bluuebunny/crossref_metadata_2025 huggingface dataset:
The Efficacy of Foot Massage for Pain Relief of Laparoscopic Cholecystectomy: A Meta-analysis Study
Authors: Cheng Yunlan , Zhou Xiaomin , Wang Guiying
Published: April 2023

Background:

Foot massage may have some potential in improving pain relief in patients undergoing laparoscopic cholecystectomy, and this meta-analysis aims to explore the impact of foot massage on pain control after laparoscopic cholecystectomy.

Materials and Methods:

PubMed, EMbase, Web of science, EBSCO, and Cochrane library databases were systematically searched, and we included randomized controlled trials assessing the effect of foot massage on pain control after laparoscopic cholecystectomy.

Results:

Five randomized controlled trials were included in the meta-analysis. Overall, compared with control intervention for laparoscopic cholecystectomy, foot massage was associated with significantly lower pain scores at 60 minutes [mean difference (MD)=−1.19; 95% CI=−2.01 to −0.38; P=0.004), pain scores at 90 minutes (MD=−1.41; 95% CI=−1.73 to −1.10; P<0.00001), pain scores at 120 to 150 minutes (MD=−2.20; 95% CI=−2.49 to −1.90; P<0.00001) and reduced need of additional analgesia (odd ratio=0.04; 95% CI=0.02-0.08; P<0.00001), but demonstrated no obvious effect on pain scores at 10 to 30 minutes (MD=−0.25; 95% CI=−0.79 to 0.29; P=0.37).

Conclusions:

Foot massage benefits to improve pain control after laparoscopic cholecystectomy.
Differential roles of CTP synthetases CTPS1 and CTPS2 in cell proliferation
Authors: Minet Norbert , Boschat Anne-Claire , Lane Rebecca , Laughton David , Beer Philip , Asnagli Hélène , Soudais Claire , Bourne Tim , Fischer Alain , Martin Emmanuel , Latour Sylvain
Published: April 2023

The CTP nucleotide is a key precursor of nucleic acids metabolism essential for DNA replication. De novo CTP production relies on CTP synthetases 1 and 2 (CTPS1 and 2) that catalyze the conversion of UTP into CTP. CTP synthetase activity is high in proliferating cells including cancer cells, however, the respective roles of CTPS1 and CTPS2 in cell proliferation are not known. By inactivation of CTPS1 and/or CTPS2 and complementation experiments, we showed that both CTPS1 and CTPS2 are differentially required for cell proliferation. CTPS1 was more efficient in promoting proliferation than CTPS2, in association with a higher intrinsic enzymatic activity that was more resistant to inhibition by 3-Deaza-uridine, an UTP analog. The contribution of CTPS2 to cell proliferation was modest when CTPS1 was expressed, but essential in absence of CTPS1. Public databases analysis of more than 1,000 inactivated cancer cell lines for CTPS1 or CTPS2 confirmed that cell growth is highly dependent of CTPS1 but less of CTPS2. Therefore, our results demonstrate that CTPS1 is the main contributor to cell proliferation.
Attractant and repellent induce opposing changes in the chemoreceptor four-helix bundle ligand-binding domain
Authors: Guo Lu , Wang Yun-Hao , Cui Rui , Huang Zhou , Hong Yuan , Qian Jia-Wei , Ni Bin , Xu An-Ming , Jiang Cheng-Ying , Zhulin Igor B. , Liu Shuang-Jiang , Li De-Feng
Published: April 2023

Motile bacteria navigate toward favorable conditions and away from unfavorable environments using chemotaxis. Mechanisms of sensing attractants are well understood, however molecular aspects of how bacteria sense repellents have not been established. Here, we identified malate as a repellent recognized by the MCP2021 chemoreceptor in a bacteriumComamonas testosteroniand showed that it binds to the same site as an attractant citrate. Binding determinants for a repellent and an attractant had only minor differences, and a single amino acid substitution in the binding site inverted the response to malate from a repellent to an attractant. We found that malate and citrate affect the oligomerization state of the ligand-binding domain in opposing way. We also observed opposing effects of repellent and attractant binding on the orientation of an alpha helix connecting the sensory domain to the transmembrane helix. We propose a model to illustrate how positive and negative signals are generated and transduced across the membrane and built chimera proteins to illustrate a universal nature of the transmembrane signaling by the repellent.
Novel non hot spot modification in Fks1 ofCandida aurisconfers echinocandin resistance
Authors: Kordalewska Milena , Cancino-Prado Geselle , de Almeida Júnior João Nobrega , Brandão Igor Brasil , de Souza Peral Renata Tigulini , Colombo Arnaldo L. , Perlin David S.
Published: April 2023

