179. Helper-Dependent Adenovirus-Mediated Gene Transfer of an LDL Receptor/Transferrin Chimeric Protein Reduces Aortic Atherosclerosis in LDL Receptor-Deficient Mice

Familial hypercholesterolemia (FH) is a well-characterized genetic hyperlipidemia due in most of the cases to mutations in the LDL receptor (LDLR) gene; FH is characterized by elevated concentration of plasma LDL cholesterol (LDL-C) with consequent deposition of LDL-C in tendons, skin and arteries. Statins can lower cholesterol levels but are not effective in all patients whose prognosis is still quite poor. In the past, we have developed safe and effective gene-therapy strategies for hepatocytes transduction and consequent expression of anti-atherogenic proteins using PEGylated helper-dependent adenoviral (HD-Ad) vectors. We have recently devised a therapeutic strategy for reducing LDL using a secreted protein that can potentially be expressed in non-hepatic tissues used as bioreactors. At this aim, we developed an HD-Ad vector for the expression of the soluble form of the extracellular portion of the human LDLR fused with transferrin (LDLR/Tf). We evaluated the efficacy of LDLR/Tf in cellular models such as CHOldla7 in which we restored the cell ability to uptake of labeled LDL; subsequently, we administered intravenously 1X10E11 vp/kg of the HD-Ad vector expressing LDLR/Tf in LDLR-deficient mice and demonstrated the efficacy of the above-mentioned vector in reducing total and LDL cholesterol levels; in addition, expression of LDLR/Tf significantly reduced aortic atherosclerotic lesions in treated LDLR-deficient mice compared to controls 1.78±0.48 vs. 5.38±0.54 sq.mm.). We therefore demonstrated the efficacy of serum secretion of LDLR/Tf in reducing aortic atherosclerosis in FH mice. These results will allow the evaluation of HD-Ad vector-mediated expression of LDLR/Tf in non-hepatic tissues using alternative routes of administration in order to develop safer gene transfer protocol more compatible with clinical applications

Типологія синтаксичних конструкцій в німецькій та українській мовах

Німецька та українська мови є односистемними мовами: обидві належать до індоєвропейської мовної сім’ї. Спільні корені та тривалий період ізольованого розвитку, вказують на те, що вказані мови мають характеристики подібності та відмінності в своій внутрішній будові. Німецька та українська належать до синтетичного типу флективних мов. Це означає, що граматичне значення слів у них виражається, здебільшого, за допомогою системи флексій і реалізується в межах одного графічного слова. Але флективна система німецької мови бідніша, ніж у слов’янських мовах.Немецкий и украинский языки являются односистемными языками: оба принадлежат к индоевропейской языковой семье. Общие корни и длительный период изолированного развития, указывают на то, что указанные языки имеют характеристики сходства и различия в своем внутреннем строении. Немецкий и украинский принадлежат к синтетическому типу флективных языков. Это означает, что грамматическое значение слов в них выражается, в основном, с помощью системы флексий и реализуется в пределах одного графического слова. Но флективная система немецкого языка беднее, чем в славянских языках.German and Ukrainian are single-system languages: both belong to the Indo-European language family. Common roots and a long period of isolated development, indicate that these languages ​​have characteristics of similarity and differences in their internal structure. German and Ukrainian belong to the synthetic type of inflectional languages. This means that the grammatical meaning of words in them is expressed, mainly, with the help of a system of inflexions and is realized within a single graphic word. But the inflectional system of the German language is poorer than in the Slavic languages

Celiac disease-associated Neisseria flavescens decreases mitochondrial respiration in CaCo-2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial-induced cellular imbalance

: We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo-2 cells. We also found that vesicular trafficking was delayed after the CD-immunogenic P31-43 gliadin peptide-entered CaCo-2 cells and that Lactobacillus paracasei CBA L74 (L. paracasei-CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD-N. flavescens-infected CaCo-2 cells and if any alteration could be mitigated by pretreating cells with L. paracasei-CBA supernatant, despite the presence of P31-43. We measured CaCo-2 bioenergetics by an extracellular flux analyser, N. flavescens and P31-43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD-N. flavescens colocalised more than control N. flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31-43 increased colocalisation of N. flavescens with early vesicles. Mitochondrial respiration was lower (P < .05) in CD-N. flavescens-infected cells versus not-treated CaCo-2 cells, whereas pretreatment with L. paracasei-CBA reduced CD-N. flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD-N. flavescens induces metabolic imbalance in CaCo-2 cells, and the L. paracasei-CBA probiotic could be used to correct CD-associated dysbiosis

