bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison between the for‐profit human milk industry and nonprofit human milk banking: Time for regulation?
Authors: Shenker Natalie , Linden Jonathan , Wang Betty , Mackenzie Claudia , Hildebrandt Alex Pueyo , Spears Jacqui , Davis Danielle , Nangia Sushma , Weaver Gillian
Published: October 2023

Human milk (HM) is a highly evolutionary selected, complex biofluid, which provides tailored nutrition, immune system support and developmental cues that are unique to each maternal–infant dyad. In the absence of maternal milk, the World Health Organisation recommends vulnerable infants should be fed with screened donor HM (DHM) from a HM bank (HMB) ideally embedded in local or regional lactation support services. However, demand for HM products has arisen from an increasing awareness of the developmental and health impacts of the early introduction of formula and a lack of prioritisation into government‐funded and nonprofit milk banking and innovation. This survey of global nonprofit milk bank leaders aimed to outline the trends, commonalities and differences between nonprofit and for‐profit HM banking, examine strategies regarding the marketing and placement of products to hospital and public customers and outline the key social, ethical and human rights concerns. The survey captured information from 59 milk bank leaders in 30 countries from every populated continent. In total, five companies are currently trading HM products with several early‐stage private milk companies (PMCs). Products tended to be more expensive from PMC than HMB, milk providers were financially remunerated and lactation support for milk providers and recipients was not a core function of PMCs. Current regulatory frameworks for HM vary widely, with the majority of countries lacking any framework, and most others placing HM within food legislation, which does not include the support and care of milk donors and recipient prioritisation. Regulation as a Medical Product of Human Origin was only in place to prevent the sale of HM in four countries; export and import of HM was banned in two countries. This paper discusses the safety and ethical concerns raised by the commodification of HM and the opportunities policymakers have globally and country‐level to limit the potential for exploitation and the undermining of breastfeeding.
A good egg: An evaluation of a social and behavior change communication campaign to increase egg consumption among children in Rwanda
Authors: Siegal Kim , Wekesa Brendah , Custer Emily , Gatwaza Thierry H. , Uweh Jane , Niyonshuti Marthe
Published: October 2023

Childhood malnutrition, which is endemic in rural areas of low‐income countries, leads to a host of deleterious outcomes such as poor cognitive development, low educational attainment and lower lifetime wages. Promoting the consumption of eggs among young children has emerged as a promising strategy to combat childhood malnutrition, though pathways to scale remain unclear. In this paper, we evaluate the impact of a social and behaviour change communication (SBCC) campaign combined with a program in which rural families purchased chickens on credit (poultry + SBCC; n = 769) relative to an arm in which families only received the poultry intervention (poultry only; n = 750), using a difference‐in‐difference estimation strategy with propensity score matching. The SBCC consisted of radio messages, in‐person training, text message reminders and posters. We found a relatively modest but statistically significant increase in the number of times per week respondents in the poultry + SBCC arm reported feeding eggs to children of 0.28 (p = 0.02) compared to the poultry‐only arm. The increase in egg feeding, however, was more pronounced for boys (0.42, p < 0.01) than for girls (0.14, p = 0.26). In addition, the campaign increased egg feeding more for those who were already feeding eggs to children (0.63, p < 0.01) than those who were not engaging in those practices at baseline (0.26, p < 0.01). However, the difference in these differences was not statistically significant. Future campaigns should ensure higher saturation of messaging and include specific messaging around the importance of feeding girls as well as boys. Campaigns seeking to scale up egg feeding quickly could potentially target the easier‐to‐reach segment of caregivers who already occasionally feed eggs to children though these might not be the neediest group.
Tidal gradients, fine‐scale homing and a potential cryptic ecotype of wild spawning pink salmon (Oncorhynchus gorbuscha)
Authors: May Samuel A. , Shedd Kyle R. , Rand Peter S. , Westley Peter A. H.
Published: October 2023

The homing behaviour of salmon is a remarkable natural phenomenon, critical for shaping the ecology and evolution of populations yet the spatial scale at which it occurs is poorly understood. This study investigated the spatial scale and mechanisms driving homing as depicted by spawning site‐choice behaviour in pink salmon (Oncorhynchus gorbuscha) in Prince William Sound, Alaska. Molecular pedigree analyses of over 30,000 adult spawners in four streams revealed that pink salmon exhibit fine‐scale site fidelity within a stream, returning to within <100 m of their parents. Homing behaviours were driven in part by a salinity gradient between intertidal and freshwater environments, with individuals incubated in freshwater environments more than twice as likely to spawn upstream of tidal influence than those incubated in the intertidal. Our findings challenge the traditional view that pink salmon populations are genetically and phenotypically homogenous due to their short freshwater residency as juveniles and high rates of dispersal as returning adults (i.e. straying). This study has important implications for rates of inbreeding, local adaptation and gene flow within populations, and is particularly relevant to the management of salmon hatcheries, given the high incidence of hatchery‐origin pink salmon, reared in freshwater hatchery environments, that stray into wild populations of Prince William Sound.
Ancient DNA is preserved in fish fossils from tropical lake sediments
Authors: Muschick Moritz , Jemmi Eliane , Lengacher Nicholas , Hänsch Stephanie , Wales Nathan , Kishe Mary A. , Mwaiko Salome , Dieleman Jorunn , Lever Mark Alexander , Salzburger Walter , Verschuren Dirk , Seehausen Ole
Published: October 2023

