Identification of potent biparatopic antibodies targeting FGFR2 fusion driven cholangiocarcinoma.

Translocations involving FGFR2 gene fusions are common in cholangiocarcinoma and predict response to FGFR kinase inhibitors. However, the rate and durability of response are limited due to the emergence of resistance, typically involving acquired FGFR2 kinase domain mutations, and to sub-optimal dosing, relating to drug adverse effects. Here, we report the development of biparatopic antibodies targeting the FGFR2 extracellular domain (ECD), as candidate therapeutics. Biparatopic antibodies can overcome drawbacks of standard bivalent monoparatopic antibodies, which often show poor inhibitory or even agonist activity against oncogenic receptors. We show that oncogenic transformation by FGFR2 fusions requires an intact ECD. Moreover, by systematically generating biparatopic antibodies that target distinct epitope pairs along the FGFR2 ECD, we identified antibodies that effectively block signaling and malignant growth driven by FGFR2-fusions. Importantly, these antibodies demonstrate efficacy in vivo, synergy with FGFR inhibitors, and activity against FGFR2 fusions harboring kinase domain mutations. Thus, biparatopic antibodies may serve as new treatment options for patients with FGFR2-altered cholangiocarcinoma. Summary: We identify biparatopic FGFR2 antibodies that are effective against FGFR2 fusion driven cholangiocarcinoma.

Detailed Comparison of Acoustic Signals from Rehabilitated and Wild Franciscanas (Pontoporia blainvillei) Dolphins.

The franciscana dolphin is a small, vulnerable species often caught in artisanal gillnets. This study aims to provide a comprehensive assessment of their acoustic capabilities by using advanced equipment to collect a large dataset of wideband, continuous recordings. We examined the detailed acoustic signals of franciscana dolphins, comparing the sounds from rehabilitated dolphins in captivity with those of wild dolphins near fishing nets. Significant differences in acoustic characteristics were found between neonates and older dolphins, with juvenile and wild dolphins showing similar features. For the first time, repetition patterns in click production were identified, highlighting the importance of understanding the context of these sounds in regards to feeding and communication. This study emphasizes the need for detecting neonates for species protection and suggests the potential for developing acoustic classifiers specific to different age groups. Our findings offer valuable insights for conservation efforts and the development of protection strategies for franciscana dolphins.

Reduction of particle size of field peas (Pisum sativum L.) increases net energy and digestibility of starch when fed to growing pigs, but there is no difference in nutritional value between peas from the U.S. and peas from Canada.

Two experiments were conducted to test the hypothesis that particle size of field peas and location where peas are grown do not affect apparent total tract digestibility (ATTD) of nutrients and gross energy (GE), digestible energy (DE), metabolizable energy (ME), and net energy (NE), apparent ileal digestibility (AID) of starch, or standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AA). In both experiments, three sources of field peas were used. One source was obtained from the U.S., and two sources were obtained from Canada (i.e., Canada 1, Canada 2). The U.S. field peas were ground to 678, 457, or 265 µm, whereas the two sources of Canadian peas were ground to 411 and 415 µm. Therefore, five batches of field peas were used in both experiments. A basal diet contained corn and soybean meal as the only source of energy, starch, and AA, and five diets containing corn and soybean meal and 50% of each source of field peas were also formulated. The ratio between corn and soybean meal was 1.92:1 in all diets. In Exp. 1, an N-free diet was also used to calculate basal endogenous losses of AA and CP, but in Exp. 2, no N-free diet was used. In Exp. 1, seven barrows (initial body weight = 60.6 ± 2.1 kg) that had a T-cannula installed in the distal ileum were allotted to a 7 × 7 Latin square design with seven diets and seven periods. In Exp. 2, twenty-four pigs (initial body weight = 30.8 ± 1.0 kg) were housed in six calorimeter chambers with four pigs per chamber. The six chambers were allotted to one of the six diets using a 6 × 6 Latin square design with six consecutive periods of 15 d. Results of Exp. 1 demonstrated that the SID of CP and AA was not influenced by the origin of the peas or by the particle size, but the AID of starch increased (linear, P < 0.001) as particle size was reduced from 678 µm to 457 or 265 µm. Results of Exp. 2 indicated that growing location did not affect concentrations of DE, ME, or NE of field peas, but concentrations of DE, ME, and NE increased (linear, P < 0.05) when the particle size was reduced from 678 µm to 457 or 265 µm. In conclusion, field peas grown in Canada or the U.S. have the same nutritional value, but starch digestibility and NE increase if the particle size of field peas is reduced.

