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The aim of our laboratory-based study was to investigate the extent of delayed-onset cell death after cryopreservation in endothelial and epithelial cell lines of ovarian origin. We found differences in percentages of vital cells directly after warming and after cultivation for 48 to 72 h. A granulosa cell line of endothelial origin (KGN) and an epithelial cell line (OvCar-3) were used. In both DMSO-containing and DMSO-free protocols, significant differences in vitality rates between the different cell lines when using open and closed vitrification could be shown (DMSO-containing: KGN open vs. OvCar open, p = 0.001; KGN closed vs. OvCar closed, p = 0.001; DMSO-free: KGN open vs. OvCar open, p = 0.001; KGN closed vs. OvCar closed, p = 0.031). Furthermore, there was a marked difference in the percentage of vital cells immediately after warming and after cultivation for 48 to 72 h; whereas the KGN cell line showed a loss of cell viability of 41% using a DMSO-containing protocol, the OvCar-3 cell loss was only 11% after cultivation. Using a DMSO-free protocol, the percentages of late-onset cell death were 77% and 48% for KGN and OvCar-3 cells, respectively. Our data support the hypothesis that cryopreservation-induced damage is cell type and cryoprotective agent dependent.

OBJECTIVES:. We sought to validate prognostic scores in coronavirus disease 2019 including National Early Warning Score, Modified Early Warning Score, and age-based modifications, and define their performance characteristics. DESIGN:. We analyzed prospectively collected data from the Adaptive COVID-19 Treatment Trial. National Early Warning Score was collected daily during the trial, Modified Early Warning Score was calculated, and age applied to both scores. We assessed prognostic value for the end points of recovery, mechanical ventilation, and death for score at enrollment, average, and slope of score over the first 48 hours. SETTING:. A multisite international inpatient trial. PATIENTS:. A total of 1,062 adult nonpregnant inpatients with severe coronavirus disease 2019 pneumonia. INTERVENTIONS:. Adaptive COVID-19 Treatment Trial 1 randomized participants to receive remdesivir or placebo. The prognostic value of predictive scores was evaluated in both groups separately to assess for differential performance in the setting of remdesivir treatment. MEASUREMENTS AND MAIN RESULTS:. For mortality, baseline National Early Warning Score and Modified Early Warning Score were weakly to moderately prognostic (c-index, 0.60–0.68), and improved with addition of age (c-index, 0.66–0.74). For recovery, baseline National Early Warning Score and Modified Early Warning Score demonstrated somewhat better prognostic ability (c-index, 0.65–0.69); however, National Early Warning Score+age and Modified Early Warning Score+age further improved performance (c-index, 0.68–0.71). For deterioration, baseline National Early Warning Score and Modified Early Warning Score were weakly to moderately prognostic (c-index, 0.59–0.69) and improved with addition of age (c-index, 0.63–0.70). All prognostic performance improvements due to addition of age were significant (p < 0.05). CONCLUSIONS:. In the Adaptive COVID-19 Treatment Trial 1 cohort, National Early Warning Score and Modified Early Warning Score demonstrated moderate prognostic performance in patients with severe coronavirus disease 2019, with improvement in predictive ability for National Early Warning Score+age and Modified Early Warning Score+age. Area under receiver operating curve for National Early Warning Score and Modified Early Warning Score improved in patients receiving remdesivir versus placebo early in the pandemic for recovery and mortality. Although these scores are simple and readily obtainable in myriad settings, in our data set, they were insufficiently predictive to completely replace clinical judgment in coronavirus disease 2019 and may serve best as an adjunct to triage, disposition, and resourcing decisions.

Twenty-five percent of U.S. schoolchildren attend a rural school. Yet, rural school issues are typically subsumed by debates focused on urban problems and the misguided notion of ample resources available for their remediation. These assumptions belie the reality of the spatial mismatch that exists for rural schools, especially around mental health supports. Adverse childhood experiences and trauma disproportionately affect rural schoolchildren, putting them at greater risk of academic underachievement and other negative throughout the lifespan. Trauma-informed approaches in rural schools may mitigate the effects of childhood adversity and help close achievement gaps for rural students. Rural schools and students have needs and challenges distinct from those of urban and suburban schools, but only 2% of peer-reviewed publications address trauma-informed approaches or social-emotional learning in rural schools. More research is needed to help our 13 million rural schoolchildren develop the resilience necessary to overcome adversity and achieve healthy outcomes.

