bluuebunny/crossref_metadata_2025 huggingface dataset:
A Molecular Array for 10-second Diagnosis of Common Spinal Tumour Types with Picosecond Infrared Laser Mass Spectrometry
Authors: Fiorante Alexa , Woolman Michael , Munoz David G , Kiyota Taira , Ye Lan Anna , Farahmand Yasmine , Vlaminck Darah , Talbot Francis O , Das Sunit , Kellett Sorcha , Giuffrida Christine , Zadeh Gelareh , Ginsberg Howard J , Aman Ahmed , Zarrine-Afsar Arash
Published: February 2025

Background

Improving the surgical outcomes for commonly occurring spinal neoplasms of extradural and intradural extramedullary origins requires precise intraoperative diagnosis provided by highly trained neuropathologists.

Methods

Through a retrospective study of n=319 patient specimens, verified where appropriate by learning curve analysis to be sufficient for statistically significant observations, we aimed to assess the utility of 10-second picosecond infrared laser mass spectrometry (PIRL-MS) for non-subjective diagnosis of major spinal tumour types of metastatic carcinoma, schwannoma and meningiomas.

Results

The sensitivity and specificity values of spinal tumour type diagnosis (based on n=182 independent specimens) were (93±1)% and (97±2)%, respectively. This classification utilizes n=41 cellular lipids including phosphatidylcholines, sphingomyelins, phosphatidylethanolamines, and ceramides whose identities were established using high-resolution tandem mass spectrometry. Furthermore, the accuracy of diagnosis of a model that contained n=97 meningioma and n=106 schwannoma was not drastically influenced by the presence of n=54 additional intradural extramedullary spinal neoplasms of myxopapillary ependymoma, neurofibroma, paraganglioma and solitary fibrous tumour types in the differential diagnosis, confirming the generalizability and robustness of the identified molecular array in rendering correct classification even in the presence of data not seen previously by the model.

Conclusions

The identified lipids form a ‘molecular array’ for robust diagnosis of meningioma and schwannoma tumours by non-pathologists in a manner similar to genomic, transcriptomic or methylomic arrays used to diagnose brain cancer types, albeit on a much faster timescale of seconds as opposed to hours.
Viral delivery of recombinases activates heritable genetic switches in plants
Authors: Chamness James C , Cody Jon P , Cruz Anna J , Voytas Daniel F
Published: February 2025

Viral vectors provide an increasingly versatile platform for transformation-free reagent delivery to plants. RNA viral vectors can be used to induce gene silencing, overexpress proteins, or introduce gene editing reagents; however, they are often constrained by carrying capacity or restricted tropism in germline cells. Site-specific recombinases that catalyze precise genetic rearrangements are powerful tools for genome engineering that vary in size and, potentially, efficacy in plants. In this work, we show that viral vectors based on tobacco rattle virus (TRV) deliver and stably express four recombinases ranging in size from ∼0.6kb to ∼1.5kb and achieve simultaneous marker removal and reporter activation through targeted excision in transgenic Nicotiana benthamiana lines. TRV vectors with Cre, FLP, CinH, and Integrase13 efficiently mediated recombination in infected somatic tissue and led to heritable modifications at high frequency. An excision-activated Ruby reporter enabled simple and high-resolution tracing of infected cell lineages without the need for molecular genotyping. Together, our experiments broaden the scope of viral recombinase delivery and offer insights into infection dynamics that may be useful in developing future viral vectors.
Society of Critical Care Medicine Guidelines for the Treatment of Heat Stroke
Authors: Barletta Jeffrey F. , Palmieri Tina L. , Toomey Shari A. , AlShamsi Fayez , Stearns Rebecca L. , Patanwala Asad E. , Siparsky Nicole F. , Badjatia Neeraj , Schultz Brian , Breighner Crystal M. , Bruno Eric , Harrod Christopher G. , Trevilian Tanya , Braz de Carvalho Leandro , Houser James , Harahus John M. , Liu Yang , Swoboda Ryan , Ruhato Banguti Paulin , Bailey Heatherlee
Published: February 2025

RATIONALE:

Predicted increases in heat-related weather phenomena will result in increasing heat exposures and heat injuries, like heat stroke. Prompt recognition, early intervention, and evidence-based management are necessary to optimize outcomes.

OBJECTIVES:

The objective of these guidelines was to develop evidence-based recommendations for the treatment of patients with heat stroke.

DESIGN:

The Society of Critical Care Medicine convened a multidisciplinary panel of 18 international clinicians, comprising expertise in critical care, emergency medicine, neurocritical care, surgery, trauma/burn surgery, sports medicine, athletic training, military medicine, nursing, pharmacy, respiratory therapy, and one patient representative. The panel also included a guidelines methodologist specialized in developing evidence-based recommendations in alignment with the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology. Conflict-of-interest policies were strictly followed during all phases of guidelines development including panel selection and voting.

METHODS:

The panel members identified Patient, Intervention, Comparison, and Outcomes questions in two main areas: cooling modalities and medications that affect temperature. A systematic review for each question was conducted to identify the best available evidence, statistically analyze the evidence, and assess the certainty of the evidence using the GRADE methodology. The GRADE evidence-to-decision framework was used to formulate the recommendations. Good practice statements were included to provide additional clinical guidance.

RESULTS:

The panel generated two strong recommendations, five good practice statements and one “only-in-the-context of research” statement. Active cooling measures are recommended over passive cooling methods, with cold- or ice-water immersion achieving the fastest cooling rate. This method should be prioritized where available. In heat stroke patients, there is no evidence to support pharmacological interventions that affect temperature control and they should be avoided.