Allogeneic B cell immunomodulatory therapy in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is an orphan neurodegenerative disease. Immune system dysregulation plays an essential role in ALS onset and progression. Our preclinical studies have shown that the administration of exogenous allogeneic B cells improves outcomes in murine models of skin and brain injury through a process termed pligodraxis, in which B cells adopt an immunoregulatory and neuroprotective phenotype in an injured environment. Here, we investigated the effects of B-cell therapy in the SOD1G93A mouse preclinical model of ALS and in a person living with ALS. Purified splenic mature naïve B cells from haploidentical donor mice were administered intravenously in SOD1G93A mice for a total of 10 weekly doses. For the clinical study in a person with advanced ALS, IgA gammopathy of unclear significance, and B lymphopenia, CD19+ B cells were positively selected from a healthy haploidentical donor and infused intravenously twice, at a 60-day interval. Repeated intravenous B-cell administration was safe and significantly delayed disease onset, extended survival, reduced cellular apoptosis, and decreased astrogliosis in SOD1G93A mice. Repeated B-cell infusion in a person with ALS was safe and did not appear to generate a clinically evident inflammatory response. An improvement of 5 points on the ALSFRS-R scale was observed after the first infusion. Levels of inflammatory markers showed persistent reduction post-infusion. This represents a first demonstration of the efficacy of haploidentical B-cell infusion in the SOD1G93A mouse and the safety and feasibility of using purified haploidentical B lymphocytes as a cell-based therapeutic strategy for a person with ALS.

Alternative and efficient one-pot three-component synthesis of substituted 2-aryl-4-styrylquinazolines/4-styrylquinazolines from synthetically available 1-(2-aminophenyl)-3-arylprop-2-en-1-ones: characterization and evaluation of their antiproliferative activities.

In this study, an alternative and efficient one-pot three-component synthesis approach to develop a new series of (E)-2-aryl-4-styrylquinazolines and (E)-4-styrylquinazolines is described. According to this approach, the target compounds were synthesized straightforward in high yields and in short reaction times from substituted 1-(2-aminophenyl)-3-arylprop-2-en-1-ones via its well-Cu(OAc)2-mediated cyclocondensation reactions with aromatic aldehydes or its well-catalyst-free cyclocondensation reactions with trimethoxy methane (trimethyl orthoformate), and ammonium acetate under aerobic conditions. This is an operationally simple, valuable, and direct method to synthesize 2-aryl- and non-C2-substituted quinazolines containing a styryl framework at C4 position from cheap and synthetically available starting materials. All the synthesized compounds were submitted to the US National Cancer Institute for in vitro screening. The bromo- and chloro-substituted quinazolines 5c and 5d displayed a potent antitumor activity against all the tested subpanel tumor cell lines with IC50 (MG-MID) values of 5.25 and 5.50 µM, and a low cytotoxic effect with LC50 (MG-MID) values of 91.20 and 84.67 µM, respectively, indicating a low toxicity of these compounds to normal human cell lines, as required for potential antitumor agents.

Engineered Lipids for Intracellular Reactive Oxygen Species Scavenging in Steatotic Hepatocytes.

