Comparative study of factors influencing tension lap splices in reinforced concrete beams

The practice of splicing reinforcing bars in reinforced concrete structures to manage insufficient bar length is a common approach, which is mainly due to transportation limitations on bar length. The splicing of reinforcing bars side by side offers a simple and economical solution to the problem of continuity. This paper examines the influence of different structural parameters such as concrete cover, lap splice length, shear links confinement and concrete strength on the lap splices based on an extensive experimental database of laps and anchorage. The current study shows that increasing the lap splices beyond 50∅ has no additional benefit for increasing its strength. The results also show that relative to the measured stress, specimens with larger concrete side covers shows higher splice stress compared to the samples with smaller concrete covers

Stone axes throw new light on Baltic stone age mortuary rites

Despite their ubiquity, Mesolithic lithic tools given as funerary offerings have rarely been studied in detail. Whereas personal ornaments (e.g. beads, pendants) are commonly interpreted as markers of social identity and status, archaeologists have struggled to understand the stone tools, commonly regarded as “utilitarian” items. As a result, this class of grave goods has not received the same level of attention, leaving a significant gap in our understanding of Mesolithic mortuary behaviours. Our research challenges long-lasting perceptions of lithic tools as strictly utilitarian objects and draws on studies of one of the most substantial stone axe funerary collections from one of the largest Stone Age cemeteries in Europe–Zvejnieki, Latvia. Evidence suggests the selection of unused axes as grave offerings, while unusual wear traces on an axe found in a female grave (no 57) raises questions about its use in the burial rites. Using a multi-proxy approach, we compare life histories of axes placed in burials to those recovered from contemporary, nearby settlement contexts. Finally, a strong correlation between axes and women and children at Zvejnieki challenges gendered stereotypes of stone tools, historically regarded as possessions of the adult male members of Stone Age societies

Somatic mutations of CADM1 in aldosterone-producing adenomas and gap junction-dependent regulation of aldosterone production

Aldosterone-producing adenomas (APAs) are the commonest curable cause of hypertension. Most have gain-of-function somatic mutations of ion channels or transporters. Herein we report the discovery, replication and phenotype of mutations in the neuronal cell adhesion gene CADM1. Independent whole exome sequencing of 40 and 81 APAs found intramembranous p.Val380Asp or p.Gly379Asp variants in two patients whose hypertension and periodic primary aldosteronism were cured by adrenalectomy. Replication identified two more APAs with each variant (total, n = 6). The most upregulated gene (10- to 25-fold) in human adrenocortical H295R cells transduced with the mutations (compared to wildtype) was CYP11B2 (aldosterone synthase), and biological rhythms were the most differentially expressed process. CADM1 knockdown or mutation inhibited gap junction (GJ)-permeable dye transfer. GJ blockade by Gap27 increased CYP11B2 similarly to CADM1 mutation. Human adrenal zona glomerulosa (ZG) expression of GJA1 (the main GJ protein) was patchy, and annular GJs (sequelae of GJ communication) were less prominent in CYP11B2-positive micronodules than adjacent ZG. Somatic mutations of CADM1 cause reversible hypertension and reveal a role for GJ communication in suppressing physiological aldosterone production

Anticipatory governance for social-ecological resilience

Anticipation is increasingly central to urgent contemporary debates, from climate change to the global economic crisis. Anticipatory practices are coming to the forefront of political, organizational, and citizens’ society. Research into anticipation, however, has not kept pace with public demand for insights into anticipatory practices, their risks and uses. Where research exists, it is deeply fragmented. This paper seeks to identify how anticipation is defined and understood in the literature and to explore the role of anticipatory practice to address individual, social, and global challenges. We use a resilience lens to examine these questions. We illustrate how varying forms of anticipatory governance are enhanced by multi-scale regional networks and technologies and by the agency of individuals, drawing from an empirical case study on regional water governance of Malaren, Sweden. Finally, we discuss how an anticipatory approach can inform adaptive institutions, decision making, strategy formation, and societal resilience

