Simultaneous multimodal three-photon and optical coherence microscopy of the mouse brain in the 1700 nm optical window in vivo.

Multimodal microscopy combining various imaging approaches can provide complementary information about tissue in a single imaging session. Here, we demonstrate a multimodal approach combining three-photon microscopy (3PM) and spectral-domain optical coherence microscopy (SD-OCM). We show that an optical parametric chirped-pulse amplification (OPCPA) laser source, which is the standard source for three-photon fluorescence excitation and third harmonic generation (THG), can be used for simultaneous OCM, 3-photon (3P) fluorescence and THG imaging. We validated the system performance in deep mouse brains in vivo with an OPCPA source operating at 1620 nm center wavelength. We visualized small structures such as myelinated axons, neurons, and large fiber tracts in white matter with high spatial resolution non-invasively using linear and nonlinear contrast at >1 mm depth in intact adult mouse brain. Our results showed that simultaneous OCM and 3PM at the long wavelength window can be conveniently combined for deep tissue imaging in vivo.

Comparison of Baseline 68Ga-FAPI and 18F-FDG PET/CT for Prediction of Response and Clinical Outcome in Patients with Unresectable Hepatocellular Carcinoma Treated with PD-1 Inhibitor and Lenvatinib.

Fibroblast activation protein contributes to immunosuppression and resistance to immunotherapies. This study aimed to compare baseline 68Ga-labeled fibroblast activation protein inhibitor (68Ga-FAPI) PET/CT and 18F-FDG PET/CT in response and survival prediction in unresectable hepatocellular carcinoma (uHCC) patients treated with the combination of programmed cell death 1 (PD-1) inhibitor and lenvatinib. Methods: In this prospective cohort study, 22 patients with uHCC who underwent baseline 18F-FDG and 68Ga-FAPI PET/CT and soon began taking a combination of PD-1 inhibitor and lenvatinib were recruited. Semiquantitative indices of baseline PET/CT were measured as 18F-FDG SUVmax, metabolic tumor volume, total lesion glycolysis, 68Ga-FAPI SUVmax, 68Ga-FAPI-avid tumor volume (FTV), and total lesion fibroblast activation protein expression (TLF). The primary endpoint was durable or nondurable clinical benefit after treatment, and the secondary endpoints were progression-free survival (PFS) and overall survival (OS). Results: The overall response rate of the combination therapy was 41% (9/22). Fifty percent of patients had durable clinical benefit. Median PFS and OS were 4.8 and 14.4 mo, respectively. Patients with nondurable clinical benefit showed a significantly higher FTV and TLF than those with durable clinical benefit, whereas 18F-FDG parameters overlapped. A higher 68Ga-FAPI-avid tumor burden (FTV > 230.46 cm3 or TLF > 961.74 SUVbody weight⋅cm3) predicted both shorter PFS (4.0 vs. 13.5 mo, P = 0.016) and shorter OS (7.8 mo vs. not reached, P = 0.030). Patients with a higher metabolic tumor burden (metabolic tumor volume > 206.80 cm3 or total lesion glycolysis > 693.53 SUVbody weight⋅cm3) showed a shorter OS although the difference did not reach statistical significance (P = 0.085). In multivariate analysis, a higher 68Ga-FAPI-avid tumor burden (hazard ratio [HR], 3.88 [95% CI, 1.26-12.01]; P = 0.020) and macrovascular invasion (HR, 4.00 [95% CI, 1.06-15.14]; P = 0.039) independently predicted a shorter PFS, whereas a higher 68Ga-FAPI-avid tumor burden (HR, 5.92 [95% CI, 1.19-29.42]; P = 0.035) and bone metastases (HR, 5.88 [95% CI, 1.33-25.93]; P = 0.022) independently predicted a shorter OS. Conclusion: Volumetric indices on baseline 68Ga-FAPI PET/CT were potentially independent prognostic factors to predict durable clinical benefit, PFS, and OS in uHCC patients treated with a combination of PD-1 and lenvatinib. Baseline 68Ga-FAPI PET/CT may facilitate uHCC patient selection before combination therapy.

