bluuebunny/crossref_metadata_2025 huggingface dataset:
Advanced systemic mastocytosis—Revised classification, new drugs and how we treat
Authors: Pardanani Animesh , Reichard Kaaren , Tefferi Ayalew
Published: December 2023

Mastocytosis constitutes the neoplastic proliferation of mast cells and is broadly classified into systemic mastocytosis (SM), cutaneous mastocytosis and mast cell sarcoma. SM is further partitioned into advanced (AdvSM) and non‐advanced (SM‐non‐Adv) subcategories. AdvSM includes aggressive SM (ASM), SM with an associated haematological neoplasm (SM‐AHN) and mast cell leukaemia (MCL). In 2022, two separate expert committees representing the 5th edition of the World Health Organization (WHO5) and the International Consensus (ICC) classification systems submitted revised classification criteria for SM, highlighted by the ICC‐proposed incorporation of mast cell cytomorphology in the diagnostic criteria for MCL and myeloid‐lineage restriction for the AHN component in SM‐AHN. Recent developments in SM also include the introduction of KIT‐targeting tyrosine kinase inhibitors (KITi), including midostaurin and avapritinib, both drugs have shown potent activity in reducing mast cell and mutant KIT burden and alleviating mast cell‐associated organopathy and mediator symptoms; however, their overall impact on survival or superiority over pre‐KITi era treatment options (e.g. cladribine) has not been studied in a controlled setting. In the current review, we provide a summary of recent changes in disease classification and an analysis of recent clinical trials and their impact on our current treatment approach in AdvSM.
“What a tale we have been in”: Emplotment and the Exemplar Characters in The Lord of the Rings and the Harry Potter Series
Authors: Milbank Alison
Published: December 2023

Linda Zagzebski's theory of moral exemplarity emphasizes the importance of admiration in developing ethical behavior. This essay argues that admiration involves wonder and distance and is best evoked by mixed or flawed characters; it demonstrates this through discussion of the characters in J. R. R. Tolkien's The Lord of the Rings and J. K. Rowling's Harry Potter series. Using Paul Ricoeur's taxonomy of prefiguration, configuration, and refiguration in narrative work, it discerns a self‐reflexivity in the protagonists of these fantasy novels, which is echoed by that of the readers, who are brought to realize their own emplotment in larger narratives. Features in Tolkien and Rowling that aid this exploratory reading include the length and depth of the novels, the decentering of the reader's own reality, and their open endings, which offer an invitational role to further interpretation. Virtue is viewed more teleologically than in Zagzebski, for moral realism is woven into the metaphysics of these novels, which allows mimesis of flawed characters to be ethically productive.
Anoxia begets anoxia: A positive feedback to the deoxygenation of temperate lakes
Authors: Lewis Abigail S. L. , Lau Maximilian P. , Jane Stephen F. , Rose Kevin C. , Be'eri‐Shlevin Yaron , Burnet Sarah H. , Clayer François , Feuchtmayr Heidrun , Grossart Hans‐Peter , Howard Dexter W. , Mariash Heather , Delgado Martin Jordi , North Rebecca L. , Oleksy Isabella , Pilla Rachel M. , Smagula Amy P. , Sommaruga Ruben , Steiner Sara E. , Verburg Piet , Wain Danielle , Weyhenmeyer Gesa A. , Carey Cayelan C.
Published: December 2023

Declining oxygen concentrations in the deep waters of lakes worldwide pose a pressing environmental and societal challenge. Existing theory suggests that low deep‐water dissolved oxygen (DO) concentrations could trigger a positive feedback through which anoxia (i.e., very low DO) during a given summer begets increasingly severe occurrences of anoxia in following summers. Specifically, anoxic conditions can promote nutrient release from sediments, thereby stimulating phytoplankton growth, and subsequent phytoplankton decomposition can fuel heterotrophic respiration, resulting in increased spatial extent and duration of anoxia. However, while the individual relationships in this feedback are well established, to our knowledge, there has not been a systematic analysis within or across lakes that simultaneously demonstrates all of the mechanisms necessary to produce a positive feedback that reinforces anoxia. Here, we compiled data from 656 widespread temperate lakes and reservoirs to analyze the proposed anoxia begets anoxia feedback. Lakes in the dataset span a broad range of surface area (1–126,909 ha), maximum depth (6–370 m), and morphometry, with a median time‐series duration of 30 years at each lake. Using linear mixed models, we found support for each of the positive feedback relationships between anoxia, phosphorus concentrations, chlorophyll a concentrations, and oxygen demand across the 656‐lake dataset. Likewise, we found further support for these relationships by analyzing time‐series data from individual lakes. Our results indicate that the strength of these feedback relationships may vary with lake‐specific characteristics: For example, we found that surface phosphorus concentrations were more positively associated with chlorophyll a in high‐phosphorus lakes, and oxygen demand had a stronger influence on the extent of anoxia in deep lakes. Taken together, these results support the existence of a positive feedback that could magnify the effects of climate change and other anthropogenic pressures driving the development of anoxia in lakes around the world.
Relationship between a gum‐chewing routine and oral, physical, and cognitive functions of community‐dwelling older adults: A Kashiwa cohort study
Authors: Kawamura Jun , Tanaka Tomoki , Kanno Susumu , Osawa Kenji , Okabayashi Kazuto , Hirano Hirohiko , Shirobe Maki , Nagatani Miyuki , Son Bo‐Kyung , Lyu Weida , Iijima Katsuya
Published: December 2023

Aim

As associations between oral function and general health have been reported in community‐dwelling older adults, easily implementable preventive measures are urgently required. We focused on the health benefits of gum chewing, as no studies have been carried out on the impact of gum‐chewing routines on the health of older adults. This cross‐sectional study aimed to determine whether the gum‐chewing routine is associated with oral, physical and cognitive functions in community‐dwelling older adults.

