Submerged fermentation of lentil protein isolate and its impact on protein functionality, nutrition, and volatile profiles.

Fermentation of pulses as a clean processing technique has been reported to have a favorable impact on the functional and nutritional quality of the starting materials. Compared to commonly fermented pulses such as peas and chickpeas, limited information is available on the effect of fermentation on lentils, especially when using a high protein isolate (>80% protein) as compared to seeds or flours. Therefore, in the present work, lentil protein isolate was used as a feedstock for submerged fermentation with Aspergillus niger, Aspergillus oryzae, or Lactobacillus plantarum. After 48 h, the samples showed increased protein content with enhanced solubility and oil-holding capacity. Controlled fermentation, as opposed to spontaneous fermentation, maintained the high foaming capacity; however, all fermented samples had lower foam and emulsion stabilizing properties and reduced water-holding capacity compared to the control. The fermented proteins were also less digestible, possibly due to an increase in phenolics and saponins. New volatile compounds were identified in fermented samples that show promise for improved sensory attributes. Significant differences were observed in specific quality attributes depending on the microbial strain used. Further research is required to better understand the fermentative metabolism of microbial communities when provided high-protein lentil ingredients as growth substrates. PRACTICAL APPLICATION: Fermented lentil protein isolate has promising flavor profiles that may improve its sensory properties for food application.

Noninvasive Monitoring of Early Cardiac Fibrosis in Diabetic Mice by [68Ga]Ga-DOTA-FAPI-04 PET/CT Imaging.

Cardiac fibrosis represents one of the representative pathological characteristics in the diabetic heart. Active fibroblasts play an essential role in the progression of cardiac fibrosis. The technologies for noninvasive monitoring of activated fibroblasts still have to be investigated. The purpose of this study was to evaluate the feasibility of targeted fibroblast activation protein (FAP) molecular imaging in the early evaluation of diabetic cardiac fibrosis using [68Ga]Ga-DOTA-FAPI-04 PET/CT. PET/CT imaging was conducted in db/db mice and db/m mice at weeks 12 and 24. Diabetic heart injury was determined using echocardiography and serum biomarkers. Additionally, the levels of cardiac fibrosis were also assessed. In our study, conventional diagnostic modalities, including echocardiography and serum biomarkers, failed to monitor early-stage cardiac dysfunction and fibrosis in diabetic mice. Conversely, the results of [68Ga]Ga-DOTA-FAPI-04 PET/CT imaging demonstrated that diabetic mice had increased myocardial uptake of radioactive tracers in both early-stage and late-stage diabetes, consistent with the elevated FAP expression and increased cardiac fibrosis level. Notably, cardiac PET signals exhibited significant correlations with left ventricular ejection fractions, the E/A ratio, and the level of serum TGF-ß1, PIIINP, and sST2. The results demonstrated the potential of [68Ga]Ga-DOTA-FAPI-04 PET/CT imaging for visualizing activated fibroblasts and detecting early-stage diabetic heart injury and fibrosis noninvasively. They also demonstrated the clinical superiority of [68Ga]Ga-DOTA-FAPI-04 PET/CT imaging over echocardiography and serum biomarkers in the early monitoring of diabetes-related cardiac dysfunction and fibrosis.

Pathophysiology characterization of Alzheimer's disease in South China's aging population: for the Greater-Bay-Area Healthy Aging Brain Study (GHABS).

INTRODUCTION: The Guangdong-Hong Kong-Macao Greater-Bay-Area of South China has an 86 million population and faces a significant challenge of Alzheimer's disease (AD). However, the characteristics and prevalence of AD in this area are still unclear due to the rarely available community-based neuroimaging AD cohort. METHODS: Following the standard protocols of the Alzheimer's Disease Neuroimaging Initiative, the Greater-Bay-Area Healthy Aging Brain Study (GHABS) was initiated in 2021. GHABS participants completed clinical assessments, plasma biomarkers, genotyping, magnetic resonance imaging (MRI), ß-amyloid (Aß) positron emission tomography (PET) imaging, and tau PET imaging. The GHABS cohort focuses on pathophysiology characterization and early AD detection in the Guangdong-Hong Kong-Macao Greater Bay Area. In this study, we analyzed plasma Aß42/Aß40 (A), p-Tau181 (T), neurofilament light, and GFAP by Simoa in 470 Chinese older adults, and 301, 195, and 70 had MRI, Aß PET, and tau PET, respectively. Plasma biomarkers, Aß PET, tau PET, hippocampal volume, and temporal-metaROI cortical thickness were compared between normal control (NC), subjective cognitive decline (SCD), mild cognitive impairment (MCI), and dementia groups, controlling for age, sex, and APOE-ε4. The prevalence of plasma A/T profiles and Aß PET positivity were also determined in different diagnostic groups. RESULTS: The aims, study design, data collection, and potential applications of GHABS are summarized. SCD individuals had significantly higher plasma p-Tau181 and plasma GFAP than the NC individuals. MCI and dementia patients showed more abnormal changes in all the plasma and neuroimaging biomarkers than NC and SCD individuals. The frequencies of plasma A+/T+ (NC; 5.9%, SCD: 8.2%, MCI: 25.3%, dementia: 64.9%) and Aß PET positivity (NC: 25.6%, SCD: 22.5%, MCI: 47.7%, dementia: 89.3%) were reported. DISCUSSION: The GHABS cohort may provide helpful guidance toward designing standard AD community cohorts in South China. This study, for the first time, reported the pathophysiology characterization of plasma biomarkers, Aß PET, tau PET, hippocampal atrophy, and AD-signature cortical thinning, as well as the prevalence of Aß PET positivity in the Guangdong-Hong Kong-Macao Greater Bay Area of China. These findings provide novel insights into understanding the characteristics of abnormal AD pathological changes in South China's older population.

