRNA in the cytoplasm. Overall, the data presented in this manuscript showed that G-quadruplexes-mediated translational control could be regulated by NCL, which can be exploited for controlling KSHV latency.


Asunto(s)
G-Cuádruplex , Herpesvirus Humano 8 , Humanos , Herpesvirus Humano 8/fisiología , Nucleolina , ARN Mensajero/genética , ARN Mensajero/metabolismo , Antígenos Virales/genética , Latencia del Virus/genética
8.
Nat Prod Res ; : 1-7, 2023 Nov 16.
Artículo en Inglés | MEDLINE | ID: mdl-37971976

RESUMEN

To determine how vulnerable various pea genotypes are to leafminer infestation, a field experiment was conducted. On the basis of the presence of mines on five randomly selected leaflets from the upper, middle and lower parts of the plant, observations of larvae were made throughout the growing season. The total phenols were determined using the method described by Bray and Thorpe (1954, Analysis of phenolic compounds of interest in metabolism. Methods Biochem Anal. 52:1-27) and absorbance at 650 nm was measured using a spectrophotometer. There was a negative correlation between leafminer infestation and total phenol content. The UHF Pea-12 genotype, characterised by the lowest total phenol concentration (20.87 mg/100 g), exhibited the highest level of leaflet infestation (17.33%). Although UHF Pea-1 genotype had the lowest mean leaflet infestation (6.58%), it also had the highest phenol concentration (41.91 mg per 100 g). In context with this, the present study highlights the significance of host-plant resistance (HPR) in pest management.

9.
Microbiol Spectr ; 11(6): e0531622, 2023 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-37800915

RESUMEN

IMPORTANCE: Biological processes originating from the DNA and RNA can be regulated by the secondary structures present in the stretch of nucleic acids, and the G-quadruplexes are shown to regulate transcription, translation, and replication. In this study, we identified the presence of multiple G-quadruplex sites in the region (oriLyt) of Kaposi's sarcoma-associated herpesvirus (KSHV) DNA, which is essential for DNA replication during the lytic cycle. We demonstrated the roles of these G-quadruplexes through multiple biochemical and biophysical assays in controlling replication and efficient virus production. We demonstrated that KSHV achieves this by recruiting RecQ1 (helicase) at those G-quadruplex sites for efficient viral DNA replication. Analysis of the replicated DNA through nucleoside labeling and immunostaining showed a reduced initiation of DNA replication in cells with a pharmacologic stabilizer of G-quadruplexes. Overall, this study confirmed the role of the G-quadruplex in regulating viral DNA replication, which can be exploited for controlling viral DNA replication.


Asunto(s)
G-Cuádruplex , Herpesvirus Humano 8 , Herpesvirus Humano 8/genética , Replicación Viral/genética , Replicación del ADN , ADN Viral/genética , Regulación Viral de la Expresión Génica
10.
Bull Entomol Res ; 113(5): 598-603, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37475661

RESUMEN

Eupeodes corollae (F.) (Diptera: Syrphidae) is the most abundant syrphid fly which is distributed worldwide and is the sole predator of aphids. Therefore, the present study was conducted to evaluate the predation rate and functional response of E. corollae against the cabbage aphid, Brevicoryne brassicae (L.). The experiment was carried out under laboratory conditions at 25 ± 2°C with 60-70% relative humidity. The results revealed that age-specific net predation rate (qx) increased after the 4th day and a peak was recorded on the 10th day of pivotal age in the third larval instar. The stable host kill rate and finite host kill rate of E. corollae were 18.63 and 21.07, respectively, against the B. brassicae and predicted that a mean of 20.78 aphids was needed for E. corollae to produce one offspring. A negative linear coefficient (P < 0) indicated the type II functional response for all larval instars of E. corollae against the B. brassicae. At higher prey density, the prey consumption was significantly at par with second and third instar larvae of E. corollae as the prey consumption was increased with increasing the prey density, which then decreased after attaining the upper asymptote (76.40 and 81.40% consumption, respectively). The Roger's predator random equation for type II functional response was fitted to estimate attack rate (a) and handling time (Th). The maximum prey consumption was recorded for third instar of E. corollae with a higher attack rate (0.336 h-1) and lower handling time (0.514 h) against B. brassicae, followed by the second and first instar. Thus, it is concluded that the third larval instar of E. corollae was the voracious feeder and used as an efficient biocontrol agent in the IPM programme.