We determined echinocandin susceptibility andFKS1genotypes of thirteen clinical isolates ofCandida aurisrecovered from four patients at a tertiary care center in Salvador, Brazil. Three isolates were categorized as echinocandin-resistant and harbored a novelFKS1mutation leading to an amino acid change W691L located downstream from hot-spot 1. When introduced to echinocandin-susceptibleC. aurisstrains by CRISPR/Cas9, Fks1 W691L induced elevated MIC values to all echinocandins (ANF 16-32x; CAS >64x; MCF >64x).
Elucidating the clinical and molecular spectrum ofSMARCC2-associated NDD in a cohort of 65 affected individuals
Authors: Bosch Elisabeth , Popp Bernt , Güse Esther , Skinner Cindy , van der Sluijs Pleuntje J. , Maystadt Isabelle , Pinto Anna Maria , Renieri Alessandra , Bruno Lucia Pia , Granata Stefania , Marcelis Carlo , Baysal Özlem , Hartwich Dewi , Holthöfer Laura , Isidor Bertrand , Cogne Benjamin , Wieczorek Dagmar , Capra Valeria , Scala Marcello , De Marco Patrizia , Ognibene Marzia , Jamra Rami Abou , Platzer Konrad , Carter Lauren B. , Kuismin Outi , van Haeringen Arie , Maroofian Reza , Valenzuela Irene , Cuscò Ivon , Martinez-Agosto Julian A. , Rabani Ahna M. , Mefford Heather C. , Pereira Elaine M. , Close Charlotte , Anyane-Yeboa Kwame , Wagner Mallory , Hannibal Mark C. , Zacher Pia , Thiffault Isabelle , Beunders Gea , Umair Muhammad , Bhola Priya T. , McGinnis Erin , Millichap John , van de Kamp Jiddeke M , Prijoles Eloise J. , Dobson Amy , Shillington Amelle , Graham Brett H. , Garcia Evan-Jacob , Galindo Maureen Kelly , Ropers Fabienne G. , Nibbeling Esther AR , Hubbard Gail , Karimov Catherine , Goj Guido , Bend Renee , Rath Julie , Morrow Michelle M , Millan Francisca , Salpietro Vincenzo , Torella Annalaura , Nigro Vincenzo , Kurki Mitja , Stevenson Roger E , Santen Gijs W.E. , Zweier Markus , Campeau Philippe M. , Severino Mariasavina , Reis André , Accogli Andrea , Vasileiou Georgia
Published: April 2023

PURPOSE

Coffin-Siris and Nicolaides-Baraitser syndromes, are recognisable neurodevelopmental disorders caused by germline variants in BAF complex subunits. TheSMARCC2BAFopathy was recently reported. Herein, we present clinical and molecular data on a large cohort.

METHODS

Clinical symptoms for 41 novel and 24 previously published cases were analyzed using the Human Phenotype Ontology. For genotype-phenotype correlation, molecular data were standardized and grouped into non-truncating and likely gene-disrupting variants (LGD). Missense variant protein expression and BAF subunit interactions were examined using 3D protein modeling, co-immunoprecipitation, and proximity-ligation assays.

RESULTS

Neurodevelopmental delay with intellectual disability, muscular hypotonia and behavioral disorders were the major manifestations. Clinical hallmarks of BAFopathies were rare. Clinical presentation differed significantly, with LGD variants being predominantly inherited and associated with mildly reduced or normal cognitive development, while non-truncating variants were mostlyde novoand presented with severe developmental delay. These distinct manifestations and non-truncating variant clustering in functional domains suggest different pathomechanisms.In vitrotesting showed decreased protein expression for N-terminal missense variants similar to LGD.

CONCLUSION

This study improvedSMARCC2variant classification and identified discernibleSMARCC2-associated phenotypes for LGD and non-truncating variants, which were distinct from other BAFopathies. The pathomechanism of most non-truncating variants has yet to be investigated.
Prescribing differences among older adults with differing health cover and socioeconomic status: a cohort study
Authors: Prendergast Ciaran , Flood Michelle , Murry Logan T. , Clyne Barbara , Fahey Tom , Moriarty Frank
Published: April 2023

Introduction

As health reforms move Ireland from a mixed public-private system toward universal healthcare, it is important to understand variations in prescribing practice for patients with differing health cover and socioeconomic status. This study aims to determine how prescribing patterns for patients aged ≥65 years in primary care in Ireland differ between patients with public and private health cover.

Methods

This was an observational study using anonymised data collected as part of a larger study from 44 general practices in Ireland (2011-2018). Data were extracted from electronic records relating to demographics and prescribing for patients aged ≥65 years. The cohort was divided between those with public health cover (via the General Medical Services (GMS) scheme) and those without. Standardised rates of prescribing were calculated for pre-specified drug classes. We also analysed the number of medications, polypharmacy, and trends over time between groups, using multilevel linear regression adjusting for age and sex.

Results

Overall, 42,456 individuals were included (56% female). Most were covered by the GMS scheme (62%, n=26,490). The rate of prescribing in all medication classes was higher for GMS patients compared to non-GMS patients, with the greatest difference in benzodiazepine anxiolytics. The mean number of unique medications prescribed to GMS patients was 10.9 (SD 5.9), and 8.1 (SD 5.8) for non-GMS patients. The number of unique medications prescribed to both GMS and non-GMS cohorts increased over time. The increase was steeper in the GMS group where the mean number of medications prescribed increased by 0.67 medications/year. The rate of increase was 0.13 (95%CI 0.13, 0.14) medications/year lower for non-GMS patients, a statistically significant difference.

Conclusion

Our study found a significantly larger number of medicines were prescribed to patients with public health cover, compared to those without. Increasing medication burden and polypharmacy among older adults may be accelerated for those of lower socioeconomic status. These findings may inform planning for moves towards universal health care, and this would provide an opportunity to evaluate the effect of expanding entitlement on prescribing and medicines use.
PREVELANCE OF ISCHEMIC AND HEMORRAGHIC STROKE AMONG GERAITIC PATIENTS ADMITTED IN THE PUBLIC TERTIARY CARE HOSPITALS OF PESHAWAR
Authors: Haider Zeeshan , Hussain Sayed Sajid
Published: April 2023

Background

Stroke is a life-threatening medical condition that can result in lifelong brain impairment, complications, and demise. Stroke is the world’s second biggest cause of mortality and could soon overtake as the biggest cause of death globally. It has 2 major pathological types’ i.e. ischemic stroke and hemorrhagic stroke. Hypertension, diabetes mellitus, cardiac diseases, smoking, physical inactivity and age are the risk factors that contribute in the occurrence of a stroke.

