Deglycosylated RBD produced in Pichia pastoris as a low-cost sera COVID-19 diagnosis tool and a vaccine candidate.

During the COVID-19 outbreak, numerous tools including protein-based vaccines have been developed. The methylotrophic yeast Pichia pastoris (synonymous to Komagataella phaffii) is an eukaryotic cost-effective and scalable system for recombinant protein production, with the advantages of an efficient secretion system and the protein folding assistance of the secretory pathway of eukaryotic cells. In a previous work, we compared the expression of SARS-CoV-2 Spike Receptor Binding Domain in P. pastoris with that in human cells. Although the size and glycosylation pattern was different between them, their protein structural and conformational features were indistinguishable. Nevertheless, since high mannose glycan extensions in proteins expressed by yeast may be the cause of a nonspecific immune recognition, we deglycosylated RBD in native conditions. This resulted in a highly pure, homogenous, properly folded and monomeric stable protein. This was confirmed by circular dichroism and tryptophan fluorescence spectra and by SEC-HPLC, which were similar to those of RBD proteins produced in yeast or human cells. Deglycosylated RBD was obtained at high yields in a single step, and it was efficient in distinguishing between SARS-CoV-2-negative and positive sera from patients. Moreover, when the deglycosylated variant was used as an immunogen, it elicited a humoral immune response ten times greater than the glycosylated form, producing antibodies with enhanced neutralizing power and eliciting a more robust cellular response. The proposed approach may be used to produce at a low cost, many antigens that require glycosylation to fold and express, but do not require glycans for recognition purposes.

RUNX1 Is Regulated by Androgen Receptor to Promote Cancer Stem Markers and Chemotherapy Resistance in Triple Negative Breast Cancer.

Triple negative breast cancer (TNBC) is an aggressive breast cancer subtype for which no effective targeted therapies are available. Growing evidence suggests that chemotherapy-resistant cancer cells with stem-like properties (CSC) may repopulate the tumor. The androgen receptor (AR) is expressed in up to 50% of TNBCs, and AR inhibition decreases CSC and tumor initiation. Runt-related transcription factor 1 (RUNX1) correlates with poor prognosis in TNBC and is regulated by the AR in prostate cancer. Our group has shown that RUNX1 promotes TNBC cell migration and regulates tumor gene expression. We hypothesized that RUNX1 is regulated by the AR and that both may work together in TNBC CSC to promote disease recurrence following chemotherapy. Chromatin immunoprecipitation sequencing (ChIP-seq) experiments in MDA-MB-453 revealed AR binding to RUNX1 regulatory regions. RUNX1 expression is upregulated by dihydrotestosterone (DHT) in MDA-MB-453 and in an AR+-TNBC HCI-009 patient-derived xenograft (PDX) tumors (p < 0.05). RUNX1 is increased in a CSC-like experimental model in MDA-MB-453 and SUM-159PT cells (p < 0.05). Inhibition of RUNX1 transcriptional activity reduced the expression of CSC markers. Interestingly, RUNX1 inhibition reduced cell viability and enhanced paclitaxel and enzalutamide sensitivity. Targeting RUNX1 may be an attractive strategy to potentiate the anti-tumor effects of AR inhibition, specifically in the slow-growing CSC-like populations that resist chemotherapy which lead to metastatic disease.

Effect of Background Music on Attentional Control in Older and Young Adults.

Healthy aging may be accompanied by cognitive decline that includes diminished attentional control, an executive function that allows us to focus our attention while inhibiting distractors. Previous studies have demonstrated that background music can enhance some executive functions in both young and older adults. According to the Arousal-Mood Theory, the beneficial influence of background music on cognitive performance would be related to its ability to increase the arousal level of the listeners and to improve their mood. Consequently, stimulating and pleasant music might enhance attentional control. Therefore, the aims of this study were (1) to determine if the influence of background music, and more specifically its arousal level, might improve attentional control in older adults and (2) whether this effect is similar across older and young adults. Older and young adults performed a visuo-spatial flanker task during three auditory conditions: stimulating music, relaxing music, and silence. Participants had to indicate as fast and as accurately as possible the direction of a central arrow, which was flanked by congruent or incongruent arrows. As expected, reaction times were slower for the incongruent compared to congruent trials. Interestingly, this difference was significantly greater under the relaxing music condition compared to other auditory conditions. This effect was the same across both age groups. In conclusion, relaxing music seems to interfere with visuo-spatial attentional control compared to stimulating music and silence, regardless of age.