CONCLUSIONS:

The guidelines task force provided recommendations for the management of patients with heat stroke. These recommendations should be considered along with the patient’s clinical status and available resources.
Acetylsalicylic Acid Treatment in Patients With Sepsis and Septic Shock: A Phase 2, Placebo-Controlled, Randomized Clinical Trial
Authors: Almeida Thiago M. L. , Freitas Flávio G. R. , Figueiredo Rodrigo C. , Houly Sandra G. , Azevedo Luciano C. P. , Cavalcanti Alexandre B. , Damiani Lucas P. , Svicero Bianca S. , Souza Maria A. , Bustamante Cíntia L. S. , Ramos Fernando J. S. , Alves Rafael S. V. , Atallah Fernanda C. , Jackiu Míriam , Pacheco Eduardo S. , Schmidt Raysa C. , Serra Felipe S. C. , Tomotani Daniere Y. V. , Zampieri Fernando G. , Machado Flávia R. ,   BRICNET
Published: February 2025

OBJECTIVES:

Platelets play a critical role in the inflammatory response and coagulation. We aimed to evaluate whether the use of acetylsalicylic acid (ASA) would reduce the intensity of organ dysfunction in septic patients.

DESIGN:

Randomized, blinded, parallel-group, placebo-controlled trial.

SETTING:

Five general ICUs in Brazil.

PATIENTS:

Adults with sepsis for no longer than 48 hours who had at least one severe organ dysfunction (lactate > 4 mmol/L, platelets < 100,000/mm3, Pao
2/Fio
2 ratio < 200, or septic shock).

INTERVENTIONS:

Patients were randomized to receive 200 mg of ASA or placebo for 7 days.

MEASUREMENTS AND MAIN RESULTS:

The primary outcome was the change in the Sequential Organ Failure Assessment (SOFA) score between day 0 and day 7 or date of discharge/death. Safety outcomes were major bleeding and the number of blood transfusions within 14 days. The planned sample size was 218 with interim safety analyses after enrolling 109 and 163 patients. The study was discontinued due to higher frequency of major bleeding in the ASA group. We included 166 patients (ASA: 82 patients, placebo: 84). In the adjusted analysis, there was no difference in the SOFA change between the groups (mean placebo to ASA group difference, 0.60; 95% CI, –0.55 to 1.75; p = 0.30). There were no differences in any of the secondary outcomes. In the intervention group, there were a higher number of serious adverse events (9 [11%] vs. 1 [1.2%]; p = 0.009) and major bleeding (8 [8.5%] vs. 1 [1.2%]; p = 0.02).

CONCLUSIONS:

In this population of septic patients, ASA did not reduce the intensity of organ dysfunction. ASA increased the risk of severe bleeding compared with placebo.
Association Between Type of Immunosuppression and the Incidence, Microbiology, and Outcomes of Bacterial Ventilator-Associated Lower Respiratory Tract Infections: A Retrospective Multicenter Study
Authors: Kreitmann Louis , Bayon Constance , Martin-Loeches Ignacio , Póvoa Pedro , Salluh Jorge , Rouzé Anahita , Moreau Anne-Sophie , Duhamel Alain , Labreuche Julien , Nseir Saad ,   for the Type of iMmunosuppression and the Incidence, Microbiology, and OutcomeS of ventilator-Associated lower respiratory tract infections (MIMOSA) Study Group
Published: February 2025

Objectives:

Ventilator-associated lower respiratory tract infections (VALRTIs) are among the most common ICU-acquired infections in patients receiving invasive mechanical ventilation (IMV). Immunocompromised patients may have a lower incidence of VALRTI when compared with nonimmunocompromised patients, but the influence of the type of immunosuppression on the epidemiology of VALRTI has not been investigated. The study objectives were to assess the association of the type of immunosuppression with the incidence, microbiology, and outcomes (ICU mortality, ICU length of stay, and duration of IMV) of VALRTI related to bacterial pathogens.

Design:

Multicenter, international retrospective cohort study.

Setting:

One hundred eighteen ICUs (118) in nine countries.

Patients:

Eight hundred fifty-four immunocompromised adult patients (median age, 65 yr; 57.6% males) requiring IMV for greater than 48 hours, including 162 with hematologic malignancies.

Interventions:

None.

Measurements and Main Results:

Patients with hematologic malignancies had a lower 28-day cumulative incidence of bacterial VALRTI than patients with other types of immunosuppression (13.6% vs. 20.1%; adjusted cause-specific hazard ratio, 0.61; 95% CI, 0.37–0.97), mostly due to a lower incidence of ventilator-associated pneumonia (9.3% vs. 13.9%). The proportion of VALRTI cases related to multidrug-resistant bacteria was similar between groups. Occurrence of bacterial VALRTI was associated with an increased mortality and a longer ICU length of stay, but this effect was independent of the type of immunosuppression.

Conclusions:

Patients with hematologic malignancies had a lower 28-day cumulative incidence of bacterial VALRTI than patients with other types of immunosuppression, mainly due to a lower incidence of ventilator-associated pneumonia.
Outcome Following Surgical Treatment for Chronic Pilonidal Sinus Disease: A Systematic Review of Common Surgical Techniques
Authors: Wiinblad Ida-Marie Myron , Ulrichsen Johan , Brandstrup Birgitte
Published: February 2025

BACKGROUND:

The choice of operation for chronic pilonidal sinus disease remains controversial.

OBJECTIVE:

To compare the outcomes of common operations for chronic pilonidal disease.

DATA SOURCES:

We searched PubMed, Embase, and Cochrane Library.

STUDY SELECTION:

We included randomized trials in English or Danish language, published 2002-2024 comparing operations treating chronic pilonidal disease in adults and teenagers.

