The dynamic emergence of GATA1 complexes identified in in vitro embryonic stem cell differentiation and in vivo mouse fetal liver.

GATA1 is an essential transcriptional regulator of myeloid hematopoietic differentiation towards red blood cells. During erythroid differentiation, GATA1 forms different complexes with other transcription factors such as LDB1, TAL1, E2A and LMO2 ("the LDB1 complex") or with FOG1. The functions of GATA1 complexes have been studied extensively in definitive erythroid differentiation; however, the temporal and spatial formation of these complexes during erythroid development is unknown. We applied proximity ligation assay (PLA) to detect, localize and quantify individual interactions during embryonic stem cell differentiation and in mouse fetal liver (FL) tissue. We show that GATA1/LDB1 interactions appear before the proerythroblast stage and increase in a subset of the CD71+/TER119- cells to activate the terminal erythroid differentiation program in 12.5 day FL. Using Ldb1 and Gata1 knockdown FL cells, we studied the functional contribution of the GATA1/LDB1 complex during differentiation. This shows that the active LDB1 complex appears quite late at the proerythroblast stage of differentiation and confirms the power of PLA in studying the dynamic interaction of proteins in cell differentiation at the single cell level. We provide dynamic insight into the temporal and spatial formation of the GATA1 and LDB1 transcription factor complexes during hematopoietic development and differentiation.

Phonetic change in an Antarctic winter.

An acoustic analysis was made of the speech characteristics of individuals recorded before and during a prolonged stay in Antarctica. A computational model was used to predict the expected changes due to close contact and isolation, which were then compared with the actual recorded productions. The individuals were found to develop the first stages of a common accent in Antarctica whose phonetic characteristics were in some respects predicted by the computational model. These findings suggest that the phonetic attributes of a spoken accent in its initial stages emerge through interactions between individuals causing speech production to be incrementally updated.

The genome-wide dynamics of the binding of Ldb1 complexes during erythroid differentiation.

One of the complexes formed by the hematopoietic transcription factor Gata1 is a complex with the Ldb1 (LIM domain-binding protein 1) and Tal1 proteins. It is known to be important for the development and differentiation of the erythroid cell lineage and is thought to be implicated in long-range interactions. Here, the dynamics of the composition of the complex-in particular, the binding of the negative regulators Eto2 and Mtgr1-are studied, in the context of their genome-wide targets. This shows that the complex acts almost exclusively as an activator, binding a very specific combination of sequences, with a positioning relative to transcription start site, depending on the type of the core promoter. The activation is accompanied by a net decrease in the relative binding of Eto2 and Mtgr1. A Chromosome Conformation Capture sequencing (3C-seq) assay also shows that the binding of the Ldb1 complex marks genomic interaction sites in vivo. This establishes the Ldb1 complex as a positive regulator of the final steps of erythroid differentiation that acts through the shedding of negative regulators and the active interaction between regulatory sequences.

[Oligoarthritis, panniculitis and pyoderma gangrenosum associated with non-Hodgkin lymphoma. Report of one case]. / Oligoartritis, paniculitis y pioderma gangrenoso como manifestaciones de un Linfoma no Hodgkin.

An assortment of clinical and laboratory abnormalities may occur as paraneoplastic syndromes in lymphomas. Rheumatological and dermatological manifestations such as paraneoplastic arthritis and pyoderma gangrenosum must be underscored. We report a 28 years old woman who developed pyoderma gangrenosum and two years later presented with arthritis of knees and ankles associated with panniculitis interpreted as erythema induratum that was pathologically confirmed. She developed a reactivation of pyoderma gangrenosum, that was refractory to treatment. Complementary studies showed a pulmonary nodule and a right paravertebral mass with involvement of the psoas muscle. Biopsies of both masses and a new pathological skin study demonstrated a large B-cell non-Hodgkin's lymphoma.

Dynamic long-range chromatin interactions control Myb proto-oncogene transcription during erythroid development.

The key haematopoietic regulator Myb is essential for coordinating proliferation and differentiation. ChIP-Sequencing and Chromosome Conformation Capture (3C)-Sequencing were used to characterize the structural and protein-binding dynamics of the Myb locus during erythroid differentiation. In proliferating cells expressing Myb, enhancers within the Myb-Hbs1l intergenic region were shown to form an active chromatin hub (ACH) containing the Myb promoter and first intron. This first intron was found to harbour the transition site from transcription initiation to elongation, which takes place around a conserved CTCF site. Upon erythroid differentiation, Myb expression is downregulated and the ACH destabilized. We propose a model for Myb activation by distal enhancers dynamically bound by KLF1 and the GATA1/TAL1/LDB1 complex, which primarily function as a transcription elongation element through chromatin looping.

