Comparative biodistribution in mice of cyanine dyes loaded in lipid nanoparticles

Two near infrared cyanine dyes, DiD (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine perchlorate) and ICG (Indocyanine Green) were loaded in lipid nanoparticles (LNP). DiD-LNP and ICG-LNP presented similar physicochemical characteristics (hydrodynamic diameter, polydispersity, zeta potential), encapsulation efficiency, and colloidal stability when stored in PBS buffer. However, whereas DiD had similar biodistribution than cholesteryl-1-¹⁴C-oleate ([¹⁴C]CHO, a constituent of the nanoparticle used as a reference radiotracer), ICG displayed a different biodistribution pattern, similar to that of the free dye, indicative of its immediate leakage from the nanovector after blood injection. NMR spectroscopy using Proton NOE (Nuclear Overhauser Effect) measurements showed that the localization of the dye in the lipid nanoparticles was slightly different: ICG, more amphiphilic than DiD, was found both inside the lipid core and at particle interface, whereas DiD, more hydrophobic, appeared exclusively located inside the particle core. The ICG release rate from the particles was 7% per 1 month under storage conditions (4 °C, dark, 10% of lipids), whereas no leakage could be detected for DiD. ICG leakage increased considerably in the presence of BSA 40 g/L (45% leakage in 24 h at 100 mg/mL of lipids), because of the high affinity of the fluorophore for plasma proteins. On the contrary, no DiD leakage was observed, until high dilution of the nanoparticles which triggered their dissociation (45% leakage in 24 h at 1 mg/mL of lipids). Altogether, the subtle difference in dye localization into the nanoparticles, the partial dissociation of the LNP in diluted media, and more importantly the high ICG affinity for plasma proteins, accounted for the differences observed in the fluorescence biodistribution after tail vein injection of the dye-loaded nanoparticles.     

低频小探头超声内镜对胰腺内分泌肿瘤术前定位诊断的价值

目的 评价低频小探头超声内镜检查(LFMPS)在胰腺内分泌肿瘤术前定位诊断中的临床价值。方法 2000年6月至2002年6月期间21例临床拟诊为胰腺内分泌肿瘤的患者术前行腹部 B超、螺旋 CT、磁共振(MRI)及 LFMPS探查(Fujinon 7.5 MHz低频小探头超声及超声系统),检查结果与外科术中定位和病理结果对照,评估LFMPS对胰腺内分泌肿瘤的术前定位诊断价值。结果21例患者中17例经外科手术及术后病理证实为内分泌肿瘤,4例未手术。其中胰岛素瘤16例(头部9例、体部 3例、尾部4例),胰腺外血管活性肠肽瘤1例,检出病灶的平均直径 2.02 cm。LFMPS确诊14例(82.4％),B超确诊9例(52.9％),螺旋CT确诊15例(88.2％),MRI确诊12例(70.6％),其中LFMPS对位于胰腺头、体部肿瘤以及直径<1cm病灶的确诊率优于其他常规影像方法。结论LFMPS对胰腺内分泌肿瘤的术前定位诊断准确率较高,且与肿瘤的位置与大小有关。     

Ser9 phosphorylation causes cytoplasmic detention of I2/SET in Alzheimer disease

The nuclear protein I₂^PP2A/SET, an endogenous inhibitor of protein phosphatase-2A (PP2A), is increased and translocated to the cytoplasm in the neurons of Alzheimer's disease (AD) brains, and PP2A activity in cytoplasm is compromised. However, it is not fully understood how SET is retained in the cytoplasm. By generating a phosphorylation site-specific antibody, we found in the present study that SET is phosphorylated at Ser9, by which it is accumulated in the cytoplasm of the AD brains. Further studies demonstrate that both the phosphor-mimic and casein kinase (CK)II-mediated phosphorylation at Ser9 interferes with the formation of the SET/importin-α/importin-β complex, and thus inhibits SET nuclear import and induces the cytoplasmic detention of SET. Interestingly, Ser9 is nested in the center of the sequence ⁶AKVSKK¹¹ of SET, which is consistent with a classical nuclear localization signal (NLS). To test whether ⁶AKVSKK¹¹ is a new NLS of SET, we mutated SET lysine 7, lysine 10, and lysine 11 to alanine acid (K7A, K10A, K11A) respectively, and expressed these mutants in HEK293/tau cells. We found that expression of SET (K11A) led to a nuclear import defect of SET, and application of a synthesized peptide Tat-AAKVSKKE that can competitively bind to importin α/β resulted in cytoplasmic detention of SET. Finally, phosphorylation of SET aggravates PP2A inhibition and leads to tau hyperphosphorylation. In conclusion, the current study has identified a novel mechanism that causes cytoplasmic detention of SET with a new NLS-dependent CKII-associated phosphorylation of Ser9, suggesting that inhibition of CKII arrests cytoplasmic accumulation of SET and thus preserves PP2A activity in AD brains.     

