Micro-Vesicles of Moringa oleifera Seeds in Heterozygous Rats for DAT Gene: Effects of Oral Intake on Behavioral Profile and Hematological Parameters

Previous studies have shown multiple biological properties of Moringa oleifera, a plant native to Africa and Asia. In the present study, potential physiological properties of microvesicles extracted from Moringa oleifera seeds were assessed. For this purpose, we investigated behavioral profile and hematological parameters in a recent rat model characterized by dysregulation in dopamine transporter, a key regulator of dopaminergic system. Experimental design consisted of male Wistar-DAT rats aged between two and four months: wild-type (WT) (n = 5) and heterozygous (DATHET) (n = 4) control groups, which drank tap water; WT (n = 5) and DATHET (n = 6) groups which drank a solution of Moringa microvesicles and water (2: 68 mL per day), which was orally administered for two months. Rats were monitored for spontaneous locomotor activity on a 24/7 basis. In the early lit hours, treated DATHET subjects showed higher locomotor activity, proposing a sleep-delay effect of Moringa. In forced swimming test, WT subjects who took Moringa exhibited more depressive behavior. In DATHET rats, Moringa seemed to potentiate the struggle to find a way out, counteracting an initial panic. Hemoglobin and hematocrit underwent opposite changes in either genotype, supporting the opposite effects on behavioral phenotype observed. Future work is clearly needed to further explore these preliminary profiles

MEOX2 Regulates the Growth and Survival of Glioblastoma Stem Cells by Modulating Genes of the Glycolytic Pathway and Response to Hypoxia

The most widely accepted hypothesis for the development of glioblastoma suggests that glioblastoma stem-like cells (GSCs) are crucially involved in tumor initiation and recurrence as well as in the occurrence of chemo- and radio-resistance. Mesenchyme homeobox 2 (MEOX2) is a transcription factor overexpressed in glioblastoma, whose expression is negatively correlated with patient survival. Starting from our observation that MEOX2 expression is strongly enhanced in six GSC lines, we performed shRNA-mediated knock-down experiments in two different GSC lines and found that MEOX2 depletion resulted in the inhibition of cell growth and sphere-forming ability and an increase in apoptotic cell death. By a deep transcriptome analysis, we identified a core group of genes modulated in response to MEOX2 knock-down. Among these genes, the repressed ones are largely enriched in genes involved in the hypoxic response and glycolytic pathway, two strictly related pathways that contribute to the resistance of high-grade gliomas to therapies. An in silico study of the regulatory regions of genes differentially expressed by MEOX2 knock-down revealed that they mainly consisted of GC-rich regions enriched for Sp1 and Klf4 binding motifs, two main regulators of metabolism in glioblastoma. Our results show, for the first time, the involvement of MEOX2 in the regulation of genes of GSC metabolism, which is essential for the survival and growth of these cells

Seroprevalence of Ebola virus infection in Bombali District, Sierra Leone

A serosurvey of anti-Ebola Zaire virus nucleoprotein IgG prevalence was carried out among Ebola virus disease survivors and their Community Contacts in Bombali District, Sierra Leone. Our data suggest that the specie of Ebola virus (Zaire) responsible of the 2013-2016 epidemic in West Africa may cause mild or asymptomatic infection in a proportion of cases, possibly due to an efficient immune response

Plant microRNAs from Moringa oleifera Regulate Immune Response and HIV Infection

Traditional medicine is often chosen due to its affordability, its familiarity with patient's cultural practices, and its wider access to the local community. Plants play an important role in providing indispensable nutrients, while specific small RNAs can regulate human gene expression in a cross-kingdom manner. The aim of the study was to evaluate the effects of plant-enriched purified extract microRNAs from Moringa oleifera seeds (MO) on the immune response and on HIV infection. Bioinformatic analysis shows that plant microRNAs (p-miRs) from MO belonging to 18 conserved families, including p-miR160h, p-miR166, p-miR482b, p-miR159c, p-miR395d, p-miR2118a, p-miR393a, p-miR167f-3p, and p-miR858b are predicted to target with high affinity BCL2, IL2RA, TNF, and VAV1, all these being involved in the cell cycle, apoptosis, immune response and also in the regulation of HIV pathogenesis. The effects of MO p-miRs transfected into HIV+ PBMCs were analyzed and revealed a decrease in viability associated with an increase of apoptosis; an increase of T helper cells expressing Fas and a decrease of intracellular Bcl2 protein expression. Meanwhile no effects were detected in PBMCs from healthy donors. In CD4(+) T cells, transfection significantly reduced cell activation and modified the T cell differentiation, thereby decreasing both central and effector memory cells while increasing terminal effector memory cells. Interestingly, the p-miRs transfection induces a reduction of intracellular HIV p24 protein and a reduction of viral DNA integration. Finally, we evaluated the effect of synthetic (mimic) p-miR858b whose sequence is present in the MO p-miR pool and predicted to target VAV1, a protein involved in HIV-Nef binding. This protein plays a pivotal role in T cell antigen receptor (TCR) signaling, so triggering the activation of various pathways. The transfection of HIV+ PBMCs with the synthetic p-miR858b showed a reduced expression of VAV1 and HIV p24 proteins. Overall, our evidence defines putative mechanisms underlying a supplementary benefit of traditional medicine, alongside current antiretroviral therapy, in managing HIV infection in resource-limited settings where MO remains widely available