Intracellular reactive oxygen species (ROS) in steatotic cells pose a problem due to their potential to cause oxidative stress and cellular damage. Delivering engineered phospholipids to intracellular lipid droplets in steatotic hepatic cells, using the cell's inherent intracellular lipid transport mechanisms are investigated. Initially, it is shown that tail-labeled fluorescent lipids assembled into liposomes are able to be transported to intracellular lipid droplets in steatotic HepG2 cells and HHL-5 cells. Further, an antioxidant, an EUK salen-manganese derivative, which has superoxide dismutase-like and catalase-like activity, is covalently conjugated to the tail of a phospholipid and formulated as liposomes for administration. Steatotic HepG2 cells and HHL-5 cells incubated with these antioxidant liposomes have lower intracellular ROS levels compared to untreated controls and non-covalently formulated antioxidants. This first proof-of-concept study illustrates an alternative strategy to equip native organelles in mammalian cells with engineered enzyme activity.

The interaction between RIPK1 and FADD controls perinatal lethality and inflammation.

Perturbation of the apoptosis and necroptosis pathways critically influences embryogenesis. Receptor-associated protein kinase-1 (RIPK1) interacts with Fas-associated via death domain (FADD)-caspase-8-cellular Flice-like inhibitory protein long (cFLIPL) to regulate both extrinsic apoptosis and necroptosis. Here, we describe Ripk1-mutant animals (Ripk1R588E [RE]) in which the interaction between FADD and RIPK1 is disrupted, leading to embryonic lethality. This lethality is not prevented by further removal of the kinase activity of Ripk1 (Ripk1R588E K45A [REKA]). Both Ripk1RE and Ripk1REKA animals survive to adulthood upon ablation of Ripk3. While embryonic lethality of Ripk1RE mice is prevented by ablation of the necroptosis effector mixed lineage kinase-like (MLKL), animals succumb to inflammation after birth. In contrast, Mlkl ablation does not prevent the death of Ripk1REKA embryos, but animals reach adulthood when both MLKL and caspase-8 are removed. Ablation of the nucleic acid sensor Zbp1 largely prevents lethality in both Ripk1RE and Ripk1REKA embryos. Thus, the RIPK1-FADD interaction prevents Z-DNA binding protein-1 (ZBP1)-induced, RIPK3-caspase-8-mediated embryonic lethality, affected by the kinase activity of RIPK1.

Usefulness of a virtual reality educational program for reducing preoperative anxiety in children: A randomised, single-centre clinical trial.

BACKGROUND: Preoperative anxiety (PA) is common in children and has detrimental effects on surgical outcome. Strategies based on virtual reality (VR) have recently been introduced to address this problem. OBJECTIVE: This study evaluated the usefulness of a virtual reality educational program (VREP) for reducing preoperative anxiety in elective low-complexity paediatric surgery. DESIGN: Randomised clinical trial. SETTING: Single tertiary centre in Barcelona, Spain. Between January 2019 and June 2022. PATIENTS: Children aged 3-13 years of age, American Society of Anesthesiologists (ASA) I-II, scheduled for elective low-complexity surgery were enrolled in the study. INTERVENTION: Children were randomised into a control group (received oral/written information about the anaesthetic-surgical process, and patients and their parents remained in a playroom waiting for the surgery) or VREP (viewed a VR-based educational video on the surgical process, 7-10 days prior to surgery) using the MATLAB application. MAIN OUTCOME MEASURE: PA using the modified Yale Preoperative Anxiety Scale (mYPAS) during separation from parents. RESULTS: In total, 241 children aged 3-12 years of age were studied (120 patients with VREP and 121 controls). Randomisation eliminated the differences between the groups, except for a greater male presence in the VREP group (83.3% vs. 71.1%; P  = 0.023). The mYPAS yielded was lower in the VREP group (29.2% vs. 83.5%; P  < 0.001). Sex did not influence VREP-mediated decrease in PA ( P  < 0.001). In turn, VREP patients were more cooperative (Induction Compliance Checklist [ICC] score 0 points vs. 2 points; P  < 0.001) during anaesthesia induction, presented less delirium (Pediatric Anesthesia Emergence Delirium [PAED] score 1 point vs. 3 points; P  = 0.001) on leaving the recovery room, and experienced less pain upon arrival in the hospital ward (Wong-Baker Faces Pain Rating Scale: 0-points vs. 1 point; P  < 0.001). CONCLUSIONS: The VREP-based prevention strategy reduced preoperative anxiety in children undergoing elective low-complexity surgery. TRIAL REGISTRATION: NCT03578393.