Objective

To find out the epidemiological status of stroke types among patients admitted in the public tertiary care hospitals of Peshawar.

Methodology

A descriptive cross-sectional study was carried out to determine the prevalence of stroke types in tertiary care hospitals of Peshawar. The sample size calculated for the research study was 109. Convenience sampling technique was used in this study.

Results

This study was performed among 109 research participants. The most affected were males as 51.13% and females were 45.87%.And the rate of ischemic stroke were 71% while that of hemorrhagic stroke were 28%.

Conclusion

the current research study concluded that majority of the patients had ischemic stroke as compared to the hemorrhagic stroke.
Uncovering the history of recombination and population structure in western Canadian stripe rust populations through mating-type alleles
Authors: Holden Samuel , Bakkeren Guus , Hubensky John , Bamrah Ramandeep , Abbasi Mehrdad , Qutob Dinah , de Graaf Mei-Lan , Kim Sang Hu , Kutcher Hadley R. , McCallum Brent D. , Randhawa Harpinder S. , Iqbal Muhammad , Uloth Keith , Burlakoti Rishi , Brar Gurcharn S.
Published: April 2023

The population structure of crop pathogens such asPuccinia striiformisf. sp.tritici(Pst); the cause of wheat stripe rust, is of interest to researchers looking to understand these pathogens on a molecular level, as well as those with an applied focus such as disease epidemiology. Cereal rusts can reproduce sexually or asexually, and the introduction of novel genetic lineages has the potential to cause serious epidemics such as the one caused by ‘Warrior’ lineage in Europe. In a global context,Pstlineages in Canada were not well-characterized and origin of foreign incursions was not known. We used a whole-genome/transcriptome sequencing approach for the CanadianPstpopulation to identify lineages in a global context, origin, and evidence for foreign incursion. More importantly, for the first time ever, we identified nine alleles of the homeodomain mating-type locus in the worldwidePstpopulation and show that previously identified lineages generally exhibit a single pair of these alleles. In addition, we find only two pheromone receptor alleles. We show that the recent population shift from the ‘PstS1’lineage to the ‘PstS1-related’lineage is also associated with the introduction of a novel mating-type allele (b-3) to the Canadian population. We also show evidence for high levels of mating-type diversity in samples associated with the Himalayan center of diversity forPst, including a single Canadian race previously identified as ‘PstPr’(probable recombinant) which we identify as a foreign incursion from China circa. 2010. These data provide comprehensive details on the population biology of CanadianPstdiversity and mating-type alleles in the globalPstpopulation which can be utilized in testing several research questions and hypotheses around sexuality and parasexuality in rust fungi.
A Trisomy 21 Lung Cell Atlas
Authors: Bhattacharya Soumyaroop , Cherry Caroline , Deutsch Gail ,   Birth Defects Research Laboratory (BDRL), Glass Ian A. , Mariani Thomas J. , Alam Denise Al , Danopoulos Soula
Published: April 2023

Trisomy 21 (T21), resulting in Down Syndrome (DS), is the most prevalent chromosomal abnormality worldwide. While pulmonary disease is a major cause of morbidity and mortality in DS, the ontogeny of pulmonary complications remains poorly understood. We recently demonstrated that T21 lung anomalies, including airway branching and vascular lymphatic abnormalities, are initiated in utero. Here, we aimed to describe molecular changes at the single cell level in prenatal T21 lungs. Our results demonstrate differences in the proportion of cell populations and detail changes in gene expression at the time of initiation of histopathological abnormalities. Notably, we identify shifts in the distribution of alveolar epithelial progenitors, widespread induction of key extracellular matrix molecules in mesenchymal cells and hyper-activation of IFN signaling in endothelial cells. This single cell atlas of T21 lungs greatly expands our understanding of antecedents to pulmonary complications and should facilitate efforts to mitigate respiratory disease in DS.
Evolution of the Telencephalon Anterior-Posterior Patterning by Core Endogenous Network Bifurcation
Authors: Sun Chen , Yao Mengchao , Xiong Ruiqi , Su Yang , Zhu Binglin , Ao Ping
Published: April 2023

How did the complex structure of telencephalon evolve? Existing explanations are based on phenomena and lack the first principle. The Darwinian dynamics and the endogenous network theory established a few years ago provide a mathematical and theoretical framework of a dynamical structure, and a general constitutive structure for theory-experiment coupling, respectively, for answering this question from the first principle perspective. By revisiting a gene network that explains the anterior-posterior patterning of the vertebrate telencephalon, we found that with the increase of the cooperative effect in this network, the fixed points gradually evolve, accompanied by the occurrence of two bifurcations. The dynamic behavior of this network consists with the knowledge obtained from experiments on telencephalon evolution. Furtherly, our work drew an answer quantitatively of how the telencephalon anterior-posterior patterning evolved from the pre-vertebrate chordate to the vertebrate and gave a series of verifiable predictions in a first principle manner.