Risk of Getting COVID-19 in People With Multiple Sclerosis

Background and Objectives Several studies have assessed risk factors associated with the severity of COVID-19 outcomes in people with multiple sclerosis (PwMS). The potential role of disease-modifying therapies (DMTs) and demographic and clinical factors on the risk of acquiring SARS-CoV-2 infection has not been evaluated so far. The objective of this study was to assess risk factors of contracting SARS-CoV-2 infection in PwMS by using data collected in the Italian MS Register (IMSR). Methods Acase-control (1:2) studywas set up. Cases included PwMSwith a confirmed diagnosis ofCOVID-19, and controls included PwMS without a confirmed diagnosis of COVID-19. Both groups were propensity score–matched by the date of COVID-19 diagnosis, the date of last visit, and the region of residence. No healthy controls were included in this study. COVID-19 risk was estimated by multivariable logistic regression models including demographic and clinical covariates. The impact of DMTs was assessed in 3 independent logistic regression models including one of the following covariates: last administeredDMT, previousDMTsequences, or the place where the last treatment was administered. Results A total of 779 PwMS with confirmed COVID-19 (cases) were matched to 1,558 PwMS without COVID-19 (controls). In all 3 models, comorbidities, female sex, and a younger age were significantly associated (p < 0.02)with a higher risk of contractingCOVID-19. Patients receiving natalizumab as last DMT(OR[95%CI]: 2.38 [1.66–3.42], p < 0.0001) and those who underwent an escalation treatment strategy (1.57 [1.16–2.13], p = 0.003) were at significantly higher COVID-19 risk. Moreover, PwMS receiving their last DMT requiring hospital access (1.65 [1.34–2.04], p < 0.0001) showed a significant higher risk than those taking self-administered DMTs at home. Discussion This case-control study embedded in the IMSR showed that PwMS at higher COVID-19 risk are younger, more frequently female individuals, and with comorbidities. Long-lasting escalation approach and last therapies that expose patients to the hospital environment seem to significantly increase the risk of SARS-CoV2 infection in PwMS. Classification of Evidence This study provides Class III evidence that among patients with MS, younger age, being female individuals, having more comorbidities, receiving natalizumab, undergoing an escalating treatment strategy, or receiving treatment at a hospital were associated with being infected with COVID-19. Among patients with MS who were infected with COVID-19, a severe course was associated with increasing age and having a progressive form of MS, whereas not being on treatment or receiving an interferon beta agent was protective

Plio-Pleistocene exhumation of the eastern Himalayan syntaxis and its domal ‘pop-up’

