CX3CL1 Action on Microglia Protects from Diet-Induced Obesity by Restoring POMC Neuronal Excitability and Melanocortin System Activity Impaired by High-Fat Diet Feeding

Both hypothalamic microglial inflammation and melanocortin pathway dysfunction contribute to diet-induced obesity (DIO) pathogenesis. Previous studies involving models of altered microglial signaling demonstrate altered DIO susceptibility with corresponding POMC neuron cytological changes, suggesting a link between microglia and the melanocortin system. We addressed this hypothesis using the specific microglial silencing molecule, CX3CL1 (fractalkine), to determine whether reducing hypothalamic microglial activation can restore POMC/melanocortin signaling to protect against DIO. We performed metabolic analyses in high fat diet (HFD)-fed mice with targeted viral overexpression of CX3CL1 in the hypothalamus. Electrophysiologic recording in hypothalamic slices from POMC-MAPT-GFP mice was used to determine the effects of HFD feeding and microglial silencing via minocycline or CX3CL1 on GFP-labeled POMC neurons. Finally, mice with hypothalamic overexpression of CX3CL1 received central treatment with the melanocortin receptor antagonist SHU9119 to determine whether melanocortin signaling is required for the metabolic benefits of CX3CL1. Hypothalamic overexpression of CX3CL1 increased leptin sensitivity and POMC gene expression, while reducing weight gain in animals fed an HFD. In electrophysiological recordings from hypothalamic slice preparations, HFD feeding was associated with reduced POMC neuron excitability and increased amplitude of inhibitory postsynaptic currents. Microglial silencing using minocycline or CX3CL1 treatment reversed these HFD-induced changes in POMC neuron electrophysiologic properties. Correspondingly, blockade of melanocortin receptor signaling in vivo prevented both the acute and chronic reduction in food intake and body weight mediated by CX3CL1. Our results show that suppressing microglial activation during HFD feeding reduces DIO susceptibility via a mechanism involving increased POMC neuron excitability and melanocortin signaling.     

Generalized Approach towards Secretion-Based Protein Production via Neutralization of Secretion-Preventing Cationic Substrate Residues

Many heterologous proteins can be secreted by bacterial ATP-binding cassette (ABC) transporters, provided that they are fused with the C-terminal signal sequence, but some proteins are not secretable even though they carry the right signal sequence. The invention of a method to secrete these non-secretable proteins would be valuable both for understanding the secretory physiology of ABC transporters and for industrial applications. Herein, we postulate that cationic “supercharged” regions within the target substrate protein block the secretion by ABC transporters. We also suggest that the secretion of such substrate proteins can be rescued by neutralizing those cationic supercharged regions via structure-preserving point mutageneses. Surface-protruding, non-structural cationic amino acids within the cationic supercharged regions were replaced by anionic or neutral hydrophilic amino acids, reducing the cationic charge density. The examples of rescued secretions we provide include the spike protein of SARS-CoV-2, glutathione-S-transferase, streptavidin, lipase, tyrosinase, cutinase, growth factors, etc. In summary, our study provides a method to predict the secretability and a tool to rescue the secretion by correcting the secretion-blocking regions, making a significant step in understanding the physiological properties of ABC transporter-dependent protein secretion and laying the foundation for the development of a secretion-based protein-producing platform.     

Col4a3-/- Mice on Balb/C Background Have Less Severe Cardiorespiratory Phenotype and SGLT2 Over-Expression Compared to 129x1/SvJ and C57Bl/6 Backgrounds

Alport syndrome (AS) is a hereditary renal disorder with no etiological therapy. In the preclinical Col4a3^-/- model of AS, disease progression and severity vary depending on mouse strain. The sodium-glucose cotransporter 2 (SGLT2) is emerging as an attractive therapeutic target in cardiac/renal pathologies, but its application to AS remains untested. This study investigates cardiorespiratory function and SGLT2 renal expression in Col4a3^-/- mice from three different genetic backgrounds, 129x1/SvJ, C57Bl/6 and Balb/C. male Col4a3^-/- 129x1/SvJ mice displayed alterations consistent with heart failure with preserved ejection fraction (HFpEF). Female, but not male, C57Bl/6 and Balb/C Col4a3^-/- mice exhibited mild changes in systolic and diastolic function of the heart by echocardiography. Male C57Bl/6 Col4a3^-/- mice presented systolic dysfunction by invasive hemodynamic analysis. All strains except Balb/C males demonstrated alterations in respiratory function. SGLT2 expression was significantly increased in AS compared to WT mice from all strains. However, cardiorespiratory abnormalities and SGLT2 over-expression were significantly less in AS Balb/C mice compared to the other two strains. Systolic blood pressure was significantly elevated only in mutant 129x1/SvJ mice. The results provide further evidence for strain-dependent cardiorespiratory and hypertensive phenotype variations in mouse AS models, corroborated by renal SGLT2 expression, and support ongoing initiatives to develop SGLT2 inhibitors for the treatment of AS.     