Digital descriptors sharpen classical descriptors, for improving genebank accession management: A case study on Arachis spp. and Phaseolus spp.

High-throughput phenotyping brings new opportunities for detailed genebank accessions characterization based on image-processing techniques and data analysis using machine learning algorithms. Our work proposes to improve the characterization processes of bean and peanut accessions in the CIAT genebank through the identification of phenomic descriptors comparable to classical descriptors including methodology integration into the genebank workflow. To cope with these goals morphometrics and colorimetry traits of 14 bean and 16 forage peanut accessions were determined and compared to the classical International Board for Plant Genetic Resources (IBPGR) descriptors. Descriptors discriminating most accessions were identified using a random forest algorithm. The most-valuable classification descriptors for peanuts were 100-seed weight and days to flowering, and for beans, days to flowering and primary seed color. The combination of phenomic and classical descriptors increased the accuracy of the classification of Phaseolus and Arachis accessions. Functional diversity indices are recommended to genebank curators to evaluate phenotypic variability to identify accessions with unique traits or identify accessions that represent the greatest phenotypic variation of the species (functional agrobiodiversity collections). The artificial intelligence algorithms are capable of characterizing accessions which reduces costs generated by additional phenotyping. Even though deep analysis of data requires new skills, associating genetic, morphological and ecogeographic diversity is giving us an opportunity to establish unique functional agrobiodiversity collections with new potential traits.

Self-Esteem and Coping Strategies in Adolescent Cancer Patients during the Period of Illness and Follow-Up.

The importance of self-esteem during the course of oncological illness has been well-documented by some previous studies. However, data assessing its association with various coping strategies, especially considering the period of illness, are still scarce. The objective of this study is to analyze the differences in coping strategies among oncological adolescents, taking into account their self-esteem, illness period, age, and sex. A total of 201 oncological patients between the ages of 12 and 17 from three different Spanish cities were included in this study. All of them were asked to answer a tailored questionnaire, encompassing information about age, sex, and illness period. Additionally, the coping strategies were measured using the ACS scale, while self-esteem was evaluated using the SENA questionnaire. The results demonstrated that male adolescents and older individuals exhibited higher levels of self-esteem. The main coping strategies associated with higher self-esteem were "ignore the problem", "focus on positive", "physical recreation", and "wishful thinking" both during the treatment and the follow-up phases. We conclude that higher self-esteem is associated with some of the coping strategies such as "focus on positive", "ignore the problem", and "wishful thinking". Sociodemographic variables influence the relationship between self-esteem and coping strategies, but no differences were found regarding the period of illness.

Pelleting and particle size reduction of corn increase net energy and digestibility of fiber, protein, and fat in corn-soybean meal diets fed to group-housed pigs.

