Functional Studies Research Articles

Potential Association of The Pathogenic Kruppel-like Factor 14 (KLF14) and Adiponectin (ADIPOQ) SNVs with Susceptibility to T2DM.

To evaluate the associations of the pathogenic variants in Kruppel-like Factor 14 (KLF 14) and Adiponectin (ADIPOQ) with susceptibility to type 2 diabetes mellitus (T2DM). Type 2 diabetes mellitus (T2DM) is a pandemic metabolic disease characterized by increased blood sugar and caused by resistance to insulin in peripheral tissues and damage to pancreatic beta cells. Kruppel-like Factor 14 (KLF-14) is proposed to be a regulator of metabolic diseases, such as diabetes mellitus (DM) and obesity. Adiponectin (ADIPOQ) is an adipocytokine produced by the adipocytes and other tissues and was reported to be involved in T2DM. To study the possible association of the KLF-14 rs972283 and ADIPOQ-rs266729 with the risk of T2DM in the Saudi population. We have evaluated the association of KLF-14 rs972283 C>T and ADIPOQ-rs266729 C>G SNV with the risk to T2D in the Saudi population using the Amplification Refractory Mutation System PCR (ARMS-PCR), and blood biochemistry analysis. For the KLF-14 rs972283 C>T SNV we included 115 cases and 116 healthy controls, and ADIPOQ-rs266729 C>G SNV, 103 cases and 104 healthy controls were included. Results indicated that the KLF-14 rs972283 GA genotype and A allele were associated with T2D risk with OR=2.14, p-value= 0.014 and OR=1.99, p-value=0.0003, respectively. Results also ADIPOQ-rs266729 CG genotype and C allele were associated with an elevated T2D risk with an OR=2.53, p=0.003 and OR=1.66, p-value =0.012, respectively. We conclude that SNVs in KLF-14 and ADIPOQ are potential loci for T2D risk. Future large-scale studies to verify these findings are recommended. These results need further verifications in protein functional and large-scale case control studies before being introduced for genetic testing.

Quantification and Functional Studies of Neutrophilic Cells Identifies Distinct Papilloma Phenotypes.

To characterize the distribution of immune cell subsets within laryngeal papillomas and to study the function of potentially immunosuppressive neutrophilic and regulatory T cells (Tregs). Fresh clinical papilloma specimens were collected at the time of surgery and studied with multiparameter flow cytometry. Papilloma infiltrating neutrophilic cells and Tregs were sorted and studied functionally with ex vivo T cell suppression assays. Flow cytometric analysis of fresh laryngeal papillomas samples from 18 adult patients with recurrent respiratory papillomatosis revealed patterns in immune constituency between patients. Clearly divergent phenotypes based primarily on the degree of neutrophilic and T cell infiltration were identified. Relative neutrophilic cell enrichment and T cell depletion were observed in 50% of samples and neutrophilic cell depletion and T cell enrichment were observed in the others. Greater papilloma neutrophilic cell enrichment was positively associated with the number of clinically indicated interventions required in the 12 months prior to sample collection, linking papilloma neutrophil inflammation to disease severity. Functional assays revealed the ability of both papilloma infiltrating neutrophilic and Tregs to suppress T cell function at roughly equal magnitudes, but substantially increased infiltration of neutrophilic cells compared to Tregs across samples. Neutrophilic cells are an important contributor to immunosuppression within the respiratory papilloma microenvironment. Given these data and the association between greater neutrophilic cell infiltration and lack of clinical response to therapeutic vaccination, additional study of strategies aimed at limiting neutrophilic cell infiltration or function within papillomas is warranted. 4 Laryngoscope, 134:3238-3244, 2024.