Synthesis, Screening, and Evaluation of Theranostic Molecular CPCR4-Based Probe Targeting CXCR4.

Chemokines and chemokine receptors are indispensable to play a key role in the development of malignant tumors. As one of the most widely expressed chemokine receptors, chemokine (C-X-C motif) receptor 4 (CXCR4) has been a popular research focus. In most tumors, CXCR4 expression is significantly upregulated. Moreover, integrated nuclide diagnosis and therapy targeting CXCR4 show great potential. [68Ga]Ga-pentixafor, a radioligand targeting CXCR4, exhibits a strong affinity for CXCR4 both in vivo and in vitro. However, [177Lu]Lu-pentixather, the therapeutic companion of [68Ga]Ga-pentixafor, requires significant refinement to mitigate its pronounced hepatic biodistribution. The objective of this study was to synthesize theranostic molecular tracers with superior CXCR4 targeting functions. The Daudi cell line, which highly expressed CXCR4, and the MM.1S cell line, which weakly expressed CXCR4, were used in this study. Based on the pharmacophore cyclo (-d-Tyr-n-me-d-Orn-l-Arg-L-2-NAL-Gly-) (CPCR4) of pentixafor, six tracers were synthesized: [124I]I-1 ([124I]I-CPCR4), [99mTc]Tc-2 ([99mTc]Tc-HYNIC-CPCR4), [124I]I-3 ([124I]I-pentixafor), [18F]AlF-4 ([18F]AlF-NETA-CPCR4), [99mTc]Tc-5 ([99mTc]Tc-MAG3-CPCR4) and [124I]I-6 ([124I]I-pentixafor-Ga) and their radiochemical purities were all higher than 95%. After positron emission tomography (PET)/single-photon emission computed tomography (SPECT) imaging, the [124I]I-6 group exhibited the best target-nontarget ratio. At the same time, comparing the [68Ga]Ga-pentixafor group with the [124I]I-6 group, we found that the [124I]I-6 group had a better target-nontarget ratio and lower uptake in nontarget organs. Therefore, compound 6 was selected for therapeutic radionuclide (131I) labeling, and the tumor-bearing animal models were treated with [131I]I-6. The volume of the tumor site was significantly reduced in the treatment group compared with the control group, and no significant side effects were found. [124I]I-6 and [131I]I-6 showed excellent affinity for targeting CXCR4, and they showed great potential for the integrated diagnosis and treatment of tumors with high CXCR4 expression.

Association of ß-Amyloid, Microglial Activation, Cortical Thickness, and Metabolism in Older Adults Without Dementia.

BACKGROUND AND OBJECTIVES: Plasma ß-amyloid42 (Aß42)/Aß40 levels have shown promise in identifying Aß-PET positive individuals. This study explored the concordance and discordance of plasma Aß42/Aß40 positivity (Plasma±) with CSF Aß42/Aß40 positivity (CSF±) and Aß-PET positivity (PET±) in older adults without dementia. Associations of Aß deposition, cortical thickness, glucose metabolism, and microglial activation were also investigated. METHODS: We selected participants without dementia who had concurrent plasma Aß42/Aß40 and Aß-PET scans from the Alzheimer's Disease Neuroimaging Initiative cohort. Participants were categorized into Plasma±/PET± based on thresholds of composite 18F-florbetapir (FBP) standardized uptake value ratio (SUVR) ≥1.11 and plasma Aß42/Aß40 ≤0.1218. Aß-PET-negative individuals were further divided into Plasma±/CSF± (CSF Aß42/Aß40 ≤0.138), and the concordance and discordance of Aß42/Aß40 in the plasma and CSF were investigated. Baseline and slopes of regional FBP SUVR were compared among Plasma±/PET± groups, and associations of regional FBP SUVR, FDG SUVR, cortical thickness, and CSF soluble Triggering Receptor Expressed on Myeloid Cell 2 (sTREM2) levels were analyzed. RESULTS: One hundred eighty participants (mean age 72.7 years, 51.4% female, 96 cognitively unimpaired, and 84 with mild cognitive impairment) were included. We found that the proportion of Plasma+/PET- individuals was 6.14 times higher (odds ratio (OR) = 6.143, 95% confidence interval (CI) 2.740-16.185, p < 0.001) than that of Plasma-/PET+ individuals, and Plasma+/CSF- individuals showed 8.5 times larger percentage (OR = 8.5, 95% CI: 3.031-32.974, p < 0.001) than Plasma-/CSF+ individuals in Aß-PET-negative individuals. Besides, Plasma+/PET- individuals exhibited faster (p < 0.05) Aß accumulation predominantly in bilateral banks of superior temporal sulcus (BANKSSTS) and supramarginal, and superior parietal cortices compared with Plasma-/PET- individuals, despite no difference in baseline FBP SUVRs. In Plasma+/PET+ individuals, higher CSF sTREM2 levels correlated with slower BANKSSTS Aß accumulation (standardized ß (ßstd) = -0.418, 95% CI -0.681 to -0.154, p = 0.002). Conversely, thicker cortical thickness and higher glucose metabolism in supramarginal and superior parietal cortices were associated with faster (p < 0.05) CSF sTREM2 increase in Plasma+/PET- individuals rather than in Plasma+/PET+ individuals. DISCUSSION: These findings suggest that plasma Aß42/Aß40 abnormalities may predate CSF Aß42/Aß40 and Aß-PET abnormalities. Higher sTREM2-related microglial activation is linked to thicker cortical thickness and higher metabolism in early amyloidosis stages but tends to mitigate Aß accumulation primarily at relatively advanced stages.