Methods

This study included 1617 community‐dwelling older participants in a health survey carried out in 2021. The gum‐chewing routine and weekly chewing time were assessed using a self‐administered questionnaire. The outcome measures, including actual measurements of oral function, physical function, cognitive function, dietary intake and lifestyle, were evaluated using self‐administered questionnaires or health surveys.

Results

We analyzed 1474 (mean age 76.1 ± 5.8 years, 45% women) participants for whom all data were not missing, and 14% of them had a gum‐chewing routine for more than 30 min weekly. Oral functions were significantly higher in older adults with a gum‐chewing routine, and there were substantially fewer participants with oral frailty (adjusted odds ratio 0.581, 95% confidence interval 0.340–0.993). Additionally, cognitive and physical functions, including grip strength, were significantly higher in the gum‐chewing routine group.

Conclusions

Community‐dwelling older adults with a gum‐chewing routine have higher oral, physical and cognitive functions. These findings indicate that a gum‐chewing routine might contribute to maintaining oral function and preventing frailty. Geriatr Gerontol Int 2024; 24: 68–74.
Circular RNA circVAPA modulates macrophage pyroptosis in sepsis‐induced acute lung injury through targeting miR‐212‐3p/Sirt1/Nrf2/NLRP3 axis
Authors: Huang Yanjing , Lin Jinquan , Wu Zhiwei , Li Yiming
Published: December 2023

Sepsis‐induced acute lung injury (ALI) is an inflammatory condition involving the pyroptosis of macrophages. This study investigated the role of circular RNA hsa_circ_0006990 (circVAPA) in regulating macrophage pyroptosis in ALI and the underlying mechanisms. The expression pattern of circVAPA was examined in the mouse model of ALI and in the LPS‐treated RAW264.7 macrophage cell line. Lung tissue damage was evaluated by haematoxylin and eosin staining, immunohistochemistry and a myeloperoxidase activity assay. The molecular mechanisms were investigated by luciferase reporter assay, western blot, RT‐qPCR and ELISA. circVAPA was down‐regulated in the lung tissues of ALI mice and LPS‐induced RAW264.7 cells. circVAPA over‐expression alleviated lung tissue injury and dampened LPS‐induced pyroptosis and Th17‐associated inflammatory responses. miR‐212‐3p was identified as a target of circVAPA, and miR‐212‐3p negatively regulated the expression of Sirt1. Sirt1 knockdown largely abolished the effect of circVAPA over‐expression on pyroptosis. CircVAPA/miR‐212‐3p/Sirt1 axis also regulates Nrf2 and NLRP3 expression upon LPS challenge. By targeting miR‐212‐3p, circVAPA over‐expression negatively regulates the expression of Sirt1 and pyroptosis‐related factors (Nrf2 and NLRP3), which alleviates the inflammatory damages in sepsis‐induced ALI.
Three‐dimensional representation of the Vita Toothguide 3D‐Master: An in vivo clinical study
Authors: Gómez‐Polo Cristina , Fraile Javier Flores , López Norberto Quispe , Muñoz Maria Portillo , Lobato Maria , Montero Javier
Published: December 2023

Statement of Problem

The Vita Toothguide 3D‐Master (Vita Zahnfabrik) is considered the dental shade guide in which the three dimensions of color – lightness, hue, and chroma – are most well‐ordered in the CIELAB color space. No research has yet explored how well the Vita Toothguide 3D‐Master is ordered in the 3D color space by recording color coordinates in vivo.

Purpose

To evaluate the spatial color distribution of the Vita Toothguide 3D‐Master's lightness, chroma, and hue groups and its 26 physical shade tabs.

Materials and Methods

The dental color (L*, C*, h°, a*, and b* color coordinates) of a healthy maxillary central incisor was recorded for 1361 participants (aged between 18 and 89 years) using a Vita Easyshade Compact spectrophotometer (Vita Zahnfabrik). The R 2.7.2. statistics program was used to create the visual representations.

Results

The five lightness levels are those that are best distributed in the color space, in relation to the L* coordinates, followed by the chroma group and, finally, the hue group. The 5M1, 5M2, and 5M3 physical shade tabs are situated at a greater distance apart from the other tabs in the color space.

Conclusions

The Vita Toothguide 3D‐Master's 26 physical shade tabs are satisfactorily distributed in three‐dimensional space, although strict mathematical criteria are not followed. The natural dental shades that fall lower on the lightness scale are the most poorly represented by the physical shade tabs.

Clinical Implications

Darker teeth are poorly represented by the Vita Toothguide 3D‐Master's physical shade tabs. The spatial distribution of dental shade guides needs to be improved to ensure they provide homogeneous coverage of the entire chromatic spectrum corresponding to natural teeth. This would help reduce the errors inherent to the subjective visual color selection process.
The association between orthodontic treatment and temporomandibular disorders diagnosis and disease characteristics
Authors: Shalish Miriam , Leibovich Avi , Zakuto Avraham , Slutzky Hulio , Chaushu Stella , Almoznino Galit
Published: December 2023

Background

The association between orthodontic treatment and temporomandibular disorders (TMD) has been contentious in the literature.

Objectives

To analyse the associations between orthodontic treatment and TMD diagnosis and diseases characteristics.