Tropical freshwater lakes are well known for their high biodiversity, and particularly the East African Great Lakes are renowned for their adaptive radiation of cichlid fishes. While comparative phylogenetic analyses of extant species flocks have revealed patterns and processes of their diversification, little is known about evolutionary trajectories within lineages, the impacts of environmental drivers, or the scope and nature of now‐extinct diversity. Time‐structured palaeodata from geologically young fossil records, such as fossil counts and particularly ancient DNA (aDNA) data, would help fill this large knowledge gap. High ambient temperatures can be detrimental to the preservation of DNA, but refined methodology now allows data generation even from very poorly preserved samples. Here, we show for the first time that fish fossils from tropical lake sediments yield endogenous aDNA. Despite generally low endogenous content and high sample dropout, the application of high‐throughput sequencing and, in some cases, sequence capture allowed taxonomic assignment and phylogenetic placement of 17% of analysed fish fossils to family or tribe level, including remains which are up to 2700 years old or weigh less than 1 mg. The relationship between aDNA degradation and the thermal age of samples is similar to that described for terrestrial samples from cold environments when adjusted for elevated temperature. Success rates and aDNA preservation differed between the investigated lakes Chala, Kivu and Victoria, possibly caused by differences in bottom water oxygenation. Our study demonstrates that the sediment records of tropical lakes can preserve genetic information on rapidly diversifying fish taxa over time scales of millennia.
Peacebuilding Accountability: The United Nations Peacebuilding Fund and Community‐Based Monitoring and Evaluation
Authors: Hancock Landon
Published: October 2023

International peacebuilding as a discourse and practice has expanded rapidly in the nearly three decades since the publication of Boutros Boutros‐Ghali's Agenda for Peace. Alongside the growth of peacebuilding efforts has come the realization that many peacebuilding projects conceived of and sponsored by the international community have failed to meet their own objectives or, more importantly, have failed to be embraced fully by those whom they were supposed to help: the individuals and communities attempting to rebuild their lives in post‐conflict countries. One area that has been under‐examined and—in particular—under‐theorized is the role and impact of funder accountability mechanisms on local ownership, community agency, and peacebuilding success. However, in the development field some work has been done to examine accountability models and to try to develop new ones. As part of a project funded by the Council on Foreign Relations, the author served as a local peacebuilding advisor to the United Nations Peacebuilding Fund's monitoring and evaluation team, where he led an effort to develop a more collaborative evaluation method designed to close accountability loops by including a broader range of actors than that normally considered by current evaluation methodologies. This article offers a scoping analysis of the strengths and weaknesses inherent in the interactions between large international donor agencies and local peacebuilding efforts. It also presents a preview of one agency's determined efforts to bridge those gaps and implement programs and processes designed to promote local agency while supporting transparent accountability.
L‐glutathione 1% promotes neuroprotection of nitrergic neurons and reduces the oxidative stress in the jejunum of rats with Walker‐256‐bearing tumor
Authors: de Oliveira Ana Paula , Perles Juliana Vanessa Colombo Martins , de Souza Sara Raquel Garcia , Sestak Sabrina Silva , da Motta Lima Fabiana Galvão , Almeida Gustavo Henrique Doná Rodrigues , Cicero Lídia Rodrigues , Clebis Naianne Kelly , Guarnier Flávia Alessandra , Blegniski Fernanda Pascoal , Vasconcelos Roseane Carvalho , Araújo Aurigena Antunes , Comar Jurandir Fernando , Moreira Lucas Stafuza , Sehaber‐Sierakowski Camila Cavicchioli , Zanoni Kassio Papi Silva , Zanoni Jacqueline Nelisis
Published: October 2023

Aims

Our main goals were to investigate the effects of L‐glutathione (1%) treatment in Walker‐256 tumor‐bearing rats by analyzing immunoreactive neurons (IR), responsive to the nNOS enzyme and 3‐Nitrotyrosine, in their jejunum myenteric plexus. Moreover, the oxidative state and inflammatory process in these animals were investigated.

Methods

Four experimental groups were utilized: control (C), control treated with L‐glutathione (CGT), Walker‐256 tumor‐bearing rats (TW), and Walker‐256 tumor‐bearing rats treated with L‐glutathione (TWGT). After 14 days of tumor inoculation, the jejunum was collected for immunohistochemical techniques and assessment of oxidative status. Plasma was collected to evaluate oxidative status and measure cytokines.

Results

The TW group exhibited a decrease of reduced glutathione in their jejunum, which was prevented in the L‐glutathione treated TWGT group. TW animals presented pronounced oxidative stress by increasing levels of lipoperoxidation in their jejunum and malondialdehyde in their plasma; however, the L‐glutathione treatment in TWGT group was not able to avoid it. The total antioxidant capacity was altered in groups TW and TWGT, yet the last one had a better index in their plasma. The IL‐10, and TNF‐α levels increased in TWGT animals. The nNOS‐IR neuron density decreased in the jejunum myenteric plexus of the TW group, which was avoided in the TWGT group. The nNOS +3‐Nitrotyrosine neurons quantification did not show significative alterations.

