Functional Enhancement of Electrofusion-derived BRIN-BD11 Insulin-secreting Cells After Implantation into Diabetic Mice

Electrofusion-derived BRIN-BD11 cells are glucosesensitive insulin-secreting cells which provide an archetypal bioengineered surrogate β-cell for insulin replacement therapy in diabetes mellitus, 5x106 BRIN-BD11 cells were implanted intraperitoneally into severely hyperglycaemic (>24mmol/l) streptozotocin-induced insulin-treated diabetic athymic nude (nu/nu) mice. The implants reduced hyperglycaemia such that insulin injections were discontinued by 5–16 days (<17mmol/l) and normoglycaemia (<9mmol/l) was achieved by 7–20 days. Implanted cells were removed after 28 days and re-established in culture. After re-culture for 20 days, glucose-stimulated (16.7mmol/l) insulin release was enhanced by 121% (p<0.001) compared to non-implanted cells. Insulin responses to glucagon-like peptide-1 (10−9mol/l), cholecystokinin-8 (10−8 mol/l) and L-alanine (10 mmol/l) were increased by 32%, 31% and 68% respectively (p<0.05–0.01). Insulin content of the cells was 148% greater at 20 days after re-culture than before implantation (p<0.001), but basal insulin release (at 5.6 mmol/l glucose) was not changed. After re-culture for 40 days, insulin content declined to 68% of the content before implantation (p<0.01), although basal insulin release was unchanged. However, the insulin secretory responses to glucose, glucagonlike peptide-1, cholecystokinin-8 and L-alanine were decreased after 40 days of re-culture to 65%, 72%, 73% and 42% respectively of the values before implantation (p<0.05–0.01). The functional enhancement of electrofusion-derived surrogate β-cells that were re-cultured for 20 days after implantation and restoration of normoglycaemia indicates that the in vivo environment could greatly assist β-cell engineering approaches to therapy for diabetes

Change of direction in the biomechanics of atherosclerosis

The non-uniform distribution of atherosclerosis within the arterial system has been attributed to pro-atherogenic influences of low, oscillatory haemodynamic wall shear stress (WSS) on endothelial cells (EC). This theory is challenged by the changes in lesion location that occur with age in human and rabbit aortas. Furthermore, a number of point-wise comparisons of lesion prevalence and WSS have failed to support it. Here we investigate the hypothesis that multidirectional flow-characterized as the average magnitude of WSS components acting transversely to the mean vector (transWSS)-plays a key role. Maps of lesion prevalence around aortic branch ostia in immature and mature rabbits were compared with equivalent maps of time average WSS, the OSI (an index characterizing oscillatory flow) and transWSS, obtained from computational simulations; Spearman's rank correlation coefficients were calculated for aggregated data and 95% confidence intervals were obtained by bootstrapping methods. Lesion prevalence correlated positively, strongly and significantly with transWSS at both ages. Correlations of lesion prevalence with the other shear metrics were not significant or were significantly lower than those obtained for transWSS. No correlation supported the low, oscillatory WSS theory. The data are consistent with the view that multidirectional near-wall flow is highly pro-atherogenic. Effects of multidirectional flow on EC, and methods for investigating them, are reviewed. The finding that oscillatory flow has pro-inflammatory effects when acting perpendicularly to the long axis of EC but anti-inflammatory effects when acting parallel to it may explain the stronger correlation of lesion prevalence with transWSS than with the OSI

Estimating arterial cyclic strain from the spacing of endothelial nuclei

Background: The non-uniform distribution of atherosclerosis within the arterial system is widely attributed to variation in haemodynamic wall shear stress. It may also depend on variation in pressure-induced stresses and strains within the arterial wall; these have been less widely investigated, at least in part because of a lack of suitable techniques. Objectives: Here we show that local arterial strain can be determined from impressions left by endothelial cells on the surface of vascular corrosion casts made at different pressures, even though only one pressure can be examined in each vessel. The pattern of pits in the cast caused by protruding endothelial nuclei was subject to “retro-deformation” to identify the pattern that would have occurred in the absence of applied stresses. Methods: Retaining the nearest-neighbour pairs found under this condition, changes in nearest-neighbour vectors were calculated for the pattern seen in the cast, and the ratio of mean changes at different pressures determined. This approach removes errors in simple nearest-neighbour analyses caused by the nearest neighbour changing as deformation occurs. Results: The accuracy, precision and robustness of the approach were validated using simulations. The method was implemented using confocal microscopy of casts of the rabbit aorta made at systolic and diastolic pressures; results agreed well with the ratio of the macroscopic dimensions of the casts. Conclusions: Applying the new technique to areas around arterial branches could support or refute the hypothesis that the development of atherosclerosis is influenced by mural strain, and the method may be applicable to other tissues

“It just makes me feel a little less alone” a qualitative exploration of the podcast “menopause unmuted” on women's perceptions of menopause

