The oncofetal protein IMP3: a novel grading tool and predictor of poor clinical outcome in human gliomas

Morphologic criteria illustrated in WHO guidelines are the most significant prognostic factor in human gliomas, but novel biomarkers are needed to identify patients with a poorer outcome. The present study examined the expression of the oncofetal protein IMP3 in a series of 135 patients affected by high-grade (grade III and IV) gliomas, correlating the results with proliferative activity, molecular parameters, and clinical and follow-up data. Overall, IMP3 expression was higher in glioblastomas (68%) than in grade III tumors (20%, P < 0.0001), and IMP3-positive high-grade gliomas showed a shorter overall and disease-free survival than negative ones (P = 0.0002 and P = 0.006, resp.). IMP3 expression was significantly associated with the absence of mutations of IDH1 gene (P = 0.0001) and with the unmethylated phenotype of MGMT in high-grade gliomas (P = 0.004). High Ki67 levels were correlated with better prognosis in glioblastomas but IMP3 expression was not correlated with the proliferation index. These findings confirm the role of IMP3 as a marker of poor outcome, also in consideration of its association with IDH1 wild-type phenotype and MGMT unmethylated status. The data suggest that IMP3 staining could identify a subgroup of patients with poor prognosis and at risk of recurrence in high-grade gliomas

Specific V-ATPase expression sub-classifies IDHwt lower-grade gliomas and impacts glioma growth in vivo

Background: Cancer cells use specific V-ATPase subunits to activate oncogenic pathways. Therefore, we investigated V-ATPase deregulation in aggressive gliomas and associated signaling. Methods: V-ATPase genes expression and associated pathways were analyzed in different series of glioma available from public databases, as well as in patients\u2019 cohort. Activation of pathways was analyzed at gene and protein expression levels. A genetic model of glioma in Drosophila melanogaster and mice with GBM patients-derived orthotopic xenografts were used as in vivo models of disease. Findings: GBM and recurrent gliomas display a specific V-ATPase signature. Such signature resolves the heterogeneous class of IDH-wild type lower-grade gliomas, identifying the patients with worse prognosis independently from clinical and molecular features (p = 0\ub703, by Cox proportional-hazards model). In vivo, V-ATPase subunits deregulation significantly impacts tumor growth and proliferation. At the molecular level, GBM-like V-ATPase expression correlates with upregulation of Homeobox genes. Interpretation: Our data identify a V-ATPase signature that accompanies glioma aggressiveness and suggest new entry points for glioma stratification and follow-up. Fund: This work was supported by Fondazione Cariplo (2014\u20131148 to VV), Fondazione IRCCS Ca\u2019 Granda, and Fondazione INGM Grant in Molecular Medicine 2014 (to VV)

Testing an expanded set of sustainable forest management indicators in Mediterranean coppice area

Although coppice forests represent a significant part of the European forest area, especially across southern Countries, they received little attention within the Sustainable Forest Management (SFM) processes and scenarios, whose guidelines have been mainly designed to high forests and national scale. In order to obtain “tailored” information on the degree of sustainability of coppices on the scale of the stand, we evaluated (i) whether the main coppice management options result in different responses of the SFM indicators, and (ii) the degree to which the considered SFM indicators were appropriate in their application at stand level. The study considered three different management options (Traditional Coppice TC, coppice under Natural Evolution NE, and coppice under Conversion to high forest by means of periodical thinning CO). In each of the 43 plots considered in the study, which covered three different European Forest Types, we applied a set of eighteen “consolidated” SFM indicators, covering all the six SFM Criteria (FOREST EUROPE, 2020) and, additionally, tested other sixteen novel indicators shaped for agamic forests and/or applicable at stand level. Results confirmed that several consolidated indicators related to resources status (Growing stock and Carbon stock), health (Defoliation and Forest damage), and socio-economic functions (Net revenue, Energy and Accessibility) were highly appropriate for evaluating the sustainability of coppice at stand level. In addition, some novel indicators related to resources status (Total above ground tree biomass), health (Stand growth) and protective functions (Overstorey cover and Understorey cover) proved to be highly appropriate and able to support the information obtained by the consolidated ones. As a consequence, a subset of consolidated SFM indicators, complemented with the most appropriate novel ones, may represent a valid option to support the evaluation of coppice sustainability at stand level. An integrated analysis of the SFM indicators showed that NE and CO display significant higher environmental performances as compared with TC. In addition, CO has positive effects also on socio-economic issues, while TC -which is an important cultural heritage and a silvicultural option that may help to keep local communities engaged in forestry – combines high wood harvesting rates with dense understory cover. Overall, each of the three management options showed specific sustainability values; as a consequence, their coexistence at a local scale and in accordance with the specific environmental conditions and the social-economic context, is greatly recommended since it may fulfill a wider array of sustainability issue

A GBM-like V-ATPase signature directs cell-cell tumor signaling and reprogramming via large oncosomes