Conclusion

The treatment with L‐glutathione (1%) imposed an important defense to some parameters of oxidative stress induced by TW‐256, leading to neuroprotection to the loss in the nNOS‐IR neuron density.
Clinical and molecular features in a cohort of Middle Eastern patients with epidermolysis bullosa
Authors: Bergson Shir , Daniely Daniel , Bomze David , Mohamad Janan , Malovitski Kiril , Meijers Odile , Briskin Valeria , Bihari Ofer , Malchin Natalia , Israeli Shirli , Mashiah Jacob , Falik‐Zaccai Tzipora , Avitan‐Hersh Emily , Eskin‐Schwartz Marina , Allon‐Shalev Stavit , Sarig Ofer , Sprecher Eli , Samuelov Liat
Published: October 2023

Background

Epidermolysis bullosa (EB) features skin and mucosal fragility due to pathogenic variants in genes encoding components of the cutaneous basement membrane. Based on the level of separation within the dermal‐epidermal junction, EB is sub‐classified into four major types including EB simplex (EBS), junctional EB (JEB), dystrophic EB (DEB), and Kindler EB (KEB) with 16 EB‐associated genes reported to date.

Methods

We ascertained a cohort of 151 EB patients of various Middle Eastern ethnic backgrounds.

Results

The cohort was comprised of EBS (64%, 97/151), DEB (21%, 31/151), JEB (12%, 18/151), and KEB (3%, 5/151). KRT14 and KRT5 variants were most common among EBS patients with 43% (42/97) and 46% (45/97) of EBS patients carrying mutations in either of these two genes, respectively. Truncal involvement was more common in KRT14‐associated EBS as compared to EBS due to KRT5 mutations (p < .05). Mutations in COL17A1 and laminin 332‐encoding genes were identified in 55% (10/18) and 45% (8/18) of JEB patients. Scarring alopecia, caries, and EB nevi were most common among JEB patients carrying COL17A1 mutations as compared to laminin 332‐associated JEB (p < .05). Abnormal nails were evident in most DEB and JEB patients while poikiloderma was exclusively observed in KEB (p < .001).

Conclusions

EB patients of Middle Eastern origin were found to feature specific phenotype–genotype correlations of relevance to the diagnosis and genetic counseling of patients in this region.
Sector attraction and the role of job information: Evidence from a conjoint experiment
Authors: Lee Ivan P. , Jilke Sebastian
Published: October 2023

Are public and private employees different? And is this difference due to the fact that different people are attracted to work for government rather than companies? It has been proposed that individuals with certain characteristics, such as having high levels of risk aversion or public service motivation (PSM), are more likely to self‐select into public service. This study argues that this sector attraction effect depends on the amount of job‐related information available to job seekers at different stages of the job search process. We test our hypotheses using a three‐stage conjoint experimental design. The employment sector only matters to job seekers when little direct information is provided. Once more job‐related information is available to them, it diminishes. This effect is especially strong for people with high levels of PSM. These findings suggest that the sector attraction effect is situational, which has important implications for theory and practice of public sector attraction.
Changes in cycling economy and fractional utilization of V̇O2peak during a 40‐min maximal effort exercise test with acute hypobaric hypoxia corresponding to 2800 m of altitude
Authors: Oberholzer Laura , Aamaas Nikolai Søndersrød , Hallén Jostein
Published: October 2023

Introduction

Peak oxygen uptake (V̇O2peak) declines by ~7% per 1000 m of increasing altitude, whereas exercise performance seems reduced to a lesser extent. For example, 800–10 000 m track and field performances are decreased by 0.4%–2.4% above 1000 m as compared to below and some studies show similar drops in cycling performance. A greater decline in V̇O2peak than in endurance performance with altitude suggests a higher fractional utilization of V̇O2peak (%V̇O2peak). Therefore, we hypothesized that the %V̇O2peak is higher with acute hypoxic exposure than near sea level.

Methods

Sixteen lowlanders (8 women, age: 31 ± 7 years [mean ± SD], body mass: 68 ± 8 kg, V̇O2peak: 60 ± 8 mL min−1 kg−1) underwent cycling testing in a hypobaric hypoxic chamber on 6 test days, three conducted at 300 m and three at 2800 m of acute altitude. At both altitudes, V̇O2peak was determined, and during a 40‐min all out maximal effort time trial (TT), mean power output (MPO) and mean V̇O2 (%V̇O2peak) were assessed.

Results

V̇O2peak decreased by 11.2 ± 3.0% (p < 0.001), while MPO during the TT declined by 10.7 ± 3.1% (p < 0.001) at 2800 m as compared to 300 m. During the TT, %V̇O2peak was higher at altitude, corresponding to 75.9 ± 4.5% at 300 m and 78.8 ± 4.2% at 2800 m (p = 0.011), and cycling economy (mL O2 kJ−1) was poorer (+3.4 ± 2.7%, p < 0.001).

