Multiplex imaging reveals spatially resolved DNA-damage response neighborhoods in TP53-mutated myelodysplastic neoplasms.

While increased DNA damage is a well-described feature of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), it is unclear whether all lineages and all regions of the marrow are homogeneously affected. In this study, we performed immunohistochemistry on formalin-fixed, paraffin-embedded whole-section bone marrow biopsies using a well-established antibody to detect pH2A.X (phosphorylated histone variant H2A.X) that recognizes DNA double-strand breaks. Focusing on TP53-mutated and complex karyotype MDS/AML, we find a greater pH2A.X+ DNA damage burden compared to TP53 wild-type neoplastic cases and non-neoplastic controls. To understand how double-strand breaks vary between lineages and spatially in TP53-mutated specimens, we applied a low-multiplex immunofluorescence staining and spatial analysis protocol to visualize pH2A.X+ cells with p53 protein staining and lineage markers. pH2A.X marked predominantly mid- to late-stage erythroids, whereas early erythroids and CD34+ blasts were relatively spared. In a prototypical example, these pH2A.X+ erythroids were organized locally as distinct colonies, and each colony displayed pH2A.X+ puncta at a synchronous level. This highly coordinated immunophenotypic expression was also seen for p53 protein staining and among presumed early myeloid colonies. Neighborhood clustering analysis showed distinct marrow regions differentially enriched in pH2A.X+/p53+ erythroid or myeloid colonies, indicating spatial heterogeneity of DNA-damage response and p53 protein expression. The lineage and architectural context within which DNA damage phenotype and oncogenic protein are expressed is relevant to current therapeutic developments that leverage macrophage phagocytosis to remove leukemic cells in part due to irreparable DNA damage. © 2024 The Pathological Society of Great Britain and Ireland.

Modality and terminology changes for behavioral health service delivery during the COVID-19 pandemic: a systematic review.

Introduction: The COVID-19 pandemic prompted healthcare professionals to implement service delivery adaptations to remain in compliance with safety regulations. Though many adaptations in service delivery were reported throughout the literature, a wide variety of terminology and definitions were used. Methods: To address this, we conducted a PRISMA review to identify service delivery adaptations across behavioral healthcare services in the United States from March 2020 to May 2022 and to identify variations in terminology used to describe these adaptations. We identified 445 initial articles for our review across eight databases using predetermined keywords. Using a two-round screening process, authors used a team approach to identify the most appropriate articles for this review. Results: Our results suggested that a total of 14 different terms were used to describe service modality changes, with the most frequent term being telehealth (63%). Each term found in our review and the frequency of use across identified articles is described in detail. Discussion: Implications of this review such as understanding modality changes during the COVID-19 pandemic and beyond are discussed. Our findings illustrate the importance of standardizing terminology to enhance communication and understanding among professionals.

"Landmark" Articles in Female Pelvic Medicine and Reconstructive Surgery: Do We Read What We Cite?

OBJECTIVE: The aim of the study was to describe the provision of a "landmark" article reading list to trainees by their Female Pelvic Medicine & Reconstructive Surgery (FPMRS) fellowship programs. METHODS: In this cross-sectional study, a web-based survey was sent to program coordinators of accredited FPMRS programs. Available reading lists were collected, and a master list compiled. Nonparametric tests were used to investigate the association between the number of faculty members and presence of a reading list. RESULTS: We received a response from 46 (92%) of the 50 Accreditation Council for Graduate Medical Education programs contacted. Half the programs (23/46, 50%) reported that they do not offer a recommended reading list, 5 programs were unsure and over more than 60% with a list (12/18, 67%) shared their list with the investigators. Programs had between 2 and 14 core faculty members with a median (interquartile range) of 4 (3-6). Programs with a reading list had significantly fewer faculty members (3.5, 3-5 vs 5, 3-6, P = 0.03). There were 323 unique articles across the 12 lists, with a median of 25 articles per list. Thirty-four articles were on 4 or more lists; 9 of these (9/34, 26%) have previously been identified as citation classics. CONCLUSIONS: At least 50% of the Accreditation Council for Graduate Medical Education accredited programs do not provide their fellows with a recommended reading list, and fewer core faculty members are associated with the presence of such a list. Although some commonalities are found between reading lists, a universal reading list of "landmark" articles could improve the consistency of FPMRS fellowship education.