Background: The V-ATPase proton pump controls acidification of intra and extra-cellular milieu in both physiological and pathological conditions. We previously showed that some V-ATPase subunits are enriched in glioma stem cells and in patients with poor survival. In this study, we investigated how expression of a GBM-like V-ATPase pump influences the non-neoplastic brain microenvironment. Methods: Large oncosome (LO) vesicles were isolated from primary glioblastoma (GBM) neurospheres, or from patient sera, and co-cultured with primary neoplastic or non-neoplastic brain cells. LO transcript and protein contents were analyzed by qPCR, immunoblotting and immunogold staining. Activation of pathways in recipient cells was determined at gene and protein expression levels. V-ATPase activity was impaired by Bafilomycin A1 or gene silencing. Findings: GBM neurospheres influence their non-neoplastic microenvironment by delivering the V-ATPase subunit V1G1 and the homeobox genes HOXA7, HOXA10, and POU3F2 to recipient cells via LO. LOs reprogram recipient cells to proliferate, grow as spheres and to migrate. Moreover, LOs are particularly abundant in the circulation of GBM patients with short survival time. Finally, impairment of V-ATPase reduces LOs activity. Interpretation: We identified a novel mechanism adopted by glioma stem cells to promote disease progression via LO-mediated reprogramming of their microenvironment. Our data provide preliminary evidence for future development of LO-based liquid biopsies and suggest a novel potential strategy to contrast glioma progression. Fund: This work was supported by Fondazione Cariplo (2014-1148 to VV) and by the Italian Minister of Health-Ricerca Corrente program 2017 (to SF)

Testing an expanded set of sustainable forest management indicators in Mediterranean coppice area

Although coppice forests represent a significant part of the European forest area, especially across southern Countries, they received little attention within the Sustainable Forest Management (SFM) processes and scenarios, whose guidelines have been mainly designed to high forests and national scale. In order to obtain “tailored” information on the degree of sustainability of coppices on the scale of the stand, we evaluated (i) whether the main coppice management options result in different responses of the SFM indicators, and (ii) the degree to which the considered SFM indicators were appropriate in their application at stand level. The study considered three different management options (Traditional Coppice TC, coppice under Natural Evolution NE, and coppice under Conversion to high forest by means of periodical thinning CO). In each of the 43 plots considered in the study, which covered three different European Forest Types, we applied a set of eighteen “consolidated” SFM indicators, covering all the six SFM Criteria (FOREST EUROPE, 2020) and, additionally, tested other sixteen novel indicators shaped for agamic forests and/or applicable at stand level. Results confirmed that several consolidated indicators related to resources status (Growing stock and Carbon stock), health (Defoliation and Forest damage), and socio-economic functions (Net revenue, Energy and Accessibility) were highly appropriate for evaluating the sustainability of coppice at stand level. In addition, some novel indicators related to resources status (Total above ground tree biomass), health (Stand growth) and protective functions (Overstorey cover and Understorey cover) proved to be highly appropriate and able to support the information obtained by the consolidated ones. As a consequence, a subset of consolidated SFM indicators, complemented with the most appropriate novel ones, may represent a valid option to support the evaluation of coppice sustainability at stand level. An integrated analysis of the SFM indicators showed that NE and CO display significant higher environmental performances as compared with TC. In addition, CO has positive effects also on socio-economic issues, while TC -which is an important cultural heritage and a silvicultural option that may help to keep local communities engaged in forestry – combines high wood harvesting rates with dense understory cover. Overall, each of the three management options showed specific sustainability values; as a consequence, their coexistence at a local scale and in accordance with the specific environmental conditions and the social-economic context, is greatly recommended since it may fulfill a wider array of sustainability issues

The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

The vacuolar H+ ATPase (V-ATPase) is a proton pump responsible for acidification of cellular microenvironments, an activity exploited by tumors to survive, proliferate and resist to therapy. Despite few observations, the role of V-ATPase in human tumorigenesis remains unclear.We investigated the expression of ATP6V0C, ATP6V0A2, encoding two subunits belonging to the V-ATPase V0 sector and ATP6V1C, ATP6V1G1, ATPT6V1G2, ATP6V1G3, which are part of the V1 sector, in series of adult gliomas and in cancer stem cell-enriched neurospheres isolated from glioblastoma (GBM) patients. ATP6V1G1 expression resulted significantly upregulated in tissues of patients with GBM and correlated with shorter patients' overall survival independent of clinical variables.ATP6V1G1 knockdown in GBM neurospheres hampered sphere-forming ability, induced cell death, and decreased matrix invasion, a phenotype not observed in GBM monolayer cultures. Treating GBM organotypic cultures or neurospheres with the selective V-ATPase inhibitor bafilomycin A1 reproduced the effects of ATP6V1G1 siRNA and strongly suppressed expression of the stem cell markers Nestin, CD133 and transcription factors SALL2 and POU3F2 in neurospheres.These data point to ATP6V1G1 as a novel marker of poor prognosis in GBM patients and identify V-ATPase inhibition as an innovative therapeutic strategy for GBM

