Heterogeneity of cancer-initiating cells within glioblastoma.

Malignant gliomas, particularly glioblastoma multiforme (GBM), account for the majority of brain tumors. Their incidence is increasing world wide and they are incurable. Although a transient response to therapy is observed, tumor recurrence is inevitable and occurs within tissue that has received cytotoxic therapy. This suggests that a subpopulation of resistant cells is responsible for tumor regrowth. The treatment of GBMs represents a daunting challenge to clinicians due principally to the lack of effective therapeutic options. One explanation for this is the marked cellular and genetic heterogeneity within and across these types of tumors. Unravelling the cellular composition of gliomas and describing cell lineage relationships are essential for therapeutic breakthroughs. The recent proposal that a small percentage of cells with stem cells characteristics are responsible for tumor initiation and growth has sparked an interest in applying approaches used to study somatic stem cells toward an understanding of the cellular elements responsible for cancer progression and recurrence. To outline the relevance of these findings is the purpose of this review

442. LV Expressing MR Reporter Genes Allows In Vivo Monitoring of Stem Cell Gene Therapy

Somatic stem cells (SSC) have raised interest because of their therapeutic potential in both cell-based and gene therapy applications. Towards this goal, tracking the fate of either delivered cells or of genetically modified endogenous cells is of utmost importance. Diverse imaging approaches are available for cell tracking and among these MRI shows a greater resolution and allows direct anatomic correlation and long-term studies of dynamic cell migration on living animals. Superparamagnetic iron oxide (SPIO) has been used to label SSC in vitro and to make them detectable in vivo upon transplantation. However, major limitations of this approach are the progressive dilution of the contrast media among cell progeny and the need for ex vivo SPIO loading. We thus explored an alternative strategy based on the combination of lentiviral vectors (LV), which efficiently transduce SSC both ex vivo and in vivo and allow long-term expression in their progeny, and MR reporter genes, able to increase iron uptake and accumulation into different cell types

Mild Hypoxia Enhances Proliferation and Multipotency of Human Neural Stem Cells

Neural stem cells (NSCs) represent an optimal tool for studies and therapy of neurodegenerative diseases. We recently established a v-myc immortalized human NSC (IhNSC) line, which retains stem properties comparable to parental cells. Oxygen concentration is one of the most crucial environmental conditions for cell proliferation and differentiation both in vitro and in vivo. In the central nervous system, physiological concentrations of oxygen range from 0.55 to 8% oxygen. In particular, in the in the subventricular zone niche area, it's estimated to be 2.5 to 3%.We investigated in vitro the effects of 1, 2.5, 5, and 20% oxygen concentrations on IhNSCs both during proliferation and differentiation. The highest proliferation rate, evaluated through neurosphere formation assay, was obtained at 2.5 and 5% oxygen, while 1% oxygen was most noxious for cell survival. The differentiation assays showed that the percentages of β-tubIII+ or MAP2+ neuronal cells and of GalC+ oligodendrocytes were significantly higher at 2.5% compared with 1, 5, or 20% oxygen at 17 days in vitro. Mild hypoxia (2.5 to 5% oxygen) promoted differentiation into neuro-oligodendroglial progenitors as revealed by the higher percentage of MAP2+/Ki67+ and GalC+/Ki67+ residual proliferating progenitors, and enhanced the yield of GABAergic and slightly of glutamatergic neurons compared to 1% and 20% oxygen where a significant percentage of GFAP+/nestin+ cells were still present at 17 days of differentiation.These findings raise the possibility that reduced oxygen levels occurring in neuronal disorders like cerebral ischemia transiently lead to NSC remaining in a state of quiescence. Conversely, mild hypoxia favors NSC proliferation and neuronal and oligodendroglial differentiation, thus providing an important advance and a useful tool for NSC-mediated therapy of ischemic stroke and neurodegenerative diseases like Parkinson's disease, multiple sclerosis, and Alzheimer's disease