Mapping Critical Residues in ATG11's Coiled-Coil 2 Domain that Block Multiple Interactions and Disrupt Selective Autophagy.

Selective autophagy is a conserved subcellular process that maintains the health of eukaryotic cells by targeting damaged or toxic cytoplasmic components to the vacuole/lysosome for degradation. A key player in the initiation of selective autophagy in S. Cerevisiae (baker's yeast) is a large adapter protein called Atg11. Atg11 has multiple predicted coiled-coil domains and intrinsically disordered regions, is known to dimerize, and binds and organizes other essential components of the autophagosome formation machinery, including Atg1 and Atg9. We performed systematic directed mutagenesis on the coiled-coil 2 domain of Atg11 in order to map which residues were required for its structure and function. Using yeast-2-hybrid and coimmunoprecipitation, we found only three residues to be critical: I562, Y565, and I569. Mutation of any of these, but especially Y565, could interfere with Atg11 dimerization and block its interaction with Atg1 and Atg9, thereby inactivating selective autophagy.

High variance in community structure and ecosystem carbon stocks of Fijian mangroves driven by differences in geomorphology and climate.

Mangrove ecosystems are particularly important for small island developing states of the Pacific, such as Fiji, which are at the forefront of the impacts of climate change. This is because of the ability of mangroves to mitigate storm surges and floods as well as their high carbon sequestration and storage capacity. However, there are few detailed studies on the spatial variation in mangrove structure and carbon stocks in Fiji, and this information is essential to support decision making by government and communities, enabling the development of effective mitigation and adaptation responses. We assessed mangrove forest structure in contrasting regions around Fiji's largest island, Viti Levu, within sites managed by indigenous (iTaukei) Fijians. Mangroves of the Ba, Nadroga-Navosa, and Rewa and Tailevu regions showed high variance in both structural complexity and ecosystem carbon stocks. Levels of variation were similar to that observed globally due to variable geomorphological and biophysical settings related to orographic rainfall, freshwater influx, tidal amplitude and cyclonic disturbances. High biomass, structurally complex forests occur on the wetter south-east coast (e.g. the Rewa Delta), while structurally uniform scrub mangroves dominate large areas of mangroves along the north-west (e.g. the Ba Delta) and west coast (e.g. the Tuva Delta). Mangroves of the Ba region displayed considerable damage from tropical cyclones, particularly in taller vegetation. All mangrove sites assessed were important reservoirs of carbon, with results when scaled to the spatial extent of mangroves in Fiji revealing that ecosystem carbon storage is disproportionate to area and equates to 73.3% of the carbon held within terrestrial rainforests, despite occupying just 7.3% of the total area. This underscores the importance of mangroves as valuable carbon sinks in Fiji and the need to develop incentives for improved conservation and restoration.

Total Laparoscopic Hysterectomy of a 7400 g Uterus.

OBJECTIVE: To demonstrate improved techniques and safety measures for total laparoscopic hysterectomy for a severely enlarged uterus. DESIGN: Technical video of the surgery, which demonstrates various techniques for improved visualization, hemostasis, and manipulation for total laparoscopic hysterectomy for a 7400 g uterus. SETTING: Academic tertiary care hospital, University of Louisville Hospital, Louisville, Kentucky. INTERVENTIONS: A 44-year-old woman, gravida 0, presented with a severely enlarged myoma uterus, class III obesity with a body mass index of 40.4 kg/m2, and hypertension seeking minimally invasive hysterectomy. Magnetic resonance imaging demonstrated a uterus measuring 26 × 26 × 17 cm with multiple myomas. The patient was counseled in detail regarding minimally invasive surgery. She underwent total laparoscopic hysterectomy, bilateral salpingectomy, right oophorectomy, and cystoscopy. The total operative time was 4 hours 12 minutes, and the estimated blood loss was 700 mL. Pre- and postoperative hemoglobin was 13.3 g/dL and 11.3 g/dL, respectively. A 4-cm minilaparotomy incision was created as an extension of the umbilical port, and the specimen was extracted by scalpel morcellation using the extracorporeal C-incision tissue extraction technique. The patient was discharged home on postoperative day 1 and recovered without any complications. CONCLUSION: There is an increasing trend in performing laparoscopic hysterectomy for large uteri, which has a lower incidence of overall complications than laparotomy [1,2]. In addition, a cost analysis has demonstrated the superiority of laparoscopic hysterectomy for myomatous uterus accounting for the rare incidence of leiomyosarcoma [3]. Our video demonstrates improved hemostasis and visualization techniques through the use of high-cephalad camera and assistant ports, generous traditional bipolar desiccation, blunt retraction with suction irrigator in the midline umbilical port, and surgical bed rotation. We also describe in detail the scalpel morcellation technique. To date, we believe this is the largest uterine size removed laparoscopically that has been reported in the literature. In the hands of an experienced surgeon and with the demonstrated techniques, a laparoscopic approach to hysterectomy of a very enlarged uterus is safe and feasible.