Objective: Menopause can negatively impact women's quality of life, with many women reporting inadequate information and support. Podcasts have grown in popularity in recent years and have been found to be accessible methods for increasing knowledge and challenging perceptions of stigmatized topics. The current research aimed to understand the impact of the podcast “menopause: unmuted” on women's menopause-related knowledge, understanding, and communication practices. Methods: A diverse sample of 30 women aged 40 to 60 years listened to the podcast series, which focused on menopause stories, before taking part in semistructured interviews to discuss the impact of the podcast on how they understood and communicated about menopause. The interviews were analyzed thematically. Results: Two overarching themes were identified in the data. A “journey of knowledge gain” explores participants’ understanding of menopause before listening to the podcast and describes how this is deepened by hearing and connecting with women's stories. “Reframing menopause” describes the impact of the podcast, where women reflect on the value of communication amongst women, challenge and re-evaluate the stigmatization of menopause, and discuss ways to make positive behavioral changes in their lives. Conclusions: The podcast “menopause: unmuted” helped women to learn about the menopause experience, have a greater sense of belonging to a community of women, and feel empowered to make changes in their own lives. Sharing stories via podcasts has potential as an accessible and impactful medium to educate women and reduce the widespread stigma associated with menopause

"You're more engaged when you're listening to somebody tell their story": A qualitative exploration into the mechanisms of the podcast 'menopause: unmuted' for communicating health information.

While health podcasts can be effective in reducing stigma and increasing knowledge, we know little about their mechanisms of action. This qualitative study explored the mechanisms of how women connected with the podcast 'menopause: unmuted', which presented menopause information in a storytelling format. A diverse sample of 30 women aged 40-60 years were interviewed after listening to the podcast. Interviews covered participant's views and perceptions of the stories presented. Transcripts were analyzed thematically. Two overarching themes were identified. 'Openness and authenticity' describes the value of personal stories told in an authentic way by real experts-by-experience. 'Relatability and representation' explores participants' emotional reaction to the podcast, influenced by the extent to which they identified with the stories and storytellers on the podcast. Authenticity and relatability were identified as key mechanisms through which participants connected with audio stories, consistent with Fisher's narrative theory. These findings have important implications for the application of storytelling in podcasts designed to influence health behaviors. Diverse stories representing a range of demographic characteristics and experiences are needed when creating podcasts about health information to increase listener's relatability and connection. [Abstract copyright: Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.

Effects of dietary salt on gene and protein expression in brain tissue of a model of sporadic small vessel disease

Background: The effect of salt on cerebral small vessel disease (SVD) is poorly understood. We assessed the effect of dietary salt on the cerebral tissue of the strokeprone spontaneously hypertensive rat (SHRSP) - a relevant model of sporadic SVD - at both the gene and protein level. Methods: Brains from 21 week old SHRSP and Wistar-Kyoto rats, half additionally salt-loaded (via a 3 week regime of 1% NaCl in drinking water) were split into 2 hemispheres and sectioned coronally – one hemisphere for mRNA microarray and qRT-PCR, the other for immunohistochemistry using a panel of antibodies targeting components of the neurovascular unit. Results: We observed differences in gene and protein expression affecting the acute phase pathway and oxidative stress (ALB, AMBP, APOH, AHSG and LOC100129193, up-regulated in salt-loaded WKY versus WKY, &gt;2-fold), active microglia (increased Iba-1 protein expression in salt-loaded SHRSP versus saltloaded WKY, p&lt;0.05), vascular structure (ACTB &amp; CTNNB, up-regulated in saltloaded SHRSP versus SHRSP, &gt;3-fold; CLDN-11,VEGF and VGF downregulated &gt;- 2-fold in salt-loaded SHRSP versus SHRSP) and myelin integrity (MBP downregulated in salt loaded WKY rats versus WKY, &gt;2.5-fold). Changes of salt-loading were more pronounced in SHRSP and occurred without an increase in blood pressure in WKY rats. Conclusion: Salt exposure induced changes in gene and protein expression in an experimental model of SVD and its parent rat strain in multiple pathways involving components of the glio-vascular unit. Further studies in pertinent experimental models at different ages would help clarify the short and long-term effect of dietary salt in SVD

Mass drug administration with azithromycin for trachoma elimination and the population structure of Streptococcus pneumoniae in the nasopharynx

