Cytomegalovirus in Adenoma and Carcinoma Lesions: Detecting Mono-Infection and Co-Infection in Salivary Glands.

Salivary glands' neoplasms are hard to diagnose and present a complex etiology. However, several viruses have been detected in these neoplasms, such as HCMV, which can play a role in certain cancers through oncomodulation. The co-infections between HCMV with betaherpesviruses (HHV-6 and HHV-7) and polyomaviruses (JCV and BKV) has been investigated. The aim of the current study is to describe the frequency of HCMV and co-infections in patients presenting neoplastic and non-neoplastic lesions, including in the salivary gland. Multiplex quantitative polymerase chain reaction was used for betaherpesvirus and polyomavirus quantification purposes after DNA extraction. In total, 50.7% of the 67 analyzed samples were mucocele, 40.3% were adenoma pleomorphic, and 8.9% were mucoepidermoid carcinoma. Overall, 20.9% of samples presented triple-infections with HCMV/HHV-6/HHV-7, whereas 9.0% were co-infections with HCMV/HHV-6 and HCMV/HHV-7. The largest number of co-infections was detected in pleomorphic adenoma cases. All samples tested negative for polyomaviruses, such as BKV and JCV. It was possible to conclude that HCMV can be abundant in salivary gland lesions. A high viral load can be useful to help better understand the etiological role played by viruses in these lesions. A lack of JCV and BKV in the samples analyzed herein does not rule out the involvement of these viruses in one or more salivary gland lesion subtypes.

High silent prevalence of human herpesvirus 1 (HSV-1) infection affecting the indigenous reservation of the municipality of Dourados, Central-West Brazil.

BACKGROUND: The indigenous population located in the central region of Brazil, is the second largest in terms of population size in the country. The Indigenous Reserve of Dourados has risk factors that increase the vulnerability of the indigenous population to infectious diseases, especially Human alphaherpesvirus (HSV-1), a neglected disease with high prevalence in priority populations in developing countries. The virus can also cause many more severe diseases, including widespread neonatal infections, herpetic keratitis, and herpes encephalitis, which can be fatal if left untreated. We estimated the prevalence of anti-HSV-1 antibodies and correlated it with the demographic and behavioral characteristics of the Indigenous population of the Jaguapirú and Bororó villages (Dourados, Mato Grosso do Sul (MS), Brazil). METHODS: Our approach was cross-sectional. From March 2017 to November 2018. Using anti-HSV-1 (Gg1) IgM and anti-HSV-1 (gG1) IgG Euroimmun and the detection and quantification of HSV-1 viral load in plasma samples, through real-time PCR. The maps were constructed using QGIS and the statistical analyses using R Studio software. RESULTS: A total of 1138 individuals (> 18 years old) were enrolled. The prevalence of anti-HSV-1 IgM and IgG were 20% and 97.5%, respectively. The prevalence of anti-HSV-1 antibodies for IgG was higher in both sexes. Anti-HSV-1 IgM antibodies were present in 17.1%, 21.2%, 12.5%, and 22% of the participants with urinary problems, genital wounds, genital warts, and urethral discharge, respectively. Real-time PCR was used for confirmatory testing; HSV-1 DNA was detected in 25.6% (54/211) of anti-HSV1 IgM-positive samples. Viral loads ranged from 5.99E + 02 to 3.36E + 13. CONCLUSIONS: The seroprevalence of HSV-1 IgM and detection of HSV-1 DNA in the Indigenous population confirmed high silent prevalence. Furthermore, the seroprevalence of HSV-1 in the Indigenous population was higher than that reported in the general adult Brazilian population. Various socioeconomic factors, drug use, and health and sexual behaviors could contribute to the facilitation of HSV-1 transmission in the Indigenous population. Our results may help develop culturally appropriate intervention programs that eliminate health access barriers and improve the implementation of public health policies aimed at promoting information regarding the prevention, treatment, and control of HSV-1 infection in Brazilian Indigenous populations.

