As COVID-19 was declared a health emergency in March 2020, there was immense demand for information about the novel pathogen. This paper examines the clinician-reported impact of Project ECHO COVID-19 Clinical Rounds on clinician learning. Primary sources of study data were Continuing Medical Education (CME) Surveys for each session from the dates of March 24, 2020 to July 30, 2020 and impact surveys conducted in November 2020, which sought to understand participants' overall assessment of sessions. Quantitative analyses included descriptive statistics and Mann-Whitney testing. Qualitative data were analyzed through inductive thematic analysis. Clinicians rated their knowledge after each session as significantly higher than before that session. 75.8% of clinicians reported they would 'definitely' or 'probably' use content gleaned from each attended session and clinicians reported specific clinical and operational changes made as a direct result of sessions. 94.6% of respondents reported that COVID-19 Clinical Rounds helped them provide better care to patients. 89% of respondents indicated they 'strongly agree' that they would join ECHO calls again.COVID-19 Clinical Rounds offers a promising model for the establishment of dynamic peer-to-peer tele-mentoring communities for low or no-notice response where scientifically tested or clinically verified practice evidence is limited.

Avian myelocytomatosis virus (MC29V) is a retrovirus that transforms both fibroblasts and macrophages in culture and induces myelocytomatosis, carcinomas, and sarcomas in birds. Previous work identified a sequence of about 1,500 nucleotides (here denoted onc(MCV)) that apparently derived from a normal cellular sequence and that may encode the oncogenic capacity of MC29V. In an effort to further implicate onc(MCV) in tumorigenesis, we used molecular hybridization to examine the distribution of nucleotide sequences related to onc(MCV) among the genomes of various avian retroviruses. In addition, we characterized further the genetic composition of the remainder of the MC29V genome. Our work exploited the availability of radioactive DNAs (cDNA's) complementary to onc(MCV) (cDNA(MCV)) or to specific portions of the genome of avian sarcoma virus (ASV). We showed that genomic RNAs of avian erythroblastosis virus (AEV) and avian myeloblastosis virus (AMV) could not hybridize appreciably with cDNA(MCV). By contrast, cDNA(MCV) hybridized extensively (about 75%) and with essentially complete fidelity to the genome of Mill Hill 2 virus (MH2V), whose pathogenicity is very similar to that of MC29V, but different from that of AEV or AMV. Hybridization with the ASV cDNA's demonstrated that the MC29V genome includes about half of the ASV envelope protein gene and that the remainder of the MC29V genome is closely related to nucleotide sequences that are shared among the genomes of many avian leukosis and sarcoma viruses. We conclude that onc(MCV) probably specifies the unique set of pathogenicities displayed by MC29V and MH2V, whereas the oncogenic potentials of AEV and AMV are presumably encoded by a distinct nucleotide sequence unrelated to onc(MCV). The genomes of ASV, MC29V, and other avian oncoviruses thus share a set of common sequences, but apparently owe their various oncogenic potentials to unrelated transforming genes.

Avian erythroblastosis virus (AEV) causes erythroblastosis and sarcomas in birds and transforms both erythroblasts and fibroblasts to neoplastic phenotypes in culture. The viral genetic locus required for oncogenesis by AEV is at present poorly defined; moreover, we know very little of the mechanism of tumorigenesis by the virus. To facilitate further analysis of these problems, we used molecular cloning to isolate the genome of AEV as recombinant DNA in a procaryotic vector. The identity of the isolated DNA was verified by mapping with restriction endonucleases and by tests for biological activity. The circular form of unintegrated AEV DNA was purified from synchronously infected quail cells and cloned into the EcoRI site of lambda gtWES x B. A restriction endonuclease cleavage map was established. By hybridization with complementary DNA probes representing specific parts of avian retrovirus genomes, the restriction map of the cloned AEV DNAs was correlated with a genetic map. These data show that nucleotide sequences unique to AEV comprise at least 50% of the genome and are located approximately in the middle of the AEV genome. Our data confirm and extend previous descriptions of the AEV genome obtained by other procedures. We studied in detail two recombinant clones containing AEV DNA: the topography of the viral DNA in the two clones was virtually identical, except that one clone apparently contained two copies of the terminal redundancy that occurs in linear viral DNA isolated from infected cells; the other clone probably contained only one copy of the redundant sequence. To recover infectious virus from the cloned DNA, we developed a procedure for transfection that compensated for the defectiveness of AEV in replication. We accomplished this by ligating cloned AEV DNA to the cloned DNA of a retrovirus (Rous-associated virus type 1) whose genome could complement the deficiencies of AEV. Ligation of the two viral DNAs was facilitated by using a neutral fragment of DNA as linker between otherwise noncompatible termini. Cloned AEV DNA gave rise to infectious AEV capable of transforming fibroblasts and bone marrow cells in culture and of inducing both sarcomas and erythroleukemia in chickens. We conclude that the cloned DNAs represent the authentic genome of AEV undisturbed by the cloning procedure. Molecular cloning offers a powerful approach to the identification and characterization of retrovirus genomes.

