bluuebunny/crossref_metadata_2025 huggingface dataset:
Addressing Confusion and Inconsistencies Surrounding the Provider Role in Medical Cannabis Programs
Authors: Tang Yi-Lang , McCord Elizabeth , Earley Paul H. , Drexler Karen
Published: July 2024

More than 40 states in the United States have established medical cannabis programs that authorize the use of cannabis for specific medical conditions. Initially, these medical cannabis programs aimed to offer compassionate care primarily for terminal or rare, untreatable conditions. However, the scope of these programs has broadened to include nonterminal and more common conditions, including various medical and mental disorders. This expansion introduces several health care challenges: a lack of robust research evidence for many listed conditions, a sense of unpreparedness among providers, and a disparity in provider roles, expectations, and responsibilities across different states, leading to potential confusion. To address these issues, medical organizations need to develop expert consensus or guidelines that underscore evidence-based shared decision-making and patient monitoring standards. Medical education should also include such training. Concurrently, medical providers must prioritize evidence-based treatment over public opinion, exercise clinical judgment, and take responsibility for their recommendations.
Preliminary Study of Motion Preservation Following Posterior Dynamic Distraction Device in Adolescent Idiopathic Scoliosis Patients
Authors: Todderud Julia E. , Milbrandt Todd A. , Floyd Edward , Haft Geoffrey , El-Hawary Ron , Albert Michael , Larson A. Noelle
Published: July 2024

Background:

Motion-sparing scoliosis surgeries such as the posterior dynamic distraction device (PDDD) are slowly increasing in use. However, there is limited clinical data documenting postoperative motion across the PDDD construct. With this cohort study, we aim to measure sagittal and coronal motion following PDDD. We hypothesize coronal and sagittal spinal motion will be partially preserved across the construct.

Methods:

Retrospective review of prospectively collected data. Preoperative and minimum 1-year postoperative coronal range of motion across the instrumented levels was compared. Available flexion/extension radiographs were evaluated postoperatively to assess sagittal arc of motion. Radiographs from latest follow-up were used.

Results:

At a mean of 1.9 years (1 to 5 y), flexibility radiographs were available on 29 patients treated with PDDD (17 thoracic, 12 lumbar). Mean age at surgery was 16 years (12 to 25). Postoperative coronal arc of motion in PDDD patients was 11 degrees (3 to 19 degrees) in the thoracic spine and 10 degrees (0 to 28 degrees) in the lumbar spine. Compared with preoperative motion, the thoracic arc of motion was maintained by 33% (35 to 11 degrees) and lumbar motion was maintained by 30% (34 to 10 degrees). Flexion-extension radiographs were available on 7 patients. Sagittal arc for the upper instrumented vertebral end plate to the lower instrumented vertebral endplate of the cohort was 10 degrees in the thoracic spine (6 to 18) and 14 degrees in the lumbar spine (5 to 21). Sagittal measurements for the changes in the arc of the upper and lower screws on the construct were 4 degrees in the thoracic group (2 to 8) and 9 degrees in the lumbar group (2 to 17). By latest follow-up 11 patients (38%) underwent reoperation, with most cases due to implant breakage (N=4, 14%), extender misalignment (N=2, 7%), and screw misplacement (N=2, 7%).

Conclusion:

At mean 1.9 years postoperatively, PDDD preserves measurable spinal motion over the construct both in the coronal and the sagittal plane without evidence for autofusion. Coronal arc of motion averages 10 to 12 degrees and sagittal arc of motion ranged from 4 to 14 degrees, although this varies by patient. This study confirms that PDDD for pediatric scoliosis preserves a measurable degree of postoperative flexibility both in the sagittal and coronal planes.

Level of Evidence:

Level II—therapeutic study.
Current trends and advances in antiviral therapy for chronic hepatitis B
Authors: Li Juan , Liu Siyi , Zang Qijuan , Yang Ruijie , Zhao Yingren , He Yingli
Published: July 2024

Chronic hepatitis B virus (HBV) infection is a global public health concern. Existing antiviral drugs, including nucleos(t)ide analogs and interferon-α, can suppress HBV replication and improve the prognosis. However, the persistence of covalently closed circular DNA (cccDNA), the integration of HBV-DNA into the host genome, and compromised immune responses impede the successful treatment of hepatitis B. While achieving a functional cure of HBV remains elusive with the current treatment methods, this is the goal of new therapeutic approaches. Therefore, developing novel antiviral drugs is necessary for achieving a functional or complete cure for chronic hepatitis B. In recent years, substantial progress has been made in drug discovery and development for HBV infection. Direct-acting antiviral agents such as entry inhibitors, capsid assembly modulators, subviral particle release inhibitors, cccDNA silencers, and RNA interference molecules have entered clinical trials. In addition, several immunomodulatory agents, including toll-like receptor agonists, therapeutic vaccines, checkpoint inhibitors, and monoclonal antibodies, are also making their way toward clinical use. In this review, we summarize the recent progress and limitations of chronic hepatitis B treatment and discuss perspectives on approaches to achieving functional cure. Although it will take some time for these new antiviral drugs to be widely used in clinical practice, combination therapy may become a preferable treatment option in the future.
Hormonal influences on meningioma risk in women
Authors: Giraldi Laura , Dalhoff Kim P.
Published: July 2024