Platinum-(IV)-derivative satraplatin induced G2/M cell cycle perturbation via p53-p21(waf1/cip1)-independent pathway in human colorectal cancer cells

Platinum-(IV)-derivative satraplatin represents a new generation of orally available anti-cancer drugs that are under development for the treatment of several cancers. Understanding the mechanisms of cell cycle modulation and apoptosis is necessary to define the mode of action of satraplatin. In this study, we investigate the ability of satraplatin to induce cell cycle perturbation, clonogenicity loss and apoptosis in colorectal cancer (CRC) cells.Platinum-(IV)-derivative satraplatin represents a new generation of orally available anti-cancer drugs that are under development for the treatment of several cancers. Understanding the mechanisms of cell cycle modulation and apoptosis is necessary to define the mode of action of satraplatin. In this study, we investigate the ability of satraplatin to induce cell cycle perturbation, clonogenicity loss and apoptosis in colorectal cancer (CRC) cells

Expression profile of HERVs and inflammatory mediators detected in nasal mucosa as a predictive biomarker of COVID-19 severity

IntroductionOur research group and others demonstrated the implication of the human endogenous retroviruses (HERVs) in SARS-CoV-2 infection and their association with disease progression, suggesting HERVs as contributing factors in COVID-19 immunopathology. To identify early predictive biomarkers of the COVID-19 severity, we analyzed the expression of HERVs and inflammatory mediators in SARS-CoV-2-positive and -negative nasopharyngeal/oropharyngeal swabs with respect to biochemical parameters and clinical outcome.MethodsResiduals of swab samples (20 SARS-CoV-2-negative and 43 SARS-CoV-2-positive) were collected during the first wave of the pandemic and expression levels of HERVs and inflammatory mediators were analyzed by qRT-Real time PCR.ResultsThe results obtained show that infection with SARS-CoV-2 resulted in a general increase in the expression of HERVs and mediators of the immune response. In particular, SARS-CoV-2 infection is associated with increased expression of HERV-K and HERV-W, IL-1β, IL-6, IL-17, TNF-α, MCP-1, INF-γ, TLR-3, and TLR-7, while lower levels of IL-10, IFN-α, IFN-β, and TLR-4 were found in individuals who underwent hospitalization. Moreover, higher expression of HERV-W, IL-1β, IL-6, IFN-α, and IFN-β reflected the respiratory outcome of patients during hospitalization. Interestingly, a machine learning model was able to classify hospitalized vs not hospitalized patients with good accuracy based on the expression levels of HERV-K, HERV-W, IL-6, TNF-a, TLR-3, TLR-7, and the N gene of SARS-CoV-2. These latest biomarkers also correlated with parameters of coagulation and inflammation.DiscussionOverall, the present results suggest HERVs as contributing elements in COVID-19 and early genomic biomarkers to predict COVID-19 severity and disease outcome

Moderate-intensity statin therapy seems ineffective in primary cardiovascular prevention in patients with type 2 diabetes complicated by nephropathy:a multicenter prospective 8 years follow up study

Background: Although numerous studies and metanalysis have shown the beneficial effect of statin therapy in CVD secondary prevention, there is still controversy such the use of statins for primary CVD prevention in patients with DM. The purpose of this study was to evaluate the occurrence of total major adverse cardio-vascular events (MACE) in a cohort of patients with type 2 diabetes complicated by nephropathy treated with statins, in order to verify real life effect of statin on CVD primary prevention. Methods: We conducted an observational prospective multicenter study on 564 patients with type 2 diabetic nephropathy free of cardiovascular disease attending 21 national outpatient diabetes clinics and followed them up for 8 years. 169 of them were treated with statins (group A) while 395 were not on statins (group B). Results: Notably, none of the patients was treated with a high-intensity statin therapy according to last ADA position statement. Total MACE occurred in 32 patients from group A and in 68 patients from group B. Fatal MACE occurred in 13 patients from group A and in 30 from group B; nonfatal MACE occurred in 19 patients from group A and in 38 patients from group B. The analysis of the Kaplan-Meier survival curves showed a not statistically significant difference in the incidence of total (p 0.758), fatal (p 0.474) and nonfatal (p 0.812) MACE between the two groups. HbA1c only showed a significant difference in the incidence of MACE between the two groups (HR 1.201, CI 1.041-1.387, p 0.012). Conclusions: These findings suggest that, in a real clinical setting, moderate-intensity statin treatment is ineffective in cardiovascular primary prevention for patients with diabetic nephropathy