INTERVENTIONS:

We compared the outcomes of secondary healing, primary midline closure, Bascom’s-, Limberg’s- and Karydakis' flap operations.

MAIN OUTCOME MEASURES:

The primary outcome was recurrence; secondary outcomes were infection, healing time, and length-of-stay. We compared recurrence and infection rates in meta-analyses for all techniques. We assessed the risk-of-bias and the quality of all trials.

RESULTS:

Fifty trials included a total of 5762 participants. In a meta-analysis, the flap-operations had fewer recurrences than primary midline closure (OR 0.31 (95% CI, 0.19 to 0.51, p < 0.01). The trials comparing Flap-operations with secondary healing were heterogeneous and did not reach significance (OR 0.38 (95% CI, 0.13 to 1.13, p = 0.08). Recurrence was similar between Limberg’s- and Karydakis' operations. Infection rates were lower for the flap-operations compared with primary closure (OR 0.33 (95% CI, 0.23 to 0.48, p < 0.01) and with secondary healing (OR 0.48 (95% CI, 0.30 to 0.77, p < 0.01). Two trials tested Bascom's procedure against Limberg’s operation without significant differences. All trials found secondary healing to have significantly longer healing times than any other operation.

LIMITATIONS:

Most studies had a high or medium risk-of-bias, resulting in very-low to low certainty of evidence. The trials generally had small numbers, short follow-ups, and no reported primary outcomes or power calculations.

CONCLUSIONS:

Primary closure and secondary healing performed poorly compared with the flap techniques. Most trials tested Limberg’s operation; only 2 tested Bascom's operation. The literature suggests the surgeon's expertise determines the choice of flap technique.
Changes in Relationship Dynamics in Men and Women After Receiving the Diagnosis of Breast Versus Prostate Cancer: A Population-based Study
Authors: Heath Frederick , Ockerman Kyle , Wiesemann Gayle S. , Safeek Rachel , Heath Marie , Momeni Arash , Spiguel Lisa R. , Kovach Stephen , Sorice-Virk Sarah
Published: February 2025

Background:

Relationship changes after cancer are prevalent yet underexplored. This investigation aimed to assess factors influencing relationship changes between participants receiving the 2 most common gender-specific cancer diagnoses: breast and prostate.

Methods:

Anonymous surveys were administered via Amazon Mechanical Turk. Eligible participants were 18 years or older and diagnosed with breast or prostate cancer. Relationship satisfaction and mental health were assessed via Personal Health Questionnaire Depression Scale (PHQ-8), General Anxiety Disorder (GAD-7), and Self-Esteem and Relationship Questionnaire.

Results:

Of the 186 study respondents, 85.4% (159) had breast cancer and 14.5% (27) had prostate cancer. More breast cancer participants reported that their relationship worsened after cancer diagnosis (breast: 40.9%, prostate: 11.1%), whereas a majority of prostate cancer patients reported improved relationships (breast: 17%, prostate: 66.7%; P < 0.001). However, most participants reported no relationship status change (breast: 66.7%, prostate: 77.8%; P = 0.508) and remained with the same partner postdiagnosis (breast: 84.9%; prostate: 77.8%). Breast cancer participants reported higher self-esteem compared with prostate cancer participants (P = 0.019). There was no significant difference in overall Self-Esteem and Relationship Questionnaire (P = 0.140), PHQ-8 (P = 0.689), and GAD-7 (P = 0.871) scores. Average PHQ-8 and GAD-7 scores indicated moderate depression (breast: 12.62, prostate: 12.88) and moderate anxiety (breast: 10.96, prostate: 11.06).

Conclusions:

Breast cancer participants reported greater perceived changes in their relationship postdiagnosis. This study supports routine, active, and pre-emptive involvement of a mental health provider for patients with cancer to improve mental health outcomes.
The Impact of a High Sensitivity Troponin HEART Pathway-Based Clinical Decision Protocol on Observation Visits
Authors: Hughes George B. , Emeli Iyesatta , Wheatley Matthew A. , Goyal Abhinav , Bryksin Janetta , Moran Timothy P. , Keadey Matthew T. , Ross Michael A.
Published: February 2025

Background:

Use of high-sensitivity troponin (hs-cTn) might lead to an increase in hospital observation visits due to a higher number of abnormal troponin levels.

Study objective(s):

To determine the impact of incorporating hs-cTn into a chest pain clinical decision protocol (CDP) on observation visits in a large academic health system.

Methods:

This is a retrospective observational cohort study of all chest pain observation patients in 4 hospitals in an academic health system over 24 months. All hospitals used the Beckman Coulter Unicel Dxi instrument, and all shared the same emergency department (ED) chest pain protocol, which used the HEART pathway and serial troponins and directed ED dispositions to either an observation stay, ED discharge, or inpatient admission. Outcomes studied before and after the introduction of an hs-cTn protocol included daily chest pain observation census, cost, observation hours, and inpatient admit rate. Census was reported as the daily chest pain observation census and as a proportion of all observation visits. Data were retrieved from a health system data warehouse and a cost accounting program.

Results:

There were 6712 chest pain observation visits over 24 months, with 4087 visits before and 2634 visits after the hs-cTn protocol implementation. Comparison groups were similar in terms of age, gender, and type of insurance. There were 10.59 (95% CI, 10.24-10.95) daily chest pain observation visits before and 7.66 (95% CI, 7.34-7.97) visits after implementation, with a 28% (95% CI, 35%-20%) decrease in the total daily census. As a portion of all observation visits, there was a 22% drop in the proportion that were observed for chest pain. The daily number of chest pain patients requiring inpatient admission was unchanged. The daily total direct cost for chest pain observation decreased with an effective daily cost savings of $4313 USD (95% CI, $1534-$6998). The total daily number of chest pain observation bed hours also decreased by 41.5 hours (95% CI, 13.4–96.4 hours).