X chromosome inactivation is initiated in human preimplantation embryos.

X chromosome inactivation (XCI) is the mammalian mechanism that compensates for the difference in gene dosage between XX females and XY males. Genetic and epigenetic regulatory mechanisms induce transcriptional silencing of one X chromosome in female cells. In mouse embryos, XCI is initiated at the preimplantation stage following early whole-genome activation. It is widely thought that human embryos do not employ XCI prior to implantation. Here, we show that female preimplantation embryos have a progressive accumulation of XIST RNA on one of the two X chromosomes, starting around the 8-cell stage. XIST RNA accumulates at the morula and blastocyst stages and is associated with transcriptional silencing of the XIST-coated chromosomal region. These findings indicate that XCI is initiated in female human preimplantation-stage embryos and suggest that preimplantation dosage compensation is evolutionarily conserved in placental mammals.

Mitigation of SARS-CoV-2 transmission at a large public university.

In Fall 2020, universities saw extensive transmission of SARS-CoV-2 among their populations, threatening health of the university and surrounding communities, and viability of in-person instruction. Here we report a case study at the University of Illinois at Urbana-Champaign, where a multimodal "SHIELD: Target, Test, and Tell" program, with other non-pharmaceutical interventions, was employed to keep classrooms and laboratories open. The program included epidemiological modeling and surveillance, fast/frequent testing using a novel low-cost and scalable saliva-based RT-qPCR assay for SARS-CoV-2 that bypasses RNA extraction, called covidSHIELD, and digital tools for communication and compliance. In Fall 2020, we performed >1,000,000 covidSHIELD tests, positivity rates remained low, we had zero COVID-19-related hospitalizations or deaths amongst our university community, and mortality in the surrounding Champaign County was reduced more than 4-fold relative to expected. This case study shows that fast/frequent testing and other interventions mitigated transmission of SARS-CoV-2 at a large public university.

Perioperative chemotherapy in locally advanced gastric cancer in Chile: from evidence to daily practice.

Gastric cancer (GC) is a leading cause of cancer death in Chile. Although recommended in international guidelines since 2006, perioperative chemotherapy was not available to patients in the public health system in Chile until 2016. We conducted an observational study to assess the feasibility of this strategy in public hospitals in Chile (Observational Study of Perioperative Chemotherapy in Locally Advanced Gastric Cancer - PRECISO). Patients with locally advanced, operable GC were offered to receive preoperative chemotherapy with Epirubicin + Cisplatin + Capecitabine (ECX) for three cycles followed by curative surgery. Staging included abdominal CT scan and laparoscopy if peritoneal carcinomatosis was suspected. Postoperative ECX for three cycles was recommended. Between August 2010 and March 2013, 110 patients were screened and 61 enrolled. Median age was 62 years (23-76 years) and most patients had good performance status at baseline (Eastern Cooperative Oncology Group performance status score (ECOG) 0: 42, ECOG 1: 19). Tumour site was proximal in 32 (52%) and medial and distal in 29 (48%) patients. All but four patients (n = 57, 93%) completed three cycles of preoperative chemotherapy. Fifty-six patients were operated and 54 (89%) had a curative resection. Thirty-three patients (54%) had pT0-2, and 18 (30%) had pN0 tumours, with two patients achieving a complete response. As of 20 December 2020, 39 patients died, 32 due to GC, one within 30 days of surgery, two due to intestinal obstruction at 5 and 3 months after surgery and four due to other causes. Five-year survival rate was 38%. We conclude that perioperative chemotherapy is feasible in public hospitals in Chile and should be offered to patients with locally advanced GC.

7-Tesla MRI of the brain in a research subject with bilateral, total knee replacement implants: Case report and proposed safety guidelines.