A novel promising laccase from the psychrotolerant and halotolerant Antarctic marine Halomonas sp. M68 strain

Microbial communities inhabiting the Antarctic Ocean show psychrophilic and halophilic adaptations conferring interesting properties to the enzymes they produce, which could be exploited in biotechnology and bioremediation processes. Use of cold- and salt-tolerant enzymes allows to limit costs, reduce contaminations, and minimize pretreatment steps. Here, we report on the screening of 186 morphologically diverse microorganisms isolated from marine biofilms and water samples collected in Terra Nova Bay (Ross Sea, Antarctica) for the identification of new laccase activities. After primary screening, 13.4 and 10.8% of the isolates were identified for the ability to oxidize 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and the dye azure B, respectively. Amongst them, the marine Halomonas sp. strain M68 showed the highest activity. Production of its laccase-like activity increased six-fold when copper was added to culture medium. Enzymatic activity-guided separation coupled with mass spectrometry identified this intracellular laccase-like protein (named Ant laccase) as belonging to the copper resistance system multicopper oxidase family. Ant laccase oxidized ABTS and 2,6-dimethoxy phenol, working better at acidic pHs The enzyme showed a good thermostability, with optimal temperature in the 40–50°C range and maintaining more than 40% of its maximal activity even at 10°C. Furthermore, Ant laccase was salt- and organic solvent-tolerant, paving the way for its use in harsh conditions. To our knowledge, this is the first report concerning the characterization of a thermo- and halo-tolerant laccase isolated from a marine Antarctic bacterium

Functional Characterization of Two Variants at the Intron 6-Exon 7 Boundary of the KCNQ2 Potassium Channel Gene Causing Distinct Epileptic Phenotypes

Pathogenic variants in KCNQ2 encoding for Kv7.2 potassium channel subunits have been found in patients affected by widely diverging epileptic phenotypes, ranging from Self-Limiting Familial Neonatal Epilepsy (SLFNE) to severe Developmental and Epileptic Encephalopathy (DEE). Thus, understanding the pathogenic molecular mechanisms of KCNQ2 variants and their correlation with clinical phenotypes has a relevant impact on the clinical management of these patients. In the present study, the genetic, biochemical, and functional effects prompted by two variants, each found in a non-familial SLNE or a DEE patient but both affecting nucleotides at the KCNQ2 intron 6-exon 7 boundary, have been investigated to test whether and how they affected the splicing process and to clarify whether such mechanism might play a pathogenetic role in these patients. Analysis of KCNQ2 mRNA splicing in patient-derived lymphoblasts revealed that the SLNE-causing intronic variant (c.928-1G > C) impeded the use of the natural splice site, but lead to a 10-aa Kv7.2 in frame deletion (Kv7.2 p.G310Δ10); by contrast, the DEE-causing exonic variant (c.928G > A) only had subtle effects on the splicing process at this site, thus leading to the synthesis of a full-length subunit carrying the G310S missense variant (Kv7.2 p.G310S). Patch-clamp recordings in transiently-transfected CHO cells and primary neurons revealed that both variants fully impeded Kv7.2 channel function, and exerted strong dominant-negative effects when co-expressed with Kv7.2 and/or Kv7.3 subunits. Notably, Kv7.2 p.G310S, but not Kv7.2 p.G310Δ10, currents were recovered upon overexpression of the PIP2-synthesizing enzyme PIP5K, and/or CaM; moreover, currents from heteromeric Kv7.2/Kv7.3 channels incorporating either Kv7.2 mutant subunits were differentially regulated by changes in PIP2 availability, with Kv7.2/Kv7.2 G310S/Kv7.3 currents showing a greater sensitivity to PIP2 depletion when compared to those from Kv7.2/Kv7.2 G310Δ10/Kv7.3 channels. Altogether, these results suggest that the two variants investigated differentially affected the splicing process at the intron 6-exon 7 boundary, and led to the synthesis of Kv7.2 subunits showing a differential sensitivity to PIP2 and CaM regulation; more studies are needed to clarify how such different functional properties contribute to the widely-divergent clinical phenotypes