Meningiomas have been linked to endogenous and exogenous hormones, but the exact mechanisms are unclear. A review of 23 studies (1958–2018) found no significant association between menarche age and meningioma risk. Most contraceptives showed no link, though some specific types indicated high risk. Pregnancy might be protective while breastfeeding over 6 months showed a protective effect in one study. Postmenopausal status increased risk in two studies. Hormone replacement therapy (HRT) results were inconsistent, with some studies indicating increased risk, particularly with long-term use. High cumulative doses of cyproterone acetate (CPA) were significantly associated with increased risk. Overall, hormonal factors’ role in meningioma development varies, with specific hormones and high doses of CPA posing significant risks, suggesting the need for further research.
Causal relationships between inflammatory cytokines and myopia: an analysis of genetic and observational studies
Authors: Liang Rongbin , Li Tao , Gao Hui , Shi Wenqing , Li Meilin , Wan Ting , Zhou Xiaodong
Published: July 2024

Objective:

This study aims to explore the causal relationship between inflammatory markers and myopia through the use of bidirectional Mendelian randomization (MR) and myopia animal models.

Methods:

The authors utilized data from a comprehensive and publicly accessible genome-wide association study (GWAS) for our analysis, which includes 460 536 European ancestry control subjects and 37 362 myopia patients. Utilizing a two-sample Mendelian randomization analysis framework, 27 inflammatory markers were investigated as exposure variables with myopia serving as the outcome variable. Nine MR analysis techniques were employed, with inverse-variance weighting (IVW) as the principal MR analysis method. Heterogeneity was assessed using Cochrane’s Q test. The identification of single-nucleotide polymorphisms (SNPs) and outliers linked to myopia was achieved via MR-PRESSO. The expression of interleukin-2 (IL-2) in the vitreous of guinea pigs subjected to experimentally induced form-deprivation myopia (FDM) was examined.

Results:

Elevated concentrations of IL-2 and IL-2ra were found to be associated [IVW estimate odds ratio (OR): 1.003, 95% CI: 1.001–1.005, P=0.001] and strongly associated (IVW estimate OR: 1.002, 95% CI: 1.000–1.003, P=0.049) with an increased risk of myopia, respectively. Conversely, lower levels of C-reactive protein (CRP) (IVW estimate OR: 0.996, 95% CI: 0.994–0.999, P=0.002) and tumour necrosis factor alpha (IVW estimate OR: 0.995, 95% CI: 0.994–0.996, P<0.001) were robustly linked to a heightened risk of myopia. IL-2 expression was notably upregulated in the vitreous of guinea pigs with experimentally induced FDM.

Conclusions:

Elevated levels of inflammatory factors, especially IL-2 and IL-2ra, have a potential causal relationship with myopia susceptibility, providing new insights into the pathogenesis of myopia.
Can Patients With Narcissistic Personality Disorder Change? A Case Series
Authors: Weinberg Igor , Ronningstam Elsa , Ravichandran Caitlin , Gunderson John G.
Published: July 2024

The study was set out to establish the potential for psychotherapy to effect improvements in patients with narcissistic personality disorder (NPD). Eight patients with NPD who improved in treatment were identified. Consensus clinician/investigator diagnostic scores from before and after the psychotherapies were retroactively established on the Diagnostic Interview for Narcissism (DIN) and the Diagnostic Statistic Manual for Psychiatric Disorders, 5th Edition (DSM-5) Personality Disorder Section II criteria. Psychosocial functioning (work or school, romantic relationships) before and after the psychotherapies was retroactively evaluated as well. At the completion of the therapies after 2.5 to 5 years, all patients had improved, no longer met DIN or DSM-5 criteria for NPD, and showed better psychosocial functioning. Symptomatic improvements were associated with large effect sizes. In conclusion, changes in NPD can occur in treatment after 2.5 to 5 years. Future research should identify patient characteristics, interventions, and common processes in such improved cases that could help with development of treatments.
Scoping review: Reporting characteristics for the safety of contact lenses in the pediatric population
Authors: Tomiyama Erin S. , Kobia-Acquah Emmanuel , Ansari Shora M. , Logan Anna-Kaye , Gialousakis John , Ng Sueko , Wagner Heidi
Published: July 2024

SIGNIFICANCE

Contact lenses are an increasingly popular option for correcting pediatric refractive error due to increased awareness of interventions to slow myopia progression. With limited information on the safety profiles of contact lenses in children, it is important to characterize the current understanding and promote this device's safe and effective use.

PURPOSE

This scoping review evaluates characteristics of the current literature that have examined the safety of contact lenses in pediatric patients. It provides future directions for systematic reviews and identifies any gaps in the current literature or areas for future research.

METHODS

Literature searches in MEDLINE via PubMed, EMBASE, The Cochrane Library, trial registries, and U.S. Food and Drug Administration clinical trial documentation were performed. Included studies (i.e., experimental and quasi-experimental studies; observational studies including prospective and retrospective cohort, case-control, and analytical cross-sectional studies, and case series of 30 or more participants) reported safety and/or complications of the use of any contact lens for correcting refractive error in children (0 to 18 years). Two independent reviewers first screened the titles and abstracts, and then full-text reports for eligibility. Conflicts in eligibility were resolved by discussions with a third reviewer. Two independent reviewers extracted data, including details about the participants, context, study methods, and key findings relevant to the review question.