Figure Abstract
QCT-based computational bone strength assessment updated with MRI-derived ‘hidden’ microporosity
Authors: McPhee Samuel , Kershaw Lucy E , Daniel Carola R , Fernández Marta Peña , Cillán-García Eugenio , Taylor Sarah E , Wolfram Uwe
Published: April 2023

Microdamage accumulated through sustained periods of cyclic loading or single over-loading events contributes to bone fragility through a reduction in stiffness and strength. Monitoring microdamagein vivoremains unattainable by clinical imaging modalities. As such, there are no established computational methods for clinical fracture risk assessment that account for microdamage that existsin vivoat any specific timepoint. We propose a method that combines multiple clinical imaging modalities to identify an indicative surro-gate, which we term ’hidden porosity’, that incorporates pre-existing bone microdamagein vivo. To do so, we use the third metacarpal bone of the equine athlete as an exemplary model for fatigue induced microdamage, which coalesces in the subchondral bone. N=10 metacarpals were scanned by clinical quantitative computed tomography (QCT) and mag-netic resonance imaging (MRI). We used a patch-based similarity method to quantify the signal intensity of a fluid sensitive MRI sequence in bone regions where microdamage coa-lesces. The method generated MRI-derived pseudoCT images which were then used to de-termine a pre-existing damage (Dpex) variable to quantify the proposed surrogate and which we incorporate into a nonlinear constitutive model for bone tissue. The minimum, median, and maximum detectedDpexof 0.059, 0.209, and 0.353 reduced material stiffness by 5.9%, 20.9%, and 35.3% as well as yield stress by 5.9%, 20.3%, and 35.3%. Limb-specific voxel-based finite element meshes were equipped with the updated material model. Lateral and medial condyles of each metacarpal were loaded to simulate physiological joint loading dur-ing gallop. The degree of detectedDpexcorrelated with a relative reduction in both condylar stiffness (p= 0.001, R2> 0.74) and strength (p< 0.001, R2> 0.80). Our results illustrate the complementary value of looking beyond clinical CT, which neglects the inclusion of micro-damage due to partial volume effects. As we use clinically available imaging techniques, our results may aid research beyond the equine model on fracture risk assessment in human diseases such as osteoarthritis, bone cancer, or osteoporosis.
Immunoproteasome deficiency results in accelerated brain aging and epilepsy
Authors: Leister Hanna , Krause Felix F. , Gil Beatriz , Prus Ruslan , Prus Inna , Hellhund-Zingel Anne , Mitra Meghma , Rosa Gerbatin Rogerio Da , Delanty Norman , Beausang Alan , Brett Francesca M. , Farrell Michael A. , Cryen Jane , O’Brien Donncha F. , Henshall David , Helmprobst Frederik , Pagenstecher Axel , Steinhoff Ulrich , Visekruna Alexander , Engel Tobias
Published: April 2023

The immunoproteasome is a central protease complex required for optimal antigen presentation. Immunoproteasome activity is also associated with facilitating degradation of misfolded and oxidized proteins, which prevents cellular stress. While extensively studied during diseases with increasing evidence suggesting a role for the immunoproteasome during pathological conditions including neurodegenerative diseases, this enzyme complex is believed to be mainly inactive in the healthy brain. Here, we show an age-dependent increase in polyubiquitination in the brain of wild-type mice, accompanied with induction of immunoproteasomes, which was most prominent in neurons and microglia. In contrast, mice completely lacking immunoproteasomes (triple-knockout (TKO) mice deficient for LMP2, LMP7 and MECL-1), displayed a strong increase in polyubiquitinated proteins already in the young brain and developed spontaneous epileptic seizures, beginning at the age of 6 months. Injections of kainic acid led to high epilepsy-related mortality of aged TKO mice, confirming increased pathological hyperexcitability states. Notably, the expression of the immunoproteasome was reduced in the brains of patients suffering from epilepsy. In addition, aged TKO mice showed increased anxiety, tau hyperphosphorylation and degeneration of Purkinje cell population with the resulting ataxic symptoms and locomotion alterations. Collectively, our study suggests a critical role for the immunoproteasome in the maintenance of a healthy brain during aging.
Dual targeting of MAPK and PI3K pathways unlocks redifferentiation ofBraf-mutated thyroid cancer organoids
Authors: Lasolle Hélène , Schiavo Andrea , Tourneur Adrien , Gillotay Pierre , da Fonseca Bárbara de Faria , Ceolin Lucieli , Monestier Olivier , Aganahi Benilda , Chomette Laura , Kizys Marina Malta Letro , Haenebalcke Lieven , Pieters Tim , Goossens Steven , Haigh Jody , Detours Vincent , Maia Ana Luiza Silva , Costagliola Sabine , Romitti Mírian
Published: April 2023

Thyroid cancer is the most common endocrine malignancy and several genetic events have been described to promote the development of thyroid carcinogenesis. Besides the effects of specific mutations on thyroid cancer development, the molecular mechanisms controlling tumorigenesis, tumor behavior, and drug resistance are still largely unknown. Cancer organoids have been proposed as a powerful tool to study aspects related to tumor development and progression and appear promising to test individual responses to therapies. Here, using mESC-derived thyroid organoids, we developed a BrafV637E- inducible model able to recapitulate the features of papillary thyroid cancerin vitro. Overexpression of the murine BrafV637Emutation, equivalent to BrafV600Ein humans, rapidly triggers to MAPK activation, cell dedifferentiation, and disruption of follicular organization. BrafV637E-expressing organoids show a transcriptomic signature for p53, focal adhesion, ECM-receptor interactions, EMT, and inflammatory signaling pathways. Finally, PTC-like thyroid organoids were used for drug screening assays. The combination of MAPK and PI3K inhibitors reversedBrafV637Eoncogene-promoted cell dedifferentiation while restoring thyroid follicle organization and functionin vitro. Our results demonstrate that pluripotent stem cells-derived thyroid cancer organoids can mimic tumor development and features while providing an efficient tool for testing novel targeted therapies.
PRMT1 is a critical dependency in clear cell renal cell carcinoma through its role in post-transcriptional regulation of DNA damage response genes
Authors: Walton Joseph , Ng Angel SN , Arevalo Karen , Apostoli Anthony , Meens Jalna , Karamboulas Christina , St-Germain Jonathan , Prinos Panagiotis , Dmytryshyn Julia , Chen Eric , Arrowsmith Cheryl H. , Raught Brian , Ailles Laurie
Published: April 2023