With many advancements, technologies are now capable of recording non-human animals’ location, heart rate, and movement, often using a device that is physically attached to the monitored animals. However, to our knowledge, there is currently no technology that is able to do this unobtrusively and non-invasively. Here, we review the history of technology for use with animals, recent technological advancements, current limitations, and a brief introduction to our proposed novel software. Canadian tech mogul EAIGLE Inc. has developed an artificial intelligence (AI) software solution capable of determining where people and assets are within public places or attractions for operational intelligence, security, and health and safety applications. The solution also monitors individual temperatures to reduce the potential spread of COVID-19. This technology has been adapted for use at the Toronto Zoo, initiated with a focus on Sumatran orangutans (Pongo abelii) given the close physical similarity between orangutans and humans as great ape species. This technology will be capable of mass data collection, individual identification, pose estimation, behaviour monitoring and tracking orangutans’ locations, in real time on a 24/7 basis, benefitting both zookeepers and researchers looking to review this information.

Our objective was to identify the upper ambient temperature threshold that triggers an increase in cortisol in response to increased thermoregulatory demands in polar bears. The results reported here include endocrine data collected over two years from 25 polar bears housed in 11 accredited zoological institutions across North America. The effects of ambient temperature, sex, age group (juvenile, adult, elderly), breeding season and humidity on fecal cortisol metabolite (FCM) concentrations (N = 8439 samples) were evaluated using linear mixed models. Ambient temperatures were placed into five different categories: 20 °C. Ambient temperature and humidity had a significant (p < 0.05) effect on FCM concentrations with significant (p < 0.05) interactions of sex, age and breeding season. Once biotic factors were accounted for, there was a significant (p < 0.05) increase in FCM concentrations associated with ambient temperatures above 20 °C in adult polar bears. The implications of these findings for the management of both zoo and wild polar bears are discussed.

The Canadian beaver (Castor canadensis) is the largest indigenous rodent in North America. We report a draft annotated assembly of the beaver genome, the first for a large rodent and the first mammalian genome assembled directly from uncorrected and moderate coverage (< 30 ×) long reads generated by single-molecule sequencing. The genome size is 2.7 Gb estimated by k-mer analysis. We assembled the beaver genome using the new Canu assembler optimized for noisy reads. The resulting assembly was refined using Pilon supported by short reads (80 ×) and checked for accuracy by congruency against an independent short read assembly. We scaffolded the assembly using the exon–gene models derived from 9805 full-length open reading frames (FL-ORFs) constructed from the beaver leukocyte and muscle transcriptomes. The final assembly comprised 22,515 contigs with an N50 of 278,680 bp and an N50-scaffold of 317,558 bp. Maximum contig and scaffold lengths were 3.3 and 4.2 Mb, respectively, with a combined scaffold length representing 92% of the estimated genome size. The completeness and accuracy of the scaffold assembly was demonstrated by the precise exon placement for 91.1% of the 9805 assembled FL-ORFs and 83.1% of the BUSCO (Benchmarking Universal Single-Copy Orthologs) gene set used to assess the quality of genome assemblies. Well-represented were genes involved in dentition and enamel deposition, defining characteristics of rodents with which the beaver is well-endowed. The study provides insights for genome assembly and an important genomics resource for Castoridae and rodent evolutionary biology.