8310 CACNA1D- and KCNJ5-Mutant Aldosterone-Producing Adenomas (APAs) Have Opposite 2-Year Clinical Outcomes from Adrenalectomy: Prospective Trial Findings Explained By Different Cells-Of-Origin

Disclosure: K. Laycock: None. C. Cabrera: None. E. Wozniak: None. E. Ng: None. X. Wu: None. E. Goodchild: None. E.A. Azizan: None. J. Boot: None. C. Mein: None. J. MacFarlane: None. M. Gurnell: None. W. Drake: None. M.J. Brown: None. In a recent trial, adrenalectomy achieved complete clinical success at 6 months in only 30% of 78 patients with unilateral primary aldosteronism (PA) (Nat Med 2023 29:190). We have now compared influence of common APA genotypes on success at 2 years, and sought different cells of origin to explain the result. Methods: Home BP, plasma aldosterone and renin were re-measured 2 years after surgery. APA genotype and transcriptome were analysed by RNAseq. Single-nucleus (Sn) RNAs from 17 fresh-frozen adrenals and 9 APAs (3 each with CACNA1D or KCNJ5-mutation), were quantified by 10x Chromium, and cells with mutations identified by targeted amplicon analysis. Unusual cells were confirmed by immunofluorescence (IF), and their origin sought in published datasets for other tissues. Results: 14/18 patients with KCNJ5-mutation, but 0/20 with CACNA1D, had BP <135/85 mmHg off treatment at 2 years (p=0.03, logistic regression on genotype, age, gender, ethnicity). Biochemical success rate was also lower in CACNA1D-mutant patients. The most upregulated gene in CACNAID vs KCNJ5-mutant APAs was the endothelial-progenitor CCM2L (33-fold, p=10-26), followed by neuronal and adrenomedullary genes e.g. PTPRZ1, SLC35F1, UNC79, by 13-17-fold, p=10-[1]0-[1]2). Transcripts of these genes, in Sn analyses of 85203 nuclei from 17 non-tumour samples, clustered in CYP11B2-expressing cells, along with genes characteristic of aldosterone-producing micronodules (APM). The APM cluster was downstream of medulla on pseudotime analysis. In Sn analysis of 9 APAs, CCM2L was highly differentiated in CYP11B2+ cells from 3 CACNA1D- vs 3 KCNJ5-mutants (p=10-[1]06) along with PTPRZ1 (p=10-232), SLC35F1 (p=0) and many other neuronal/medullary transcripts. UMAPs of APAs and adjacent adrenal both revealed a cluster of hybrid cells expressing CCM2L or other endothelial genes, plus CYP11B2 and other zona glomerulosa (ZG) or APM-selective transcripts. The proportion of hybrid cells was higher in CACNA1D-mutant APAs (236/803) and adjacent adrenal (37/946) than KCNJ5-mutant (101/954) and adjacent tissue (8/766). By contrast, CYP11B2+ cells from KCNJ5-mutant APAs were enriched for transcripts in the CYP11B2- ZG-cell cluster of adjacent adrenal. Amplicon analysis of APAs detected CACNA1D mutation in multiple cells of the endothelial cluster. IF confirmed cells expressing CYP11B2 and endothelial transcripts. Mining of GTEX, and Human Cell Atlas led to a cluster of CD34+RENBP+ endothelial progenitors in spleen as likely source of CCM2L+ cells (Gen Biol 2019 21:1). Conclusion: Somatic genotype influences long-term outcome after adrenalectomy for PA, reflecting origin of CACNA1D-, but not KCNJ5-, mutant APAs from APMs. Bilateral APMs have been inferred from their occurrence in most PA adrenals (Nat Rev Neph 2023 19:788). This is supported by our finding of CCM2L+CYP11B2+ hybrid cells, with a likely circulatory origin from spleen. Presentation: 6/3/202