Asunto(s)
Áfidos , Dípteros , Animales , Áfidos/fisiología , Larva/fisiología , Conducta Predatoria
11.
Crit Rev Food Sci Nutr ; : 1-23, 2023 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-37218679

RESUMEN

Bovine milk peptides are the protein fragments with diverse bioactive properties having antioxidant, anticarcinogenic, other therapeutic and nutraceutical potentials. These peptides are formed in milk by enzymatic hydrolysis, gastrointestinal digestion and fermentation processes. They have significant health impact with high potency and low toxicity making them a suitable natural alternative for preventing and managing diseases. Antibiotic resistance has increased the quest for better peptide candidates with antimicrobial effects. This article presents a comprehensive review on well documented antimicrobial, immunological, opioid, and anti-hypertensive activities of bovine milk peptides. It also covers the usage of computational biology tools and databases for prediction and analysis of the food-derived bioactive peptides. In silico analysis of amino acid sequences of Bos taurus milk proteins have been predicted to generate peptides with dipeptidyl peptidase IV inhibitory and ACE inhibitory properties, making them favorable candidates for developing blood sugar lowering drugs and anti-hypertensives. In addition to the prediction of new bioactive peptides, application of bioinformatics tools to predict novel functions of already known peptides is also discussed. Overall, this review focuses on the reported as well as predicted biologically active peptide of casein and whey proteins of bovine milk that can be utilized to develop therapeutic agents.

12.
Infect Genet Evol ; 111: 105434, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37059256

RESUMEN

In early 2020, the emergence and spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the human population quickly developed into a global pandemic. SARS-CoV-2 is the etiological agent of coronavirus disease 2019 (COVID-19) which has a broad range of respiratory illnesses. As the virus circulates, it acquires nucleotide changes. These mutations are potentially due to the inherent differences in the selection pressures within the human population compared to the original zoonotic reservoir of SARS-CoV-2 and formerly naïve humans. The acquired mutations will most likely be neutral, but some may have implications for viral transmission, disease severity, and resistance to therapies or vaccines. This is a follow-up study from our early report (Hartley et al. J Genet Genomics. 01202021;48(1):40-51) which detected a rare variant (nsp12, RdRp P323F) circulating within Nevada in mid 2020 at high frequency. The primary goals of the current study were to determine the phylogenetic relationship of the SARS-CoV-2 genomes within Nevada and to determine if there are any unusual variants within Nevada compared to the current database of SARS-CoV-2 sequences. Whole genome sequencing and analysis of SARS-CoV-2 from 425 positively identified nasopharyngeal/nasal swab specimens were performed from October 2020 to August 2021 to determine any variants that could result in potential escape from current therapeutics. Our analysis focused on nucleotide mutations that generated amino acid variations in the viral Spike (S) protein, Receptor binding domain (RBD), and the RNA-dependent RNA-polymerase (RdRp) complex. The data indicate that SARS-CoV-2 sequences from Nevada did not contain any unusual variants that had not been previously reported. Additionally, we did not detect the previously identified the RdRp P323F variant in any of the samples. This suggests that the rare variant we detected before was only able to circulate because of the stay-at-home orders and semi-isolation experience during the early months of the pandemic. IMPORTANCE: SARS-COV-2 continues to circulate in the human population. In this study, SARS-CoV-2 positive nasopharyngeal/nasal swab samples were used for whole genome sequencing to determine the phylogenetic relationship of SARS-CoV-2 sequences within Nevada from October 2020 to August 2021. The resulting data is being added to a continually growing database of SARS-CoV-2 sequences that will be important for understanding the transmission and evolution of the virus as it spreads around the globe.


Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , SARS-CoV-2/metabolismo , COVID-19/epidemiología , Filogenia , Nevada , Estudios de Seguimiento , Mutación , ARN Polimerasa Dependiente del ARN/genética , Nucleótidos , ARN , Glicoproteína de la Espiga del Coronavirus/genética
13.
One Health ; 16: 100536, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-37041760

