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The use of qualitative audio diaries in research allows people to generate multi-occasion reports of events in their natural, spontaneous contexts in a recorded audio format. Recent technological advances have enabled more rapid, spontaneous means for audio diaries recordings, such as smartphones. This article reports on methodological insights regarding the feasibility and contributions of using chatbots – digital systems that provide an interface for interaction with users that emulate human conversation – to assist the collection of audio diaries. We describe the development of a WhatsApp chatbot for the remote collection and sharing of audio files using a diary format. Methodological considerations are presented, drawing from the results of a remote collection of spoken reports from 19 Brazilian undergraduate students who were experiencing social isolation due to COVID-19 pandemic-related containment measures. Results emphasize the temporal aspect of the data collection process, and indicate that the tool allowed participants to generate accounts depicting changes in their understanding of perceptions and experiences over time. Although leveraging digital technology and conversational-like features of chatbots may promote greater participation and less participant burden, results also suggest that using pre-scripted prompts in the form of open questions may pose a challenge for reflexive engagement with audio diaries. [ABSTRACT FROM AUTHOR]

Centrosomes are the major microtubule organizing centers of animal cells. Supernumerary centrosomes are a common feature of human tumors and associated with karyotype abnormalities and aggressive disease, but whether they are cause or consequence of cancer remains controversial. Here, we analyzed the consequences of centrosome amplification by generating transgenic mice in which centrosome numbers can be increased by overexpression of the structural centrosome protein STIL. We show that STIL overexpression induces centrosome amplification and aneuploidy, leading to senescence, apoptosis, and impaired proliferation in mouse embryonic fibroblasts, and microcephaly with increased perinatal lethality and shortened lifespan in mice. Importantly, both overall tumor formation in mice with constitutive, global STIL overexpression and chemical skin carcinogenesis in animals with inducible, skin-specific STIL overexpression were reduced, an effect that was not rescued by concomitant interference with p53 function. These results suggest that supernumerary centrosomes impair proliferation in vitro as well as in vivo, resulting in reduced lifespan and delayed spontaneous as well as carcinogen-induced tumor formation. Author summary: Already at the beginning of the last century, Theodor Boveri suggested that supernumerary centrosomes might be a cause of cancer via the generation of abnormal mitotic spindles and subsequent chromosome missegregation. In the meantime, centrosome amplification has been observed in most tumor types and is viewed as a "hallmark" of cancer cells. However, studies on tumor formation in mice by overexpression of PLK4, the principal kinase regulating centrosome duplication, led to ambiguous results, as some but not all models resulted in increased tumorigenesis. In this study, we generated transgenic mice, which overexpress the structural centrosome protein STIL. Similar to PLK4, overexpression of STIL caused centrosome amplification and aneuploidy in mouse cells. Our analyses revealed that powerful mechanisms lead to the elimination of cells with extra centrosomes and/or aneuploidy by impaired proliferation, senescence and apoptosis, thereby delaying both spontaneous tumor formation and chemical skin carcinogenesis, and explaining the reduced life span of STIL-transgenic mice. [ABSTRACT FROM AUTHOR]

Simple Summary: In recent decades, interest in the use of donkey milk for human nutrition has increased, since it may represent a natural substitute for cow's milk for children affected by milk protein allergies. The functional peculiarities of donkey milk are mainly linked to its casein content comparable to that of human milk. This study provides a thorough analysis of transcript isoforms generated by two αs2-casein-encoding genes (CSN1S2 I and CSN1S2 II) in donkeys and a detection of significant genetic diversity at both loci along with sequence comparisons across species. In particular, a key mutation affecting exon 17 splicing in CSN1S2 I was identified, and a genotyping method was developed. These data represent an important step in the understanding of the expression regulation of these genes in donkeys and a useful tool for the genetic improvement of donkey milk production that fulfils special consumer requirements. The αs2-casein is a phosphoprotein secreted in the milk of most mammals, and it is the most hydrophilic of all caseins. Contrary to genes found in ruminants, in donkeys two different encoding genes for donkey αs2-casein (CSN1S2 I and CSN1S2 II) have been identified. However, unlike in ruminants, the variability at these loci has not been characterized in detail in donkeys until now. In this study, we analyze the transcript profile of the donkey CSN1S2 I and CSN1S2 II genes, and we identify and describe the variability of these loci in the Ragusana and Amiatina breeds reared in Italy. The analysis of the CSN1S2 I Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) products and subsequent sequencing showed, in addition to correctly spliced mRNA, seven other minor mRNAs resulting from differential splicing events involving, in various combinations, entire exons (4, 5, 6, and 11), parts of exons (5′ or 3′ end of exon 17), or the recognition of intronic sequences as an exon (exon 12′). Similarly, the transcription analysis of the CSN1S2 II gene revealed a remarkable variability in splicing events, mainly concerning the alternative insertion of an extra exon 7 (named 7′); the first 33 bp of exon 13; or the alternative skipping of exons 9, 10, 11, 12, and 15, and their combinations. At the mRNA level for CSN1S2 I, seven SNPs were observed, five of which led to amino acid changes: p.T73>A, p.I109>V, p.I130>V, p.I146>T, and p.D217>Y. Similarly, nine SNPs were observed at the CSN1S2 II locus, seven of which are non-synonymous: p.L63>F, p.H70>Q, p.D90>N, p.129A>T, p.H131>Y, p.E144>G, and p.F157>S. In addition, the DNA sequencing of exon 17 and flanking introns of the CSN1S2 I gene revealed a G>A transition at the splice acceptor site of CSN1S2 I exon 17 (FM946022.1:c.375-1G>A), resulting in an allele-specific skipping of the first 15 nucleotides of this exon, which encode the peptide 176NKINQ180, and the recognition of an in-frame cryptic splicing acceptor site: arAACAAAATCAACCAG. A genotyping method based on restriction fragment length polymorphism (XbaI PCR-RFLP) was set up for this SNP. In the total population studied (105 Ragusana and 14 Amiatina donkeys), the A allele had a frequency of 0.2437 with no evidence of deviation from the Hardy–Weinberg equilibrium. This study adds new knowledge regarding the genetic variability of αs2-caseins in donkeys and may contribute significantly to the genetic improvement of milk production for this species. [ABSTRACT FROM AUTHOR]

