bluuebunny/crossref_metadata_2025 huggingface dataset:
Examining the long‐term effects of COVID‐19 in surgical nurses: Case of Aegean Region
Authors: Yesılyaprak Tugce , Gok Fadime
Published: April 2024

Purpose

To examine the long‐term effects of COVID‐19 on surgical nurses.

Background

Individuals contaminated with COVID‐19 may face several metabolic or psychological issues, primarily in the respiratory, cardiovascular, nervous, musculoskeletal and renal systems during the late period. However, the long‐term epidemiology is still not clear.

Design

Descriptive cross‐sectional study.

Methods

The study included nurses (n = 509) who had been diagnosed with COVID‐19 at least 12 weeks before and worked in surgical departments. We collected the study data via an online survey using the snowball sampling method between December 2021 and May 2022. This study followed the Reporting of Observational Studies in Epidemiology Guideline.

Results

The mean age of the nurses was 31.66 ± 8.74 years. Nurses stated that they were diagnosed with COVID‐19 approximately 36 weeks before participating in this study. We found that the nurses mostly experienced palpitation (83.5%), headache (73.5%), dyspnea (64.1%), anosmia (57.6%), arthralgia (55.7%) and burnout (58.4%) during the late period after COVID‐19.

Conclusion

The long‐term effects of COVID‐19 were related to multiple organ dysfunctions.

No Patient or Public Contribution

Since the study was conducted with healthy individuals who had previously experienced COVID‐19, there is no patient contribution.

Relevance to Clinical Practice

This study focuses on the long‐term effects of COVID‐19 on nurses. The results support the long‐term effects of COVID‐19 and are thought to contribute to the literature.
Role of Piezo1 in modulating the RANKL/OPG ratio in mouse osteoblast cells exposed to Porphyromonas gingivalis lipopolysaccharide and mechanical stress
Authors: Uchinuma Mabuki , Taketani Yoshimasa , Kanaya Risako , Yamane Yusuke , Shiota Koichiro , Suzuki Reiji , Ishii Makiko , Inomata Megumi , Hayashi Joichiro , Shin Kitetsu
Published: April 2024

Aims

Excessive occlusal force with periodontitis leads to rapid alveolar bone resorption. However, the molecular mechanism by which inflammation and mechanical stress cause bone resorption remains unclear. We examined the role of Piezo1, a mechanosensitive ion channel expressed on osteoblasts, in the changes in the receptor activator of nuclear factor‐kappa B ligand (RANKL)/osteoprotegerin (OPG) ratio in mouse MC3T3‐E1 osteoblast‐like cells under Porphyromonas gingivalis lipopolysaccharide (P.g.‐LPS) and mechanical stress.

Methods

To investigate the effect of P.g.‐LPS and mechanical stress on the RANKL/OPG ratio and Piezo1 expression, we stimulated MC3T3‐E1 cells with P.g.‐LPS. After 3 days in culture, shear stress, a form of mechanical stress, was applied to the cells using an orbital shaker. Subsequently, to investigate the role of Piezo1 in the change of RANKL/OPG ratio, we inhibited Piezo1 function by knockdown via Piezo1 siRNA transfection or by adding GsMTx4, a Piezo1 antagonist.

Results

The RANKL/OPG ratio significantly increased in MC3T3‐E1 cells cultured in a medium containing P.g.‐LPS and undergoing mechanical stress compared to cells treated with P.g.‐LPS or mechanical stress alone. However, the expression of Piezo1 was not increased by P.g.‐LPS and mechanical stress. In addition, phosphorylation of MEK/ERK was induced in the cells under P.g.‐LPS and mechanical stress. MC3T3‐E1 cells treated with P.g.‐LPS and mechanical stress when cocultured with RAW264.7 cells induced their differentiation into osteoclast‐like cells. The increased RANKL/OPG ratio was suppressed by either Piezo1 knockdown or the addition of GsMTx4. Furthermore, GsMTx4 inhibited the phosphorylation of MEK/ERK.

Conclusion

These findings suggest that P.g.‐LPS and Piezo1‐mediated mechanical stress induce MEK/ERK phosphorylation and increase RANKL expression in osteoblasts. Consequently, this leads to the differentiation of osteoclast precursor cells into osteoclasts.
Urinalysis and culture results of free‐catch urine samples in dogs: a randomised controlled trial
Authors: Mandese W. W. , Suero M. , Reynolds P. S. , Kariyawasam S. , Beatty S. , Griffin F.
Published: April 2024

Objectives

To evaluate the prevalence of bacterial presence in free‐catch urine samples preceded by either a standardised prepped (“clean‐catch”) protocol versus unprepped (non‐cleaned) voiding.

Materials and Methods

The study was a single‐centre prospective single‐blinded randomised controlled trial. Urine samples were obtained from 100 client‐owned dogs presenting for routine evaluation. Dogs were randomly assigned to either the prepped group (preputial or peri‐vulvar area cleaned with sterile saline before collection) or the unprepped group (no preliminary cleansing) stratified by sex. Urinalysis and urine culture (blood and MacConkey agar) were performed on all samples. Significant bacterial presence on urine culture was defined as >104 colony forming units (CFU)/mL.

Results

There were no statistically significant associations between prepped versus unprepped collection method or sex with a urinalysis positive for bacteriuria. However, on culture, significant bacterial growth was almost five times more likely to be associated with males relative to females (odds ratio 4.59, 95% confidence interval 1.61 to 13.10). The probability of finding a positive culture was not statistically associated with prep method (odds ratio 1.43, 95% confidence interval 0.50 to 4.08).