The Central Andes and sustainable development: The Mendoza High Mountains Strategic Plan, Argentina

The Mendoza High Mountains Strategic Plan (MHMSP) is a key instrument for the public management and land-use planning of the Aconcagua region, Argentina. It involves stakeholders from government, the private sector, scientists and local communities in the Central Andes to promote collective action for making Aconcagua a sustainable tourism destination, within the context of climate change and COVID-19. The MHMSP is supported by the Federal Investment Council of Argentina and the Inter-American Development Bank. The Commission for Sustainable Development of the High Mountains of Mendoza comprises actors from the public and private sectors and was created to monitor the plan’s follow-up. Climate change adaptation and mitigation are part of the process outlined in the MHMSP, and support for the adoption and incorporation of new technologies plays a central role. Local communities whose livelihoods depend on snow tourism are vulnerable to the impact of climate change. Different mechanisms have been incorporated into the plan to promote mountain villages’ adaptation to climate change, to reduce their vulnerability.Fil: Reynoso, Marcelo. Gobierno de la Provincia de Mendoza. Ministerio de Turismo; ArgentinaFil: Rubio, María Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Salinas, Pamela. Provincia de Mendoza. Subsecretaría de Agricultura; ArgentinaFil: Morabito, Federico. Provincia de Mendoza. Subsecretaría de Agricultura; ArgentinaFil: Abraham, Elena Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Rubio, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Díaz, Fabián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Gil Fain Binda, Lorena. Provincia de Mendoza. Subsecretaría de Agricultura; ArgentinaFil: Zabala, Cecilia. Provincia de Mendoza. Subsecretaría de Agricultura; ArgentinaFil: Lo Bello, Silvia. Provincia de Mendoza. Subsecretaría de Agricultura; ArgentinaFil: Betancourt, Pablo. Betancourt Photographic Studio; ArgentinaFil: García, Gustavo. Betancourt Photographic Studio; Argentin

Terapias Farmacológicas e Não Farmacológicas no Manejo da Dermatite Atópica em Crianças: Uma Revisão Abrangente