The eastern termination of the Himalayan orogen forms a structural syntaxis that is characterised by young (from 10 to < 1 Ma) mineral growth and cooling ages that document Late Miocene to Pleistocene structural, metamorphic, igneous and exhumation events. This region is a steep antiformal and in part domal structure that folds the suture zone between the Indian and Asian plates. It is dissected by the Yarlung Tsangpo, one of the major rivers of the eastern Himalayan–Tibet region, which becomes the Brahmaputra River in the Indian foreland basin before emptying into the Bay of Bengal. Exceptionally high relief and one of the deepest gorges on Earth have developed where the river's tortuous route crosses the Namche Barwa–Gyala Peri massif (> 7 km in elevation) in the core of the syntaxis. Very high erosion rates documented in sediment downstream of the gorge at the foot of the Himalaya contribute ~ 50% of total detritus to the sediment load of the Brahmaputra. The initiation of very high rates of exhumation has been attributed either to the extreme erosive power of a river flowing across a deforming indentor corner and the associated positive feedback, or to the geometry of the Indian plate indentor, with the corner being thrust beneath the Asian plate resulting in buckling which accommodates shortening; both processes may be important. The northern third of the syntaxis corresponds to a steep domal ‘pop-up’ structure bounded by the India–Asia suture on three sides and a thrust zone to the south. Within the dome, Greater Himalaya rocks equilibrated at ~ 800 °C and 25–30 km depth during the Miocene, with these conditions potentially persisting into the latest Miocene and possibly the Pliocene, with modest decompression prior to ~ 4 Ma. This domal ‘pop-up’ corresponds to the area of youngest bedrock ages on a wide variety of thermochronometers and geochronometers. In this paper we review the extensive scientific literature that has focused on the eastern syntaxis and provide new chronological data on its bedrock and erosion products to constrain the age of inception of the very rapid uplift and erosion. We then discuss its cause, with the ultimate aim to reconstruct the exhumation history of the syntaxis and discuss the tectonic context for its genesis. We use zircon and rutile U–Pb, white mica Ar–Ar and zircon fission track dating methods to extract age data from bedrock, Brahmaputra modern sediments (including an extensive compilation of modern detrital chronometry from the eastern Himalaya) and Neogene palaeo-Brahmaputra deposits of the Surma Basin (Bangladesh). Numerical modelling of heat flow and erosion is also used to model the path of rocks from peak metamorphic conditions of ~ 800 °C to < 250 °C. Our new data include U–Pb bedrock rutile ages as young as 1.4 Ma from the Namche Barwa massif and 0.4 Ma from the river downstream of the syntaxis. Combined with existing data, our new data and heat flow modelling show that: i) the detrital age signature of the modern syntaxis is unique within the eastern Himalayan region; ii) the rocks within the domal pop-up were > 575 ± 75 °C only 1–2 Myr ago; iii) the Neogene Surma Basin does not record evidence of the rise and erosion of the domal pop-up until latest Pliocene–Pleistocene time; iv) Pleistocene exhumation of the north-easternmost part of the syntaxis took place at rates of at least 4 km/Myr, with bedrock erosion of 12–21 km during the last 3 Ma; v) the inception of rapid syntaxial exhumation may have started as early as 7 Ma or as late as 3 Ma; and vi) the Yarlung Tsangpo is antecedent and subsequently distorted by the developing antiform. Together our data and modelling demonstrate that the domal pop-up with its exceptional erosion and topographic relief is likely a Pleistocene feature that overprinted earlier structural and metamorphic events typical of Himalayan evolution. Keywords: Eastern Himalayan syntaxis; Namche Barwa; Surma Basin; Yarlung Tsangpo–Brahmaputra; U–Pb rutile dating; Thermal modellin

Disease-Modifying Therapies and Coronavirus Disease 2019 Severity in Multiple Sclerosis

Objective: This study was undertaken to assess the impact of immunosuppressive and immunomodulatory therapies on the severity of coronavirus disease 2019 (COVID-19) in people with multiple sclerosis (PwMS). Methods: We retrospectively collected data of PwMS with suspected or confirmed COVID-19. All the patients had complete follow-up to death or recovery. Severe COVID-19 was defined by a 3-level variable: mild disease not requiring hospitalization versus pneumonia or hospitalization versus intensive care unit (ICU) admission or death. We evaluated baseline characteristics and MS therapies associated with severe COVID-19 by multivariate and propensity score (PS)-weighted ordinal logistic models. Sensitivity analyses were run to confirm the results. Results: Of 844 PwMS with suspected (n = 565) or confirmed (n = 279) COVID-19, 13 (1.54%) died; 11 of them were in a progressive MS phase, and 8 were without any therapy. Thirty-eight (4.5%) were admitted to an ICU; 99 (11.7%) had radiologically documented pneumonia; 96 (11.4%) were hospitalized. After adjusting for region, age, sex, progressive MS course, Expanded Disability Status Scale, disease duration, body mass index, comorbidities, and recent methylprednisolone use, therapy with an anti-CD20 agent (ocrelizumab or rituximab) was significantly associated (odds ratio [OR] = 2.37, 95% confidence interval [CI] = 1.18-4.74, p = 0.015) with increased risk of severe COVID-19. Recent use (<1 month) of methylprednisolone was also associated with a worse outcome (OR = 5.24, 95% CI = 2.20-12.53, p = 0.001). Results were confirmed by the PS-weighted analysis and by all the sensitivity analyses. Interpretation: This study showed an acceptable level of safety of therapies with a broad array of mechanisms of action. However, some specific elements of risk emerged. These will need to be considered while the COVID-19 pandemic persists