Methods

This case–control study included 291 individuals, 192 TMDs and 99 controls. All patients underwent assessment based on a questionnaire and a clinical examination according to Axis I of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Differences in orthodontic treatment between TMDs and controls, as well as across different TMD diagnoses within the TMD group were analysed. Patients who underwent orthodontic treatment were compared to those who did not, regarding their trauma history, bruxism, aggressive teeth brushing, level of oral hygiene, pain scores, muscle tenderness scores and subjective sleep quality.

Results

Of the 291 participants, 119 (40.9%) underwent orthodontic treatment and 172 (59.1%) had no orthodontics experience. Orthodontic treatment included: mandibular orthodontic treatment (102 subjects) and maxillary orthodontic treatment (113 subjects) of those 47 used a headgear.

Following multivariate analysis among TMDs, orthodontic treatment was associated with a good level of oral hygiene versus poor (Odds ratio [OR]: 5.17 [1.04–25.59]), lower number of tender muscles [OR = 0.84 (0.74–0.96)] and lower (better) Pittsburgh Sleep Quality Index (PSQI) scores (OR = 0.86 [0.76–0.97]). None of the studied parameters maintained a statistically significant association with orthodontic treatment in the multivariate analysis among the entire study population.

Conclusions

Utilising a holistic approach, this study supports the main hypothesis that orthodontic treatment is not associated with TMD diagnosis and disease characteristics.
Dietary effects of microalga Tetraselmis suecica on growth, antioxidant‐immune activity, inflammation cytokines, and resistance of Nile tilapia fingerlings to Aeromonas sobria infection
Authors: Abdel‐Tawwab Mohsen , Khalil Riad H. , Abdel‐Razek Nashwa , Younis Nehal A. , Shady Sherien H. H. , Monier Mohamed N. , Abdel‐Latif Hany M. R.
Published: December 2023

The dietary effects of the green microalga Tetraselmis suecica (TS) on the growth, digestive enzymes, immune and antioxidant responses, genes expression, and disease resistance of Nile tilapia (Oreochromis niloticus) fingerlings were investigated. This microalga was mixed with the diet' ingredients at doses of 0.0 (the control), 5, 10, 15, and 20 g/kg diet and then fed to fish daily for 84 days. After the feeding trial, fish were experimentally challenged with Aeromonas sobria, infection and fish mortalities were recorded for another 10 days. Dietary TS significantly (p < 0.05) enhanced growth, digestive enzymes activities, and blood proteins, particularly at the level of 15 g/kg diet. Feeding the fish on 15 TS/kg feed exhibited highest mRNA expressions of GH and IGF‐1 genes as well as SOD, CAT, and GPx genes compared to other TS groups. Moreover, highest levels of hepatic antioxidant and immune indices were found in the treatment of 15 g TS/kg feed. Significant downregulation of IL‐1β and IL‐8 genes expression and significant upregulation of IL‐10 gene expression were observed in TS‐fed fish, principally in fish groups fed on 15−20 g TS/kg feed. Conversely, hepatic malondialdehyde levels, blood glucose, and the activities of transaminases (ALT and AST) were significantly (p < 0.05) decreased in fish fed with 15−20 g TS/kg diet. Serum bactericidal activity against A. sobria was significantly higher in TS‐fed fish groups, and its highest levels were found in treatments of 15−20 g/kg diet. Of interest, the survival rates of fish groups fed diets with 10−20 g TS/kg feed were higher after the challenge with A. sobria infection than the control group. Accordingly, we can conclude that supplementing fish diets with a 15 g TS/kg diet enhanced the growth, antioxidant and immune activities, and resistance of Nile tilapia fingerlings to possible A. sobria infection.
The delayed impact of informed versus blind interviewing on eyewitness memory
Authors: Rivard Jillian , Carlson Victoria , LaBat Devon E. , Compo Nadja Schreiber
Published: December 2023

Background

Previous research has highlighted the potentially detrimental effects of pre‐interview preparation on witness memory within an interview context (Rivard et al., 2016). The present study examined the effect of an interviewer's pre‐interview knowledge on eyewitness memory beyond the initial interview.

Method

Student witnesses were interviewed one week after viewing a mock crime event by a student interviewer who was either correctly informed, incorrectly informed, or uninformed (blind) to case details and who was either told to avoid suggestions or was not given cautionary instructions.

Results

Analyses of the witnesses' recall quantity and quality one week after the interview revealed that witnesses of blind interviewers recalled more details than witnesses of incorrectly informed interviewers. Witnesses of blind interviewers were also more accurate than witnesses of incorrectly informed interviewers, but only when interviewers were warned not to ask suggestive questions.

Implications

Findings suggest that interviewer training and pre‐interview knowledge may play an important role in witness recall.
FLZ13 interacts with FLC and ABI5 to negatively regulate flowering time in Arabidopsis
Authors: Li Xibao , Lai Minyi , Li Kailin , Yang Lianming , Liao Jun , Gao Yingmiao , Wang Ying , Gao Caiji , Shen Wenjin , Luo Ming , Yang Chao
Published: December 2023

The transition from vegetative to reproductive growth, known as flowering, is a critical developmental process in flowering plants to ensure reproductive success. This process is strictly controlled by various internal and external cues; however, the underlying molecular regulatory mechanisms need to be further characterized.

Here, we report a plant‐specific protein, FCS‐LIKE ZINC FINGER PROTEIN 13 (FLZ13), which functions as a hitherto unknown negative modulator of flowering time in Arabidopsis thaliana.