  In the management of atopic dermatitis in children, both pharmacological and non-pharmacological therapies play fundamental roles in addressing this skin condition. These therapies offer a variety of options to control symptoms and improve the quality of life of affected children. However, it is important to understand and carefully evaluate the efficacy, safety, and impact of each therapy, taking into consideration the individual needs of each child. Exploring pharmacological and non-pharmacological therapies in the management of atopic dermatitis in children requires a comprehensive review of the different available approaches. This includes considering the effectiveness of topical medications such as corticosteroids and immunomodulators in controlling inflammation and reducing symptoms of atopic dermatitis. Additionally, non-pharmacological therapies such as emollients, therapeutic baths, and stress management techniques can also play an important role in symptom relief and maintaining skin health. In addition to clinical efficacy, it is crucial to consider factors such as potential side effects of medications, tolerability, treatment adherence, and individual preferences of children and their caregivers. Similarly, when assessing non-pharmacological therapies, it is important to examine practicality, acceptability, and impact on the quality of life of the child and their family. Customizing the management of atopic dermatitis is essential to ensure positive outcomes. This involves a thorough understanding of the clinical history and specific needs of each child, allowing for an adapted therapeutic approach. A child-centered approach, taking into account their interests, needs, and preferences, can enhance treatment effectiveness and satisfaction with outcomes. In summary, a comprehensive review of pharmacological and non-pharmacological therapies in the management of atopic dermatitis in children is essential for providing quality care. By considering efficacy, safety, individual preferences, and treatment customization, healthcare professionals can help children address the challenges of atopic dermatitis and improve their quality of life.No manejo da dermatite atópica em crianças, tanto as terapias farmacológicas quanto as não farmacológicas desempenham papéis fundamentais na abordagem dessa condição cutânea. Essas terapias oferecem uma variedade de opções para controlar os sintomas e melhorar a qualidade de vida das crianças afetadas. No entanto, é importante entender e avaliar cuidadosamente a eficácia, segurança e impacto de cada terapia, levando em consideração as necessidades individuais de cada criança. Explorar as terapias farmacológicas e não farmacológicas no manejo da dermatite atópica em crianças requer uma revisão abrangente das diferentes abordagens disponíveis. Isso inclui considerar a eficácia dos medicamentos tópicos, como corticosteroides e imunomoduladores, em controlar a inflamação e reduzir os sintomas da dermatite atópica. Além disso, terapias não farmacológicas, como emolientes, banhos terapêuticos e técnicas de manejo do estresse, também podem desempenhar um papel importante no alívio dos sintomas e na manutenção da saúde da pele. Além da eficácia clínica, é crucial considerar fatores como possíveis efeitos colaterais dos medicamentos, tolerabilidade, adesão ao tratamento e preferências individuais das crianças e de seus cuidadores. Da mesma forma, ao avaliar as terapias não farmacológicas, é importante examinar a praticidade, aceitabilidade e impacto na qualidade de vida da criança e de sua família.   Personalizar o manejo da dermatite atópica é essencial para garantir resultados positivos. Isso envolve uma compreensão aprofundada da história clínica e das necessidades específicas de cada criança, permitindo uma abordagem terapêutica adaptada. Uma abordagem centrada na criança, que leve em consideração seus interesses, necessidades e preferências, pode aumentar a eficácia do tratamento e a satisfação com os resultados. Em resumo, uma revisão abrangente das terapias farmacológicas e não farmacológicas no manejo da dermatite atópica em crianças é essencial para fornecer cuidados de qualidade. Ao considerar a eficácia, segurança, preferências individuais e personalização do tratamento, os profissionais de saúde podem ajudar as crianças a enfrentar os desafios da dermatite atópica e melhorar sua qualidade de vida. &nbsp

Involvement of cancer stem cells in glioblastoma angiogenesis

It is widely accepted that glioblastoma (GBM) develops from cancer stem cells (CSCs), a subset of stem-like cells displaying high resistance to treatment. In fact, despite aggressive therapy, 90% of patients relapse within 2 cm from tumor edge. Our recent findings showed the existence of a CSC type, residing in GBM peritumor tissue (PCSCs), that bears distinct characteristics from CSCs of the tumor mass (GCSCs). It should be considered the possibility that, after surgical resection, PCSCs might represent a reservoir of cells able to recapitulate the tumor. In this setting, characterization of PCSCs appears to be crucial in order to identify novel effective therapeutic targets. Thus, our aim was to investigate GCSCs and PCSCs role in angiogenesis, a key event in both GBM and peritumor tissue, whose vasculature shows features similar to those found in the tumor mass. In particular, we analyzed, by immunocytochemistry (ICC), Western blotting or real-time PCR, the expression of molecules involved in hypoxia and angiogenesis, such as HIF1α, HIF2α, and VEGF along with its receptors (VEGFR1, VEGFR2). ICC has highlighted the presence and the specific localization of these molecules in both GCSCs and PCSCs. The two cell populations showed comparable levels of VEGF. The transcript of VEGFR1 was in general expressed at higher levels in GCSCs than in PCSCs, while VEGFR2 mRNA and protein did not show a unique trend of expression. The expression of VEGF and its receptors in both GCSCs and PCSCs suggests that, besides well-known paracrine loops, autocrine signalings are also involved in tumor angiogenesis. Moreover, the expression of angiogenesis markers in PCSCs suggests these cells to have a direct role in peritumor tissue new vessel formation. In this regard, PCSCs should be considered a promising therapeutic target to counteract the angiogenesis-supported tumor progression