C9orf72 expansion within astrocytes reduces metabolic flexibility in amyotrophic lateral sclerosis

It is important to understand how the disease process affects the metabolic pathways in amyotrophic lateral sclerosis and whether these pathways can be manipulated to ameliorate disease progression. To analyse the basis of the metabolic defect in amyotrophic lateral sclerosis we used a phenotypic metabolic profiling approach. Using fibroblasts and reprogrammed induced astrocytes from C9orf72 and sporadic amyotrophic lateral sclerosis cases we measured the production rate of reduced nicotinamide adenine dinucleotides (NADH) from 91 potential energy substrates simultaneously. Our screening approach identified that C9orf72 and sporadic amyotrophic lateral sclerosis induced astrocytes have distinct metabolic profiles compared to controls and displayed a loss of metabolic flexibility that was not observed in fibroblast models. This loss of metabolic flexibility, involving defects in adenosine, fructose and glycogen metabolism, as well as disruptions in the membrane transport of mitochondrial specific energy substrates, contributed to increased starvation induced toxicity in C9orf72 induced astrocytes. A reduction in glycogen metabolism was attributed to loss of glycogen phosphorylase and phosphoglucomutase at the protein level in both C9orf72 induced astrocytes and induced neurons. In addition, we found alterations in the levels of fructose metabolism enzymes and a reduction in the methylglyoxal removal enzyme GLO1 in both C9orf72 and sporadic models of disease. Our data show that metabolic flexibility is important in the CNS in times of bioenergetic stress

Systematic bioinformatic analysis of expression profiles of human vestibular sensory epithelia in response to aminoglycoside toxicity

Deafness and balance dysfunction are commonly caused by a loss of sensory hair cells (HCs) due to a combination of factors including genetics, ageing, noise, drugs, infections, noise. Spontaneous HC regeneration occurs in non-mammals, but in mammals, this capacity is limited to the vestibular sensory epithelium, and this decreases over time. Recently, the induction of the transcription factor ATOH1 has been shown to promote the transdifferentiation of human and mouse vestibular supporting cells (SCs) into immature HCs. However, despite these promising results, additional genes and pathways are required for functional HC maturation. This project aimed to further elucidate the mechanism of aminoglycoside toxicity in the human inner ear and to identify novel biomarkers and candidate genes for HCs regeneration. A new RNAseq dataset of purified human vestibular epithelia treated for 24hr with gentamicin was characterised and compared with published datasets, allowing the identification of novel toxic effects, activated signalling pathways and putative genes promoting HC regeneration. Gene expression of the purified human vestibular epithelia showed a global inhibition of the transcription and translation machineries. GJB2 (coding for connexin 26) and endoplasmic reticulum (ER) stress genes were the most altered during the early phases of the insult. These features were exacerbated after total HCs loss, as observed when compared with the published dataset of the whole human utricle treated with gentamicin for 48hr. The analyses of the whole utricle treated with gentamicin identified genes involved in the spontaneous formation of cilia and in cytoskeletal rearrangement. A cross-species comparison with chick sensory epithelia showed significant differences in inflammatory response upon toxic insults and in genes/pathways important for HCs development and regeneration. A deeper analysis of these differences allowed the identification of MYC and LIF as two new potential candidate genes for the promotion of HC regeneration

Focal testicular lesions. Multiparametric us features and association with histopathology

NTRODUCTION AND OBJECTIVES: Testicular cancer represents the most common solid tumors in young men and malignant germ cell tumors constitute the majority of these masses. Orchiectomy is the treatment of choice for intratesticular masses. However, it is important to recognize those benign conditions for which orchiectomy is unwarranted because approximately 5-10% of all testicular masses are postoperatively identified as benign testicular lesions. Ultrasound features of solid scrotal tumors are often non-specific. US, however, identifies the lesion in the largest number of cases. Here, we evaluate the histopatologic features which determine echogenicity, echotexture, stiffness, and vascularity of different testicular masses. METHODS: 98 testicular lesions investigated with colour Doppler US for which histological specimens were available for review were enclosed. 47 lesions had also CEUS, 34 elastography. Imaging features were correlated with histological characteristics. RESULTS: Most testicular cancers were hypervascular at colour Doppler interrogation and with increased consistency at elastography. 6/22 hypovascular lesions with small vessels at colour Doppler interrogation were hypervascular at CEUS. Seminomas presented with lower echogenicity compared to other cancers, mixed tumors were heterogeneous. Irrespective of histotype, abundance of stroma was the main factor determining increased echogenicity. Other factors were hyalization, interleaved necrotic areas and tumor nests, athrophic changes of the surrounding parenchyma. Heterogeous appearance was observed in presence of different histotypes, calcifications, necrotic or fibrotic areas. Lesions were avascular at colour Doppler interrogation if vessels were lacking or were very small. In the latter case, they were vascularized at CEUS. Lesions with abundant stroma were hard at elastography, while necrotic changes caused soft or mixed appearance CONCLUSIONS: The different histological features of testicular lesions determine their sonographic appearance. US is highly sensitive for detection of testicular lesions, but specificity is low.The different US modes, if taken individually, are non-specific but if used together improve lesion characterization. Preoperative imaging and correct characterization of focal testicular lesions play a quite important role in guiding the surgical approach to these patients allowing the identification of testicular lesions amenable to treatment with testicular-sparing surger

Integration of Cellular and Humoral Immune Responses as an Immunomonitoring Tool for SARS-CoV-2 Vaccination in Healthy and Fragile Subjects

Cellular and humoral immunity are both required for SARS-CoV-2 infection recovery and vaccine efficacy. The factors affecting mRNA vaccination-induced immune responses, in healthy and fragile subjects, are still under investigation. Thus, we monitored the vaccine-induced cellular and humoral immunity in healthy subjects and cancer patients after vaccination to define whether a different antibody titer reflected similar rates of cellular immune responses and if cancer has an impact on vaccination efficacy. We found that higher titers of antibodies were associated with a higher probability of positive cellular immunity and that this greater immune response was correlated with an increased number of vaccination side effects. Moreover, active T-cell immunity after vaccination was associated with reduced antibody decay. The vaccine-induced cellular immunity appeared more likely in healthy subjects rather than in cancer patients. Lastly, after boosting, we observed a cellular immune conversion in 20% of subjects, and a strong correlation between pre- and post-boosting IFN-γ levels, while antibody levels did not display a similar association. Finally, our data suggested that integrating humoral and cellular immune responses could allow the identification of SARS-CoV-2 vaccine responders and that T-cell responses seem more stable over time compared to antibodies, especially in cancer patients