Increased Cyclic Adenosine Monophosphate Responsive Element is Closely Associated with the Pathogenesis of Drug-resistant Epilepsy.

BACKGROUND: Drug-resistant epilepsy (DRE) is a refractory neurological disorder. There is ample evidence that suggest that γ-aminobutyric acid-a (GABAA) receptors could be one of the mechanisms responsible for the development of drug resistance in epilepsy. It is also known that the cAMP response element binding protein (CREB) plays a possible key role in the transcriptional regulation of GABAA. OBJECTIVE: This study explores the role of CREB in the development of DRE and the effect of CREB on GABA-related receptors in DRE. METHODS: The CREB expression was increased or decreased in the hippocampus of normal rats by lentiviral transfection, who then underwent the lithium-pilocarpine-induced epilepsy model. Phenobarbital (PB) sodium and carbamazepine (CBZ) were used to select a drug-resistant epileptic model. The expression levels of GABAA receptor α1, ß2, and γ2 subunits and CREB protein were measured in the rat hippocampus by western blot and fluorescent quantitative PCR. RESULTS: The frequency and duration of seizures increased in the overexpression group compared to that in the control group. In addition, the severity, frequency, and duration of seizures decreased in the group with decreased expression. The hippocampus analysis of the expression levels of the CREB protein and CREB mRNA yielded similar findings. Altering the CREB protein expression in the rat hippocampus could negatively regulate the expression and transcript levels of GABAA receptors α1, ß2, and γ2, suggesting that CREB may serve as a potential target for the development of treatment protocols and drugs for epilepsy. CONCLUSION: Our study shows that enhanced CREB expression promotes the development of DRE and negatively regulates GABAA receptor levels and that the inhibition of CREB expression may reduce the incidence of DRE.

Preclinical Evaluation of 99mTc-MAG3-5-Fab Targeting TREM2 in Lung Cancer Mouse Models: A Comparison with 99mTc-MAG3-5-F(ab')2.

Triggering receptor expressed on myeloid cells-2 (TREM2), which is expressed on the surface of tumor-associated macrophages (TAMs), has been found to play a major role in the diagnosis and treatment of tumors. TREM2 expression is significantly upregulated in tumor tissues, and therefore, targeting TREM2 for tumor imaging may be of value. Previously, we performed TREM2 targeting imaging by using 68Ga-NOTA-COG1410 or a 124I-labeled monoclonal antibody (mAb) and F(ab')2 in mouse models of colon and gastric tumors. However, some of the shortcomings of these probes (i.e., the high uptake of 68Ga-NOTA-COG1410 in the liver, the difficulty of obtaining iodine-124, and the long half-life of iodine-124) have hindered their clinical use. Herein, we sought to synthesize novel molecular probes targeting TREM2 that are more conducive to clinical translation, eliminating the interference of isotope availability and in vivo probe biodistribution issues. Therefore, we established A549 cell lines with negative human TREM2 (hTREM2) expression (GFP tag; hTREM2- A549) or upregulated hTREM2 expression (GFP tag; hTREM2+ A549) using lentiviral transfection and confirmed these with Western blotting and immunocytochemistry. We then prepared a mouse anti-human TREM2 (5-mAb) by immunizing with the hTREM2 antigen. The antibody fragments 5-F(ab')2 and 5-Fab were prepared from 5-mAb, and 99mTc-MAG3-5-F(ab')2 and 99mTc-MAG3-5-Fab were then synthesized with excellent stability and specificity. 99mTc-MAG3-5-F(ab')2 had a slightly higher in vitro affinity than 99mTc-MAG3-5-Fab (Kd = 3.32 ± 0.05 nmol versus 4.62 ± 0.85 nmol). 99mTc-MAG3-5-F(ab')2 and 99mTc-MAG3-5-Fab both showed excellent specificity: after adding a 100-fold precursor, the two probes binding to the cells were almost blocked. In vivo pharmacokinetics showed that the distribution and elimination half-lives of 99mTc-MAG3-5-Fab (T1/2α = 1.25 ± 0.30 min and T1/2ß = 21.98 ± 2.80 min, respectively) were significantly reduced compared to those of 99mTc-MAG3-5-F(ab')2 (T1/2α = 2.64 ± 0.37 min and T1/2ß = 86.55 ± 26.86 min, respectively). In micro single-photon emission computed tomography/computed tomography (micro-SPECT/CT) imaging, the tumor was clearly displayed at 1 h after 99mTc-MAG3-5-Fab injection, while the blood background was extremely low at 3 h, and the probe was mainly excreted through the kidneys and biliary tract. 99mTc-MAG3-5-F(ab')2 uptake was also detected at the tumor site, although the blood background was consistently high. The biodistribution results were consistent with the micro-SPECT/CT imaging results. 99mTc-MAG3-5-Fab could clearly display hTREM2+ A549 tumors in a short time (1 h) with low uptake in nontumor organs and tissues and thus has clinical application prospects.