Biallelic inactivation of the Von Hippel-Lindau (VHL) tumor suppressor gene occurs in almost all cases of clear cell renal cell carcinoma (ccRCC) and leads to disrupted oxygen sensing and the upregulation of hypoxia-related genetic programs. Although the loss of VHL appears to be a necessary oncogenic driver event in the majority of ccRCC instances, it is not always a sufficient one. Large genomics studies have revealed that co-deletions ofVHLwith genes involved in chromatin regulation are common and important co-drivers of tumorigenesis. Several conserved evolutionary subtypes have been described clinically and the majority implicate disruptions in epigenetic regulators. It is now clear that impairments in cellular epigenetic mechanisms are important co-drivers of disease and signal a potential vulnerability in the epigenetic network of ccRCC cells relative to their normal counterparts. Using a clinically relevant panel of ccRCC models, we herein sought to exploit this potential vulnerability by screening a library of small molecule inhibitors that target a spectrum of epigenetic regulators. We identified MS023, an inhibitor of type I protein arginine methyltransferases (PRMTs) as an agent with antitumor activity. Using orthogonal genetic technologies, we further validated PRMT1 as the specific critical dependency for cancer growth. Mechanistically, our transcriptomic and functional analyses suggest that MS023 treatment results in substantial impairments to cell cycle and DNA damage repair (DDR) pathways, while spawning an accumulation of DNA damage over time. Our PRMT1 specific proteomics analysis revealed an interactome rich in RNA binding proteins including the specific regulator of DDR mRNA metabolism: the BCLAF1/THRAP3 complex. Further investigation suggests that MS023 treatment may result in impairments to DDR specific mRNA activities including nucleocytoplasmic transport and RNA splicing. Together, our data supports PRMT1 as a compelling target in ccRCC and informs a potential mechanism-based strategy for translational development.
RNAi-mediated knockdown of two orphan G-protein coupled receptors reduces fecundity in the yellow fever mosquitoAedes aegypti
Authors: Keyes-Scott Nia I. , Swade Kyle R. , Allen Lena R. , Vogel Kevin J.
Published: April 2023

G-protein coupled receptors (GPCRs) control numerous physiological processes in insects, including reproduction. While many GPCRs have known ligands, orphan GPCRs do not have identified ligands in which they bind. Advances in genomic sequencing and phylogenetics provide the ability to compare orphan receptor protein sequences to sequences of characterized GPCRs, and thus gain a better understanding of their potential function. Our study investigated two orphan GPCRs, AAEL003647 and AAEL019988, in the yellow fever mosquito,Aedes aegypti. From our phylogenetic analysis, we found that AAEL003647 is orthologous to the SIFamide-2/SMYamide receptor, while AAEL019988 is orthologous to the Trapped in endoderm (Tre1) receptor ofDrosophila melanogaster. Tissue-specific expression analysis revealed that both receptors had highest expression in the ovaries, suggesting they may be important for reproduction. We then used RNA interference (RNAi) to knock down both genes and found a significant reduction in the number of eggs laid per individual female mosquito, suggesting both receptors are important forAe. aegyptireproduction.
3D-Printed LEGO®-inspired Titanium Scaffolds for Patient-Specific Regenerative Medicine
Authors: Lee Seunghun S. , Du Xiaoyu , Smit Thijs , Bissacco Elisa G. , Seiler Daniel , de Wild Michael , Ferguson Stephen J.
Published: April 2023

Despite the recent advances in 3D-printing, it is difficult to fabricate implants that optimally fit a defect size/shape. There are some approaches to resolve this issue, such as patient-specific implant based on CT images, however, it is labor-intensive and costly. Especially in developing countries, affordable treatments are required, while still not excluding these patient groups from manufacturing advances. Here, a SLM 3D-printing strategy was used to fabricate a hierarchical, Assemblable Titanium Scaffold(ATS), which can be manually assembled in any shape or size with ease. A surgeon can create a scaffold that would fit to the defect right before the implantation during the surgery. Additionally, the direct inclusion of micro- and macroporous structures via 3D-printing, as well as a double acid-etched surface treatment(ST) in the ATS, ensure improved nutrient flow and cellular activity. Different structures were designed, 3D-printed and then surface treated for the ST groups. Both individual and stacked ATS have sufficient mechanical properties to withstand physiological loading, and the porous groups resulted enhanced cell proliferation, mineralizaton and osteogenesis compared to non-porous group. Furthermore, successful cell attachment and migration between the assembled ATS were observed. Finally, we demonstrate possible medical applications that reveal the potential of the ATS through assembly.
FSHβ links photoperiodic signalling to seasonal reproduction in Japanese quail
Authors: Majumdar Gaurav , Liddle Timothy A , Stewart Calum , Marshall Christopher J , Bain Maureen , Stevenson Tyler J
Published: April 2023