Conclusion

The %V̇O2peak was higher during a cycling TT at 2800 m of altitude than near sea level, while cycling economy was poorer. This resulted in a similar reduction in performance and V̇O2peak. Future studies should address the physiological mechanisms underlying the elevated %V̇O2peak.
Influence of a football match on landing biomechanics and jump performance in female football players
Authors: Miralles‐Iborra Aaron , Elvira Jose L. L. , Del Coso Juan , Hernandez‐Sanchez Sergio , Pino‐Ortega Jose , Moreno‐Pérez Victor
Published: October 2023

This study aimed to assess the acute effect of a competitive football match on jump performance and kinematic parameters during jump landing in semiprofessional female football players. Twenty‐two semiprofessional players (20 ± 3 years) underwent a drop jump task for a posterior video analysis of the landing phase. These measurements were obtained at (1) baseline, (2) after, and (3) 48 h after a competitive football match. A one‐way ANOVA with repeated measures was employed to detect differences over the time. There was a main effect of time for maximal knee flexion angle during drop landing (p = 0.001). In comparison with baseline, maximal knee flexion angle was reduced immediately post‐match and was still reduced 48 h after the match (63.4 ± 8.6 vs 57.0 ± 11.7 vs 48.9 ± 19.1, p ≤ 0.038). There was also a main effect of time for drop jump height (p < 0.001). Drop jump height was reduced immediately post‐match and remained low 48 h after the match in comparison with baseline (27.3 ± 3.6 vs 24.5 ± 2.8 ~ 25.5 ± 3.0 cm, p ≤ 0.002). There was a main effect of time on hip flexion angle during landing (p = 0.001), but the pairwise comparison revealed that this variable was not affected immediately post‐match but was lower 48 h after the match than at baseline (50.1 ± 10.1 ~ 50.8 ± 13.2 vs 38.1 ± 17.8 °, p ≤ 0.005). A competitive football match worsened jump performance and several landing biomechanical parameters in female football players, which were still decreased in comparison with baseline even 48 h after the match.
Clitic placement at the syntax‐phonology interface: A case study of Berber*
Authors: El Hankari Abdelhak
Published: October 2023

Berber1 clitics are argued to follow the main verb but may appear in a position preceding the verb in the presence of a Complementiser, Negation or Tense. However, there are cases involving a subset of these categories yet the clitics still follow the verb. In this article, it is argued that neither syntax nor phonology alone can handle the full range of clitic alternations. Instead, the placement of these clitics is claimed to involve both syntax and phonology. It is argued that these clitics undergo syntactic movement from a lower position to the left of the verb for licensing purposes. However, clitics in some syntactically well‐formed positions may still be ill‐formed due to prosodic considerations. Assuming copy theory of movement, the variable positioning of clitics is attributed to the spell‐out of different copies in the movement chain, thus obviating the need for phonological movement.
A novel HLA‐B*58:141 allele identified by next‐generation sequencing in a potential hematopoietic cell recipient
Authors: Sellyei Joseph , Yan Xiaohong , Qiu Wanglong , Vasilescu Elena‐Rodica , Vlad George
Published: October 2023

We report a novel HLA‐B*58 allele, now named B*58:141, identified by next‐generation sequencing.
Genomic consequences associated with Agrobacterium‐mediated transformation of plants
Authors: Thomson Geoffrey , Dickinson Lauren , Jacob Yannick
Published: October 2023

Attenuated strains of the naturally occurring plant pathogen Agrobacterium tumefaciens can transfer virtually any DNA sequence of interest to model plants and crops. This has made Agrobacterium‐mediated transformation (AMT) one of the most commonly used tools in agricultural biotechnology. Understanding AMT, and its functional consequences, is of fundamental importance given that it sits at the intersection of many fundamental fields of study, including plant‐microbe interactions, DNA repair/genome stability, and epigenetic regulation of gene expression. Despite extensive research and use of AMT over the last 40 years, the extent of genomic disruption associated with integrating exogenous DNA into plant genomes using this method remains underappreciated. However, new technologies like long‐read sequencing make this disruption more apparent, complementing previous findings from multiple research groups that have tackled this question in the past. In this review, we cover progress on the molecular mechanisms involved in Agrobacterium‐mediated DNA integration into plant genomes. We also discuss localized mutations at the site of insertion and describe the structure of these DNA insertions, which can range from single copy insertions to large concatemers, consisting of complex DNA originating from different sources. Finally, we discuss the prevalence of large‐scale genomic rearrangements associated with the integration of DNA during AMT with examples. Understanding the intended and unintended effects of AMT on genome stability is critical to all plant researchers who use this methodology to generate new genetic variants.
Physiological and psychological stress response of blood donors during the blood donation process
Authors: Kaur Arshpreet , Kaur Ravneet , Sood Tanvi , Malhotra Anita , Arun Priti , Mittal Kshitija , Kaur Paramjit , Kaur Gagandeep , Prakash Kiran
Published: October 2023

Background and Objectives

Blood donation can be a potentially stressful event, leading to the activation of an acute stress response. Knowing and identifying potential stressors could help in optimizing the donation experience. The present study aimed to measure the physiological and psychological stress changes before, during and after blood donation.

Materials and Methods

Physiological and psychological stress response was assessed in 70 blood donors. To evaluate physiological stress response, pulse rate, respiratory rate, blood pressure (BP), beat‐to‐beat BP and lead II electrocardiogram were recorded. Baroreflex sensitivity was calculated using the available software. Psychological stress response was assessed using the State–Trait Anxiety Inventory scale.

Results

A significant increase in systolic blood pressure, diastolic blood pressure and mean arterial pressure was observed in the pre‐donation period (p < 0.001). Among the time‐domain parameters, SDSD (standard deviation of differences between adjacent respiratory rate intervals) and RMSSD (root mean square of the successive differences) were significantly lower during the post‐donation period (p < 0.005, p < 0.007, respectively). Among the frequency‐domain parameters, LF nu (relative power of the low‐frequency band in normalized units), HF nu (relative power of the high‐frequency band in normalized units) and LF% (relative power of the low‐frequency band in percentage) were significantly lower before donation compared to during donation (p < 0.001, p < 0.001 and p < 0.012, respectively). LF nu, LF% and LF/HF ratio were also significantly lower during donation compared to after donation (p < 0.05, p < 0.016 and p < 0.042, respectively). Baroreflex sensitivity was also statistically higher during the pre‐donation period. State score was significantly higher among the blood donors during the pre‐donation period.