Effect of rehabilitation therapy combined with neuromuscular electrical stimulation on cognitive dysfunction and activities of daily life in stroke patients.

OBJECTIVE: To determine the effects of rehabilitation therapy combined with neuromuscular electrical stimulation (NES) on cognitive dysfunction and ability to perform activities of daily living (ADLs) of stroke patients. METHODS: The clinical data of 100 stroke patients treated in the Second Affiliated Hospital of Soochow University from February 2019 to August 2021 were retrospectively analyzed. According to the therapeutic regimen, the patients given rehabilitation therapy combined with NES were assigned to a study group (n=52) and those given rehabilitation therapy alone were assigned to a control group (n=48). The treatment efficacy in the two groups was evaluated, and the levels of plasma cortisol (Cor) and neuropeptide Y (NPY), neurological function, motor function, balance ability, swallowing function, cognitive function, negative emotions, and quality of life (QoL) after therapy were evaluated. The maximum amplitude of surface electromyography (sEMG) and swallowing time were compared between the two groups. RESULTS: The study group yielded significantly better efficacy than the control group (P<0.05). Before therapy, there were no significant differences between the two groups in Cor and NPY levels, neurological function, motor function, balance ability, swallowing function, cognitive function, sEMG, swallowing time or negative emotions (P>0.05). After therapy, the above all indices all greatly improved, with more notso in the study group. In addition, after therapy, the study group had significantly better QoL indexes than the control group. CONCLUSION: Rehabilitation therapy combined with NES is effective in treating stroke. It can substantially ameliorate the cognitive dysfunction, prognosis and QoL in patients.

Nomogram Based on Dual-Layer Spectral Detector CTA Parameter for the Prediction of Infarct Core in Patients with Acute Ischemic Stroke.

(1) Background: Acute ischemic stroke (AIS) is time-sensitive. The accurate identification of the infarct core and penumbra areas in AIS patients is an important basis for formulating treatment plans, and is the key to dual-layer spectral detector computed tomography angiography (DLCTA), a safer and more accurate diagnostic method for AIS that will replace computed tomography perfusion (CTP) in the future. Thus, this study aimed to investigate the value of DLCTA in differentiating infarct core from penumbra in patients with AIS to establish a nomogram combined with spectral computed tomography (CT) parameters for predicting the infarct core and performing multi-angle evaluation. (2) Methods: Data for 102 patients with AIS were retrospectively collected. All patients underwent DLCTA and CTP. The patients were divided into the non-infarct core group and the infarct core group, using CTP as the reference. Multivariate logistic regression analysis was used to screen predictors related to the infarct core and establish a nomogram model. The receiver operating characteristic (ROC) curve, the calibration curve, and decision curve analysis (DCA) were used to evaluate the predictive efficacy, accuracy, and clinical practicability of the model, respectively. (3) Results: Multivariate logistic analysis identified three independent predictors: iodine density (OR: 0.022, 95% CI: 0.003-0.170, p < 0.001), hypertension (OR: 7.179, 95% CI: 1.766-29.186, p = 0.006), and triglycerides (OR: 0.255, 95% CI: 0.109-0.594, p = 0.002). The AUC-ROC of the nomogram was 0.913. Calibration was good. Decision curve analysis was clinically useful. (4) Conclusions: The spectral CT parameters, specifically iodine density values, effectively differentiate between the infarct core and penumbra areas in patients with AIS. The nomogram, based on iodine density values, showed strong predictive power, discrimination, and clinical utility to accurately predict infarct core in AIS patients.

The image quality and diagnostic performance of CT perfusion-derived CT angiography versus that of conventional CT angiography.

