7-day Period Research Articles

Long-Term in vivo Observation of Maize Leaf Xylem Embolism, Transpiration and Photosynthesis During Drought and Recovery.

Plant water transport is essential to maintain turgor, photosynthesis and growth. Water is transported in a metastable state under large negative pressures, which can result in embolism, that is, the loss of function by the replacement of liquid xylem sap with gas, as a consequence of water stress. To avoid experimental artefacts, we used an optical vulnerability system to quantify embolism occurrence across six fully expanded maize leaves to characterize the sequence of physiological responses (photosynthesis, chlorophyll fluorescence, whole-plant transpiration and leaf inter-vein distance) in relation to declining water availability and leaf embolism during severe water stress. Additionally, we characterize the recovery of leaf function in the presence of sustained embolism during a 6-day recovery period. Embolism formation occurred after other physiological processes were substantially depressed and were irreversible upon rewatering. Recovery of transpiration, net CO2 assimilation and photosystem II efficiency were aligned with the severity of embolism, whereas these traits returned to near pre-stress levels in the absence of embolism. A better understanding of the relationships between embolism occurrence and downstream physiological processes during stress and recovery is critical for the improvement of crop productivity and resilience.

Influenza A Virus Subtype H9N2 Infection Induces Respiratory Microbiota Dysbiosis in Chickens via Type-I Interferon-Mediated Mechanisms

Abstract Avian influenza virus (AIV) poses significant threats to poultry and human health. This study investigates the impact of H9N2 AIV infection on the respiratory microbiota of chickens using 16S rRNA gene sequencing. Forty-eight one-day-old specific pathogen-free (SPF) chickens were assigned to six groups: a control and five post-infection groups (days 1, 3, 5, 7, and 9). After a 15-day microbiota stabilization period, the infected chickens received a viral inoculum (10^7 TCID50/mL) via ocular, intra-nasal, and intra-tracheal routes. Tracheal and broncho-alveolar lavage samples were analyzed. Significant reductions in microbiota diversity were observed on days 5, 7, and 9 post-infection, compared to d0 controls. PERMANOVA confirmed significant beta diversity differences (P = 0.001) between infected and uninfected groups. The microbial shifts from d5 to d9 were marked by increased Proteobacteria, decreased Actinobacteria and Firmicutes, and a rise in Dickeya. Elevated type I interferon (IFN-β) and viperin gene expression at d5 coincided with reduced microbiota diversity, highlighting the respiratory microbiota's role in modulating host responses to AIV H9N2 infection and suggesting potential biomarkers for respiratory dysbiosis.

Characterizing Prodrome (Premonitory Phase) in Migraine: Results From the PRODROME Trial Screening Period.

Limited data are available describing the frequency, severity, and consistency of prodromal symptoms followed by headache. This analysis of the PRODROME trial screening period characterized prodromal symptoms in people with migraine, including the most common symptoms and their severity, and the frequency and consistency with which prodromal symptoms were followed by headache. PRODROME was a multicenter, randomized, double-blind, placebo-controlled, crossover trial conducted in the United States that enrolled adults with 2-8 migraine attacks per month who stated they could identify prodromal symptoms that were reliably followed by a headache. The trial included a 60-day screening period designed to test the predictive validity of "qualifying prodrome events" before the onset of headache. Participants used an eDiary to report qualifying prodrome events, defined as prodromal symptoms whereby the participant was confident a headache would follow within 1-6 hours. This analysis evaluated common prodromal symptoms and their severity, time from prodrome onset to headache onset, and the percentage of participants who identified prodromal symptoms that were followed by a headache ≥75% of the time over the 60-day screening period. A total of 920 participants entered eDiary data, with a mean of 5.2 qualifying prodrome events during the 60-day screening period. A total of 4,802 qualifying prodrome events were recorded. The most common prodromal symptoms identified were sensitivity to light (57.2%; 2,748/4,802), fatigue (50.1%; 2,408/4,802), neck pain (41.9%; 2,013/4,802), sensitivity to sound (33.9%; 1,630/4,802), either difficulty thinking or concentrating (30.0%; 1,442/4,802), and dizziness (27.8%; 1,333/4,802). Of all qualifying prodrome events reported, 81.5% (3,913/4,802) were followed by headache of any intensity within 1-6 hours. For each participant, a mean of 84.4% of their qualifying prodrome events were followed by a headache within 1-6 hours, with 76.9% of participants identifying qualifying prodrome events that were followed by headache within 1-6 hours ≥75% of the time. Participants were able to identify migraine attacks in which prodromal symptoms were reliably followed by a headache within 1-6 hours. These findings suggest the potential for initiating treatment during the prodrome to prevent headache. ClinicalTrials.gov NCT04492020. Submitted: July 27, 2020; First patient enrolled: August 21, 2020. clinicaltrials.gov/study/NCT04492020.