RESULTS

This scoping review included 73 studies from 10 countries using different contact lens modalities, primarily orthokeratology and soft contact lenses, in children (6 to 18 years). The most common adverse event reported by the studies was corneal staining (60% orthokeratology, 45% soft contact lens).

CONCLUSIONS

The need for uniform reporting standards for adverse events poses challenges for comprehensive data synthesis. However, this scoping review identified a sufficient number of studies for a future systematic review to quantify the risks associated with orthokeratology and soft contact lens use in children.
Splinting of Low-Risk Pediatric Fractures
Authors: Miller Andrew F. , Hannon Megan M.
Published: July 2024

This review covers common orthopedic injuries seen after acute traumatic injury. A thorough physical examination and radiographic review of these injuries are key to ensuring appropriate management. Although many injuries may require urgent or emergent orthopedic consultation and management, this review focuses on injuries that are low-risk and amenable to splinting with outpatient orthopedic follow-up. This review covers key physical examination features, radiographic findings that help providers assess injuries, as well as reviewing splinting application to help facilitate rapid management of these injuries in the acute care setting.
Multifrequency Magnetic Resonance Elastography Detects Small Abdominal Lymph Node Metastasis by High Stiffness
Authors: Neelsen Christian , Elgeti Thomas , Meyer Tom , Grittner Ulrike , Mödl Lukas , Furth Christian , Geisel Dominik , Hamm Bernd , Sack Ingolf , Marticorena Garcia Stephan Rodrigo
Published: July 2024

Objectives

Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 is a clinical and research standard for evaluating malignant tumors and lymph node metastasis. However, quantitative analysis of nodal status is limited to measurement of short axis diameter (SAD), and metastatic lymph nodes below 10 mm in SAD are often not detected. The purpose of this study was to evaluate the value of multifrequency magnetic resonance elastography (MRE) when added to RECIST 1.1 for detection of lymph node metastasis.

Materials and Methods

Twenty-five benign and 82 metastatic lymph nodes were prospectively examined by multifrequency MRE at 1.5 T using tomoelastography postprocessing at 30, 40, 50, and 60 Hz (total scan time of 4 minutes). Shear wave speed as a surrogate of soft tissue stiffness was provided in m/s. Positron emission tomography–computed tomography was used as reference standard for identification of abdominal lymph node metastasis from histologically confirmed primary tumors. The diagnostic performance of MRE was compared with that of SAD according to RECIST 1.1 and evaluated by receiver operating characteristic curve analysis using generalized linear mixed models and binary logistic mixed models. Sensitivity, specificity, and predictive values were calculated for different cutoffs.

Results

Metastatic lymph nodes (1.90 ± 0.57 m/s) were stiffer than benign lymph nodes (0.98 ± 0.20 m/s, P < 0.001). An area under the curve of 0.95 for a cutoff of 1.32 m/s was calculated. Using a conservative approach with 1.0 specificity, we found sensitivity (SAD/MRE/MRE + SAD, 0.56/0.84/0.88), negative predictive values (0.41/0.66/0.71), and overall accuracy (0.66/0.88/0.91) to be improved using MRE and even higher for combined MRE and SAD.

Conclusions

Multifrequency MRE improves metastatic abdominal lymph node detection by 25% based on higher tissue stiffness—even for lymph nodes with an SAD ≤10 mm. Stiffness information is quick to obtain and would be a promising supplement to RECIST.
The Craniofacial Collaboration UK: Developmental Outcomes in 7- and 10-Year-Old Children With Sagittal Synostosis
Authors: Lloyd-White Samuel , Samuel Rosanna , Edwards-Bailey Laura , Piggott Katie , Blighe Sabrina , Bassi Amber , Hotton Matthew , Horton Jo , Johnson David , Parks Christopher , Kearney Anna
Published: July 2024

The Craniofacial Collaboration UK (CC-UK) protocol is a shared agreement across the 4 UK Highly Specialist Craniofacial Centres (HSCCs) to conduct robust neurodevelopmental and psychosocial clinical screening for children with craniosynostosis. This agreement allows for the analysis of outcomes of a homogenous sample of children with single suture craniosynostosis (SSC), a frequent limitation of the existing research. The current study is the latest analysis of CC-UK data on behavioral, cognitive, and psychosocial outcomes. The focus of this analysis is 7- and 10-year-olds with nonsyndromic sagittal synostosis (SS) who have undergone primary corrective surgery and completed routine clinical screening at 1 of the 4 HSCCs since the introduction of the CC-UK protocol. Due to changes in clinical pathways, only data from 3 HSCCs is included to preserve homogeneity. Results show that the majority of children with SS fall within the average range across behavioral and neurodevelopmental domains. A notable exception was a task involving perceptual reasoning and visuomotor skills (Block Design). Although this difference was small and the mean score remained within the average range, it suggests some increased risk of subtle difficulty with such skills for children with SS. Across other measures, there was no consistent evidence of any significantly increased risk of poorer outcomes, in line with findings of previous CC-UK papers. Understanding the psychological phenotype of SS is a key research priority for parents and clinicians, and the current study is another step toward achieving this goal.
Bilateral Temporomandibular Joint Reconstruction With Custom Alloplastic Implants for Pediatric Ankylosis
Authors: Alba Brandon , Harmon Kelly A. , La-Anyane Okensama , Jones Kody , Figueroa Alvaro , Tragos Christina
Published: July 2024