Conclusions:

Implementation of a hs-cTn chest pain protocol was associated with a significant decrease in the number and proportion of observation visits, a decrease in total daily cost and bed hours used, and no increase in inpatient admissions.
Clinician Views of an Opioid Prescribing Report with Peer Comparisons and Patient-Reported Outcomes
Authors: Ebert Jeffrey P. , Grenader E. Madeline , Gonzales Rachel E. , Spencer Evan A. , Schroeder Devon M. , Southwick Lauren , Shofer Frances S. , Delgado M. Kit , Agarwal Anish K.
Published: February 2025

Providing feedback to clinicians on their prescribing is a promising approach to right-sizing opioid prescriptions. The present research investigated the perceived acceptability, appropriateness, helpfulness, and areas for improvement of a monthly report providing surgical clinicians feedback on their postoperative opioid prescribing relative to guidelines, peer prescribing, and patient-reported pills taken, as well as on patient-reported ability to manage pain. Between January and May 2023, surgeons, advanced practice providers, and residents who recently received these reports as part of a health system quality improvement intervention completed a survey (n = 38) or interview (n = 8). Mean (SD) acceptability of the prescribing report was 4.2 (0.8), and appropriateness was 4.2 (0.8); appropriateness varied by clinical role. All features of the report were rated as “very” or “extremely” helpful by a majority of respondents. Interviewees wished for fuller explanations, real-time updates, and improved accuracy. These findings can inform the design of clinician feedback in learning health systems.
Enhanced Bone Pseudometastasis Due to Granulocyte Colony-stimulating Factor
Authors: Koyasu Sho , Suzuki Chiaki , Honda Keigo , Okuyama Chio , Nakamoto Yuji
Published: February 2025

A 48-year-old man with advanced oral cancer underwent FDG-PET/MRI after receiving neoadjuvant chemotherapy. Due to the heterogeneous and intense FDG uptake in multiple bones, including the ribs and clavicles, bone metastasis was initially considered. However, based on the pattern of FDG accumulation, MRI findings and his history of heavy drinking, we considered reconverted bone marrow (“bone pseudometastasis”) enhanced by granulocyte colony-stimulating factor (G-CSF). Biopsy confirmed hematopoietic bone marrow without malignancy. In this case, G-CSF stimulated bone marrow simulated bone metastases on FDG-PET, more strongly mimicked bone metastases than normally observed after G-CSF.
The Phoenix Sepsis Score in Pediatric Oncology Patients With Sepsis at PICU Admission: Test of Performance in a European Multicenter Cohort, 2018–2020
Authors: Wösten-van Asperen Roelie M. , la Roi-Teeuw Hannah M. , Tissing Wim J.E. , Jordan Iolanda , Dohna-Schwake Christian , Bottari Gabriella , Pappachan John , Crazzolara Roman , Amigoni Angela , Mizia-Malarz Agnieszka , Moscatelli Andrea , Sánchez-Martín María , Willems Jef , Schlapbach Luregn J. ,   on behalf of PICU Oncology Kids in Europe Research (POKER) group research consortium of European Society for Paediatric and Neonatal Intensive Care (ESPNIC)
Published: February 2025

Objectives:

The Pediatric Sepsis Definition Task Force developed and validated a new organ dysfunction score, the Phoenix Sepsis Score (PSS), as a predictor of mortality in children with suspected or confirmed infection. The PSS showed improved performance compared with prior scores. However, the criteria were derived in a general pediatric population, in which only 10% had cancer. Given that pediatric cancer patients with sepsis have higher mortality compared with noncancer patients with sepsis, we aimed to assess the PSS in PICU patients with cancer and sepsis.

Design:

Retrospective multicenter cohort study.

Setting:

Twelve PICUs across Europe.

Patients:

Each PICU identified patients 18 years young or younger, with underlying malignancy and suspected or proven sepsis, and admission between January 1, 2018, and January 1, 2020.

Interventions:

None.

Measurements and Main Results:

The PSS and three other scores, including Phoenix-8, Pediatric Logistic Organ Dysfunction-2 (PELOD-2) score, and pediatric Sequential Organ Failure Assessment (pSOFA) score, were calculated for comparison. The primary outcome was 90-day all-cause mortality. We compared score performance using area under the receiver operating characteristic curve (AUROC) and area under the precision-recall curve (AUPRC) analyses. Among 383 patients with proven or suspected sepsis, 90-day mortality was 19.3% (74/383). We failed to identify an association between a particular score and performance for 90-day mortality. The mean (95% CI) values for the AUROC of each score was: PSS 0.66 (0.59–0.72), Phoenix-8 0.65 (0.58–0.72), PELOD-2 0.64 (0.57–0.71), and pSOFA 0.67 (0.60–0.74) and for the AUPRC of each score: PSS 0.32 (0.23–0.42), Phoenix-8 0.32 (0.23–0.42), PELOD-2 0.32 (0.22–0.43), and pSOFA 0.36 (0.26–0.46). Similar results were obtained for PICU mortality or sepsis-related PICU mortality.