An interpretable machine learning tool for in-home monitoring of agitation episodes in people living with dementia: a proof-of-concept study.

Agitation affects around 30% of people living with dementia (PLwD), increasing carer burden and straining care services. Agitation identification typically relies on subjective clinical scales and direct patient observation, which are resource-intensive and challenging to incorporate into routine care. Clinical applicability of data-driven methods for agitation monitoring is limited by constraints such as short observational periods, data granularity, and lack of interpretability and generalisation. Current interventions for agitation are primarily medication-based, which may lead to severe side effects and lack personalisation. Understanding how real-world factors interact with agitation within home settings offers a promising avenue towards identifying potential personalised non-pharmacological interventions. We used longitudinal data (32,896 person-days from n=63 PLwD) collected using in-home monitoring devices between December 2020 and March 2023. Employing machine learning techniques, we developed a monitoring tool to identify the presence of agitation during the week. We incorporated a traffic-light system to stratify agitation probability estimates supporting clinical decision-making, and employed the SHapley Additive exPlanations (SHAP) framework to enhance interpretability. We designed an interactive tool that enables the exploration of personalised non-pharmacological interventions, such as modifying ambient light and temperature. Light Gradient-boosting Machine (LightGBM) achieved the highest performance in identifying agitation over an 8-day period with a sensitivity of 71.32%±7.38 and specificity of 75.28%±7.38. Implementing the traffic-light system for stratification increased specificity to 90.3%±7.55 and improved all metrics. Key features for identifying agitation included low nocturnal respiratory rate, heightened alertness during sleep, and increased indoor illuminance, as revealed by statistical and feature importance analysis. Using our interactive tool, we identified indoor lighting and temperature adjustments as the most promising and feasible intervention options within our cohort. Our interpretable framework for agitation monitoring, developed using data from a dementia care study, showcases significant clinical value. The accompanying interactive interface allows for the in-silico simulation of non-pharmacological interventions, facilitating the design of personalised interventions that can improve in-home dementia care. This study is funded by the UK Dementia Research Institute [award number UK DRI-7002] through UK DRI Ltd, principally funded by the Medical Research Council (MRC), and the UKRI Engineering and Physical Sciences Research Council (EPSRC) PROTECT Project (grant number: EP/W031892/1). Infrastructure support for this research was provided by the NIHR Imperial Biomedical Research Centre (BRC) and the UKRI Medical Research Council (MRC). P.B. is also funded by the Great Ormond Street Hospital and the Royal Academy of Engineering. C.S. is supported by the UK Dementia Research Institute [award number UK DRI-5209], a UKRI Future Leaders Fellowship [MR/MR/X032892/1] and the Edmond J. Safra Foundation. R.N. is funded by UK Dementia Research Institute [award number UK DRI-7002] and the UKRI Engineering and Physical Sciences Research Council (EPSRC) PROTECT Project (grant number: EP/W031892/1). M.B. and A.K.S. are funded by the UK Dementia Research Institute [award number UKDRI-7002 and UKDRI-5209]. N.F.L., A.C., C.W. and S.K.arefunded bythe UK Dementia Research Institute[award number UK DRI-7002].

Rabbit teeth serve as a calcium source for rabbits (Oryctolagus cuniculus).

Rabbits have hypselodont (ever-growing) teeth; dental growth constantly replaces dental tissue lost due to abrasion and attrition. It has been suggested that rabbits must have high mineral requirements, in particular for calcium, to fuel this constant tooth growth. However, this assumption ignores the fact that tooth wear represents finely ground tissue that is not lost to the body but swallowed during food processing. No studies exist regarding the possible recycling of dental enamel and dentin calcium. In this cross-over study, eight-months old intact female rabbits were fed with a complete pelleted diet containing either calcium carbonate/dicalcium phosphate (control) or ground rabbit teeth as calcium sources for 15 days each. After 11 days of adaptation, intake, urinary and faecal losses were quantified in a 4-day period in metabolism cages, and blood mineral levels were determined in a final blood sample. There were few relevant, significant differences between the treatments, with the exception of calcium digestibility (20 ±8 % on the control vs. 33 ±6 % on the tooth meal diet, P = 0.002) and total calcium retention (0.10 ±0.13 vs. 0.33 ±0.10 g/d, P = 0.004). The results suggest that rabbits are well able to recycle the minerals from dental tissue lost during dental wear.

A Cutting-Edge Multilayer Nanofiber Wound Dressing: Design, Synthesis, and Investigation for Enhanced Wound Healing InVitro and InVivo.