Can habits and behaviors predict colonization by community-associated MRSA in patients admitted to a Brazilian hospital?

This study aimed to identify factors associated with colonization by community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in adult patients admitted to a Brazilian hospital. This is a cross-sectional study, in which patients underwent a nasal swab and were asked about hygiene behavior, habits, and clinical history. Among the 702 patients, 180 (25.6%) had S. aureus and 21 (2.9%) MRSA. The factors associated with MRSA colonization were attending a gym (OR 4.71; 95% CI; 1.42 - 15.06), smoking habit in the last year (OR 2.37; 95% CI; 0.88 - 6.38), previous hospitalization (OR 2.18; CI 95%; 0.89 - 5.25), and shared personal hygiene items (OR 1.99; 95% CI; 0.71 - 5.55). At the time of admission, colonization by CA-MRSA isolates was higher than that found in the general population. This can be an important public health problem, already endemic in hospitals, whose factors such as those associated with habits (smoking cigarettes) and behaviors (team sports practice and activities in gyms) have been strongly highlighted. These findings may help developing infection control policies, allowing targeting patients on higher-risk populations for MRSA colonization.

Aptamer-Based Recognition of Breast Tumor Cells: A New Era for Breast Cancer Diagnosis.

Breast cancer is one of the leading causes of death among women worldwide and can be classified into four major distinct molecular subtypes based on the expression of specific receptors. Despite significant advances, the lack of biomarkers for detailed diagnosis and prognosis remains a major challenge in the field of oncology. This study aimed to identify short single-stranded oligonucleotides known as aptamers to improve breast cancer diagnosis. The Cell-SELEX technique was used to select aptamers specific to the MDA-MB-231 tumor cell line. After selection, five aptamers demonstrated specific recognition for tumor breast cell lines and no binding to non-tumor breast cells. Validation of aptamer specificity revealed recognition of primary and metastatic tumors of all subtypes. In particular, AptaB4 and AptaB5 showed greater recognition of primary tumors and metastatic tissue, respectively. Finally, a computational biology approach was used to identify potential aptamer targets, which indicated that CSKP could interact with AptaB4. These results suggest that aptamers are promising in breast cancer diagnosis and treatment due to their specificity and selectivity.

Could Herpesviridae be the cause of severe acute hepatitis of unknown origin in children?

INTRODUCTION: Severe acute hepatitis (SAH) is defined by a severe inflammation of hepatocytes in the liver parenchyma which can lead to an acute liver failure, a clinical condition with high mortality rate that can be triggered by several factors but is usually associated to hepatotropic viruses' infection. In 2022, cases of children with severe acute hepatitis of unknown origin hospitalized in Glasgow, Scotland, were reported. Possible causes of this condition include, but are not limited to, undiagnosed viral (and non-viral) infections, autoimmune hepatitis, drug and/or chemical toxicity, mitochondrial chain respiratory and metabolic disorders. AREAS COVERED: Herpesviruses can cause severe acute hepatitis, but little is known about the role and the mechanisms of herpesviruses as a causative agent of this type of hepatitis. We review the role of herpesviruses as causative agent of SAH in children and other possible mechanisms involved in this disease. EXPERT OPINION: Differential diagnosis for herpesvirus in SAH should be implemented in all settings. Alternative fluids, such as saliva and dried blood, could be used in the diagnosis to overwhelm the availability of biological specimens at sufficient volume. In the future, genetic studies could also be added to increase the knowledge about severe acute hepatitis in children.

Can habits and behaviors predict colonization by community-associated MRSA in patients admitted to a Brazilian hospital?