Clinical Significance

For the majority of dogs without clinical signs of urinary tract infection, free‐catch urine collection does not result in significant bacteriuria found on analysis or culture. The presence of bacteria found in free‐catch samples may be secondary to sample contamination or subclinical bacteriuria. Sample contamination or subclinical bacteriuria may be more prevalent in male dogs.
Recurrent gene flow events occurred during the diversification of clownfishes of the skunk complex
Authors: Marcionetti Anna , Bertrand Joris A. M. , Cortesi Fabio , Donati Giulia F. A. , Heim Sara , Huyghe Filip , Kochzius Marc , Pellissier Loïc , Salamin Nicolas
Published: April 2024

Clownfish (subfamily Amphiprioninae) are an iconic group of coral reef fish that evolved a mutualistic interaction with sea anemones, which triggered the adaptive radiation of the clade. Within clownfishes, the “skunk complex” is particularly interesting. Besides ecological speciation, interspecific gene flow and hybrid speciation are thought to have shaped the evolution of the group. We investigated the mechanisms characterizing the diversification of this complex. By taking advantage of their disjunct geographical distribution, we obtained whole‐genome data of sympatric and allopatric populations of the three main species of the complex (Amphiprion akallopisos, A. perideraion and A. sandaracinos). We examined population structure, genomic divergence and introgression signals and performed demographic modelling to identify the most realistic diversification scenario. We excluded scenarios of strict isolation or hybrid origin of A. sandaracinos. We discovered moderate gene flow from A. perideraion to the ancestor of A. akallopisos + A. sandaracinos and weak gene flow between the species in the Indo‐Australian Archipelago throughout the diversification of the group. We identified introgressed regions in A. sandaracinos and detected in A. perideraion two large regions of high divergence from the two other species. While we found that gene flow has occurred throughout the species' diversification, we also observed that recent admixture was less pervasive than initially thought, suggesting a role of host repartition or behavioural barriers in maintaining the genetic identity of the species in sympatry.
Use of different debriefing methods after in situ simulation with intensive care unit nurses
Authors: Dogu Ozlem , Bozkurt Ramazan , Ziyai Nasibe Yagmur , Elcin Melih , Aygin Dilek
Published: April 2024

Background

The debriefing phase is the simulation phase where performance improves and learning occurs.

Aim

This study examined the effects of the learning conversation (LC)‐based, 3D (defusing, discovering and deepening) model‐based and unstructured debriefing methods on satisfaction and debriefing experience after in‐situ simulation among intensive care unit (ICU) nurses.

Study Design

In this randomized controlled experimental study, three debriefing methods were compared, according to which 119 ICU nurses were divided into the following groups: LC group (n = 38), 3D group (n = 40) and control group (n = 41). In‐ situ simulation was performed with an intensive care patient scenario. p < .05 was considered statistically significant.

Results

The total Debriefing Experience Scale—Experience with Debriefing part scores were 89.76 ± 8.10 in the LC group, 88.90 ± 8.70 in the 3D group and 88.29 ± 7.28 in the control group. No significant difference was found in debriefing experience and satisfaction between the groups (p > .05), but a significant difference was observed in the LC group. The groups showed a homogeneous distribution regarding participant characteristics.

Conclusion

Debriefing experience and satisfaction do not differ between the methods.

Relevance to Clinical Practice

Implementation of the simulation in the ICU in ‐ situ with ICU nurses is beneficial in obtaining a fidelity experience. Performing the debriefing application after simulation in line with the model supports the International Nursing Association for Clinical Simulation and Learning (INACSL) debriefing standards.
Decoupling cell size homeostasis in diatoms from the geometrical constraints of the silica cell wall
Authors: de Haan Diede , Ramos Nahuel‐Hernán , Meng Yu‐Feng , Rotkopf Ron , Addadi Yoseph , Rosenhek‐Goldian Irit , Gal Assaf
Published: April 2024

Unicellular organisms are known to exert tight control over their cell size. In the case of diatoms, abundant eukaryotic microalgae, two opposing notions are widely accepted. On the one hand, the rigid silica cell wall that forms inside the parental cell is thought to enforce geometrical reduction of the cell size. On the other hand, numerous exceptions cast doubt on the generality of this model.

Here, we monitored clonal cultures of the diatom Stephanopyxis turris for up to 2 yr, recording the sizes of thousands of cells, in order to follow the distribution of cell sizes in the population.

Our results show that S. turris cultures above a certain size threshold undergo a gradual size reduction, in accordance with the postulated geometrical driving force. However, once the cell size reaches a lower threshold, it fluctuates around a constant size using the inherent elasticity of cell wall elements.

These results reconcile the disparate observations on cell size regulation in diatoms by showing two distinct behaviors, reduction and homeostasis. The geometrical size reduction is the dominant driving force for large cells, but smaller cells have the flexibility to re‐adjust the size of their new cell walls.
Guard cell and subsidiary cell sizes are key determinants for stomatal kinetics and drought adaptation in cereal crops
Authors: Rui Mengmeng , Chen Rongjia , Jing Yi , Wu Feibo , Chen Zhong‐Hua , Tissue David , Jiang Hangjin , Wang Yizhou
Published: April 2024

Climate change‐induced drought is a major threat to agriculture. C4 crops have a higher water use efficiency (WUE) and better adaptability to drought than C3 crops due to their smaller stomatal morphology and faster response. However, our understanding of stomatal behaviours in both C3 and C4 Poaceae crops is limited by knowledge gaps in physical traits of guard cell (GC) and subsidiary cell (SC).

