Usefulness of Corticomedullary-Phase CT Urography in Patients with Suspected Acute Renal Colic Visiting the Emergency Department.

Purpose: To evaluate the sensitivity of corticomedullary-phase imaging for detecting urinary stones in patients with renal colic who visited the emergency department. Materials and Methods: This retrospective study included 253 patients with suspected renal colic from two tertiary hospitals in South Korea, who visited the emergency department and underwent CT urography. Two radiologists blinded to the clinical history independently reviewed the corticomedullary-phase images. The sensitivity for identifying urinary stones were evaluated for each reviewer. After the initial evaluation, the images were re-evaluated based on patient history. The sensitivity of re-evaluation were recorded. Results: Of 253 patients, 150 (59%) had urinary stones. Among them, significant stones were observed in 138 patients (92%), and obstructive changes on CT in 124 patients (82.7%). For identifying significant urinary stones, the sensitivity was 98.6% (136/138) for both the reviewers. For identifying significant urinary stones with urinary obstruction, the sensitivity was 99.2% (123/124) for reviewer 1, and 100% (124/124) for reviewer 2. The sensitivity for identifying significant stones increased from 98.6% to 100% for reviewer 1, and from 98.6% to 99.3% for reviewer 2 in the re-evaluation session. Conclusion: The corticomedullary-phase CT urography was sensitive for diagnosing urolithiasis in patients with acute renal colic who visited the emergency department.

Learning Correlations between Internal Coordinates to Improve 3D Cartesian Coordinates for Proteins.

We consider a generic representation problem of internal coordinates (bond lengths, valence angles, and dihedral angles) and their transformation to 3-dimensional Cartesian coordinates of a biomolecule. We show that the internal-to-Cartesian process relies on correctly predicting chemically subtle correlations among the internal coordinates themselves, and learning these correlations increases the fidelity of the Cartesian representation. We developed a machine learning algorithm, Int2Cart, to predict bond lengths and bond angles from backbone torsion angles and residue types of a protein, which allows reconstruction of protein structures better than using fixed bond lengths and bond angles or a static library method that relies on backbone torsion angles and residue types in a local environment. The method is able to be used for structure validation, as we show that the agreement between Int2Cart-predicted bond geometries and those from an AlphaFold 2 model can be used to estimate model quality. Additionally, by using Int2Cart to reconstruct an IDP ensemble, we are able to decrease the clash rate during modeling. The Int2Cart algorithm has been implemented as a publicly accessible python package at https://github.com/THGLab/int2cart.

In situ immunogenic clearance induced by a combination of photodynamic therapy and rho-kinase inhibition sensitizes immune checkpoint blockade response to elicit systemic antitumor immunity against intraocular melanoma and its metastasis.

BACKGROUND: Uveal melanoma (UM) is the most frequent intraocular malignancy and is resistant to immunotherapy. Nearly 50% of patients with UM develop metastatic disease, and the overall survival outcome remains very poor. Therefore, a treatment regimen that simultaneously targets primary UM and prevents metastasis is needed. Here, we suggest an immunotherapeutic strategy for UM involving a combination of local photodynamic therapy (PDT), rho-kinase (ROCK) inhibitor, and PD-1/PD-L1 immune checkpoint blockade. METHODS: The antitumor efficacy and immune response of monotreatment or combinational treatment were evaluated in B16F10-bearing syngeneic mouse models. Abscopal antitumor immune responses induced by triple-combinational treatment were validated in syngeneic bilateral B16F10 models. After each treatment, the immune profiles and functional examinations were assessed in tumors and tumor draining lymph nodes by flow cytometry, ELISA, and immunofluorescence assays. In orthotopic intraocular melanoma models, the location of the immune infiltrate in the tumor microenvironment (TME) was evaluated after each treatment by multiplex immunohistochemistry and metastatic nodules were monitored. RESULTS: PDT with Ce6-embedded nanophotosensitizer (FIC-PDT) elicited immunogenic cell death and stimulated antigen-presenting cells. In situ immunogenic clearance induced by a combination of FIC-PDT with ripasudil, a clinically approved ROCK inhibitor, stimulated antigen-presenting cells, which in turn primed tumor-specific cytotoxic T cells. Moreover, local immunogenic clearance sensitized PD-1/PD-L1 immune checkpoint blockade responses to reconstruct the TME immune phenotypes of cold tumors into hot tumors, resulting in recruitment of robust cytotoxic CD8+ T cells in the TME, propagation of systemic antitumor immunity to mediate abscopal effects, and prolonged survival. In an immune-privileged orthotopic intraocular melanoma model, even low-dose FIC-PDT and ripasudil combined with anti-PD-L1 antibody reduced the primary tumor burden and prevented metastasis. CONCLUSIONS: A combination of localized FIC-PDT and a ROCK inhibitor exerted a cancer vaccine-like function. Immunogenic clearance led to the trafficking of CD8+ T cells into the primary tumor site and sensitized the immune checkpoint blockade response to evoke systemic antitumor immunity to inhibit metastasis, one of the major challenges in UM therapy. Thus, immunogenic clearance induced by FIC-PDT and ROCK inhibitor combined with anti-PD-L1 antibody could be a potent immunotherapeutic strategy for UM.