Biochemical analysis showed that FLZ13 directly interacts with FLOWERING LOCUS C (FLC), a major flowering repressor, and that FLZ13 largely depends on FLC to repress the transcription of two core flowering integrators: FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1. In addition, FLZ13 works together with ABSCISIC ACID INSENSITIVE 5 to activate FLC expression to delay flowering.

Taken together, our findings suggest that FLZ13 is an important component of the gene regulatory network for flowering time control in plants.
Photocatalytic fixation and oxygenation of NAD+/NADP+ and sulfides using solar light: Exploring mechanistic investigations and their impact on synthetic applications
Authors: Shukla Ravindra K. , Yadav Rajesh K. , Gole Vittal L. , Singh Satyam , Gupta Navneet Kumar , Baeg Jin‐Ook
Published: December 2023

Sulfur‐doped Eosin‐B (SDE‐B) photocatalysts were synthesized for the first time utilizing sublimed sulfur (S8) as a dopant in an in situ thermal copolymerization technique. Sulfur doping not only increased Eosin‐B (E‐B) absorption range for solar radiation but also improved fixation and oxygenation capabilities. The doped sulfur bridges the S‐S bond by substituting for the edge bromine of the E‐B bond. The improved photocatalytic activity of SDE‐B in the fixation and oxygenation of NAD+/NADP+ and sulfides using solar light is attributed to the photo‐induced hole of SDE‐B's high fixation and oxygenation capacity, as well as an efficient suppression of electron and hole recombination. The powerful light‐harvesting bridge system created using SDE‐B as a photocatalyst works extremely well, resulting in high NADH/NADPH regeneration (79.58/76.36%) and good sulfoxide yields (98.9%) under solar light. This study focuses on the creation and implementation of a sulfur‐doped photocatalyst for direct fine chemical regeneration and organic transformation.
Approximating Botswana's financial cycle: Expanding the macroprudential toolkit
Authors: Setshegetso Leonard Nnete , Mado Mogakolodi
Published: December 2023

This study estimates the financial cycle for Botswana using the Christiano–Fitzgerald band‐pass filter, unobserved components approach and a Markov switching dynamic factor model. Using real credit and equity prices, together with several macroeconomic variables from 2001Q1 to 2021Q4, we find that the domestic financial cycle generally captures movements along the business cycle, with peaks reflecting the cumulative build‐up of risks during the boom that ultimately burst, coinciding with periods of financial distress. These findings shed light on the implications of financial fluctuations on domestic financial stability, hence, inform effective calibration of macroprudential tools, while also providing traction for implementation of the recently approved macroprudential policy framework for Botswana.
Access to Higher Education and Adolescent Fertility in Chile
Authors: Salinas Viviana , Jorquera‐Samter Valentina , Wiegand‐Cruz Pilar
Published: December 2023

This study investigates gender differences in the association between adolescent fertility and the likelihood of initiating higher education among young Chilean men and women. We adopt an entropy balancing strategy to estimate the association between adolescent fertility and the likelihood of starting higher education while accounting for potential selection into early childbearing due to socioeconomic status and prior academic achievement. We use data from official national registers that cover a cohort of Chilean students who attended publicly funded schools and who successfully completed secondary schooling between 2011 and 2022. Our results indicate that adolescent mothers are 15 percentage points less likely to initiate higher education than their peers who did not give birth during adolescence. In comparison, teenage fathers are 20 percentage points less likely to do so than their childless counterparts. Our findings stand in contrast to previously identified disadvantage patterns for secondary school completion, whereby adolescent fertility more significantly hinders schooling completion for women relative to men. We contend that this reversal may be related to traditional gender‐role expectations in Chile, which encourage young fathers to act as providers and, therefore, may be prevented from continuing on their education path into tertiary studies.
From healthcare professional to degree apprentice
Authors: Picts Alexandra , Russell Anne‐Marie
Published: December 2023

Background

In the UK, new degree apprenticeship opportunities are enabling non‐medical practitioners to develop advanced roles. Frameworks to structure and standardise this development are also becoming more common. Knowing that historically healthcare professionals moving into advanced roles have experienced a transition period, we undertook a qualitative study to explore how this role transition – from healthcare professional to Advanced Clinical Practitioner (ACP) – was experienced in a degree apprentice programme.

Methods

First year ACP degree apprentices were purposively selected from a cohort of 28 enrolled on a 3‐year Masters programme at a UK University in 2021/22. Consenting participants took part in in‐depth qualitative online semi‐structured interviews, which were recorded, transcribed and analysed thematically.

Findings

The five participants were in their first year of ACP training and represented core professional groups and primary, secondary and tertiary healthcare sectors. Five themes were identified: 1. what ACP apprentices bring; 2. reflections on how they see themselves; 3. how others see them; 4. effects of employing organisation's support; and 5. experience of Masters level learning.