8310 CACNA1D- and KCNJ5-Mutant Aldosterone-Producing Adenomas (APAs) Have Opposite 2-Year Clinical Outcomes from Adrenalectomy: Prospective Trial Findings Explained By Different Cells-Of-Origin

Disclosure: K. Laycock: None. C. Cabrera: None. E. Wozniak: None. E. Ng: None. X. Wu: None. E. Goodchild: None. E.A. Azizan: None. J. Boot: None. C. Mein: None. J. MacFarlane: None. M. Gurnell: None. W. Drake: None. M.J. Brown: None. In a recent trial, adrenalectomy achieved complete clinical success at 6 months in only 30% of 78 patients with unilateral primary aldosteronism (PA) (Nat Med 2023 29:190). We have now compared influence of common APA genotypes on success at 2 years, and sought different cells of origin to explain the result. Methods: Home BP, plasma aldosterone and renin were re-measured 2 years after surgery. APA genotype and transcriptome were analysed by RNAseq. Single-nucleus (Sn) RNAs from 17 fresh-frozen adrenals and 9 APAs (3 each with CACNA1D or KCNJ5-mutation), were quantified by 10x Chromium, and cells with mutations identified by targeted amplicon analysis. Unusual cells were confirmed by immunofluorescence (IF), and their origin sought in published datasets for other tissues. Results: 14/18 patients with KCNJ5-mutation, but 0/20 with CACNA1D, had BP <135/85 mmHg off treatment at 2 years (p=0.03, logistic regression on genotype, age, gender, ethnicity). Biochemical success rate was also lower in CACNA1D-mutant patients. The most upregulated gene in CACNAID vs KCNJ5-mutant APAs was the endothelial-progenitor CCM2L (33-fold, p=10-26), followed by neuronal and adrenomedullary genes e.g. PTPRZ1, SLC35F1, UNC79, by 13-17-fold, p=10-[1]0-[1]2). Transcripts of these genes, in Sn analyses of 85203 nuclei from 17 non-tumour samples, clustered in CYP11B2-expressing cells, along with genes characteristic of aldosterone-producing micronodules (APM). The APM cluster was downstream of medulla on pseudotime analysis. In Sn analysis of 9 APAs, CCM2L was highly differentiated in CYP11B2+ cells from 3 CACNA1D- vs 3 KCNJ5-mutants (p=10-[1]06) along with PTPRZ1 (p=10-232), SLC35F1 (p=0) and many other neuronal/medullary transcripts. UMAPs of APAs and adjacent adrenal both revealed a cluster of hybrid cells expressing CCM2L or other endothelial genes, plus CYP11B2 and other zona glomerulosa (ZG) or APM-selective transcripts. The proportion of hybrid cells was higher in CACNA1D-mutant APAs (236/803) and adjacent adrenal (37/946) than KCNJ5-mutant (101/954) and adjacent tissue (8/766). By contrast, CYP11B2+ cells from KCNJ5-mutant APAs were enriched for transcripts in the CYP11B2- ZG-cell cluster of adjacent adrenal. Amplicon analysis of APAs detected CACNA1D mutation in multiple cells of the endothelial cluster. IF confirmed cells expressing CYP11B2 and endothelial transcripts. Mining of GTEX, and Human Cell Atlas led to a cluster of CD34+RENBP+ endothelial progenitors in spleen as likely source of CCM2L+ cells (Gen Biol 2019 21:1). Conclusion: Somatic genotype influences long-term outcome after adrenalectomy for PA, reflecting origin of CACNA1D-, but not KCNJ5-, mutant APAs from APMs. Bilateral APMs have been inferred from their occurrence in most PA adrenals (Nat Rev Neph 2023 19:788). This is supported by our finding of CCM2L+CYP11B2+ hybrid cells, with a likely circulatory origin from spleen. Presentation: 6/3/202

8310 CACNA1D- and KCNJ5-Mutant Aldosterone-Producing Adenomas (APAs) Have Opposite 2-Year Clinical Outcomes from Adrenalectomy: Prospective Trial Findings Explained By Different Cells-Of-Origin