Metabolic Tumor Volume Measured by 18F-FDG PET/CT is Associated with the Survival of Unresectable Hepatocellular Carcinoma Treated with PD-1/PD-L1 Inhibitors Plus Molecular Targeted Agents.

Purpose: The combination of PD-1/PD-L1 inhibitors and molecular targeted agents showed promising efficacy for unresectable hepatocellular carcinoma (uHCC). This study aimed to investigate the prognostic value of metabolic parameters from 18F-fluorodeoxyglucose positron emission tomography-computed tomography (18F-FDG PET/CT) in patients with uHCC underwent the combined therapies. Patients and Methods: Patients with uHCC treated with a combination of immunotherapy and targeted therapy who underwent baseline 18F-FDG PET/CT between July 2018 and December 2021 were recruited retrospectively. The metabolic tumor volume (MTV), total lesion glycolysis (TLG), maximum standardized uptake values (SUVmax), and clinical and biological parameters were recorded. A multivariate prediction model was developed for overall survival (OS) using these parameters together with clinical prognostic factors. Results: Seventy-seven patients were finally included. The median OS was 16.8 months. We found that a high MTV (≥39.65 cm3 as the median value) was significantly associated with OS (P<0.05). In multivariate analyses for OS, a high MTV, high Eastern Cooperative Oncology Group performance status (ECOG-PS, ≥1), Child-Pugh (B-C) grade, and the presence of bone metastasis were significantly associated with poor OS (HR 1.371, HR 3.73, HR 15.384, and HR 2.994, all P<0.05, respectively). A multivariate prognostic model including MTV and prognostic factors, such as ECOG-PS, Child-Pugh grade, and bone metastasis, further improved the identification of different OS subgroups. Conclusion: High MTV is an adverse prognostic factor in patients with uHCC treated with a combination of immunotherapy and molecular targeted agents. Integrating PET/CT parameters with clinical prognostic factors could help to personalize immunotherapy.

Generalized Muscular Hypermetabolism Caused by Mitochondrial Myopathy Shown on 18 F-FDG PET/CT.

ABSTRACT: An 18-year-old man presented with progressive exercise intolerance and muscle weakness for 1 year with recent acute exacerbation. Laboratory test demonstrated lactic acidosis. 18 F-FDG PET/CT was performed to exclude malignancy and showed generalized muscular hypermetabolism. Muscle biopsy combined with patient's history suggested mitochondrial myopathy. This report illustrates that mitochondrial myopathy may present as generalized muscular hypermetabolism on 18 F-FDG PET/CT and thus should be added to the differential diagnoses.

Different Uptake of 68Ga-FAPI and 18F-FDG in Lymphadenopathy Caused by Angioimmunoblastic T-Cell Lymphoma in a Patient with Colon Cancer.

An 82-year-old man with a history of colon cancer was found with multiple lymphadenopathies and a pulmonary mass. Fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) detected generalized hypermetabolic lymph nodes and an FDG-avid pulmonary mass. PET/CT with 68Ga-labeled fibroblast activation protein inhibitor (FAPI) revealed intense uptake in the lung mass, consistent with metastatic disease from colon cancer. However, the lymphadenopathies were not avid for 68Ga-FAPI, suggesting a different etiology. The biopsy of a cervical node confirmed angioimmunoblastic T-cell lymphoma. The case showcased the potential of 68Ga-FAPI in differentiation of solid tumor and hematological disease due to different histopathologic nature of stromal fibrosis.

Enhanced brassinosteroid signaling via the overexpression of SlBRI1 positively regulates the chilling stress tolerance of tomato.