ABSTRACT This study aimed to identify factors associated with colonization by community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in adult patients admitted to a Brazilian hospital. This is a cross-sectional study, in which patients underwent a nasal swab and were asked about hygiene behavior, habits, and clinical history. Among the 702 patients, 180 (25.6%) had S. aureus and 21 (2.9%) MRSA. The factors associated with MRSA colonization were attending a gym (OR 4.71; 95% CI; 1.42 - 15.06), smoking habit in the last year (OR 2.37; 95% CI; 0.88 - 6.38), previous hospitalization (OR 2.18; CI 95%; 0.89 - 5.25), and shared personal hygiene items (OR 1.99; 95% CI; 0.71 - 5.55). At the time of admission, colonization by CA-MRSA isolates was higher than that found in the general population. This can be an important public health problem, already endemic in hospitals, whose factors such as those associated with habits (smoking cigarettes) and behaviors (team sports practice and activities in gyms) have been strongly highlighted. These findings may help developing infection control policies, allowing targeting patients on higher-risk populations for MRSA colonization.

Next Generation of Ovarian Cancer Detection Using Aptamers.

Ovarian cancer is among the seven most common types of cancer in women, being the most fatal gynecological tumor, due to the difficulty of detection in early stages. Aptamers are important tools to improve tumor diagnosis through the recognition of specific molecules produced by tumors. Here, aptamers and their potential targets in ovarian cancer cells were analyzed by in silico approaches. Specific aptamers were selected by the Cell-SELEX method using Caov-3 and OvCar-3 cells. The five most frequent aptamers obtained from the last round of selection were computationally modeled. The potential targets for those aptamers in cells were proposed by analyzing proteomic data available for the Caov-3 and OvCar-3 cell lines. Overexpressed proteins for each cell were characterized as to their three-dimensional model, cell location, and electrostatic potential. As a result, four specific aptamers for ovarian tumors were selected: AptaC2, AptaC4, AptaO1, and AptaO2. Potential targets were identified for each aptamer through Molecular Docking, and the best complexes were AptaC2-FXYD3, AptaC4-ALPP, AptaO1-TSPAN15, and AptaO2-TSPAN15. In addition, AptaC2 and AptaO1 could detect different stages and subtypes of ovarian cancer tissue samples. The application of this technology makes it possible to propose new molecular biomarkers for the differential diagnosis of epithelial ovarian cancer.

Genomic surveillance: a potential shortcut for effective Chagas disease management.

Chagas disease is an enduring public health issue in many Latin American countries, receiving insufficient investment in research and development. Strategies for disease control and management currently lack efficient pharmaceuticals, commercial diagnostic kits with improved sensitivity, and vaccines. Genetic heterogeneity of Trypanosoma cruzi is a key aspect for novel drug design since pharmacological technologies rely on the degree of conservation of parasite target proteins. Therefore, there is a need to expand the knowledge regarding parasite genetics which, if fulfilled, could leverage Chagas disease research and development, and improve disease control strategies. The growing capacity of whole-genome sequencing technology and its adoption as disease surveillance routine may be key for solving this long-lasting problem.

Differences in Charge Distribution in Leishmania tarentolae Leishmanolysin Result in a Reduced Enzymatic Activity.

Leishmania tarentolae is a non-pathogenic trypanosomatid isolated from lizards widely used for heterologous protein expression and extensively studied to understand the pathogenic mechanisms of leishmaniasis. The repertoire of leishmanolysin genes was reported to be expanded in L. tarentolae genome, but no proteolytic activity was detected. Here, we analyzed L. tarentolae leishmanolysin proteins from the genome to the structural levels and evaluated the enzymatic activity of the wild-type and overexpressing mutants of leishmanolysin. A total of 61 leishmanolysin sequences were retrieved from the L. tarentolae genome. Five of them were selected for phylogenetic analysis, and for three of them, we built 3D models based on the crystallographic structure of L. major ortholog. Molecular dynamics simulations of these models disclosed a less negative electrostatic potential compared to the template. Subsequently, L. major LmjF.10.0460 and L. tarentolae LtaP10.0650 leishmanolysins were cloned in a pLEXSY expression system into L. tarentolae. Proteins from the wild-type and the overexpressing parasites were submitted to enzymatic analysis. Our results revealed that L. tarentolae leishmanolysins harbor a weak enzymatic activity about three times less abundant than L. major leishmanolysin. Our findings strongly suggest that the less negative electrostatic potential of L. tarentolae leishmanolysin can be the reason for the reduced proteolytic activity detected in this parasite.