BACKGROUND: Reduction of the particle size of corn increases energy digestibility and concentrations of digestible and metabolizable energy. Pelleting may also reduce particle size of grain, but it is not known if there are interactions between particle size reduction and pelleting. The objective of this experiment was to test the hypothesis that particle size reduction and pelleting, separately or in combination, increase N balance, apparent total tract digestibility (ATTD) of fiber and fat, and net energy (NE) in corn-soybean meal diets fed to group-housed pigs. METHODS: Six corn-soybean meal-based diets were used in a 3 × 2 factorial design with 3 particle sizes of corn (i.e., 700, 500, or 300 µm) and 2 diet forms (i.e., meal or pelleted). Pigs were allowed ad libitum access to feed and water. Twenty-four castrated male pigs (initial weight: 29.52 kg; standard diviation: 1.40) were allotted to the 6 diets using a 6 × 6 Latin square design with 6 calorimeter chambers (i.e., 4 pigs/chamber) and 6 periods. Oxygen consumption and CO2 and CH4 productions were measured during fed and fasting states and fecal and urine samples were collected. RESULTS: Regardless of particle size of corn, the ATTD of gross energy (GE), N, and acid-hydrolyzed ether extract (AEE), and the concentration of NE were greater (P < 0.05) in pelleted diets than in meal diets. Regardless of diet form, the ATTD of GE, N, and AEE, and the concentration of NE were increased (linear; P < 0.05) by reducing the particle size of corn, but the increase was greater in meal diets than in pelleted diets (interaction; P < 0.05). CONCLUSIONS: Both pelleting and reduction of corn particle size increased nutrient digestibility and NE, but increases were greater in meal diets than in pelleted diets.

Angiographic patterns in spontaneous coronary artery dissection: novel diagnostic insights.

INTRODUCTION: Spontaneous coronary artery dissection (SCAD) is a rare but well-recognized cause of acute coronary syndrome, especially important in women. Invasive coronary angiography (ICA) is the fundamental diagnostic technique for the confirmation of SCAD. Knowing the angiographic patterns suggestive of SCAD is essential for the correct identification of patients with this entity. AREAS COVERED: In this narrative review, the main angiographic characteristics of SCAD lesions as detected by ICA are presented and discussed. EXPERT OPINION: In addition to the specific angiographic classification of SCAD, several authors have described complementary angiographic patterns suggestive of SCAD. Knowledge and correct identification of these angiographic patterns is essential for the correct diagnosis of patients with clinical suspicion of SCAD.

Necroptosis blockade prevents lung injury in severe influenza.

Severe influenza A virus (IAV) infections can result in hyper-inflammation, lung injury and acute respiratory distress syndrome1-5 (ARDS), for which there are no effective pharmacological therapies. Necroptosis is an attractive entry point for therapeutic intervention in ARDS and related inflammatory conditions because it drives pathogenic lung inflammation and lethality during severe IAV infection6-8 and can potentially be targeted by receptor interacting protein kinase 3 (RIPK3) inhibitors. Here we show that a newly developed RIPK3 inhibitor, UH15-38, potently and selectively blocked IAV-triggered necroptosis in alveolar epithelial cells in vivo. UH15-38 ameliorated lung inflammation and prevented mortality following infection with laboratory-adapted and pandemic strains of IAV, without compromising antiviral adaptive immune responses or impeding viral clearance. UH15-38 displayed robust therapeutic efficacy even when administered late in the course of infection, suggesting that RIPK3 blockade may provide clinical benefit in patients with IAV-driven ARDS and other hyper-inflammatory pathologies.

Substitution Effects in Aryl Halides and Amides into the Reaction Mechanism of Ullmann-Type Coupling Reactions.

In this article, we present a comprehensive computational investigation into the reaction mechanism of N-arylation of substituted aryl halides through Ullmann-type coupling reactions. Our computational findings, obtained through DFT ωB97X-D/6-311G(d,p) and ωB97X-D/LanL2DZ calculations, reveal a direct relation between the previously reported experimental reaction yields and the activation energy of haloarene activation, which constitutes the rate-limiting step in the overall coupling process. A detailed analysis of the reaction mechanism employing the Activation Strain Model indicates that the strain in the substituted iodoanilines is the primary contributor to the energy barrier, representing an average of 80% of the total strain energy. Additional analysis based on conceptual Density Functional Theory (DFT) suggests that the nucleophilicity of the nitrogen in the lactam is directly linked to the activation energies. These results provide valuable insights into the factors influencing energetic barriers and, consequently, reaction yields. These insights enable the rational modification of reactants to optimize the N-arylation process.

Development and testing of a sedation protocol for Neocaridina davidi.