Wounds, disruptions in normal anatomy, are classified as acute or chronic. The choice of wound treatment relies significantly on dressing materials. Electrospun nanofibrous materials offer promising applications in wound healing, featuring a substantial surface area, close mimicry of the natural extracellular matrix, and adjustable water resistance, air permeability, and drug release. This research endeavors to formulate an innovative three-layered nanofibrous wound dressing using the electrospinning technique with the primary objectives of enhancing patient well-being, exhibiting antimicrobial characteristics, and expediting wound healing. The designed dressing comprises nanofibers of polyurethane (PU), quercetin (Q)-loaded polyethylene glycol (PEG), polyvinyl alcohol (PVA), and gelatin. Characterization of individual layers and the integrated wound dressing was conducted through SEM and FT-IR analyses. The efficacy of the nanofibrous wound dressing was assessed through invitro human cell culture and invivo rat wound models. The anti-toxic effects of nanofiber wound dressing on human epithelial and keratin cells have been proven. Invitro wound models in 24-well plates were utilized to assess the impact on wound healing rates. Photographic documentation of wound closure was performed at the different treatment hours, revealing complete closure of the wounds by the end of the 48th hour. Rats with 2 × 1 cm wounds were treated with the nanofibrous dressings, and wound healing progress was observed over a 14-day period. qRT-PCR was employed to analyze MMP-9, TIMP1, COL1A1, PDGFA, and VEGFC mRNA expressions. With its contemporary design surpassing existing treatments, the nanofiber wound dressing stands out for its wound-healing acceleration and antibacterial properties.

Impact of Bambusa vulgaris-supplemented diet on Nile tilapia challenged with Pseudomonas putida: Hematological, immune, and oxidative responses.

This study investigated the effects of bamboo shoot extract (Bambusa vulgaris) as a feed additive on the health profiles and infection resistance of Nile tilapia (Oreochromis niloticus) against Pseudomonas putida. Bamboo shoot extract was added at levels of 0g, 40g, and 60g per 1000g of diet over a 60-day period. The fish were then challenged with a pathogenic P. putida strain. Chemical analysis of the bamboo shoot extract identified 3,5-dinitrophenol and hydroquinone as the two most abundant compounds. Results showed that fish fed bamboo-enriched diets exhibited significantly enhanced levels of red blood cells, hemoglobin, hematocrit, white blood cells, and platelets, and improved erythrocyte cellular and nuclear morphologies, indicating improved health profiles after the challenge. Liver function indicators, including AST, ALT, and ALP, were notably balanced in fish receiving bamboo shoot extract post-challenge (p<0.05). Blood levels of K+ were lower in the bamboo-fed groups. Additionally, blood levels of Ca++ and Na+ were reduced in fish fed 40g and 60g of bamboo, respectively, compared to the control group (p<0.01). The bamboo extract also enhanced immune and oxidative capacities, as demonstrated by increased catalase, superoxide dismutase, lysozyme activity, and phagocytic activity, along with reduced malondialdehyde levels and elevated serum immunoglobulin M (p<0.01). Gene expression analysis revealed significant effects of Bambusa vulgaris extract, Pseudomonas infection, and their interaction on the expression of interleukin-1β, interleukin-10, and NK-lysin genes, with varying expression levels at 1, 3, and 7 days post-challenge (p<0.05). The liver bacterial load in fish exposed to P. putida significantly decreased in the bamboo-fed groups, with the lowest count observed in the 60g bamboo group. Additionally, survival rates were markedly higher in the bamboo-fed groups compared to the control, with no significant difference between the two bamboo-fed groups. In conclusion, dietary supplementation with bamboo shoot extract enhances hematological parameters, blood cell and nuclear morphology, and increases survival rates in Nile tilapia following infection.

Clinical outcomes among COVID-19 patients initiated on molnupiravir in Denmark - A national registry study.