Bacterial 2'-Deoxyguanosine Riboswitch Classes as Potential Targets for Antibiotics: A Structure and Dynamics Study.

The spread of antibiotic-resistant bacteria represents a substantial health threat. Current antibiotics act on a few metabolic pathways, facilitating resistance. Consequently, novel regulatory inhibition mechanisms are necessary. Riboswitches represent promising targets for antibacterial drugs. Purine riboswitches are interesting, since they play essential roles in the genetic regulation of bacterial metabolism. Among these, class I (2'-dG-I) and class II (2'-dG-II) are two different 2'-deoxyguanosine (2'-dG) riboswitches involved in the control of deoxyguanosine metabolism. However, high affinity for nucleosides involves local or distal modifications around the ligand-binding pocket, depending on the class. Therefore, it is crucial to understand these riboswitches' recognition mechanisms as antibiotic targets. In this work, we used a combination of computational biophysics approaches to investigate the structure, dynamics, and energy landscape of both 2'-dG classes bound to the nucleoside ligands, 2'-deoxyguanosine, and riboguanosine. Our results suggest that the stability and increased interactions in the three-way junction of 2'-dG riboswitches were associated with a higher nucleoside ligand affinity. Also, structural changes in the 2'-dG-II aptamers enable enhanced intramolecular communication. Overall, the 2'-dG-II riboswitch might be a promising drug design target due to its ability to recognize both cognate and noncognate ligands.

In silico investigation of riboswitches in fungi: structural and dynamical insights into TPP riboswitches in Aspergillus oryzae.

Riboswitches are RNA sensors affecting post-transcriptional processes through their ability to bind to small molecules. Thiamine pyrophosphate (TPP) riboswitch plays a crucial role in regulating genes involved in synthesizing or transporting thiamine and phosphorylated derivatives in bacteria, archaea, plants, and fungi. Although TPP riboswitch is reasonably well known in bacteria, there is a gap in the knowledge of the fungal TPP riboswitches structure and dynamics, involving mainly sequence variation and TPP interaction with the aptamers. On the other hand, the increase of fungal infections and antifungal resistance raises the need for new antifungal therapies. In this work, we used computational approaches to build three-dimensional models for the three TPP riboswitches identified in Aspergillus oryzae, in which we studied their structure, dynamics, and binding free energy change (ΔGbind) with TPP. Interaction patterns between the TPP and the surrounding nucleotides were conserved among the three models, evidencing high structural conservation. Furthermore, we show that the TPP riboswitch from the A. oryzae NMT1 gene behaves similarly to the E. coli thiA gene concerning the ΔGbind. In contrast, mutations in the fungal TPP riboswitches from THI4 and the nucleoside transporter genes led to structural differences, affecting the binding-site volume, hydrogen bond occupancy, and ΔGbind. Besides, the number of water molecules surrounding TPP influenced the ΔGbind considerably. Notably, our ΔGbind estimation agreed with previous experimental data, reinforcing the relationship between sequence conservation and TPP interaction.

Genomic surveillance: a potential shortcut for effective Chagas disease management

Chagas disease is an enduring public health issue in many Latin American countries, receiving insufficient investment in research and development. Strategies for disease control and management currently lack efficient pharmaceuticals, commercial diagnostic kits with improved sensitivity, and vaccines. Genetic heterogeneity of Trypanosoma cruzi is a key aspect for novel drug design since pharmacological technologies rely on the degree of conservation of parasite target proteins. Therefore, there is a need to expand the knowledge regarding parasite genetics which, if fulfilled, could leverage Chagas disease research and development, and improve disease control strategies. The growing capacity of whole-genome sequencing technology and its adoption as disease surveillance routine may be key for solving this long-lasting problem.