Intra-mitochondrial self-assembly to overcome the intracellular enzymatic degradation of l-peptides.

The design of peptide-based therapeutics is generally based on the replacement of l-amino acids with d-isomers to obtain improved therapeutic efficiency. However, d-isomers are expensive and frequently induce undesirable immune responses. In the present work, we demonstrate that an intra-mitochondrially self-assembling amphiphilic peptide exhibits analogous activity in both d- and l-isomeric forms. This outcome is in contrast to the general observation considering higher therapeutic efficiencies of d-isomers compared with l-analogues. This suggests that l-peptides overcome proteolytic degradation during intra-mitochondrial self-assembly both in vitro and in vivo.

Metabolic profiles and free radical scavenging activity of Cordyceps bassiana fruiting bodies according to developmental stage.

The metabolic profiles of Cordyceps bassiana according to fruiting body developmental stage were investigated using gas chromatography-mass spectrometry. We were able to detect 62 metabolites, including 48 metabolites from 70% methanol extracts and 14 metabolites from 100% n-hexane extracts. These metabolites were classified as alcohols, amino acids, organic acids, phosphoric acids, purine nucleosides and bases, sugars, saturated fatty acids, unsaturated fatty acids, or fatty amides. Significant changes in metabolite levels were found according to developmental stage. Relative levels of amino acids, purine nucleosides, and sugars were higher in development stage 3 than in the other stages. Among the amino acids, valine, isoleucine, lysine, histidine, glutamine, and aspartic acid, which are associated with ABC transporters and aminoacyl-tRNA biosynthesis, also showed higher levels in stage 3 samples. The free radical scavenging activities, which were significantly higher in stage 3 than in the other stages, showed a positive correlation with purine nucleoside metabolites such as adenosine, guanosine, and inosine. These results not only show metabolic profiles, but also suggest the metabolic pathways associated with fruiting body development stages in cultivated C. bassiana.

Effects of light intensity and nitrogen starvation on glycerolipid, glycerophospholipid, and carotenoid composition in Dunaliella tertiolecta culture.

Time-course variation of lipid and carotenoid production under high light (300 µE/m²s) and nitrogen starvation conditions was determined in a Dunaliella tertiolecta strain. Nanoelectrospray (nanoESI) chip based direct infusion was used for lipid analysis and ultra-performance liquid chromatography (UPLC) coupled with a photodiode array (PDA) or atmospheric chemical ionization mass spectrometry (APCI-MS) was used for carotenoid analysis. A total of 29 lipids and 7 carotenoids were detected. Alterations to diacylglyceryltrimethylhomoserine (DGTS) and digalactosyldiacylglycerol (DGDG) species were significant observations under stress conditions. Their role in relation to the regulation of photosynthesis under stress condition is discussed in this study. The total carotenoid content was decreased under stress conditions, while ã-carotene was increased under nitrate-deficient cultivation. The highest productivity of carotenoid was attained under high light and nitrate sufficiency (HLNS) condition, which result from the highest level of biomass under HLNS. When stress was induced at stationary phase, the substantial changes to the lipid composition occurred, and the higher carotenoid content and productivity were exhibited. This is the first report to investigate the variation of lipids, including glycerolipid, glycerophospholipid, and carotenoid in D. tertiolecta in response to stress conditions using lipidomics tools.