Molnupiravir (MOV) is an orally bioavailable ribonucleoside with antiviral activity against all tested SARS-CoV-2 variants. We describe the demographic, clinical, and treatment characteristics of non-hospitalized Danish patients treated with MOV and their clinical outcomes following MOV initiation. Among all adults (>18years) who received MOV between 16 December 2021 and 30 April 2022 in an outpatient setting in Denmark, we summarized their demographic and clinical characteristics at baseline and post-MOV outcomes using descriptive statistics. Outcomes were emergent hospitalization and all-cause mortality during the 28 days after MOV initiation. We estimated the odds ratios (OR) of outcomes by time from positive test to treatment using logistic regression. We identified 3691 MOV-treated patients, of whom 45.8% were male and mean age was 70.1 years. Most patients (76.2%) initiated MOV within 0-2 days after a positive SARS-CoV-2 test and 16.8% within 3-5 days. Over a 28-day period, rates for all-cause, respiratory- or COVID-19-related, and COVID-19-related hospitalization were 4.8%, 2.6% and 1.5%, respectively. All-cause mortality was 1.6%. Initiation of MOV 3-5 days after a positive SARS-CoV-2 test compared to 1-2 days was associated with an increased risk of all-cause (OR 1.85, 95% CI 1.29-2.67) and respiratory or COVID-19-related (OR 1.78, 95% CI 1.07-2.94) hospitalization, and all-cause mortality (OR 2.90, 95% CI 1.64-5.15). MOV was primarily prescribed to vaccinated elderly persons with multiple comorbidities. The all-cause hospitalization and mortality rates in this population were low. Early initiation of MOV reduced the risk of hospitalization and death compared with late initiation.

Size-dependent dynamics and tissue-specific distribution of nano-plastics in Danio rerio: Accumulation and depuration

Nano-plastics (NPs), defined as particles smaller than 1µm, have emerged as a significant environmental contaminant due to their potential ecological impacts. This study explores the size-dependent dynamics and tissue-specific distribution of polystyrene nano-plastics (PS-NPs) in Danio rerio exposed to PS-NPs at an environmentally relevant concentration of 1μg/mL for 28 days, followed by a 17-day depuration period. PS-NPs of 20, 100, 200, and 500nm were assessed in the intestine, liver, gills, muscle, and brain using transmission electron microscopy (TEM) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Smaller PS-NPs (20nm) showed the highest accumulation in the intestine, followed by the liver, and gills, due to their greater surface area and cellular penetration. In contrast, larger PS-NPs (500nm) exhibited lower accumulation and clearance rates, especially in the brain, suggesting restricted passage through biological barriers. The intestine consistently had the highest concentrations in both accumulation and depuration, while the brain maintained the lowest across all nanoparticle sizes. During depuration, smaller particles cleared more quickly, whereas larger particles persisted. This study highlights the tissue-specific distribution and retention patterns of PS-NPs in D. rerio, providing insights into nanoparticle behavior in aquatic organisms and the need for long-term size-specific environmental risk assessments.

Economic Impact of Respiratory Syncytial Virus Infections in Children Under 5 Years of Age Attending Primary Care in Italy: A Prospective Cohort Study in Two Regions.

Accurate cost estimates of respiratory syncytial virus (RSV) infections in primary care are limited, despite the majority of cases being managed in this setting. This study aims to estimate healthcare costs for children with RSV in primary care and the related costs of parental work absence. Children < 5 years of age with symptoms of acute respiratory infections were recruited via primary care paediatricians in two Italian regions for a prospective cohort study on the RSV burden in primary care, during the 2019/2020 winter. Healthcare utilization, medication use and parental work absence were assessed during a 14-day follow-up period. Average costs were estimated per RSV episode for the overall study population, as well as per age group. Two hundred ninety three children were recruited, of which 119 tested RSV positive (41%) and 109 were included. In total, 89% of RSV-positive children (97/109) had ≥ 1 repeat paediatrician visit(s), and 10% (11/109) visited the ED. The mean number of repeat visits was 3.8 (SD: 4.0) and the mean duration of work absence 4.0 days (SD: 5.0). Average costs per RSV episode were €730 (95% CI: €691-€771), with direct medical costs accounting for 25% (€183 [95% CI: €174-€191]) and indirect costs related to work absence for 75% (€547 [95% CI: €509-€587]). Costs associated with RSV infections in young children in primary care are considerable due to a substantial number of paediatrician visits and high rates of parental work absence. These costs are important to include in decision-making regarding the implementation of new RSV immunization strategies in national immunization programmes.

Exploring the "Microbe-Metabolite-Gene" regulatory mechanism of compound probiotics on antioxidant function in heat-stressed broilers.