The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

The vacuolar H+ ATPase (V-ATPase) is a proton pump responsible for acidification of cellular microenvironments, an activity exploited by tumors to survive, proliferate and resist to therapy. Despite few observations, the role of V-ATPase in human tumorigenesis remains unclear.We investigated the expression of ATP6V0C, ATP6V0A2, encoding two subunits belonging to the V-ATPase V0 sector and ATP6V1C, ATP6V1G1, ATPT6V1G2, ATP6V1G3, which are part of the V1 sector, in series of adult gliomas and in cancer stem cell-enriched neurospheres isolated from glioblastoma (GBM) patients. ATP6V1G1 expression resulted significantly upregulated in tissues of patients with GBM and correlated with shorter patients' overall survival independent of clinical variables.ATP6V1G1 knockdown in GBM neurospheres hampered sphere-forming ability, induced cell death, and decreased matrix invasion, a phenotype not observed in GBM monolayer cultures. Treating GBM organotypic cultures or neurospheres with the selective V-ATPase inhibitor bafilomycin A1 reproduced the effects of ATP6V1G1 siRNA and strongly suppressed expression of the stem cell markers Nestin, CD133 and transcription factors SALL2 and POU3F2 in neurospheres.These data point to ATP6V1G1 as a novel marker of poor prognosis in GBM patients and identify V-ATPase inhibition as an innovative therapeutic strategy for GBM

Global Perspectives on Task Shifting and Task Sharing in Neurosurgery.

BACKGROUND: Neurosurgical task shifting and task sharing (TS/S), delegating clinical care to non-neurosurgeons, is ongoing in many hospital systems in which neurosurgeons are scarce. Although TS/S can increase access to treatment, it remains highly controversial. This survey investigated perceptions of neurosurgical TS/S to elucidate whether it is a permissible temporary solution to the global workforce deficit. METHODS: The survey was distributed to a convenience sample of individuals providing neurosurgical care. A digital survey link was distributed through electronic mailing lists of continental neurosurgical societies and various collectives, conference announcements, and social media platforms (July 2018-January 2019). Data were analyzed by descriptive statistics and univariate regression of Likert Scale scores. RESULTS: Survey respondents represented 105 of 194 World Health Organization member countries (54.1%; 391 respondents, 162 from high-income countries and 229 from low- and middle-income countries [LMICs]). The most agreed on statement was that task sharing is preferred to task shifting. There was broad consensus that both task shifting and task sharing should require competency-based evaluation, standardized training endorsed by governing organizations, and maintenance of certification. When perspectives were stratified by income class, LMICs were significantly more likely to agree that task shifting is professionally disruptive to traditional training, task sharing should be a priority where human resources are scarce, and to call for additional TS/S regulation, such as certification and formal consultation with a neurosurgeon (in person or electronic/telemedicine). CONCLUSIONS: Both LMIC and high-income countries agreed that task sharing should be prioritized over task shifting and that additional recommendations and regulations could enhance care. These data invite future discussions on policy and training programs

(1)H MRI of pneumococcal pneumonia in a murine model.

Abstract: Purpose: To detect and quantify pulmonary lesions due to pneumococcal pneumonia in a murine model by H-1 MRI. Materials and Methods: Pneumonia was induced in mice (N = 5) by intranasal administration of about 1 X 10(6) colony-forming units (CFU) of Streptococcus pneumonie. A group of noninfected animals (N = 5) was used as a control group. MRI was performed, 48 hours after infection induction, at 4.7 T. ECG-gated gradient-echo (GRE) sequences with TE = 5 msec were used. After MRI examination, the animals were sacrificed for histological examination. Results: Lungs appeared at MRI as regions with signal intensity (SI) at the level of the noise. Lesions appeared as hyperintense regions over the background and were localized mainly in the apical part of the lungs, in the medial and peribronchial regions. The anatomical localization of the lesions was confirmed by histology. The total lesion volume quantified by MRI data correlated with the total lesion volume quantified by histology. Conclusion: This work shows that standard H-1 MRI allows detection and quantification of lesions due to pneumocoecal pneumonia in mice

Comparison between MRI, microbiology and histology in evaluation of antibiotics in a murine model of thigh infection.

Although in vivo magnetic resonance imaging (MRI) is rapidly becoming a recognised tool in experimental pharmacological research, at the best of our knowledge, scarce application in the field of antibacterial drug research has been reported so far. In this last field, animal models of bacterial infections are used to test the efficacy of novel compounds. In this paper we have explored the potential usefulness of MRI in monitoring the chronological evolution of experimental bacterial infections and the effect of different therapeutic treatments. A murine model of thigh infection induced by Staphylococcus aureus has been used and the efficacy of vancomycin and imipenem/cilastatin has been tested. Three groups of infected animals were studied by microbiology, histology and MRI methods. The results obtained show that in vivo MRI data are highly consistent with microbiological and histological data, allowing, similarly to these commonly used techniques, the efficacy of different antibacterial treatments to be quantified. Our findings suggest that MRI could be used to assess the efficacy of new chemical entities in antibacterial pharmacological research. The advantages of MRI, as a non invasive technique, in comparison with commonly used microbiological and histological methods are discussed