We employed infrared gas exchange analysis and a stomatal assay to explore the relationship between GC/SC sizes and stomatal kinetics across diverse drought conditions in two C3 (wheat and barley) and three C4 (maize, sorghum and foxtail millet) upland Poaceae crops. Through statistical analyses, we proposed a GCSC‐τ model to demonstrate how morphological differences affect stomatal kinetics in C4 Poaceae crops.

Our findings reveal that morphological variations specifically correlate with stomatal kinetics in C4 Poaceae crops, but not in C3 ones. Subsequent modelling and experimental validation provide further evidence that GC/SC sizes significantly impact stomatal kinetics, which affects stomatal responses to different drought conditions and thereby WUE in C4 Poaceae crops.

These findings emphasize the crucial advantage of GC/SC morphological characteristics and stomatal kinetics for the drought adaptability of C4 Poaceae crops, highlighting their potential as future climate‐resilient crops.
Quantifying the production of plant pollen at the farm scale
Authors: Wright Ellen K. , Timberlake Thomas P. , Baude Mathilde , Vaughan Ian P. , Memmott Jane
Published: April 2024

Plant pollen is rich in protein, sterols and lipids, providing crucial nutrition for many pollinators. However, we know very little about the quantity, quality and timing of pollen availability in real landscapes, limiting our ability to improve food supply for pollinators.

We quantify the floral longevity and pollen production of a whole plant community for the first time, enabling us to calculate daily pollen availability. We combine these data with floral abundance and nectar measures from UK farmland to quantify pollen and nectar production at the landscape scale throughout the year.

Pollen and nectar production were significantly correlated at the floral unit, and landscape level. The species providing the highest quantity of pollen on farmland were Salix spp. (38%), Filipendula ulmaria (14%), Rubus fruticosus (10%) and Taraxacum officinale (9%). Hedgerows were the most pollen‐rich habitats, but permanent pasture provided the majority of pollen at the landscape scale, because of its large area. Pollen and nectar were closely associated in their phenology, with both peaking in late April, before declining steeply in June and remaining low throughout the year.

Our data provide a starting point for including pollen in floral resource assessments and ensuring the nutritional requirements of pollinators are met in farmland landscapes.
Knockdown of circXPO1 inhibits the development of oral squamous cell carcinoma cells
Authors: Lu Yao , Bian Jiaqi , Ferrolino Darwin Omaña , Movahed Fatemeh
Published: April 2024

Background

Circular RNAs (circRNAs) have emerged as pivotal regulators of cellular processes in human malignancies, including oral squamous cell carcinoma (OSCC).

Methods

Reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) was used to detect RNA expression levels of circXPO1, miR‐524‐5p and cyclin D1 (CCND1). Colony formation assay and 5‐Ethynyl‐2′‐deoxyuridine (EdU) assay were performed to analyze cell proliferation, while transwell assay was carried out to investigate the cell migration and invasion. Cell apoptosis was assessed by flow cytometry. Protein expression analysis was implemented by Western blot assay. Additionally, lactate production and glucose consumption were investigated using a lactate assay kit and glucose assay kit, respectively. The in vivo tumorigenic potential of circXPO1 was evaluated using a xenograft mouse model assay.

Results

Elevated levels of circXPO1 and CCND1, alongside reduced miR‐524‐5p expression were decreased in OSCC tissues and cells. Knockdown of circXPO1 in OSCC cells inhibited their proliferative, migratory and invasive capacities, as well as glycolysis, prompting apoptosis. Moreover, circXPO1 silencing hindered tumor growth in vivo. MiR‐524‐5p could be sequestered by circXPO1, and its inhibition could counteract the beneficial effects of circXPO1 knockdown on OSCC progression.

Conclusion

Knockdown of circXPO1 inhibited OSCC progression by up‐regulating miR‐524‐5p and down‐regulating CCND1 expression, which might provide potential targets for OSCC treatment.
Clinical characteristics and salivary biomarkers of burning mouth syndrome
Authors: Kong Mi‐Sun , Kim Moon‐Jong , Kim Yoon‐Young , Chang Ji‐Youn , Kho Hong‐Seop
Published: April 2024

Objectives

To investigate the clinical characteristics and salivary biomarkers in each type of burning mouth syndrome (BMS) patients.

Materials and Methods

Ninety‐eight postmenopausal female patients with BMS were included. Fifty and 21 patients were assigned to the primary and secondary groups, respectively. Twenty‐seven patients with both primary and secondary characteristics were assigned to the intermediate group. Comprehensive clinical characteristics and salivary biomarkers were analyzed.

Results

Significant differences in age, proportion of hyposalivator patients based on unstimulated whole saliva (UWS), symptom distribution, severties of burning sensation and effect of oral complaints in daily life (Eff‐life), and positive symptom distress index (PSDI) were observed among the three groups. The primary group had significant higher UWS flow rate, fewer UWS hyposalivator proportions, and lesser severity of Eff‐life than the secondary group. The intermediate group had significantly greater intensities of burning sensation and Eff‐life and higher PSDI score than did the primary group. The primary group had significantly higher cortisol and dehydroepiandrosterone (DHEA) levels in stimulated whole saliva than did the secondary group.