Alteration of media composition and light conditions change morphology, metabolic profile, and beauvericin biosynthesis in Cordyceps bassiana mycelium.

Metabolic alterations of Cordyceps bassiana mycelium were investigated under the following culture medium and light conditions: dextrose agar supplemented with 0.5% yeast extract (SDAY) medium with light (SL), SDAY medium without light (SD), nut medium without light (ND), and iron-supplemented SDAY medium without light (FD). The levels of asparagine, aspartic acid, glutamic acid, glutamine, histidine, lysine, ornithine, and proline were significantly higher under SD and SL conditions. The levels of most of the alcohols, saturated fatty acids, unsaturated fatty acids, fatty acid esters, sterols, and terpenes were higher under the ND condition than in the other conditions, but beauvericin was not detectable under the ND condition. The FD condition was favorable for the enhanced production of aminomalonic acid, malic acid, mannonic acid, and erythritol. Thus, the metabolic characteristics of C. bassiana can be manipulated by varying the cultivation conditions, rendering this fungus potentially favorable as a nutraceutical and medicinal resource.

Gas chromatography/mass spectrometry-based metabolic profiling and differentiation of ginseng roots according to cultivation age using variable selection.

Ginseng roots are an important herbal resource worldwide, and the adulteration of ginseng with age is recognized as a serious problem. It is therefore crucial to develop objective criteria or standard protocols for differentiating ginseng root samples according to their cultivation age. The reported study used GC/MS combined with multivariate statistical analysis with variable selection to obtain metabolic profiling and an optimal partial least squares-discriminant analysis (PLS-DA) model for the differentiation of ginseng according to cultivation age. Relative levels of various metabolites, such as amino acids, alcohols, fatty acids, organic acids, and sugars, were measured for various ginseng cultivation ages. Increasing cultivation age resulted in the production of higher levels of panaxynol and panaxydol, which are active polyacetylene compounds in ginseng. In addition, optimized PLS-DA models for the prediction of ginseng age were obtained by selecting variables based on a variable importance in the projection cut-off value of 1.3. Proline, glucaric acid, mannose, gluconic acid, glucuronic acid, myoinositol, panaxydol, and panaxynol are suggested as key and relevant compounds with which to differentiate the age of ginseng samples. The findings of this study suggest that GC/MS-based metabolic profiling can be used to differentiate ginseng samples according to cultivation age.

Electrochemically driven, electrode-addressable formation of functionalized polydopamine films for neural interfaces.

The electrode-specific formation of polydopamine films is achieved by applying positive voltage to the target electrodes at pH 6.0. The functionalization of the films is simultaneously carried out by co-depositing dopamine with molecules of interest onto the electrode.

Retinal ganglion cell responses to voltage and current stimulation in wild-type and rd1 mouse retinas.

Retinal prostheses are being developed to restore vision for those with retinal diseases such as retinitis pigmentosa or age-related macular degeneration. Since neural prostheses depend upon electrical stimulation to control neural activity, optimal stimulation parameters for successful encoding of visual information are one of the most important requirements to enable visual perception. In this paper, we focused on retinal ganglion cell (RGC) responses to different stimulation parameters and compared threshold charge densities in wild-type and rd1 mice. For this purpose, we used in vitro retinal preparations of wild-type and rd1 mice. When the neural network was stimulated with voltage- and current-controlled pulses, RGCs from both wild-type and rd1 mice responded; however the temporal pattern of RGC response is very different. In wild-type RGCs, a single peak within 100 ms appears, while multiple peaks (approximately four peaks) with ∼ 10 Hz rhythm within 400 ms appear in RGCs in the degenerated retina of rd1 mice. We find that an anodic phase-first biphasic voltage-controlled pulse is more efficient for stimulation than a biphasic current-controlled pulse based on lower threshold charge density. The threshold charge densities for activation of RGCs both with voltage- and current-controlled pulses are overall more elevated for the rd1 mouse than the wild-type mouse. Here, we propose the stimulus range for wild-type and rd1 retinas when the optimal modulation of a RGC response is possible.