For pediatric patients with refractory temporomandibular joint (TMJ) ankylosis, reconstruction with autologous techniques such as costochondral grafts or distraction osteogenesis has long been considered the gold standard. Many surgeons believed the use of alloplastic joint replacement to be contraindicated in pediatric patients due to concerns for growth restriction and the limited lifespan of the implants. However, recent data has supported TMJ prostheses in skeletally immature patients. This study aims to present a case series of pediatric patients undergoing bilateral TMJ reconstruction with custom-made implants and evaluate their postoperative results. A retrospective chart review was performed of all consecutive pediatric patients undergoing bilateral alloplastic TMJ reconstruction for refractory ankylosis. All patients underwent bilateral TMJ release and total joint replacement with custom-made implants. Preoperative and postoperative cephalometric and volumetric airway data was obtained using cone-beam computed tomography. Three patients, aged 8 to 17, underwent bilateral TMJ replacement with custom-made implants. There were no postoperative complications, and no implants required explantation or replacement. Postoperatively, all patients had increases in maximal interincisal opening, which was stable over months/years of follow-up. The patients also subjectively reported improved speech and mastication; 1 patient had significant improvements in sleep apnea symptoms. Volumetric airway analysis revealed an average airway size increase of 25.6%. Alloplastic TMJ reconstruction is a safe, effective solution for refractory ankylosis in pediatric patients and represents a promising new frontier in craniofacial surgery. Continued long-term evaluation will provide further evidence of the utility of this procedure.
Systematic Reporting of Eosinophils in Transbronchial Biopsies After Lung Transplantation Defines a Distinct Inflammatory Response
Authors: Darley David R. , Sivasubramaniam Vanathi , Qiu Min R. , Barrett Wade A. , Wong Stephen J. , Martinu Tereza , Pal Prodipto , Thwe Le Myo , Tonga Katrina O. , MacDonald Peter S. , Plit Marshall L.
Published: July 2024

Background.

Descriptions of eosinophils in transbronchial biopsy (TBBx) pathology reports after lung transplantation (LTx) are associated with poor long-term outcomes. The absence of routine reporting and standardization precludes accurate assessment of this histologic predictor. A systematic reporting scheme for the presence of TBBx eosinophils after LTx was implemented. This report aims to assess this scheme by describing the presence, pattern, and gradation of TBBx eosinophils and clinical associations.

Methods.

A prospective cross-sectional study of all TBBx reports was performed including all patients presenting for a surveillance or diagnostic TBBx between January 2020 and June 2023. Each TBBx was systematically reported in a blinded manner. Mixed-effects logistic regression was performed to measure the association between concurrent clinical and histologic features, and the presence of TBBx eosinophils.

Results.

A total of 410 TBBx reports from 201 patients were systematically reported. In 43.8% recipients, any TBBx eosinophils were detected and in 17.1% recipients, higher-grade eosinophils (≥3 per high power field) were present. Adjusted analysis showed that retransplantation, A- and B-grade cellular rejection, positive bronchoalveolar lavage (BAL) bacterial microbiology, and elevated blood eosinophil count were independently associated with the presence of any TBBx eosinophils. Diagnostic “for-cause” procedures were independently associated with higher quantities of TBBx eosinophils.

Conclusions.

Systematic reporting demonstrates that TBBx eosinophils are a distinct inflammatory response associated with rejection, infection, and peripheral eosinophilia. Although these findings require multicenter external validation, standardized reporting for TBBx eosinophils may assist in identifying recipients at risk of poor outcomes and provides a platform for mechanistic research into their role after lung transplantation.
Impact of varicocelectomy on sperm parameters and hormone levels in infertile patients with sickle cell disease
Authors: Ghanem Ahmed M. , Ghanem Mazen A. , Ghanem Ahmed A.
Published: July 2024

Purpose:

This study analyzes the semen parameters and gonadal hormone levels in infertile patients with sickle cell disease (SCD) and palpable varicoceles who underwent varicocelectomy for impaired semen parameters.

Patients and methods:

Improvements in the semen parameters—serum levels of total testosterone, follicle-stimulating hormone (FSH), and inhibin B—and testicular volume (TV) in 18 infertile patients with SCD were evaluated before and at 6 months after varicocelectomy.

Results:

Before varicocelectomy, all sperm parameters were affected, with high rates of low sperm concentration (83.3%), poor motility (88.9%), and abnormal morphology (77.8%). All sperm parameters were altered in 61.1%. On follow-up at 55.5 months after varicocelectomy, improvement in all the semen parameters was successful and complete in 22.2% but was not significant in 77.8%. The median follow-up period for sperm recovery was relatively long at 60.3 months. Total testosterone, FSH, and inhibin B were not significantly different before and after treatment (P > 0.05). However, testosterone, FSH, and TV significantly correlated with the semen parameters (P < 0.05). Moreover, a significant positive correlation was found between TV and inhibin B (P < 0.05), and the improvement in the semen parameters was significantly correlated with primary hypogonadism (P < 0.05).