Conclusions

This study's findings show that clinical characteristics differentiate each BMS type. Cortisol and DHEA levels are potential salivary biomarkers for discriminating between the primary and secondary types of BMS.
Longitudinal dyadic interplay between marital conflict and psychological well‐being in couples: The moderating roles of wives' employment
Authors: Yu Jeong Jin
Published: April 2024

This study examined longitudinal dyadic and within‐partner associations among self‐esteem, subjective happiness, and marital conflict in married couples, with a possible moderating role of wives' employment status. Data were analyzed from the Panel Study on Korean Children, nationwide longitudinal data. The study sample included 1668 married couples (N = 3336 participants) where both partners provided separate data annually across three waves. Husbands' and wives' mean ages at T1 were 39.3 and 36.8 years, respectively. For both partners, self‐esteem and subjective happiness were related bidirectionally at an individual level. Wives' marital conflict was linked to husbands' subsequent marital conflict and vice versa. Wives played a greater role in their husbands’ self‐esteem than vice versa. Findings suggest that wives tend to be their husband's substantial source of perceived psychological well‐being than the reverse; however, the benefit of psychological well‐being is likely to be lower for husbands of stay‐at‐home wives.
A rare complication of pediatric liver transplantation: Post‐transplant diaphragmatic hernia
Authors: Celtik Ulgen , Dokumcu Zafer , Ozcan Coskun , Ergün Orkan
Published: April 2024

Aim

Acquired post‐transplant diaphragmatic hernia (PTDH) is a rare complication of liver transplantation (LT) in children. We aimed to present our experience in PTDH, and a possible causative background is discussed.

Methods

Medical records of patients who had undergone diaphragmatic repair following LT between 2015 and 2023 were reviewed. Demographic information, details of primary diseases necessitating LT, transplantation techniques, and clinical findings associated with PTDH were evaluated.

Results

There were seven patients with PTDH. Median age at transplantation was 69 (range: 9–200) months. Five patients received a left lateral sector, one patient had a right lobe, and one had a left lobe graft. Time between LT and PTDH was 9 (2–123) months. One patient who was diagnosed in the postoperative 10th year was asymptomatic. Respiratory distress and abdominal pain were the main symptoms among all. All patients underwent laparotomy, and primary repair was performed in six patients, and one patient required mesh repair because of a large defect. Small intestine herniated in most cases. There were two complicated cases with perforation of the stomach and colonic volvulus. There is no recurrence or long‐term complications for the median 60 (20–119) month follow‐up period.

Conclusion

PTDH is a rare but serious complication. Majority of symptomatic cases present within the first postoperative year, whereas some late‐presenting cases may not be symptomatic. Inadvertent injury to the inferior phrenic vasculatures due to excessive use of cauterization for control of hemostasis may be a plausible explanation in those cases.
Educational needs of community visiting nurses for infection prevention and control: Application of the Borich needs assessment and the Locus for Focus models
Authors: Kim HyungSeon , Choi Dongwon , Shim Haeun , Sohng Kyeong‐Yae , Choi Min‐Jung
Published: April 2024

Objectives

This study aimed to assess the educational needs and analyze the priorities of infection prevention and control (IPC) for community‐visiting nurses.

Design

This is a cross‐sectional descriptive study.

Sample

This study was conducted with 144 visiting nurses working in public health centers and long‐term care facilities in South Korea.

Method

A total of 23 questions in five subcategories were used to measure the current knowledge and perceived importance of IPC in community‐visiting nursing. Data were collected from June 23 to October 30, 2021, during the COVID‐19 pandemic. Data were analyzed paired t‐test, the Borich needs assessment, and the Locus for Focus models.

Results

Top‐priority content was defined as content belonging to two models, the first 10 contents of Borich needs assessment and the contents located in the Quadrant I of the Locus for Focus models. “Reporting in case of infection‐related accidents,” “Mandatory vaccination for visiting nurses,” “Standard precaution,” “Airborne precaution,” “Contact precautions,” “Respiratory infection control,” and “Post‐visit management.”

Conclusions

This study suggests that it is necessary to provide visiting nurses with more opportunities for IPC education and to develop standardized IPC programs that consider educational priorities.
The effects of photobiomodulation on the improvement of sperm parameters: A review study
Authors: Parvin Ali , Erabi Gisou , Saboohi Tasooji Mohammad Reza , Sadeghpour Sonia , Mellatyar Hassan , Rezaei Arablouydareh Sahar , Navapour Leila , Taheri‐Anganeh Mortaza , Ghasemnejad‐Berenji Hojat
Published: April 2024

The prevalence of male infertility has become a significant clinical concern worldwide, with a noticeable upward trend in recent times. The rates of fertilization and subsequent development of embryos are dependent on many parameters associated with the quality and viability of sperm. Photobiomodulation (PBM) is a promising approach with a great potential for translational applications in the treatment of spermatozoa exhibiting low quality and motility. In this study, a comprehensive analysis of the existing literature, specifically examining the mechanisms of action of PBM has been presented. Our objective was to enhance knowledge in the field of laser light therapy in order to promote the usage of irradiation in clinical settings in a more effective way. Within the realm of reproductive science, the utilization of PBM has been employed to enhance the metabolic processes, motility, and viability of spermatozoa. This is attributed to its advantageous effects on mitochondria, resulting in the activation of the mitochondrial respiratory chain and subsequent synthesis of ATP. This therapeutic approach can be highly advantageous in circumventing the reliance on chemical substances within the culture medium for spermatozoa while also facilitating the viability and motility of spermatozoa, particularly in circumstances involving thawing or samples with significant immotility.
Novel D‐π‐A hemicyanine dye as photoinitiators for in situ hydrogel formation and DLP printing
Authors: Du Yao , Zhang Yating , Liu Shitao , Zhang Xiwang , Wang Tao
Published: April 2024