Conclusion

ACP apprentices experienced a strong and often difficult transition period at the beginning of their training. The themes identified as influencing this could be used by higher education providers and clinical workplaces to better understand this period in training. Specifically, having a clear transition point to a defined role; a reduced workload during the transition period; and improved information sharing to better prepare workplaces for trainees, could all improve the experience.
Immunity-linked genes are stimulated by a membrane stress pathway linked to Golgi function and the ARF-1 GTPase
Authors: Fanelli Matthew J. , Welsh Christofer M. , Lui Dominique S. , Smulan Lorissa J. , Walker Amy K.
Published: December 2023

Infection response and other immunity-linked genes (ILGs) were first named in
Caenorhabditis elegans
–based expression after pathogen challenge, but many are also up-regulated when lipid metabolism is perturbed. Why pathogen attack and metabolic changes both increase ILGs is unclear. We find that ILGs are activated when phosphatidylcholine (PC) levels change in membranes of secretory organelles in
C. elegans
. RNAi targeting of the ADP-ribosylation factor
arf-1
, which disrupts the Golgi and secretory function, also activates ILGs. Low PC limits ARF-1 function, suggesting a mechanism for ILG activation via lipid metabolism, as part of a membrane stress response acting outside the ER. RNAi of selected ILGs uncovered defects in the secretion of two GFP reporters and the accumulation of a pathogen-responsive complement C1r/C1s, Uegf, Bmp1 (CUB) domain fusion protein. Our data argue that up-regulation of some ILGs is a coordinated response to changes in trafficking and may act to counteract stress on secretory function.
RAD54L2-mediated DNA damage avoidance pathway specifically preserves genome integrity in response to topoisomerase 2 poisons
Authors: Zhang Huimin , Xiong Yun , Sun Yilun , Park Jeong-Min , Su Dan , Feng Xu , Keast Sarah , Tang Mengfan , Huang Min , Wang Chao , Srivastava Mrinal , Yang Chang , Zhu Dandan , Chen Zhen , Li Siting , Yin Ling , Pommier Yves , Chen Junjie
Published: December 2023

Type II topoisomerases (TOP2) form transient TOP2 cleavage complexes (TOP2ccs) during their catalytic cycle to relieve topological stress. TOP2ccs are covalently linked TOP2-DNA intermediates that are reversible but can be trapped by TOP2 poisons. Trapped TOP2ccs block transactions on DNA and generate genotoxic stress, which are the mechanisms of action of TOP2 poisons. How cells avoid TOP2cc accumulation remains largely unknown. In this study, we uncovered RAD54 like 2 (RAD54L2) as a key factor that mediates a TOP2-specific DNA damage avoidance pathway. RAD54L2 deficiency conferred unique sensitivity to treatment with TOP2 poisons. RAD54L2 interacted with TOP2A/TOP2B and ZATT/ZNF451 and promoted the turnover of TOP2 from DNA with or without TOP2 poisons. Additionally, inhibition of proteasome activity enhanced the chromatin binding of RAD54L2, which in turn led to the removal of TOP2 from chromatin. In conclusion, we propose that RAD54L2-mediated TOP2 turnover is critically important for the avoidance of potential TOP2-linked DNA damage under physiological conditions and in response to TOP2 poisons.
Distinct beta-arrestin coupling and intracellular trafficking of metabotropic glutamate receptor homo- and heterodimers
Authors: Lee Joon , Gonzalez-Hernandez Alberto J. , Kristt Melanie , Abreu Nohely , Roßmann Kilian , Arefin Anisul , Marx Dagan C. , Broichhagen Johannes , Levitz Joshua
Published: December 2023

The metabotropic glutamate receptors (mGluRs) are family C, dimeric G protein–coupled receptors (GPCRs), which play critical roles in synaptic transmission. Despite an increasing appreciation of the molecular diversity of this family, how distinct mGluR subtypes are regulated remains poorly understood. We reveal that different group II/III mGluR subtypes show markedly different beta-arrestin (β-arr) coupling and endocytic trafficking. While mGluR2 is resistant to internalization and mGluR3 shows transient β-arr coupling, which enables endocytosis and recycling, mGluR8 and β-arr form stable complexes, which leads to efficient lysosomal targeting and degradation. Using chimeras and mutagenesis, we pinpoint carboxyl-terminal domain regions that control β-arr coupling and trafficking, including the identification of an mGluR8 splice variant with impaired internalization. We then use a battery of high-resolution fluorescence assays to find that heterodimerization further expands the diversity of mGluR regulation. Together, this work provides insight into the relationship between GPCR/β-arr complex formation and trafficking while revealing diversity and intricacy in the regulation of mGluRs.
Seabirds boost coral reef resilience
Authors: Benkwitt Cassandra E. , D’Angelo Cecilia , Dunn Ruth E. , Gunn Rachel L. , Healing Samuel , Mardones M. Loreto , Wiedenmann Joerg , Wilson Shaun K. , Graham Nicholas A. J.
Published: December 2023

Global climate change threatens tropical coral reefs, yet local management can influence resilience. While increasing anthropogenic nutrients reduce coral resistance and recovery, it is unknown how the loss, or restoration, of natural nutrient flows affects reef recovery. Here, we test how natural seabird-derived nutrient subsidies, which are threatened by invasive rats, influence the mechanisms and patterns of reef recovery following an extreme marine heatwave using multiyear field experiments, repeated surveys, and Bayesian modeling. Corals transplanted from rat to seabird islands quickly assimilated seabird-derived nutrients, fully acclimating to new nutrient conditions within 3 years. Increased seabird-derived nutrients, in turn, caused a doubling of coral growth rates both within individuals and across entire reefs. Seabirds were also associated with faster recovery time of
Acropora
coral cover (<4 years) and more dynamic recovery trajectories of entire benthic communities. We conclude that restoring seabird populations and associated nutrient pathways may foster greater coral reef resilience through enhanced growth and recovery rates of corals.
RAD54L2 counters TOP2-DNA adducts to promote genome stability
Authors: D’Alessandro Giuseppina , Morales-Juarez David A. , Richards Sean L. , Nitiss Karin C. , Serrano-Benitez Almudena , Wang Juanjuan , Thomas John C. , Gupta Vipul , Voigt Andrea , Belotserkovskaya Rimma , Goh Chen Gang , Bowden Anne Ramsay , Galanty Yaron , Beli Petra , Nitiss John L. , Zagnoli-Vieira Guido , Jackson Stephen P.
Published: December 2023