Agentes comunitários de saúde e o trabalho com usuários de drogas: obstáculos e desfechos possíveis / Community health agents and work with drug users: possible obstacles and outcomes / Agentes de salud comunitaria y el trabajo con usuarios de drogas: posibles obstáculos y resultados

INTRODUÇÃO: Na Estratégia Saúde da Família (ESF), o trabalho do Agente Comunitário de Saúde (ACS) é ponto inicial de ancoragem das ações em saúde voltadas à comunidade. OBJETIVO: compreender o entendimento dos ACS em relação à abordagem da temática das drogas no território em que atuam. Métodos: Estudo qualitativo, realizado com 6 ACS de uma Equipe de Unidade de ESF. Definiu-se a amostra por exaustão. Foram realizadas entrevistas, posteriormente transcritas e organizadas conforme o Discurso do Sujeito Coletivo. RESULTADOS: Foram evidenciados entraves e equívocos no processo de trabalho dos ACS no manejo das demandas relacionadas ao uso de álcool e drogas. CONCLUSÃO: A percepção dos ACS sobre a temática do uso de drogas no cotidiano do serviço de saúde é cautelosa e restrita. Mesmo diante da identificação disto como um problema do território, suas intervenções ocorrem mediante manifestação voluntária do usuário, quando este busca o serviço de saúde.

INTRODUCTION: In the Family Health Strategy (FHS), the work of the Community Health Agent (CHA) is the starting point for anchoring health actions aimed at the community. OBJECTIVE: to comprehend the understanding of the CHA in relation to addressing the theme of drugs in the territory in which they operate. Methods: Qualitative study, carried out with 6 CHA from an FHS Unit Team. The sample was defined by exhaustion. Interviews were carried out, later transcribed and organized according to the Collective Subject Discourse. RESULTS: Obstacles and misunderstandings were evidenced in the work process of the CHA in handling the demands related to the use of alcohol and drugs. CONCLUSION: The perception of CHAs on the theme of drug use in the daily routine of the health service is cautious and restricted. Even with the identification of this as a territory problem, their interventions occur through voluntary manifestation of the user, once he seeks the health service.

INTRODUCCIÓN: En la Estrategia de Salud Familiar (ESF), el trabajo del Agente de Salud Comunitaria (ASC) es el punto de partida para anclar las acciones de salud dirigidas a la comunidad. OBJETIVO: comprender lo entendimiento de la ASC en relación con el tema de las drogas en el territorio en el que operan. Métodos: Estudio cualitativo, realizado con 6 ASC de un equipo de la Unidad ESF. La muestra se definió por agotamiento. Las entrevistas se llevaron a cabo, luego se transcribieron y se organizaron de acuerdo con el Discurso del Sujeto Colectivo. RESULTADOS: se evidenciaron obstáculos y malentendidos en el proceso de trabajo de la ASC en el manejo de las demandas relacionadas con el uso de alcohol y drogas. CONCLUSIÓN: La percepción de los ASC sobre el tema del consumo de drogas en la vida diaria del servicio de salud es cautelosa y restringida. Incluso con la identificación de esto como un problema del territorio, sus intervenciones ocurren a través de la manifestación voluntaria del usuario, cuando este busca el servicio de salud.

Whole-Genome Sequences of Mycobacterium abscessus subsp. massiliense Isolates from Brazil.

The Mycobacterium abscessus complex comprises multidrug-resistant, opportunistic, and rapidly growing pathogens responsible for severe infections. Here, we report the genome composition of four Mycobacterium abscessus subsp. massiliense isolates from three sources: two from the lung of a cystic fibrosis patient, one from a mammary cyst, and one from a gutter system.

Using 14C-glyphosate to investigate the distribution of two formulations in transgenic glyphosate-resistant soybean.

