bluuebunny/crossref_metadata_2025_split huggingface dataset:
The study of circulating biomarker panels in early diagnosis of Alzheimer's disease
Authors: Zhou Wen‐xia , Jiang Ning , Ruan Can‐jun , Cheng Xiao‐rui , Zhang Yong‐xiang 
Published: November 2023

Early, reliable and non‐invasive diagnosis of Alzheimer disease (AD) is a great challenge, which makes the prognosis and therapeutic interventions quite difficult. We aimed to investigate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a which were screened from 877 microRNAs in 60 serum samples with a microarray platform in prior study in serum of cognitively normal controls (CNC), mild cognitive impairment (MCI), a potential preliminary stage of AD, and AD and identify microRNA panel for predicting and diagnosing AD. Quantitative reverse‐transcriptase polymerase chain reaction assay was applied to evaluate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a with two independent cohorts including 202 participants CNC, MCI and AD. Logistic regression model based on microRNA panel was constructed using a training cohort (n=150) and then validated using an independent cohort (n=52). Area under the receiver operating characteristic curve (AUC) was used to evaluate diagnostic accuracy. The expressions of the four microRNAs were significantly decreased in MCI and AD versus CNC, and positively correlated with mini mental state examination (MMSE) score. Then, we identified a microRNA panel with the four microRNAs that demonstrated good diagnostic performance for MCI (AUC=0.788 and 0.815 for training and validation data set, respectively) and AD (AUC=0.769 and 0.823 for training and validation data set, respectively). When combined with MMSE score, the diagnostic performance of the microRNA panel was further improved. These results suggested that the serum microRNA panel we found has considerable clinical value in diagnosing AD, and the four microRNAs are potential circulating biomarkers.Support or Funding InformationThe project supported by the National Science and Technology Ministry (2011ZX09102‐101‐07, 2012ZX09301003‐002‐001, 2012BAI29B07)
Rapamycin as a Novel Therapeutic for Alzheimer's Disease: Prevention Assessed Through Neuroimaging
Authors: Wang Amy C , Ma David Wang , Parikh Ishita , Bakshi Vikas , Hoffman Jared , Thalman Scott , Lin Ai‐Ling 
Published: November 2023

Alzheimer's disease (AD) is the sixth leading cause of death in the United States. Although AD research is bustling, a concrete and effective treatment has yet to be discovered. This study aimed to assess vascular magnetic resonance imaging (MRI) as an early detection method for AD, and to then test the potential of FDA‐approved drug Rapamycin (Rapa) in treating pre‐symptomatic AD. Rapa is capable of increasing lifespan in mammals and inhibiting unwanted cell mobility, such as cancer cell growth. This study is the first to connect Rapa's capabilities with AD. In order to analyze Rapa's efficacy, neuroimaging and behavioral tests were conducted on mice that carry the apolipoprotein E4 gene (APOE4), the strongest genetic risk factor for AD. Methodology included MRI, magnetic resonance spectroscopy (MRS), Blood‐Brain Barrier (BBB) integrity and neuroinflammation determinations, Radial Arm Water Maze (RAWM), and Novel Object Recognition Test (NOR). These methods provided a comprehensive analysis of the subjects' neurological health and cognitive functions by giving insight into cerebral blood flow, brain metabolites, spatial memory, recognition memory, and neuroinflammation. Vascular MRI effectively pinpointed vascular defects and therefore accurately identified early AD. Data strongly supported the initial hypothesis that with the introduction of Rapa, APOE4 mice will show improved cognition, memory, and neurological health. Cerebral blood flow and crucial brain metabolites were restored in Rapa mice; Rapa mice exhibited cogent memory in comparison to their counterparts in cognitive tests; Rapa reduced neuroinflammation‐‐all with significance. Thus, vascular neuroimaging is a promising early AD detection technique and Rapa holds high potential in preventing AD development.Support or Funding InformationThe work was supported by funding from the National Institute on Aging (NIA) of NIH (K01AG040164 to A‐LL) and NIH CTSA at the University of Kentucky (UL1TR0000117; pilot grant to A‐LL).
Zfra restores memory deficits in Alzheimer's disease triple transgenic mice by blocking tau aggregation and amyloid beta formation
Authors: Lin Yu‐Hao , Shih Yao‐Hsiang , Lee Ming‐Hui , Lin Sing‐Ru , Chang Jean‐Yun , Sze Chun‐I , Kuo Yu‐Min , Chang Nan‐Shan 
Published: November 2023