ABSTRACTBackgroundMass drug administration (MDA) with azithromycin for trachoma elimination reduces nasopharyngeal carriage of Streptococcus pneumoniae in the short term. We evaluated S. pneumoniae carried in the nasopharynx before and after a round of azithromycin MDA to determine whether MDA was associated with changes in pneumococcal population structure.MethodsWe analysed 514 pneumococcal isolates cultured from nasopharyngeal samples collected in Gambian villages that received MDA for trachoma elimination. The samples were collected during three cross-sectional surveys conducted before the third round of MDA (CSS-1) and at one (CSS-2) and six (CSS-3) months after MDA. Whole genome sequencing was conducted on randomly selected isolates. Bayesian Analysis of Population Structure (BAPS) was used to cluster related isolates by capturing variation in the core genome. Serotype and multi-locus sequence type were inferred from the genotype. The Antimicrobial Resistance Identification by Assembly (ARIBA) tool was used to identify macrolide resistance genes.ResultsTwenty-seven BAPS clusters were assigned. These consisted of 81 sequence types (STs), 15 of which were novel additions to pubMLST. Two BAPS clusters, BAPS20 (p-value&lt;=0.016) and BAPS22 (p-value&lt;=0.032) showed an increase in frequency at CSS-3 not associated with antimicrobial resistance. Macrolide resistance within BASP17 increased after treatment (p&lt;0.05) and was carried on a mobile transposable element that also conferred resistance to tetracycline.ConclusionsLimited changes in pneumococcal population structure were observed after the third round of MDA suggesting treatment had little effect on the circulating lineages. An increase in macrolide resistance within one BAPS highlights the need for antimicrobial resistance surveillance in treated villages.</jats:sec

Mass administration of azithromycin and Streptococcus pneumoniae carriage: cross-sectional surveys in the Gambia.

OBJECTIVE: To evaluate the effect of repeated mass drug administration (MDA) of azithromycin in the Gambia on the nasopharyngeal carriage of Streptococcus pneumoniae and on the emergence of antibiotic-resistant strains. METHODS: This study involved villages that participated in a cluster randomized trial comparing the effect of one versus three azithromycin MDA rounds on the prevalence of trachoma. Only villages in which most children received 7-valent pneumococcal conjugate vaccine were included. Three cross-sectional surveys were performed in two villages that received three annual MDA rounds: the first immediately before the third MDA round and the second and third, 1 and 6 months, respectively, after the third MDA round. The third survey also covered six villages that had received one MDA round 30 months previously. Pneumococcal carriage was assessed using nasopharyngeal swabs and azithromycin resistance was detected using the Etest. FINDINGS: The prevalence of pneumococcal carriage decreased from 43.4% to 19.2% between the first and second surveys (P < 0.001) but rebounded by the third survey (45.8%; P = 0.591). Being a carrier at the first survey was a risk factor for being a carrier at the second (odds ratio: 3.71; P <  0.001). At the third survey, the prevalence of carriage was similar after one and three MDA rounds (50.3% versus 45.8%, respectively; P = 0.170), as was the prevalence of azithromycin resistance (0.3% versus 0.9%, respectively; P = 0.340). CONCLUSION: Three azithromycin MDA rounds did not increase the prevalence of nasopharyngeal carriage of azithromycin-resistant S. pneumoniae strains compared with one round

Jaw claudication and jaw stiffness in giant cell arteritis: Secondary analysis of a qualitative research dataset

Objectives Jaw symptoms can be a vital clue to the diagnosis of giant cell arteritis (GCA). Guidelines recommend more intensive treatment if jaw claudication is present. We sought to explore how patients with GCA described their jaw symptoms. Methods Secondary, qualitative analysis of interview data from 36 participants from the UK (n = 25) and Australia (n = 11), originally collected in order to develop a patient-reported outcome measure for GCA. In all cases, GCA had been confirmed by biopsy/imaging. Interview transcripts were organised within QSR NVivo 12 software, and analysed using template analysis. Themes were refined through discussion among the research team including a patient partner. Results 20/36 participants reported jaw symptoms associated with GCA. Median age of these 20 participants was 76.5 years; 60% were female. Five themes were identified: physical sensations; impact on function; impact on diet; symptom response with steroids; attribution to other causes. Physical sensations included ache, cramp, stiffness and “lock-jaw”. Functional impacts included difficulty in eating/chewing, cleaning teeth, speaking, or opening the mouth. Dietary impacts included switching to softer food. Response to steroids was not always immediate. Jaw symptoms were initially mis-attributed by some participants to arthritis, age or viral illnesses; or by healthcare professionals to dental cavity, ear infection or teeth-grinding. Conclusion Jaw symptoms in GCA are diverse and can lead to diagnostic confusion with primary temporomandibular joint (TMJ) disorder, potentially contributing to delay in GCA diagnosis. Further research is needed to determine the relationship of jaw stiffness to jaw claudication. Lay Summary Giant cell arteritis (GCA) causes inflammation of some of the larger blood vessels of the body, especially around the head and shoulders. If not treated, GCA can cause sight loss. Therefore, prompt diagnosis is important. Doctors are taught that one of the vital clues to GCA is jaw claudication: pain that comes on with chewing and resolves with rest. Guidelines state that patients presenting with jaw claudication need more intensive treatment. We looked back at interviews that had been done for a study to develop a questionnaire about the impact of GCA on patients. In these interviews, over half the participants described experiencing jaw symptoms, but these symptoms were not always typical “jaw claudication”. Jaw stiffness or difficulty opening the mouth were also described. For some patients, difficulty opening the mouth had the greatest impact on diet. Clinicians should be aware that GCA can present with a variety of jaw symptoms. Further research is needed to define the symptom of jaw claudication more precisely, as it may have diagnostic and treatment implications