The field of biofabrication imposes stringent requirements on the polymerization activity and biosafety of photopolymeric hydrogel systems. In this investigation, we designed and synthesized four hemicyanine dyes with a D‐π‐A structure specifically tailored for biofabrication purposes. These novel dyes, incorporating carbazole (CZ), triphenylamine (TPA), anthracene (AN), and benzodithiophene (BDT) as electron donors, along with heterocyclic salt (IN) as electron acceptors, were prepared using a straightforward synthesis method. The absorption maxima of ANIN, CZIN, and TPAIN exceeded 500 nm, rendering them suitable co‐initiators for the free radical photopolymerization of acrylates under green‐red light exposure facilitated by light‐emitting diodes (LEDs) and the co‐initiator iodonium salt (ION). Notably, CZIN and TPAIN, due to their robust dye absorption and efficient electron transfer to ION, functioned as high‐performance photosensitizers. Meanwhile, BDTIN, with its strong and broad absorption range (400–600 nm), enhanced the accuracy of visible light photopolymerization. These dyes exhibit characteristics such as facile synthesis, heightened photo stability, and non‐toxicity and also demonstrate the ability to discern the alkalinity of a solution to some extent. Furthermore, we explored the application of these hemicyanine dyes in 3D printing, showing potential to enhance printing resolution in DLP 3D printing (digital light process 3D printing).
Advocacy coalitions or pragmatic coupling of streams? Explaining policy change in Mexico: The tax reforms of Vicente Fox and Enrique Peña (2001 and 2013)
Authors: Mena Aguilar Oswaldo A.
Published: April 2024

When the former hegemonic PRI won the 2012 election after 12 years of rule by PAN, the President shared with his predecessors a similar institutional setting and divided government. And yet, the PRI managed to pass all items of an extensive 2013 tax reform. If the setting was the same, why did PRI succeed when PAN had only limited success? Why was the 2013 tax reform progressive if PAN and PRI were ideologically proximate? Current explanations that characterize PAN's 2001 tax reforms as a failure because of an unsuccessful change to the Value‐Added Tax Law can accommodate neither the puzzling 2013 extensive tax overhaul nor the changes to the Income Tax Law that were indeed approved in 2001. To navigate these puzzles, I contrast the elements of both proposals and rely on two public policy theoretical frameworks literature, the Advocacy Coalitions and Multiple Streams frameworks to find that, despite representing political compromise over a shared agenda, the outcomes in 2001 and 2013 had less to do with commitment to a core of mutual beliefs and more with policy entrepreneurs of PRI and PRD taking a gamble on a window of opportunity.
Repeated colonization of the Northern Limestone Alps from the Southern Limestone Alps by the rock‐dwelling snail species Cochlostoma henricae
Authors: Hausdorf Bernhard , Xu Jie , Bamberger Sonja
Published: April 2024

Hausdorf, B., Xu, J., BAMBERGER, S. (2024) Repeated colonization of the Northern Limestone Alps from the Southern Limestone Alps by the rock‐dwelling snail species Cochlostoma henricae—Zoologica Scripta, 00, 000–000. The evolution of biogeographic disjunctions in the Southern and Northern Limestone Alps has been studied only in a few taxa. Cochlostoma henricae (Strobel, 1851) is a land snail species restricted to limestone rocks showing such disjunctions. We studied the phylogeny and population structure of this species using genomic ddRAD data to better understand the differentiation of the species in the Southern Alps and the origin of the isolated occurrences in the Northern Limestone Alps. Phylogenetic and population‐genetic analyses of the ddRAD data revealed a subdivision of the C. henricae populations into a western cluster and an eastern cluster. These clusters were probably separated by glaciers that extended from the Piave Valley to the margin of the Alps. Such a longitudinal subdivision is a common pattern of several species in the Southern Limestone Alps. However, the boundaries between western and eastern population groups differ between taxa. Cochlostoma henricae colonized the Northern Limestone Alps at least twice. The population from Breitenwang in Tyrol originated from the western group, whereas the occurrences in the Salzkammergut are from the eastern group. These and similar disjunctions of several other land snails show that even species with limited dispersal abilities could cross the Alps repeatedly by passive long‐distance dispersal events. A subdivision of C. henricae into three subspecies, proposed based on shell characters, does not correspond to its population‐genetic structure. Instead, the eastern and western population groups represent distinct evolutionary units.
Multilocus genetic and morphological phylogenetic analysis: Unveiling a new genus and species in the Tribe Nannenini of jumping spiders (Araneae, Salticidae)
Authors: Satkunanathan Abira , Benjamin Suresh P.
Published: April 2024

