Extracellular vesicles (EVs) can transport molecules that combat viruses, such as RNA against SARS-CoV-2. Bacterial coinfections can help establish certain viruses and worsen diseases. Thus, we designed a model to induce the secretion of polydisperse EVs shown with SARS-CoV-2 and bacterial coinfection using macrophages and E. coli fractions as in vitro inducers. We obtained short and large macrophage EVs. The E. coli fraction was designated as SDS-soluble bacterial membrane fraction and its associated proteins (SDS-SBMF). The proteins were identified using a mass spectrometer. SDS-SBMF contained mainly OmpF, OmpA, OmpC, OmpX, and lpp. The SDS-SBMF macrophages induced the secretion of polydisperse EVs at 30 min, reaching optimal secretion at 120 min, as observed via scanning electron microscopy and confocal microscopy. Macrophage EVs contained mainly HSP7C, actin, apolipoprotein, GAPDH, annexin A5, PKM, moesin, and cofilin. We observed an increase in EVs in the bloodstream of patients with SARS-CoV-2 and bacterial coinfection, in addition to the presence of SARS-CoV-2 genes (E, ORF) in EVs. This in vitro method for inducing EVs has the potential to be used to obtain larger samples for study and for the detection of diagnostic and prognostic biomarkers of different diseases.

Ovarian cancer (OC) is diagnosed at an advanced stage, contributing to its high mortality rate. The presence of concurrent bacterial infections in these patients is a common clinical observation, and the mechanisms by which this coinfection influences tumor progression, still not fully understood. This study investigates the role of extracellular vesicles polydispersity (EVPs) secreted by OC cells in response to bacterial components, aiming to elucidate a potential communication pathway between the OC and the bacterial microenvironment. We stimulated a human OC cell line in vitro with a fraction of E. coli. Our results show that this bacterial stimulation significantly increases the secretion of PEVs by the cancer cells. A subsequent proteomic analysis of these PEVs revealed an enrichment of proteins, including filamin A-B, alpha-enolase, heat shock cognate 71 kDa protein, etc. In addition, the EVPs displayed protease (on fibronectin and gelatin) and phosphatase activity against para-nitrophenyl phosphate, indicating their capacity to alter cellular signaling. This represents a novel mechanism through which bacterial coinfection may influence the biological behavior of OC, potentially contributing to their aggressiveness and the challenges associated with their treatment. Our work highlights the importance of studying the interplay between the tumor and its associated microbiota to better understand ovarian cancer progression and identify new therapeutic targets.

Background/Objectives: This study evaluated variation in nodal support for provisional HIV-1 subtype B compatibility across pol, gag, vif, tat and vpr in an institutional cohort from central Mexico. Methods: We performed a secondary descriptive bioinformatics reanalysis of 60 treatment-naïve adults. Paired FASTQ files were available for 58 samples, and 42 were retained across all five regions. Robust support required SH-aLRT ≥ 80 and UFBoot ≥ 95. Additional analyses varied reference-composition thresholds, reselected assignment nodes, harmonized the reference panel and subsampled alignment columns. Results: At the originally recorded nodes, 18/210 sample–region records (8.6%) were robust; 25/42 samples had no robust region, 16 had one and one had two. Node reselection at the 60% composition threshold retained all 18 records and added S42/pol and S29/tat, yielding 20/210 (9.5%). Neither analysis identified robust support in three or more regions. With a common reference panel, the ordering of contextual internal-branch support between pol and vif changed. Conclusions: Support depended on the evaluated node and analytical dataset. Fixed-node threshold stability did not establish stability under node reselection. Differences among regional analyses could not be attributed independently to genomic region. Single-fragment assignments remain provisional and do not establish whole-genome subtype classifications. V3 and ViReMa analyses were exploratory.