IntroductionZfra is a naturally occurring 31‐amino‐acid protein. Synthetic peptides Zfra1–31 and Zfra4–10 are known to effectively block the growth of many types of cancer cells.Methods10‐month‐old triple‐transgenic (3×Tg) mice for Alzheimer's disease (AD) received synthetic Zfra peptides via tail vein injections, followed by examining restoration of memory deficits.ResultsZfra significantly down‐regulated tau and amyloid beta (Abeta) aggregates in the brains of 3×Tg mice, and effectively restored their memory capabilities. Zfra inhibited melanoma‐induced neuronal death in the hippocampus and plaque formation in the cortex. Mechanistically, Zfra blocked the aggregation of Abeta42 and many serine‐containing peptides in vitro, and bound cytosolic proteins for accelerating their degradation in ubiquitin/proteasome‐independent manner.DiscussionZfra peptides exhibit a strong efficacy in blocking tau aggregation and Abeta formation and restores memory deficits in 3×Tg mice, suggesting its potential for treatment of AD.Support or Funding InformationDepartment of Defense USA; Ministry of Science and Technology, Taiwan; National Cheng Kung University Top Notch Project
Co‐ultra PEALut Enhances Neuronal Recovery After a Moderate Traumatic Brain Injury
Authors: Cuzzocrea Salvatore , Paterniti Irene , Campolo Michela , Crupi Rosalia , Filippone Alessia , Cordaro Marika , Bruschetta Giuseppe , Esposito Emanuela 
Published: November 2023

Traumatic brain injury (TBI) is a major health problem worldwide. Currently, there is no effective treatment to improve neural structural repair and functional recovery. Recently, has been demonstrated that hippocampal injury‐induced neurogenesis is one mechanism underlying endogenous repair following TBI. The aim of this study was to evaluate the neuro‐regenerative properties co‐ultra PEALut (constituted by the association of palmitoylethanolamide (PEA), with the flavonoid luteolin (Lut)) in a mouse model of TBI until 7 days. TBI was induced in mice by the controlled cortical impact (CCI), one of the most common models of TBI; the mice were orally administered co‐ultra PEALut (1mg/Kg) 1h after trauma and daily for 72h and 7d. A conspicuous neurogenesis was seen in mice 72h and 7d after trauma. Co‐ultra PEALut treatment stimulated neuronal reconstitution process restoring the basal level of new neurons (by BrdU staining) and mature neurons (by doublecortin staining). Moreover this positive effect was associated with an important upregulation of neurotrophic factors (BDNF and NT‐3), ultimately leading to improvement in memory recall on behavioral testing (Morris water maze test).Co‐ultra PEALut could represent a therapeutic potential of augmenting the endogenous repair response for treating TBI.
Characterization of Rap1A‐dependent activation of PLCɛ
Authors: Sieng Monita , Garland‐Kuntz Elisabeth E. , Lyon Angeline M. 
Published: November 2023

