Reading Through the Pause: How Superintendents Viewed Literacy for Middle Grade Learners During the Pandemic

This paper highlights the voices of two superintendents\u27 lived experiences guiding teachers, parents, and students in their districts during the pandemic shutdown. The emphasis of literacy education showcases the ways in which middle grades learners were able to continue discursive practices through online platforms to share and engage with texts. This reflective piece describes the process of perseverance in literacy education through the pandemic pause

Human chronic wounds treated with bioengineered skin: Histologic evidence of host-graft interactions

Bioengineered skin is being used to successfully treat a variety of wounds. Randomized controlled clinical trials have shown that a living bilayered skin construct (BSC), consisting of human neonatal keratinocytes and fibroblasts in a collagen matrix, was able to accelerate complete closure of both venous and diabetic ulcers. BSC was particularly effective in difficult-to-heal wounds of long duration. In patients treated with BSC, no obvious signs of gross clinical rejection were observed. Testing of these treated patients showed no BSC-specific immune response and no immune response to bovine collagen or alloantigens expressed on keratinocytes and fibroblasts. However, very little is known about the histologic changes that occur after BSC has been placed on human wounds. We report our preliminary histologic observations in this uncontrolled study of a cohort of 11 patients with 14 wounds treated with BSC in whom biopsy specimens of the grafted sites were obtained at least 2 weeks after application of the construct. The etiology of these ulcers varied from arterial or venous disease to an extensively and poorly healing burn wound. Histologically, thickening of the grafted bioengineered skin was seen in all samples where residual BSC could be identified. Mucin deposition was noted in the dermal layer of the wounds and BSC in 13 of the 14 specimens examined. Unexpectedly, and in spite of good clinical outcome, 4 of the 14 specimens exhibited a foreign body-like granulomatous response. There was no history of prior exposure to BSC in the 4 patients who had a granulomatous response. These early histologic observations suggest that stimulatory interactions develop between BSC and the wound. The consistently found deposition of mucin may point to a fetal pattern of wound repair associated with the neonatal cells in BSC. (J Am Acad Dermatol 2002;46:524-30.

The role of respondent-driven sampling in the behavioural surveillance of people who inject drugs

People who inject drugs (PWID) are greatly affected by blood borne viruses (BBV). Behavioural surveillance, which consists of ongoing cross-sectional surveys of at-risk populations, is a core component of BBV prevention. Respondent-driven sampling (RDS) is a relatively new sampling method, developed to sample hard-to-reach populations, such as PWID. This thesis aimed to describe the role of RDS in the behavioural surveillance of PWID. An RDS study of PWID was conducted in 2009 in Sydney, Australia. Five seeds were selected to start the recruitment, which led to a sample of 261 participants over 16 waves. To achieve the thesis aim, I examined the benefits and challenges of conducting RDS with minimal and existing resources, the ability to use RDS data to calculate unbiased population estimates, the characteristics of the RDS study’s sample as compared with those from separate service-based sampling studies, and whether RDS network data can add to the usefulness of a behavioural surveillance system. While there were benefits to conducting RDS with existing resources, there were also challenges related to crowd control and privacy issues. An examination of RDS requirements and assumptions found a lack of random recruitment and a low proportion of coupons returned, which suggested that population estimates were biased. In a comparison of RDS participants with participants from two separate service-based sampling studies, no marked differences in demographics or risk practices were found. As well, compared to pharmacy-based sampling, RDS was 80% less likely to capture participants with small injecting networks, a group found to be of interest to behavioural surveillance. Finally, using the network information gathered through RDS, it was found that closeness, Aboriginal origin status and hepatitis C testing were related to sharing injecting equipment within dyads, which were formed by linking participants’ information with that of their recruiter. The results indicate that in locations where there is good coverage by services for PWID, RDS should be best viewed as a complementary tool, where it can be used on an intermittent basis to evaluate existing sampling methods and to add to the understanding of how network characteristics shape risk practices