The catalytic cycle of topoisomerase 2 (TOP2) enzymes proceeds via a transient DNA double-strand break (DSB) intermediate termed the TOP2 cleavage complex (TOP2cc), in which the TOP2 protein is covalently bound to DNA. Anticancer agents such as etoposide operate by stabilizing TOP2ccs, ultimately generating genotoxic TOP2-DNA protein cross-links that require processing and repair. Here, we identify RAD54 like 2 (RAD54L2) as a factor promoting TOP2cc resolution. We demonstrate that RAD54L2 acts through a novel mechanism together with zinc finger protein associated with tyrosyl-DNA phosphodiesterase 2 (TDP2) and TOP2 (ZATT/ZNF451) and independent of TDP2. Our work suggests a model wherein RAD54L2 recognizes sumoylated TOP2 and, using its ATPase activity, promotes TOP2cc resolution and prevents DSB exposure. These findings suggest RAD54L2-mediated TOP2cc resolution as a potential mechanism for cancer therapy resistance and highlight RAD54L2 as an attractive candidate for drug discovery.
Toward three-dimensional DNA industrial nanorobots
Authors: Zhou Feng , Ni Heng , Zhu Guolong , Bershadsky Lev , Sha Ruojie , Seeman Nadrian C. , Chaikin Paul M.
Published: December 2023

Nanoscale industrial robots have potential as manufacturing platforms and are capable of automatically performing repetitive tasks to handle and produce nanomaterials with consistent precision and accuracy. We demonstrate a DNA industrial nanorobot that fabricates a three-dimensional (3D), optically active chiral structure from optically inactive parts. By making use of externally controlled temperature and ultraviolet (UV) light, our programmable robot, ~100 nanometers in size, grabs different parts, positions and aligns them so that they can be welded, releases the construct, and returns to its original configuration ready for its next operation. Our robot can also self-replicate its 3D structure and functions, surpassing single-step templating (restricted to two dimensions) by using folding to access the third dimension and more degrees of freedom. Our introduction of multiple-axis precise folding and positioning as a tool/technology for nanomanufacturing will open the door to more complex and useful nano- and microdevices.
Intervertebral disc human nucleus pulposus cells associated with back pain trigger neurite outgrowth in vitro and pain behaviors in rats
Authors: Jiang Wensen , Glaeser Juliane D. , Kaneda Giselle , Sheyn Julia , Wechsler Jacob T. , Stephan Stephen , Salehi Khosrowdad , Chan Julie L. , Tawackoli Wafa , Avalos Pablo , Johnson Christopher , Castaneda Chloe , Kanim Linda E.A. , Tanasansomboon Teerachat , Burda Joshua E. , Shelest Oksana , Yameen Haneen , Perry Tiffany G. , Kropf Michael , Cuellar Jason M. , Seliktar Dror , Bae Hyun W. , Stone Laura S. , Sheyn Dmitriy
Published: December 2023

Low back pain (LBP) is often associated with the degeneration of human intervertebral discs (IVDs). However, the pain-inducing mechanism in degenerating discs remains to be elucidated. Here, we identified a subtype of locally residing human nucleus pulposus cells (NPCs), generated by certain conditions in degenerating discs, that was associated with the onset of discogenic back pain. Single-cell transcriptomic analysis of human tissues showed a strong correlation between a specific cell subtype and the pain condition associated with the human degenerated disc, suggesting that they are pain-triggering. The application of IVD degeneration-associated exogenous stimuli to healthy NPCs in vitro recreated a pain-associated phenotype. These stimulated NPCs activated functional human iPSC-derived sensory neuron responses in an in vitro organ-chip model. Injection of stimulated NPCs into the healthy rat IVD induced local inflammatory responses and increased cold sensitivity and mechanical hypersensitivity. Our findings reveal a previously uncharacterized pain-inducing mechanism mediated by NPCs in degenerating IVDs. These findings could aid in the development of NPC-targeted therapeutic strategies for the clinically unmet need to attenuate discogenic LBP.
Zinc prevents vaginal candidiasis by inhibiting expression of an inflammatory fungal protein
Authors: Roselletti Elena , Pericolini Eva , Nore Alexandre , Takacs Peter , Kozma Bence , Sala Arianna , De Seta Francesco , Comar Manola , Usher Jane , Brown Gordon D. , Wilson Duncan
Published: December 2023

Candida
causes an estimated half-billion cases of vulvovaginal candidiasis (VVC) every year. VVC is most commonly caused by
Candida albicans
, which, in this setting, triggers nonprotective neutrophil infiltration, aggressive local inflammation, and symptomatic disease. Despite its prevalence, little is known about the molecular mechanisms underpinning the immunopathology of this fungal infection. In this study, we describe the molecular determinant of VVC immunopathology and a potentially straightforward way to prevent disease. In response to zinc limitation,
C. albicans
releases a trace mineral binding molecule called Pra1 (pH-regulated antigen). Here, we show that the
PRA1
gene is strongly up-regulated during vaginal infections and that its expression positively correlated with proinflammatory cytokine concentrations in women. Genetic deletion of
PRA1
prevented vaginal inflammation in mice, and application of a zinc solution down-regulated expression of the gene and also blocked immunopathology. We also show that treatment of women suffering from recurrent VVC with a zinc gel prevented reinfections. We have therefore identified a key mediator of symptomatic VVC, giving us an opportunity to develop a range of preventative measures for combatting this disease.
A CRISPRi-based genetic resource to study essential
Staphylococcus aureus
genes
Authors: Reed Patricia , Sorg Moritz , Alwardt Dominik , Serra Lúcia , Veiga Helena , Schäper Simon , Pinho Mariana G.
Published: December 2023