Nutritional modification strategies have become pivotal in addressing heat stress in poultry farming. Probiotics are increasingly recognized as a sustainable additive by researchers. The enhancement of antioxidant capacity is critical for improving the overall health and productivity of broilers. However, the molecular mechanisms by which probiotics influence antioxidant functions in heat-stressed broilers remain unclear. Consequently, this study raised 400 one day old Arbor Acres broilers until 28 d. Among them, 300 broilers, showing no significant weight differences (P > 0.05), were randomly divided into three groups (control, heat stress, and heat stress with probiotics), each consisting of five replicates with 20 broilers per replicate. The heat stress conditions were maintained at 32 ± 1°C from 9:00 to 17:00, and at 21 ± 1°C during the remaining hours, identical to the control conditions. The heat stress with probiotics group received a basal diet supplemented with 10 g/kg of compound probiotics (Lactobacillus casei: Lactobacillus acidophilus: Bifidobacterium at a ratio of 1:1:2). All groups had ad libitum access to food and water over the 14-day experimental period. Results indicated that the compound probiotics enhanced growth performance and antioxidant capacity, increasing levels of glutathione peroxidase and catalase. Weighted correlation network analysis (WGCNA) identified Hub genes (e.g., PANK2, FAM167A, ABCG8, DYDC2), Hub metabolites (e.g., l-glutamine, ornithine), and Hub microorganisms (e.g., Burkholderia, Macromonas) that regulate antioxidant functions in heat-stressed broilers. By integrating the WGCNA results, we constructed a "microbe-metabolite-gene" regulatory network centered around co-enriched pathways, illustrating the interrelationships between molecules. Notably, NT5C1A, GDA,l-glutamine, and guanosine were notably enriched in purine metabolism, whereas ABCG8,l-arginine, allose, and deoxyguanosine were prominently enriched in ABC transporters pathways, highlighting their crucial involvement in orchestrating the antioxidant response to heat stress. This study elucidates the molecular mechanisms by which compound probiotics enhance antioxidant functions in heat-stressed broilers, offering a theoretical foundation for probiotic applications in poultry.

Dual-acting mechanism of microbial corrosion inhibition by ginger extract against nitrate-reducing bacteria in a nutrient-deficient environment.

This study investigates the efficacy of ginger extract (GE) as a green corrosion inhibitor in the petrochemical industry, specifically targeting microbial corrosion in carbon steel pipes utilized for river water coolant systems. The nutrient-deficient conditions coupled with the presence of nitrate-reducing bacteria (NRB) within these pipelines facilitate microbiologically influenced corrosion, wherein bacteria directly interact with the metal to harvest electrons for metabolic processes. Our findings indicate that GE inhibits microbial corrosion through dual mechanisms: significantly reducing biofilm formation and adsorbing onto the metal surface. Biofilm inhibition was assessed through both qualitative and quantitative assessments, employing light microscopy and confocal laser scanning microscopy (CLSM) to visualize and confirm the suppression of biofilm development by NRB in the presence of GE. Electrochemical techniques, including electrochemical impedance spectroscopy (EIS) and polarization measurements, demonstrated that GE achieved an inhibition efficiency nearing 95%. Further analysis via scanning electron microscopy (SEM) and adsorption studies corroborated the effective adsorption of GE on carbon steel, highlighting the importance of concentration and immersion time in performance outcomes. Notably, elevated concentrations of GE were found to enhance inhibitory effects, with a 7-day exposure period yielding optimal surface coverage. These results establish GE as a promising and environmentally friendly alternative for mitigating microbial corrosion in industrial applications. PRACTITIONER POINTS: The presence of nitrate-reducing bacteria (NRB) and a nutrient-starved environment are commonplace in the coolant pipes carrying water and can cause severe damage to carbon steel pipes. Ginger extract (GE), an inexpensive green corrosion inhibitor, can be a potent microbial corrosion inhibitor in petrochemical industries. The high efficiency (95%), achieved with ginger extract (GE), is due to its dual-acting mechanism: It inhibits the formation of bacterial biofilm on the surface of carbon steel by providing a non-conducive environment for bacterial growth. The components of GE get adsorbed over the surface of carbon steel and hence prevent bacterial attachment. An optimal concentration of 0.5g/L GE was required to exhibit high efficiency (95%), which can be achieved on 7days of exposure of carbon steel to GE, in the presence of NRB in a nutrient-starved environment.

Influence of Food Pigments and Thermal Aging on the Color Stability of Denture Base Resins

Color stability of acrylic resins is essential for preserving the aesthetic appearance of denture bases over time. This study explores how food pigments and thermal changes affect the color stability of commonly used denture base resins. Four acrylic resins were tested: three heat-cured acrylic resins with different characteristics (Zhermack® Villacryl H Plus V2, H Plus V4, and H Rapid FN V4) and one self-cured acrylic resin (Zhermack® Villacryl S V4). To simulate the oral environment, the resins underwent 1000 thermal cycles between 5 °C and 55 °C, followed by a 7-day immersion period in beverages such as coffee, red wine, a caramel-colored soft drink (cola), and distilled water (control), forming sixteen group of specimens (n = 5). Color changes (∆E) were measured using the VITA Easyshade V® spectrophotometer, following the CIEDE2000 standard. The findings revealed that thermal aging caused noticeable color changes in all resins (p &lt; 0.001). Red wine led to the most intense discoloration, followed by coffee. The caramel-colored soft drink caused moderate staining, while distilled water had a negligible effect. The type of polymerization did not affect the degree of discoloration, as no significant differences were found between the resins after exposure to beverages (p &gt; 0.05). Overall, this study highlights how both internal and external factors impact the appearance of acrylic resins. Thermal aging can accelerate polymer degradation, while pigments in beverages cause visible staining. Among the tested beverages, red wine proved to be the most aggressive due to its high pigment concentration and low pH. These findings emphasize the need for improved material formulations to enhance the longevity and aesthetic performance of dentures.