Phospholipase C (PLC) enzymes hydrolyze the lipid phosphatidylinositol 4,5‐bisphosphate (PIP2) to produce diacylglycerol (DAG) and inositol 1,4,5‐triphosphate (IP3) which activate protein kinase C and promote intracellular Ca2+ release, respectively. These second messengers are critical for many physiological processes, especially within the cardiovascular system where they regulate cell growth and survival. The PLCɛ subfamily has emerged as a key player in cardiovascular health, due to its involvement in cardiac contractility and regulation of hypertrophic gene expression. PLCɛ is activated downstream of G protein coupled receptors, including the β adrenergic receptors, through direct interactions with small G proteins. The best characterized PLCɛ activator is the small G protein Rap1A. In contrast to canonical PLC activation, Rap1A· GTP binds to PLCɛ and translocates the activation complex to the perinuclear membrane, where phosphatidylinositol‐4‐phosphate (PI4P) is the preferred substrate. Activation by Rap1A is associated with the onset of cardiac hypertrophy. However, while Rap1A is known to interact with the C‐terminal Ras association (RA) domain of PLCɛ, little is known about the molecular basis of activation. We seek to characterize the interactions between Rap1A and PLCɛ using structural and functional studies to map the entire Rap1A binding site on PLCɛ, and determine whether activation results in conformational changes that release autoinhibition and/or increase membrane association. These studies provide the first molecular details of the Rap1A‐dependent activation of PLCɛ.Support or Funding InformationFunding provided by American Heart Association Grant 16SDG29920017 to Angeline M. Lyon.
Structure‐Based Design of IKKɛ and TBK1 Inhibitors for the Treatment of Obesity
Authors: Beyett Tyler S , Labudde Emily , Tesmer John J. G. 
Published: November 2023

Obesity is a leading factor in the development of type 2 diabetes. It has been shown that inhibitor of NF‐κB kinase epsilon (IKKɛ) and TANK‐binding kinase 1 (TBK1) are upregulated in adipocytes of obese individuals. Knockdown of either kinase results in significant weight loss and increased insulin sensitivity. High‐throughput screening identified the FDA‐approved drug amlexanox as an ATP‐competitive inhibitor of both kinases which produces significant weight loss when administered to obese mice. Amlexanox has poor solubility and metabolic profile when administered orally, necessitating the design of improved analogs for clinical use. Docking simulations have not provided consistent binding modes for amlexanox due to the planar and somewhat symmetrical structure of the molecule, limiting rational design of improved analogs.To better understand the molecular mechanism of inhibition and to guide further development, we sought to obtain co‐crystals structures of IKKɛ and TBK1 with amlexanox and related analogs. While crystallization of IKKɛ has been unsuccessful, we generated two crystal forms of TBK1 in complex with the potent inhibitor MRT67307 but were unable to obtain co‐crystals in the presence of amlexanox. Soaking these crystals in amlexanox resulted in a significant decrease in diffraction quality and partial density for the original inhibitor. TBK1·amlexanox crystals of a moderate resolution (3.7 Å) were obtained via microseeding and subsequent macroseeding. The resulting structure shows amlexanox binding along the hinge of the kinase and suggests that the carboxylate on amlexanox may form a salt bridge with Lys38 and/or Thr156. Differential scanning fluorometry with an K38A mutant shows a loss in thermal stabilization by amlexanox, and analogs with modifications to the carboxylate display decreased or abolished ability to inhibit TBK1. Knowledge of the potential importance of the carboxylate‐Lys38 interaction will guide the development of future amlexanox analogs. Presently, we have developed amlexanox analogs with improved efficacy in mouse models and are progressing them towards the clinic.Support or Funding InformationNational Institutes of Health, Pharmacological Sciences Training Program (T32‐GM007767), Rackham Graduate School at the University of Michigan
A Quantitative Morphometric Analysis of the Effects of Sub‐Chronic Cadmium Exposure on the Proximal Tubule of the Rat
Authors: Hublikar Ishaan , Edwards Joshua R , Lamar Peter C , Gasiorowski Joshua Z , Prozialeck Walter C 
Published: November 2023