Disclosure: K. Laycock: None. C. Cabrera: None. E. Wozniak: None. E. Ng: None. X. Wu: None. E. Goodchild: None. E.A. Azizan: None. J. Boot: None. C. Mein: None. J. MacFarlane: None. M. Gurnell: None. W. Drake: None. M.J. Brown: None. In a recent trial, adrenalectomy achieved complete clinical success at 6 months in only 30% of 78 patients with unilateral primary aldosteronism (PA) (Nat Med 2023 29:190). We have now compared influence of common APA genotypes on success at 2 years, and sought different cells of origin to explain the result. Methods: Home BP, plasma aldosterone and renin were re-measured 2 years after surgery. APA genotype and transcriptome were analysed by RNAseq. Single-nucleus (Sn) RNAs from 17 fresh-frozen adrenals and 9 APAs (3 each with CACNA1D or KCNJ5-mutation), were quantified by 10x Chromium, and cells with mutations identified by targeted amplicon analysis. Unusual cells were confirmed by immunofluorescence (IF), and their origin sought in published datasets for other tissues. Results: 14/18 patients with KCNJ5-mutation, but 0/20 with CACNA1D, had BP <135/85 mmHg off treatment at 2 years (p=0.03, logistic regression on genotype, age, gender, ethnicity). Biochemical success rate was also lower in CACNA1D-mutant patients. The most upregulated gene in CACNAID vs KCNJ5-mutant APAs was the endothelial-progenitor CCM2L (33-fold, p=10-26), followed by neuronal and adrenomedullary genes e.g. PTPRZ1, SLC35F1, UNC79, by 13-17-fold, p=10-[1]0-[1]2). Transcripts of these genes, in Sn analyses of 85203 nuclei from 17 non-tumour samples, clustered in CYP11B2-expressing cells, along with genes characteristic of aldosterone-producing micronodules (APM). The APM cluster was downstream of medulla on pseudotime analysis. In Sn analysis of 9 APAs, CCM2L was highly differentiated in CYP11B2+ cells from 3 CACNA1D- vs 3 KCNJ5-mutants (p=10-[1]06) along with PTPRZ1 (p=10-232), SLC35F1 (p=0) and many other neuronal/medullary transcripts. UMAPs of APAs and adjacent adrenal both revealed a cluster of hybrid cells expressing CCM2L or other endothelial genes, plus CYP11B2 and other zona glomerulosa (ZG) or APM-selective transcripts. The proportion of hybrid cells was higher in CACNA1D-mutant APAs (236/803) and adjacent adrenal (37/946) than KCNJ5-mutant (101/954) and adjacent tissue (8/766). By contrast, CYP11B2+ cells from KCNJ5-mutant APAs were enriched for transcripts in the CYP11B2- ZG-cell cluster of adjacent adrenal. Amplicon analysis of APAs detected CACNA1D mutation in multiple cells of the endothelial cluster. IF confirmed cells expressing CYP11B2 and endothelial transcripts. Mining of GTEX, and Human Cell Atlas led to a cluster of CD34+RENBP+ endothelial progenitors in spleen as likely source of CCM2L+ cells (Gen Biol 2019 21:1). Conclusion: Somatic genotype influences long-term outcome after adrenalectomy for PA, reflecting origin of CACNA1D-, but not KCNJ5-, mutant APAs from APMs. Bilateral APMs have been inferred from their occurrence in most PA adrenals (Nat Rev Neph 2023 19:788). This is supported by our finding of CCM2L+CYP11B2+ hybrid cells, with a likely circulatory origin from spleen. Presentation: 6/3/202

[11C]metomidate PET-CT versus adrenal vein sampling for diagnosing surgically curable primary aldosteronism: a prospective, within-patient trial.