Jumping spiders (Salticidae) are a diverse group of non‐web‐building predators and the most species‐rich spider family. The Salticidae Tribe Nannenini consists of a small group of Oriental jumping spiders that are very poorly known. The use of explicit phylogenetic analyses to establish its affinities has so far remained limited. The Nannenini genus Epidelaxia has a peculiar morphology, making its systematic position uncertain. In this study, we present a comprehensive dataset combining molecular and morphological data for Nannenini, including exemplars from all genera. This dataset comprises DNA sequences of approximately 2400 bp, including nuclear genes (18S, 28S and H3) and the mitochondrial gene CO1, sequenced for 40 taxa (20 ingroups and 20 outgroups), along with 61 morphological characters. Notably, both DNA sequences and morphological traits were analysed separately and concurrently for the first time. The Nannenini clade is recovered with high support. Further, Epidelaxia is monophyletic and sister to Tubalaxia gen. n. This is the first hypothesis on the internal phylogenetic structure of Epidelaxia and its placement within the Tribe Nannenini. A new genus Tubalaxia gen. n. and the following new species are described: E. bharathi sp. n., E. somasundaram sp. n., T. castanea sp. n., T. aurea sp. n. The following new combination is proposed: Tubalaxia minuta (Prószyński, 1992) comb. n. Epidelaxia albostellata, E. albocruciata and E. obscura are redescribed. It is worth noting that the newly described species face endangerment due to their limited distribution and small population sizes.
Optical recordings of organellar membrane potentials and the components of membrane conductance in lysosomes
Authors: Castillo‐Velasquez Cristian , Matamala Ella , Becerra Diego , Orio Patricio , Brauchi Sebastian E.
Published: April 2024

The eukaryotic cell is highly compartmentalized with organelles. Owing to their function in transporting metabolites, metabolic intermediates and byproducts of metabolic activity, organelles are important players in the orchestration of cellular function. Recent advances in optical methods for interrogating the different aspects of organellar activity promise to revolutionize our ability to dissect cellular processes with unprecedented detail. The transport activity of organelles is usually coupled to the transport of charged species; therefore, it is not only associated with the metabolic landscape but also entangled with membrane potentials. In this context, the targeted expression of fluorescent probes for interrogating organellar membrane potential (Ψorg) emerges as a powerful approach, offering less‐invasive conditions and technical simplicity to interrogate cellular signalling and metabolism. Different research groups have made remarkable progress in adapting a variety of optical methods for measuring and monitoring Ψorg. These approaches include using potentiometric dyes, genetically encoded voltage indicators, hybrid fluorescence resonance energy transfer sensors and photoinduced electron transfer systems. These studies have provided consistent values for the resting potential of single‐membrane organelles, such as lysosomes, the Golgi and the endoplasmic reticulum. We can foresee the use of dynamic measurements of Ψorg to study fundamental problems in organellar physiology that are linked to serious cellular disorders. Here, we present an overview of the available techniques, a survey of the resting membrane potential of internal membranes and, finally, an open‐source mathematical model useful to interpret and interrogate membrane‐bound structures of small volume by using the lysosome as an example.

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Exploring Gender Diversity and Trauma- and Violence-Informed Sport for Development
Authors: Ferreira Gomes Julia , Hayhurst Lyndsay M.C. , Darroch Francine , Warner Marika
Published: April 2024

The COVID-19 pandemic exacerbated gender-based violence (GBV) rates in Canada, especially affecting young women and gender-diverse youth. Trauma- and violence-informed physical activity and sport for development (SFD) are recognized strategies for supporting survivors of violence, preventing GBV, and challenging gender norms. This paper explores the potential of trauma- and violence-informed approaches with a Canadian SFD organization, focusing on programs aimed to promote gender diversity and address GBV. Grounded in intersectional feminism and queer theory, findings from interviews with SFD staff and participants (n = 15) revealed challenges in maintaining a trauma- and violence-informed approach in a heteronormative sporting space. In conclusion, further research on trauma-informed SFD is crucial due to increasing GBV rates in sport.
EGFR-dependent endocytosis of Wnt9a and Fzd9b promotes β-catenin signaling during hematopoietic stem cell development in zebrafish
Authors: Nguyen Nicole , Carpenter Kelsey A. , Ensing Jessica , Gilliland Carla , Rudisel Emma J. , Mu Emily M. , Thurlow Kate E. , Triche Timothy J. , Grainger Stephanie
Published: April 2024

Cell-to-cell communication through secreted Wnt ligands that bind to members of the Frizzled (Fzd) family of transmembrane receptors is critical for development and homeostasis. Wnt9a signals through Fzd9b, the co-receptor LRP5 or LRP6 (LRP5/6), and the epidermal growth factor receptor (EGFR) to promote early proliferation of zebrafish and human hematopoietic stem cells during development. Here, we developed fluorescently labeled, biologically active Wnt9a and Fzd9b fusion proteins to demonstrate that EGFR-dependent endocytosis of the ligand-receptor complex was required for signaling. In human cells, the Wnt9a-Fzd9b complex was rapidly endocytosed and trafficked through early and late endosomes, lysosomes, and the endoplasmic reticulum. Using small-molecule inhibitors and genetic and knockdown approaches, we found that Wnt9a-Fzd9b endocytosis required EGFR-mediated phosphorylation of the Fzd9b tail, caveolin, and the scaffolding protein EGFR protein substrate 15 (EPS15). LRP5/6 and the downstream signaling component AXIN were required for Wnt9a-Fzd9b signaling but not for endocytosis. Knockdown or loss of EPS15 impaired hematopoietic stem cell development in zebrafish. Other Wnt ligands do not require endocytosis for signaling activity, implying that specific modes of endocytosis and trafficking may represent a method by which Wnt-Fzd specificity is established.
Precise genome editing underlines the distinct contributions of mutations in
ERG11
,
ERG3
,
MRR1
, and
TAC1
genes to antifungal resistance in
Candida parapsilosis
Authors: Hartuis Sophie , Ourliac-Garnier Isabelle , Robert Estelle , Albassier Marjorie , Duchesne Léa , Beaufils Clara , Kuhn Joséphine , Le Pape Patrice , Morio Florent
Published: April 2024