RESUMEN

Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral genome in wastewater has proven to be useful for tracking the trends of virus prevalence within the community. The surveillance also provides precise and early detection of any new and circulating variants, which aids in response to viral outbreaks. Site-specific monitoring of SARS-CoV-2 variants provides valuable information on the prevalence of new or emerging variants in the community. We sequenced the genomic RNA of viruses present in the wastewater samples and analyzed for the prevalence of SARS-CoV-2 variants as well as other respiratory viruses for a period of one year to account for seasonal variations. The samples were collected from the Reno-Sparks metropolitan area on a weekly basis between November 2021 to November 2022. Samples were analyzed to detect the levels of SARS-CoV-2 genomic copies and variants identification. This study confirmed that wastewater monitoring of SARS-CoV-2 variants can be used for community surveillance and early detection of circulating variants and supports wastewater-based epidemiology (WBE) as a complement to clinical respiratory virus testing as a healthcare response effort. Our study showed the persistence of the SARS-CoV-2 virus throughout the year compared to a seasonal presence of other respiratory viruses, implicating SARS-CoV-2's broad genetic diversity and strength to persist and infect susceptible hosts. Through secondary analysis, we further identified antimicrobial resistance (AMR) genes in the same wastewater samples and found WBE to be a feasible tool for community AMR detection and monitoring.

15.
J Mol Diagn ; 25(4): 191-196, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36754279

RESUMEN

The coronavirus disease 2019 (COVID-19) pandemic has provided a stage to illustrate that there is considerable value in obtaining rapid, whole-genome-based information about pathogens. This article describes the utility of a commercially available, automated severe acute respiratory syndrome associated coronavirus 2 (SARS-CoV-2) library preparation, genome sequencing, and a bioinformatics analysis pipeline to provide rapid, near-real-time SARS-CoV-2 variant description. This study evaluated the turnaround time, accuracy, and other quality-related parameters obtained from commercially available automated sequencing instrumentation, from analysis of continuous clinical samples obtained from January 1, 2021, to October 6, 2021. This analysis included a base-by-base assessment of sequencing accuracy at every position in the SARS-CoV-2 chromosome using two commercially available methods. Mean turnaround time, from the receipt of a specimen for SARS-CoV-2 testing to the availability of the results, with lineage assignment, was <3 days. Accuracy of sequencing by one method was 100%, although certain sites on the genome were found repeatedly to have been sequenced with varying degrees of read error rate.


Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Prueba de COVID-19 , Biología Computacional
16.
Mol Microbiol ; 119(4): 439-455, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36708073

RESUMEN

The histone-like protein HU plays a diverse role in bacterial physiology from the maintenance of chromosome structure to the regulation of gene transcription. HU binds DNA in a sequence-non-specific manner via two distinct binding modes: (i) random binding to any DNA through ionic bonds between surface-exposed lysine residues (K3, K18, and K83) and phosphate backbone (non-specific); (ii) preferential binding to contorted DNA of given structures containing a pair of kinks (structure-specific) through conserved proline residues (P63) that induce and/or stabilize the kinks. First, we show here that the P63-mediated structure-specific binding also requires the three lysine residues, which are needed for a non-specific binding. Second, we demonstrate that substituting P63 to alanine in HU had no impact on non-specific binding but caused differential transcription of diverse genes previously shown to be regulated by HU, such as those associated with the organonitrogen compound biosynthetic process, galactose metabolism, ribosome biogenesis, and cell adhesion. The structure-specific binding also helps create DNA supercoiling, which, in turn, may influence directly or indirectly the transcription of other genes. Our previous and current studies show that non-specific and structure-specific HU binding appear to have separate functions- nucleoid architecture and transcription regulation- which may be true in other DNA-binding proteins.


Asunto(s)
Proteínas Bacterianas , Histonas , Histonas/metabolismo , Proteínas Bacterianas/metabolismo , Lisina , Proteínas de Unión al ADN/metabolismo , ADN/metabolismo , ADN Bacteriano/metabolismo
17.
Viruses ; 14(12)2022 11 23.
Artículo en Inglés | MEDLINE | ID: mdl-36560613

RESUMEN

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus responsible for the COVID-19 pandemic. From the onset of the pandemic, rapid antigen tests have quickly proved themselves to be an accurate and accessible diagnostic platform. The initial (and still most commonly used antigen tests) for COVID-19 diagnosis were constructed using monoclonal antibodies (mAbs) specific to severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein (NP). These mAbs are able to bind SARS-CoV-2 NP due to high homology between the two viruses. However, since first being identified in 2019, SARS-CoV-2 has continuously mutated, and a multitude of variants have appeared. These mutations have an elevated risk of leading to possible diagnostic escape when using tests produced with SARS-CoV-derived mAbs. Here, we established a library of 18 mAbs specific to SARS-CoV-2 NP and used two of these mAbs (1CV7 and 1CV14) to generate a prototype antigen-detection lateral flow immunoassay (LFI). A side-by-side analysis of the 1CV7/1CV14 LFI and the commercially available BinaxNOWTM COVID-19 Antigen CARD was performed. Results indicated the 1CV7/1CV14 LFI outperformed the BinaxNOWTM test in the detection of BA.2, BA.2.12.1, and BA.5 Omicron sub-variants when testing remnant RT-PCR positive patient nasopharyngeal swabs diluted in viral transport media.