Conclusion

Physiological and psychological stress is experienced by blood donors during the pre‐donation period. A pre‐donation informative conversation should be carried out with each blood donor and potential stressors should be identified in each.
The uniqueness theorem for Gysin coherent characteristic classes of singular spaces
Authors: Banagl Markus , Wrazidlo Dominik J.
Published: October 2023

We establish a general computational scheme designed for a systematic computation of characteristic classes of singular complex algebraic varieties that satisfy a Gysin axiom in a transverse setup. This scheme is explicitly geometric and of a recursive nature terminating on genera of explicit characteristic subvarieties that we construct. It enables us, for example, to apply intersection theory of Schubert varieties to obtain a uniqueness result for such characteristic classes in the homology of an ambient Grassmannian. Our framework applies in particular to the Goresky–MacPherson ‐class by virtue of the Gysin restriction formula obtained by the first author in previous work. We illustrate our approach for a systematic computation of the ‐class in terms of normally nonsingular expansions in examples of singular Schubert varieties that do not satisfy Poincaré duality over the rationals.
Novel Single Bounded Input Control Synchronization Criterion for a Category of Hyperchaotic and Chaotic Systems in Presence of Uncertainties
Authors: Pallav  , Handa Himesh , Sharma B. B.
Published: October 2023

Dynamical systems that have a chaotic underlying structure have a sensitive dependency on the initial conditions and the values of their parameters. In this piece of work, a straightforward method for solving the synchronization issue in master–slave arrangement for a category of chaotic or hyperchaotic systems, in which perturbations are present in the parameters of the response system, is discussed. The desired control signal is bounded by the initial state when the controller is activated. There is just one control input that is used, and it is derived from Lyapunov's concept of stability. In general, it is tricky to synchronize hyperchaotic or chaotic systems with single controller, and the work turns out to be significantly more complex when the parameters of the slave system are perturbed. The feedback controller using single input that has been constructed makes certain that the state variables of the response system are in synchronization with the state variables that correspond to them in the drive system. In order to attain the desired level of synchronization, the required conditions that must be satisfied to do so have been identified utilizing Lyapunov's stability analysis in a simple manner. In addition, numerical illustrations have been provided in order to support and confirm the theoretical findings of the paper.
A Fully Implicit Enthalpy-Porosity Model for Phase-Change
Authors: de Lemos Marcelo J. S. , Hodierne Anatole J. U.
Published: October 2023

This article proposes a new formulation for a phase change model based on the enthalpy-porosity idea. A general one-energy equation model (1EEM) is extended to deal with the melting and solidification of pure substances and alloys. Before melting and after solidification, solid material is seen as a porous media with low porosity and very small permeability. During phase change, thermal equilibrium in the mushy zone is assumed. Viscous and form drag in the volume-averaged momentum equation are reduced as the temperature rises above the melting point. In the energy equation, latent heat is treated implicitly in the accumulation term instead of explicitly as in most works in the literature. Liquid fraction for the entire field is updated after a new temperature field is calculated. Thermophysical properties are updated with the new liquid fraction field. Governing equations are discretized according to the control-volume method. Algebraic equation sets are relaxed with the Simple Method. Inner iterations make use of the Strong Implicit Procedure. Preliminary results indicate good agreement with the literature for pure substances.
Neo-Natal Castration Leads to Subtle Differences in Porcine Anterior Cruciate Ligament Morphology and Function in Adolescence
Authors: Thompson Jacob D. , Howe Danielle , Griffith Emily H. , Fisher Matthew B.
Published: October 2023

Female adolescent athletes are at a higher risk of tearing their anterior cruciate ligament (ACL) than male counterparts. While most work related to hormones has focused on the effects of estrogen to understand the increased risk of ACL injury, there are other understudied factors, including testosterone. The purpose of this study was to determine how surgical castration in the male porcine model influences ACL size and function across skeletal growth. Thirty-six male Yorkshire crossbreed pigs were raised to 3 (juvenile), 4.5 (early adolescent), and 6 months (adolescent) of age. Animals were either castrated (barrows) within 2 weeks after birth or were left intact (boars). Posteuthanasia, joint and ACL size were assessed via MRI, and biomechanics were assessed via a robotic testing system. Joint size increased throughout age, yet barrows had smaller joints than boars. ACL cross-sectional area (CSA), length, volume, and in situ stiffness increased with age, as did the percent contribution of the ACL anteromedial (AM) bundle to resisting loads. Boar ACL, AM bundle, and PL bundle volumes were 19%, 25%, and 15% larger than barrows across ages. However, ACL CSA, in situ stiffness, and bundle contribution were similar between boars and barrows. The barrows had smaller temporal increases in AM bundle function than boars, but these data were highly variable. Early and sustained loss in testosterone leads to subtle differences in ACL morphology but may not influence measures associated with increased injury risk, such as CSA or bundle forces in response to applied loads.
Pulsed Red and Blue Photobiomodulation for the Treatment of Thigh Contusions and Soft Tissue Injury: A Randomized Controlled Trial
Authors: Wells Aaron , Rigby Justin , Castel Chris , Castel Dawn
Published: October 2023