Candida parapsilosis
has recently emerged as a major threat due to the worldwide emergence of fluconazole-resistant strains causing clonal outbreaks in hospitals and poses a therapeutic challenge due to the limited antifungal armamentarium. Here, we used precise genome editing using CRISPR-Cas9 to gain further insights into the contribution of mutations in
ERG11
,
ERG3
,
MRR1
, and
TAC1
genes and the influence of allelic dosage to antifungal resistance in
C. parapsilosis
. Seven of the most common amino acid substitutions previously reported in fluconazole-resistant clinical isolates (including Y132F in
ERG11
) were engineered in two fluconazole-susceptible
C. parapsilosis
lineages (ATCC 22019 and STZ5). Each mutant was then challenged
in vitro
against a large array of antifungals, with a focus on azoles. Any possible change in virulence was also assessed in a
Galleria mellonella
model. We successfully generated a total of 19 different mutants, using CRISPR-Cas9. Except for R398I (
ERG11
), all remaining amino acid substitutions conferred reduced susceptibility to fluconazole. However, the impact on fluconazole
in vitro
susceptibility varied greatly according to the engineered mutation, the stronger impact being noted for G583R acting as a gain-of-function mutation in
MRR1
. Cross-resistance with newer azoles, non-medical azoles, but also non-azole antifungals such as flucytosine, was occasionally noted. Posaconazole and isavuconazole remained the most active
in vitro
. Except for G583R, no fitness cost was associated with the acquisition of fluconazole resistance. We highlight the distinct contributions of amino acid substitutions in
ERG11
,
ERG3
,
MRR1
, and
TAC1
genes to antifungal resistance in
C. parapsilosis
.
Warming alters life-history traits and competition in a phage community
Authors: Greenrod Samuel T. E. , Cazares Daniel , Johnson Serena , Hector Tobias E. , Stevens Emily J. , MacLean R. Craig , King Kayla C.
Published: April 2024

Host-parasite interactions are highly susceptible to changes in temperature due to mismatches in species thermal responses. In nature, parasites often exist in communities, and responses to temperature are expected to vary between host-parasite pairs. Temperature change thus has consequences for both host-parasite dynamics and parasite-parasite interactions. Here, we investigate the impact of warming (37°C, 40°C, and 42°C) on parasite life-history traits and competition using the opportunistic bacterial pathogen
Pseudomonas aeruginosa
(host) and a panel of three genetically diverse lytic bacteriophages (parasites). We show that phages vary in their responses to temperature. While 37°C and 40°C did not have a major effect on phage infectivity, infection by two phages was restricted at 42°C. This outcome was attributed to disruption of different phage life-history traits including host attachment and replication inside hosts. Furthermore, we show that temperature mediates competition between phages by altering their competitiveness. These results highlight phage trait variation across thermal regimes with the potential to drive community dynamics. Our results have important implications for eukaryotic viromes and the design of phage cocktail therapies.

IMPORTANCE

Mammalian hosts often elevate their body temperatures through fevers to restrict the growth of bacterial infections. However, the extent to which fever temperatures affect the communities of phages with the ability to parasitize those bacteria remains unclear. In this study, we investigate the impact of warming across a fever temperature range (37°C, 40°C, and 42°C) on phage life-history traits and competition using a bacterium (host) and bacteriophage (parasite) system. We show that phages vary in their responses to temperature due to disruption of different phage life-history traits. Furthermore, we show that temperature can alter phage competitiveness and shape phage-phage competition outcomes. These results suggest that fever temperatures have the potential to restrict phage infectivity and drive phage community dynamics. We discuss implications for the role of temperature in shaping host-parasite interactions more widely.
Polysaccharide intercellular adhesin and proper phospholipid composition are important for aggregation in
Tetragenococcus halophilus
SL10
Authors: Yanagihara Airi , Matsue Kouta , Kobayashi Kurumi , Wakinaka Takura , Mogi Yoshinobu , Watanabe Jun
Published: April 2024

Aggregating strains of
Tetragenococcus halophilus
tend to be trapped during soy sauce mash-pressing process and are, therefore, critical for clear soy sauce production. However, the precise molecular mechanism involved in
T. halophilus
aggregation remains elusive. In previous studies, we isolated a number of aggregating strains, including
T. halophilus
AB4 and AL1, and showed that a cell surface proteinaceous aggregation factor is responsible for their aggregation phenotype. In the present study, we explored the role of polysaccharide intercellular adhesin (PIA) in aggregate formation in
T. halophilus
SL10, isolated from soy sauce. SL10 exhibited similar aggregation to AB4 and AL1 but formed a non-uniform precipitate with distinctive wrinkles at the bottom of the test tube, unlike AB4 and AL1. Insertion sequence mutations in each gene of the
ica
operon diminished aggregation and PIA production, highlighting the critical role of IcaADBC-mediated PIA production in
T. halophilus
aggregation. Furthermore, two non-aggregating cardiolipin synthase (
cls
) gene mutants with intact
ica
operon did not produce detectable PIA. Phospholipid composition analysis in
cls
mutants revealed a decrease in cardiolipin and an increase in phosphatidylglycerol levels, highlighting the association between phospholipid composition and PIA production. These findings provide evidence for the pivotal role of
cls
in PIA-mediated aggregation and lay the foundation for future studies to understand the intricate networks of the multiple aggregation factors governing microbial aggregation.