Glyphosate residues in grain can be explained by the concentrations and formulations of glyphosate products. This study aimed to evaluate the residues from glyphosate formulations labeled with 14C-glyphosate applied to leaves of glyphosate-resistant soybean (GRS) in two life cycles by liquid scintillation spectrometry. Different plant tissues were analyzed after the end of the plants' life cycles. The experimental design was four repetitions of three treatments: Roundup® Original, Roundup Ready®, and unformulated glyphosate (control). The application of the dosing solution was 120 µL on the first four trifoliate leaves (10 µL per leaflet) of each plant, deposited manually with a 1-µL dispenser. All treatment solutions were calculated at a 1.2 kg a.e ha-1 of glyphosate. Glyphosate formulations of Roundup® Original and Roundup Ready® increased 14C-glyphosate distribution in GRS compared to the unformulated herbicide, regardless of the experiment (first or second cycle). Overall, the percentages of total radioactivity applied (18.33 kBq) found in grains were less than 5%. Grains, stems, and leaves showed the highest levels of herbicide residues compared to other parts of the plant. Despite the Roundup Ready® formulation having increased residues, the highest value found in grains, 1.95 mg kg-1, was less than 10 mg kg-1, the maximum residue limit (MRL) in Brazil.

Plasmodium vivax metacaspase 1 (PvMCA1) catalytic domain is conserved in field isolates from Brazilian Amazon.

In the present study, we investigated the genetic diversity of Plasmodium vivax metacaspase 1 (PvMCA1) catalytic domain in two municipalities of the main malaria hotspot in Brazil, i.e., the Juruá Valley, and observed complete sequence identity among all P. vivax field isolates and the Sal-1 reference strain. Analysis of PvMCA1 catalytic domain in different P. vivax genomic sequences publicly available also revealed a high degree of conservation worldwide, with very few amino acid substitutions that were not related to putative histidine and cysteine catalytic residues, whose involvement with the active site of protease was herein predicted by molecular modeling. The genetic conservation presented by PvMCA1 may contribute to its eligibility as a druggable target candidate in vivax malaria.

M. bovis BCG Moreau N-Terminal Loss Leads to a Less Stable Dodecin With Lower Flavin Binding Capacity.

Tuberculosis still remains a concerning health problem worldwide. Its etiologic agent, Mycobacterium tuberculosis, continues to be the focus of research to unravel new prophylactic and therapeutic strategies against this disease. The only vaccine in use against tuberculosis is based on the in vitro attenuated strain, M. bovis BCG. Dodecin is a dodecameric complex important for flavin homeostasis in Archea and Eubacteria, and the M. tuberculosis protein is described as thermo- and halostable. M. bovis BCG Moreau, the Brazilian vaccine strain, has a single nucleotide polymorphism in the dodecin start codon, leading to a predicted loss of seven amino acids at the protein N-terminal end. In this work we aimed to characterize the effect of this mutation in the BCG Moreau protein features. Our recombinant protein assays show that the predicted BCG homolog is less thermostable than M.tb's but maintains its dodecamerization ability, although with a lower riboflavin-binding capacity. These data are corroborated by structural analysis after comparative modeling, showing that the predicted BCG dodecin complex has a lower interaction energy among its monomers and also a distinct electrostatic surface near the flavin binding pocket. However, western blotting assays with the native proteins were unable to detect significant differences between the BCG Moreau and M.tb orthologs, indicating that other factors may be modulating protein structure/function in the bacterial context.