Brassinosteroids (BRs) regulate plant development and response to stress. BRASSINOSTEROID INSENSITIVE 1 (BRI1) is a BR receptor that activates BR signaling. Although the function of the tomato BR receptor SlBRI1 in regulating growth and drought resistance has been examined, that of SlBRI1 in cold tolerance is unclear. This study indicated that the expression of SlBRI1 in tomato was rapidly induced and reached its highest level at 3 h under chilling treatment and then decreased. The overexpression of SlBRI1 displayed low relative electrolyte leakage, malondialdehyde content, and reactive oxygen species (ROS) accumulation under chilling stress. The proline content and activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in SlBRI1OE plants were higher than those in the wild-type (WT) plants after chilling stress. The transcript abundances of five cold-responsive genes were higher in SlBRI1OE plants than in WT plants during chilling stress. RNA sequence analysis showed that the expression of the majority of genes encoding photosystem I and II were downregulated. The degree of suppression in SlBRI1OE plants was weaker than that in WT plants. Additionally, the Pn and Fv/Fm of SlBRI1OE plants were significantly higher than those of WT plants under chilling stress. We identified several genes encoding key enzymes in BRs; indole acetic acid (IAA), gibberellin (GA), and abscisic acid (ABA) biosynthesis or signaling were upregulated or downregulated during chilling stress. Chilling stress decreased the BRs and GA3 content, and increased IAA and ABA content. The contents were lower or higher in SlBRI1OE than in WT plants. Furthermore, chilling stress regulated the expression levels of 43 transcription factors. The expression of seven cold-regulated protein genes was higher or lower in SlBRI1OE plants than in WT plants under chilling stress. These results suggest that SlBRI1 positively regulates chilling tolerance mainly through ICE1-CBF-COR pathway in tomato.

Complementary Utility of Dopamine Transporter and Tau PET Imaging in the Diagnosis of Progressive Supranuclear Palsy: A Case Report.

ABSTRACT: A 50-year-old woman developed gait disturbances, tendency to fall backwards, bradykinesia, and memory loss over the previous 6 months. Brain 18F-FDG PET/CT was unable to distinguish among APSs (atypical parkinsonian syndromes); PET investigations of dopamine transporter (DAT) function (11C-CFT) and tau pathology (18F-APN-1607) were performed. 11C-CFT PET revealed severe loss of striatal DAT function, whereas significant tau accumulation was observed in the brainstem, basal ganglia, and globus pallidus on 18F-APN-1607 PET. Such finding suggested diagnosis of PSP (progressive supranuclear palsy). This case highlights the value of DAT and tau PET imaging in diagnosis of PSP and differential diagnosis ofAPSs.

Feasibility of In Vivo Imaging of Fibroblast Activation Protein in Human Arterial Walls.

Increased expression of fibroblast-activating protein (FAP) in fibrous caps may contribute to progression of atherosclerotic plaques. Methods: Forty-one patients who underwent 68Ga-conjugated quinoline-based FAP inhibitor (68Ga-FAPI-04) PET/CT for noncardiovascular indications were retrospectively analyzed. Correlations were assessed between the uptake of 68Ga-FAPI-04 in large arterial walls (SUVmax and target-to-background ratio, TBR) and degree of calcification and cardiovascular risk factors. Results: Focal arterial uptake of 68Ga-FAPI-04 or calcification was detected in 1,177 arterial segments in all 41 patients. TBR was negatively correlated with the degree of calcification (Hounsfield units) (r = -0.27, P < 0.01). Mean TBR in higher-risk patients was greater than in lower-risk patients (2.2 ± 0.3 vs. 1.8 ± 0.3, P < 0.01). Immunohistochemical labeling of carotid plaques exhibited prominent FAP expression in a thin fibrous cap and moderate FAP expression in a thick cap. Conclusion:68Ga-FAPI-04 PET/CT might have potential for imaging fibroblastic activation in the arterial wall.

Closed-loop wavefront sensing and correction in the mouse brain with computed optical coherence microscopy.

Optical coherence microscopy (OCM) uses interferometric detection to capture the complex optical field with high sensitivity, which enables computational wavefront retrieval using back-scattered light from the sample. Compared to a conventional wavefront sensor, aberration sensing with OCM via computational adaptive optics (CAO) leverages coherence and confocal gating to obtain signals from the focus with less cross-talk from other depths or transverse locations within the field-of-view. Here, we present an investigation of the performance of CAO-based aberration sensing in simulation, bead phantoms, and ex vivo mouse brain tissue. We demonstrate that, due to the influence of the double-pass confocal OCM imaging geometry on the shape of computed pupil functions, computational sensing of high-order aberrations can suffer from signal attenuation in certain spatial-frequency bands and shape similarity with lower order counterparts. However, by sensing and correcting only low-order aberrations (astigmatism, coma, and trefoil), we still successfully corrected tissue-induced aberrations, leading to 3× increase in OCM signal intensity at a depth of ∼0.9 mm in a freshly dissected ex vivo mouse brain.