Recently, the first 7-T MR system was approved for clinical use in the United States. Unfortunately, relatively few metallic implants have undergone testing to determine if they are acceptable or pose hazards to research subjects and patients at this ultra-high-field strength. Therefore, in lieu of not performing a research or clinical MRI exam at 7-T, the supervising physician may make a decision to scan the individual with an untested metallic implant based on an analysis of the risks vs. the benefits. We present a case report of a research subject with bilateral, total knee replacement implants that safely underwent MRI of the brain at 7-T and provide guidelines for healthcare professionals to follow in order to ensure safety in research subjects or patients with metallic implants referred for 7-T scans.

Long-term risk of fatal malignancy following pediatric radiofrequency ablation.

Children undergoing radiofrequency ablation (RFA) are believed to be at increased risk of developing malignancy caused by radiation, although the magnitude of this risk is incompletely understood. We previously reported a strategy to reduce radiation exposure during pediatric RFA. In a cohort of 15 subjects (median age 12 years, range 9 to 17), radiation was measured using dosimeters at 5 sites. The risk of malignancy using measured radiation absorbed dose was calculated. International Council for Radiation Protection 60 risk estimates were applied to calculate absorbed organ doses. Median duration of combined biplane fluoroscopy was 14.4 minutes. Of the 5 dosimeter locations, the right scapular location had the highest median radiation exposure (43 mGy). Incorporating data from the 5 dosimeters, the risk model calculated that the organ with the greatest absorbed dose and at greatest risk of malignancy was the lung, followed by bone marrow, then breast. Thyroid and ovary exposures were negligible. The increased lifetime risk of fatal malignancy was 0.02% per single RFA procedure. In conclusion, with appropriate measures to reduce radiation exposure, the increased risk of malignancy after a single RFA procedure in children is low. These data should be of help counseling families and will contribute to analysis of the relative risk reduction benefits of such novel imaging approaches as a magnetic resonance imaging-based catheterization laboratory.

Linking Cognitive and Social Aspects of Sound Change Using Agent-Based Modeling.

The paper defines the core components of an interactive-phonetic (IP) sound change model. The starting point for the IP-model is that a phonological category is often skewed phonetically in a certain direction by the production and perception of speech. A prediction of the model is that sound change is likely to come about as a result of perceiving phonetic variants in the direction of the skew and at the probabilistic edge of the listener's phonological category. The results of agent-based computational simulations applied to the sound change in progress, /u/-fronting in Standard Southern British, were consistent with this hypothesis. The model was extended to sound changes involving splits and mergers by using the interaction between the agents to drive the phonological reclassification of perceived speech signals. The simulations showed no evidence of any acoustic change when this extended model was applied to Australian English data in which /s/ has been shown to retract due to coarticulation in /str/ clusters. Some agents nevertheless varied in their phonological categorizations during interaction between /str/ and /ʃtr/: This vacillation may represent the potential for sound change to occur. The general conclusion is that many types of sound change are the outcome of how phonetic distributions are oriented with respect to each other, their association to phonological classes, and how these types of information vary between speakers that happen to interact with each other.

Using a virtual care scenario to enhance student learning.

A series of care scenarios were developed entitled 'eWARD' that complement traditional teaching methods employed in the education of nursing students. This article discusses the development of one of these scenarios, which was based on a stroke rehabilitation theme, in relation to its theoretical underpinnings as well as its implementation and evaluation.

Restless Legs Syndrome/Willis-Ekbom Disease Morbidity: Burden, Quality of Life, Cardiovascular Aspects, and Sleep.

Restless legs syndrome (RLS)/Willis-Ekbom disease (WED) has a significant negative effect on quality of life. The decreased quality of life is similar to that of other chronic diseases, such as diabetes type 2, depression, and osteoarthritis. RLS/WED disrupts sleep length, sleep quality, and daytime alertness. Sleep disruption can contribute to depression. RLS/WED has been associated with cardiovascular disease and high blood pressure, possibly because of increased sympathetic tone caused by periodic limb movements of sleep. RLS/WED is underdiagnosed, leading to chronic sleep disruption and daytime consequences. Patients with RLS/WED have decreased productivity at work, which potentially has far-reaching economic consequences.

Fat-sugar see-saw in school lunches: impact of a low fat intervention.