Background: The combination of computed tomography angiography (CTA) and computed tomography perfusion (CTP) evaluation of cerebral perfusion status and vascular conditions can improve the diagnostic accuracy of infarction, ischemia, and vascular occlusion in stroke patients, as well as a comprehensive assessment of cerebral edema, collateral circulation, and blood perfusion in the lesion area. However, the consequent radiation safety and contrast agent nephropathy have aroused increasing concern. The purpose of this study was to assess the image quality and diagnostic accuracy of CTA images derived from CTP data, and to explore the feasibility of replacing conventional CTA. Methods: A total of 31 consecutive patients with suspected acute ischemic stroke were retrospectively analyzed. All patients underwent head and neck CTA and brain CTP examinations. All the CTP images were transmitted to the ShuKun artificial intelligence system, which reconstructs CTA derived from CTP (CTA-DF-CTP). The images were divided into 2 groups, including CTA-DF-CTP (Group A) and conventional CTA (Group B). The CT attenuation values, subjective image noise, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), image quality, CT volume dose index (CTDIvol), dose length product (DLP), and effective radiation dose (ED) were compared between the 2 groups. Moreover, the consistency of vascular stenosis and stenosis degree between the 2 groups were measured and evaluated. Results: There were no significant differences in image noise, SNR, or CNR between Groups A and B (P>0.05). The CT attenuation values of the arteries were higher in Group A than in B [internal carotid artery (ICA) =548±112 vs. 454±85 Hounsfield units (HU), middle cerebral artery (MCA) =453±118 vs. 388±70 HU, and basilar artery (BA) =431±99 vs. 360±83 HU] (P<0.01). The image quality of the 2 groups met the requirement of clinical diagnosis (4.97±0.18 vs. 4.94±0.25). No significant difference was found in subjective evaluation (P>0.05). In Group A compared with Group B, the following reductions were observed: CTDIvol (10.7%; 100.8 vs. 112.9 mGy), DLP (23.0%; 1,613±0 vs. 2,093±88 mGy·cm), and ED (23.0%; 5.00±0.00 vs. 6.49±0.27 mSv). Conclusions: CTA-DF-CTP data provide diagnostic accuracy and image quality similar to those of conventional CTA of head and neck CTA.

Dynamic 68Ga-DOTA0-Tyr3-octreotate positron emission tomography-computed tomography for the evaluation of pancreatic neuroendocrine tumors: a pilot study.

Background: 68Ga-DOTA0-Tyr3-octreotate (68Ga-DOTATATE) is a radiolabeled somatostatin analog used for the diagnosis of pancreatic neuroendocrine tumors (pNETs), and standardized uptake value (SUV) measurements for therapeutic monitoring is recommended. However, changes in net influx rate (Ki) may better reflect treatment effects than may those of the SUV. The aim of this study was to investigate the value of dynamic 68Ga-DOTATATE positron emission tomography-computed tomography (PET-CT) in the evaluation of pNETs. Methods: Dynamic PET-CT scans over 60 min were acquired for 7 patients with localized pancreatic mass before surgery. Maximal and mean SUV (SUVmax and SUVmean) were measured in tumors and normal pancreatic body as reference tissue (RT). Time-activity curves (TACs) were extracted from tumors and RT. A 2-tissue compartment model was used to calculate the rate constants K1, k2, and k3 (min-1); Ki (mL/g/min); and K1:k2 ratio. The following statistical tests were used to evaluate the results: the Shapiro-Wilk, Student t test, Mann-Whitney, Spearman, and Pearson rank correlation tests. Results: Among 6 patients, 8 primary tumors were histopathologically proven to be pNETs. Moreover, 6 lesions with high uptake of 68Ga-DOTATATE showed an ascending TAC pattern, while 2 lesions with no or low uptake showed a descending TAC pattern. The mean SUVmax and SUVmean of pNETs were 46.4±40.2 (range, 3.9-109.9) and 21.9±16.0 (range, 0.5-42.8), respectively, which were significantly higher than the SUVmax of 4.2±0.6 (range, 3.1-4.9) and the SUVmean of 2.7±1.0 (range, 1.4-3.6) for the RT (P=0.021 and P=0.036), respectively. The Ki of pNETs was statistically higher than that of the RT [pNET: 0.366±0.372 (range, 0.019-0.992); RT: 0.060±0.017 (range, 0.04-0.08); P=0.036]. The mean K1:k2 ratio in pNETs was 12-fold higher than that of RT (6.06 vs. 0.50). In pNETs, there was a positive correlation between SUVmax and Ki (r=0.952; P<0.001) and between SUVmean and Ki (r=0.905; P=0.002). Another patient was diagnosed with intrapancreatic accessory spleen. Conclusions: The uptake of 68Ga-DOTATATE by pNETs can be explained by its high Ki value and K1:k2 ratio. Dynamic 68Ga-DOTATATE PET-CT can serve as a potential tool for evaluating pNETs and support the further assessment of a larger cohort of patients.

Low-frequency Electrical Stimulation of the Hippocampus Plays a Role in the Treatment of Pharmacoresistant Epilepsy by Blocking the PKA-CREB Pathway.