Primary aldosteronism (PA) due to a unilateral aldosterone-producing adenoma is a common cause of hypertension. This can be cured, or greatly improved, by adrenal surgery. However, the invasive nature of the standard pre-surgical investigation contributes to fewer than 1% of patients with PA being offered the chance of a cure. The primary objective of our prospective study of 143 patients with PA ( NCT02945904 ) was to compare the accuracy of a non-invasive test, [11C]metomidate positron emission tomography computed tomography (MTO) scanning, with adrenal vein sampling (AVS) in predicting the biochemical remission of PA and the resolution of hypertension after surgery. A total of 128 patients reached 6- to 9-month follow-up, with 78 (61%) treated surgically and 50 (39%) managed medically. Of the 78 patients receiving surgery, 77 achieved one or more PA surgical outcome criterion for success. The accuracies of MTO at predicting biochemical and clinical success following adrenalectomy were, respectively, 72.7 and 65.4%. For AVS, the accuracies were 63.6 and 61.5%. MTO was not significantly superior, but the differences of 9.1% (95% confidence interval = -6.5 to 24.1%) and 3.8% (95% confidence interval = -11.9 to 9.4) lay within the pre-specified -17% margin for non-inferiority (P = 0.00055 and P = 0.0077, respectively). Of 24 serious adverse events, none was considered related to either investigation and 22 were fully resolved. MTO enables non-invasive diagnosis of unilateral PA

Somatic mutations of CADM1 in aldosterone-producing adenomas and gap junction-dependent regulation of aldosterone production.

Funder: DH | NIHR | Efficacy and Mechanism Evaluation Programme (NIHR Efficacy and Mechanism Evaluation Programme); doi: https://doi.org/10.13039/501100001922Funder: British Heart Foundation (BHF); doi: https://doi.org/10.13039/501100000274Funder: RCUK | Medical Research Council (MRC); doi: https://doi.org/10.13039/501100000265Funder: Royal Society-Newton Advanced Research Fellow (NA170257/FF-2018-033Funder: Japan Society for the Promotion of Science KAKENHI grants 17K08680Funder: DH | NIHR | Health Services Research Programme (NIHR Health Services Research Programme); doi: https://doi.org/10.13039/501100001923Funder: NIHR Translational ResearchFunder: Biomedical Research Council, SingaporeFunder: the European Research Council under the European Union’s Horizon 2020 research and innovation program. Grant agreement No. 694913Funder: Japan Society for the Promotion of Science KAKENHI grants. Grant ref no. 18K07414Funder: European Research Council under the European Union’s Horizon 2020 research and innovation program. Grant agreement No. 633983Funder: Japan Society for the Promotion of Science KAKENHI grants. Grant ref. 18K07049 and 15K15113 Ministry of Education, Culture, Sports, Science and Technology-Supported Program for the Strategic Research Foundation at Private Universities 2015-19Aldosterone-producing adenomas (APAs) are the commonest curable cause of hypertension. Most have gain-of-function somatic mutations of ion channels or transporters. Herein we report the discovery, replication and phenotype of mutations in the neuronal cell adhesion gene CADM1. Independent whole exome sequencing of 40 and 81 APAs found intramembranous p.Val380Asp or p.Gly379Asp variants in two patients whose hypertension and periodic primary aldosteronism were cured by adrenalectomy. Replication identified two more APAs with each variant (total, n = 6). The most upregulated gene (10- to 25-fold) in human adrenocortical H295R cells transduced with the mutations (compared to wildtype) was CYP11B2 (aldosterone synthase), and biological rhythms were the most differentially expressed process. CADM1 knockdown or mutation inhibited gap junction (GJ)-permeable dye transfer. GJ blockade by Gap27 increased CYP11B2 similarly to CADM1 mutation. Human adrenal zona glomerulosa (ZG) expression of GJA1 (the main GJ protein) was patchy, and annular GJs (sequelae of GJ communication) were less prominent in CYP11B2-positive micronodules than adjacent ZG. Somatic mutations of CADM1 cause reversible hypertension and reveal a role for GJ communication in suppressing physiological aldosterone production