Comparison of three ELISA kits for the differentiation of foot-and-mouth disease virus-infected from vaccinated animals

A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals. Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits, Ceditest® FMDV-NS (Ceditest® kit), UBI® FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI® kit) and a FMDV 3ABC-I-ELISA kit developed at the Lanzhou Veterinary Research Institute. The test parameters (sensitivity and specificity) of the three kits were determined, and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits. The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest® kits was 98.05%, and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI® kits was 94.4%; the sensitivity of both Ceditest® and FMDV 3ABC-I-ELISA kit was 100%. However, the sensitivity of the UBI® kit was only 81.8%. With sera from naive or vaccinated non-infected animals, the specificity of all tests exceeded 90%.     

Quantitative Metaproteomics and Activity-based Protein Profiling of Patient Fecal Microbiome Identifies Host and Microbial Serine-type Endopeptidase Activity Associated With Ulcerative Colitis

The gut microbiota plays an important yet incompletely understood role in the induction and propagation of ulcerative colitis (UC). Organism-level efforts to identify UC-associated microbes have revealed the importance of community structure, but less is known about the molecular effectors of disease. We performed 16S rRNA gene sequencing in parallel with label-free data-dependent LC-MS/MS proteomics to characterize the stool microbiomes of healthy (n = 8) and UC (n = 10) patients. Comparisons of taxonomic composition between techniques revealed major differences in community structure partially attributable to the additional detection of host, fungal, viral, and food peptides by metaproteomics. Differential expression analysis of metaproteomic data identified 176 significantly enriched protein groups between healthy and UC patients. Gene ontology analysis revealed several enriched functions with serine-type endopeptidase activity overrepresented in UC patients. Using a biotinylated fluorophosphonate probe and streptavidin-based enrichment, we show that serine endopeptidases are active in patient fecal samples and that additional putative serine hydrolases are detectable by this approach compared with unenriched profiling. Finally, as metaproteomic databases expand, they are expected to asymptotically approach completeness. Using ComPIL and de novo peptide sequencing, we estimate the size of the probable peptide space unidentified (“dark peptidome”) by our large database approach to establish a rough benchmark for database sufficiency. Despite high variability inherent in patient samples, our analysis yielded a catalog of differentially enriched proteins between healthy and UC fecal proteomes. This catalog provides a clinically relevant jumping-off point for further molecular-level studies aimed at identifying the microbial underpinnings of UC.     

Brain Cytochrome Oxidase in Alzheimer''s Disease

A recent demonstration of markedly reduced (-50%) activity of cytochrome oxidase (CO; complex 4), the terminal enzyme of the mitochondrial enzyme transport chain, in platelets of patients with Alzheimer's disease (AD) suggested the possibility of a systemic and etiologically fundamental CO defect in AD. To determine whether a CO deficiency occurs in AD brain, we measured the activity of CO in homogenates of autopsied brain regions of 19 patients with AD and 30 controls matched with respect to age, postmortem time, sex, and, as indices of agonal status, brain pH and lactic acid concentration. Mean CO activity in AD brain was reduced in frontal (-26%: p less than 0.01), temporal (-17%; p less than 0.05), and parietal (-16%; not significant, p = 0.055) cortices. In occipital cortex and putamen, mean CO levels were normal, whereas in hippocampus, CO activity, on average, was nonsignificantly elevated (20%). The reduction of CO activity, which is tightly coupled to neuronal metabolic activity, could be explained by hypofunction of neurons, neuronal or mitochondrial loss, or possibly by a more primary, but region-specific, defect in the enzyme itself. The absence of a CO activity reduction in all of the examined brain areas does not support the notion of a generalized brain CO abnormality. Although the functional significance of a 16-26% cerebral cortical CO deficit in human brain is not known, a deficiency of this key energy-metabolizing enzyme could reduce energy stores and thereby contribute to the brain dysfunction and neurodegenerative processes in AD.     

Secretases as therapeutic targets for Alzheimer's disease

Accumulation of amyloid-β (Aβ) is widely accepted as the key instigator of Alzheimer’s disease (AD). The proposed mechanism is that accumulation of Aβ results in inflammatory responses, oxidative damages, neurofibrillary tangles and, subsequently, neuronal/synaptic dysfunction and neuronal loss. Given the critical role of Aβ in the disease process, the proteases that produce this peptide are obvious targets. The goal would be to develop drugs that can inhibit the activity of these targets. Protease inhibitors have proved very effective for treating other disorders such as AIDS and hypertension. Mutations in APP (amyloid-β precursor protein), which flanks the Aβ sequence, cause early-onset familial AD, and evidence has pointed to the APP-to-Aβ conversion as a possible therapeutic target. Therapies aimed at modifying Aβ-related processes aim higher up the cascade and are therefore more likely to be able to alter the progression of the disease. However, it is not yet fully known whether the increases in Aβ levels are merely a result of earlier events that were already causing the disease.     

