SORL1 mutation in a Greek family with Parkinson's disease and dementia

Whole exome sequencing and linkage analysis were performed in a three generational pedigree of Greek origin with a broad phenotypic spectrum spanning from Parkinson’s disease and Parkinson’s disease dementia to dementia of mixed type (Alzheimer disease and vascular dementia). We identified a novel heterozygous c.G1135T (p.G379W) variant in SORL1 which segregated with the disease in the family. Mutation screening in sporadic Greek PD cases identified one additional individual with the mutation, sharing the same 12.8Mb haplotype. Our findings provide support for SORL1 mutations resulting in a broad range of additional phenotypes and warrants further studies in neurodegenerative diseases beyond AD

Relevance of biomarkers across different neurodegenerative

Background: The panel of fluid- and imaging-based biomarkers available for neurodegenerative disease research is growing and has the potential to close important gaps in research and the clinic. With this growth and increasing use, appropriate implementation and interpretation are paramount. Various biomarkers feature nuanced differences in strengths, limitations, and biases that must be considered when investigating disease etiology and clinical utility. For example, neuropathological investigations of Alzheimer’s disease pathogenesis can fall in disagreement with conclusions reached by biomarker-based investigations. Considering the varied strengths, limitations, and biases of different research methodologies and approaches may help harmonize disciplines within the neurodegenerative disease field. Purpose of review: Along with separate review articles covering fluid and imaging biomarkers in this issue of Alzheimer’s Research and Therapy, we present the result of a discussion from the 2019 Biomarkers in Neurodegenerative Diseases course at the University College London. Here, we discuss themes of biomarker use in neurodegenerative disease research, commenting on appropriate use, interpretation, and considerations for implementation across different neurodegenerative diseases. We also draw attention to areas where biomarker use can be combined with other disciplines to understand issues of pathophysiology and etiology underlying dementia. Lastly, we highlight novel modalities that have been proposed in the landscape of neurodegenerative disease research and care

The Membrane Fusion Step of Vaccinia Virus Entry Is Cooperatively Mediated by Multiple Viral Proteins and Host Cell Components

For many viruses, one or two proteins allow cell attachment and entry, which occurs through the plasma membrane or following endocytosis at low pH. In contrast, vaccinia virus (VACV) enters cells by both neutral and low pH routes; four proteins mediate cell attachment and twelve that are associated in a membrane complex and conserved in all poxviruses are dedicated to entry. The aim of the present study was to determine the roles of cellular and viral proteins in initial stages of entry, specifically fusion of the membranes of the mature virion and cell. For analysis of the role of cellular components, we used well characterized inhibitors and measured binding of a recombinant VACV virion containing Gaussia luciferase fused to a core protein; viral and cellular membrane lipid mixing with a self-quenching fluorescent probe in the virion membrane; and core entry with a recombinant VACV expressing firefly luciferase and electron microscopy. We determined that inhibitors of tyrosine protein kinases, dynamin GTPase and actin dynamics had little effect on binding of virions to cells but impaired membrane fusion, whereas partial cholesterol depletion and inhibitors of endosomal acidification and membrane blebbing had a severe effect at the later stage of core entry. To determine the role of viral proteins, virions lacking individual membrane components were purified from cells infected with members of a panel of ten conditional-lethal inducible mutants. Each of the entry protein-deficient virions had severely reduced infectivity and except for A28, L1 and L5 greatly impaired membrane fusion. In addition, a potent neutralizing L1 monoclonal antibody blocked entry at a post-membrane lipid-mixing step. Taken together, these results suggested a 2-step entry model and implicated an unprecedented number of viral proteins and cellular components involved in signaling and actin rearrangement for initiation of virus-cell membrane fusion during poxvirus entry

Multiple Phosphatidylinositol 3-Kinases Regulate Vaccinia Virus Morphogenesis

Poxvirus morphogenesis is a complex process that involves the successive wrapping of the virus in host cell membranes. We screened by plaque assay a focused library of kinase inhibitors for those that caused a reduction in viral growth and identified several compounds that selectively inhibit phosphatidylinositol 3-kinase (PI3K). Previous studies demonstrated that PI3Ks mediate poxviral entry. Using growth curves and electron microscopy in conjunction with inhibitors, we show that that PI3Ks additionally regulate morphogenesis at two distinct steps: immature to mature virion (IMV) transition, and IMV envelopment to form intracellular enveloped virions (IEV). Cells derived from animals lacking the p85 regulatory subunit of Type I PI3Ks (p85α−/−β−/−) presented phenotypes similar to those observed with PI3K inhibitors. In addition, VV appear to redundantly use PI3Ks, as PI3K inhibitors further reduce plaque size and number in p85α−/−β−/− cells. Together, these data provide evidence for a novel regulatory mechanism for virion morphogenesis involving phosphatidylinositol dynamics and may represent a new therapeutic target to contain poxviruses