We have optimized a clustered regularly interspaced short palindromic repeat (CRISPR) interference system to facilitate gene knockdown in the Gram-positive bacterial pathogen
Staphylococcus aureus
. Our approach used a CRISPRi system derived from
Streptococcus pyogenes,
which involves the co-expression of the
dcas9
gene encoding a catalytically inactive Cas9 protein and a customizable single guide RNA (sgRNA). In our system,
dcas9
is expressed from a single copy in the chromosome of methicillin-resistant
S. aureus
strains COL or JE2, under the control of a tightly regulated promoter, inducible by anhydrotetracycline. The sgRNAs are expressed from a replicative plasmid under the control of a constitutively active promoter. This system enables efficient, inducible, knockdown of both essential and non-essential genes. Using this approach, we constructed the Lisbon CRISPRi Mutant Library comprising 261 strains, in the JE2 background, containing sgRNAs targeting 200 essential genes/operons. This library facilitates the study of the function of essential
S. aureus
genes and is complementary to the Nebraska Transposon Mutant Library, which consists of nearly 2,000 strains, each carrying a transposon insertion within a non-essential gene. The availability of these two libraries will facilitate the study of
S. aureus
pathogenesis and biology.

IMPORTANCE

Staphylococcus aureus
is an important clinical pathogen that causes a high number of antibiotic-resistant infections. The study of
S. aureus
biology, and particularly of the function of essential proteins, is of particular importance to develop new approaches to combat this pathogen. We have optimized a clustered regularly interspaced short palindromic repeat interference (CRISPRi) system that allows efficient targeting of essential
S. aureus
genes. Furthermore, we have used that system to construct a library comprising 261 strains, which allows the depletion of essential proteins encoded by 200 genes/operons. This library, which we have named Lisbon CRISPRi Mutant Library, should facilitate the study of
S. aureus
pathogenesis and biology.
A singular PpaA/AerR-like protein in
Rhodospirillum rubrum
rules beyond the boundaries of photosynthesis in response to the intracellular redox state
Authors: Godoy Manuel S. , de Miguel Santiago R. , Prieto M. Auxiliadora
Published: December 2023

Photosynthesis (PS) in purple non-Sulfur bacteria (PNSB) is strictly controlled to avoid energy consumption and to prevent oxidative stress when oxygen is present. PpsR, a transcriptional repressor encoded in the photosynthesis gene cluster (PGC), is one of the main players in the regulation of PS-related genes. In some cases, PpsR does not sense the environmental cue by itself but delegates this task to other proteins like PpaA/AerR anti-repressors, which can ultimately affect PpsR affinity to some promoter regions. In this work, the effects of locus Rru_A0625 product (HP1) on PS regulation of
Rhodospirillum rubrum
were studied by mutation/complementation and transcriptomic strategies. Rru_A0625 is located next to
ppsR
gene, just like other PpaA/AerR members, and its deletion annuls pigment synthesis in dark micro/anaerobic growth conditions. HP1 shows similarity to PpaA/AerR anti-repressors family, although it does not possess their typical cobalamin binding domain. A transcriptomic analysis of Rru_A0625 deletion mutant showed that HP1 not only has effects on bacteriochlorophyll and carotenoid biosynthesis, but also many other biological processes in the cell. The most notorious is the impact on the transcription of the nitrogenase complex components and accessory proteins. Our results suggest that this new member of the PpaA/AerR family has evolved losing the canonical cobalamin binding domain, but not the redox sensing capability, conserving a not fully understood mechanism of PS regulation.

IMPORTANCE

Rhodospirillum rubrum
vast metabolic versatility places it as a remarkable model bacterium and an excellent biotechnological chassis. The key component of photosynthesis (PS) studied in this work (HP1) stands out among the other members of PpaA/AerR anti-repressor family since it lacks the motif they all share: the cobalamin B-12 binding motif. Despite being reduced and poorly conserved, HP1 stills controls PS as the other members of the family, allowing a fast response to changes in the redox state of the cell. This work also shows that HP1 absence affects genes from relevant biological processes other than PS, including nitrogen fixation and stress response. From a biotechnological perspective, HP1 could be manipulated in approaches where PS is not necessary, such as hydrogen or polyhydroxyalkanoates production, to save energy.
Genome-wide screen reveals cellular functions that counteract rifampicin lethality in
Escherichia coli
Authors: Wang Yu , Fu Han , Shi Xiao-Jie , Zhao Guo-Ping , Lyu Liang-Dong
Published: December 2023