Context: Contusion and soft tissue injuries are common in sports. Photobiomodultion, light and laser therapy, is an effective aid to increase healing rates and improve function after various injury mechanisms. However, it is unclear how well photobiomodulation improves function after a contusion soft tissue injury. This study aimed to determine the effects of a pulsed red and blue photobiomodulation light patch on muscle function following a human thigh contusion injury. Design: Single-blinded randomized control trial design. Methods: We enrolled 46 healthy participants. Participants completed 5 visits on consecutive days. On the first visit, participants completed a baseline isokinetic quadriceps strength testing protocol at 60°/s and 180°/s. On the second visit, participants were struck in the rectus femoris of the anterior thigh with a tennis ball from a serving machine. Immediately following, participants were treated for 30 minutes with an active or placebo photobiomodulation patch (CareWear light patch system, CareWear Corp). Following the treatment, participants completed the same isokinetic quadriceps strength testing protocol. Participants completed the treatment and isokinetic quadriceps strength test during the following daily visits. We normalized the data by calculating the percent change from baseline. We used a mixed model analysis of covariance, with sex as a covariate, to determine the difference between treatment groups throughout the acute recovery process. Results: We found the active photobiomodulation treatment significantly increased over the placebo group, quadriceps peak torque during the 180°/s test (P = .030), and average power during both the 60°/s (P = .041) and 180°/s (P ≤ .001) assessments. The mean peak torque and average power of 180°/s, at day 4, exceeded the baseline levels by 8.9% and 16.8%, respectively. Conclusions: The red and blue photobiomodulation light patch improved muscle strength and power during the acute healing phase of a human thigh contusion injury model.
Incorporation of hair follicles in 3D bioprinted models of human skin
Authors: Motter Catarino Carolina , Cigaran Schuck Desiree , Dechiario Lexi , Karande Pankaj
Published: October 2023

Current approaches fail to adequately introduce complex adnexal structures such as hair follicles within tissue engineered models of skin. Here, we report on the use of 3D bioprinting to incorporate these structures in engineered skin tissues. Spheroids, induced by printing dermal papilla cells (DPCs) and human umbilical vein cells (HUVECs), were precisely printed within a pregelled dermal layer containing fibroblasts. The resulting tissue developed hair follicle–like structures upon maturation, supported by migration of keratinocytes and melanocytes, and their morphology and composition grossly mimicked that of the native skin tissue. Reconstructed skin models with increased complexity that better mimic native adnexal structures can have a substantial impact on regenerative medicine as grafts and efficacy models to test the safety of chemical compounds.
Molecular basis of ClC-6 function and its impairment in human disease
Authors: Zhang Bing , Zhang Sensen , Polovitskaya Maya M. , Yi Jingbo , Ye Binglu , Li Ruochong , Huang Xueying , Yin Jian , Neuens Sebastian , Balfroid Tom , Soblet Julie , Vens Daphné , Aeby Alec , Li Xiaoling , Cai Jinjin , Song Yingcai , Li Yuanxi , Tartaglia Marco , Li Yang , Jentsch Thomas J. , Yang Maojun , Liu Zhiqiang
Published: October 2023

ClC-6 is a late endosomal voltage-gated chloride-proton exchanger that is predominantly expressed in the nervous system. Mutated forms of ClC-6 are associated with severe neurological disease. However, the mechanistic role of ClC-6 in normal and pathological states remains largely unknown. Here, we present cryo-EM structures of ClC-6 that guided subsequent functional studies. Previously unrecognized ATP binding to cytosolic ClC-6 domains enhanced ion transport activity. Guided by a disease-causing mutation (p.Y553C), we identified an interaction network formed by Y553/F317/T520 as potential hotspot for disease-causing mutations. This was validated by the identification of a patient with a de novo pathogenic variant p.T520A. Extending these findings, we found contacts between intramembrane helices and connecting loops that modulate the voltage dependence of ClC-6 gating and constitute additional candidate regions for disease-associated gain-of-function mutations. Besides providing insights into the structure, function, and regulation of ClC-6, our work correctly predicts hotspots for
CLCN6
mutations in neurodegenerative disorders.
Force redistribution in clathrin-mediated endocytosis revealed by coiled-coil force sensors
Authors: Ren Yuan , Yang Jie , Fujita Barbara , Jin Huaizhou , Zhang Yongli , Berro Julien
Published: October 2023

Forces are central to countless cellular processes, yet in vivo force measurement at the molecular scale remains difficult if not impossible. During clathrin-mediated endocytosis, forces produced by the actin cytoskeleton are transmitted to the plasma membrane by a multiprotein coat for membrane deformation. However, the magnitudes of these forces remain unknown. Here, we present new in vivo force sensors that induce protein condensation under force. We measured the forces on the fission yeast Huntingtin-Interacting Protein 1 Related (HIP1R) homolog End4p, a protein that links the membrane to the actin cytoskeleton. End4p is under ~19-piconewton force near the actin cytoskeleton, ~11 piconewtons near the clathrin lattice, and ~9 piconewtons near the plasma membrane. Our results demonstrate that forces are collected and redistributed across the endocytic machinery.
Ideal plasticity and shape memory of nanolamellar high-entropy alloys
Authors: Chen Shuai , Liu Ping , Pei Qingxiang , Yu Zhi Gen , Aitken Zachary H. , Li Wanghui , Wu Zhaoxuan , Banerjee Rajarshi , Srolovitz David J. , Liaw Peter K. , Zhang Yong-Wei
Published: October 2023