The modification of cortical reorganization and chronic pain by sensory feedback

Recent neuroscientific evidence has revealed that the adult brain is capable of substantial plastic change in areas such as the primary somatosensory cortex that were formerly thought to be modifiable only during early experience. We discuss research on phantom limb pain as well as chronic back pain that revealed functional reorganization in both the somatosensory and the motor system in these chronic pain states. In phantom limb pain patients, cortical reorganization is correlated with the amount of phantom limb pain; in low back pain patients the amount of reorganizational change increases with chronicity. We present a model of the development of chronic pain that assumes an important role of somatosensory pain memories. In phantom limb pain, we propose that those patients who experienced intense pain prior to the amputation will later likely develop enhanced cortical reorganization and phantom limb pain. We show that cortical plasticity related to chronic pain can be reduced by behavioral interventions that provide feedback to the brain areas that were altered by somatosensory pain memories.     

The molecular basis of spinocerebellar ataxia type 48 caused by a de novo mutation in the ubiquitin ligase CHIP

The spinocerebellar ataxias (SCAs) are a class of incurable diseases characterized by degeneration of the cerebellum that results in movement disorder. Recently, a new heritable form of SCA, spinocerebellar ataxia type 48 (SCA48), was attributed to dominant mutations in STIP1 homology and U box-containing 1 (STUB1); however, little is known about how these mutations cause SCA48. STUB1 encodes for the protein C terminus of Hsc70 interacting protein (CHIP), an E3 ubiquitin ligase. CHIP is known to regulate proteostasis by recruiting chaperones via a N-terminal tetratricopeptide repeat domain and recruiting E2 ubiquitin-conjugating enzymes via a C-terminal U-box domain. These interactions allow CHIP to mediate the ubiquitination of chaperone-bound, misfolded proteins to promote their degradation via the proteasome. Here we have identified a novel, de novo mutation in STUB1 in a patient with SCA48 encoding for an A52G point mutation in the tetratricopeptide repeat domain of CHIP. Utilizing an array of biophysical, biochemical, and cellular assays, we demonstrate that the CHIP^A52G point mutant retains E3-ligase activity but has decreased affinity for chaperones. We further show that this mutant decreases cellular fitness in response to certain cellular stressors and induces neurodegeneration in a transgenic Caenorhabditis elegans model of SCA48. Together, our data identify the A52G mutant as a cause of SCA48 and provide molecular insight into how mutations in STUB1 cause SCA48.     

低心血管疾病风险选择性环氧合酶-2抑制剂的虚拟筛选

摘要 目的：寻找具有血栓素A2受体（Thromboxane A2 receptor,TP）抑制作用的选择性环氧合酶-2（Cyclooxygenase-2,COX-2）抑制剂,以期降低其心血管疾病风险。方法：本研究从公开数据库中获取了512种TP抑制剂,通过分子对接、分子动力学模拟和ADMET预测,筛选出化合物TP84。结果：分子对接结果显示,与先前获批的选择性COX-2抑制剂罗非昔布相比,TP84对COX-2的亲和力更高,对环氧合酶-1（Cyclooxygenase-1,COX-1）的亲和力更低;分子动力学模拟进一步表明,模拟过程中TP84与COX-1的结合不稳定,而TP84能稳定结合COX-2,与COX-2的结合自由能是COX-1的3倍;此外,根据ADMET预测,TP84的药物化学、吸收、分布、代谢、排泄和毒性处于类药物候选物的可接受范围内。结论：TP84是一种潜在的低心血管疾病风险选择性COX-2抑制剂。     