Cadmium (Cd) is a nephrotoxic environmental pollutant that causes insidious injury to the proximal tubule that results in polyuria and proteinuria. Previous studies from our laboratory have shown that the early stages of Cd‐induced kidney injury may involve disruption of the adhering junctions between epithelial cells of the proximal tubule. In the present study, we utilized the Image J computer program to quantify the effects of sub‐chronic Cd exposure on the morphology of the rat proximal tubule. Adult male Sprague‐Dawley rats (n=6) were treated with CdCl2 (0.6 mg/kg, in saline vehicle) by subcutaneous injection, 5 days per week for 12 weeks, while control animals (n=6) received saline vehicle alone. At the end of the treatment period, 24 hour urine samples were collected and assayed for levels of protein, cystatin C, beta‐2 microglobulin and kidney injury molecule 1 (Kim 1). In addition, the animals were euthanized and the kidneys were removed, fixed in formalin and processed for histopathologic analysis. Hematoxylin/eosin‐stained sections (4 microns thick) were viewed using a 40× objective lens, and digital images (n=15 control and 15 Cd‐treated) of the renal cortex were captured. The Image J computer program was used to identify, by hand drawing, cross‐sections of proximal tubules (n=5–8 per image). In addition, Image J was used to define individual components of the cross‐sectional images of proximal tubules. Specific components that were defined included: total cross sectional area, luminal cross sectional area, area of gaps between cells and total cellular area. The results showed that Cd caused an increase in urine volume and urinary levels of protein, Kim 1, beta‐2 microglobulin and cystatin C, with no change in creatinine excretion. The morphometric analyses showed that Cd had no significant effect on total cross sectional tubular area or cellular area. However, it caused a slight decrease (36%, p<0.01) in luminal cross sectional area and a marked increase (300%, p<0.001) in gap areas between cells. Additional studies indicated that at the time these changes were occurring, there was no evidence of necrosis and only a slight increase in the number of apoptotic cells in the proximal tubule. These results are consistent with previous findings suggesting that the early stages of Cd‐nephrotoxicity involve the disruption of epithelial cell‐cell junctions in the proximal tubule.Support or Funding InformationSupported by restricted funds from the Department of Pharmacology, Midwestern University.
Neurotransmitter Systems in Mouse Airways
Authors: Kieffer Cameron M , Tu Yaping , Abel Peter W 
Published: November 2023

Autonomic and sensory nerves containing a variety of neurotransmitters innervate mouse lungs and control airway diameter. The objective of this study was to characterize the neurotransmitter systems that cause constriction of murine airways. Precision cut lung slices (PCLS) from CF‐1 mice were exposed to electric field stimulation (EFS) using parameters known to activate neurotransmitter release (1.0 ms, 50 V, 1–30 Hz). To measure constriction, changes in airway luminal area in response to EFS were captured as video images and quantified using ImageJ. EFS caused airway constriction in a frequency‐dependent manner that was best fit by a biphasic curve. PCLS taken from human lungs showed a similar biphasic fit. Neuron‐specific stimulation was verified by Mg2+ blockade. Maximum airway constriction to 30 Hz EFS in murine PCLS was 51.8 ± 3.0%. Interestingly, the muscarinic receptor antagonist atropine (10 μM) blocked 99.5 ± 7.2% of EFS induced constriction at 1 Hz, but only blocked 23.3 ± 3.8% of EFS induced constriction at 30 Hz and eliminated the first phase but not the second phase of the biphasic EFS response. These data are consistent with parasympathetic constriction of airways by acetylcholine at lower EFS frequencies. Treatment with capsaicin to deplete sensory neurotransmitters also altered the EFS response supporting the presence of sensory neurotransmitter systems. Taken together these data provide evidence for a robust, multi‐component innervation of mouse lungs. Elucidating the neurotransmitter systems causing nerve mediated constriction and dilatation of airways may identify novel therapeutic targets for treatment of asthma, COPD, and other lung diseases.Support or Funding InformationState of Nebraska LB595
Phosphatidylinositol‐3,4,5‐trisphosphate‐dependent Rac exchange factor 1 (P‐Rex1) Exacerbates Asthmatic Airway Smooth Muscle Cell Proliferation via Rac1‐dependent Signal Pathways
Authors: Huang Yapei , Xie Yan , Jiang Haihong , Abel Peter W. , Panettieri Reynold A. , Casale Thomas B. , Tu Yaping 
Published: November 2023