The antibacterial activity of rifamycins specifically relies on the inhibition of transcription by directly binding to the β-subunit of bacterial DNA-dependent RNA polymerase (RNAP). However, its killing efficacy is substantially diminished in most gram-negative bacteria. To systematically reveal the cellular functions that counteract rifamycin-mediated killing in the gram-negative model organism
Escherichia coli
, we performed a genome-wide Tn5 transposon-mediated screen to identify mutants with altered susceptibility to rifampicin. Combined with targeted gene knockouts, our results showed that the β-barrel assembly machinery plays a crucial role in restricting rifampicin from entering the cell, whereas mutants deficient in other cellular permeability barriers, such as lipopolysaccharide and enterobacterial common antigen, had no such effect. At bactericidal concentrations, the killing efficacy of rifampicin was strongly influenced by cellular functions, including iron acquisition, DNA repair, aerobic respiration, and carbon metabolism. Although iron acquisition
de facto
has a strong impact or dependence on cellular redox, our results suggest that their effects on rifampicin efficacy do not rely on hydroxyl radical formation. We provide evidence that maintenance of DNA replication and transcription-coupled nucleotide excision repair protects
E. coli
cells against rifampicin killing. Moreover, our results showed that sustained aerobic respiration and carbon catabolism diminish rifampicin’s killing efficacy, and this effect relies on the inhibition of transcription but not on translation. These findings suggest that the killing efficacy of rifamycins is largely determined by cellular responses upon the inhibition of RNAP and may expand our knowledge of the action mechanisms of rifamycins.

IMPORTANCE

Rifamycins are a group of antibiotics with a wide antibacterial spectrum. Although the binding target of rifamycin has been well characterized, the mechanisms underlying the discrepant killing efficacy between gram-negative and gram-positive bacteria remain poorly understood. Using a high-throughput screen combined with targeted gene knockouts in the gram-negative model organism
Escherichia coli
, we established that rifampicin efficacy is strongly dependent on several cellular pathways, including iron acquisition, DNA repair, aerobic respiration, and carbon metabolism. In addition, we provide evidence that these pathways modulate rifampicin efficacy in a manner distinct from redox-related killing. Our findings provide insights into the mechanism of rifamycin efficacy and may aid in the development of new antimicrobial adjuvants.
Adnexal arteriovenous malformation presenting with spontaneous massive haemoperitoneum
Authors: Chaluvashetty Sreedhara B , Chowhan Prabhjyot Singh , Bhandari Rajan Mani , Shah Jimil
Published: December 2023

Arteriovenous malformations (AVMs) are vascular anomalies composed of a tangle of abnormal vessels in which one or more feeding arteries are directly connected to one or more draining veins via a nidus with no intervening capillary bed. The adnexa are particularly rare sites for the formation of such malformations. Here, we present the case of a middle-aged woman who presented with spontaneous massive haemoperitoneum occurring as a result of a ruptured adnexal AVM. The diagnosis was suspected on transabdominal sonography and confirmed on CT angiography. The patient was shifted to the interventional radiology suite for an urgent angioembolisation following which she improved haemodynamically and her symptoms resolved. The case highlights the fact that although exceedingly rare, gonadal AVMs are an important cause of spontaneous intraperitoneal bleeding. Diagnostic and interventional radiology play an important role in the early and accurate diagnosis of this entity, and angioembolisation can be lifesaving in such patients.
Unrecognised entrapment of a herniated bladder at the pubic symphysis following reduction and external fixation of an unstable pelvic fracture
Authors: Kuo Lu Yu , Silverman Joshua , Morgan Samuel , Steer Richard
Published: December 2023

Bladder injury is frequently associated with complex pelvic fractures with men being the predominant population to sustain such injuries. Entrapment of the bladder through the site of pelvic fracture is a rare clinical entity. We report a case of an entrapped bladder post closed reduction and external fixation of an unstable anteroposterior compression type 3 (APC-3) fracture. This report highlights the diagnostic difficulty with identifying an entrapped bladder and the patient’s functional outcome after 1 year of follow-up.
Trends in the prevalence and social determinants of stunting in India, 2005–2021: findings from three rounds of the National Family Health Survey
Authors: Rao Nirmala , Bala Manya , Ranganathan Namita , Anand Utkarsh , Dhingra Sunaina , Costa Janaina Calu , Weber Ann M
Published: December 2023

Objectives

To assess social determinants of stunting and the shifts in contributions of socio-demographic factors to national prevalence trends in India between 2005 and 2021.

Methods

We leveraged data from three rounds of the National Family Health Survey (NFHS-3: 2005–2006, NFHS-4: 2015–2016, NFHS-5: 2019–2021) for 443 038 children under 5 years. Adjusted logistic regression models and a Kitigawa-Oaxaca-Blinder decomposition were deployed to examine how wealth, residence, belonging to a marginalised social group, maternal education and child sex contributed to changes in stunting prevalence.

Results

The decrease in stunting prevalence was notably slower between NFHS-4 and NFHS-5 (annual average rate of reduction (AARR): 1.33%) than between NFHS-3 and NFHS-4 (AARR: 2.20%). The protective effect of high wealth diminished from 2015 onwards but persisted for high maternal education. However, an intersection of higher household wealth and maternal education mitigated stunting to a greater extent than either factor in isolation. Residence only predicted stunting in 2005–2006 with an urban disadvantage (adjusted OR: 1.18; 95% CI: 1.07 to 1.29). Children from marginalised social groups displayed increased likelihoods of stunting, from 6–16% in 2005–2006 to 11–21% in 2015–2016 and 2020–2021. Being male was associated with 6% and 7% increased odds of stunting in 2015–2016 and 2019–2021, respectively. Increased household wealth (45%) and maternal education (14%) contributed to decreased stunting prevalence between 2005 and 2021.

Conclusions

Stunting prevalence in India has decreased across social groups. However, social disparities in stunting persist and are exacerbated by intersections of low household wealth, maternal education and being from a marginalised social group. Increased survival must be accompanied by needs-based interventions to support children and mitigate mutually reinforcing sources of inequality.