Understanding the relationship among elemental compositions, nanolamellar microstructures, and mechanical properties enables the rational design of high-entropy alloys (HEAs). Here, we construct nanolamellar Al

x

CoCuFeNi HEAs with alternating high– and low–Al concentration layers and explore their mechanical properties using a combination of molecular dynamic simulation and density functional theory calculation. Our results show that the HEAs with nanolamellar structures exhibit ideal plastic behavior during uniaxial tensile loading, a feature not observed in homogeneous HEAs. This remarkable ideal plasticity is attributed to the unique deformation mechanisms of phase transformation coupled with dislocation nucleation and propagation in the high–Al concentration layers and the confinement and slip-blocking effect of the low–Al concentration layers. Unexpectedly, this ideal plasticity is fully reversible upon unloading, leading to a remarkable shape memory effect. Our work highlights the importance of nanolamellar structures in controlling the mechanical and functional properties of HEAs and presents a fascinating route for the design of HEAs for both functional and structural applications.
Integrin α3 promotes T
H
17 cell polarization and extravasation during autoimmune neuroinflammation
Authors: Park Eunchong , Barclay William E. , Barrera Alejandro , Liao Tzu-Chieh , Salzler Harmony R. , Reddy Timothy E. , Shinohara Mari L. , Ciofani Maria
Published: October 2023

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) caused by CNS-infiltrating leukocytes, including T
H
17 cells that are critical mediators of disease pathogenesis. Although targeting leukocyte trafficking is effective in treating autoimmunity, there are currently no therapeutic interventions that specifically block encephalitogenic T
H
17 cell migration. Here, we report integrin α3 as a T
H
17 cell–selective determinant of pathogenicity in experimental autoimmune encephalomyelitis. CNS-infiltrating T
H
17 cells express high integrin α3, and its deletion in CD4
+
T cells or
Il17a
fate-mapped cells attenuated disease severity. Mechanistically, integrin α3 enhanced the immunological synapse formation to promote the polarization and proliferation of T
H
17 cells. Moreover, the transmigration of T
H
17 cells into the CNS was dependent on integrin α3, and integrin α3 deficiency enhanced the retention of CD4
+
T cells in the perivascular space of the blood-brain barrier. Integrin α3–dependent interactions continuously maintain T
H
17 cell identity and effector function. The requirement of integrin α3 in T
H
17 cell pathogenicity suggests integrin α3 as a therapeutic target for MS treatment.
Accumulated metagenomic studies reveal recent migration, whole genome evolution, and undiscovered diversity of orthomyxoviruses
Authors: Dudas Gytis , Batson Joshua
Published: October 2023

Metagenomic studies have uncovered many novel viruses by looking beyond hosts of public health or economic interest. However, the resulting viral genomes are often incomplete, and analyses largely characterize the distribution of viruses over their dynamics. Here, we integrate accumulated data from metagenomic studies to reveal geographic and evolutionary dynamics in a case study of
Orthomyxoviridae
, the RNA virus family that includes influenza virus. First, we use sequences of the orthomyxovirid Wǔhàn mosquito virus 6 to track the migrations of its host. We then look at orthomyxovirus genome evolution, finding gene gain and loss across members of the family, especially in the surface proteins responsible for cell and host tropism. We find that the surface protein of Wǔhàn mosquito virus 6 exhibits accelerated non-synonymous evolution suggestive of antigenic evolution, i.e., vertebrate infection, and belongs to a wider quaranjavirid group bearing highly diverged surface proteins. Finally, we quantify the progress of orthomyxovirus discovery and forecast that many diverged
Orthomyxoviridae
members remain to be found. We argue that continued metagenomic studies will be fruitful for understanding the dynamics, evolution, ecology of viruses, and their hosts, regardless of whether novel viruses are identified or not, as long as study designs allowing for the resolution of complete viral genomes are employed.

IMPORTANCE

The number of known virus species has increased dramatically through metagenomic studies, which search genetic material sampled from a host for non-host genes. Here, we focus on an important viral family that includes influenza viruses, the
Orthomyxoviridae
, with over 100 recently discovered viruses infecting hosts from humans to fish. We find that one virus called Wǔhàn mosquito virus 6, discovered in mosquitoes in China, has spread across the globe very recently. Surface proteins used to enter cells show signs of rapid evolution in Wǔhàn mosquito virus 6 and its relatives which suggests an ability to infect vertebrate animals. We compute the rate at which new orthomyxovirus species discovered add evolutionary history to the tree of life, predict that many viruses remain to be discovered, and discuss what appropriately designed future studies can teach us about how diseases cross between continents and species.
A hybridization target enrichment approach for pathogen genomics
Authors: Sundararaman Balaji , Sylvester Matthew D. , Kozyreva Varvara K. , Berrada Zenda L. , Corbett-Detig Russell B. , Green Richard E.
Published: October 2023