Combinatorial activin receptor-like kinase/Smad and basic fibroblast growth factor signals stimulate the differentiation of human embryonic stem cells into the cardiac lineage.

The transforming growth factor beta/bone morphogenetic protein-activated Smad signaling pathway plays a complicated role in the maintenance of human embryonic stem cell (hESC) pluripotency and in the cell fate decision of hESCs. Here, we report that sustained inhibition of the transforming growth factor beta type I receptor (also termed activin receptor-like kinase or ALK) using a chemical inhibitor selective for ALK4/5/7 (ALKi) leads to the cardiac differentiation of hESCs under feeder-free and serum-free conditions. Treatment with ALKi reduced Smad2/3 phosphorylation and increased Smad1/5/8 phosphorylation in hESCs, suggesting a requirement for active Smad1/5/8 signaling for cardiac induction in these cells when ALK/Smad2/3 is inhibited. Importantly, active basic fibroblast growth factor (bFGF) signaling was also required for ALKi-mediated cardiac differentiation of monolayer-cultured hESCs. The FGF receptor inhibitor SU5402 blocked ALKi-mediated cardiac induction in hESCs, whereas bone morphogenetic protein-4 enhanced the ALKi-induced increase in phospho-Smad1/5/8 levels but failed to induce the cardiac differentiation of hESCs and instead promoted trophoblastic differentiation. We also confirmed that ALKi potentially enhanced the cardiac differentiation of human embryoid bodies, as determined by expression of cardiac-specific markers, increased beating areas, and action potential recorded from beating areas. These results demonstrate that an ALKi could be used as a potential cardiac-inducing agent and that the development of culture conditions that provide an appropriate balance between ALK/Smad and bFGF signaling is necessary to direct the fate of hESCs into the cardiac lineage.

Effects of clopidogrel on the pharmacokinetics of sibutramine and its active metabolites.

Sibutramine is metabolized by the enzymes CYP2B6 and CYP2C19 into 2 active metabolites, M1 (mono-desmethyl sibutramine) and M2 (di-desmethyl sibutramine). Clopidogrel is a mechanism-based inhibitor of CYP2B6 and CYP2C19. In this study, 13 extensive metabolizers of CYP2B6 and CYP2C19 were evaluated to clarify whether clopidogrel inhibits the formation of the active metabolites of sibutramine. In the control phase, each subject received a 15-mg oral dose of sibutramine. After a washout period of 2 weeks, in the clopidogrel phase, the subjects received 300 mg of clopidogrel on the first day and then 75-mg once daily for 6 days. One hour after the last dosing of clopidogrel, all subjects received 15-mg of sibutramine. Compared with the control phase, the mean sibutramine and M1 plasma concentrations were higher after clopidogrel treatment. Clopidogrel significantly increased the half-life (242% of control phase) and area under the plasma concentration-time curve from 0 to infinity (AUC(inf)) (227% of control phase) of sibutramine and decreased the apparent oral clearance (31% of control phase) of sibutramine. Pharmacokinetic analysis showed significant increases in the AUC(inf) (162% of control phase) of M1. The CYP2B6 and CYP2C19 inhibitor clopidogrel significantly inhibited the formations of M1 from sibutramine and M2 from sibutramine by 37% and 64%, respectively. Therefore, CYP2B6 and CYP2C19 are in vivo catalysts for the formation of the 2 active metabolites of sibutramine.