Conclusion:

In adult men with SCD, varicocelectomy had a positive impact on the impaired seminal parameters and may be an option before sperm cryopreservation to improve future male fertility. Furthermore, primary testicular failure may be a feature of reversible spermatogenesis recovery in these patients.
Structure-Mechanics Principles and Mechanobiology of Fibrocartilage Pericellular Matrix: A Pivotal Role of Type V Collagen
Authors: Wang Chao , Fan Mingyue , Heo Su-Jin , Adams Sheila M. , Li Thomas , Liu Yuchen , Li Qing , Loebel Claudia , Alisafaei Farid , Burdick Jason A. , Lu X. Lucas , Birk David E. , Mauck Robert L. , Han Lin
Published: July 2024

The pericellular matrix (PCM) is the immediate microniche surrounding resident cells in various tissue types, regulating matrix turnover, cell-matrix cross-talk and disease initiation. This study elucidated the structure-mechanical properties and mechanobiological functions of the PCM in fibrocartilage, a family of connective tissues that sustain complex tensile and compressive loads in vivo. Studying the murine meniscus as the model tissue, we showed that fibrocartilage PCM contains thinner, random collagen fibrillar networks that entrap proteoglycans, a structure distinct from the densely packed, highly aligned collagen fibers in the bulk extracellular matrix (ECM). In comparison to the ECM, the PCM has a lower modulus and greater isotropy, but similar relative viscoelastic properties. InCol5a1+/Dmenisci, the reduction of collagen V, a minor collagen localized in the PCM, resulted in aberrant fibril thickening with increased heterogeneity. Consequently, the PCM exhibited a reduced modulus, loss of isotropy and faster viscoelastic relaxation. This disrupted PCM contributes to perturbed mechanotransduction of resident meniscal cells, as illustrated by reduced intracellular calcium signaling, as well as upregulated biosynthesis of lysyl oxidase and tenascin C. When cultured in vitro,Col5a1+/Dmeniscal cells synthesized a weakened nascent PCM, which had inferior properties towards protecting resident cells against applied tensile stretch. These findings underscore the PCM as a distinctive microstructure that governs fibrocartilage mechanobiology, and highlight the pivotal role of collagen V in PCM function. Targeting the PCM or its molecular constituents holds promise for enhancing not only meniscus regeneration and osteoarthritis intervention, but also addressing diseases across various fibrocartilaginous tissues.
Transcranial focused ultrasound activates feedforward and feedback cortico-thalamo-cortical pathways by selectively activating excitatory neurons
Authors: Gao Huan , Ramachandran Sandhya , Yu Kai , He Bin
Published: July 2024

Transcranial focused ultrasound stimulation (tFUS) has been proven capable of altering focal neuronal activities and neural circuits non-invasively in both animals and humans. The abilities of tFUS for cell-type selection within the targeted area like somatosensory cortex have been shown to be parameter related. However, how neuronal subpopulations across neural pathways are affected, for example how tFUS affected neuronal connections between brain areas remains unclear. In this study, multi-site intracranial recordings were used to quantify the neuronal responses to tFUS stimulation at somatosensory cortex (S1), motor cortex (M1) and posterior medial thalamic nucleus (POm) of cortico-thalamo-cortical (CTC) pathway. We found that when targeting at S1 or POm, only regular spiking units (RSUs, putative excitatory neurons) responded to specific tFUS parameters (duty cycle: 6%-60% and pulse repetition frequency: 1500 and 3000 Hz ) during sonication. RSUs from the directly connected area (POm or S1) showed a synchronized response, which changed the directional correlation between RSUs from POm and S1. The tFUS induced excitation of RSUs activated the feedforward and feedback loops between cortex and thalamus, eliciting delayed neuronal responses of RSUs and delayed activities of fast spiking units (FSUs) by affecting local network. Our findings indicated that tFUS can modulate the CTC pathway through both feedforward and feedback loops, which could influence larger cortical areas including motor cortex.
Cancer vaccine attenuates carcinogen induced head and neck cancer with impaired early T cell response
Authors: Kono Michihisa , Rokugo Masahiro , Quadarella John D. , Saito Shin , Komatsuda Hiroki , Fu Cong , Woo Sook-Bin , Egloff Ann Marie , Uppaluri Ravindra
Published: July 2024

Effective T cell immunotherapy requires understanding antigen-specific T cell development during tumorigenesis and immune surveillance. Here, we aimed to examine the dynamics of antigen-specific T cells from tumor initiation through progression in a tobacco carcinogen mimetic, 4-nitroquinoline-1-oxide (4NQO)-induced head and neck carcinogenesis model utilizing genetically engineered K5CreERT/+/ROSAOVA-GFP/p53fl/fl(KOG) mice. Our findings showed that early ovalbumin (OVA) expression via direct lingual tamoxifen (T) did not impact cancer development and survival, by comparing mice with tongue epithelium expressing OVA (KOG/T/OVA+) to those without OVA (KOG/T/OVA-) controlled by doxycycline. This equivalent tumor growth cannot be attributed to the loss of OVA expression. Intriguingly, although OVA-specific T cells were initially generated in tumor-draining lymph nodes (TDLN), they became undetectable 3 weeks after tamoxifen injection. Moreover, therapeutic anti-PD-1 was unable to restore OVA-specific T cells in TDLN and did not yield anti-tumor activity. Remarkably, OVA synthetic long peptide (SLP) vaccine induced OVA-specific T cells in KOG/T/OVA+ mice, and the combination of SLP vaccine and anti-PD-1 significantly reduced tongue tumor burden and prolonged survival. This study highlights the role of impaired endogenous antigen-specific T cell responses in immune resistance in head and neck cancer and the potential of cancer vaccines to improve outcomes.
Freezing Diluted Bovine Serum Albumin Standards Does Not Significantly Affect Standard Curves
Authors: Sheets Rachelle , Rajaboina Bhavik , Bromberg Caitlin E. , Curtin Liam P. , Haddock Mitchell L. , Stafford Phillip , Thomas Theresa Currier , Scheck Adrienne C.
Published: July 2024