The retrovirus strain MC29 induces a variety of tumors in chickens, including myelocytomatosis and carcinomas of the kidney and liver. In addition, the virus can transform cultures of embryonic avian macrophages and fibroblasts. We have characterized the genome of MC29 virus and have identified nucleotide sequences that may encode the oncogenic potential ofthe virus. MC29 virus can replicate only with the assistance of a related helper virus. The defect in replication is apparently a consequence of a deletion in one or more viral genes: the haploid genome of the MC29 virus has a molecular weight of ca. 1.7 X 10(6), whereas the genome of the helper virus MCAV has a molecular weight of ca. 3.1 X 10(6). Although MC29 virus transforms fibroblasts in culture, its genome has no detectable homology with the gene src that is responsible for transformation of fibroblasts by avian sarcoma viruses. We prepared radioactive single-stranded DNA complementary to nucleotide sequences present in the genome of MC29 virus but not in the genome of MCAV (cDNA(MC29)). If they are contiguous, these sequences (ca. 1,500 nucleotides) are sufficiently complex to encode at least one protein. Homologous sequences were not detectable in several strains of avian sarcoma viruses or in an endogenous virus of chickens. Our findings confirm and extend recent reports from other laboratories and lead to the conclusion that MC29 virus may contain a previously unidentified gene(s) that is capable of transforming several distinct target cells. The evolutionary origins of this putative gene and its location on the viral genome can be explored with cDNA(MC29).

Cells producing Rous sarcoma virus contain virus-specific ribonucleic acid (RNA) which can be identified by hybridization to single-stranded deoxyribonucleic acid (DNA) synthesized with RNA-directed DNA polymerase. Hybridization was detected by either fractionation on hydroxyapatite or hydrolysis with single strand-specific nucleases. Similar results were obtained with both procedures. The hybrids formed between enzymatically synthesized DNA and viral RNA have a high order of thermal stability, with only minor evidence of mismatched nucleotide sequences. Virus-specific RNA is present in both nuclei and cytoplasm of infected cells. This RNA is remarkably heterogeneous in size, including molecules which are probably restricted to the nucleus and which sediment in their native state more rapidly than the viral genome. The nature of the RNA found in cytoplasmic fractions varies from preparation to preparation, but heterogeneous RNA (ca. 4-50 S ), smaller than the viral genome, is always present in substantial amounts.

The cytoplasm and nuclei of cells infected with Rous sarcoma virus contain RNA which can be annealed to single-stranded DNA synthesized by the RNA-dependent polymerase of the virus. Formation of DNA:RNA hybrids was detected by equilibrium centrifugation in Cs2SO4, and by step-wise elution from hydroxy-apatite. RNA which contains nucleotide sequences identical to those of the viral genome constitutes approximately 0.5% of the total RNA extracted from infected cells.

The products of the deoxyribonucleic acid (DNA) polymerase associated with Rous sarcoma virus and avian myeloblastosis virus were characterized by correlative analyses with equilibrium centrifugation and stepwise elution from hydroxyapatite. The initial enzymatic product consists of nascent DNA chains which are hydrogen-bonded to 70 S viral ribonucleic acid (RNA), whereas the final enzymatic product is double-stranded DNA. Appreciable amounts of free single-stranded DNA were not detected at any point during the course of the enzymatic reaction, but the data in this regard are not decisive. The time course of synthesis of DNA:RNA hybrids and double-stranded DNA has been analyzed. It is concluded that the synthesis of double-stranded DNA is a sequel to and is probably dependent upon the synthesis of DNA:RNA hybrid.