PURPOSE: To determine the long-term effects of a low fat intervention on sugar content in school lunches METHODS: We calculated contributions of total sugars, 6 specific sugars, and selected nutrients in National School Lunch Program meals served in 56 former intervention, 20 randomly selected control schools that had participated 3 years before in a low fat intervention, and 12 additional schools from neighboring school districts that had never been exposed to the intervention, whose goal was the lowering of total fat to under 30% of calories and saturated fat to under 10% calories. Analysis of variance adjusted for region was used to compare treatment groups. Pearson partial correlations controlling for the effects of region and treatment group were used to assess the strength of sugar and fat relationships. RESULTS: Three years after the low fat intervention, former intervention, control and unexposed schools lunches were similar in mean total sugars (25% of calories), and "added" sugars (e.g. sucrose, glucose, galactose, and maltose) at 15% of calories; and differed only in their lactose content. As percent of calories from fat or saturated fat in lunches decreased, that from sugars increased. Lunches that met reduced saturated fat goals were significantly higher than those not meeting goals in percent of calories from sugars both in meals as offered (27.6 +/- 0.3% vs. 26.2 +/- 0.3 SE p =.004) and as served (26.5 +/- 0.4 vs. 23.9 +/- 0.4 p =.009). Lunches meeting reduced total fat goals were significantly higher only in percent of calories from sugars as served. Seventy-five percent of total sugar in lunches offered was from chocolate milk, fruit/fruit juices, and white milk. More "added" sugar came from high micronutrient foods, such as fruit, fruit juices, and chocolate milk than from desserts and entrees. CONCLUSIONS: The existence of a fat-sugar "see-saw" makes it important to emphasize substitutions of fat and saturated fat with starches and fiber in school lunches.

A distal region of the human TGM1 promoter is required for expression in transgenic mice and cultured keratinocytes.

BACKGROUND: TGM1(transglutaminase 1) is an enzyme that crosslinks the cornified envelope of mature keratinocytes. Appropriate expression of the TGM1 gene is crucial for proper keratinocyte function as inactivating mutations lead to the debilitating skin disease, lamellar ichthyosis. TGM1 is also expressed in squamous metaplasia, a consequence in some epithelia of vitamin A deficiency or toxic insult that can lead to neoplasia. An understanding of the regulation of this gene in normal and abnormal differentiation states may contribute to better disease diagnosis and treatment. METHODS: In vivo requirements for expression of the TGM1 gene were studied by fusing various lengths of promoter DNA to a reporter and injecting the DNA into mouse embryos to generate transgenic animals. Expression of the reporter was ascertained by Western blotting and immunohistochemistry. Further delineation of a transcriptionally important distal region was determined by transfections of progressively shortened or mutated promoter DNA into cultured keratinocytes. RESULTS: In vivo analysis of a reporter transgene driven by the TGM1 promoter revealed that 1.6 kilobases, but not 1.1 kilobases, of DNA was sufficient to confer tissue-specific and cell layer-specific expression. This same region was responsible for reporter expression in tissues undergoing squamous metaplasia as a response to vitamin A deprivation. Mutation of a distal promoter AP1 site or proximal promoter CRE site, both identified as important transcriptional elements in transfection assays, did not prevent appropriate expression. Further searching for transcriptional elements using electrophoretic mobility shift (EMSA) and transfection assays in cultured keratinocytes identified two Sp1 elements in a transcriptionally active region between -1.6 and -1.4 kilobases. While mutation of either Sp1 site or the AP1 site singly had only a small effect, mutation of all three sites eliminated nearly all the transcriptional activity. CONCLUSIONS: A distal region of the TGM1 gene promoter, containing AP1 and Sp1 binding sites, is evolutionarily conserved and responsible for high level expression in transgenic mice and in transfected keratinocyte cultures.

Legal requirements for safe handling in community care.

CASE STUDY: Andrea, a district nurse, together with home carers from the local social services department, has been caring for a wheelchair-bound patient with multiple sclerosis for 3 years. The patient requires assistance with going to the toilet and going to bed. She has always been reluctant to be moved using a hoist because of sudden spasms and this causes her great anxiety. Now the patient is refusing to be move mechanically and will only consent to being lifted manually. Andrea and the home care staff are concerned that this might place them at risk of injury and argue that it is against the trust and local authority moving and handling policy, which has a no-lift clause. Andrea would like to refuse to manually lift the patient but decides to check the legal position first.

Manual handling and the lawfulness of no-lift policies.