慢性阻塞性肺疾病不同表型患者CAT评分与肺功能及预后的相关性分析

摘要 目的：探讨慢性阻塞性肺疾病（chronic obstructive pulmonary disease,COPD）不同表型评估测试问卷（COPD assessment test,CAT）评分与肺功能及预后的关系。方法：收集361例COPD患者临床资料、CAT评分、肺功能检查结果及肺外合并症、肺内并发症等情况,按临床表型分为肺气肿组（n=200）和支气管炎组（n=161）,分析肺气肿组200例和支气管炎组161例COPD患者CAT评分与肺功能及预后的关系。结果：肺气肿组CAT评分高于支气管炎组（P<0.05）,一秒用力呼气容积（FEV₁）占预计值百分比（FEV₁%）、FEV₁/用力肺活量（FVC）低于支气管炎组（P<0.05）,吸气分数（IC/TLC）低于支气管炎组,残总比（RV/TLC）高于支气管炎组（P<0.05）;肺气肿组肺间质性病变、肺动脉高压发生率均高于支气管炎组（P<0.05）;支气管炎、肺气肿组CAT评分均与FEV₁%、FEV₁/FVC、IC/TLC呈负相关（P<0.05）,与RV/TLC呈正相关（P<0.05）,肺气肿各参数相关度更高（P<0.05）;肺气肿组不同CAT评分患者肺间质性病变、肺动脉高压发生率比较差异有统计学意义（P<0.05）,支气管炎组不同CAT评分肺动脉高压发生率比较差异有统计学意义（P<0.05）,随CAT评分的升高,肺气肿组肺间质性病变、肺动脉高压发生率上升,支气管炎组肺动脉高压发生率上升。结论： COPD肺气肿表型CAT评分较支气管炎表型高,肺功能降低更明显,呈现肺过度通气,气流受限特点,更易并发肺间质纤维化、肺动脉高压,且与CAT评分变化密切相关。     

不同程度慢性阻塞性肺疾病患者血清FIB、RDW、MDW及IL-6的表达水平及其与肺功能、急性发作的相关性分析

摘要 目的：分析不同程度慢性阻塞性肺疾病（COPD）患者血清纤维蛋白原（FIB）、红细胞分布宽度（RDW）、单核细胞体积分布宽度（MDW）及白介素6（IL-6）的表达水平及其与肺功能、急性发作的相关性。方法：选择我院自2021年4月至2023年4月接诊的148例COPD患者纳入观察组,根据气流受限严重程度分为轻-中度气流受限组（96例）和重-极重度气流受限组（52例）;另选同期的148例健康体检者纳入对照组。检测所有入选者血清FIB、RDW、MDW及IL-6的表达水平,比较不同组别之间血清FIB、RDW、MDW及IL-6的表达水平,使用Pearson相关性分析血清FIB、RDW、MDW及IL-6与肺功能指标[第一秒用力呼气容积与用力肺活量（FEV₁/FVC）占预计值百分比和第一秒用力呼气容积占预计值百分比（FEV₁%）]的关系,ROC曲线分析血清FIB、RDW、MDW及IL-6对急性发作的预测效能。结果：观察组血清FIB、RDW、MDW及IL-6的表达水平均高于对照组（P<0.05）;重-极重度气流受限组血清FIB、RDW、MDW及IL-6的表达水平均高于轻-中度气流受限组（P<0.05）;经Pearson相关性分析,COPD患者血清FIB、RDW、MDW及IL-6的表达水平与FEV₁/FVC占预计值百分比、FEV₁%均呈负相关（P<0.05）;经ROC曲线分析,血清FIB、RDW、MDW联合IL-6预测COPD急性发作的敏感度为66.35%,特异度为90.42%、AUC为0.912。结论：不同程度COPD患者血清FIB、RDW、MDW及IL-6的表达水平存在差异,血清FIB、RDW、MDW及IL-6与其肺功能密切相关,联合对急性发作具有一定预测价值。     

Dopamine depletion and subsequent treatment with L-DOPA, but not the long-lived dopamine agonist pergolide, enhances activity of the Akt pathway in the rat striatum

Dysregulation of signaling pathways is believed to contribute to Parkinson's disease pathology and l-DOPA-induced motor complications. Long-lived dopamine (DA) agonists are less likely to cause motor complications by virtue of continuous stimulation of DA receptors. In this study, we compared the effects of the unilateral 6-hydroxydopamine lesion and subsequent treatment with l-DOPA and DA agonist pergolide on signaling pathways in rats. Pergolide caused less pronounced behavioral sensitization than l-DOPA (25 mg/kg, i.p., 10 days), particularly at lower dose (0.5 and 0.25 mg/kg, i.p.). Pergolide, but not l-DOPA, reversed lesion-induced up-regulation of preproenkephalin and did not up-regulate preprodynorphine or DA D3 receptor in the lesioned hemisphere. Pergolide was as effective as l-DOPA in reversing the lesion-induced elevation of ERK2 phosphorylation in response to acute apomorphine administration (0.05 mg/kg, s.c.). Chronic l-DOPA significantly elevated the level of Akt phosphorylation at both Thr(308) and Ser(473) and concentration of phosphorylated GSK3alpha, whereas pergolide suppressed the lesion- and/or challenge-induced supersensitive Akt responses. The data indicate that l-DOPA, unlike pergolide, exacerbates imbalances in the Akt pathway caused by the loss of DA. The results support the hypothesis that the Akt pathway is involved in long-term actions of l-DOPA and may be linked to l-DOPA-induced dyskinesia.