Качество жизни больных нестабильной бронхиальной астмой: влияние комбинированной терапии

The aim of this study was to evaluate influence of combined therapy with Seretid-Multidisk on quality of life (QoL) in patients with brittle and moderate asthma (BA). The study involved 33 patients: 21 patients had moderate BA (the 1st group) and 12 ones had brittle BA (the 2nd group) who received 1500 meg of beclomethazone dipropionate daily or another inhaled steriod in the equal dose 2.5 months at least and kept the daily bronchial lability more than 40% for at least 50% of the latter 75-day period. The basic therapy included Seretid-Multidisk 50/250: 1 inhaled dose twice a day in the 1st group and 2 inhaled doses twice a day in the 2nd group.The combined therapy with Seretid-Multidisk abolished a difference between QoL in moderate and brittle asthmatic patients. Seretid-Multidisk effected to the psycho-emotional and social fields in the brittle BA patients more than to those in the moderate BA patients. The combined therapy improved the QoL in BA patients in 3 days on the treatment keeping the effect up to 3 months.Целью настоящей работы явилось изучение влияния комбинированной терапии серетидом мультидиском на качество жизни (КЖ) больных нестабильной и среднетяжелой бронхиальной астмой (БА).В исследовании приняли участие 33 пациента: 21 человек (1-я группа) — больные среднетяжелой БА, 12 человек (2-я группа) больные с нестабильной БА, которые не менее 2,5 мес получали терапию беклометазоном дипропионатом в дозе 1500 мкг/сут или любым другим иГКС в эквивалентной дозе и при этом сохраняли среднесуточную лабильность бронхов более 40% на протяжении не менее 50% времени за последние 75 дней. В качестве базисной терапии использовали серетид мультидиск 50/250: в 1-й группе по 1 ингаляции 2 раза в день, во 2-й — по 2 ингаляции 2 раза в день.На фоне комбинированной терапии серетидом мультидиском разница между КЖ больных нестабильной и среднетяжелой БА нивелируется. При нестабильной БА влияние серетида мультидиска на психоэмоциональную и социальную сферы больного более выражено, чем при среднетяжелом заболевании. Комбинированная терапия приводит к достоверному повышению КЖ больных БА уже на 3- й день лечения с сохранением положительной динамики в течение последующих 3 мес

Decreased expression of the Augmenter of Liver Regeneration results in increased apoptosis and oxidative damage in human-derived glioma cells

The mammalian growth factor erv1-like (GFER) gene encodes a sulfhydryl oxidase enzyme, named Augmenter of Liver Regeneration (ALR). Recently it has been demonstrated that ALR supports cell proliferation acting as an anti-apoptotic factor. This effect is determined by ALR ability to support the anti-apoptotic gene expression and to preserve cellular normoxic conditions. We recently demonstrated that the addition of recombinant ALR (rALR) in the culture medium of H2O2-treated neuroblastoma cells reduces the lethal effects induced by the hydrogen peroxide. Similar data have been reported in the regenerating liver tissue from partially hepatectomized rats treated with rALR. The purpose of the present study was to evaluate the effect of the GFER inhibition, via the degradation of the complementary mRNA by the specific siRNA, on the behaviour of the apoptosis (apoptotic gene and caspase expression and apoptotic cell number) and of the oxidative stress-induced parameters (reactive oxygen species (ROS), clusterin expression and mitochondrial integrity) in T98G glioma cells. The results revealed a reduction of (i) ALR, (ii) clusterin and (iii) bcl-2 and an increase of (iv) caspase-9, activated caspase-3, ROS, apoptotic cell number and mitochondrial degeneration. These data confirm the anti-apoptotic role of ALR and its anti-oxidative properties, and shed some light on the molecular pathways through which ALR modulates its biological effects

Сниженная экспрессия генов нейрогенеза как биомаркер болезни Паркинсона у носителей мутаций в гене GBA: валидация анализа данных транскриптомного исследования