Clinical results of a phase 1 trial evaluating a HER2 directed CAR-T product selective for the tumor microenvironment (TME) in patients with GI and other solid tumor malignancies.

449 Background: A major limitation of CAR-T in solid tumors has been the inability to safely target antigens shared between healthy and malignant tissue. One approach to improving selectivity is to regulate CAR affinity through the unique metabolism of the tumor microenvironment (TME). This first in human trial evaluated an autologous (41BB) CAR-T product encoding a tumor metabolism regulated HER2 CAR. Methods: This Phase 1, dose escalation, basket trial evaluated the safety, tolerability, pharmacokinetics and efficacy in 12 patients (pts) with HER2 IHC 3+ / FISH + Stage IV r/r solid tumor malignancies (gastric, esophageal, colorectal, breast). They had failed multiple lines of therapy (66.7% patients had failed 4~10 lines of therapies previously). Dose cohorts of 3E5 (n=3), 1E6 (n=3), or 1E7 (n=6) CAR-T cells/kg were evaluated with a 28-day in-patient DLT observation period. Cell product was manufactured from whole blood, with a 12-day closed process and release on day 17. Lymphodepletion regimen was 500 mg/m 2 Cy and 30 mg/kg 2 Flu X 3 prior to infusion of CAR-T. Results: Data cutoff 09/16/24. All 12 patients were evaluable for safety and efficacy. No DLTs, ICANS or SAEs were observed. For HER2 positive normal tissues of special concern (e.g. cardiomyocytes), longitudinal functional, biochemical and clinical cardiovascular tests showed no signs of OTOT. One patient experienced Grade 2 CRS and recovered. No AE’s lead to study discontinuation. In one patient, there were two Grade 3 AEs attributed to product (neutropenia and hypoxia); both which resolved. With a median follow-up of 302 days for cohort 3, 12-month survival rate in this cohort is 83.3%, compared to the 12-month survival rate of 33.3% for cohorts 1 and 2 combined. Encouraging signals of efficacy were observed with an ongoing (at 24 wks) deep PR (100% reduction SLD) in a GC patient. The median peak copy number of CAR-T cells in peripheral blood (n=12) was 1,818 copies/µg DNA (range 403 – 10,466). Conclusions: The tumor metabolism regulated HER2 CAR-T product was generally well tolerated, with no DLTs, and evidence of radiologic objective response in patients with gastroesophageal cancer. Combined, the safety and activity observed in this study suggests that a previously challenging solid tumor antigen may be safely targeted by a CAR with affinity regulated by the metabolism of the TME. Enrollment for the study was complete as of 28-Feb-2024 (NCT04511871). Clinical trial information: NCT04511871 .

Maternal milk cell components are uptaken by infant liver macrophages via extracellular vesicle mediated transport.

Milk is a multifaceted biofluid that is essential for infant nutrition and development, yet its cellular and bioactive components, particularly maternal milk cells, remain understudied. Early research on milk cells indicated that they cross the infant's intestinal barrier and accumulate within systemic organs. However, due to the absence of modern analytical techniques, these studies were limited in scope and mechanistic analysis. To overcome this knowledge gap, we have investigated the transintestinal transport of milk cells and components in pups over a 21-day period. Studies employed a mT/mG foster nursing model in which milk cells express a membrane-bound fluorophore, tdTomato. Using flow cytometry, we tracked the transport of milk cell-derived components across local and systemic tissues, including the intestines, blood, thymus, mesenteric lymph nodes, and liver. These experiments identified milk-derived fluorescent signals in intestinal epithelial and immune cells as well as liver macrophages in 7-day-old pups. However, the minute numbers of macrophages in mouse milk suggest that maternal cells are not systemically accumulating in the infant; instead, pup macrophages are consuming milk cell membrane components, such as apoptotic bodies or extracellular vesicles (EVs). Exvivo experiments using primary macrophages support this hypothesis, showing that immune cells preferentially consumed EVs over milk cells. Together, these data suggest a more complex interplay between milk cells and the infant's immune and digestive systems than previously recognized and highlight the need for future research on the role of milk cells in infant health.