Citation classics: the 100 most cited articles in Urogynecology.

INTRODUCTION AND HYPOTHESIS: The objective of this study was to utilize objective citation analyses to describe the 100 most cited articles in the field of Female Pelvic Medicine & Reconstructive Surgery (FPMRS) and to review their characteristics. METHODS: We searched the Thomas Reuters Web of Science database for the most cited articles within all journals classified as Obstetrics and Gynecology (Ob/Gyn), Urology (Uro), Colorectal Surgery (CRS), and vital General Medicine (GM) journals (New England Journal of Medicine; Journal of the American Medical Association; Lancet; The BMJ). We reviewed search results for FPMRS content and compiled a list of the top 100 most cited articles relevant to FPMRS. RESULTS: We screened 34,934 articles yielded by the initial search and compiled the 100 most cited articles relevant to FPMRS. Of these 100 titles, 40% (42 out of 105) were published in Ob/Gyn, 46.7% (49 out of 105) in Uro, 1.9% (2 out of 105) in CRS, and 11.4% (12 out of 105) in GM journals. The most cited FPMRS article was "The standardisation of terminology of lower urinary tract function: report from the Standardisation Sub-committee of the International Continence Society" (3,810 total and 242 citations per year). Over half the articles were observational in nature and the majority (58%) of them were related to urinary incontinence in women. CONCLUSIONS: Highly cited FPMRS articles come from a variety of journals, and nearly 50% of the 100 most cited FPMRS articles are from the urology literature. The most cited articles were largely observational rather than interventional studies and mostly related to female urinary incontinence.

Steric interference from intrinsically disordered regions controls dynamin-related protein 1 self-assembly during mitochondrial fission.

The self-assembling, mechanoenzymatic dynamin superfamily GTPase, dynamin-related protein 1 (Drp1), catalyzes mitochondrial and peroxisomal fission. Distinct intrinsically disordered regions (IDRs) in Drp1 substitute for the canonical pleckstrin homology (PH) domain and proline-rich domain (PRD) of prototypical dynamin, which cooperatively regulate endocytic vesicle scission. Whether the Drp1 IDRs function analogously to the corresponding dynamin domains however remains unknown. We show that an IDR unique to the Drp1 GTPase (G) domain, the 'extended 80-loop', albeit dissimilar in location, structure, and mechanism, functions akin to the dynamin PRD by enabling stable Drp1 mitochondrial recruitment and by suppressing Drp1 cooperative GTPase activity in the absence of specific partner-protein interactions. Correspondingly, we find that another IDR, the Drp1 variable domain (VD), in conjunction with the conserved stalk L1N loop, functions akin to the dynamin PH domain; first, in an 'auto-inhibitory' capacity that restricts Drp1 activity through a long-range steric inhibition of helical inter-rung G-domain dimerization, and second, as a 'fulcrum' for Drp1 self-assembly in the proper helical register. We show that the Drp1 VD is necessary and sufficient for specific Drp1-phospholipid interactions. We further demonstrate that the membrane-dependent VD conformational rearrangement essential for the alleviation of Drp1 auto-inhibition is contingent upon the basal GTP hydrolysis-dependent generation of Drp1 dimers from oligomers in solution. IDRs thus conformationally couple the enzymatic and membrane activities of Drp1 toward membrane fission.