Detection of SARS-CoV-2-Specific Humoral and Cellular Immunity in COVID-19 Convalescent IndividualsHighlights d SARS-CoV-2-specific antibodies are detected in COVID-19 convalescent subjects d Most COVID-19 convalescent individuals have detectable neutralizing antibodies d Cellular immune responses to SARS-CoV-2 are found in COVID-19 convalescent subjects d Neutralization antibody titers correlate with the numbers of virus-specific T cells.
Since 2007, many cases of fever, thrombocytopenia and leukopenia syndrome (FTLS) have emerged in Henan Province, China. Patient reports of tick bites suggested that infection could contribute to FTLS. Many tick-transmitted microbial pathogens were tested for by PCR/RT-PCR and/or indirect immunofluorescence assay (IFA). However, only 8% (24/285) of samples collected from 2007 to 2010 tested positive for human granulocytic anaplasmosis (HGA), suggesting that other pathogens could be involved. Here, we used an unbiased metagenomic approach to screen and survey for microbes possibly associated with FTLS. BLASTx analysis of deduced protein sequences revealed that a novel bunyavirus (36% identity to Tehran virus, accession: HQ412604) was present only in sera from FTLS patients. A phylogenetic analysis further showed that, although closely related to Uukuniemi virus of the Phlebovirus genus, this virus was distinct. The candidate virus was examined for association with FTLS among samples collected from Henan province during 2007–2010. RT-PCR, viral cultures, and a seroepidemiologic survey were undertaken. RT-PCR results showed that 223 of 285 (78.24%) acute-phase serum samples contained viral RNA. Of 95 patients for whom paired acute and convalescent sera were available, 73 had serologic evidence of infection, with 52 seroconversions and 21 exhibiting a 4-fold increase in antibody titer to the virus. The new virus was isolated from patient acute-phase serum samples and named Henan Fever Virus (HNF virus). Whole-genome sequencing confirmed that the virus was a novel bunyavirus with genetic similarity to known bunyaviruses, and was most closely related to the Uukuniemi virus (34%, 24%, and 29% of maximum identity, respectively, for segment L, M, S at maximum query coverage). After the release of the GenBank sequences of SFTSV, we found that they were nearly identical (>99% identity). These results show that the novel bunyavirus (HNF virus) is strongly correlated with FTLS.
We propose that these α/β core neurons serve as a "gate" to keep LTM from being formed, whereas any experience capable of "opening" the gate is given permit to be consolidated into LTM.
Unlike other solid organ transplants, intestinal transplantation (IT) remains a highly experimental procedure. Rejection, sepsis, and graft-versus-host disease have been the major barriers to successful IT in humans. These problems can be studied in the rat model, but this requires a reliable surgical technique that will produce high survival rates and excellent graft function. Herein, we review 400 consecutive IT in rats and describe important technical aspects of surgery. Technical modifications that have helped to reduce the morbidity after IT in rats include 1) minimizing mechanical and ischemic injuries to grafts during the donor procedure, 2) marking the portal vein and aortic conduit with sutures to ensure correct orientation of the graft, 3) using a macaroni noodle to stent the intestinal anastomosis, and 4) administering large volumes of crystalloid to maintain a normal blood pressure during the donor and recipient surgeries. The survival rate in 298 rats with accessory, heterotopic grafts was 90%. The survival rate in 102 rats with orthotopic (in continuity) graft (OIT) was 86%. Rats have survived more than 500 days after OIT, maintaining normal weights, intestinal function, and intestinal histology.
Efficient intracellular delivery of biologically active macromolecules has been a challenging but important process for manipulating live cells for research and therapeutic purposes. There have been limited transfection techniques that can deliver multiple types of active molecules simultaneously into single-cells as well as different types of molecules into physically connected individual neighboring cells separately with high precision and low cytotoxicity. Here, a high frequency ultrasound-based remote intracellular delivery technique capable of delivery of multiple DNA plasmids, messenger RNAs, and recombinant proteins is developed to allow high spatiotemporal visualization and analysis of gene and protein expressions as well as single-cell gene editing using clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9), a method called acoustic-transfection. Acoustic-transfection has advantages over typical sonoporation because acoustic-transfection utilizing ultra-high frequency ultrasound over 150 MHz can directly deliver gene and proteins into cytoplasm without microbubbles, which enables controlled and local intracellular delivery to acoustic-transfection technique. Acoustic-transfection was further demonstrated to deliver CRISPR-Cas9 systems to successfully modify and reprogram the genome of single live cells, providing the evidence of the acoustic-transfection technique for precise genome editing using CRISPR-Cas9.
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