Lopinavir/Ritonavir and Darunavir/Cobicistat in Hospitalized COVID-19 Patients: Findings From the Multicenter Italian CORIST Study

Background: Protease inhibitors have been considered as possible therapeutic agents for COVID-19 patients. Objectives: To describe the association between lopinavir/ritonavir (LPV/r) or darunavir/cobicistat (DRV/c) use and in-hospital mortality in COVID-19 patients. Study Design: Multicenter observational study of COVID-19 patients admitted in 33 Italian hospitals. Medications, preexisting conditions, clinical measures, and outcomes were extracted from medical records. Patients were retrospectively divided in three groups, according to use of LPV/r, DRV/c or none of them. Primary outcome in a time-to event analysis was death. We used Cox proportional-hazards models with inverse probability of treatment weighting by multinomial propensity scores. Results: Out of 3,451 patients, 33.3% LPV/r and 13.9% received DRV/c. Patients receiving LPV/r or DRV/c were more likely younger, men, had higher C-reactive protein levels while less likely had hypertension, cardiovascular, pulmonary or kidney disease. After adjustment for propensity scores, LPV/r use was not associated with mortality (HR = 0.94, 95% CI 0.78 to 1.13), whereas treatment with DRV/c was associated with a higher death risk (HR = 1.89, 1.53 to 2.34, E-value = 2.43). This increased risk was more marked in women, in elderly, in patients with higher severity of COVID-19 and in patients receiving other COVID-19 drugs. Conclusions: In a large cohort of Italian patients hospitalized for COVID-19 in a real-life setting, the use of LPV/r treatment did not change death rate, while DRV/c was associated with increased mortality. Within the limits of an observational study, these data do not support the use of LPV/r or DRV/c in COVID-19 patients

Transcriptional regulation by thymosin alpha-1 in peripheral blood monuclear cell from HIV positive patients