IMPORTANCE

Aggregation, commonly observed in various microbes, triggers biofilm formation in pathogenic variants and plays a beneficial role in efficient food production in those used for food production. Here, we showed that
Tetragenococcus halophilus
, a microorganism used in soy sauce fermentation, forms aggregates in a polysaccharide intercellular adhesin (PIA)-mediated manner. Additionally, we unveiled the relationship between phospholipid composition and PIA production. This study provides evidence for the presence of aggregation factors in
T. halophilus
other than the proteinaceous aggregation factor and suggests that further understanding of the coordinated action of these factors may improve clarified soy sauce production.
Development of an automated amplicon-based next-generation sequencing pipeline for rapid detection of bacteria and fungi directly from clinical specimens
Authors: Osborn Lucas J. , Fissel John , Gomez Samantha , Mestas Javier , Flores-Vazquez Jessica , Lee Jaehyeon , Hakimjavadi Hesamedin , Costales Cristina , Dien Bard Jennifer
Published: April 2024

The timely identification of microbial pathogens is essential to guide targeted antimicrobial therapy and ultimately, successful treatment of an infection. However, the yield of standard microbiology testing (SMT) is directly related to the duration of antecedent antimicrobial therapy as SMT culture methods are dependent on the recovery of viable organisms, the fastidious nature of certain pathogens, and other pre-analytic factors. In the last decade, metagenomic next-generation sequencing (mNGS) has been successfully utilized as a diagnostic tool for various applications within the clinical laboratory. However, mNGS is resource, time, and labor-intensive—requiring extensive laborious preliminary benchwork, followed by complex bioinformatic analysis. We aimed to address these shortcomings by developing a largely Automated targeted Metagenomic next-generation sequencing (tmNGS) PipeLine for rapId inFectIous disEase Diagnosis (AMPLIFIED) to detect bacteria and fungi directly from clinical specimens. Therefore, AMPLIFIED may serve as an adjunctive approach to complement SMT. This tmNGS pipeline requires less than 1 hour of hands-on time before sequencing and less than 2 hours of total processing time, including bioinformatic analysis. We performed tmNGS on 50 clinical specimens with concomitant cultures to assess feasibility and performance in the hospital laboratory. Of the 50 specimens, 34 (68%) were from true clinical infections. Specimens from cases of true infection were more often tmNGS positive compared to those from the non-infected group (82.4% vs 43.8%, respectively,
P
= 0.0087). Overall, the clinical sensitivity of AMPLIFIED was 54.6% with 85.7% specificity, equating to 70.6% and 75% negative and positive predictive values, respectively. AMPLIFIED represents a rapid supplementary approach to SMT; the typical time from specimen receipt to identification of potential pathogens by AMPLIFIED is roughly 24 hours which is markedly faster than the days, weeks, and months required to recover bacterial, fungal, and mycobacterial pathogens by culture, respectively.

IMPORTANCE

To our knowledge, this represents the first application of an automated sequencing and bioinformatics pipeline in an exclusively pediatric population. Next-generation sequencing is time-consuming, labor-intensive, and requires experienced personnel; perhaps contributing to hesitancy among clinical laboratories to adopt such a test. Here, we report a strong case for use by removing these barriers through near-total automation of our sequencing pipeline.
Using augmented reality in molecular case studies to enhance biomolecular structure-function explorations in undergraduate classrooms
Authors: Vardar-Ulu Didem , Ragab Saif Eldeen , Agrawal Swati , Dutta Shuchismita
Published: April 2024

Molecular case studies (MCSs) are open educational resources that use a storytelling approach to engage students in biomolecular structure-function explorations, at the interface of biology and chemistry. Although MCSs are developed for a particular target audience with specific learning goals, they are suitable for implementation in multiple disciplinary course contexts. Detailed teaching notes included in the case study help instructors plan and prepare for their implementation in diverse contexts. A newly developed MCS was simultaneously implemented in a biochemistry and a molecular parasitology course at two different institutions. Instructors participating in this cross-institutional and multidisciplinary implementation collaboratively identified the need for quick and effective ways to bridge the gap between the MCS authors’ vision and the implementing instructor’s interpretation of the case-related molecular structure-function discussions. Augmented reality (AR) is an interactive and engaging experience that has been used effectively in teaching molecular sciences. Its accessibility and ease-of-use with smart devices (e.g., phones and tablets) make it an attractive option for expediting and improving both instructor preparation and classroom implementation of MCSs. In this work, we report the incorporation of ready-to-use AR objects as checkpoints in the MCS. Interacting with these AR objects facilitated instructor preparation, reduced students’ cognitive load, and provided clear expectations for their learning. Based on our classroom observations, we propose that the incorporation of AR in MCSs can facilitate its successful implementation, improve the classroom experience for educators and students, and make MCSs more broadly accessible in diverse curricular settings.