COVID-19 Severity in Multiple Sclerosis: Putting Data Into Context

Background and objectives: It is unclear how multiple sclerosis (MS) affects the severity of COVID-19. The aim of this study is to compare COVID-19-related outcomes collected in an Italian cohort of patients with MS with the outcomes expected in the age- and sex-matched Italian population. Methods: Hospitalization, intensive care unit (ICU) admission, and death after COVID-19 diagnosis of 1,362 patients with MS were compared with the age- and sex-matched Italian population in a retrospective observational case-cohort study with population-based control. The observed vs the expected events were compared in the whole MS cohort and in different subgroups (higher risk: Expanded Disability Status Scale [EDSS] score > 3 or at least 1 comorbidity, lower risk: EDSS score ≤ 3 and no comorbidities) by the χ2 test, and the risk excess was quantified by risk ratios (RRs). Results: The risk of severe events was about twice the risk in the age- and sex-matched Italian population: RR = 2.12 for hospitalization (p < 0.001), RR = 2.19 for ICU admission (p < 0.001), and RR = 2.43 for death (p < 0.001). The excess of risk was confined to the higher-risk group (n = 553). In lower-risk patients (n = 809), the rate of events was close to that of the Italian age- and sex-matched population (RR = 1.12 for hospitalization, RR = 1.52 for ICU admission, and RR = 1.19 for death). In the lower-risk group, an increased hospitalization risk was detected in patients on anti-CD20 (RR = 3.03, p = 0.005), whereas a decrease was detected in patients on interferon (0 observed vs 4 expected events, p = 0.04). Discussion: Overall, the MS cohort had a risk of severe events that is twice the risk than the age- and sex-matched Italian population. This excess of risk is mainly explained by the EDSS score and comorbidities, whereas a residual increase of hospitalization risk was observed in patients on anti-CD20 therapies and a decrease in people on interferon

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon

A specific immunophenotype and an increased adipogenic potential characterized human amniotic mesenchymal stem cells (hA-MSCs) isolated from obese pregnant women at delivery

Maternal obesity is associated to increased fetal risk of obesity and other metabolic diseases. Human amniotic mesenchymal stem cells (hA-MSC) have not been characterized in obese women. The aim of this study was to isolate and compare hA-MSC immunophenotypes from obese (Ob-) and normal weight control (Co-) women to identify alterations possibly predisposing the foetus to obesity. We enrolled 16 Ob- and 7 Co-women at delivery (mean/SEM pre-pregnancy BMI: 40.3/1.8 kg/m2 and 22.4/1.0 kg/m2, respectively). hA-MSCs were phenotyped by flow cytometry; several maternal and newborn clinical and biochemical parameters were also measured. The expression of membrane antigen CD13 was higher on Ob-hA-MSCs than on Co-hA-MSCs (P=0.0043). Also serum levels of CD13 at delivery were higher in Ob- versus Co-pregnant women and correlated with CD13 antigen expression on Ob-hA-MSCs (r2=0.84, P<0.0001). Adipogenesis induction experiments revealed that Ob-hA-MSCs had a higher adipogenic potential than Co-hA-MSCs as witnessed by higher PPARg and aP2 mRNA levels (P=0.02 and P=0.03, respectively) at post-induction day 14 associated with an increase of CD13 mRNA from baseline to day 4 post-induction (P<0.05). Adipogenesis was similar in the two sets of hA-MSCs after CD13 silencing, whereas it was increased in Co-hA-MSCs after CD13 overexpression. Pretreatment of Co- and Ob-hA-MSCs with IFN-g ng/mL) for 24 h induced upregulation of CD13 expression. In conclusion, antigen CD13, by influencing the adipogenic potential of hA-MSCs could be an in-utero risk factor for obesity. Our data strengthen the hypothesis that high levels of serum and MSC CD13 are obesity markers