The Pro-Inflammatory Deletion Allele of the NF-κB1 Polymorphism Is Characterized by a Depletion of Subunit p50 in Sepsis

The functionally important NF-κB1 promoter polymorphism (−94ins/delATTG) significantly shapes inflammation and impacts the outcome of sepsis. However, exploratory studies elucidating the molecular link of this genotype-dependent pattern are lacking. Accordingly, we analyzed lipopolysaccharide-stimulated peripheral blood mononuclear cells from both healthy volunteers (n = 20) and septic patients (n = 10). All individuals were genotyped for the −94ins/delATTG NF-κB1 promoter polymorphism. We found a diminished nuclear activity of the NF-κB subunit p50 in ID/DD genotypes after 48 h of lipopolysaccharide stimulation compared to II genotypes (p = 0.025). This was associated with higher TNF-α (p = 0.005) and interleukin 6 concentrations (p = 0.014) and an increased production of mitochondrial radical oxygen species in ID/DD genotypes (p = 0.001). Although ID/DD genotypes showed enhanced activation of mitochondrial biogenesis, they still had a significantly diminished cellular ATP content (p = 0.046) and lower mtDNA copy numbers (p = 0.010) compared to II genotypes. Strikingly, these findings were mirrored in peripheral blood mononuclear cells taken from septic patients. Our results emphasize the crucial aspect of considering NF-κB subunits in sepsis. We showed here that the deletion allele of the NF-κB1 (−94ins/delATTG) polymorphism was associated with the lower nuclear activity of subunit p50, which, in turn, was associated with aggravated inflammation and mitochondrial dysfunction.     

Complex and dynamic population structures: synthesis, open questions, and future directions

The population structure of an evolutionary algorithm influences the dissemination and mixing of advantageous alleles, and therefore affects search performance. Much recent attention has focused on the analysis of complex population structures, characterized by heterogeneous connectivity distributions, non-trivial clustering properties, and degree–degree correlations. Here, we synthesize the results of these recent studies, discuss their limitations, and highlight several open questions regarding (1) unsolved theoretical issues and (2) the practical utility of complex population structures for evolutionary search. In addition, we will discuss an alternative complex population structure that is known to significantly influence dynamical processes, but has yet to be explored for evolutionary optimization. We then shift our attention toward dynamic population structures, which have received markedly less attention than their static counterparts. We will discuss the strengths and limitations of extant techniques and present open theoretical and experimental questions and directions for future research. In particular, we will focus on the prospects of “active linking,” wherein edges are dynamically rewired according to the genotypic or phenotypic properties of individuals, or according to the success of prior inter-individual interactions.     

Recurrence of MDD: A prospective study of personality pathology and cognitive distortions.

Major depressive disorder (MDD) is characterized by a high risk of recurrence, especially among individuals whose initial episode occurs during adolescence. Identifying predictors of recurrence of MDD among young samples is therefore of paramount clinical importance. Survival analytic models were used to evaluate the effects of dysfunctional cognitions and Axis II personality pathology on MDD recurrence in a sample of 130 previously, but not currently, depressed young adults. Participants were initially assessed for depression, dysfunctional attitudes, and personality pathology during their first semester in college and then reevaluated via the Longitudinal Interval Follow-up Evaluation interview every 6 months for 18 months. Baseline level of depressive symptoms significantly (HR-1.07, p = .002) predicted recurrence of MDD. In the survival analyses with baseline level of depression serving as a current mood state covariate, overall personality pathology (HR-1.04, p DSM–IV personality disorder cluster scores uniquely predicted recurrence. We discussed the theoretical, empirical, and clinical implications of these findings, and we noted the limitations of the study. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Extension and source/drain design for high-performance FinFET devices

Double gate devices based upon the FinFET architecture are fabricated, with gate lengths as small as 30 nm. Particular attention is given to minimizing the parasitic series resistance. Angled extension implants and selective silicon epitaxy are investigated as methods for minimizing parasitic resistance in FinFETs. Using these two techniques high performance devices are fabricated with on-currents comparable to fully optimized bulk silicon technologies. The influence of fin thickness on device resistance and short channel effects is discussed in detail. Devices are fabricated with fins oriented in the <100> and <100> directions showing different transport properties.     

噪声与低噪声设计的探讨（上篇）

噪声是许多信号处理系统的基本制约因素.同样,它是许多电子设计,特别是接口电路的主要制约条件.在测试与测量、医学成像和高速数据通信等方面的行业趋向都需要越来越高的信息密度.与此同时,半导体工艺的进步能实现更高的数据处理速度和功能密度,但却要降低工作电源电压,由此降低信号幅度.结果是,加大了系统设计对模拟前端噪声性能进行管理的压力.