Annual cycles in daylength provide an initial predictive environmental cue that plants and animals use to time seasonal biology. Seasonal changes in photoperiodic information acts to entrain endogenous programs in physiology to optimize an animal’s fitness. Attempts to identify the neural and molecular substrates of photoperiodic time measurement in birds have, to date, focussed on blunt changes in light exposure during a restricted period of photoinducibility. The objectives of these studies were first to characterise a molecular seasonal clock in Japanese quail and second, to identify the key transcripts involved in endogenously generated interval timing that underlies photosensitivity in birds. We hypothesized that the mediobasal hypothalamus (MBH) provides the neuroendocrine control of photoperiod-induced changes in reproductive physiology, and that the pars distalis of the pituitary gland contains an endogenous internal timer for the short photoperiod dependent development of reproductive photosensitivity. Here we report distinct seasonal waveforms of transcript expression in the MBH, and pituitary gland and discovered the patterns were not synchronized across tissues. Follicle-stimulating hormone-β (FSHβ) expression increased during the simulated spring equinox, prior to photoinduced increases in prolactin, thyrotropin-stimulating hormone-β and testicular growth. Diurnal analyses of transcript expression showed sustained elevated levels ofFSHβ under conditions of the spring equinox, compared to autumnal equinox, short (<12L) and long (>12L) photoperiods.FSHβ expression increased in quail held in non-stimulatory short photoperiod, indicative of the initiation of an endogenously programmed interval timer. These data identify that FSHβ establishes a state of photosensitivity for the external coincidence timing of seasonal physiology. The independent regulation of FSHβ expression provides an alternative pathway through which other supplementary environmental cues, such as temperature, can fine tune seasonal reproductive maturation and involution.
Peptidoglycan fromBacillus anthracis InhibitsHuman Macrophage Efferocytosis in Part by Reducing Cell Surface Expression of MERTK and TIM-3
Authors: Mytych Joshua S , Pan Zijian , Lopez-Davis Charmaine , Redinger Nancy , Lawrence Christina , Ziegler Jadith , Popescu Narcis I. , James Judith A. , Farris A. Darise
Published: April 2023

Bacillus anthracispeptidoglycan (PGN) is a major component of the bacterial cell wall and a key pathogen-associated molecular pattern (PAMP) contributing to anthrax pathology, including organ dysfunction and coagulopathy. Increases in apoptotic lymphocytes are a late-stage feature of anthrax and sepsis, suggesting there is a defect in apoptotic clearance. Here, we tested the hypothesis thatB. anthracisPGN inhibits the capacity of human monocyte-derived macrophages (MΦ) to efferocytose apoptotic cells. Exposure of CD163+CD206+MΦ to PGN for 24h impaired efferocytosis in a manner dependent on human serum opsonins but independent of complement component C3. PGN treatment reduced cell surface expression of the pro-efferocytic signaling receptors MERTK, TYRO3, AXL, integrin αVβ5, CD36 and TIM-3, whereas TIM-1, αVβ3, CD300b, CD300f, STABILIN-1 and STABILIN-2 were unaffected. ADAM17 is a major membrane-bound protease implicated in mediating efferocytotic receptor cleavage. We found multiple ADAM17-mediated substrates increased in PGN-treated supernatant suggesting involvement of membrane-bound proteases. ADAM17 inhibitors TAPI-0 and Marimastat prevented TNF release, indicating effective protease inhibition, and modestly increased cell-surface levels of MerTK and TIM-3 but only partially restored efferocytic capacity by PGN-treated MΦ. We conclude that human serum factors are required for optimal recognition of PGN by human MΦ and thatB. anthracisPGN inhibits efferocytosis in part by reducing cell surface expression of MERTK and TIM-3.
Versatile Computer Aided Design of Freeform DNA Nanostructures and Assemblies
Authors: Pfeifer Wolfgang G. , Huang Chao-Min , Poirier Michael G. , Arya Gaurav , Castro Carlos E.
Published: April 2023

Recent advances in structural DNA nanotechnology have been facilitated by design tools that continue to push the limits of structural complexity while simplifying an often-tedious design process. We recently introduced the software MagicDNA, which enables design of complex 3D DNA assemblies with many components; however, the design of structures with freeform features like vertices or curvature still required iterative design guided by simulation feedback and user intuition. Here, we present an updated design tool, MagicDNA 2.0, that automates the design of freeform 3D geometries, leveraging design models informed by coarse-grained molecular dynamics simulations. Our GUI-based, stepwise design approach integrates a high level of automation with versatile control over assembly and sub-component design parameters. We experimentally validated this approach by fabricating a range of DNA origami assemblies with complex freeform geometries, including a 3D Nozzle, G-clef, and Hilbert and Trifolium curves, confirming excellent agreement between design input, simulation, and structure formation.
Adaptive behavior deficits in individuals with 3q29 deletion syndrome
Authors: Pollak Rebecca M , Lindsey Burrell T , Cubells Joseph F , Klaiman Cheryl , Murphy Melissa M , Saulnier Celine A , Walker Elaine F , White Stormi Pulver , Mulle Jennifer G
Published: April 2023

Background

3q29 deletion syndrome (3q29del) is associated with a significantly increased risk for neurodevelopmental and neuropsychiatric phenotypes. Mild to moderate intellectual disability (ID) is common in this population, and previous work by our team identified substantial deficits in adaptive behavior. However, the full profile of adaptive function in 3q29del has not been described, nor has it been compared to other genomic syndromes associated with elevated risk for neurodevelopmental and neuropsychiatric phenotypes.

Methods

Individuals with 3q29del (n=32, 62.5% male) were evaluated using the Vineland Adaptive Behavior Scales, Third Edition, Comprehensive Parent/Caregiver Form (Vineland-3). We explored the relationship between adaptive behavior and cognitive function, executive function, and neurodevelopmental and neuropsychiatric comorbidities in our 3q29del study sample, and we compared subjects with 3q29del to published data on Fragile X syndrome, 22q11.2 deletion syndrome, and 16p11.2 deletion and duplication syndromes.