Conclusions:

Contrary to the general PICU population, our retrospective test of the PSS in a PICU oncology dataset with suspected or proved sepsis from European PICUs, 2018–2020, failed to identify improved performance in association with mortality. This unique patient population deserves development of organ dysfunction scores that reflect organ dysfunction and mortality data specifically from these patients and will require prospective validation in future studies.
Molecular Classification of Endometrial Carcinoma: Insights From a Teaching Hospital
Authors: Saharti Samah , Altaf Fadwa
Published: February 2025

Endometrial carcinoma is a heterogeneous disease with distinct molecular subtypes that have varied prognosis and therapeutic implications. Since the development of molecular signatures of malignancy is prominent, we are trying to implement this development in our cases of previously diagnosed endometrial cancer. The aim was to determine the prevalence of specific molecular alterations and correlate the genetic profile with the pathologic features and clinical characteristics. We identified 100 cases of endometrial carcinoma, which were eventually classified using immunostains for mismatch repair (MMR) and p53 proteins, in addition to Sanger analysis for POLE gene (Ex, 9, 13, 14). Our findings showed a high prevalence of nonspecific molecular profile (NSMP) in 46 cases (46%), and MMR deficiency in 30 cases (30%). The worst prognosis was observed in the p53 mutant pattern expressed tumors. No statistical difference in pathologic characteristics was observed when the molecular classification was applied. Of note, mutual molecular grouping assignment appears to be present in 5 (5%) of cases of endometrial carcinoma. This is the first study conducted in Saudi Arabia that investigated the prevalence and implications of these molecular subtypes in endometrial carcinoma. The percentage of cases in our result is similar to what had been published globally.
Sclerosis in Metastatic Uterine Tumor Resembling Ovarian Sex Cord Tumor: Diagnostic Dilemma Presented by Unusual Morphology
Authors: Lei Li , Aisagbonhi Omonigho
Published: February 2025

Uterine tumors resembling ovarian sex cord tumors (UTROSCT) are rare neoplasms that typically follow a benign course. However, metastasis occurs in rare cases and features associated with poor outcomes are only recently being described. These include: size >5 cm, at least moderate cytologic atypia, >3 mitosis per 10 high-powered fields, infiltrative borders, necrosis, GREB1 rearrangements, ESR1 rearrangements, and NCOA2/3 fusions. To our knowledge, prominent sclerosis has not been described in UTROSCT, nor has it been associated with an increased risk of metastasis. We present the case of a 51-yr-old woman with UTROSCT with corded/trabecular growth and sclerosis. The presence of sclerosis resulted in the misdiagnosis of her uterine tumor as leiomyoma and her lung metastasis as sclerosing epithelioid fibrosarcoma. The correct diagnosis of UTROSCT with lung metastasis was reached upon a morphologic comparison of the primary and metastatic tumors and the performance of a broad panel of immunohistochemical stains revealing the tumor to be CD99, CD56, ER, and inhibin positive and negative for rearrangements in 138 targeted genes, including genes commonly described as rearranged in endometrial stromal sarcomas, Ewing sarcoma and sclerosing epithelioid fibrosarcoma. The panel did not include GREB1 or ESR or NCOA3, but NCOA1/2 rearrangements were not detected. Our case highlights the diagnostic dilemma introduced by the presence of sclerosis in UTROSCT. We suspect prominent sclerosis may be another feature predictive of malignant potential in UTROSCT.
Radio-Guided Monitoring of Systemic Leakage With 99mTc-MAA Uptake in a Patient With Melanoma During Isolated Limb Perfusion
Authors: Samadi Mohammad Hadi , Hadad Ehsan Hasanzadeh , Khezri Susan , Sadeghi Ramin , Aghaee Atena
Published: February 2025

We present a case of a 65-year-old woman with a history of melanoma of the left foot who underwent tumor resection with inguinal lymph nodes dissection and received external beam radiation therapy. After some time, she developed nodular lesions in the same region that were confirmed to be due to involvement of the dermis. She was selected as a candidate for isolated limb perfusion procedure. The isolated limb perfusion was performed with the aid of radioguided monitoring of systemic leakage by injecting 99mTc-labeled macroaggregated albumin, and whole-body acquisition was also performed, which confirmed compartmental localization of the tracer. Our case is the first one in which 99mTc-labeled macroaggregated albumin injection has been used.
Implementation and Validation of Decision Aid Tool to Reduce Unnecessary Testing Before Thoracic Surgery
Authors: Odeh Ayham M , Verm Raymond , Kwak Jenny , Baker Marshall S , Lozanoski Madison , Helenowski Irene , Vigneswaran Wickii , Lubawski James , Raad Wissam , Freeman Richard K , Abdelsattar Zaid M
Published: February 2025

Background:

To implement and prospectively validate a previously published decision-aid tool to guide ordering of preoperative type and screen tests (preT&S).

Study Design:

In this interrupted time-series quasi-experimental study, we implemented a decision-aid tool for patients undergoing elective thoracic surgery at a single academic institution. Data was collected 6 months before and prospectively after implementation. The tool, a previously published nomogram, predicts the need for a preT&S using age, body mass index (BMI), planned operation, approach, and preoperative hemoglobin. We excluded patients who had prior transfusions, neoadjuvant therapy, redo operations and/or inpatient consults. We validated the tool using multivariable logistic regression, regression discontinuity, c-index, sensitivity, predictive values, and cost savings.

Results:

177 consecutive patients met inclusion criteria. 88 were after implementation, and 89 patients before. There were no differences between the groups in terms of age, sex, BMI, comorbidities, approach, nor preoperative hemoglobin (all p>0.05). Overall transfusion rates were similar (6.8% vs 6.7%; p>0.99); however, the rate of ordering preT&S was reduced significantly (94.4% vs 60.2%; p<0.001). At our institution, the decision aid tool resulted in cost savings of $25,048 over 6 months alone. With a c-index of 0.977, our validation demonstrated 100% sensitivity, 90.3% specificity, and 100% negative predictive value.