OBJECTIVE: The objective of this study is to study the mechanism of Low frequency electrical stimulation (LFS) in the treatment of drug-resistant epilepsy by regulating the protein kinase A (PKA)-cAMP response element-binding protein (CREB) signaling pathway upstream of gamma aminobutyric acid A (GABAA) receptor. METHODS: Primary hippocampal neurons were extracted and cultured from fetal rat brains and randomly divided into the normal control group, PKA-CREB agonist group, and PKA-CREB inhibitor group. Drug-resistant epileptic rats were established and randomly divided into the pharmacoresistant group, LFS group, PKA-CREB agonist combined with hippocampal LFS group, and PKA-CREB inhibitor combined with hippocampal LFS group. The normal rats were in the normal control group and drug-sensitive rats were in the pharmacosensitive group. The seizure frequency of epileptic rats was determined using video surveillance. The expression of PKA, CREB, p-CREB, and GABAA receptor subunits α1 and ß2 of each group were detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting assays. RESULTS: The in vitro expression levels of PKA, CREB, and p-CREB in the agonist group were significantly higher than those in the normal control group (NRC group), while the expression levels of GABAA receptor subunits α1 and ß2 were significantly lower than those in the NRC group. The expression levels of PKA, CREB, and p-CREB in the inhibitor group were significantly lower, while the expression levels of GABAA receptor subunits α1 and ß2 were significantly higher than those in the NRC group. The in vivo seizure frequency was significantly lower in the LFS group than in the pharmacoresistant group (PRE group). Compared to the LFS group, the seizure frequency and the expression levels of PKA, CREB, and p-CREB in the rat hippocampus were significantly higher, and the expression levels of GABAA receptor subunits α1 and ß2 were significantly lower in the agonist group. The results in the inhibitor group were exactly the opposite of those in the agonist group. CONCLUSION: The PKA-CREB signaling pathway is involved in the regulation of GABAA receptor subunits α1 and ß2. In addition, LFS plays an important role in increasing GABAA receptor expression by regulating the PKA-CREB signaling pathway.

Association of APOE-ε4, Osteoarthritis, ß-Amyloid, and Tau Accumulation in Primary Motor and Somatosensory Regions in Alzheimer Disease.

BACKGROUND AND OBJECTIVES: One of the most prevalent chronic diseases, osteoarthritis (OA), may work in conjunction with APOE-ε4 to accelerate Alzheimer disease (AD) alterations, particularly in the primary motor (precentral) and somatosensory (postcentral) cortices. To understand the reasoning behind this, we investigated how OA and APOE-ε4 influence the accumulation of ß-amyloid (Aß) and tau accumulation in primary motor and somatosensory regions in Aß-positive (Aß+) older individuals. METHODS: We selected Aß+ Alzheimer Disease Neuroimaging Initiative participants, defined by baseline 18F-florbetapir (FBP) Aß PET standardized uptake value ratio (SUVR) of AD summary cortical regions, who had longitudinal Aß PET, the records of OA medical history, and APOE-ε4 genotyping. We examined how OA and APOE-ε4 relate to baseline and longitudinal Aß accumulation and tau deposition measured at follow-up in precentral and postcentral cortical areas and how they modulate Aß-associated future higher tau levels, adjusting for age, sex, and diagnosis and using multiple comparison corrections. RESULTS: A total of 374 individuals (mean age 75 years, 49.2% female, 62.8% APOE-ε4 carriers) who underwent longitudinal FBP PET with a median follow-up of 3.3 years (interquartile range [IQR] 3.4, range 1.6-9.4) were analyzed, and 96 people had 18F-flortaucipir (FTP) tau PET measured at a median of 5.4 (IQR 1.9, range 4.0-9.3) years postbaseline FBP PET. Neither OA nor APOE-ε4 was related to baseline FBP SUVR in precentral and postcentral regions. At follow-up, OA rather than APOE-ε4 was associated with faster Aß accumulation in postcentral region (ß = 0.005, 95% CI 0.001-0.008) over time. In addition, OA but not the APOE-ε4 allele was strongly linked to higher follow-up FTP tau levels in precentral (ß = 0.098, 95% CI 0.034-0.162) and postcentral (ß = 0.105, 95% CI 0.040-0.169) cortices. OA and APOE-ε4 were also interactively associated with higher follow-up FTP tau deposition in precentral (ß = 0.128, 95% CI 0.030-0.226) and postcentral (ß = 0.124, 95% CI 0.027-0.223) regions. DISCUSSION: This study suggests that OA was associated with faster Aß accumulation and higher Aß-dependent future tau deposition in primary motor and somatosensory regions, providing novel insights into how OA increases the risk of AD.

Radiomics model using preoperative computed tomography angiography images to differentiate new from old emboli of acute lower limb arterial embolism.

Our purpose was to devise a radiomics model using preoperative computed tomography angiography (CTA) images to differentiate new from old emboli of acute lower limb arterial embolism. 57 patients (95 regions of interest; training set: n = 57; internal validation set: n = 38) with femoral popliteal acute lower limb arterial embolism confirmed by pathology and with preoperative CTA images were retrospectively analyzed. We selected the best prediction model according to the model performance tested by area under the curve (AUC) analysis across 1,000 iterations of prediction from three most common machine learning methods: support vector machine, feed-forward neural network (FNN), and random forest, through several steps of feature selection. Then, the selected best model was also validated in an external validation dataset (n = 24). The established radiomics signature had good predictive efficacy. FNN exhibited the best model performance on the training and validation groups: its AUC value was 0.960 (95% CI, 0.899-1). The accuracy of this model was 89.5%, and its sensitivity and specificity were 0.938 and 0.864, respectively. The AUC of external validation dataset was 0.793. Our radiomics model based on preoperative CTA images is valuable. The radiomics approach of preoperative CTA to differentiate new emboli from old is feasible.