Background and ObjectiveHuman airway smooth muscle (HASM) hyperplasia and remodeling are characteristic features of airways that contribute to their irreversible obstruction in patients with severe asthma. Accumulating evidence suggests that cell proliferation contributes to airway smooth muscle hyperplasia and remodeling. Although the role of HASM hyperplasia and remodeling in asthma is well established, the molecular mechanisms regulating the proliferation of HASM cells in asthma‐related conditions are poorly understood. P‐Rex1, a Rac‐selective guanine nucleotide exchange factor (GEF), plays important roles in both normal physiology and pathological conditions. The objective of our study was to define the pathological importance and mechanisms of upregulated P‐Rex1 in development of ASM hyperplasia and remodeling of severe asthma.MethodsWe used immunohistochemistry to compare P‐Rex1 protein expression in lung tissues between human asthmatic and control subjects. Western blot was used to determine changes in P‐Rex1 expression in primary HASM cells isolated from asthmatic and non‐asthmatic subjects. P‐Rex1 expression levels in HASM cells were manipulated by overexpression of recombinant P‐Rex1 or its specific siRNA. The proliferation rate of HASM cells was measured by 5‐Bromo‐2‐deoxyUridine (BrdU) assays.ResultsImmunohistochemistry staining revealed that P‐Rex1 was expressed in the epithelial layer of the bronchioles. Positive staining was also observed in HASM cells. However, samples from asthmatic subjects had a 4‐fold higher density of P‐Rex1 protein compared to those from control subjects (p < 0.01). Western blot assay also showed 3‐fold higher expression of P‐Rex1 protein in asthmatic HASM cells as compared to non‐asthmatic HASM cells (p<0.01). Overexpression of P‐Rex1 in non‐asthmatic HASM cells increased platelet‐derived growth factor (PDGF)‐stimulated cell proliferation by more than 2‐fold. Conversely, silencing P‐Rex1 in asthmatic HASM cells reduced PDGF‐stimulated cell proliferation by over 70%. Interestingly, treatment with Rac1 inhibitor NSC23766 not only significantly inhibited PDGF‐stimulated cell proliferation in 4 different asthmatic HASM cells but also largely abolished recombinant P‐Rex1 augment of proliferation in non‐asthmatic HASM cells.ConclusionIncrease of P‐Rex1 protein promotes HASM cell proliferation via Rac1‐dependent pathways, which in turn causes HASM hyperplasia and remodeling, leading to irreversible airway obstruction.Support or Funding InformationThis work was supported by grants from the National Institutes of Health (5P20GM103489 and R01HL116849) and Nebraska State LB595 research program
Characterization of UDP‐glycosyltransferase 3A (UGT3A) Variants in Tobacco Carcinogen Metabolism
Authors: Vergara Ana G , Watson Christy J , Lazarus Philip 
Published: November 2023