Conclusions:

Implementation and validation of the preT&S nomogram proved feasible, accurate, and resulted in reducing unnecessary testing and costs before elective non-cardiac thoracic surgery. Wider implementation has the potential for substantial cost savings.
A comparative in silico analysis of the vlhA gene regions of Mycoplasma gallisepticum and Mycoplasma synoviae isolates from commercial hen farms in Mexico
Authors: Maya-Rodríguez Linda M. , Gómez-Verduzco Gabriela , Trigo-Tavera Francisco J. , Moreno-Fierros Leticia , Rojas-Trejo Verónica , Miranda-Morales Rosa Elena
Published: February 2025

Avian mycoplasmosis, caused by Mycoplasma synoviae and Mycoplasma gallisepticum, poses significant economic challenges due to respiratory issues, reduced egg production and soft eggshells. The variable lipoprotein haemagglutinin (VlhA) protein, crucial for pathogenicity, comprises conserved (MSPB) and variable (MSPA) regions. The aim of this study was to identify the conserved region of vlhA gene sequences in field strain. We examined vlhA sequences from field strains collected in central Mexico (Jalisco and Mexico City). Specifically, we analysed 124 deformed eggs and 10 laying hens from 9 farms with Hy-line and Bovans breeds. Using PCR targeting the mgc2 and 16S rRNA genes, we characterized 24 field strains, 4 of which were Myc. synoviae and 20 of which were Myc. gallisepticum. We analysed the vlhA regions, based on the AF035624.1 reference sequence, with American Type Culture Collection strains as positive controls. Additionally, we validated the PCR with 20 negative samples from Mycoplasma isolation without the need for cultivation. We identified two amplification regions: MSPB and MSPA. Bioanalysis revealed relationships between our field samples and avian Mycoplasma sequences in GenBank, alongside similarities with lipoproteins present in Acholeplasma laidlawii PG8 and Escherichia coli. Given the significance of the VlhA protein in pathogenicity and immune evasion, the identified conserved sequences hold potential as therapeutic targets and for phylogenetic studies.
Rationally Designed PKD1 Activator Protects Against Neurodegeneration in Pre-clinical Models of Parkinson’s Disease
Authors: Asaithambi Arunkumar , Jang Ahyoung , Ghosh Anamitra , Ay Muhammet , Jin Huajun , Anantharam Vellareddy , Kanthasamy Arthi , Kanthasamy Anumantha G.
Published: February 2025

Oxidative stress leads to degeneration in Parkinson’s disease (PD). The key signal transduction and regulatory networks that are involved during this degenerative process in PD are currently being investigated for novel neuro-protective strategies. We recently discovered that the activation of Protein Kinase D1 (PKD1) acts as a novel compensatory mechanism in PD models and positive modulation of PKD1 can be a therapeutic strategy. Therefore, the purpose of the present study was to take a translational approach by developing a PKD1 activator and characterizing the protective function in pre-clinical models of PD. Positive genetic modulation of PKD1 by overexpression of constitutively active PKD1 protected against MPP+induced dopaminergic neurotoxicity. Pharmacological activation by Rosiglitazone protected, whereas inhibition by kb NB 142-70 exacerbated against MPP+and 6-OHDA toxicity in cell culture PD models. Importantly, peptides were rationally designed and screened for their ability to activate PKD1 using our screening methods. Peptide AK-P4 was identified to activate PKD1 specifically and protect against MPP+and 6-OHDA in both N27 cells and primary mesencephalic neurons. Further AK-P4 tagged with TAT sequence (AK-P4T) delivered using intra-venous injections activated PKD1 in mice. The neuro-protective effects of AK-P4T were tested using the sub-chronic MPTP mice model. Co-treatment with AK-P4T significantly restored the neurotransmitter levels and the behavioral and locomotory activities of the MPTP mouse model of PD. Collectively, our results demonstrate that rationally designed PKD1 activator peptide AK-P4T positively modulated PKD1 and protected against neurodegeneration in the pre-clinical models of PD. Our results suggest that positive modulation of the PKD1 using AK-P4T shows promise as a potential therapeutic agent against PD.
N-Glycan Fingerprinting of the NIST monoclonal antibody (NISTmAb)
Authors: Wu Zhengliang L
Published: February 2025

Glycosylation on therapeutic antibodies is critically important for their drug efficacies. Here, using NISTmAb and Humira® as examples, we present methods of glycan fingerprinting for these antibodies. Glycans are first released with PNGase F or Endo S2, and then labeled by specific glycosyltransferases, including sialyltransferase ST6Gal1, fucosyltransferase FUT9, N-acetyl-glucosaminyltransferase MGAT3 and fucosyltransferase FUT8, with respective fluorophore-conjugated donor sugars. The labeled glycans are then separated by gel electrophoresis (SDS-PAGE). A fluorophore-labeled hyaluronan ladder is run along with the samples to reveal the relative mobility of each glycan band. Pretreatment of the samples with specific glycosidase or glycosyltransferase results in additional mobility shift of specific glycan bands, which allows identification of some of these bands. Particularly, we report the identification of α-Gal epitopes, core-6 fucosylated glycans, paucimannose glycans and likely some bisecting/tri-antennary glycans on NISTmAb. Overall, our methods could serve as quick assessments of glycosylation on therapeutic antibodies.
Increased TGF β /Activin-Smad2 signaling is associated with pancreatic β -cell dysfunction and glucose intolerance in gestational diabetes mellitus
Authors: Boronat-Belda Talía , Ferrero Hilda , Soriano Sergi , Ribes-García Elena , Betoret-Gustems Rubén , Martínez-Bañón Daniel , Serrano-Selva Mónica , Martínez-Pinna Juan , Nadal Ángel , Quesada Iván , Magdalena Paloma Alonso-
Published: February 2025

Background

Gestational diabetes mellitus (GDM) is the most common metabolic disease during pregnancy and increases the prevalence of type 2 diabetes in both mothers and offspring. GDM management provides a window of opportunity to prevent and lower the global burden of diabetes across life. Molecular mechanisms underlying GDM are poorly defined. In this study, we explore the potential involvement of transforming growth factor beta (TGF-β) signaling in GDM as this pathway has been reported to affect pancreatic β-cell development, proliferation and identity.

