Abstract
Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more relevant information ...and more predictive data for in vivo tests. Willing to increase the knowledge about Medulloblastoma Brain tumor we are developing Human cerebellar organoids derived from human iPSC: three-dimensional cultures that include multiple stem cell types, cell organization and features of human cerebellum. Notably, Medulloblastoma is the most common brain tumor affecting infants and stands as a cause for a high percentage of morbidity and mortality among cancer patients. To identify new Medulloblastoma driver genes we set up an innovative way to modify human cerebellar organoid, developing the first in-vitro 3D model of Medulloblastoma. We are mimicking human pathological conditions modifying this organoids with different putative oncogenes. Notably thanks to this type of model, we are able to study the functions of new Medulloblastoma driver genes in modulating stem cells differentiation and commitment in a three-dimensional structure, during the development of the organoid and the tumor. We speculate that our new Medulloblastoma 3D culture systems hold great promise for applications in infant tumor research, cancer cell biology and drug discovery, being the first human 3D in-vitro model that resembles human pathological conditions.
The dysbiosis of the oral microbiome is associated with both localized and systemic diseases. Modulating the resident microbial communities by the dietary consumption of probiotics has become an ...appealing means to promote host health by either restoring host-microbe balance or preventing dysbiosis. Most probiotics strategies target the intestinal microbiome, but little is known about their impact on the oral microbiome. We analyzed here the saliva microbiome from 21 volunteers, longitudinally collected before, during, and after consumption of a commercial probiotic and a standard yoghurt using 16S amplicon sequencing. The alpha diversity of the saliva microbiome had a statistically significant increase (P-value = 0.0011) in one of the groups that consumed the probiotic. The overall structure of the microbiome was however not significantly impacted by the probiotic, although oligotyping analysis revealed that both Streptococci and Lactobacilli present in the probiotic product persisted in the saliva microbiome. In contrast, non-probiotic yoghurt consumption had a lesser impact on the overall diversity and Lactobacillus and Streptococcus persistence. Our results suggest that consumption of commercial probiotics in healthy subjects increase the overall diversity of the oral cavity microbiome in the short term, but such dietary interventions are not able to substantially modify the structure of the microbiome.
Notch1 pathway activation is required for group 3 medulloblastoma initiation.
The identity of the cell of origin is a key determinant of cancer subtype, progression, and prognosis. Group 3 ...medulloblastoma (MB) is a malignant childhood brain cancer with poor prognosis and few candidates as putative cell of origin. We overexpressed the group 3 MB genetic drivers
MYC
and
Gfi1
in different candidate cells of origin in the postnatal mouse cerebellum. We found that S100b
+
cells are competent to initiate group 3 MB, and we observed that S100b
+
cells have higher levels of Notch1 pathway activity compared to Math1
+
cells. We found that additional activation of Notch1 in Math1
+
and Sox2
+
cells was sufficient to induce group 3 MB upon
MYC
/
Gfi1
expression. Together, our data suggest that the Notch1 pathway plays a critical role in group 3 MB initiation.
Medulloblastoma (MB) is the most common pediatric brain tumor with high morbidity and mortality. Among the four major MB groups, patients with Group3 MB currently have the worst outcome and nearly ...50% are metastatic at the time of diagnosis. However, the molecular mechanisms underlying Group 3 MB are still unknown. Exploiting in vivo transfection of mouse cerebellar cells, we tested different patient- specific combinations of genes for their ability to induce Group 3 MB in mice. We identified Otx2 and cMYC as strong inducers of tumors faithfully recapitulating human Group 3 MB.;
To identify a druggable signaling pathway, we analyzed sequencing data of human patients harboring Otx2/cMYC overexpression. Among all the putative oncosuppressors, SMARCA4 is the most frequently mutated. Indeed, co-expression of SMARCA4 with Otx2 and cMYC is able to block tumorigenesis, through regulation of CDKN2B and CRABP1 expression. Interestingly, patient-specific SMARCA4T910M mutant is able to block wild- type SMARCA4 effects in a dominant-negative manner. Since SMARCA4 has an antagonistic relationship with histone methyltransferase EZH2, we used EZH2-inhibitors to recapitulate SMARCA4 antitumorigenic effects.;
Little is known about Group 3 MB developmental origin and several embryonic mouse cerebellar progenitors have been proposed as possible cell of origin. Here, we found that in postnatal mouse cerebellum, the S100b+ cells are competent to induce medulloblastoma while Math1, Sox2 and Ascl1 positive cells seem to be unresponsive to oncogenic insults. Taken together, our data suggest that the competency of different cells to develop MB could change during brain development, depending on their developmental stage and cellular identity.
•Massive sampling of Chianti red wines, which are distinguished with a PDO label.•Major, trace and Rare Earth elements were used as fingerprints for Chianti identification.•Multivariate ...classification is able to discriminate Chianti and non-Chianti samples.
Chianti is a precious red wine and enjoys a high reputation for its high quality in the world wine market. Despite this, the production region is small and product needs efficient tools to protect its brands and prevent adulterations. In this sense, ICP-MS combined with chemometrics has demonstrated its usefulness in food authentication. In this study, Chianti/Chianti Classico, authentic wines from vineyard of Toscana region (Italy), together samples from 18 different geographical regions, were analyzed with the objective of differentiate them from other Italian wines. Partial Least Squares-Discriminant Analysis (PLS-DA) identified variables to discriminate wine geographical origin. Rare Earth Elements (REE), major and trace elements all contributed to the discrimination of Chianti samples. General model was not suited to distinguish PDO red wines from samples, with similar chemical fingerprints, collected in some regions. Specific classification models enhanced the capability of discrimination, emphasizing the discriminant role of some elements.
