The coronavirus disease 2019 (COVID-19) pandemic continues to spread globally, highlighting the urgent need for safe and effective vaccines that could be rapidly mobilized to immunize large ...populations. We report the preclinical development of a self-amplifying mRNA (SAM) vaccine encoding a prefusion stabilized severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein and demonstrate strong cellular and humoral immune responses at low doses in mice and rhesus macaques. The homologous prime-boost vaccination regimen of SAM at 3, 10 and 30 μg induced potent neutralizing antibody (nAb) titers in rhesus macaques following two SAM vaccinations at all dose levels, with the 10 μg dose generating geometric mean titers (GMT) 48-fold greater than the GMT of a panel of SARS-CoV-2 convalescent human sera. Spike-specific T cell responses were observed with all tested vaccine regimens. SAM vaccination provided protective efficacy against SARS-CoV-2 challenge as both a homologous prime-boost and as a single boost following ChAd prime, demonstrating reduction of viral replication in both the upper and lower airways. The SAM vaccine is currently being evaluated in clinical trials as both a homologous prime-boost regimen at low doses and as a boost following heterologous prime.
ONYX-015 is an adenovirus that lacks the E1B-55K gene product for p53 degradation. Thus, ONYX-015 was conceived as an oncolytic virus that would selectively replicate in p53-defective tumor cells. ...Here we show that loss of E1B-55K leads to the induction, but not the activation, of p53 in ONYX-015-infected primary cells. We use a novel adenovirus mutant, ONYX-053, to demonstrate that loss of E1B-55K-mediated late viral RNA export, rather than p53 degradation, restricts ONYX-015 replication in primary cells. In contrast, we show that tumor cells that support ONYX-015 replication provide the RNA export function of E1B-55K. These data reveal that tumor cells have altered mechanisms for RNA export and resolve the controversial role of p53 in governing ONYX-015 oncolytic selectivity.
We have engineered a human adenovirus, ONYX-411, that selectively replicates in human tumor cells, but not normal cells, depending upon the status of their retinoblastoma tumor suppressor protein ...(pRB) pathway. Early and late viral gene expression as well as DNA replication were significantly reduced in a functional pRB-pathway-dependent manner, resulting in a restricted replication profile similar to that of nonreplicating adenoviruses in normal cells both in vitro and in vivo. In contrast, the viral life cycle and tumor cell killing activity of ONYX-411 was comparable to that of wild-type adenovirus following infection of human tumor cells in vitro as well as after systemic administration in tumor-bearing animals.
Therapeutic vaccines that elicit cytotoxic T cell responses targeting tumor-specific neoantigens hold promise for providing long-term clinical benefit to patients with cancer. Here we evaluated ...safety and tolerability of a therapeutic vaccine encoding 20 shared neoantigens derived from selected common oncogenic driver mutations as primary endpoints in an ongoing phase 1/2 study in patients with advanced/metastatic solid tumors. Secondary endpoints included immunogenicity, overall response rate, progression-free survival and overall survival. Eligible patients were selected if their tumors expressed one of the human leukocyte antigen-matched tumor mutations included in the vaccine, with the majority of patients (18/19) harboring a mutation in KRAS. The vaccine regimen, consisting of a chimp adenovirus (ChAd68) and self-amplifying mRNA (samRNA) in combination with the immune checkpoint inhibitors ipilimumab and nivolumab, was shown to be well tolerated, with observed treatment-related adverse events consistent with acute inflammation expected with viral vector-based vaccines and immune checkpoint blockade, the majority grade 1/2. Two patients experienced grade 3/4 serious treatment-related adverse events that were also dose-limiting toxicities. The overall response rate was 0%, and median progression-free survival and overall survival were 1.9 months and 7.9 months, respectively. T cell responses were biased toward human leukocyte antigen-matched TP53 neoantigens encoded in the vaccine relative to KRAS neoantigens expressed by the patients' tumors, indicating a previously unknown hierarchy of neoantigen immunodominance that may impact the therapeutic efficacy of multiepitope shared neoantigen vaccines. These data led to the development of an optimized vaccine exclusively targeting KRAS-derived neoantigens that is being evaluated in a subset of patients in phase 2 of the clinical study. ClinicalTrials.gov registration: NCT03953235 .