Recent discussions over the plight of a woman who had spent a year sleeping in her wheelchair because the local NHS trust refused to allow her to be lifted into bed highlight the dilemma facing nurses when they are required to manually handle patients (Andrews and Robinson 2003). Nurses are acutely aware that the manual handling of patients can result in back injury (McGuire and Dewar 1993). Yet they are also aware that they owe a duty to patients to provide nursing care to meet their needs and that might only be achieved through manual handling. There is evidence that hospitals in particular have still not met the requirements of the Manual Handling Operations Regulations (Trevelyan 2000). The hazards of working in the community environment have generally been better addressed as part of the assessment for and delivery of individual care packages. This article considers the law's approach to balancing the opposing needs of nurses' and patients' health needs.

Oligoartritis, paniculitis y pioderma gangrenoso como manifestaciones de un Linfoma no Hodgkin / Oligoarthritis, panniculitis and pyoderma gangrenosum associated with non-Hodgkin lymphoma: report of one case

An assortment of clinical and laboratory abnormalities may occur as paraneoplastic syndromes in lymphomas. Rheumatological and dermatological manifestations such as paraneoplastic arthritis and pyoderma gangrenosum must be underscored. We report a 28 years old woman who developed pyoderma gangrenosum and two years later presented with arthritis of knees and ankles associated with panniculitis interpreted as erythema induratum that was pathologically confirmed. She developed a reactivation of pyoderma gangrenosum, that was refractory to treatment. Complementary studies showed a pulmonary nodule and a right paravertebral mass with involvement of the psoas muscle. Biopsies of both masses and a new pathological skin study demonstrated a large B-cell non-Hodgkin's lymphoma.

Systematic application of multiplex PCR enhances the detection of bacteria, parasites, and viruses in stool samples.

OBJECTIVES: To determine whether systematic testing of faecal samples with a broad range multiplex PCR increases the diagnostic yield in patients with diarrhoea compared with conventional methods and a clinician initiated testing strategy. METHODS: 1758 faecal samples from 1516 patients with diarrhoea submitted to two diagnostic laboratories were tested for viral, bacterial, and parasitic pathogens by Fast-Track Diagnostics multiplex real-time PCR kits and conventional diagnostic tests. RESULTS: Multiplex PCR detected pathogens in 530 samples (30%): adenovirus (51, 3%), astrovirus (95, 5%), norovirus (172, 10%), rotavirus (3, 0.2%), Campylobacter jejuni/coli (85, 5%), Salmonella spp. (22, 1%), Clostridium difficile (72, 4%), entero-haemorrhagic Escherichia coli (21, 1%), Cryptosporidium spp. (3, 0.2%), Entamoeba histolytica (1, 0.1%), and Giardia lamblia (59, 3%). In contrast, conventional testing detected a pathogen in 324 (18%) samples. CONCLUSIONS: Using a systematic approach to the diagnosis of gastroenteritis improved diagnostic yield. This enhanced detection with PCR was achieved by a combination of improved detection of individual pathogens and detection of pathogens not requested or unable to be tested by conventional tests. This approach also allowed earlier identification for most pathogens and created a workflow which is likely to adapt well for many diagnostic laboratories.

The medical educator, the discourse analyst, and the phonetician: a collaborative feedback methodology for clinical communication.

Frameworks for clinical communication assist educators in making explicit the principles of good communication and providing feedback to medical trainees. However, existing frameworks rarely take into account the roles of culture and language in communication, which can be important for international medical graduates (IMGs) whose first language is not English. This article describes the collaboration by a medical educator, a discourse analyst, and a phonetician to develop a communication and language feedback methodology to assist IMG trainees at a Victorian hospital in Australia with developing their doctor-patient communication skills. The Communication and Language Feedback (CaLF) methodology incorporates a written tool and video recording of role-plays of doctor-patient interactions in a classroom setting or in an objective structured clinical examination (OSCE) practice session with a simulated patient. IMG trainees receive verbal feedback from their hospital-based medical clinical educator, the simulated patient, and linguists. The CaLF tool was informed by a model of language in context, observation of IMG communication training, and process evaluation by IMG participants during January to August 2009. The authors provided participants with a feedback package containing their practice video (which included verbal feedback) and the completed CaLF tool.The CaLF methodology provides a tool for medical educators and language practitioners to work collaboratively with IMGs to enhance communication and language skills. The ongoing interdisciplinary collaboration also provides much-needed applied research opportunities in intercultural health communication, an area the authors believe cannot be adequately addressed from the perspective of one discipline alone.