From Hysteria to Functional Neurological Symptom Disorder: Developments in Clinical Diagnosis and Neurobiology

Changes in the nomenclature of functional neurological symptom disorders (FND) from the past to the present represent historical changes in understanding etiology. Today, there is still difficulty in excluding potential underlying neurological disorders. In addition, there is no consensus on the psychological mechanism leading to the disorder. As a result, diagnostic problems continue to exist. While functional neuroimaging studies show that suppression and conversion mechanisms, which are the concepts of the psychoanalytical theory, may have neural counterparts, neurobiological data suggests that the conversion model cannot be explanatory for every patient. The dorsolateral prefrontal cortex (dlPFC), amygdala, temporoparietal junction (TPJ), insula, anterior cingulate structures, and their connections come to the fore. The fact that the connections between the dlPFC and the hippocampus can prevent the recall of an unwanted memory, as well as the changes detected in the amygdala in these disorders and the increased connectivity between the amygdala and the motor areas, suggest an abnormal connection between emotions and the motor system. It is addressed how changes in the TPJ are related to the loss of the sense of agency. However, it is unclear whether the findings of these studies suggest a "predisposition", "onset of disorder", or "compensatory changes secondary to disorder". Exploring FND to learn how the brain and mind react to psychosocial stressors can be a turning point in understanding the brain-mind connection. The goal of this review is to present the history of the changes in terminology and perspective on this disorder that followed the establishment of psychoanalysis, as well as what kind of evidence has been presented regarding hysteria in light of advances in neuroscience

Roles of the Default Mode Network in Different Aspects of Self-representation When Remembering Social Autobiographical Memories.

Autobiographical memory (AM) is episodic memory for personally experienced events, in which self-representation is more important than that in laboratory-based memory. Theoretically, self-representation in a social context is categorized as the interpersonal self (IS) referred to in a social interaction with a person or the social-valued self (SS) based on the reputation of the self in the surrounding society. Although functional neuroimaging studies have demonstrated the involvement of the default mode network (DMN) in self-representation, little is known about how the DMN subsystems contribute differentially to IS-related and SS-related AMs. To elucidate this issue, we used fMRI to scan healthy young adults during the recollection of AMs. We performed multivariate pattern analysis (MVPA) and assessed functional connectivity in the DMN subsystems: the midline core, medial temporal lobe (MTL), and dorsomedial pFC (dmPFC) subsystems. The study yielded two main sets of findings. First, MVPA revealed that all DMN subsystems showed significant classification accuracy between IS-related and nonsocial-self-related AMs, and IS-related functional connectivity of the midline core regions with the retrosplenial cortex of the MTL subsystem and the dmPFC of the dmPFC subsystem was significant. Second, MVPA significantly distinguished between SS-related and nonsocial-self-related AMs in the midline core and dmPFC subsystems but not in the MTL subsystem, and SS-related functional connectivity with the midline core regions was significant in the temporal pole and TPJ of the dmPFC subsystem. Thus, dissociable neural mechanisms in the DMN could contribute to different aspects of self-representation in social AMs.

Mechanism of Z-Selective Allylic Functionalization via Thianthrenium Salts.

A detailed mechanistic study of the Z-selective allylic functionalization via thianthrenium salts is presented. Kinetic analyses, deuterium labeling experiments, and computational methods are used to rationalize the observed reactivity and selectivity. We find that the reaction proceeds via a rate-determining and stereodetermining allylic deprotonation of an alkenylthianthrenium species. The Z-configuration of the resultant allylic ylide is translated into the Z-allylic amine product through a sequence of subsequent fast and irreversible steps: protonation to form a Z-allylic thianthrenium electrophile and then regioselective substitution by the nucleophile. In the stereodetermining deprotonation step, computational studies identified a series of stabilizing nonbonding interactions in the Z-alkene-forming transition state that contribute to the stereoselectivity.