Total protein isolation followed by quantitation is a common protocol in many laboratories. Quantitation is often done using a colorimetric assay such as the bicinchoninic acid (BCA) assay in which a change in the color of the BCA reagent is related to protein concentration. Extracted protein samples are compared to a standard curve made with dilutions of a protein standard such as bovine serum albumin (BSA) to determine their concentrations. A series of experiments was designed to determine the most reproducible and accurate method for quantifying protein concentrations of samples in an experimental series over time. The effect of freezing on diluted standards was investigated. Standards were frozen at −20°C or −80°C and serially thawed and refrozen up to three times prior to their use in a BCA assay. Thawing and refreezing the standards had no significant effect on protein concentration and the resulting standard curves. Inter-person and intra-person variability in the preparation of standards was also investigated. Protein concentration differences due to inter-person and intra-person variability were greater than protein concentration variability resulting from freezing and thawing, regardless of the freezing temperature. The most reproducible and accurate method for determining the protein concentration of extracted samples in an experimental series over time is diluting a large batch of BSA standards and freezing them at either −20°C or −80°C. Reproducibility was maintained with up to three freeze-thaws.

Highlights

Freezing diluted BSA standards at either −20°C or −80°C and thawing and refreezing them up to three times does not significantly alter their protein concentrations.

Inter-person variability in standard curves is greater than intra-person variability, and controlling for developer decreases variability in most cases.

There is more consistency in standard curves when a single batch of diluted standards is aliquoted and frozen at either −20°C or −80°C and thawed up to three times than when the same investigator or different investigators make fresh standards before each assay.
Latent mitotic vulnerability of AML cells induced by therapeutic agents
Authors: Niikura Ryuta , Yabushita Tomohiro , Yamamoto Shohei , Suzuki Hiroaki , Fukuyama Masamitsu , Hata Shoji , Goyama Susumu , Kitamura Toshio , Chinen Takumi , Kitagawa Daiju
Published: July 2024

Acute myeloid leukemia (AML) is a hematopoietic malignancy with a poor prognosis. Understanding the unidentified properties of AML cells is beneficial for the identification of novel therapeutic strategies for AML. In this study, we uncover the vulnerabilities of AML cells in mitosis when exposed to therapeutic agents. Through comparative analysis of large-scale data quantifying drug effects on cancer cell proliferation, the drug targeting the cell cycle and mitosis are predicted to possess high cytotoxicity against AML cell lines. Consistently, live-cell imaging with microwell devices demonstrates that clinical drugs targeting the cell cycle processes, such as idarubicin, pevonedistat and vincristine, potently induce mitotic cell death in AML cells. While these therapeutic agents also induce cell death through S/G2 phase arrest, the cytotoxic effects during mitosis are notably more pronounced. Furthermore, by employing additional inhibition of Chk1 to override the G2/M checkpoint, the AML cells stalled in the S/G2 phase prematurely enter mitosis, resulting in a significant increase in cell death. Collectively, these results unveiled the latent mitotic vulnerabilities of AML cells, providing a basis for developing novel therapeutic interventions.
Flexible movement kernel estimation in habitat selection analyses with generalized additive models
Authors: Arce Guillen Rafael , Pohle Jennifer , Jeltsch Florian , Roeleke Manuel , Reineking Björn , Klappstein Natasha , Schlägel Ulrike
Published: July 2024

Habitat selection analysis includes resource selection analysis (RSA) and step selection analysis (SSA). These frameworks are used in order to understand space use of animals. Particularly, the SSA approach specifies the space availability of sequential locations through a movement kernel. This movement kernel is typically defined as the product of independent parametric distributions of step lengths (SLs) and turning angles (TAs). However, this assumption may not always be plausible for real data where short SLs are often correlated with large TAs and vice versa.

The objective of this paper is to relax the need for parametric distributions usinggeneralized additive models(GAMs) and the R-package mgcv, based on the work of Klappstein et al. (2024). For this, we propose to specify the movement kernel as a bivariate tensor product, rather than independent distributions of SLs and TAs. In addition, we account for residual spatial autocorrelation in this GAM-approach.

Using simulations, we show that the tensor product approach accurately estimates the underlying movement kernel and that the fixed effects of the model are not biased. In particular, if the data are simulated with a copula distribution for SL and TA, i.e. if the independence assumption for SL and TA does not hold, the GAM approach produces better estimates than the classical approach. In addition, including a bivariate tensor product in the model leads to a better uncertainty estimation of the model parameters and a higher predictive quality of the model.