Ectopic Parathyroid Adenoma on Pectoralis Major After Endoscopic Parathyroidectomy Revealed by 99mTc-MIBI SPECT/CT.

ABSTRACT: A 45-year-old woman, who had endoscopic parathyroidectomy 5 years ago for a right inferior parathyroid adenoma, underwent 99mTc-MIBI scan for the evaluation of recurrent parathyroidism. The images did not identify any abnormal uptake in the neck. Instead, an abnormal activity in the right upper chest wall was noted, which was located in a soft tissue nodule inseparable from the right pectoralis major muscle on SPECT/CT images. Pathology confirmed parathyroid tissue.

Hypertrophic spinal pachymeningitis caused by ANCA-associated vasculitis revealed by 18F-FDG PET/CT: A case report.

RATIONALE: Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) can involve the central nervous system in estimatedly 15% of patients. Hypertrophic pachymeningitis causes inflammatory hypertrophy of the cranial or spinal dura mater and patients present with various neurological deficits. ANCA-associated hypertrophic spinal pachymeningitis has rarely been reported in literature. We report a case of AAV presenting with hypertrophic spinal pachymeningitis detected by 18F-FDG PET/CT. PATIENT CONCERNS: A 66-year-old woman diagnosed with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis developed back pain, bilateral lower limb weakness, dysuria, and dysporia 1 month ago. DIAGNOSIS: Contrast-enhanced MRI showed thickening and enhancement of the dura mater in the thoracic cord. Intraspinal hypermetabolism in the corresponding region was observed on 18F-FDG PET/CT. The patient was finally diagnosed with ANCA-associated hypertrophic spinal pachymeningitis. INTERVENTIONS: The patient was treated with a higher dose of prednisone and cyclophosphamide. OUTCOMES: After 2-week treatment, the patient's neurological symptoms improved rapidly and laboratory findings were ameliorated. A repeated contrast-enhanced MRI showed partial improvement of the disease in the thoracic cord. LESSONS: 18F-FDG PET/CT and contrast-enhanced MRI can aid in the clinical diagnosis and surveillance in AAV-associated hypertrophic spinal pachymeningitis and potentially facilitate early recognition and intervention to prevent irreversible neurological impairment.

Performance of PET imaging for the localization of epileptogenic zone in patients with epilepsy: a meta-analysis.

OBJECTIVES: The aim of this meta-analysis was to estimate the clinical use value of 11C-FMZ and 18F-FDG in PET for the localization of epileptogenic zone and to provide evidence for practitioners' clinical decision-making. METHODS: We searched PubMed and Embase in a time frame from inception to May 31, 2020. Studies utilizing FMZ or FDG-PET or FDG-PET/MRI used in patients with epilepsy, with EEG or surgical outcomes as the gold standard and corresponding outcomes such as concordance rates of PET or PET/MRI scan compared with reference standard, absolute numbers of participants with true-positive (TP), false-positive (FP), true-negative (TN), and false-negative (FN) results in FDG or FMZ PET. Pooled concordance rates, overall sensitivity, and specificity of 11C-FMZ-PET and 18F-FDG-PET were calculated. RESULTS: In total, 44 studies met the inclusion criteria. The pooled concordance rates of FDG-PET, FMZ-PET, and FDG-PET/MRI coregistration compared with reference standard were 0.67 (95% CI: 0.60-0.73), 0.75 (95% CI: 0.57-0.93), and 0.93 (95% CI: 0.89-0.97), respectively. The concordance rate of 18F-FDG-PET in patients with temporal lobe epilepsy (TLE) was 0.79 (0.63; 0.92). The overall sensitivity and specificity of 18F-FDG-PET were 0.66 (95% CI: 0.58-0.73) and 0.71 (95% CI: 0.63-0.78), respectively. 11C-FMZ-PET displayed an overall sensitivity of 0.62 (95% CI: 0.49-0.73) and specificity of 0.73 (95% CI: 0.59-0.84). CONCLUSIONS: Both 11C-FMZ PET and 18F-FDG PET are the choice of modalities for the localization of epileptogenic zone, especially when coregistered with MRI. KEY POINTS: • 11C-FMZ-PET may be more helpful than 18F-FDG-PET in the localization of epilepsy foci. • Coregistration of FDG-PET and MRI is recommended in the localization of epileptogenic zone.