Genomic epidemiology uses pathogens’ whole-genome sequences to understand and manage the spread of infectious diseases. Whole-genome data can be used to monitor outbreaks and cluster formation, identify cross-community transmissions, and characterize drug resistance and immune evasion. Typically, bacteria are cultured from clinical samples to obtain DNA for sequencing to generate whole-genome data. However, culture-independent diagnostic methods are utilized for some fastidious bacteria for better diagnostic yield and rapid pathogen genomics. Whole-genome enrichment (WGE) using targeted DNA sequencing enables direct sequencing of clinical samples without having to culture pathogens. However, the cost of capture probes (“baits”) limits the utility of this method for large-scale genomic epidemiology. We developed a cost-effective method named
C
ircular
N
ucleic acid
E
nrichment
R
eagent
s
ynthesis (CNERs) to generate whole-genome enrichment probes. We demonstrated the method by producing probes for
Mycobacterium tuberculosis
, which we used to enrich
M. tuberculosis
DNA that had been spiked at concentrations as low as 0.01% and 100 genome copies against a human DNA background to 1,225-fold and 4,636-fold. Furthermore, we enriched DNA from different
M. tuberculosis
lineages and
M. bovis
and demonstrated the utility of the WGE-CNERs data for lineage identification and drug-resistance characterization using an established pipeline. The CNERs method for whole-genome enrichment will be a valuable tool for the genomic epidemiology of emerging and difficult-to-grow pathogens.

IMPORTANCE

Emerging infectious diseases require continuous pathogen monitoring. Rapid clinical diagnosis by nucleic acid amplification is limited to a small number of targets and may miss target detection due to new mutations in clinical isolates. Whole-genome sequencing (WGS) identifies genome-wide variations that may be used to determine a pathogen’s drug resistance patterns and phylogenetically characterize isolates to track disease origin and transmission. WGS is typically performed using DNA isolated from cultured clinical isolates. Culturing clinical specimens increases turn-around time and may not be possible for fastidious bacteria. To overcome some of these limitations, direct sequencing of clinical specimens has been attempted using expensive capture probes to enrich the entire genomes of target pathogens. We present a method to produce a cost-effective, time-efficient, and large-scale synthesis of probes for whole-genome enrichment. We envision that our method can be used for direct clinical sequencing of a wide range of microbial pathogens for genomic epidemiology.
Tetravalent SARS-CoV-2 S1 subunit protein vaccination elicits robust humoral and cellular immune responses in SIV-infected rhesus macaque controllers
Authors: Khan Muhammad S. , Kim Eun , Le Hingrat Quentin , Kleinman Adam , Ferrari Alessandro , Sammartino Jose C. , Percivalle Elena , Xu Cuiling , Huang Shaohua , Kenniston Thomas W. , Cassaniti Irene , Baldanti Fausto , Pandrea Ivona , Gambotto Andrea , Apetrei Cristian
Published: October 2023

The coronavirus disease 2019 (COVID-19) pandemic has highlighted the need for safe and effective vaccines to be rapidly developed and distributed worldwide, especially considering the emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants. Protein subunit vaccines have emerged as a promising approach due to their proven safety record and ability to elicit robust immune responses. In this study, we evaluated the immunogenicity and efficacy of an adjuvanted tetravalent S1 subunit protein COVID-19 vaccine candidate composed of the Wuhan, B.1.1.7 variant, B.1.351 variant, and P.1 variant spike proteins in a nonhuman primate model with controlled SIVsab infection. The vaccine candidate induced both humoral and cellular immune responses, with T and B cell responses mainly peaking post boost immunization. The vaccine also elicited neutralizing and cross-reactive antibodies, angiotensin-converting enzyme 2 (ACE2)-blocking antibodies, and T cell responses, including spike-specific CD4
+
T cells. Importantly, the vaccine candidate was able to generate Omicron variant spike-binding and ACE2-blocking antibodies without specifically vaccinating with Omicron, suggesting potential broad protection against emerging variants. The tetravalent composition of the vaccine candidate has significant implications for COVID-19 vaccine development and implementation, providing broad antibody responses against numerous SARS-CoV-2 variants.

IMPORTANCE

The study provides important insights into the immunogenicity and efficacy of a tetravalent protein subunit vaccine candidate against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The vaccine induced both humoral and cellular immune responses in nonhuman primates with controlled SIVagm infection and was able to generate Omicron variant-specific antibodies without specifically vaccinating with Omicron. These findings suggest that the tetravalent composition of the vaccine candidate could provide broad protection against multiple SARS-CoV-2 variants while minimizing the risk of immune escape and the emergence of new variants. Additionally, the use of rhesus macaques with controlled SIVsab infection may better represent vaccine immunogenicity in humans with chronic viral diseases, highlighting the importance of preclinical animal models in vaccine development. Overall, the study provides valuable information for the development and implementation of coronavirus disease 2019 vaccines, particularly for achieving global vaccine equity and addressing emerging variants.
Genome sequence of
Streptomyces
BM cluster phage Frankenweenie
Authors: Cleary Katherine E. , Pelagalli Charles , Cassford Marly , Berry Nathan , Aguas Elizabeth , Kim Brandon , deCarvalho Tagide , Jacobs-Sera Deborah , Caruso Steven M. , Cornely Kathleen
Published: October 2023

Frankenweenie is a newly isolated bacteriophage that infects
Streptomyces scabiei
RL-34. Frankenweenie was discovered in Gaithersburg, MD, and has 366 genes comprising a 200,048-bp genome. Frankenweenie is grouped in cluster BM and is predicted to possess a unique tailspike protein that potentially widens its host range.