Variability and effect sizes of intracranial current source density estimations during pain: Systematic review,experimental findings,and future perspectives

Pain arises from the integration of sensory and cognitive processes in the brain, resulting in specific patterns of neural oscillations that can be characterized by measuring electrical brain activity. Current source density (CSD) estimation from low‐resolution brain electromagnetic tomography (LORETA) and its standardized (sLORETA) and exact (eLORETA) variants, is a common approach to identify the spatiotemporal dynamics of the brain sources in physiological and pathological pain‐related conditions. However, there is no consensus on the magnitude and variability of clinically or experimentally relevant effects for CSD estimations. Here, we systematically examined reports of sample size calculations and effect size estimations in all studies that included the keywords pain, and LORETA, sLORETA, or eLORETA in Scopus and PubMed. We also assessed the reliability of LORETA CSD estimations during non‐painful and painful conditions to estimate hypothetical sample sizes for future experiments using CSD estimations. We found that none of the studies included in the systematic review reported sample size calculations, and less than 20% reported measures of central tendency and dispersion, which are necessary to estimate effect sizes. Based on these data and our experimental results, we determined that sample sizes commonly used in pain studies using CSD estimations are suitable to detect medium and large effect sizes in crossover designs and only large effects in parallel designs. These results provide a comprehensive summary of the effect sizes observed using LORETA in pain research, and this information can be used by clinicians and researchers to improve settings and designs of future pain studies.     

A distributed network supports spatiotemporal cerebral dynamics of visual naming

ObjectiveCerebral spatiotemporal dynamics of visual naming were investigated in epilepsy patients undergoing stereo-electroencephalography (SEEG) monitoring.MethodsBrain networks were defined by Parcel-Activation-Resection-Symptom matching (PARS) approach by matching high-gamma (50–150 Hz) modulations (HGM) in neuroanatomic parcels during visual naming, with neuropsychological outcomes after resection/ablation of those parcels. Brain parcels with >50% electrode contacts simultaneously showing significant HGM were aligned, to delineate spatiotemporal course of naming-related HGM.ResultsIn 41 epilepsy patients, neuroanatomic parcels showed sequential yet temporally overlapping HGM course during visual naming. From bilateral occipital lobes, HGM became increasingly left lateralized, coursing through limbic system. Bilateral superior temporal HGM was noted around response time, and right frontal HGM thereafter. Correlations between resected/ablated parcels, and post-surgical neuropsychological outcomes showed specific regional groupings.ConclusionsConvergence of data from spatiotemporal course of HGM during visual naming, and functional role of specific parcels inferred from neuropsychological deficits after resection/ablation of those parcels, support a model with six cognitive subcomponents of visual naming having overlapping temporal profiles.SignificanceCerebral substrates supporting visual naming are bilaterally distributed with relative hemispheric contribution dependent on cognitive demands at a specific time. PARS approach can be extended to study other cognitive and functional brain networks.     

Fast oscillations >40 Hz localize the epileptogenic zone: An electrical source imaging study using high-density electroencephalography

ObjectiveFast Oscillations (FO) >40 Hz are a promising biomarker of the epileptogenic zone (EZ). Evidence using scalp electroencephalography (EEG) remains scarce. We assessed if electrical source imaging of FO using 256-channel high-density EEG (HD-EEG) is useful for EZ identification.MethodsWe analyzed HD-EEG recordings of 10 focal drug-resistant epilepsy patients with seizure-free postsurgical outcome. We marked FO candidate events at the time of epileptic spikes and verified them by screening for an isolated peak in the time-frequency plot. We performed electrical source imaging of spikes and FO within the Maximum Entropy of the Mean framework. Source localization maps were validated against the surgical cavity.ResultsWe identified FO in five out of 10 patients who had a superficial or intermediate deep generator. The maximum of the FO maps was localized inside the cavity in all patients (100%). Analysis with a reduced electrode coverage using the 10–10 and 10–20 system showed a decreased localization accuracy of 60% and 40% respectively.ConclusionsFO recorded with HD-EEG localize the EZ. HD-EEG is better suited to detect and localize FO than conventional EEG approaches.SignificanceThis study acts as proof-of-concept that FO localization using 256-channel HD-EEG is a viable marker of the EZ.     