Asunto(s)
COVID-19 , Humanos , COVID-19/diagnóstico , SARS-CoV-2/genética , Prueba de COVID-19 , Pandemias , Sensibilidad y Especificidad , Inmunoensayo/métodos , Antígenos , Anticuerpos Monoclonales
18.
ACS Appl Mater Interfaces ; 14(45): 50463-50474, 2022 Nov 16.
Artículo en Inglés | MEDLINE | ID: mdl-36335476

RESUMEN

SARS-CoV-2 and its variants that continue to emerge have necessitated the implementation of effective disinfection strategies. Developing self-disinfecting surfaces can be a potential route for reducing fomite transmissions of infectious viruses. We show the effectiveness of TiO2 nanotubes (T_NTs) on photocatalytic inactivation of human coronavirus, HCoV-OC43, as well as SARS-CoV-2. T_NTs were synthesized by the anodization process, and their impact on photocatalytic inactivation was evaluated by the detection of residual viral genome copies (quantitative real-time quantitative reverse transcription polymerase chain reaction) and infectious viruses (infectivity assays). T_NTs with different structural morphologies, wall thicknesses, diameters, and lengths were prepared by varying the time and applied potential during anodization. The virucidal efficacy was tested under different UV-C exposure times to understand the photocatalytic reaction's kinetics. We showed that the T_NT presence boosts the inactivation process and demonstrated complete inactivation of SARS-CoV-2 as well as HCoV-OC43 within 30 s of UV-C illumination. The remarkable cyclic stability of these T_NTs was revealed through a reusability experiment. The spectroscopic and electrochemical analyses have been reported to correlate and quantify the effects of the physical features of T_NT with photoactivity. We anticipate that the proposed one-dimensional T_NT will be applicable for studying the surface inactivation of other coronaviruses including SARS-CoV-2 variants due to similarities in their genomic structure.


Asunto(s)
COVID-19 , Nanotubos , Humanos , SARS-CoV-2 , Nanotubos/química
19.
Sci Rep ; 12(1): 16141, 2022 09 27.
Artículo en Inglés | MEDLINE | ID: mdl-36167869

RESUMEN

Detection of SARS-CoV-2 viral load in wastewater has been highly informative in estimating the approximate number of infected individuals in the surrounding communities. Recent developments in wastewater monitoring to determine community prevalence of COVID-19 further extends into identifying SARS-CoV-2 variants, including those being monitored for having enhanced transmissibility. We sequenced genomic RNA derived from wastewater to determine the variants of coronaviruses circulating in the communities. Wastewater samples were collected from Truckee Meadows Water Reclamation Facility (TMWRF) from November 2020 to June 2021. SARS-CoV-2 variants resulting from wastewater were compared with the variants detected in infected individuals' clinical specimens (nasal/nasopharyngeal swabs) during the same period and found conclusively in agreement. Therefore, wastewater monitoring for SARS-CoV-2 variants in the community is a feasible strategy as a complementary tool to clinical specimen testing in the latter's absence.


Asunto(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , COVID-19/epidemiología , Humanos , ARN , ARN Viral/genética , SARS-CoV-2/genética , Aguas Residuales
20.
Med Microecol ; 14: 100059, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-35945946

RESUMEN

SARS-CoV-2 spread rapidly, causing millions of deaths across the globe. As a result, demand for medical supplies and personal protective equipment (PPE) surged and supplies dwindled. Separate entirely, hospital-acquired infections have become commonplace and challenging to treat. To explore the potential of novel sterilization techniques, this study evaluated the disinfection efficacy of Fathhome's ozone-based, dry-sanitizing device by dose and time response. Inactivation of human pathogens was tested on non-porous (plastic) surfaces. 95.42-100% inactivation was observed across all types of vegetative microorganisms and 27.36% inactivation of bacterial endospores tested, with no residual ozone detectable after completion. These results strongly support the hypothesis that Fathhome's commercial implementation of gas-based disinfection is suitable for rapid decontamination of a wide variety of pathogens on PPE and other industrially relevant materials.

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