La Thymosin alpha-1 (Tα1) mostra una varietà di effetti sulle cellule e sulle vie di segnale del sistema immunitario. La Tα1 ha attività immunomodulante sia nelle cellule T che nella maturazione delle cellule NK. Inoltre ha attività come effettore sulla maturazione dei linfociti attraverso la produzione di citochine e attività citotossica. Diversi studi clinici su pazienti con infezioni o altre malattie, hanno inoltre dimostrato l’alta tollerabilità e l’assoluta sicurezza della Tα1, senza effetti collaterali a tutte le dosi utilizzate. In aggiunta, la Tα1 ha la capacità di attivare le cellule dentritiche infette attraverso la via di segnale indotta dai recettori Toll, influenzando in questo modo l’equlibirio infiammatorio e aumentando l’espressione di antigeni virali, tumorali e il complesso maggiore di istocompatibilità (MHC)I. Tuttavia le informazioni riguardanti l’uso della Tα1 in pazienti con infezione da HIV è molto limitata. Lo scopo principale degli esperimenti eseguiti per questa tesi è stato quello di analizzare quali geni coinvolti della risposta immunitaria possono essere regolati dalla Tα1 in vitro in PBMCs ottenuti da pazienti HIV. Questo per ottenere nuove informazioni sugli effetti della Tα1 sull’attività trascrizionale in questi pazienti, per meglio capire la risposta immunitaria compromessa in individui HIV+ e un possibile uso della Tα1 in combinazione con l’ART per il controllo dell’infezione da HIV. Pazienti HIV+ sono stati arruolati in uno studio trasversale ed in vitro i loro PBMCs e quelli di donatori sani sono stati isolati. A fresco le cellule sono state utilizzate per analizzare in citometria a flusso l’espressione delle molecole di superficie CD4+ CD8+, le cellule sono state incubate con Tα1 (50μg/ml) per 48h per analizzare l’espressione genica utilizzando un “Human Autoimmune & Inflammatory response” microarray. Dai risultati ottenuti dall’analisi del microarray, l’espressione di alcuni geni modulati è stata quantificata in Real Time PCR. I risultati confermano la capacità della timosina di regolare, nei linfociti, la risposta trascrizionale di un elevato numero di geni coinvolti nella risposta immunitaria. Risultato interessante, la risposta trascrizionale indotta dal trattamento con la Tα1, è elevata sia nei pazienti HIV+ che nei donatori sani, ma il grado di modulazione è differente tra i due gruppi suggerendo un effetto differente dovuto alla condizione basale di attivazione cellulare dei PBMCs dei pazienti e dei donatori e la distribuzione delle cellule bersaglio della Tα1. La stimolazione della produzione di chemochine indotta dalla Tα1 in PBMCs di pazienti HIV+, indica un doppio ruolo di questo peptide nell’attività antivirale ed immunomodulatoria. Il fenotipo dei pazienti infetti mostra una percentuale elevata di cellule CD8+. Quindi, la diversa modulazione delle chemochine nei pazienti e nei donatori potrebbe essere dovuta ad un preferenziale bersaglio cellulare della Tα1. L’applicazione della Tα1, che è capace allo stesso tempo di agire come adiuvante immunitario e simultaneamente opporsi all’infezione, potrebbe rappresentare un interessante nuovo approccio all’infezione da HIV.Thymosin alpha-1 (Tα1) has shown a variety of effects on cells and pathways of the immune system. Tα1 has been shown to exert an immunomodulatory activity towards both T cell and NK maturation. Moreover, Tα1 has been shown to act on effector functions of mature lymphocytes, such as cytokine production and cytotoxic activity. Several clinical studies on patients with infectious or other diseases, have already demonstrated the high tolerability and the absolute safety of Tα1, without side effects at all used doses. In addition, Tα1 has the ability to activate infected dendritic cells through Toll-like receptor signalling, thus influencing the inflammation balance, and increasing the expression of tumour, viral, and major histocompatibility complex (MHC) I antigens. However, information regarding the use of Tα1 in patients with HIV infection is very limited. The principal aim of the experiments performed in the framework of this Ph.D. thesis was to investigate which genes belonging to immune response pathways could be regulated by Tα1 in vitro in PBMCs from HIV infected individuals. This, to obtain new information on the effect of Tα1 on the transcriptional activity in these patients and to further understand the impaired immune response in PBMCs from HIV+ patients and a possible helpfulness of the use Tα1 in combination with antiretroviral therapy for the control of HIV infection. HIV+ individuals were enrolled in an open cross-sectional study. In vitro PBMCs from HIV+ individuals and from healthy donors were isolated. Fresh cells were used for flow cytometric analysis of cellular proteins (CD4+ CD8+), cells were incubated with Tα1 (50μg/ml) for 48h to evaluate gene expression profile using Human Autoimmune & Inflammatory response microarray system. Based on the results of microarray analysis, the expression of genes modulated was quantified using Real Time PCR. The results confirm the ability of Tα1 to regulate, in lymphoid cells, the transcriptional response of a high number of genes involved in the immune response. Interestingly, the transcriptional response to Tα1 treatment is elevated in HIV+ patients as well as in healthy donors, but the degree of modulation was different between the two groups, suggesting a differential effect owing to the basal condition of cellular activation of the PBMCs from HIV+ patients and healthy donors and the distribution of Tα1 targeted cells. The stimulation of the production of chemokines by Tα1 in PBMCs from HIV+ patients indicates for a dual role of this peptide in the simultaneously immune modulator and antiviral action. The phenotype of HIV+ PBMCs showed an higher percentage of CD8+ cell. Thus, the different modulation of chemokines in the patients and healthy donors, might be due to the preferential cell target of Tα1. Taking in account the impossibility to eradicate the virus from the reservoir, the ART therapy is able at the moment to inhibit the virus replication and promote the recovery of T cell compartment. The application of Tα1 that could be able to act at the same time as immune adjuvant and simultaneously opposite the infection would represent an interesting additional approach to HIV infection

High CD169 Monocyte/Lymphocyte Ratio Reflects Immunophenotype Disruption and Oxygen Need in COVID-19 Patients

Background: Sialoadhesin (CD169) has been found to be overexpressed in the blood of COVID-19 patients and identified as a biomarker in early disease. We analyzed CD169 in the blood cells of COVID-19 patients to assess its role as a predictive marker of disease progression and clinical outcomes. Methods: The ratio of the median fluorescence intensity of CD169 between monocytes and lymphocytes (CD169 RMFI) was analyzed by flow cytometry in blood samples of COVID-19 patients (COV) and healthy donors (HDs) and correlated with immunophenotyping, inflammatory markers, cytokine mRNA expression, pulmonary involvement, and disease progression. Results: CD169 RMFI was high in COV but not in HDs, and it correlated with CD8 T-cell senescence and exhaustion markers, as well as with B-cell maturation and differentiation in COV. CD169 RMFI correlated with blood cytokine mRNA levels, inflammatory markers, and pneumonia severity in patients who were untreated at sampling, and was associated with the respiratory outcome throughout hospitalization. Finally, we also report the first evidence of the specific ability of the spike protein of SARS-CoV-2 to trigger CD169 RMFI in a dose-dependent manner in parallel with IL-6 and IL-10 gene transcription in HD PBMCs stimulated in vitro. Conclusion: CD169 is induced by the spike protein and should be considered as an early biomarker for evaluating immune dysfunction and respiratory outcomes in COVID-19 patients