基于CT的髌骨下极骨折分型

目的 基于三维CT图像对髌骨下极骨折分型。 方法 回顾性分析本院2018年5月至2020年11月67例髌骨下极骨折的三维CT图像并分型。用Kappa系数评估观察者间可信度和观察者自身可信度。 结果 髌骨下极骨折分为无移位骨折（Ⅰ型）和移位骨折（Ⅱ型）两型。移位骨折可分为3个亚型：Ⅱa型,大块型骨折,包括髌骨下极1枚孤立性骨折块,或2枚较大的骨折块;Ⅱb型,粉碎性骨折,多枚较小的骨折块,包括冠状位和矢状位骨折块;Ⅱc型,袖套样撕脱骨折,沿髌骨下极整个弧面的呈内外方向的长弧形骨折,多为粉碎性的,骨折块非常薄。本研究中Ⅰ型8例,Ⅱa型11例,Ⅱb型42例,Ⅱc型6例。观察者之间的可信度平均K值为0.782（0.682 ~ 0.896）,基本可信;观察者自身可信度平均K值为0.837（0.786 ~ 0.884）,完全可信。 结论 本文髌骨下极骨折分型方法有较高的可信度及可重复性,对临床有一定的指导意义。     

Di-lysine motif-like sequences formed by deleting the C-terminal domain of aquaporin-4 prevent its trafficking to the plasma membrane

Aquaporin-4 is a transmembrane water channel protein, the C-terminal domain of which is facing the cytosol. In the process of investigating the role of the C-terminal domain of aquaporin-4 with regard to intracellular trafficking, we observed that a derivative of aquaporin-4, in which the C-terminal 53 amino acids had been removed (Δ271-323), was localized to intracellular compartments, including the endoplasmic reticulum, but was not expressed on the plasma membranes. This was determined by immunofluorescence staining and labeling of the cells with monoclonal antibody specifically recognizing the extracellular domain of aquaporin-4, followed by confocal microscopy and flow cytometry. Deletion of additional amino acids in the C-terminal domain of aquaporin-4 led to its redistribution to the plasma membrane. This suggests that the effect of the 53-amino acid deletion on the subcellular localization of aquaporin-4 could be attributed to the formation of a signal at the C terminus that retained aquaporin-4 in intracellular compartments, rather than the loss of a signal required for plasma membrane targeting. Substitution of the lysine at position 268 with alanine could rescue the Δ271-323-associated retention in the cytosol, suggesting that the C-terminal sequence of the mutant served as a signal similar to a di-lysine motif.     

Whole Exome Sequencing Reveals Uncommon Mutations in the Recently Identified Fanconi Anemia Gene SLX4/FANCP

Fanconi anemia (FA) is a rare genetic disorder characterized by congenital malformations, progressive bone marrow failure (BMF), and susceptibility to malignancies. FA is caused by biallelic or hemizygous mutations in one of 15 known FA genes, whose products are involved in the FA/BRCA DNA damage response pathway. Here, we report on a patient with previously unknown mutations of the most recently identified FA gene, SLX4/FANCP. Whole exome sequencing (WES) revealed a nonsense mutation and an unusual splice site mutation resulting in the partial replacement of exonic with intronic bases, thereby removing a nuclear localization signal. Immunoblotting detected no residual SLX4 protein, which was consistent with abrogated interactions with XPF/ERCC1 and MUS81/EME1. This cellular finding did not result in a more severe clinical phenotype than that of previously reported FA‐P patients. Our study additionally exemplifies the versatility of WES for the detection of mutations in heterogenic disorders such as FA.     

Primary angiosarcoma of the breast: Diagnosis with computer-assisted MRI-guided radio-guided occult lesion localization (ROLL) technique

Primary angiosarcoma of the breast is a rare type of breast cancer that represents approximately 0.04% of all primary breast tumors. We report herein a case of primary breast angiosarcoma that was only visible on magnetic resonance imaging (MRI) examination. The patient presented with a palpable right breast lump that was not visible either on ultrasonography and mammography. MRI showed a lesion of the right breast that presented washout kinetics. MRI-guided biopsy allowed histopathological examination of the tumor that was further confirmed as primary angiosarcoma. Subsequently, MRI guided ROLL (radio-guided occult lesion localization) technique was used for localizing the lesion prior to surgery.