Hawthorn (Crataegus pinnatifida) berries ripeness induced pectin diversity: A comparative study in physicochemical properties, structure, function and fresh-keeping potential

The effect of hawthorn berries ripeness on the physicochemical, structural and functional properties of hawthorn pectin (HP) and its potential in sweet cherry preservation were investigated. With the advanced ripeness of hawthorn berries, the galacturonic acid (GalA) content decreased from 59.70 mol% to 52.16 mol%, the molecular weight (Mw) reduced from 368.6 kDa to 284.3 kDa, the microstructure exhibited variable appearance from thick lamella towards porous cross-linked fragment, emulsifying activity and emulsions stability, antioxidant activities, α-amylase and pancreatic lipid inhibitory capacities significantly increased. The heated emulsion stored for 30 d presented higher creaming index and more ordered oil droplets compared to the unheated emulsion. With the extended berries ripeness, the firmness of HP gels remarkably decreased from 225.69 g to 73.39 g, while the springiness increased from 0.78 to 1.16, HP exhibited a superior inhibitory effect in water loss, browning, softening, and bacterial infection in sweet cherries preservation.

Non-Equilibrium Assembly of Atomically-Precise Copper Nanoclusters.

Accurate structure control in dissipative assemblies (DSAs) is vital for precise biological functions. However, accuracy and functionality of artificial DSAs are far from this objective. Herein, a novel approach is introduced by harnessing complex chemical reaction networks rooted in coordination chemistry to create atomically-precise copper nanoclusters (CuNCs), specifically Cu11(µ9-Cl)(µ3-Cl)3L6Cl (L = 4-methyl-piperazine-1-carbodithioate). Cu(I)-ligand ratio change and dynamic Cu(I)-Cu(I) metallophilic/coordination interactions enable the reorganization of CuNCs into metastable CuL2, finally converting into equilibrium [CuL·Y]Cl (Y = MeCN/H2O) via Cu(I) oxidation/reorganization and ligand exchange process. Upon adding ascorbic acid (AA), the system goes further dissipative cycles. It is observed that the encapsulated/bridging halide ions exert subtle influence on the optical properties of CuNCs and topological changes of polymeric networks when integrating CuNCs as crosslink sites. CuNCs duration/switch period could be controlled by varying the ions, AA concentration, O2 pressure and pH. Cu(I)-Cu(I) metallophilic and coordination interactions provide a versatile toolbox for designing delicate life-like materials, paving the way for DSAs with precise structures and functionalities. Furthermore, CuNCs can be employed as modular units within polymers for materials mechanics or functionalization studies.

Maximum tumor diameter and renal function can predict the declining surface dose rate after 177Lu-Dotatate: preliminary results of single institution in Japan.

This study aimed to develop a user-friendly prediction formula for dose rate adjustment after initial 177Lu-Dotatate therapy from a prospective observational study of patients. This study included consenting patients in a prospective observational study who underwent their firsttreatment in four cycles of 177Lu-Dotatate treatment at our hospital between January 2022 and February 2024. All patients received 7.4GBq of 177Lu-Dotatate. The prediction formula was derived from the regression analysis of tumor-related factors and renal function. Creatinine clearance was estimated using the Cockcroft-Gault equation in this study for renal function. Among the 13 patients (seven males, six females, median age: 59years), logarithmically transformed total tumor volume (cc) and maximum tumor diameter (mm) of primary tumors or metastases showed strong correlations (p < 0.001, R2 = 0.897). As such, the maximum tumor diameter was used as the tumor parameter in the prediction formula. Additionally, maximum tumor diameter and creatinine clearance showed strong (p < 0.001, R2 = 0.768) and moderate (p = 0.013, R2 = 445) correlations, respectively, with the ratio of the dose rate 5.5-h post-administration to the dose rate immediately post-administration (%) at 1m from the body surface. The resulting formula, 51.4 + 0.360 × maximum tumor diameter (mm) - 0.212 × creatinine clearance (ml/min), demonstrated an extremely strong correlation (p < 0.001, R2 = 0.937). The present study showed that the maximum tumor diameter and renal function affected the declining the dose rate of patients surface after 177Lu-Dotatate, which can inform post-administration dose rate management and potentially facilitate outpatient treatment in Japan.

Viable protoplast isolation, organelle visualization and transformation of the globally distributed plant pathogen Phytophthora cinnamomi.