Diagnostic value of MRI perfusion-weighted imaging and diffusion-weighted imaging parameters in cerebral apoplexy.

OBJECTIVE: To assess the clinical and prognostic value of MRI perfusion-weighted imaging (PWI) versus diffusion-weighted imaging (DWI) parameters for apparent diffusion coefficient (ADC) and cerebral blood flow (CBF) in the diagnosis of patients with ischemic stroke (IS). METHODS: Eighty patients diagnosed with IS in the Second Affiliated Hospital of Soochow University from January 2020 to January 2021 were retrospectively analyzed and regarded as a patient group. Meanwhile, 50 patients who underwent physical examination at the Second Affiliated Hospital of Soochow University during the same period were collected, and were identified by physical examination to have atherosclerotic stenosis but not cerebral infarction, they were set as a control group. The differences of ADC and CBF between both groups were compared. The diagnostic value of ADC and CBF in diagnosing acute ischemic stroke was analyzed by receiver operating characteristic (ROC) curve. The changes of ADC and CBF before and after treatment were compared. Patients were sub-grouped according to their mRS scores, and those with scores of 0-2 were grouped into the good prognosis group while those with scores of 3-6 were grouped into the poor prognosis group, and the risk factors affecting patients' prognosis were evaluated by logistic regression. The correlation of ADC and CBF with National Institutes of Health Stroke Scale (NIHSS) scores and modified Rankin Scale (mRS) scores was analyzed. ADC and CBF levels were compared between deceased and surviving patients, and their predictive value was assessed by ROC curves. RESULTS: ADC and CBF were dramatically lower in the patient group compared with the control group (P<0.05). The area under the curve (AUC) of ADC and CBF in diagnosing IS was 0.949 and 0.926. The ADC and CBF values after treatment were significantly increased as compared to before treatment (both P<0.05). Both ADC and CBF were lower in the patients of the deceased group than in those in the survival group (P<0.05). The AUC for ADC and CBF in predicting death in patients diagnosed with IS was 0.866 and 0.766, respectively. ADC, CBF was negatively correlated with patients' NIHSS and mRS scores (P<0.01). Higher pre-treatment NIHSS and admission time ≥24 h after onset were risk factors for patient prognosis, whereas higher ADC and CBF values were protective factors (all P<0.05). CONCLUSION: ADC and CBF values are reduced in IS patients and can be used as a diagnostic and prognostic indicator of IS.

124I-Labeled Immuno-PET Targeting hTREM2 for the Diagnosis of Gastric Carcinoma.

Low ß-2-[18F]-fluoro-2-deoxy-d-glucose (18F-FDG) uptake in gastric mucinous adenocarcinoma may cause false-negative diagnosis and erroneous staging. Thus, there is an urgent need for developing tumor-specific imaging agents in gastric cancer diagnostics. Triggering receptor expressed on myeloid cells 2 (TREM2) is a transmembrane protein expressed on the surface of tumor-associated macrophages (TAMs) and is considerably overexpressed in tumor tissues. This study aimed to develop new human TREM2 (hTREM2)-targeting imaging agents to diagnose and monitor gastric cancer. We established a cell line, MGC803, with upregulated expression of hTREM2, at the cell surface. We produced a monoclonal antibody (5-mAb) against hTREM2 by immunizing mice with the hTREM2 antigen to obtain the antibody fragment 5-F(ab')2 using an immunoglobulin G-degrading enzyme of Streptococcus pyogenes (IdeS). Another anti-TREM2-mAb (clone 237920) and its fragment anti-TREM2-F(ab')2 were employed for the comparative study in vitro and in vivo. After 124I labeling, we constructed the probes: 124I-5-mAb, 124I-5-F(ab')2, 124I-anti-TREM2-mAb, and 124I-anti-TREM2-F(ab')2. We found that 5-mAb exhibited higher hTREM2 affinity and slower blood clearance than anti-TREM2-mAb, whose corresponding F(ab')2 fragments demonstrated the same trend. The micro-PET/CT revealed that 124I-5-F(ab')2 exhibited advantages of tumor enrichment and fast metabolism. The biodistribution study results were consistent with those of micro-PET/CT. Among the four tracers, 124I-5-F(ab')2 was the most suitable specific radiotracer for targeting hTREM2 and displayed potential utility as a tumor-imaging tracer for diagnosing gastric carcinoma.

Exploration of 68Ga-DOTA-MAL as a Versatile Vehicle for Facile Labeling of a Variety of Thiol-Containing Bioactive Molecules.