Heterozygous mutations in the gene encoding RagD GTPase were shown to cause a novel autosomal dominant condition characterized by kidney tubulopathy and cardiomyopathy. We previously demonstrated ...that RagD, and its paralogue RagC, mediate a non-canonical mTORC1 signaling pathway that inhibits the activity of TFEB and TFE3, transcription factors of the MiT/TFE family and master regulators of lysosomal biogenesis and autophagy. Here we show that RagD mutations causing kidney tubulopathy and cardiomyopathy are "auto- activating", even in the absence of Folliculin, the GAP responsible for RagC/D activation, and cause constitutive phosphorylation of TFEB and TFE3 by mTORC1, without affecting the phosphorylation of "canonical" mTORC1 substrates, such as S6K. By using HeLa and HK-2 cell lines, human induced pluripotent stem cell-derived cardiomyocytes and patient-derived primary fibroblasts, we show that RRAGD auto-activating mutations lead to inhibition of TFEB and TFE3 nuclear translocation and transcriptional activity, which impairs the response to lysosomal and mitochondrial injury. These data suggest that inhibition of MiT/TFE factors plays a key role in kidney tubulopathy and cardiomyopathy syndrome.
To evaluate whether an unexpected poor response (cases with ≤3 oocytes) leads to a reduction in the pregnancy rate in IVF cycles compared to a suboptimal response (controls with 4-9 oocytes) in women ...with adequate ovarian reserve.
A nested case-control study performed in a retrospective cohort of couples undergoing IVF at the Infertility Unit of the ASST Lariana. Cases and controls had adequate ovarian reserve and were matched 1:1 for female age and number of previous cycles. Cumulative clinical pregnancy rate per oocyte retrieval was the main outcome.
Overall, 113 cases and 113 matched controls were included; the median number of available oocytes was 2 and 6, respectively. The cumulative pregnancy rate per cycle was significantly reduced in cases compared to controls with a crude odds ratio = 0.45 95% Confidence Interval: 0.28-0.82. A binomial logistic model indicated that an increase in one oocyte increases the odds for cumulative pregnancy rate per cycle by 1.27 in women with 9 oocytes or less. The cumulative pregnancy rates per cycle in cases and controls, according to female age were respectively: 29% versus 54% in patients aged <35 years (p = 0.036); 22% versus 43% in patients aged 36-39 years (p = 0.048) and 11% versus 13% in patients 40-45 years old (p = 0.72). Patients belonging to older age groups showed decreasing probability of cumulative clinical pregnancy rates both among cases and controls group (p < 0.05).
The number of available oocytes significantly affects the probability of success in IVF cycles with unexpected impaired ovarian response.
Tuna “Bottarga” is an artisanal food speciality prepared from the roe of Bluefin Tuna (Thunnus thynnus). Due to the recent restriction of Tuna catches fixed by EU, cheaper substitutes from yellowfin ...tuna (Thunnus albacares) are currently available in the mass distribution. Since the product is sold sliced or grated, morphological distinctive features may be lost, and fraudulent substitution is possible. In this study we tested DNA mini-barcoding and infrared spectroscopy as tools to trace its authenticity. A certified chemical standard, fresh (n = 4) and processed (n = 11) ovarian identified by the morphological observation of the tuna catches, commercial bottargasamples (n = 5) and artificial aged aliquots of all these samples (n = 50) were used in the experiments. The results showed that infrared spectroscopy failed in the taxonomic distinction, whereas enabled PLS-DA classification of aged samples (and thus the identification of fresh prepared artisanal products) with model non-error rate in fitting equal to 0.96, and 0.89 in cross-validation. Conversely, DNA mini-barcoding with mtDNA control region, success in species identification (100% maximum identity by the standard comparison BLAST approach against GenBank), regardless the processing state of the product.
•Fraudolent substitution of seafood products is possible.•A combined molecular and spectroscopic approach was applied as tool to authenticity control of Bluefin tuna roe.•DNA mini-barcoding provided the taxonomical authentication of the products, regardless the processing.•PCA analysis applied to Infrared spectra enable a discrimination based on storage condition.
The uneven worldwide vaccination coverage against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and emergence of variants escaping immunity call for broadly effective and easily ...deployable therapeutic agents. We have previously described the human single-chain scFv76 antibody, which recognizes SARS-CoV-2 Alpha, Beta, Gamma and Delta variants. We now show that scFv76 also neutralizes the infectivity and fusogenic activity of the Omicron BA.1 and BA.2 variants. Cryoelectron microscopy (cryo-EM) analysis reveals that scFv76 binds to a well-conserved SARS-CoV-2 spike epitope, providing the structural basis for its broad-spectrum activity. We demonstrate that nebulized scFv76 has therapeutic efficacy in a severe hACE2 transgenic mouse model of coronavirus disease 2019 (COVID-19) pneumonia, as shown by body weight and pulmonary viral load data. Counteraction of infection correlates with inhibition of lung inflammation, as observed by histopathology and expression of inflammatory cytokines and chemokines. Biomarkers of pulmonary endothelial damage were also significantly reduced in scFv76-treated mice. The results support use of nebulized scFv76 for COVID-19 induced by any SARS-CoV-2 variants that have emerged so far.
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Milazzo et al. demonstrate that the nebulized scFv76 antibody has therapeutic efficacy in a severe hACE2-transgenic mouse model of COVID-19 pneumonia. In addition to previous proof of activity against SARS-CoV-2 Alpha, Beta, Gamma, and Delta, they show that scFv76 can neutralize Omicron BA.1 and BA.2 by recognizing a well-conserved spike epitope modeled by cryo-EM.
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