The UDP‐glycosyltransferase (UGT) family are phase II enzymes important in the metabolism of endogenous and exogenous substances including steroids and a variety of different carcinogens and drugs. The UGT3A subfamily consists of UGTs 3A1 and 3A2, which may play a role in the local detoxification of tobacco carcinogens. The objective of the present study is to identify UGT3A protein expression in human aerodigestive tract tissues, determine their glycosylation activity against tobacco carcinogens, and assess the potential functional role of UGT3A variants in tobacco‐related cancer susceptibility. As determined by Western blot analysis, UGT3A1 exhibited the highest expression in liver, followed by lung and several aerodigestive tract tissues. UGT3A2 exhibited the highest expression in floor of mouth, followed by trachea and other aerodigestive tract tissues. Of the tissues tested, liver exhibited the least UGT3A2 expression. Microsomes prepared from HEK293 cells overexpressing wild‐type UGT3A1 demonstrated glycosylation activity, as observed by reverse‐phase ultra‐pressure liquid chromatography, against polycyclic aromatic hydrocarbons (PAHs) including 1‐hydroxybenzo(a)pyrene (1‐OH‐B(a)P) and 8‐OH‐B(a)P. Microsomes prepared from HEK293 cells overexpressing wild‐type UGT3A2 exhibited significant glycosylation activity against a variety of PAHs including, 1‐OH‐B(a)P), 3‐OH‐B(a)P, 7‐OH‐B(a)P, 8‐OH‐B(a)P and 9‐OH‐B(a)P; additional PAHs and other tobacco carcinogens are currently being tested for both UGT3A enzymes. Preliminary kinetic analysis demonstrated a low Km, in the range of 130 – 330 μM, against all PAHs tested by UGT3A2, similar to that observed previously for UGT2A1 (Bushey RT, 2011). In addition, seven variants (>1% prevalence) for each UGT3A enzyme have been overexpressed in HEK293 cells and are currently being screened for altered activity against PAHs in vitro. These data suggest that UGT3A enzymes are important for the local detoxification of tobacco carcinogens, specifically PAHs. Inter‐individual differences in UGT3A expression and/or activity may result in variations in the detoxification of tobacco carcinogens and may play a role in tobacco‐related cancer risk.Support or Funding InformationThis study is supported by the National Institute of Environmental Health Sciences (NIEHS) of the National Institutes of Health to P Lazarus (Grant R01‐ES025460) and a NIEHS Research Supplement to Promote Diversity in Health‐Related Research to AG Vergara (Grant R01‐ES025460‐02S1).
Human Fetal Sulfotransferase 1C4: Unique Properties and Novel Substrates
Authors: Guidry Amber L. , Runge‐Morris Melissa , Falany Charles N. 
Published: November 2023

Sulfoconjugation of xenobiotics and endogenous compounds by the human cytosolic sulfotransferases (hSULTs) modifies the substrates' functions and increases the compounds' excretion from the body. The hSULT1C subfamily, in contrast with most of the other human SULTs, is primarily expressed during human fetal development; however, the biological role of these fetal isoforms remains uncharacterized. The purpose of this study was to generate pure hSULT1C4 and to characterize the enzyme's structure, activity, and physiological substrates. Based on hSULT1C4's native biochemical properties, a novel purification protocol utilizing affinity chromatography and ion exchange chromatography was established. The small phenolic compound 1‐naphthol was analyzed in combination with the obligate sulfonate donor PAPS (3′‐phosphoadenosine 5′‐phosphosulfate) to optimize hSULT1C4 activity assays. To begin characterizing hSULT1C4, the affinity of the enzyme for PAPS and the reaction product PAP (3′, 5′‐diphosphoadenosine) was determined by measuring the Michaelis constant of PAPS (7.0 ± 1.0 μM) and the dissociation constant of PAP (2.8 ± 0.2 μM) and PAPS (4.8 ± 0.8 μM). Human SULT1C4 has a reduced affinity for PAPS compared to PAP. Previous studies have shown hSULT1C4 sulfonates catechol estrogens; therefore, endogenous and environmental estrogenic compounds were screened as substrates for sulfonation by hSULT1C4. The hSULT1C4 activity assays revealed that all of the screened estrogenic compounds (genistein, daidzein, apigenin, chrysin, 6,4′‐dihydroxyflavone, estradiol, and bisphenol A) are substrates for hSULT1C4. Based on the enzyme's expression in the human fetus and its affinity for sulfonating estrogenic compounds, hSULT1C4's physiological function may be to metabolize various dietary compounds and hormonal signaling molecules during fetal development.Support or Funding InformationThis work was supported by the National Institutes of Health Grant GM038953 and National Institutes of Health Grant ES022606.