Phytophthora cinnamomi is an oomycete plant pathogen with a host range of almost 5000 plant species worldwide and therefore poses a serious threat to biodiversity. Omics technology has provided significant progress in our understanding of oomycete biology, however, transformation studies of Phytophthora for gene functionalisation are still in their infancy. Only a limited number of Phytophthora species have been successfully transformed and gene edited to elucidate the role of particular genes. There is a need to escalate our efforts to understand molecular processes, gene regulation and infection mechanisms of the pathogen to enable us to develop new disease management strategies. The primary obstacle hindering the advancement of transformation studies in Phytophthora is their challenging and unique nature, coupled with our limited comprehension of why they remain such an intractable system to work with. In this study, we have identified some of the key factors associated with the recalcitrant nature of P. cinnamomi. We have incorporated fluorescence microscopy and flow cytometry along with the organelle-specific dyes, fluorescein diacetate, Hoechst 33342 and MitoTracker™ Red CMXRos, to assess P. cinnamomi-derived protoplast populations. This approach has also provided valuable insights into the broader cell biology of Phytophthora. Furthermore, we have optimized the crucial steps that allow transformation of P. cinnamomi and have generated transformed isolates that express a cyan fluorescent protein, with a transformation efficiency of 19.5%. We therefore provide a platform for these methodologies to be applied for the transformation of other Phytophthora species and pave the way for future gene functionalisation studies.

Heteroatom Sr doped MCM-41 molecular sieve for VOC adsorption: Study of the surface functionalization and adsorption performance

Volatile organic compounds (VOCs) are recognized as greater adverse effects on both environment and human health, and adsorption is a simple and effective method. Ordered mesoporous MCM-41 molecular sieve was functionalized with heteroatom Sr to change the pore size and surface property by one-step hydrothermal method and adsorption performance for toluene (C7H8) and formaldehyde (CH2O) was investigated. It is found that Sr doped MCM-41 achieves high static adsorption capacities of 534 mg/g for C7H8 and 388 mg/g for CH2O. The dynamic adsorption of C7H8 follows the Langmuir model over Sr doped and undoped MCM-41 under different process conditions, but the dynamic adsorption capacity is only 21 ∼ 99 mg/g. Sr doped MCM-41 maintains a typical pore structure of MCM-41, with some Sr entering the skeleton to form Sr-O-Si bonds. Sr-M−130 has 857 m2/g of SBET and 0.86 cm3/g of Vtotal, but high hydrothermal temperature (150 °C) causes the sintering of MCM-41 structure. Sr doping decreases surface acidity of MCM-41 and increases surface hydroxyl groups (Si-OH). The kinetic diameter and polarity of the adsorbate cause differences in adsorption performance of Sr doped or undoped MCM-41, meaning that the adsorption behavior of non-polar C7H8 mainly depends on surface area and micropore volume while that of polar CH2O is related to the number of ultra-micropore (<1 nm) and forms chemical adsorption of C = O with Si-OH by hydrogen bonds.

My Brain Made Me Do It: The Neuroscience Behind Behavior

Behavior is communication. Although behavior is often interpreted as intentional, examining the biological reasons driving behavior allows healthcare providers to equitably address basic needs, quality of life, and create person-centered care plans. Behaviors in older adults with neurocognitive challenges have great impact on quality of life and healthcare provision. Over 75% of people with dementia experience behavioral disturbances especially as the disease progresses (Press & Alexander, 2019) The healthcare burden is unimaginable including caregiver stress, loss of function in people with neurocognitive illness, and unneeded hospital stays. Despite behavior being the primary way people with advanced neurological illness communicate, healthcare providers use subjective reasoning to assign meaning to behavior. The neurologic basis for behavior is ignored and patient-centered care diminishes. By examining neurologic foundations and connecting it to function, health care providers can apply evidence-based interventions to understand, validate, and provide interventions for behavioral disturbances. This session will provide tools for learners to examine common neurologic behavioral disturbances such as pain, aggression, impulsivity, and delusions in an objective way. By redefining behaviors health care provides can provide strength-based interventions and make informed pharmacological and nonpharmacological care plan decisions. Session will include an overview of functional neuroanatomy, clinical applications, intervention ideas, and case studies.