Effects of fluconazole and amphotericin B in the control of cryptococcal pneumonia in vivo

Cryptococcal pneumonia is a severe fungal infection of the respiratory system, predominantly caused by Cryptococcus neoformans. Its incidence is increasing, driven by evolving pathogen dynamics and heightened susceptibility among patient populations. This investigation aimed to assess the combined therapeutic efficacy of Fluconazole and Amphotericin B for cryptococcal pneumonia and to explore the roles of miR-15b and TGF-β1 in modulating treatment response. Twenty-eight patients diagnosed with cryptococcal pneumonia were randomly allocated to receive either Amphotericin B monotherapy (control group) or a combination of Amphotericin B and Fluconazole (observation group) over a 14-day period. Key respiratory function indices—forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and FEV1/FVC ratios—were measured pre- and post-treatment, alongside levels of procalcitonin (PCT), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), miR-15b, and TGF-β1. Results demonstrated marked improvements in pulmonary function within the observation group, with significantly higher FEV1, FVC, and FEV1/FVC values than those in the control group (P < 0.05). Additionally, the observation group exhibited greater reductions in PCT and sTREM-1, coupled with notable elevations in miR-15b and TGF-β1 levels. The combination therapy achieved a total response rate of 92.86%, significantly surpassing the control’s 57.14% efficacy (P < 0.05). These findings indicate that Fluconazole combined with Amphotericin B not only enhances clinical efficacy by mitigating inflammation but also supports lung function recovery with a favourable safety profile, highlighting its utility in managing cryptococcal pneumonia effectively.

Assessment of the nutritional and effluent properties of potential fish-meal-free diets for rainbow trout (Oncorhynchus mykiss W.) in Iran

Research is needed on alternative ingredients for aquafeeds due to rising demand and limited fish meal availability. This study evaluated fish-meal–free diets for rainbow trout in two phases. The first phase focused on assessing the growth performance and nutrient digestibility of six cold-pelleted diets. Aside from the control diet, the test diets contained poultry protein concentrate, blood meal, low-ash poultry by-product meal, 20–40% canola meal, and feather meal (Goldmehl®). Juvenile fish, averaging 5.3 g, were fed the diets for 56 days in a random-block design with four replicates per feed. Feces were collected using the settling method to evaluate nutritional digestibility and phosphorus availability. The feed from the primary phase, which resulted in the least-cost feed conversion ratio, was extruded and fed to fingerlings (14.6 g) in the second phase. This trial added to the first run in measuring digestibility parameters over a 28-day period. In parallel, extruded feed was given to grow-out fish, averaging 172.5 g, for 55 days in triplicate completely randomized tanks to evaluate growth performance and nutrient effluents. Throughout all the experimental runs, a casein-based laboratory pelleted feed was used for the respective controls. Feeding rainbow trout with 35% canola meal in both pelleted and extruded forms showed comparable results (P > 0.05) to the control in terms of growth performance and apparent digestibility for lipid (> 91%), crude protein (> 87%), and organic matter (> 78%). The extruded feed in this study represents an eco-friendly option for the growing aquafeed sector.

Effects of Maternal Probiotics and Piglet Dietary Tryptophan Level on Gastric Function Pre- and Post-Weaning

Knowledge of how novel antigens or dietary stimuli affect stomach development and function in pigs remains limited. This study aimed to investigate stomach characteristics, parietal cell numbers, and the expression of genes essential to the functioning of the fundic and pyloric gland regions at weaning compared to seven days post-weaning and to examine whether maternal probiotic supplementation or piglet dietary tryptophan (Trp) levels influence these stomach parameters. This study has a 2 × 3 factorial design, with 48 sows assigned to one of two diets: basal or basal supplemented with Bacillus subtilis and Bacillus amyloliquefaciens. Their litters received creep diets containing 0.22, 0.27, or 0.33% standardized ileal digestible (SID) Trp. In total, 96 pigs were sacrificed for gastric sampling, 48 on the day of weaning and 48 on day 7 post-weaning. At 7 days post-weaning, pigs had an increased number of parietal cells and expression of parietal cell activity and digestive enzyme (PGA5 and CHIA) genes in the fundic gland region (p &lt; 0.05), although the expression of signaling molecules involved in the regulation of acid secretion was unchanged in the fundic gland region (p &gt; 0.05) and reduced in the pyloric gland region (p &lt; 0.05), compared to the day of weaning. Overall, maternal probiotic supplementation had a significant impact on gene expression in the fundic gland region of the offspring, elevating several genes related to parietal cell activity (CLIC6, HRH2, KCNE1, KCNQ1, CHRM3, CCKBR, and SSTR2) (p &lt; 0.05). Additionally, there were time × maternal interactions, where certain acid secretion pathway (ATP4A and HDC), chitinase enzyme (CHIA), and ghrelin (GHRL) genes were increased in offspring from probiotic sows compared to control sows at weaning (p &lt; 0.05), but not at 7 days post-weaning (p &gt; 0.05). Maternal probiotic supplementation did not influence growth performance pre-weaning or during the 7-day post-weaning period. There was a limited effect of creep Trp level or maternal × creep interactions on performance, gene expression, or parietal cell counts. Low pre-weaning creep intake may have confounded this analysis. In conclusion, maternal probiotic supplementation accelerated the maturation of the offspring’s stomach, particularly in terms of the expression of genes linked to acid secretion from parietal cells.