Deteção de citomegalovirus, herpesvirus humano 6 e herpesvirus humano 7 em amostras de lesões em glândula salivar parafinadas / Detection of cytomegalovirus, human herpesvirus 6 and human herpesvirus 7 in paraffin salivary gland lesion samples

Guimarães Os vírus têm sido considerados como importantes patógenos no desenvolvimento de neoplasias em glândulas salivares, dentre estes, destacam-se os Betaherpesvirus humano como o Citomegalovirus humano (HCMV), Herpesvirus humano 6 (HHV-6) e Herpesvirus humano 7 (HHV-7). Os betaherpesvírus são característicos por apresentarem alta prevalência na população mundial, sem apresentar sazonalidade, capazes de causar infecção latente e reativação viral em seus hospedeiros. Devido a detecção destes vírus em amostras de saliva, tem sido proposto em alguns estudos a ação de betaherpesvírus em algumas patogenias de glândulas salivares, como a formação de patogenias nestes órgãos. Neoplasias em glândulas salivares representam cerca de 3-6% de todas as neoplasias de cabeça e pescoço, com incidência mundial anual de aproximadamente 0,4-13,5% por 100.000 indivíduos. Apesar de haver dados de detecção de betaherpesvírus em neoplasias salivares e em amostras de saliva, ainda existem estudos insuficientes que explorem o papel destes vírus nestas patogenias salivares. O objetivo deste estudo foi investigar a presença dos Betaherpesvirus humano (HCMV, HHV-6 e HHV-7) em neoplasias de glândula salivar parafinadas. Um ensaio de qPCR foi realizado para amplificação das regiões U54, U56 e U37 do HCMV, HHV-6 e HHV-7, respectivamente para quantificar a carga viral em 68 amostras parafinadas de lesões salivares. Dentre as 68 amostras processadas, 51,4% eram de mucocele (35/68), 39,7% de adenoma pleomórfico (27/68) e 8,8% de carcinoma mucoepidermoóide (6/68). A detecção de betaherpesvírus nestas lesões foi alta, apresentando maior detecção para HCMV com 52,9%, 47,05% para HHV-6 e 39,7% para HHV-7, possuindo predominância de detecção de betaherpesvírus na lesão do tipo adenoma pleomórfico. Foi observado que 50,0% das amostras apresentaram tripla-infecção por HCMV/HHV-6/HHV-7, sendo detectado 20,0% de coinfecções por HCMV/HHV-6, 20,0% de HCMV/HHV-7 e 10,0% de HHV-6/HHV-7, com coinfecções ocorrendo na lesão do tipo adenoma pleomórfico em maior taxa. A alta detecção de HCMV, HHV-6 e HHV-7 em glândulas salivares, indica que este órgão pode ser possível de sítio de replicação destes vírus

The viruses have been considered as important pathogens in the development of neoplasms in salivary glands, between these, Human betaherpesvirus such as Human cytomegalovirus (HCMV), Human herpesvirus 6 (HHV-6) and Human herpesvirus 7 (HHV-7) stand out. Betaherpesvirus is characteristic because they have a high prevalence in the world population, without presenting seasonality, capable of causing latent infection and viral reactivation in their hosts. Due to the detection of these viruses in saliva samples, the action of betaherpesvirus in some pathogenesis of salivary glands, such as the formation of pathogenesis in these organs, has been proposed in some studies. Salivary gland neoplasms account for about 3-6% of all head and neck neoplasms, with an annual worldwide incidence of approximately 0,4-13,5% per 100,000 individuals. Although there are data for the detection of betaherpesvirus in salivary neoplasms and saliva samples, thereare still insufficient studies exploring the role of these viruses in these salivary pathogeneses. The aim of this study was to investigate the presence of human Betaherpesvirus (HCMV, HHV-6 and HHV-7) in paraffin salivary gland neoplasms. A qPCR assay was performed to amplify the U54, U56 and U37 regions of HCMV, HHV-6 and HHV-7, respectively to quantify the viral load in 68 paraffin samples of salivary lesions. Among the 68 samples processed,51,4%were mucocele (35/68), 39.7% pleomorphic adenoma (27/68) and 8,8% mucoepidermoid carcinoma (6/68). The detection of betaherpesvirus in these lesions was high, presenting higher detection for HCMV with 52,9%, 47,05% for HHV6 and 39,7% for HHV-7, with predominance of detection of betaherpesvirus in the lesion of the pleomorphic adenoma type. It was observed that 50.0% of the samples presented triple-infection by HCMV/HHV-6/HHV-7, and 20.0% of co-infections by HCMV/HHV-6, 20.0% of HCMV/HHV-7 and 10.0% of HHV-6/HHV-7 were detected, with co-infections occurring with higher predominance of pleomorphic adenoma lesion. The high detection of HCMV, HHV-6 and HHV-7 in salivary glands indicates that this organ may be possible from replication site.