Tissue-specific reprogramming leads to angiogenic neutrophil specialization and tumor vascularization in colorectal cancer.

Neutrophil (PMN) tissue accumulation is an established feature of ulcerative colitis (UC) lesions and colorectal cancer (CRC). To assess the PMN phenotypic and functional diversification during the transition from inflammatory ulceration to CRC we analyzed the transcriptomic landscape of blood and tissue PMNs. Transcriptional programs effectively separated PMNs based on their proximity to peripheral blood, inflamed colon, and tumors. In silico pathway overrepresentation analysis, protein-network mapping, gene signature identification, and gene-ontology scoring revealed unique enrichment of angiogenic and vasculature development pathways in tumor-associated neutrophils (TANs). Functional studies utilizing ex vivo cultures, colitis-induced murine CRC, and patient-derived xenograft models demonstrated a critical role for TANs in promoting tumor vascularization. Spp1 (OPN) and Mmp14 (MT1-MMP) were identified by unbiased -omics and mechanistic studies to be highly induced in TANs, acting to critically regulate endothelial cell chemotaxis and branching. TCGA data set and clinical specimens confirmed enrichment of SPP1 and MMP14 in high-grade CRC but not in patients with UC. Pharmacological inhibition of TAN trafficking or MMP14 activity effectively reduced tumor vascular density, leading to CRC regression. Our findings demonstrate a niche-directed PMN functional specialization and identify TAN contributions to tumor vascularization, delineating what we believe to be a new therapeutic framework for CRC treatment focused on TAN angiogenic properties.

Unraveling Bulk versus Surface Passivation Effects in Highly Efficient p–i–n Perovskite Solar Cells Using Thiophene‐Based Cations

Defect passivation is nowadays considered a must‐have route for high‐efficiency perovskite solar cells. However, a general rule that correlates the choice of passivating agents with performance enhancements is still missing. Herein, two different thiophene salts that are used as passivating agents are compared, namely thiophene methylammonium chloride and thiophene ethylammonium chloride (TEACl), which are used for the passivation of bulk and surface defects in triple‐cation‐based metal halide perovskites. First, it is observed that the surface passivation method leads to better device performances reaching a power conversion efficiency of 23.56%, with reduced voltage losses and increased fill factor when compared with the reference. Second, it is demonstrated that the chemical structure of the cation dictates its capability either in passivating bulk defects effectively or to form a superficial two‐dimensional/three‐dimensional heterostructure, which happens only for the TEACl case. The chemical composition and the cation dimension are responsible for device performance enhancement as observed by a joint spectroscopic and density functional theory simulations study, providing rational guidelines for further smart device design.

Toward Atomistic Understanding of Iron Phosphate Glass: A First-Principles-Based Density Functional Theory Modeling and Study of Its Physical Properties.

Iron phosphate glasses (IPGs) have been proposed as futuristic materials for nuclear waste immobilization and anode materials for lithium batteries. Recently, many attempts have been made to propose atomistic models of IPGs to explain their properties from an atomistic viewpoint. In this paper, we seek to produce small scale models of IPG that can be handled within the scheme of density functional theory (DFT) to study its electronic structure. The starting models, generated using the Monte Carlo (MC) method, were subsequently annealed using ab initio molecular dynamics (AIMD) to minimize the coordination defects. The geometry of the equilibrated structure was then optimized at 0 K. This hybrid approach (MC + AIMD + DFT optimization) produced good atomistic models of IPG, which can reproduce the experimentally observed band gap, vibrational density of states, magnetic moment of Fe, and mechanical and optical properties. Computationally expensive melt-quench simulation can be avoided using the present approach, allowing the use of DFT for accurate calculations of properties.