Time-resolved compositional and dynamics analysis of biofilm maturation and dispersal via solid-state NMR spectroscopy

Dispersal plays a crucial role in the development and ecology of biofilms. While extensive studies focused on elucidating the molecular mechanisms governing this process, few have characterized the associated temporal changes in composition and structure. Here, we employed solid-state nuclear magnetic resonance (NMR) techniques to achieve time-resolved characterization of Bacillus subtilis biofilms over a 5-day period. The mature biofilm, established within 48 h, undergoes significant degradation in following 72 h. The steepest decline of proteins precedes that of exopolysaccharides, likely reflecting their distinct spatial distribution. Exopolysaccharide sugar units display clustered temporal patterns, suggesting the presence of distinct polysaccharide types. A sharp rise in aliphatic carbon signals on day 4 probably corresponds to a surge in biosurfactant production. Different dynamic regimes respond differently to dispersal: the mobile domain exhibits increased rigidity, while the rigid domain remains stable. These findings provide novel insights and perspectives on the complex process of biofilm dispersal.

641. Real-Time Whole Genome Sequencing Surveillance of Healthcare Associated SARS-CoV-2, Respiratory Syncytial Virus (RSV) and Influenza

Abstract Background Seasonal respiratory viruses contribute to substantial healthcare burden, marked by elevated rates of hospitalizations, morbidity and mortality. The transmission dynamics of respiratory viruses in adult acute care hospitals are not well described. To address this, we performed whole genome sequencing (WGS) surveillance to investigate transmission of SARS-CoV-2, RSV, and influenza among hospitalized patients in a single health system. Methods From August 30, 2023 to March 31, 2024, nasal swab specimens positive for SARS-CoV-2, influenza, or RSV were collected weekly from inpatients hospitalized for ≥ 3 days from two hospitals. Additional specimens collected from the same patient within a 14-day period were excluded. Specimens with qPCR cycle threshold (Ct) values ≥ 34, ≥ 39, and ≥ 31 for SARS-CoV-2, RSV and influenza, respectively, were excluded from WGS. WGS was performed using tiled PCR amplicons or Twist Bioscience hybridization method, followed by bioinformatic analyses to assess genetic relatedness [≤ 2 single nucleotide polymorphisms (SNPs) except for influenza (≤ 3 SNPs)]. Clusters were further investigated for epidemiological connections (shared locations or staff). Results We collected 179 SARS-CoV-2, 37 influenza and 16 RSV specimens, of which 93 specimens did not meet the inclusion and the qPCR QC criteria. We sequenced 84% (87/104) of SARS-CoV-2, 86% (19/22) of influenza, and 92% (12/13) of RSV specimens. Of these, 45% (39/87) of SARS-CoV-2 and 32% (6/19) of influenza genomes formed genetically related clusters, each with 2-4 patients. We identified an epidemiological link for 73% (11/15) of SARS-CoV-2 and 100% (2/2) of influenza clusters (Fig. 1 &amp; Fig. 2). For RSV genomes, we identified one cluster with two epidemiologically linked patients (Table1). On average, the transmission clusters spanned a duration of 15 days (range: 1−35 days, Table1). Conclusion Respiratory viral transmission is frequent in hospitals, with around 40% of sequenced specimens genetically related to each other. While this study serves as an initial step towards possible implementation of real-time WGS surveillance, further investigation is needed to address broader implications, including implementation of infection prevention strategies to address transmission clusters. Disclosures Graham M. Snyder, MD, SM, Infectious Diseases Connect: Advisor/Consultant Alexander Sundermann, DrPH, CIC, FAPIC, OpGen: Honoraria Lee Harrison, MD, GSK: Advisor/Consultant|Merck: Advisor/Consultant|Pfizer: Advisor/Consultant|Sanofi: Advisor/Consultant