Shared Binding Mode of Perrottetinene and Tetrahydrocannabinol Diastereomers inside the CB1 Receptor May Incentivize Novel Medicinal Drug Design: Findings from an in Silico Assay.

In recent years, therapeutic compounds derived from phytocannabinoids have brought renewed attention to the benefits they offer to ameliorate chronic disease symptoms. Among cannabinoids, tetrahydrocannabinol (THC) is a well-known component of the Cannabis plant, whose active principles have been studied through the years. Another psychoactive phytocannabinoid, derived from liverworts Radula, perrottetinene (PET), has created interest, especially as a pharmaceutical product and for its legal recreational use. Unfortunately, so far, the interaction mode of these compounds at the type 1 cannabinoid receptors (CB1R) binding site remains unknown, and no experimental three-dimensional structure in complex with THC or PET is available in the Protein Data Bank. Today, many computational methodologies can assist in this crusade and help unveil how these molecules bind, based on the already known pose of a structurally similar compound. In this work, we aim to elucidate the binding mode of THC and PET molecules in both cis and trans conformers, using a combination of several computational methodologies, including molecular docking, molecular dynamics, free energy calculations, and protein-energy network studies. We found that THC and PET interact similarly with the CB1R, in a different conformation depending on the considered diastereomer. We have observed that cis ligands adopted a half-chair conformation of the cycle ring containing the dimethyl group, assuming an axial or equatorial conformation producing a different induced fitting of the surrounding residues compared with trans ligands, with higher interaction energy than the trans conformer. For PET, we have seen that Trp-279 and Trp-356 have a marked influence on the binding. After binding, Trp-279 accommodates its side chain to better interact with the PET's terminal phenyl group, disturbing CB1R residues communication. The interaction with Trp-356 might impair the activation of CB1R and can influence the binding of PET as a partial agonist. Understanding the PET association with CB1R from a molecular perspective can offer a glimpse of preventing potential toxicological or recreational effects since it is an attractive lead for drug development with fewer side effects than trans-THC.

Evidence for Disruption of Mg2+ Pair as a Resistance Mechanism Against HIV-1 Integrase Strand Transfer Inhibitors.

HIV-1 integrase is the enzyme responsible for integrating the viral DNA into the host genome and is one of the main targets for antiretroviral therapy; however, there are documented cases of resistance against all the currently used integrase strand transfer inhibitors (INSTIs). While some resistance-related mutations occur near the inhibitor's binding site, the mutation N155H occurs on the opposite side of the drug-interacting Mg2+ ions, thus, not interacting directly with the drug molecules and currently lacking an explanation for its resistance mechanism. Moreover, mutation N155H and the resistance-related mutation Q148H are mutually exclusive for unknown reasons. In the present study, we use molecular dynamics simulations to understand the impact of the N155H mutation in the HIV-1 integrase structure and dynamics, when alone or in combination with Q148H. Our findings suggest that the Mg2+ ions of the active site adopt different orientations in each of the mutants, causing the catalytic triad residues involved in the ion coordination to adapt their side-chain configurations, completely changing the INSTIs binding site. The change in the ion coordination also seems to affect the flexibility of the terminal viral DNA nucleotide near the active site, potentially impairing the induced-fit mechanism of the drugs. The explanations obtained from our simulations corroborate previous hypotheses drawn from crystallographic studies. The proposed resistance mechanism can also explain the resistance caused by other mutations that take place in the same region of the integrase and help uncover the structural details of other HIV-1 resistance mechanisms.