Neural correlates of increased alcohol demand following alcohol cue exposure in adult heavy drinkers

Alcohol use disorder is associated with overvaluation of alcohol relative to other rewards, in part due to dynamic increases in value in response to alcohol-related cues. In a neuroeconomic framework, alcohol cues increase behavioral economic demand for alcohol, but the neural correlates these cue effects are unknown. This functional magnetic resonance imaging study combined a neuroeconomic alcohol purchase task with an alcohol cue exposure in 72 heavy drinkers with established sensitivity to alcohol cues (51 % female; mean age=33.74). Participants reported how many drinks they would consume from $0-$80/drink following exposure to neutral and alcohol images in a fixed order. Participants purchased significantly more drinks in the alcohol compared to the neutral condition, which was also evident for demand indices (i.e., intensity, breakpoint, Omax, elasticity; ps<0.001; ds=0.46–0.92). Alcohol purchase decisions were associated with activation in rostral middle and medial frontal gyri, anterior insula, posterior parietal cortex, and dorsal striatum, among other regions. Activation was lower across regions in the alcohol relative to neutral cue condition, potentially due to greater automaticity of choices in the presence of alcohol cues or attenuation of responses due to fixed cue order. These results contribute to growing literature using neuroeconomics to understand alcohol misuse and provide a foundation for future research investigating decision-making effects of environmental alcohol triggers.

Abstract 347: Tumor necrosis factor related apoptosis inducing ligand (TRAIL)-induced cytokine production in TNBC promotes neutrophil chemotaxis and immune suppression

Abstract Background: TRAIL induces apoptosis in many preclinical cancer models including breast cancers and has been extensively studied as a potential cancer therapeutic. However, its efficacy in clinical trials is limited, suggesting an unknown modulatory mechanism responsible for lack of TRAIL activity in vivo. Here, we describe that TRAIL treatment elicits transcriptional changes in TNBC cells that alter the immune milieu. Method: We performed RNAseq of MDA-MB-231 cells treated with TRAIL for different time points, followed by validation with RT-PCR in various TNBC cells. RNAi, silencing key differentially regulated genes, along with RT-PCR, ELISA and CHIP assays, were used to validate RNAseq findings. Elucidation of the functional relevance of the outcome was supported both in vitro and in vivo by chemotaxis assay, cytotoxicity assay, RNAseq analysis of donor isolated neutrophils and intravital microscopy, CODEX analysis in TNBC xenografts from mice treated with the TRAIL respectively. Results: TRAIL treatment of the TNBC significantly induced expression of several cytokines, such as CXCLs 1, 2, 3, 8,11 and IL6, both in vitro and in vivo which are known to affect neutrophil function. Mechanistically, induction of these cytokines was predominantly mediated by death receptor 5 and caspase-8 protein, but not caspase-8 enzymatic activity. GSEA of the RNAseq data indicated that NFKB pathway was significantly enriched. Concordantly, we confirmed that both canonical NFKB1 and non-canonical NFKB2 pathways were activated by TRAIL in vitro and in vivo. However, the induction of the cytokine mRNAs was primarily dependent on the NFKB2 pathway. Neutrophils isolated from healthy human donors incubated with supernatants from TNBC cells in vitro indicated that TRAIL-induced CXCLs and IL6 significantly increased neutrophil chemotaxis. Additionally, CODEX analysis as well as intravital imaging confirmed that TRAIL treatment increases the number of neutrophils in the tumor. Preincubation of neutrophils with supernatants from TRAIL-treated TNBC significantly inhibited their cytotoxic effect against TNBCs. Further, transcriptome analysis of neutrophils incubated with either TRAIL or supernatant of TRAIL treated TNBC revealed significant enrichment of expression of inflammatory cytokine genes, immune modulating and immune checkpoint genes like PDL1. Functional studies with these neutrophils confirmed their suppressive effect on T cell function as well as the effect of TRAIL on decreased neutrophil apoptosis. Conclusion: Collectively, our study suggests the novel role of TRAIL-induced NFKB2-dependent cytokine production promoting neutrophil chemotaxis and immune suppression. This study implies that alterations in the innate immune system may modulate the effects of TRAIL on TNBC tumors. Citation Format: Manjari Kundu, Yoshimi Endo Greer, Lisa Ann Ridnour, David A Wink, Yeap S. Ng, Roberto Weigert, Stan Lipkowitz. Tumor necrosis factor related apoptosis inducing ligand (TRAIL)-induced cytokine production in TNBC promotes neutrophil chemotaxis and immune suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 347.