The deoxyribonucleic acid (DNA) polymerase of Rous sarcoma virus synthesizes both single- and double-stranded DNA, utilizing the ribonucleic acid (RNA) of the viral genome as the initial template. Results of pulse-chase experiments indicate that the single-stranded DNA serves as unconserved template and precursor for the synthesis of double-stranded DNA. The latter reaction is apparently initiated in association with the viral RNA and may involve a partially double-stranded intermediate form.

Purified preparations of Rous sarcoma virus (RSV) contain ribonuclease which is either a constituent of the virion surface or an adsorbed contaminant. Treatment of the virus with nonionic detergent to activate ribonucleic acid (RNA)-dependent deoxyribonucleic acid (DNA) polymerase renders the viral genome susceptible to hydrolysis by the external ribonuclease. The extent of this susceptibility can be substantially reduced by the use of limited amounts of detergent. At a concentration of detergent which provides a maximum initial rate of DNA synthesis, the degradation of endogenous viral RNA results in a reduced yield of high molecular weight DNA: RNA hybrid from the polymerase reaction. Attempts to detect virion-associated deoxyribonuclease, by using a variety of double helical DNA species as substrates, have been unsuccessful, but small amounts of nuclease activity directed against single-stranded DNA may be present in purified virus.

The deoxyribonucleic acid (DNA) polymerase(s) of Rous sarcoma virus synthesizes two principal products-single-stranded DNA in the form of a DNA:ribonucleic acid (RNA) hybrid and double-stranded DNA. All of the single-stranded product and 50% of the double-stranded product can be hybridized to 70S viral RNA. These results, in combination with data obtained by analysis of the kinetics of double-stranded DNA synthesis, indicate that the synthesis of double-stranded DNA is a sequel to the synthesis of single-stranded DNA and is dependent upon the latter for the provision of initial template.

Purified preparations of both the Bryan and Schmidt-Ruppin strains of Rous sarcoma virus contain two populations of low molecular weight RNA—a previously described 4 S RNA, which is probably transfer RNA derived from the host cell, and a newly identified 7 S RNA. The latter has been purified by chromatography on columns of methylated albumin kieselguhr, and has been found to be a single-stranded molecule with a molecular weight of approximately 80,000. It constitutes 3–5% of the total viral RNA, or 5–10 molecules per virion. The nucleotide composition of 7 S RNA is different for each strain of virus. The 7 S RNA is not detectably methylated, and thus is probably not a special form of transfer RNA. It has not been possible to identify a corresponding form of RNA in either normal or infected cells. Annealing experiments indicate that neither the 4 S nor the 7 S viral RNA is complementary to the RSV genome. The origin and significance of RSV 7 S RNA is presently indeterminate.

Deoxyribonucleic acid (DNA) polymerase activity can be elicited in purified preparations of avian myeloblastosis virus and Rous sarcoma virus (Schmidt-Ruppin strain) by treatment with nonionic detergent. The enzyme(s) and its synthetic products appear to be virion-associated. Enzymatic activity can be inhibited by pretreatment with either ribonuclease (8- to 10-fold inhibition) or actinomycin D (twofold inhibition). By contrast, rifampin has little, if any effect. The enzyme(s) synthesizes two primary products, a ribonucleic acid (RNA):DNA hybrid and DNA which is free of RNA. The results of both zonal and equilibrium centrifugation indicate that nascent chains of DNA are associated with the 70 S viral RNA. It is concluded that at least two enzymatic activities are under study: transcription of DNA from viral RNA, and subsequent, additional synthesis of DNA, utilizing product of the initial reaction as template.

The RNA-dependent and DNA-dependent DNA polymerase activities of RNA tumour viruses can be distinguished by their responses to actinomycin D; only the latter is inhibited.

A study was made of the pattern of congenital transmission of RIF, a naturally occurring avian leukosis virus, in a flock of chickens selected for a high incidence of neoplasms. It was found that about 1 out of every 6 hens had a persistent RIF-viremia, and all such hens infected their embryos regularly. One out of every 7 non-viremic hens was also a congenital transmitter of RIF, but the transmission was somewhat more erratic than in the case of the viremic birds. There was no indication that roosters, viremic or otherwise, could transmit RIF to their progeny, and it was concluded that the congenital transmission of RIF was strictly maternal.

Understanding household behavior and its macro consequences for society is pivotal for climate change adaptation. Yet, traditional policy decision-support models for nature–society systems oversimplify human behavior. Using original modeling ...Despite the growing calls to integrate realistic human behavior in sustainability science models, the representative rational agent prevails. This is especially problematic for climate change adaptation that relies on actions at various scales: from ...