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Axicabtagene ciloleucel or axi-cel [CD19 chimeric antigen receptor (CAR) T cell] has been recently approved for refractory/relapsed diffuse large B-cell lymphoma and primary mediastinal B-cell lymphoma. Proliferation of CAR T cells after infusion and their persistence have been reported as important factors. Laboratory tools are needed for the monitoring of patients. We developed a vector-based, simple, and accurate real-time quantitative PCR (qPCR) to measure axi-cel vector copy number in peripheral blood samples. Primers and probe targeting the 5'LTR region of the gammaretroviral vector (mouse stem cell virus) were designed for amplification. To generate standard curves, mouse stem cell virus plasmid was subcultured and quantified using droplet digital PCR. The method was applied to quantify vector copy number in blood samples from patients treated with axi-cel. The limit of quantification of the qPCR assay was established at 2.2 copies/μL in DNA eluate. The qPCR method was well correlated with flow cytometry findings; however, the assay appeared to be more sensitive than flow cytometry. The kinetics observed in blood samples from treated patients were in agreement with previously reported findings. In conclusion, we developed a sensitive and accurate qPCR assay for the quantification of transgenic CAR T cells, which can be a useful additional tool for the monitoring of patients treated with axi-cel.Post-transcriptional gene silencing mediated by microRNAs (miRNAs) modulates numerous developmental and stress response pathways. For the last two decades, HASTY (HST), the ortholog of human EXPORTIN 5, was considered to be a candidate protein that exports plant miRNAs from the nucleus to the cytoplasm. Here, we report that HST functions in the miRNA pathway independent of its cargo-exporting activity in Arabidopsis. We found that Arabidopsis mutants with impaired HST shuttling exhibit normal subcellular distribution of miRNAs. Interestingly, protein-protein interaction and microscopy assays showed that HST directly interacts with the microprocessor core component DCL1 through its N-terminal domain. this website Moreover, mass spectrometry analysis revealed that HST also interacts independently of its N-terminal domain with the mediator complex subunit MED37. Further experiments revealed that HST could act as a scaffold to facilitate the recruitment of DCL1 to genomic MIRNA loci by stabilizing the DCL1-MED37 complex, which in turn promotes the transcription and proper processing of primary miRNA transcripts (pri-miRNAs). Taken together, these results suggest that HST is likely associated with the formation of the miRNA biogenesis complex at MIRNA genes, promoting the transcription and processing of pri-miRNAs rather than the direct export of processed miRNAs from the nucleus.
Human immunodeficiency virus (HIV), hepatitis C virus (HCV) and hepatitis B virus (HBV) are substantial public health threats in the region of Central Asia and the Caucasus, where the prevalence of these infections is currently rising.
A systematic review of MEDLINE, Embase and PsycINFO was conducted with no publication date or language restrictions through October 2019. Additional data were also harvested from national surveillance reports, references found in discovered sources, and other "grey" literature. It included studies conducted on high-risk populations (people who inject drugs (PWID), female sex workers (FSW), men who have sex with men (MSM), prisoners, and migrants) in Central Asia Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, and Uzbekistan; and the Caucasus Armenia, Azerbaijan, Georgia, and Northern Caucasus region of the Russian Federation.
Wide ranges were noted for HIV prevalence PWID 0-30.1%, MSM 0-25.1%, prisoners 0-22.8%, FSW 0-10.0%, and migrants 0.06-1.5%, with the highest prevalence of these high-risk groups reported in Kazakhstan (for PWID), Georgia (for MSM and prisoners) and Uzbekistan (for migrants). HCV prevalence also had a wide range PWID 0.3-92.1%, MSM 0-18.9%, prisoners 23.8-49.7%, FSW 3.3-17.8%, and migrants 0.5-26.5%, with the highest prevalence reported in Georgia (92.1%), Kyrgyzstan (49.7%), and migrants from Tajikistan and Uzbekistan (26.5%). Similarly, HBV prevalence had a wide range PWID 2.8-79.7%, MSM 0-22.2%, prisoners 2.7-6.2%, FSW 18.4% (one study), and migrants 0.3-15.7%.
In Central Asia and the Caucasus, prevalence of HIV, HCV and HBV remains exceedingly high among selected populations, notably PWID and MSM.
In Central Asia and the Caucasus, prevalence of HIV, HCV and HBV remains exceedingly high among selected populations, notably PWID and MSM.
We evaluated the performance of the MDR/XDR-TB Colour Test (CT) as an in-house thin-layer agar-based indirect drug susceptibility test (DST) for Mycobacterium tuberculosis (MTB) in a non-expert setting in Estonia.
After 2 days of hands-on training for laboratory technicians, 6 panels of 150 MTB isolates were cultured onto CT plates prepared in-house in 2 laboratories. Triplicate readings of 900 CT plates resulted in 18 DST patterns for each initial isolate. Time intervals to the results and for media preparation were estimated, and intra- and interobserver agreement, test sensitivities and specificities were calculated. BACTEC MGIT 960 DST was used as a reference.
The median time to produce DST results for isoniazid, rifampicin and levofloxacin was 13 days. CT sensitivity was 94.7% for levofloxacin, 95.8% for isoniazid and 97.3% for rifampicin. Test specificities were >97% for all 3 drugs. Interobserver agreement was 100% in Lab A and in Lab B >97% for levofloxacin and 99% for isoniazid and rifampicin.
The implementation of the CT into a new laboratory was straightforward with only minimal guidance required. This study proves that the CT is highly reproducible and easily interpreted by previously inexperienced personnel.
The implementation of the CT into a new laboratory was straightforward with only minimal guidance required. This study proves that the CT is highly reproducible and easily interpreted by previously inexperienced personnel.
Canakinumab is an IL-1β antibody that neutralises the activity of IL-1β. This study examined the efficacy and safety of canakinumab in patients with moderate COVID-19-related pneumonia.
This study aimed to evaluate the reduction in duration of hospitalisation with adequate oxygen status. Forty-eight patients with moderate COVID-19-related pneumonia were asked to participate in the prospective case-control study 33 patients (cases) signed informed consent and received canakinumab (Cohort 1) and 15 patients (Controls) refused to receive the experimental drug and received institutional standard of care (Cohort 2).
Hospital discharge within 21 days was seen in 63% of patients in Cohort 1 vs. 0% in Cohort 2 (median 14 vs. 26 days, respectively; p < 0.001). There was significant clinical improvement in ventilation regimes following administration of canakinumab compared with Cohort 2 (Stuart-Maxwell test for paired data, p < 0.001). Patients treated with canakinumab experienced a significant increase in PaO
FiO
(p < 0.
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