Structured reporting (SR) replaced narrative (free text) reporting and utilizes templated headings and subheadings with findings typically based on the anatomy included in the examination. Its use ...has been widely advocated by radiology and non-radiology organizations as the new reporting standard. There are, however, shortcomings to SR, such as templated text not addressing a specific clinical indication. Contextual reporting (CR) fills this gap. CR is a type of SR which is tailored to a narrow clinical indication by including pertinent positive and negative findings for that specific clinical entity.
This study assesses provider preferences for CR as compared to SR in the pediatric practice environment using a survey methodology.
Surveys with examples of SR and CR reports were sent electronically to two groups. One group was focused on neurological diseases and included pediatric specialists in neurosurgery, neurology, ENT, ED, and ophthalmology (190 people), referred to as the pediatric neuroimaging group. The pediatric neuroimaging group survey contained examples of CR and SR reports of an orbital CT for orbital cellulitis and a head CT for stroke. The other group was focused on gastrointestinal diseases, and included pediatric specialists in gastroenterology, general surgery, and the ED (159 people), referred to as the pediatric gastrointestinal (GI) imaging group. The pediatric GI imaging group survey contained example reports of an abdominal CT for appendicitis and an MRI enterography for Crohn's disease. Surveys utilizing a 5-point Likert scale were analyzed via Fischer's exact test with a p-value deemed statistically significant at less than 0.05.
349 individuals were contacted to participate in the survey. There were 81 (23 %, 81/349) survey respondents; 41 (22 %, 41/190) from the neuro group, and 40 (25 %, 40/159) from the GI group. 56 % (45/81) of all respondents preferred CR reports over traditional SR reports, while 29 % (23/81) did not. Most respondents (59 %, 48/81) indicated that CR reports are easier to interpret than traditional SR reports. Respondents from the pediatric neuroimaging group favored CR reports to a lesser degree (44 %, 36/81) compared to respondents from the pediatric GI imaging group (68 %, 55/81).
We learned from this survey that it would be beneficial to be very intentional about selecting clinical indications where CR would be most valued rather than trying to develop CR for any specific clinical indication. The study results indicate it is reasonable to continue further efforts at exploring the utility of contextualized reports.
Lengthy MRI examinations in young children often requires sedation. When sedation is unavailable, critical imaging may be delayed. Abbreviating the imaging protocol to a few essential sequences may ...reduce the need for sedation and prevent delays in patient care. We retrospectively evaluated an abbreviated noncontrast MRI protocol to diagnose lower extremity osteomyelitis in the pediatric population. The IRB approved this study. The radiology information system was searched for lower extremity MRI examinations for osteomyelitis in patients <20 years old from August 2020 to August 2021. Three noncontrast sequences (long axis T1 without fat saturation (FS), long axis STIR, and axial T2 with FS) were independently reviewed by 2 pediatric radiologists. The accuracy of the reviewers was compared to the clinical radiology report based on the unabridged contrast-enhanced standard department protocol. The search yielded 80 exams, mean age was 7 years old, 59% (47/80) were male, and 41% (33/80) were female. The accuracies of reviewer A and reviewer B were 95% and 89%, respectively. The reviewer inter-observer agreement for the diagnosis of osteomyelitis was strong (k = 0.79). The accuracy of an abbreviated noncontrast MRI protocol to evaluate lower extremity osteomyelitis in children approaches that of the unabridged protocol and has the potential to decrease the need for sedation in young children.