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Stroke survivors commonly face challenges such as reduced physical activity and cardiorespiratory fitness (CRF) as well as balance and gait impairments, exacerbating their disability. While high-intensity exercise interventions have demonstrated some potential, their effects on these items remain uncertain. Therefore, our study aimed to investigate the impact of high-intensity training protocols on CRF, gait ability, and balance in stroke survivor populations. Two independent investigators systematically searched five databases for relevant RCTs following the PICO model. Through a systematic review of 25 RCTs published up to 31 May 2023, including adult first-stroke survivors, comparing high-intensity exercise training versus low-to-mild or no exercises, we evaluated outcomes such as the Six-Minute Walking Test (6 MWT), peak oxygen uptake (VO2peak), Ten-Meter Walk Test (10 MWT), Berg Balance Scale (BBS), and Timed Up and Go test (TUG). The protocol was registered in PROSPERO (registration number CRD42023456773). Meta-analyses indicated significant enhancements in CRF, as measured by 6 MWT and VO2peak, following high-intensity exercise interventions. However, no significant differences were observed in BBS, 10 MWT, and TUG. Our findings underscore the potential of high-intensity exercise interventions in ameliorating CRF among stroke survivors, although further research involving standardized protocols and long-term follow-ups is imperative to optimize rehabilitation outcomes. [ABSTRACT FROM AUTHOR]

What subject lines and messages in an e-mailed invitation maximize cooperation for a web survey? Answering this question is ever more important as survey researchers increasingly turn to web surveys to reduce costs. We test respondents' preferences for subject lines and messages in an e-mailed invitation to participate in a Consumer Assessment of Healthcare Providers and Systems survey. Findings suggest that respondents prefer informative and straightforward subject lines that reference the purpose and the sponsor of the survey. Brief and uninformative subject lines should be avoided. In the body of the e-mail, respondents prefer messages that expand on the study's purpose and usefulness and extend a guarantee of confidentiality. Messages that encourage participation for the sake of the study's representativeness or accuracy are unpersuasive to potential respondents. Few differences emerged in preferences among demographic subgroups.

ObjectivesTo explore experiences of U.S. (United States) nursing home leadership during the COVID-19 pandemic in their efforts to address resident loneliness and social isolation and to elicit stories about personal and professional impacts on themselves and staff.DesignQualitative inquiry via three optional open-ended questions appended to a national self-administered survey of American nursing home leaders was employed. Textual data was analyzed using an iterative reflexive thematic approach.Setting and participantsA stratified sample frame defined by facility size (beds: 30-99, 100+) and quality ratings (1, 2-4, 5) was employed. Web survey links and paper surveys were sent to 1,676 nursing home directors of nursing between February and May 2022.ResultsOpen text responses were collected from 271 nursing homes. Broad themes included: 1) Addressing needs of residents & families; 2) Challenges; and 3) Personal experiences of nursing home leadership/staff. Respondents described trauma to residents, staff, and leadership. Resident loneliness was addressed using existing and newer technologies and innovative indoor and outdoor activities. Residents experienced fear, illness, loss, and sometimes death. Isolation from family and lack of touch were particularly difficult. Regulations were seen as punitive while ignoring emotional needs of residents. Staffing challenges and pressures to do more with less created additional stress. Leadership and staff made significant sacrifices resulting in physical, social, and emotional consequences. Beneficial outcomes included staff bonding, professional growth, and permanent implementation of new interventions.Conclusions and implicationsNew and creative interventions were successfully implemented to address social isolation and loneliness. Improved Wi-Fi and other nursing home infrastructure upgrades are needed to maintain them. Reimagining often conflicting overlapping federal, state, and local regulations, grounding them in good clinical judgement, and incentivizing performance improvement should be considered. Trauma experienced by staff needs to be addressed to deal with current and future workforce needs.

Cow's milk allergy (CMA) is one of the earliest and most common food allergy and can be elicited by both IgE- or non-IgE-mediated mechanism. We previously described dysbiosis in children with IgE-mediated CMA and the effect of dietary treatment with extensively hydrolyzed casein formula (EHCF) alone or in combination with the probiotic Lactobacillus rhamnosus GG (LGG). On the contrary, the gut microbiota in non-IgE-mediated CMA remains uncharacterized. In this study we evaluated gut microbiota composition and fecal butyrate levels in children affected by non-IgE-mediated CMA. We found a gut microbiota dysbiosis in non-IgE-mediated CMA, driven by an enrichment of Bacteroides and Alistipes. Comparing these results with those previously obtained in children with IgE-mediated CMA, we demonstrated overlapping signatures in the gut microbiota dysbiosis of non-IgE-mediated and IgE-mediated CMA children, characterized by a progressive increase in Bacteroides from healthy to IgE-mediated CMA patients. EHCF containg LGG was more strongly associated with an effect on dysbiosis and on butyrate production if compared to what observed in children treated with EHCF alone. If longitudinal cohort studies in children with CMA will confirm these results, gut microbiota dysbiosis could be a relevant target for innovative therapeutic strategies in children with non-IgE-mediated CMA.