Neocaridina davidi, a small freshwater shrimp native to Asia, specifically China, Japan, Korea, and Vietnam, possesses remarkable resistance to poor water quality and offers various advantages over other invertebrate species to examine crucial issues in neuroscience and other related areas. These advantages include robustness, ease of maintenance, and transparency, making them useful for in vivo studies with optical imaging techniques. Despite its suitability for research purposes, particularly in the fields of imaging and fluorescent techniques, the lack of attention given to this species has resulted in the absence of a robust and replicable sedation protocol for immobilization and safe manipulation. Consequently, researchers face challenges in performing experimental procedures while minimizing harm to this specimen. In this study, we have developed and evaluated a simple sedation protocol specifically designed for Neocaridina davidi, assessing its effectiveness using light microscopy and image processing.

Impact of sex differences on the feasibility and safety of distal radial access for coronary procedures: a multicenter prospective observational study.

BACKGROUND: Conventional transradial access in women is associated with a lower success rate and a higher incidence of spasm compared to men. To date, the effect of sex on the performance of distal radial access (DRA) has not been fully elucidated. The aim of this study was to assess the impact of sex on catheterization success and other performance parameters of DRA procedures. METHODS: This is a prospective three-center observational study. From August 2020 to September 2022, data from all consecutive patients who underwent DRA for coronary procedures were collected. RESULTS: A total of 868 procedures were registered and stratified into two groups according to sex: women (n = 258) and men (n = 610). Female patients had less favorable baseline characteristics than male patients in terms of absent or weak pulse (29% vs. 17%; P  < 0.001), distal radial diameter (2.2 ±â€…0.3 vs. 2.4 ±â€…0.4 mm; P  < 0.001) and proximal radial diameter (2.5 ±â€…0.7 vs. 2.7 ±â€…0.7 mm; P  = 0.001). No differences in success rates were found in women compared to men (94.2% vs. 96.6%; P  = 0.135), with a higher presence of arterial spasm in women (5.8% vs. 3.0%; P  = 0.044). The preprocedural ultrasound evaluation was the only predictor of DRA success [odds ratio = 20.0 (4.739-83.333); P  < 0.001]. CONCLUSION: In patients undergoing coronary procedures, the success rate of DRA was high regardless of sex, with a higher incidence of arterial spasm in women.

The Septic Elbow Joint: Treatment Approaches for Improved Patient Outcomes.

Septic arthritis of the elbow is a serious problem requiring prompt, accurate diagnosis and urgent surgical intervention. Achieving successful patient outcomes depends heavily on early diagnosis and efficient streamlined surgical treatment. Essential tactics for treating the septic elbow joint include immediate joint irrigation and debridement in addition to administration of appropriate antibiotics. This comprehensive review delves into the cause of the septic elbow joint, identifies associated risk factors, and provides a comprehensive approach encompassing the diagnosis and treatment of the septic elbow. The aim of this review is to optimize patient care and outcomes.

Mitochondrial DNA competition: starving out the mutant genome.

High levels of pathogenic mitochondrial DNA (mtDNA) variants lead to severe genetic diseases, and the accumulation of such mutants may also contribute to common disorders. Thus, selecting against these mutants is a major goal in mitochondrial medicine. Although mutant mtDNA can drift randomly, mounting evidence indicates that active forces play a role in the selection for and against mtDNA variants. The underlying mechanisms are beginning to be clarified, and recent studies suggest that metabolic cues, including fuel availability, contribute to shaping mtDNA heteroplasmy. In the context of pathological mtDNAs, remodeling of nutrient metabolism supports mitochondria with deleterious mtDNAs and enables them to outcompete functional variants owing to a replicative advantage. The elevated nutrient requirement represents a mutant Achilles' heel because small molecules that restrict nutrient consumption or interfere with nutrient sensing can purge cells of deleterious mtDNAs and restore mitochondrial respiration. These advances herald the dawn of a new era of small-molecule therapies to counteract pathological mtDNAs.