Incorporating a bivariate tensor product solves the problem of assuming parametric distributions and independence between SLs and TAs. This offers greater flexibility and makes the analysis of real data more reliable.
Ecologically and economically sustainable level of timber harvesting in boreal forests – defining the safe operating space for forest use
Authors: Mönkkönen Mikko , Blattert Clemens , Cours Jérémy , Duflot Rémi , Elo Merja , Eyvindson Kyle , Kouki Jari , Triviño María , Burgas Daniel
Published: July 2024

Planetary-level analyses indicate that we are exceeding the ecological limits. However, we need approaches to implement global sustainability frameworks regionally, using natural resources at levels that allow for their regeneration. We present a framework to define limits beyond which ecosystems are threatened to collapse, to answer how much we can extract from ecosystems, and to manage natural resources for both human and ecosystem wellbeing. We exemplify this approach with the heath forest habitat types in Finland, which provide most of the national timber production. We use the IUCN Red List of Habitats to set favourable reference values for volume of deadwood, proportion of old-growth forest cover and proportion of deciduous trees. Through forest growth simulations and management optimization, we found that the proportion of old-growth forest is the most challenging criteria to be reached only by 2100. This would require not only a larger use of extensive forest management practices than hitherto but also to drastically reduce the maximum economic sustainable harvest level from the current 96% to 60%. By combining threat assessments with ecosystem modelling and management planning, this approach can support regional decision makers to make informed decisions to stay within safe limits of use of natural resources.
Detecting branching rate heterogeneity in multifurcating trees with applications in lineage tracing data
Authors: Gao Yingnan , Feder Alison F
Published: July 2024

Understanding cellular birth rate differences is crucial for predicting cancer progression and interpreting tumor-derived genetic data. Lineage tracing experiments enable detailed reconstruction of cellular genealogies, offering new opportunities to measure branching rate heterogeneity. However, the lineage tracing process can introduce complex tree features that complicate this effort. Here, we examine tree characteristics in lineage tracing-derived genealogies and find that editing window placement leads to multifurcations at a tree’s root or tips. We propose several ways in which existing tree topology-based metrics can be extended to test for rate heterogeneity on trees even in the presence of lineage-tracing associated distortions. Although these methods vary in power and robustness, a test based on theJ1statistic effectively detects branching rate heterogeneity in simulated lineage tracing data. Tests based on other common statistics (ŝand the Sackin index) show interior performance toJ1. We apply our validated methods to xenograft experimental data and find widespread rate heterogeneity across multiple study systems. Our results demonstrate the potential of tree topology statistics in analyzing lineage tracing data, and highlight the challenges associated with adapting phylogenetic methods to these systems.
An efficient method for immortalizing mouse embryonic fibroblasts
Authors: Srinivasan Srisathya , Ho Hsin-Yi Henry
Published: July 2024

Mouse embryonic fibroblasts (MEFs) derived from genetically modified mice are a valuable resource for studying gene function and regulation. The MEF system can also be combined with rescue studies to characterize the function of mutant genes/proteins, such as disease-causing variants. However, primary MEFs undergo senescence soon after isolation and passaging, making long-term genetic manipulations difficult. Previously described methods for MEF immortalization are often inefficient or alter the physiological properties of the cells. Here, we describe an optimized protocol for immortalizing MEFs via CRISPR-mediated deletion of theTp53gene. This method is highly efficient and consistently generates immortalized MEFs, or iMEFs, within 14 days. Importantly, iMEFs closely resemble the parent cell populations, and individual iMEFs can be cloned and expanded for subsequent genetic manipulation and characterization. We envision that this protocol can be adopted to immortalize other mouse primary cell types.

Key Features

CRISPR-based knockout of theTp53gene enables efficient immortalization of mouse embryonic fibroblasts (MEFs) in under 2 weeks.

Immortalization requires a Neon electroporator or another comparable electroporation system to transfect cells with theTp53CRISPR constructs.
Transmission is a key driver of extensively drug-resistant tuberculosis
Authors: Goig Galo A. , Loiseau Chloé , Maghradze Nino , Mchedlishvili Kakha , Avaliani Teona , Brites Daniela , Borrell Sonia , Aspindzelashvili Rusudan , Avaliani Zaza , Kipiani Maia , Tukvadze Nestani , Jugheli Levan , Gagneux Sebastien
Published: July 2024

Multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant (XDR) TB are threatening global TB control. The World Health Organization has recently endorsed new regimens for the treatment of MDR-TB that rely on the new and repurposed drugs bedaquiline, pretomanid and linezolid with or without moxifloxacin (BPaL(M)). As BPaL(M) is being rolled-out, resistance to these new drugs is already emerging, leading to acquired XDR-TB. Importantly, instances of transmitted XDR-TB have been reported. The spread of highly drug-resistantM. tuberculosis(MTB) strains pose at risk novel TB treatments that took decades to develop. In this study, we analyzed 6,926 MTB genomes from a 13-year nationwide study in Georgia, a known geographical hotspot of MDR-TB, together with more than 80,000 MTB genomes from public sources to estimate the relative contribution of transmission to the burden of XDR-TB. We show that XDR-TB is already geographically widespread, occurring in at least 27 countries across four continents. Moreover, we estimated that a quarter of the XDR-TB cases identified are likely the consequence of transmission. Our findings call for urgent improvements in the global diagnostic capacity, infection control, and surveillance of XDR-TB.
The COVID-19 pandemic in Canada’s provinces: mitigation measures and outcomes
Authors: Grootendorst Paul
Published: July 2024