Results

Individuals with 3q29del had global deficits in adaptive behavior that were not driven by specific weaknesses in any given domain. Individual neurodevelopmental and neuropsychiatric diagnoses had a small effect on adaptive behavior, and the cumulative number of comorbid diagnoses was significantly negatively associated with Vineland-3 performance. Both cognitive ability and executive function were significantly associated with adaptive behavior, and executive function was a better predictor of Vineland-3 performance than cognitive ability. Finally, the severity of adaptive behavior deficits in 3q29del was distinct from previously published data on comparable genomic disorders.

Conclusions

Individuals with 3q29del have significant deficits in adaptive behavior, affecting all domains assessed by the Vineland-3. Executive function is a better predictor of adaptive behavior than cognitive ability in this population and suggests that interventions targeting executive function may be an effective therapeutic strategy.
Next-Generation Nanopore Sensors for Enhanced Detection of Nanoparticles
Authors: Confederat Samuel , Lee Seungheon , Vang Der , Soulias Dimitrios , Marcuccio Fabio , Peace Timotheus I. , Edwards Martin Andrew , Strobbia Pietro , Samanta Devleena , Wälti Christoph , Actis Paolo
Published: April 2023

Nanopore sensing is a technique based on the Coulter principle to analyze and characterize nanoscale materials with single entity resolution. However, its use in nanoparticle characterization has been constrained by the need to tailor the nanopore aperture size to the size of the analyte, precluding the analysis of heterogenous samples. Additionally, nanopore sensors often require the use of high salt concentrations to improve the signal-to-noise ratio, which further limits their ability to study a wide range of nanoparticles that are unstable at high ionic strength.

Here, we report the development of nanopore sensors enhanced by a polymer electrolyte system, enabling the analysis of heterogenous nanoparticle mixtures at low ionic strength. We present a finite element model to explain the anomalous conductive/resistive pulse signals observed and compare these results with experiments. Furthermore, we demonstrate the wide applicability of the method by characterizing metallic nanospheres of varied sizes, plasmonic nanostars with various degrees of branching, and protein-based spherical nucleic acids with different oligonucleotide loadings.

Our system will complement the toolbox of nanomaterials characterization techniques and will enable real-time optimization workflow for engineering a wide range of nanomaterials.
The adaptability of the ion binding site by the Ag(I)/Cu(I) periplasmic chaperone SilF
Authors: Lithgo Ryan M. , Hanževački Marko , Harris Gemma , Kamps Jos J. A. G. , Holden Ellie , Benesch Justin LP , Jäger Christof M. , Croft Anna K. , Hobman Jon L. , Orville Allen M. , Quigley Andrew , Carr Stephen B. , Scott David J.
Published: April 2023

The periplasmic chaperone SilF has been identified as part of an Ag(I) detoxification system in Gram negative bacteria. Sil proteins also bind Cu(I), but with reported weaker affinity, therefore leading to the designation of a specific detoxification system for Ag(I). Using isothermal titration calorimetry we show that binding of both ions is not only tighter than previously thought, but of very similar affinities. We investigated the structural origins of ion binding using molecular dynamics and QM/MM simulations underpinned by structural and biophysical experiments. The results of this analysis showed that the binding site adapts to accommodate either ion, with key interactions with the solvent in the case of Cu(I). The implications of this are that Gram negative bacteria do not appear to have evolved a specific Ag(I) efflux system but take advantage of the existing Cu(I) detoxification system. Therefore, there are consequences for how we define a particular metal resistance mechanism and understand its evolution in the environment.
Polygenic predictions of occupational status GWAS elucidate genetic and environmental interplay for intergenerational status transmission, careers, and health
Authors: Akimova Evelina T. , Wolfram Tobias , Ding Xuejie , Tropf Felix C. , Mills Melinda C.
Published: April 2023

Socioeconomic status (SES) impacts health and the life course. This GWAS on sociologically informed occupational status measures (ISEI, SIOPS, and CAMSIS) using the UKBiobank (N=273,157) identified 106 genetic variants of which 8 are novel to the study of SES. Genetic correlation analyses point to a common genetic factor for SES. Within-family prediction and its reduction was attributable in equal parts to genetic nurture and assortative mating. Using polygenic scores from population predictions of 5-8%, we, firstly, showed that cognitive and non-cognitive traits – including scholastic and occupational motivation and aspiration – link genetic scores to occupational status. Second, 62% of the intergenerational transmission of occupational status can be ascribed to non-genetic inheritance (e.g., family environment). Third, the link between genetics, occupation, and health are interrelated with parental occupational status confounding the genetic prediction of general health. Finally, across careers, genetic prediction compresses during mid-career with divergence in status at later stages.
High-affinity biomolecular interactions are modulated by low-affinity binders
Authors: Mukundan S. , Deshpande Girish , Madhusudhan M.S.
Published: April 2023