Abstract 6139: Genetic modifiers of KRAS-mutant colorectal cancer

Abstract Somatic driver mutations in KRAS are found in ~40% of colorectal cancers (CRCs) and are associated with worse outcomes. In the US, individuals of African ancestry with CRC are more likely to have tumors with KRAS mutations than individuals of European ancestry, even after considering differences in socioeconomic status and other risk factors. Germline variants can influence somatic mutation events and may provide a selective advantage such that a mutated cell is more likely to progress to a cancer. Germline variants have been found to associate with somatic mutations in a variety of tumors. Based on ancestry differences in KRAS tumor mutation frequency, we hypothesized that germline variants altering molecular pathways that support KRAS tumorigenesis will associate with KRAS-mutant status. To test this, we performed a two-step association study using KRAS mutation status as our phenotype in individuals with microsatellite stable CRC. First, we conducted a discovery analysis using genome-wide germline genotypes and somatic mutation data from 6,386 non-Hispanic White CRC patients from the Cancer Genome Atlas and the GECCO consortium, considering sex and tumor location as covariates. To select variants for additional study, we used in-silico tools, such as expression quantitative trait loci (eQTL), to predict how associated variants may alter gene expression and chromatin. After reviewing relevant genomic features, excluding SNVs with minor allele frequency &amp;lt;10% and pruning SNVs in linkage disequilibrium (r2&amp;gt;0.75), we identified candidate loci for a multi-ancestry independent validation analysis. In the discovery analysis, no variants met genome-wide significance thresholds (p-value &amp;lt;5x10-8), though 3 SNVs associated with KRAS mutation status with p-values &amp;lt;1x10-6 and 50 with p-values &amp;lt;1x10-5. Of these 50 SNVs, 21 met criteria for validation. We chose 80 additional SNVs (p-value &amp;lt;1x10-4) with predicted functional effect, for a total of 101 SNVs for analyses. In our preliminary validation analysis of 1,487 individuals grouped by self-reported race, no variants had a significant association in all populations. The rs726800 variant showed nominal association with KRAS mutation status (p-value=0.028) in self-reported non-Hispanic White individuals. This variant is within potential cis-regulatory distance of HS3ST3A1 and COX10-DT, both of which have been reported as cancer biomarkers. Nominal associations were identified in self-reported Black (n=7 SNVs) and Hispanic or Latino (n=3 SNVs) individuals, though both groups had limited sample sizes. Additional validation samples are being analyzed, as well as analysis with adjustment for genetic ancestry. Future functional studies of top candidate variants will add mechanistic insight. In summary, we identified germline SNVs that associate with KRAS somatic mutations in CRC and may inform the cellular context that supports development of colorectal tumors with KRAS mutations. Citation Format: Nijole P. Tjader, Johnny Ramroop, Tanish Gandhi, Peter T. Campbell, Andrew T. Chan, Steven Gallinger, Graham G. Giles, Marc J. Gunter, Tabitha A. Harrison, Michael Hoffmeister, Polly A. Newcomb, Shuji Ogino, Amanda I. Phipps, Conghui Qu, Robert E. Schoen, Andrew Pellatt, Michael O. Woods, Bethany Van Guelpen, Patrick Stevens, Heather Hampel, Ulrike Peters, Joseph P. McElroy, Amanda E. Toland. Genetic modifiers of KRAS-mutant colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6139.

Characterizing the Visual Cortex Function in Cognitive Task-Induced Mental Load: An Functional Magnetic Resonance Imaging Study.