Influence of Background Musical Emotions on Attention in Congenital Amusia.

Congenital amusia in its most common form is a disorder characterized by a musical pitch processing deficit. Although pitch is involved in conveying emotion in music, the implications for pitch deficits on musical emotion judgements is still under debate. Relatedly, both limited and spared musical emotion recognition was reported in amusia in conditions where emotion cues were not determined by musical mode or dissonance. Additionally, assumed links between musical abilities and visuo-spatial attention processes need further investigation in congenital amusics. Hence, we here test to what extent musical emotions can influence attentional performance. Fifteen congenital amusic adults and fifteen healthy controls matched for age and education were assessed in three attentional conditions: executive control (distractor inhibition), alerting, and orienting (spatial shift) while music expressing either joy, tenderness, sadness, or tension was presented. Visual target detection was in the normal range for both accuracy and response times in the amusic relative to the control participants. Moreover, in both groups, music exposure produced facilitating effects on selective attention that appeared to be driven by the arousal dimension of musical emotional content, with faster correct target detection during joyful compared to sad music. These findings corroborate the idea that pitch processing deficits related to congenital amusia do not impede other cognitive domains, particularly visual attention. Furthermore, our study uncovers an intact influence of music and its emotional content on the attentional abilities of amusic individuals. The results highlight the domain-selectivity of the pitch disorder in congenital amusia, which largely spares the development of visual attention and affective systems.

Age-related changes in attention control and their relationship with gait performance in older adults with high risk of falls.

BACKGROUND: Falls are the leading cause of injury-related deaths in the elderly worldwide. Both gait impairment and cognitive decline have been shown to constitute major fall risk factors. However, further investigations are required to establish a more precise link between the influence of age on brain systems mediating executive cognitive functions and their relationship with gait disturbances, and thus help define novel markers and better guide remediation strategies to prevent falls. METHODS: Event-related functional magnetic resonance imaging (fMRI) was used to evaluate age-related effects on the recruitment of executive control brain network in selective attention task, as measured with a flanker paradigm. Brain activation patterns were compared between twenty young (21 years ±â€¯2.5) and thirty-four old participants (72 years ±â€¯5.3) with high fall risks. We then determined to what extend age-related differences in activation patterns were associated with alterations in several gait parameters, measured with electronic devices providing a precise quantitative evaluation of gait, as well as with alterations in several aspects of cognitive and physical abilities. RESULTS: We found that both young and old participants recruited a distributed fronto-parietal-occipital network during interference by incongruent distractors in the flanker task. However, additional activations were observed in posterior parieto-occipital areas in the older relative to the younger participants. Furthermore, a differential recruitment of both the left dorsal parieto-occipital sulcus and precuneus was significantly correlated with higher gait variability. Besides, decreased activation in the right cerebellum was found in the older with poorer cognitive processing speed scores. CONCLUSIONS: Overall results converge to indicate greater sensitivity to attention interference and heightened recruitment of cortical executive control systems in the elderly with fall risks. Critically, this change was associated with selective increases in gait variability indices, linking attentional control with gait performance in elderly with high risks of falls.

Brain networks mediating the influence of background music on selective attention.

Prevalent across societies and times, music has the ability to enhance attention, a property relevant to clinical applications, but the underlying brain mechanisms remain unknown. It is also unclear whether music produces similar or differential effects with advancing age. Here, we used event-related functional magnetic resonance imaging to investigate the influence of music exposure evoking four types of emotions on distinct attentional components measured with a modified attention network test, across 19 young (21 ± 2.6) and 33 old participants (72 ± 5.4). We then determined whether music-related effects differed across age groups and whether they were associated with particular acoustic features. Background music during selective attention requiring distractor conflict resolution was associated with faster response times and greater activations of fronto-parietal areas during happy and high-arousing music, whereas sad and low-valence music was associated with slower responses and greater occipital recruitment. Shifting and altering components of attention were unaffected. The influence of music on performance and brain networks was similar between age groups. These behavioral and neuroimaging results demonstrate the importance of affective music dimensions, particularly arousal, in enhancing selective attention processes. This study adds novel support to the benefits of music in the rehabilitation of attention functions.