Detection of Brucellae in peripheral blood mononuclear cells for monitoring therapeutic efficacy of brucellosis infection.

BACKGROUND: Brucellosis is one of the most severe widespread zoonoses caused by the Gram-negative bacterium Brucella species. The diagnosis and clinical assessment of human brucellosis are very important for the management of patients, while there is a lack of effective methods to detect Brucellae. Classical culture of Brucella species is time consuming and often fails. A simple and sensitive assay is needed for diagnosis of Brucella infection and monitoring of treatment in man. METHODS: Blood samples and peripheral blood mononuclear cells (PBMCs) were collected from 154 patients hospitalized for brucellosis. Brucella antibodies were detected by Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (SAT) and enzyme-linked immunosorbent assay (ELISA). Intracellular Brucellae were detected by blood culture and immunofluorescence staining (IFS). RESULTS: Among 154 brucellosis patients, 59.7% (92/154) were antibody reactive by RBPT, 81.8% (126/154) by SAT and 95.5% (147/154) by ELISA, respectively. Only 3.2% (5/154) of patient blood samples resulted in positive Brucella culture, while 68.8% (106/154) carried IFS detectable Brucella antigens in PBMCs. Gender (P = 0.01) but not age (P > 0.05) was a significant risk factor. The frequency of intracellular Brucella antigens was similar between patients receiving different treatment regimens (P > 0.05). However, a significant decrease of intracellular Brucellae was observed only in patients with acute brucellosis after the third course of treatment (P < 0.05), suggesting that current regimens to treat chronic brucellosis were not effective. CONCLUSIONS: IFS appears a sensitive assay for detection of Brucella antigens in PBMCs and could be used for diagnosis and therapeutic monitoring of brucellosis in clinical practice

Structural covariance in subcortical stroke patients measured by automated MRI-based volumetry

A network-level investigation of the volumetric changes of subcortical stroke patients is still lacking. Here, we explored the alterations of structural covariance caused by subcortical stroke with automated brain volumetry. T1-weighed brain MRI scans were obtained from 63 normal controls (NC), 46 stroke patients with infarct in left internal capsule (CI_L), 33 stroke patients with infarct in right internal capsule (CI_R). We performed automatic anatomical segmentation of the T1-weighted brain images with AccuBrain. Volumetric structural covariance analyses were first performed within the basal ganglia structures that were both identified by voxel-based morphometry with AAL atlas and AccuBrain. Subsequently, we additionally included the infratentorial regions that were particularly quantified by AccuBrain for the structural covariance analyses and investigated the alterations of anatomical connections within these subcortical regions in CI_L and CI_R compared with NC. The association between the regional brain volumetry and motor function was also evaluated in stroke groups. There were significant and extensive volumetric differences in stroke patients. These significant regions were generally symmetric for CI_L and CI_R group depending on the side of stroke, involving both regions close to lesions and remote regions. The structural covariance analyses revealed the synergy volume alteration in subcortical regions both in CI_L and CI_R group. In addition, the alterations of volumetric structural covariance were more extensive in CI_L group than CI_R group. Moreover, we found that the subcortical regions with atrophy contributed to the deficits of motor function in CI_R group but not CI_L group, indicating a lesion-side effect of brain volumetric changes after stroke. These findings indicated that the chronic subcortical stroke patients have extensive disordered anatomical connections involving the whole-brain level network, and the connections patterns depend on the lesion-side. Keywords: Brain segmentation, Brain volumetric changes, MRI imaging, Stroke, Structural covariance, Lesion-side effec

Acyl-CoA synthase ACSL4: an essential target in ferroptosis and fatty acid metabolism

Abstract. Long-chain acyl-coenzyme A (CoA) synthase 4 (ACSL4) is an enzyme that esterifies CoA into specific polyunsaturated fatty acids, such as arachidonic acid and adrenic acid. Based on accumulated evidence, the ACSL4-catalyzed biosynthesis of arachidonoyl-CoA contributes to the execution of ferroptosis by triggering phospholipid peroxidation. Ferroptosis is a type of programmed cell death caused by iron-dependent peroxidation of lipids; ACSL4 and glutathione peroxidase 4 positively and negatively regulate ferroptosis, respectively. In addition, ACSL4 is an essential regulator of fatty acid (FA) metabolism. ACSL4 remodels the phospholipid composition of cell membranes, regulates steroidogenesis, and balances eicosanoid biosynthesis. In addition, ACSL4-mediated metabolic reprogramming and antitumor immunity have attracted much attention in cancer biology. Because it facilitates the cross-talk between ferroptosis and FA metabolism, ACSL4 is also a research hotspot in metabolic diseases and ischemia/reperfusion injuries. In this review, we focus on the structure, biological function, and unique role of ASCL4 in various human diseases. Finally, we propose that ACSL4 might be a potential therapeutic target

Characterization of novel Omp31 antigenic epitopes of Brucella melitensis by monoclonal antibodies

Abstract Background Brucellosis is a severe zoonotic disease worldwide. Detection and identification of Brucella species are essential to prevent or treat brucellosis in humans and animals. The outer membrane protein-31 (Omp31) is a major protein of Brucellae except for B. abortus, while the Omp31 antigenic epitopes have not been extensively characterized yet. Results A total of 22 monoclonal antibodies (mAbs) were produced against Omp31 of Brucella (B.) melitensis, of which 13 recognized five linear epitopes, 7 reacted with semi-conformational epitopes and 2 reacted with conformational epitopes, respectively. The mAb isotypes were 11 (50%) IgG2a, 5 (23%) IgG1 and 6 (27%) IgM. On the basis of epitope recognition and reactivity levels, 8 mAbs including 3 IgM and 5 IgG clones were considered as highly reactive and potentially diagnostic antibodies. Among these mAbs, 7A3 (IgG1), 5B1 (IgG2a), 2C1 (IgG2a) and 5B3 (IgG2a) reacted with differently conserved linear epitopes of B. melitensis, B. ovis, B. suis and B. canis strains, while 5H3 (IgG2a) highly reacted with a conformational epitope of Omp31 when tested with several immunoassays. Conclusions These potent monoclonal antibodies can be used for identifying Omp31 antigens or detecting B. melitensis and other Brucella species beyond B. abortus in vitro or in vivo

A Deep Learning Model System for Diagnosis and Management of Adnexal Masses

Appropriate clinical management of adnexal masses requires a detailed diagnosis. We retrospectively collected ultrasound images of 1559 cases from the first Center of Chinese PLA General Hospital and developed a fully automatic deep learning (DL) model system to diagnose adnexal masses. The DL system contained five models: a detector, a mass segmentor, a papillary segmentor, a type classifier, and a pathological subtype classifier. To test the DL system, 462 cases from another two hospitals were recruited. The DL system identified benign, borderline, and malignant tumors with macro-F1 scores that varied from 0.684 to 0.791, a benefit to preventing both delayed and overextensive treatment. The macro-F1 scores of the pathological subtype classifier to categorize the benign masses varied from 0.714 to 0.831. The detailed classification can inform clinicians of the corresponding complications of each pathological subtype of benign tumors. The distinguishment between borderline and malignant tumors and inflammation from other subtypes of benign tumors need further study. The accuracy and sensitivity of the DL system were comparable to that of the expert and intermediate sonographers and exceeded that of the junior sonographer