Efficient and site-specific radiolabeling reactions are essential in molecular probe synthesis. Thus, selecting an effective method for radiolabeling that does not affect bioactivity of the molecule is critical. Varieties of bifunctional chelating agents provide a solution in this matter. As a chemo-specific chelator, maleimido-mono-amide-DOTA (DOTA-Mal) holds significant potential for 68Ga labeling of bioactive molecules; it can react specifically with free sulfhydryl groups under mild conditions. Compared with amino and carboxylic acid groups, free sulfhydryl groups are relatively less common in most biomolecules and can serve as site-specific radiolabeling targets. Labeling of 68Ga usually employs a two-step labeling strategy; first, chelators are conjugated to the biomolecules, which is followed by radiolabeling. However, the bioactivity of biomolecules may be affected by harsh labeling conditions. In this study, three 68Ga-labeled bioactive molecules, namely, 68Ga-DOTA-RGD, 68Ga-DOTA-FA, and 68Ga-DOTA-BSA, were prepared using a novel strategy under mild conditions (pH of 8.0 at room temperature). Using this strategy, DOTA-Mal was labeled by 68Ga before it reacted with the sulfhydryl group-containing biomolecules, which avoided damage to said biomolecules caused by the harsh reaction conditions required in 68Ga-labeling procedures. The biological and chemical properties of these three radiotracers synthesized using this strategy are well manifested. Through a series of experiments, the effectiveness of this strategy is demonstrated, and we believe that this site-specific bioactivity-friendly reaction strategy will facilitate developments and translation applications of varieties of 68Ga-labeled positron emission tomography probes.

Noninvasive evaluation of PD-L1 expression in non-small cell lung cancer by immunoPET imaging using an acylating agent-modified antibody fragment.

PURPOSE: The aim of this study was to explore an effective 124I labeling strategy and improve the signal-to-noise ratio when evaluating the expression of PD-L1 using an 124I-iodinated durvalumab (durva) F(ab')2 fragment. METHODS: The prepared durva F(ab')2 fragments were incubated with N-succinimidyl-3-(4-hydroxyphenyl) propionate (SHPP); after purification, the HPP-durva F(ab')2 was iodinated using Iodo-Gen method. After the radiochemical purity, stability, and specific activities were determined, the binding affinities of probes prepared using different labeling strategies were compared in vitro. The clinical application value of [124I]I-HPP-durva-F(ab')2 was confirmed by PET imaging. To more objectively evaluate the in vivo distribution and clearance of tracers, the pharmacokinetics and biodistribution assays were also performed. RESULTS: After being modified with SHPP, the average conjugation number of SHPP per durva-F(ab')2 identified by LC-MS was about 8.92 ± 2.84. The prepared [124I]I-HPP-durva F(ab')2 was obtained with a satisfactory radiochemical purity of more than 98% and stability of more than 93% when incubated for 72 h. Compared with unmodified [124I]I-durva F(ab')2, the specific activity of [124I]I-HPP-durva-F(ab')2 was improved (52.91 ± 5.55 MBq/mg and 15.91 ± 0.74 MBq/mg), while the affinity did not significantly change. The biodistribution experiments and PET imaging showed that the prepared [124I]I-HPP-durva-F(ab')2 exhibited an accelerated clearance and improved tumor-to-background ratio compared with [124I]I-durva-F(ab')2. The specificity of [124I]I-HPP-durva-F(ab')2 to PD-L1 was well demonstrated both in vitro and in vivo. CONCLUSIONS: A PD-L1 PET imaging probe [124I]I-HPP-durva F(ab')2 was successfully synthesized through the SHPP modification strategy. The prepared probe was able to accurately evaluate the PD-L1 expression level through high-contrast noninvasive imaging.

The Anaesthetics Isoflurane and Xenon Reverse the Synaptotoxic Effects of Aß1-42 on Megf10-Dependent Astrocytic Synapse Elimination and Spine Density in Ex Vivo Hippocampal Brain Slices.

It has been hypothesised that inhalational anaesthetics such as isoflurane (Iso) may trigger the pathogenesis of Alzheimer's disease (AD), while the gaseous anaesthetic xenon (Xe) exhibits many features of a putative neuroprotective agent. Loss of synapses is regarded as one key cause of dementia in AD. Multiple EGF-like domains 10 (MEGF10) is one of the phagocytic receptors which assists the elimination of synapses by astrocytes. Here, we investigated how ß-amyloid peptide 1-42 (Aß1-42), Iso and Xe interact with MEGF10-dependent synapse elimination. Murine cultured astrocytes as well as cortical and hippocampal ex vivo brain slices were treated with either Aß1-42, Iso or Xe and the combination of Aß1-42 with either Iso or Xe. We quantified MEGF10 expression in astrocytes and dendritic spine density (DSD) in slices. In brain slices of wild type and AAV-induced MEGF10 knock-down mice, antibodies against astrocytes (GFAP), pre- (synaptophysin) and postsynaptic (PSD95) components were used for co-localization analyses by means of immunofluorescence-imaging and 3D rendering techniques. Aß1-42 elevated pre- and postsynaptic components inside astrocytes and decreased DSD. The combined application with either Iso or Xe reversed these effects. In the presence of Aß1-42 both anaesthetics decreased MEGF10 expression. AAV-induced knock-down of MEGF10 reduced the pre- and postsynaptic marker inside astrocytes. The presented data suggest Iso and Xe are able to reverse the Aß1-42-induced enhancement of synaptic elimination in ex vivo hippocampal brain slices, presumably through MEGF10 downregulation.