Histiocytic sarcoma (HS) is an aggressive hematological malignancy whose transformed cells exhibit morphological and immunophenotypic characteristics similar to macrophages, and arises de novo or as part of a clonal ‘evolution’ of other pre-existing hematological neoplasms. This study investigates the potential use of the J774A.1 cell line (a cell line derived from murine tumor cells, commonly used in macrophage research) as a research model to study the role of polydisperse extracellular vesicles (PEVs) secreted by the HS cells, considering that bacterial infections are common in patients with cancer, including HS. The influences of bacterial components on tumor progression are still not fully understood. We stimulated the J774A.1 cell line in vitro with a fraction of E. coli, and our results show that the bacterial stimulation increases the secretion of PEVs by these cells. Comparative results of J774A.1 cells with PEVs using confocal and scanning electron microscopy with micrographic reports of HS histological slides (from several cited mammal species, including humans) suggest a possible relationship of large PEVs with marks, footprints, or traces of possible large PEVs disrupted in the HS of these reports. A subsequent proteomic analysis of these PEVs revealed a diverse subcellular origin of their components, such as proteins including: Triosephosphate isomerase (TPI), Heat shock cognate 71 kDa, Apolipoprotein A-1, Rho GDP-dissociation inhibitor 1, GAPDH, Galectin, Moesin, globular Actin, and Annexin. These results highlight the importance of studying the interplay between HS, other hematological cancers, and bacterial infections to better understand the progression of this cancer, identify new therapeutic targets, and emphasize the importance of preventing bacterial infections in cancer patients. Furthermore, the results demonstrate the potential use of the stimulated J774A.1 cell line for research on HS-related PEVs.

Histiocytic sarcoma (HS) is an aggressive hematological malignancy whose trans-formed cells exhibits morphological and immunophenotypic characteristics similar to macrophages, and arise de novo, or as part of a clonal ‘‘evolution’’ of other pre-existing hematological neoplasms. This study investigates the potential use of the J774A.1 cell line (a line derived from murine tumor cells, commonly used in macrophage research) as a research model for the role of polydisperse extracellular vesicles (PEVs) secreted by the HS cells, considering that bacterial infections are common in patients with cancer, including HS. The influences of bacterial components on tumor progression are still not fully understood. We stimulated the J774A.1 cell line in vitro with a fraction of E. coli and our results show that the bacterial stimulation increases the secretion of PEVs by these cells. Comparative results of J774A.1 cells with PEVs using confocal and scanning electron microscopy with micrographic reports of HS histological slides (from several cited mammal species, including humans) suggest a possible relationship of large PEVs with marks, footprints or traces of possible large PEVs disrupted in the HS of these reports. A subsequent proteomic analysis of these PEVs revealed a diverse subcellular origin of its components such as proteins as including: Triosephosphate isomerase (TPI), Heat shock cognate 71 kDa, Apolipoprotein A-1, Rho GDP-dissociation inhibitor 1, GAPDH, Galectin, Moesin, globular Actin and Annexin. These results highlight the importance of studying the interplay between the HS, others hematological cancers, and bacterial infections to better understand progression of this cancer, identify new therapeutic targets and emphasize the importance of preventing bacterial infections in cancer patients. Furthermore, the results demonstrate the potential use of the stimulated J774A.1 cell line for research of the HS-related PEVs.

Ovarian cancer (OC) is usually diagnosed at an advanced stage, contributing to its high mortality rate. The presence of concurrent bacterial infections in these patients is a common clinical observation, and the mechanisms by which this coinfection influences tumor progression are still not fully understood. This study investigates the role of polydisperse extracellular vesicles (PEVs) secreted by OC cells in response to bacterial components, aiming to elucidate a potential communication pathway between OC and the bacterial microenvironment. We stimulated a human OC cell line in vitro with a fraction of E. coli. Our results show that this bacterial stimulation significantly increases the secretion of PEVs by cancer cells. A subsequent proteomic analysis of these PEVs revealed an enrichment of proteins, including filamin A, filamin B, alpha-enolase, and heat shock cognate 71 kDa protein. In addition, the PEVs displayed protease activity (on fibronectin and gelatin) and phosphatase activity against para-nitrophenyl phosphate, indicating their capacity to alter cellular signaling. This represents a novel mechanism through which bacterial coinfection may influence the biological behavior of OC if bacteria interact with tumor cells, potentially contributing to their aggressiveness and the challenges associated with their treatment. Our work highlights the importance of studying the interplay between the tumor and its associated microbiota to better understand ovarian cancer progression and identify new therapeutic targets.