Investigation of multiple scattering in space and spatial-frequency domains: with application to the analysis of aberration-diverse optical coherence tomography.

Optical microscopy suffers from multiple scattering (MS), which limits the optical imaging depth into scattering media. We previously demonstrated aberration-diverse optical coherence tomography (AD-OCT) for MS suppression, based on the principle that for datasets acquired with different aberration states of the imaging beam, MS backgrounds become decorrelated while single scattering (SS) signals remain correlated, so that a simple coherent average can be used to enhance the SS signal over the MS background. Here, we propose a space/spatial-frequency domain analysis framework for the investigation of MS in OCT, and apply the framework to compare AD-OCT (using astigmatic beams) to standard Gaussian-beam OCT via experiments in scattering tissue phantoms. Utilizing this framework, we found that increasing the astigmatic magnitude produced a large drop in both MS background and SS signal, but the decay experienced by the MS background was larger than the SS signal. Accounting for the decay in both SS signal and MS background, the overall signal-to-background ratio (SBR) of AD-OCT was similar to the Gaussian control after about 10 coherent averages, when deeper line foci was positioned at the plane-of-interest and the line foci spacing was smaller than or equal to 80 µm. For an even larger line foci spacing of 160 µm, AD-OCT resulted in a lower SBR than the Gaussian-beam control. This work provides an analysis framework to gain deeper levels of understanding and insights for the future study of MS and MS suppression in both the space and spatial-frequency domains.

Secoisolariciresinol Diglucoside Improves Ovarian Reserve in Aging Mouse by Inhibiting Oxidative Stress.

Ovarian reserve is a key factor in the reproductive function of the ovaries. Ovarian aging is characterized by a gradual decline in the quantity and quality of follicles. The underlying mechanism of ovarian aging is complex and age-related oxidative stress is considered one of the most likely factors. Secoisolariciresinol diglucoside (SDG) has been shown to have good scavenging ability against reactive oxygen species (ROS) which slowly accumulates in ovarian tissues. However, it is unknown whether SDG had beneficial effects on aging ovaries. In this study, we used 37-week-old female C57BL/6J mouse as a natural reproductive aging model to evaluate the role of SDG in ovarian aging. SDG (7 and 70 mg/kg) intragastric administration was performed in the mice daily. After 8 weeks, the effects of SDG on aging ovaries were evaluated by counting the number of follicles and the expression of follicle-stimulating hormone receptors (FSHR) in the ovary. The mechanism of SDG on the aging ovaries was further explored through ovarian metabolomics. It was found that SDG can effectively increase the number of growing follicles and increase the expression of the FSHR protein. The metabolomics results showed that the ovaries in the SDG intervention group achieved better uptake and transport of nutrients, including amino acids and glucose that are necessary for the development of oocytes. At the same time, the ovaries of the SDG intervention group showed that the drug reduced ROS generation. Additionally, we found that ovarian telomere length and ovarian mitochondrial DNA copy number that are highly susceptible to ROS damage and are also related to aging. The results showed that SDG can significantly increase mitochondrial DNA copy number and slow down the process of telomere shortening. These data indicate that SDG improves ovarian reserve by inhibiting oxidative stress.

Extensive Intraspinal Hypermetabolism Caused by Neurobrucellosis Shown on 18F-FDG PET/CT.

A 55-year-old man and a 52-year-old man presenting with neurological symptoms were referred for F-FDG PET/CT for the underlying malignancy. In both cases, extensive intraspinal hypermetabolism was observed in F-FDG PET/CT. The cerebrospinal fluid culture finally confirmed infection of Brucella. This report illustrates that neurobrucellosis may present as intraspinal hypermetabolism on F-FDG PET/CT, and thus it should be added to the differential diagnosis.

Computed optical coherence microscopy of mouse brain ex vivo.

