In vitro growth of human keratinocytes and oral cancer cells into microtissues: an aerosol-based microencapsulation technique

YesCells encapsulation is a micro-technology widely applied in cell and tissue research, tissue transplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue model for the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell line (ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 μL/min and air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginate microcapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 μm. Both cell lines were successfully grown into microtissues in the microcapsules of alginate within 16 days of culture. The microtissues were characterized by using a live/dead cell viability assay, field emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-plating experiments. The microtissues of both cell types were viable after being extracted from the alginate membrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrated differences in both nucleus size and morphology. The microtissues with re-associated cells in spheroids are potentially useful as a cell model for pharmacological studies.Malaysia Ministry of Education (Fundamental Research Grant Scheme, FRGS Vot. 1482 and IGSP Vot. 679)

In vitro growth of human keratinocytes and oral cancer cells into microtissues: an aerosol-based microencapsulation technique

YesCells encapsulation is a micro-technology widely applied in cell and tissue research, tissue transplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue model for the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell line (ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 μL/min and air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginate microcapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 μm. Both cell lines were successfully grown into microtissues in the microcapsules of alginate within 16 days of culture. The microtissues were characterized by using a live/dead cell viability assay, field emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-plating experiments. The microtissues of both cell types were viable after being extracted from the alginate membrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrated differences in both nucleus size and morphology. The microtissues with re-associated cells in spheroids are potentially useful as a cell model for pharmacological studies.Malaysia Ministry of Education (Fundamental Research Grant Scheme, FRGS Vot. 1482 and IGSP Vot. 679)

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

Prompt and non-prompt J/psi elliptic flow in Pb plus Pb collisions at root S-NN=5.02 TeV with the ATLAS detector

The elliptic flow of prompt and non-prompt J/ \u3c8 was measured in the dimuon decay channel in Pb+Pb collisions at sNN=5.02 TeV with an integrated luminosity of 0.42nb-1 with the ATLAS detector at the LHC. The prompt and non-prompt signals are separated using a two-dimensional simultaneous fit of the invariant mass and pseudo-proper decay time of the dimuon system from the J/ \u3c8 decay. The measurement is performed in the kinematic range of dimuon transverse momentum and rapidity 9 < pT< 30 GeV , | y| < 2 , and 0\u201360% collision centrality. The elliptic flow coefficient, v2, is evaluated relative to the event plane and the results are presented as a function of transverse momentum, rapidity and centrality. It is found that prompt and non-prompt J/ \u3c8 mesons have non-zero elliptic flow. Prompt J/ \u3c8v2 decreases as a function of pT, while for non-prompt J/ \u3c8 it is, with limited statistical significance, consistent with a flat behaviour over the studied kinematic region. There is no observed dependence on rapidity or centrality

Search for squarks and gluinos in final states with hadronically decaying tau-leptons, jets, and missing transverse momentum using pp collisions at root s = 13 TeV with the ATLAS detector

A search for supersymmetry in events with large missing transverse momentum, jets, and at least one hadronically decaying τ-lepton is presented. Two exclusive final states with either exactly one or at least two τ-leptons are considered. The analysis is based on proton-proton collisions at √s=13  TeV corresponding to an integrated luminosity of 36.1  fb⁻¹ delivered by the Large Hadron Collider and recorded by the ATLAS detector in 2015 and 2016. No significant excess is observed over the Standard Model expectation. At 95% confidence level, model-independent upper limits on the cross section are set and exclusion limits are provided for two signal scenarios: a simplified model of gluino pair production with τ-rich cascade decays, and a model with gauge-mediated supersymmetry breaking (GMSB). In the simplified model, gluino masses up to 2000 GeV are excluded for low values of the mass of the lightest supersymmetric particle (LSP), while LSP masses up to 1000 GeV are excluded for gluino masses around 1400 GeV. In the GMSB model, values of the supersymmetry-breaking scale are excluded below 110 TeV for all values of tanβ in the range 2 ≤ tanβ ≤ 60, and below 120 TeV for tanβ > 30.M. Aaboud … D. Duvnjak … P. Jackson … J.L. Oliver … A. Petridis … A. Qureshi … A.S. Sharma … M.J. White … et al. [The ATLAS Collaboration

5-Fluorouracil sensitizes colorectal tumor cells towards double stranded DNA breaks by interfering with homologous recombination repair

10.18632/oncotarget.3728Oncotarget61412574-1258

PARP1 inhibition radiosensitizes HNSCC cells deficient in homologous recombination by disabling the DNA replication fork elongation response.

There is a need to develop new, more efficient therapies for head and neck cancer (HNSCC) patients. It is currently unclear whether defects in DNA repair genes play a role in HNSCCs' resistance to therapy. PARP1 inhibitors (PARPi) were found to be "synthetic lethal" in cancers deficient in BRCA1/2 with impaired homologous recombination. Since tumors rarely have these particular mutations, there is considerable interest in finding alternative determinants of PARPi sensitivity. Effectiveness of combined irradiation and PARPi olaparib was evaluated in ten HNSCC cell lines, subdivided into HR-proficient and HR-deficient cell lines using a GFP-based reporter assay. Both groups were equally sensitive to PARPi alone. Combined treatment revealed stronger synergistic interactions in the HR-deficient group. Because HR is mainly active in S-Phase, replication processes were analyzed. A stronger impact of treatment on replication processes (p = 0.04) and an increased number of radial chromosomes (p = 0.003) were observed in the HR-deficient group. We could show that radiosensitization by inhibition of PARP1 strongly correlates with HR competence in a replication-dependent manner. Our observations indicate that PARP1 inhibitors are promising candidates for enhancing the therapeutic ratio achieved by radiotherapy via disabling DNA replication processes in HR-deficient HNSCCs