Wounding of Bioengineered Skin: Cellular and Molecular Aspects After Injury

Skin substitutes are increasingly being used in the treatment of difficult to heal wounds but their mechanisms of action are largely unknown. In this study, using histology, immunostaining, flow cytometry, enzyme-linked immunosorbent assay, and reverse transcription polymerase chain reaction, we determined the response to injury of a human bilayered skin substitute. Meshing or scalpel fenestration of the construct was found to stimulate keratinocyte migration and to decrease proliferation. By 24 h, flow cytometry of the keratinocyte component showed that meshing was associated with a 33% decrease in the number of cells in S phase (p <0.01). An approximately 2-fold decrease in staining for Ki67, a proliferation marker, was observed with meshing of human bilayered skin substitute. The process of reepithelialization was apparent by 12 h, however, the wounded human bilayered skin substitute was healed by day 3, and a stratum corneum and fully stratified epithelium were re-established by day 4. Reverse transcription polymerase chain reaction analysis and enzyme-linked immunosorbent assays showed that the expression of acute proinflammatory cytokines (interleukins 1α, 6, and 8, tumor necrosis factor α) peaked by 12–24 h postinjury. The levels of mRNA of certain growth factors (transforming growth factor β1, vascular endothelial growth factor, insulin-like growth factor 2) but not others (platelet-derived growth factors A and B, keratinocyte growth factor, fibroblast growth factors 1 and 7, transforming growth factor β3) increased by 12 h and peaked by 1–3 d after injury, returning to normal by day 6. Immunostaining for tumor necrosis factor α and transforming growth factor β1 paralleled these findings by reverse transcription polymerase chain reaction. We conclude that human bilayered skin substitute, as a prototypic bilayered skin substitute, is a truly dynamic living tissue, capable of responding to physical injury in a staged and specific pattern of cell migration, reepithelialization, and cytokine expression

2C-ChIP: measuring chromatin immunoprecipitation signal from defined genomic regions with deep sequencing

Abstract Background Understanding how transcription occurs requires the integration of genome-wide and locus-specific information gleaned from robust technologies. Chromatin immunoprecipitation (ChIP) is a staple in gene expression studies, and while genome-wide methods are available, high-throughput approaches to analyze defined regions are lacking. Results Here, we present carbon copy-ChIP (2C-ChIP), a versatile, inexpensive, and high-throughput technique to quantitatively measure the abundance of DNA sequences in ChIP samples. This method combines ChIP with ligation-mediated amplification (LMA) and deep sequencing to probe large genomic regions of interest. 2C-ChIP recapitulates results from benchmark ChIP approaches. We applied 2C-ChIP to the HOXA cluster to find that a region where H3K27me3 and SUZ12 linger encodes HOXA-AS2, a long non-coding RNA that enhances gene expression during cellular differentiation. Conclusions 2C-ChIP fills the need for a robust molecular biology tool designed to probe dedicated genomic regions in a high-throughput setting. The flexible nature of the 2C-ChIP approach allows rapid changes in experimental design at relatively low cost, making it a highly efficient method for chromatin analysis

Use of respondent-driven sampling to enhance understanding of injecting networks: a study of people who inject drugs in Sydney, Australia

BackgroundRespondent-driven sampling (RDS) is a method for recruiting hidden populations, such as people who inject drugs (PWID). In RDS, participants recruit their peers into the study; who recruited who into the study is tracked, and thus information is gathered on the population's social networks. The purpose of this study was to use information collected from an RDS study of PWID to determine the size and structure of injecting networks and whether network characteristics are associated with sharing injecting equipment.MethodsA study was launched in Sydney, Australia in 2009 with five seeds, who were asked to recruit three participants each into the survey. This process was repeated until the target sample size was reached. The median size of injecting networks and the homophily (a measure of in-group affiliation) of different subgroups were calculated. Participants’ information was linked with that of their recruiter to form dyads, and multivariate analysis was conducted to determine whether dyad and injecting network characteristics were associated with sharing injecting equipment within the dyads.ResultsThe injecting networks were large, the lowest median subgroup network size being 12. Homophily estimates indicated a lack of strong ties both within and across groups. In the multivariate analysis, factors significantly associated with sharing injecting equipment within dyads were feeling very close to their recruiter and having one or both members of the dyad identify as Aboriginal or Torres Strait Islander and one or both members having not been tested for hepatitis C in the previous year.ConclusionRDS provided valuable information on injecting networks in Sydney. PWID were shown to be socially connected with a large number of other injectors, and affiliations were formed without regard to demographic or drug use characteristics. Linking information from the recruits with that of their recruiter was a useful way of organizing information to gain a more complete understanding of risk behaviour