The grizzly bear (Ursus arctos ssp. horribilis) represents the largest population of brown bears in North America. Its genome was sequenced using a microfluidic partitioning library construction technique, and these data were supplemented with sequencing from a nanopore-based long read platform. The final assembly was 2.33 Gb with a scaffold N50 of 36.7 Mb, and the genome is of comparable size to that of its close relative the polar bear (2.30 Gb). An analysis using 4104 highly conserved mammalian genes indicated that 96.1% were found to be complete within the assembly. An automated annotation of the genome identified 19,848 protein coding genes. Our study shows that the combination of the two sequencing modalities that we used is sufficient for the construction of highly contiguous reference quality mammalian genomes. The assembled genome sequence and the supporting raw sequence reads are available from the NCBI (National Center for Biotechnology Information) under the bioproject identifier PRJNA493656, and the assembly described in this paper is version QXTK01000000.

The article presents the current conditions of abrasive water jet cutting process and factors relative to the quality of cutting surface. The main goal of research was to evaluate the assessment of the cutting depth, corrugated bottom cutting edge and roughness of the specimens depending on selected factors such as cutting speed and abrasive amount in the abrasive water jet process. Specimen were cut in four phases as a square. Main results were that the distance between water jet entering and water jet leaving is decreased with the increasing abrasive amount and by following lower cutting rates. The increasing of a cutting rate negatively effects the quality of the cut surface and the size of the distance between water jet entering and water jet leaving, because the increasing of a cutting rate increases also values of the mentioned parameters. As to the distance between water jet entering and water leaving, the abrasive amount of 200-250 g.min-1 at the rate of 50 mm.min-1 is considered to be optimal, but outside this range the influence of the abrasive amount impacts negatively, primarily on water jet entering and water jet leaving that has a direct influence on the corrugated bottom cutting edge.

To identify the clinical and epidemiological profile of cryptococcosis diagnosed at the University Hospital of the Federal University of Mato Grosso do Sul, Brazil, medical records of 123 patients admitted from January 1995 to December 2005 were analyzed. One hundred and four cases (84.5%) had HIV infection, six (4.9%) had other predisposing conditions and 13 (10.6%) were immunocompetent. Male patients predominated (68.3%) and their age ranged from 19 to 69 years (mean: 35.9). Most patients (73.2%) were born and lived lifelong in the state of Mato Grosso do Sul. Involvement of the central nervous system occurred in 103 patients (83.7%) and headache and vomiting were the most frequent symptoms. In 77 cases it was possible to identify the Cryptococcus species: 69 (89.6%) C. neoformans and eight (10.4%) C. gattii. Amphotericin B was the drug of choice for treatment (106/123), followed by fluconazole in 60% of cases. The overall lethality rate was 49.6%, being 51% among the HIV infected patients and 41.2% among the non-HIV infected (p > 0.05). Although cryptococcosis exhibited in our region a similar behavior to that described in the literature, the detection of an important rate of immunocompetent individuals and five C. gattii cryptococcosis in HIV-infected patients is noteworthy.O perfil clínico-epidemiológico de 123 casos de criptococose diagnosticados no Hospital Universitário da Universidade Federal de Mato Grosso do Sul, no período de janeiro de 1995 até dezembro de 2005, foi estudado retrospectivamente. Cento e quatro (84,9%) casos tinham associação com HIV, seis (4,9%) tinham outra condição predisponente e 13 (10,6%) eram imunocompetentes. Houve predomínio do sexo masculino (68,3%) e a idade variou de 19 a 69 anos (média de 35,9 anos). A maioria (73,2%) era natural e procedente de Mato Grosso do Sul. O envolvimento do sistema nervoso central ocorreu em 103 (83,7%) pacientes e os sintomas mais freqüentes foram cefaléia e vômitos. Em 77 casos foi possível identificar a espécie do agente, sendo 69 (89,6%) C. neoformans e oito (10,4%) C. gattii. O antifúngico mais utilizado foi anfotericina B (106/123) seguido de fluconazol em aproximadamente 60% dos casos. A taxa de letalidade foi de 49,6%, sendo 51% entre os pacientes infectados pelo HIV e 41,2% entre os não infectados pelo HIV (p > 0,005). Apesar da criptococose observada em nossa região apresentar comportamento semelhante ao descrito na literatura, chama a atenção a importante taxa da micose em imunocompetentes e cinco casos de infecção por C. gattii em pacientes HIV-positivos.