Introduction: The mental load caused by simultaneous multitasking can affect visual information processing and reduce its ability. This study investigated the effect of mental load caused by cognitive tasks simultaneously with visual task on the number of active voxels in the visual cortex. Methods: This study recruited 22 individuals with a mean age of 24.72 ± 5.47 years. 3-Tesla functional magnetic resonance imaging (fMRI) was used to examine the functions of the visual cortex and amygdala region during three different task conditions: visual task alone, visual task with an auditory n-back task, and visual task with an arithmetic task. The visual stimuli consisted of Gabor patches with a contrast of 55% at spatial frequencies of 0.25, 4, and 9 cycles per degree (cpd). These were presented in three trials of eight blocks with a stimulation time of 12 sec and a rest time of 14 sec. Results: Activated brain voxels in the primary, secondary, and associated visual cortex areas were reduced in response to the mental load imposed by the n-back and arithmetic tasks. This reduction was greater for a spatial frequency of 0.25 cpd in the n-back task condition and spatial frequency of 9 cpd in the arithmetic task condition. In addition, the amygdala was stimulated in 2-back task and arithmetic task conditions. Conclusions: This study revealed a decline in the number of activated voxels of the visual cortex due to the mental load caused by simultaneous cognitive tasks, confirming the findings of previous psychophysical studies.

Density functional theory study of Fe(CO)5 and [Fe(CO)6]2+ complexes catalyzed the water gas shift reaction extended to the aqueous phase

The WGSR with Fe(CO)5 and [Fe(CO)6]2+ complexes were investigated using DFT in both gaseous and aqueous phases. Thermodynamic analyses indicated that the high dielectric constant of water inhibits the activity of OH− and [Fe(CO)6]2+ ions, leading to a substantial decrease in the energy released during the nucleophilic attack of OH− on Fe(CO)5 and [Fe(CO)6]2+ in the aqueous phase. In addition, a potential low energy pathway has been discovered by designing new transition states for the final step of Fe(CO)5-catalyzed WGSR: 1 → 6 → 2 → 3 → 4 → 5(a) → 5(b) → 6. According to the results of the ESM, [Fe(CO)6]2+ possesses the highest gaseous phase TOF value (1.27×10−18 s–1), both Fe(CO)5 and [Fe(CO)6]2+ have higher TOF values in the gaseous phase compared to the aqueous phase, which increases with rising temperature. Therefore, these results suggest that [Fe(CO)6]2+ serves as the advantageous catalyst involved WGSR in the aqueous phase.

Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40

BackgroundCryptosporidium parvum is an apicomplexan zoonotic parasite causing the diarrheal illness cryptosporidiosis in humans and animals. To invade the host intestinal epithelial cells, parasitic proteins expressed on the surface of sporozoites interact with host cells to facilitate the formation of parasitophorous vacuole for the parasite to reside and develop. The gp40 of C. parvum, named Cpgp40 and located on the surface of sporozoites, was proven to participate in the process of host cell invasion.MethodsWe utilized the purified Cpgp40 as a bait to obtain host cell proteins interacting with Cpgp40 through the glutathione S-transferase (GST) pull-down method. In vitro analysis, through bimolecular fluorescence complementation assay (BiFC) and coimmunoprecipitation (Co-IP), confirmed the solid interaction between Cpgp40 and ENO1. In addition, by using protein mutation and parasite infection rate analysis, it was demonstrated that ENO1 plays an important role in the C. parvum invasion of HCT-8 cells.ResultsTo illustrate the functional activity of Cpgp40 interacting with host cells, we identified the alpha-enolase protein (ENO1) from HCT-8 cells, which showed direct interaction with Cpgp40. The mRNA level of ENO1 gene was significantly decreased at 3 and 24 h after C. parvum infection. Antibodies and siRNA specific to ENO1 showed the ability to neutralize C. parvum infection in vitro, which indicated the participation of ENO1 during the parasite invasion of HCT-8 cells. In addition, we further demonstrated that ENO1 protein was involved in the regulation of cytoplasmic matrix of HCT-8 cells during C. parvum invasion. Functional study of the protein mutation illustrated that ENO1 was also required for the endogenous development of C. parvum.ConclusionsIn this study, we utilized the purified Cpgp40 as a bait to obtain host cell proteins ENO1 interacting with Cpgp40. Functional studies illustrated that the host cell protein ENO1 was involved in the regulation of tight junction and adherent junction proteins during C. parvum invasion and was required for endogenous development of C. parvum.Graphical