The objective of the study was to validate our previous results obtained during the transcriptome analysis of the primary culture of peripheral blood macrophages in patients with Parkinson's disease associated with mutations in the GBA gene (GBA-PD) in that reduced expression of the neurogenesis genes EGR1 (early growth response protein 1), NR4A2 (nuclear receptor 4A2), JUNB (transcription factor jun-B) in patients with GBA-PD.Methods and materials. The study included 14 patients with GBA-PD, 15 GBA-carriers, 30 patients with Parkinson's disease (PD) and 44 persons of the control group. The assessment of relative mRNA level of neurogenesis genes EGR1, NR4A2, JUNB in peripheral blood mononuclear cells were carried out by real-time quantitative polymerase chain reaction (PCR) using TaqMan fluorescent probes or EvaGreen fluorescent DNA dye.Results. Relative mRNA level of the JUNB gene in peripheral blood mononuclears was decreased in the group of patients with GBA-PD compared to controls (p=0.034). We found out that the relative mRNA level of the NR4A2 gene in peripheral blood mononuclears was increased in the group of patients with GBA-carriers compared to GBA-PD, patients with PD and controls (p=0.0029, p=0.00045, p=0.0024 respectively). There were no statistically significant differences in the mRNA level of the EGR1 gene between all the study groups (p>0.05).Conclusion. GBA-PD is characterized by reduced expression of the JUNB gene compared to control and of the NR4A2 gene compared to GBA-carriers.Цель исследования — проведение валидации результатов, полученных нами ранее в ходе анализа транскриптома первичной культуры макрофагов периферической крови пациентов с болезнью Паркинсона, ассоциированной с мутациями в гене лизосомного фермента глюкоцереброзидазы GBA (GBA-БП), в котором была выявлена сниженная экспрессия генов нейрогенеза EGR1 (early growth response protein 1), NR4A2 (nuclear receptor 4A2), JUNB (transcription factor jun-B) у пациентов с GBA-БП.Методы и материалы. В исследование включены 14 пациентов с GBA-БП, 15 GBA-носителей, 30 пациентов с болезнью Паркинсона (БП) и 44 индивидуума контрольной группы. Оценка относительного уровня мРНК генов нейрогенеза EGR1, NR4A2, JUNB в мононуклеарах периферической крови проводилась методом количественной полимеразной цепной реакции (ПЦР) в режиме реального времени с использованием флюоресцентных зондов TaqMan или флуоресцентного ДНК-красителя EvaGreen.Результаты. Относительный уровень мРНК гена JUNB в мононуклеарах периферической крови был понижен в группе пациентов с GBA-БП по сравнению с контрольной группой (p = 0,034). Также было выявлено, что относительный уровень мРНК гена NR4A2 в мононуклеарах периферической крови был повышен среди GBA-носителей, по сравнению с пациентами с GBA-БП, пациентами с БП и контролем (p = 0,0029, p = 0,00045, p = 0,0024 соответственно). Статистически значимых различий в уровне мРНК гена EGR1 между всеми исследуемых группами выявлено не было (p>0,05).Заключение. GBA-БП характеризуется пониженной экспрессией гена JUNB по сравнению с контролем и гена NR4A2 относительно GBA-носителей

Structural Analysis of Papain-Like NlpC/P60 Superfamily Enzymes with a Circularly Permuted Topology Reveals Potential Lipid Binding Sites

NlpC/P60 superfamily papain-like enzymes play important roles in all kingdoms of life. Two members of this superfamily, LRAT-like and YaeF/YiiX-like families, were predicted to contain a catalytic domain that is circularly permuted such that the catalytic cysteine is located near the C-terminus, instead of at the N-terminus. These permuted enzymes are widespread in virus, pathogenic bacteria, and eukaryotes. We determined the crystal structure of a member of the YaeF/YiiX-like family from Bacillus cereus in complex with lysine. The structure, which adopts a ligand-induced, “closed” conformation, confirms the circular permutation of catalytic residues. A comparative analysis of other related protein structures within the NlpC/P60 superfamily is presented. Permutated NlpC/P60 enzymes contain a similar conserved core and arrangement of catalytic residues, including a Cys/His-containing triad and an additional conserved tyrosine. More surprisingly, permuted enzymes have a hydrophobic S1 binding pocket that is distinct from previously characterized enzymes in the family, indicative of novel substrate specificity. Further analysis of a structural homolog, YiiX (PDB 2if6) identified a fatty acid in the conserved hydrophobic pocket, thus providing additional insights into possible function of these novel enzymes