The rapid spread of SARS-CoV-2 in early 2020 forced provincial health authorities across Canada to quickly institute infection control measures. It is now four years since the start of the global pandemic, and an opportune time to consider how Canada’s provinces compared in their SARS-CoV-2 containment policies and the resulting impacts on mortality and economic activity. I compare provincial exposure to SARS-CoV-2 using data on the number out-of-province arrivals into each province. I compare the key containment measures used in each province, the length of time that these measures were imposed, and the uptake of COVID-19 vaccines by province. Using Statistics Canada data, I also estimate the impact of the COVID-19 pandemic on provincial crude death rates during 2020-2023, and life expectancy and gross domestic product during 2020-2022. I find substantial provincial variation in pandemic responses and outcomes. The provinces varied in their use of the most stringent public health measures. Uptake of the primary COVID vaccinations varied from 76% to 92%; booster vaccination uptake varied even more. There was also marked provincial variation in mortality and economic outcomes. While this study does not estimate the impacts of provincial response stringency and COVID vaccine uptake on mortality and other outcomes, it does provide suggestive evidence that can be formally assessed in future research.
Knowledge and occupational practices of beauticians and barbers in the transmission of viral hepatitis: a mixed-methods study in Volta Region of Ghana
Authors: Adjei-Gyamfi Silas , Korang Felix Kwame , Asirifi Abigail , Asobuno Clotilda
Published: July 2024

Background

Although Ghana is endemic for hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, the National Policy on Viral Hepatitis stipulates that there is unreliable data, limited knowledge, and deficiency in research on viral hepatitis, especially among high-risk workers in the eastern part of the country. This study therefore assessed the knowledge level and occupational practices of beauticians and barbers in the transmission of HBV and HCV in the Volta Region of Ghana.

Methods

A cross-sectional mixed methods study was conducted in Volta Ghana from April to June 2021. While an in-depth interview was used to collect data from five environmental health officers who were selected as key informants in the qualitative stage, structured questionnaires/checklists, and direct observations were employed to collect data from 340 beauticians and barbers in the quantitative stage. During the qualitative stage, the process of coding, and mind mapping via thematic analysis was carried out. Furthermore, descriptive and inferential analyses were performed using Stata version 17.0 at a 95% significance level in the quantitative stage.

Results

Most beauticians and barbers reported poor knowledge levels about HBV and HCV (67.0%), although the awareness of this viral hepatitis was high (88.2%). While almost one-third of the observed participants practiced safe occupational activities (31.5%), 29.0%, 49.4%, and 55.3% of them observed hand hygiene, wore protective clothes/gloves, and sterilized or disinfected tools after use respectively. Beauticians and barbers with higher (tertiary) education (AOR=11.4; 95%CI=1.44–27.5; p=0.021) and those with heavy workload per day (AOR=4.34; 95%CI=1.31– 14.4; p=0.016) were more likely to report good knowledge level about HBV and HCV. Additionally, beauticians were more likely to practice safe occupational activities as compared to barbers (AOR=14.2; 95%CI=4.11–28.8; p<0.001). The key informant interviews revealed that there was little or no licensing, monitoring, and training organized for beauticians and barbers.

Conclusion

Participants showed high awareness but limited knowledge about HBV and HCV. The general safety practices among the participants were poor. Our study results suggest possible viral transmission through the activities of beauticians and barbers which could be attributed to the lack of regulatory systems and training of beauticians and barbers.
COVID-19 severity and risk of SARS-CoV-2-associated asthma exacerbation by time since booster vaccination: a longitudinal analysis of data from the COVIDENCE UK study
Authors: Vivaldi Giulia , Talaei Mohammad , Pfeffer Paul E , Shaheen Seif O , Martineau Adrian R
Published: July 2024

Background

In several countries, COVID-19 booster vaccinations are offered annually to priority groups, but many people have not been vaccinated in over a year. We aimed to assess the association between time since booster vaccination and characteristics of breakthrough infection. We also assessed whether incident COVID-19 continued to associate with asthma exacerbations in boosted individuals, and whether risk of COVID-19-associated exacerbation was affected by time since vaccination.

Methods

COVIDENCE UK is a prospective, longitudinal, population-based study of COVID-19. We included adult participants who had received ≥1 booster vaccination. Time since vaccination was binarised at 6 months or 12 months according to vaccine eligibility subgroup. We used logistic, Cox, and linear regression to obtain adjusted estimates for the association between time since vaccination and breakthrough infection severity, symptom duration, and acute changes to health-related quality of life (measured by the EQ-5D-3L Index). We then assessed the association of incident COVID-19 with asthma exacerbations using multilevel mixed models, by time since vaccination.

Results

7391 boosted participants reported a breakthrough infection. Across all eligibility subgroups, greater time since vaccination associated with increased odds of infection requiring bedrest (vsmilder symptoms), with the highest odds for adults aged 65–75 years (1.83 [95% CI 1.51–2.23] when vaccinated >6 monthsvs≤6 months prior). However, we observed little evidence of association between time since vaccination and symptom duration. Vaccination >12 months prior (vs≤12 months) was associated with a small decrease in EQ-5D-3L Index among participants younger than 65 years (-0.03 points [-0.04 to -0.01]). Among 2100 participants with asthma, incident COVID-19 associated with increased risk of asthma exacerbation, both ≤12 months after vaccination (OR 5.31 [4.36–6.48]) and later (6.06 [3.23–11.38]), with a greater difference in point estimates when specifically considering severe asthma exacerbations (6.82 [4.88–9.54] for ≤12 monthsvs10.06 [3.90–25.92] for >12 months).

Conclusion

Longer time since booster vaccination consistently associates with more severe breakthrough infections, and may potentially increase risk of severe asthma exacerbations. These findings highlight the importance of ensuring those currently eligible receive their booster vaccinations, and the need for research on further vaccinations in people with asthma no longer eligible for boosters.