The compromise between lateral resolution and usable imaging depth range is a bottleneck for optical coherence tomography (OCT). Existing solutions for optical coherence microscopy (OCM) suffer from either large data size and long acquisition time or a nonideal point spread function. We present volumetric OCM of mouse brain ex vivo with a large depth coverage by leveraging computational adaptive optics (CAO) to significantly reduce the number of OCM volumes that need to be acquired with a Gaussian beam focused at different depths. We demonstrate volumetric reconstruction of ex-vivo mouse brain with lateral resolution of 2.2 µm, axial resolution of 4.7 µm, and depth range of ∼1.2 mm optical path length, using only 11 OCT data volumes acquired on a spectral-domain OCM system. Compared to focus scanning with step size equal to the Rayleigh length of the beam, this is a factor of 4 fewer datasets required for volumetric imaging. Coregistered two-photon microscopy confirmed that CAO-OCM reconstructions can visualize various tissue microstructures in the brain. Our results also highlight the limitations of CAO in highly scattering media, particularly when attempting to reconstruct far from the focal plane or when imaging deep within the sample.

A deep learning method to more accurately recall known lysine acetylation sites.

BACKGROUND: Lysine acetylation in protein is one of the most important post-translational modifications (PTMs). It plays an important role in essential biological processes and is related to various diseases. To obtain a comprehensive understanding of regulatory mechanism of lysine acetylation, the key is to identify lysine acetylation sites. Previously, several shallow machine learning algorithms had been applied to predict lysine modification sites in proteins. However, shallow machine learning has some disadvantages. For instance, it is not as effective as deep learning for processing big data. RESULTS: In this work, a novel predictor named DeepAcet was developed to predict acetylation sites. Six encoding schemes were adopted, including a one-hot, BLOSUM62 matrix, a composition of K-space amino acid pairs, information gain, physicochemical properties, and a position specific scoring matrix to represent the modified residues. A multilayer perceptron (MLP) was utilized to construct a model to predict lysine acetylation sites in proteins with many different features. We also integrated all features and implemented the feature selection method to select a feature set that contained 2199 features. As a result, the best prediction achieved 84.95% accuracy, 83.45% specificity, 86.44% sensitivity, 0.8540 AUC, and 0.6993 MCC in a 10-fold cross-validation. For an independent test set, the prediction achieved 84.87% accuracy, 83.46% specificity, 86.28% sensitivity, 0.8407 AUC, and 0.6977 MCC. CONCLUSION: The predictive performance of our DeepAcet is better than that of other existing methods. DeepAcet can be freely downloaded from https://github.com/Sunmile/DeepAcet .

LipoSVM: Prediction of Lysine Lipoylation in Proteins based on the Support Vector Machine.

BACKGROUND: Lysine lipoylation which is a rare and highly conserved post-translational modification of proteins has been considered as one of the most important processes in the biological field. To obtain a comprehensive understanding of regulatory mechanism of lysine lipoylation, the key is to identify lysine lipoylated sites. The experimental methods are expensive and laborious. Due to the high cost and complexity of experimental methods, it is urgent to develop computational ways to predict lipoylation sites. METHODOLOGY: In this work, a predictor named LipoSVM is developed to accurately predict lipoylation sites. To overcome the problem of an unbalanced sample, synthetic minority over-sampling technique (SMOTE) is utilized to balance negative and positive samples. Furthermore, different ratios of positive and negative samples are chosen as training sets. RESULTS: By comparing five different encoding schemes and five classification algorithms, LipoSVM is constructed finally by using a training set with positive and negative sample ratio of 1:1, combining with position-specific scoring matrix and support vector machine. The best performance achieves an accuracy of 99.98% and AUC 0.9996 in 10-fold cross-validation. The AUC of independent test set reaches 0.9997, which demonstrates the robustness of LipoSVM. The analysis between lysine lipoylation and non-lipoylation fragments shows significant statistical differences. CONCLUSION: A good predictor for lysine lipoylation is built based on position-specific scoring matrix and support vector machine. Meanwhile, an online webserver LipoSVM can be freely downloaded from https://github.com/stars20180811/LipoSVM.