In interspecific hybrids or allopolyploids, often one parental set of ribosomal RNA genes is transcribed and the other is silent, an epigenetic phenomenon known as nucleolar dominance. Silencing is ...enforced by cytosine methylation and histone deacetylation, but the initial discrimination mechanism is unknown. One hypothesis is that a species-specific transcription factor is inactivated, thereby silencing one set of rRNA genes. Another is that dominant rRNA genes have higher binding affinities for limiting transcription factors. A third suggests that selective methylation of underdominant rRNA genes blocks transcription factor binding. We tested these hypotheses using Brassica napus (canola), an allotetraploid derived from B. rapa and B. oleracea in which only B. rapa rRNA genes are transcribed. B. oleracea and B. rapa rRNA genes were active when transfected into protoplasts of the other species, which argues against the species-specific transcription factor model. B. oleracea and B. rapa rRNA genes also competed equally for the pol I transcription machinery in vitro and in vivo. Cytosine methylation had no effect on rRNA gene transcription in vitro, which suggests that transcription factor binding was unimpaired. These data are inconsistent with the prevailing models and point to discrimination mechanisms that are likely to act at a chromosomal level.
In this report, we describe a vector system that specifically delivers transgene products to tumors following intravenous (i.v.) administration. The Escherichia coli cytosine deaminase (CD) gene was ...placed in the E3B region of the tumor-selective, replication-competent adenovirus ONYX-411, under the control of endogenous viral late gene regulatory elements. Thus, CD expression was directly coupled to the tumor-selective replication of the viral vector. In vitro, CD was expressed efficiently in various human cancer cell lines tested but not in cultured normal human cells, including human hepatocytes. Following i.v. administration into nude mice carrying human tumor xenografts, robust CD activity was detected only in tumors but not in liver or other normal tissues. Levels of CD activity in the tumors increased progressively following i.v. virus administration, correlating closely with virus replication in vivo. Subsequent administration of 5-fluorocytosine (5-FC) demonstrated a trend to improve the antitumor efficacy of these viruses in a mouse xenograft model, presumably due to the intratumoral conversion of 5-FC to the chemotherapeutic drug 5-fluorouracil. We show that the combination of a highly selective oncolytic virus, ONYX-411, with the strategic use of the viral E3B region for transgene insertion provides a powerful platform that allows for tumor-specific, persistent and robust transgene expression after i.v. administration. This technology provides an opportunity to enhance greatly both safety and efficacy of cancer gene therapy.
One of the challenges to secure wireless sensor networks (WSNs) is to design secure pair-wise key agreement between any pair of resources-limited sensors while keeping strong connectivity. In this ...paper, we point out the fatal security weaknesses of Cheng-Agrawal's pair-wise key agreement scheme for WSNs, and then propose a new scheme. Compared to the existing schemes, the proposed scheme owns two outstanding merits- assurance of connectivity between any pair of nodes and security robustness against node capture attack.
Mineral compositions of aerosol particles were investigated at four sites （Aksu, Dunhuang, Zhenbeitai, and Tongliao） in desert regions of northern China from March to May in 2001 and 2002 during the ...intensive field campaign period of ACE-Asia （Aerosol Characterization Experiments-Asia）. The X-ray diffraction （XRD） results show the main minerals for Asian dust are illite, chlorite, kaolinite, quartz, feldspar, calcite, and dolomite. Gypsum, hornblende, and halite are also detected in several samples. Semi-quantitative mineralogical data of aerosol samples show that carbonate content decreases from western to eastern source areas; that is, soil dust collected at western source area sites of Dunhuang and Aksu are enriched with carbonate, while northeastern source area site of Tongliao is associated with low carbonate content. But the spatial distribution of feldspar exhibits a different pattern as compared to carbonate, increasing from the western to the eastern sources. The total clay content is significantly higher （73% in average） at the deposition site of Changwu than those at source areas. Air-mass back trajectory studies for the three dust storm events observed at Changwu, showed that soil dust transport pathways were as expected from carbonate content for the source identification, further demonstrating that carbonate was a useful tracer for eolian dust on regional scale in northern China.
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