Methods

We developed a GDM animal model. Serum circulating levels of TGFβ family ligands were measured in mice and human GDM. Pancreatic TGFβ signaling was investigated at the level of gene and protein expression.

Results

Our GDM animal model recapitulates the main pathophysiological features of human GDM including glucose intolerance, decreased insulin sensitivity and pancreatic β-cell malfunction. Islets from GDM mice showed impaired insulin secretion and content, altered ion channel activity, and decreased β-cell replication rate. This was accompanied by increased Smad2 signaling activation. Elevated serum activin-A and inhibin levels were found in mice and human GDM, suggesting their role as upstream signaling transducers of pancreatic Smad2 activation. Pharmacological inhibition of TGFβ/Activin-Smad2 signaling in mouse pancreatic islets resulted in improved pancreatic β-cell function and regeneration capacity of β-cells.

Conclusions

Our data disclose that disruption of pancreatic Smad2 pathway plays a critical role in the pathogenesis of GDM, contributing to abnormal glucose homeostasis and inadequate insulin secretion. Attenuation of this signaling pathway could represent a putative therapeutic target for GDM.

Highlights

High fat diet just before and during pregnancy leads to gestational diabetes in mice.

Activin and inhibin serum levels are increased in human and mice gestational diabetes.

Enhanced pancreatic Smad2 signaling contributes to inadequate insulin secretion in gestational diabetes.

Inhibition of Smad2 signaling improves pancreatic β-cell function and proliferation.
Rare missense variants of the leukocyte common antigen related receptor (LAR) display reduced activity in transcellular adhesion and synapse formation
Authors: Kaas Mathias , Chofflet Nicolas , Bicer Deniz , Skeldal Sune , Duan Jinjie , Feller Benjamin , Vilstrup Joachim , Groth Rosa , Sivagurunathan Suganya , Dashti Hesam , Pedersen Jan Skov , Werge Thomas , Børglum Anders D. , Cimini Beth A. , Jones Thouis R. , Claussnitzer Melina , Madsen Peder , Takahashi Hideto , Demontis Ditte , Thirup Søren , Glerup Simon
Published: February 2025

The leukocyte common antigen related receptor (LAR) is a member of the LAR receptor protein tyrosine phosphatase (RPTP) family of synaptic adhesion molecules that contribute to the proper alignment and specialization of synaptic connections in the mammalian brain. LAR-RPTP members have been genetically associated with neuropsychiatric disorders, but the molecular consequences of genetic perturbations of LAR remain unstudied. Using exome sequencing data from psychiatric patients and controls, we identify rare missense variants of LAR that render the extracellular domain (ECD) unstable and susceptible to proteolytic cleavage. Using recombinant and cellular systems, we describe three variants that cause disruption of the LAR:NGL-3 interaction, which results in loss of transcellular adhesion and synaptogenic effects. Furthermore, we show that overexpression of two of these variants elicit altered morphological phenotypes in an imaging-based morphological profiling assay compared to wild type LAR, suggesting that destabilization of the LAR ECD has broad effects on LAR function. In conclusion, our study identifies three rare, missense variants in LAR that could provide insights into LAR involvement with psychiatric pathobiology.
Molecular Mechanism of pH Sensing and Activation in GPR4 Reveals Proton-Mediated GPCR Signaling
Authors: You Chongzhao , Guo Shimeng , Zhang Tianwei , He Xinheng , Gao Tianyu , Xin Wenwen , Zhu Zining , Lu Yujie , Xu Youwei , Li Zhen , Zhang Yumu , Cheng Xi , Jiang Yi , Xie Xin , Xu H. Eric
Published: February 2025

Maintaining pH homeostasis is critical for cellular function across all living organisms. Proton-sensing G protein-coupled receptors (GPCRs), particularly GPR4, play a pivotal role in cellular responses to pH changes, yet the molecular mechanisms underlying their proton sensing and activation remain incompletely understood. Here we present high-resolution cryo-electron microscopy structures of GPR4 in complex with G proteins under physiological and acidic pH conditions. Our structures expose an intricate proton-sensing mechanism driven by a sophisticated histidine network in the receptor’s extracellular domain. Upon protonation of key histidines under acidic conditions, a remarkable conformational cascade is initiated, propagating from the extracellular region to the intracellular G protein-coupling interface. This dynamic process involves precise transmembrane helix rearrangements and conformational shifts of conserved motifs, mediated by strategically positioned water molecules. Notably, we discovered a bound bioactive lipid, lysophosphatidylcholine, which has positive allosteric effects on GPR4 activation. These findings provide a comprehensive framework for understanding proton sensing in GPCRs and the interplay between pH sensing and lipid regulation, offering insights into cellular pH homeostasis and potential therapies for pH-related disorders.
The T Cell-Specific miRNA–Target Network in Psoriasis: A Systematic Review
Authors: Madaan Priyanka , Tyagi Nipanshi , Tyagi Sourabh , Kushwaha Hemant Ritturaj , Jain Manju
Published: February 2025