Molecular interactions play a central role in all biological processes. The strength of these interactions is often characterized by their dissociation constants (KD). The high affinity interactions (KD≤ 10-8M) are crucial for the proper execution of cellular processes and are thus extensively investigated. Detailed molecular and biochemical analyses of such high affinity interactions have lent considerable support to the concept of binary on/off switches in different biological contexts. However, such studies have typically discounted the presence of low-affinity binders (KD> 10-5M) in the cellular environment. In this study, we have assessed the potential influence of such low affinity binders on high affinity interactions. By employing Gillespie stochastic simulations as well as continuous methods, we demonstrate that the presence of low-affinity binders can indeed alter the kinetics and the steady state of high-affinity interactions. We refer to this effect as ‘herd regulation’ and have evaluated its possible impact in two different contexts including sex determination inDrosophila melanogasterand in signalling systems that employ molecular thresholds. Lastly, we suggest an ‘in vitro’ experimental strategy to validate herd regulation. Based on these analyses, we propose that low-affinity binders are likely prevalent in different biological contexts where the outcomes are contingent upon threshold value determinants and thus potentially impact their homeostatic regulation.
Synthetic lethal interaction between WEE1 and PKMYT1 is a target for multiple low dose treatment of high-grade serous ovarian carcinoma
Authors: Benada Jan , Bulanova Daria , Azzoni Violette , Petrosius Valdemaras , Ghazanfar Saba , Wennerberg Krister , Sørensen Claus Storgaard
Published: April 2023

Ovarian cancer is driven by genetic alterations that necessitate protective DNA damage and replication stress responses through cell cycle control and genome maintenance. This creates specific vulnerabilities that may be exploited therapeutically. WEE1 kinase is a key cell cycle control kinase, and it has emerged as a promising cancer therapy target. However, adverse effects have limited its clinical progress, especially, when tested in combination with chemotherapies. A strong genetic interaction between WEE1 and PKMYT1 led us to hypothesize that a multiple low dose approach utilising joint WEE1 and PKMYT1 inhibition would allow exploitation of the synthetic lethality. In the present study, we found that the combination of WEE1 and PKMYT1 inhibition exhibited synergistic effects in eradicating ovarian cancer cells and ovarian organoid models at a low dose. The WEE1 and PKMYT1 inhibition synergistically promoted activation of CDK1 by decreasing the phosphorylation levels of T14 and Y15 residues. Furthermore, the combined treatment exacerbated DNA replication stress and replication catastrophe, leading to increase of the genomic instability and inflammatory STAT1 signalling activation. Finally, the multiple low dosing was well tolerated in mice. These findings suggest a new multiple low dose approach to harness the potency of WEE1 inhibition through the synthetic lethal interaction with PKMYT1 that may contribute to the development of new treatments for ovarian cancer.
A comprehensive transcriptional signature in pancreatic ductal adenocarcinoma reveals new insights into the immune and desmoplastic microenvironment
Authors: Pérez-Díez Irene , Andreu Zoraida , Hidalgo Marta R. , Perpiñá-Clérigues Carla , Fantín Lucía , Fernandez-Serra Antonio , de la Iglesia-Vaya María , Lopez-Guerrero José A , García-García Francisco
Published: April 2023

Pancreatic ductal adenocarcinoma (PDAC) prognosis and treatment response remains devastatingly poor due partly to the highly heterogeneous, aggressive, and immunosuppressive nature of this tumor type. The intricate relationship between stroma, inflammation, and immunity remains vaguely understood in the PDAC microenvironment. Here, we performed a meta-analysis of stroma-, and immune-related gene expression in the PDAC microenvironment to improve disease prognosis and therapeutic development. We selected twenty-one PDAC studies from the Gene Expression Omnibus and ArrayExpress databases, including 922 samples (320 controls and 602 cases). Differential gene enrichment analysis identified 1153 significant dysregulated genes in PDAC patients that contribute to a desmoplastic stroma and an immunosuppressive environment (the hallmarks of PDAC tumors). The results highlighted two gene signatures related to the immune and stromal environments that cluster PDAC patients in high- and low-risk groups, impacting patient stratification and therapeutic decision-making. Moreover,HCP5, SLFN13, IRF9, IFIT2, andIFI35 immune geneswere related to prognosis value in PDAC patients, for the first time.

Simple Summary

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease with few curative options. Desmoplastic stroma and immune system evasion in PDAC represent challenges to the success of therapeutic strategies that function well in other tumor types. Characterizing the PDAC microenvironment (including the immune environment) remains critical to developing safe and efficient therapies. Here, we present a comprehensive meta-analysis identifying 1153 significantly dysregulated genes, which mainly impact extracellular matrix remodeling and the immune system. We identify two signatures of twenty-eight immune-related genes and eleven stroma-related genes influencing PDAC patient survival. Additionally, five immune genes are associated with PDAC prognosis for the first time.
Induced neural phase precession through exogeneous electric fields
Authors: Wischnewski M. , Tran H. , Zhao Z. , Shirinpour S. , Haigh Z.J. , Rotteveel J. , Perera N.D. , Alekseichuk I , Zimmermann J. , Opitz A.
Published: April 2023

The gradual shifting of preferred neural spiking relative to local field potentials (LFPs), known as phase precession, plays a prominent role in neural coding. Correlations between the phase precession and behavior have been observed throughout various brain regions. As such, phase precession is suggested to be a global neural mechanism that promotes local neuroplasticity. However, causal evidence and neuroplastic mechanisms of phase precession are lacking so far. Here we show a causal link between LFP dynamics and phase precession. In three experiments, we modulated LFPs in humans, a non-human primate, and computational models using alternating current stimulation. We show that continuous stimulation of motor cortex oscillations in humans lead to a gradual phase shift of maximal corticospinal excitability by ∼90°. Further, exogenous alternating current stimulation induced phase precession in a subset of entrained neurons (∼30%) in the non-human primate. Multiscale modeling of realistic neural circuits suggests that alternating current stimulation-induced phase precession is driven by NMDA-mediated synaptic plasticity. Altogether, the three experiments provide mechanistic and causal evidence for phase precession as a global neocortical process. Alternating current-induced phase precession and consequently synaptic plasticity is crucial for the development of novel therapeutic neuromodulation methods.