Psoriasis is a chronic skin disease characterized by keratinocyte hyperproliferation, involving T cells as the major players in the manifestation of lesional and systemic inflammation. With the extensive exploration of keratinocyte dysfunctionality, T cell-centric deregulation has gained much focus in the context of disease etiology. miRNAs, the molecular switches known to drive disease manifestation via altering keratinocyte biology, have not been much studied in the context of T cell alterations, with only a few reports. This study aims to explore T cell-associated miRNAs and their target signaling networks to understand their role in modulating psoriasis-specific T cell changes. miRNAs with altered expression in psoriatic T cells were identified through a meta-analysis of the existing literature and datasets. These miRNAs were then used to identify cognate targets and T cell signaling pathways to elucidate T cell-specific cascades influenced by miRNAs, potentially explaining miRNA driven T cell dysregulation in psoriasis. The study identified 14 miRNAs with altered expression in psoriatic T cells and 256 downstream targets involved in 15 T cell-associated signaling pathways. The miRNA-driven targets and pathways identified may dysregulate T cell activation, proliferation, survival, and subset differentiation. These findings suggest multiple miRNA–target signaling axes that require experimental validation to better understand T cell-mediated disease etiology.
Effects of Drought on Inflorescence Yield, and Secondary Metabolites inCannabis sativaL.
Authors: Shenhar Itamar , Ifrach Idan , Barkan Ohad , Guberman Ohad , Kerem Zohar , Taler Dvir , Meiri David , Moshelion Menachem
Published: February 2025

The use of medical products derived fromCannabis sativaL. has increased significantly in recent years. While drought is known to negatively affect the yields of many crops, growers often recommend controlled periods of drought for cannabis cultivation to increase concentrations of secondary metabolites. This is especially pertinent when considering the relationship between medicinal effects and the secondary-metabolite profile.

We examined the effects of tightly controlled drought treatments on biochemical (111 phytocannabinoids and 132 terpenoids), physiological, and anatomical responses of three Type-I chemotype cultivars, specifically the THCA-dominant cultivars ‘Odem’, ‘MVA’, and ‘187’.

Our results revealed strong correlations between inflorescence and phytocannabinoid yields, on the one hand, and cumulative transpiration on the other (0.96 <r2< 0.99). Drought treatment reduced canopy conductance, with inflorescence weight decreasing by 40% and total fresh weight decreasing by 48%. The concentrations of the major phytocannabinoids, THCA and CBGA, decreased over increasing levels of drought stress (by 26% and 61%, respectively). Interestingly, terpene concentrations showed greater stress-induced variation, and that variation was genotype-dependent.

Our findings suggest that the decreases in inflorescence weight and concentrations of major phytocannabinoids under drought conditions are mainly due to a lack of biochemical-production processes, as opposed to metabolic degradation.
Cytb6/fComplex Fine-Tunes PSI Stability and Photosynthetic Capacity Under Fluctuating Light
Authors: Kono Masaru , Kodama Hiromasa , Tanigawa Keiichiro , Terashima Ichiro , Yamori Wataru
Published: February 2025

Plants encounter dynamic light fluctuations in natural habitats, requiring robust mechanisms to protect Photosystem I (PSI) against photoinhibition. The cytochrome (Cyt)b6/fcomplex plays a pivotal role in regulating electron flow and ensuring PSI stability under fluctuating light. However, its precise role in balancing PSI protection and photosynthetic capacity remains unclear. In this study, transgenic tobacco (Nicotiana tabacumL. ‘W38’) with reduced Cytb6/fcontent was used to examine PSI tolerance to photoinhibition and photosynthetic performance under fluctuating light. Notably, the reduced Cytb6/fcontent conferred significant protection of PSI against photoinhibition induced by fluctuating light, albeit at the expense of the electron transport capacity. Remarkably, PSI stability was maintained even under extreme fluctuating light conditions in the plants with lowest Cytb6/fcontents, highlighting a trade-off between photoprotection and photosynthetic capacity. These findings reveal that a reduction in Cytb6/fcontent enhances PSI protection while adjusting electron transport capacity, making plants more adaptable in environments dominated by low light and transient sunflecks. This study provides new insights into the adaptive strategies of plants under dynamic light environments and underscores the potential of targeting Cytb6/ffor enhancing crop tolerance to fluctuating light conditions.
Iterative Multiscale Molecular Dynamics: Accelerating Conformational Sampling of Biomolecular Systems by Iterating All-Atom and Coarse-Grained Molecular Dynamics Simulations
Authors: Do Hung N. , Gnanakaran S.
Published: February 2025

All-atom molecular dynamics (AA MD) simulations remain the gold standard for simulating biomolecular dynamics at an atomistic level for decades. However, they often cannot attain the time scales required to sufficiently sample the many biological processes of interest, especially those that involve large biomolecular systems with high energy barriers. Here, we combine the strengths of all-atom (AA) and coarse-grained (CG) MD simulations to develop the iterative multiscale MD (iMMD) simulation workflow in OpenMM by iterating between AA and CG MD simulations to enhance the sampling of biomolecular conformations. The AA-CG-AA iterations are repeated over multiple cycles, facilitating the accelerated sampling of biomolecular conformations at a CG level, while the finer atomistic interactions are refined with AA simulators. We demonstrate the enhanced sampling ability of iMMD over multiple AA-CG-AA iterations on four representative systems, including two globular proteins (fast-folding variant of Trpcage and Z-matrix protein of Mammarenavirus lassaense (LASV)) and two membrane proteins (Torpedo nicotinic acetylcholine receptor (nAChR) and REGN7663 Fab binding to the CXCR4 receptors in a heterogeneous cholesterol/phosphatidylcholine membrane lipid bilayer). In particular, iMMD captures the folding of the fast-folding Trpcage and Z-protein of LASV starting from their extended conformations as well as the binding of the REGN7663 Fab binding to the CXCR4s and dimerization of the CXCR4 receptors within the heterogenous membrane, within few AA-CG-AA iterations. The development of iMMD should allow for unprecedented simulations of biomolecules at much longer biological timescales to explore the biological processes of interest.