Development of FRET-based high-throughput screening for viral RNase III inhibitors

The class 1 ribonuclease III (RNase III) encoded by Sweet potato chlorotic stunt virus (CSR3) suppresses RNA silencing in plant cells and thereby counters the host antiviral response by cleaving host small interfering RNAs, which are indispensable components of the plant RNA interference (RNAi) pathway. The synergy between sweet potato chlorotic stunt virus and sweet potato feathery mottle virus can reduce crop yields by 90%. Inhibitors of CSR3 might prove efficacious to counter this viral threat, yet no screen has been carried out to identify such inhibitors. Here, we report a novel high-throughput screening (HTS) assay based on fluorescence resonance energy transfer (FRET) for identifying inhibitors of CSR3. For monitoring CSR3 activity via HTS, we used a small interfering RNA substrate that was labelled with a FRET-compatible dye. The optimized HTS assay yielded 109 potential inhibitors of CSR3 out of 6,620 compounds tested from different small-molecule libraries. The three best inhibitor candidates were validated with a dose-response assay. In addition, a parallel screen of the selected candidates was carried out for a similar class 1 RNase III enzyme from Escherichia coli (EcR3), and this screen yielded a different set of inhibitors. Thus, our results show that the CSR3 and EcR3 enzymes were inhibited by distinct types of molecules, indicating that this HTS assay could be widely applied in drug discovery of class 1 RNase III enzymes.Peer reviewe

Changes in Institutionalized Older People's Dentition Status in Helsinki, 2003-2017

Non peer reviewe

Anti-neuronal anti-bodies in patients with early psychosis

It may be challenging to distinguish autoimmune encephalitis associated with anti-neuronal autoantibodies from primary psychiatric disorders. Here, serum was drawn from patients with a first-episode psychosis (n = 70) or a clinical high-risk for psychosis (n = 6) and controls (n = 34). We investigated the serum prevalence of 24 antineuronal autoantibodies: IgG antibodies for anti-N-methyl-D-aspartate-type glutamate receptor (anti-NMDAR), glutamate and gamma-aminobutyric acid alpha and beta receptors (GABA-a, GABA-b), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA), glycine receptor (GlyR), metabotropic glutamate receptor 1 and 5 (mGluR1, mGluR5), anti-Tr/Delta/notch-like epidermal growth factor-related receptor (DNER), contactin-associated protein-like 2 (CASPR2), myelin oligodendrocyte glycoprotein (MOG), glutamic acid decarboxylase-65 (GAD65), collapsin response mediator protein 5/crossveinless-2 (CV2), aquaporin-4 (AQP4), anti-dipeptidyl-peptidase-like protein-6 (DPPX), type 1 anti-neuronal nuclear antibody (ANNA-1, Hu), Ri, Yo, IgLON5, Ma2, zinc finger protein 4 (ZIC4), Rho GTPase-activating protein 26, amphiphysin, and recoverin, as well as IgA and IgM for dopamine-2-receptor (DRD2). Anti-NMDA IgG antibodies were positive with serum titer 1:320 in one patient with a clinical high risk for psychosis. He did not receive a diagnosis of encephalitis after comprehensive neurological evaluation. All other antineuronal autoantibodies were negative and there were no additional findings with immunohistochemistry of brain issues. (C) 2017 Elsevier B.V. All rights reserved.Peer reviewe

Oral disease burden of dentate older adults living in long-term care facilities: FINORAL study

Background A growing number of older adults have natural teeth and are at high risk of oral diseases, which are induced by oral bacterial accumulation and proceed unnoticed and quietly. Our aim was (1) to examine the association of oral disease burden (ODB) with health and functioning among dentate long-term care residents, and (2) to find easily detectable signs for nurses to identify residents' poor oral health. Methods In this cross-sectional observational study dentists examined 209 residents' oral status, and nurses assessed residents for their functioning and nutrition in long-term care facilities in Helsinki, Finland. ODB was defined by asymptotic dental score (ADS). Six clinical signs of residents' poor oral health were considered as potentially easy for nurses to detect: lesions on lips, teeth with increased mobility, lesions on oral mucosa, eating soft or pureed food, unclear speech, and needing assistance in eating. The association of these was tested with high ODB as outcome. Results Participants were grouped according to their ADS scores: low (n = 39), moderate (n = 96) and high ODB (n = 74). ODB was linearly associated with coronary artery disease and poor cognitive and physical functioning: needing assistance in eating, poor ability to make contact, and unclear speech but not with other diseases including dementia or demographic characteristics. Furthermore, ODB was linearly associated with eating soft or pureed food. Of the six selected, easily detectable signs, having at least two positive signs gave 89% sensitivity to detecting high ODB. Conclusion Poor oral health was common and ODB accumulated among residents with poor functioning. Nurses may use a few easily detectable signs to screen residents' oral health when considering a resident's need for consultation with an oral health professional.Peer reviewe

In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III

Sweet potato virus disease (SPVD), caused by synergistic infection of Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV), is responsible for substantial yield losses all over the world. However, there are currently no approved treatments for this severe disease. The crucial role played by RNase III of SPCSV (CSR3) as an RNA silencing suppressor during the viruses' synergistic interaction in sweetpotato makes it an ideal drug target for developing antiviral treatment. In this study, high-throughput screening (HTS) of small molecular libraries targeting CSR3 was initiated by a virtual screen using Glide docking, allowing the selection of 6,400 compounds out of 136,353. We subsequently developed and carried out kinetic-based HTS using fluorescence resonance energy transfer technology, which isolated 112 compounds. These compounds were validated with dose-response assays including kinetic-based HTS and binding affinity assays using surface plasmon resonance and microscale thermophoresis. Finally, the interference of the selected compounds with viral accumulation was verified in planta. In summary, we identified five compounds belonging to two structural classes that inhibited CSR3 activity and reduced viral accumulation in plants. These results provide the foundation for developing antiviral agents targeting CSR3 to provide new strategies for controlling sweetpotato virus diseases. IMPORTANCE We report here a high-throughput inhibitor identification method that targets a severe sweetpotato virus disease caused by coinfection with two viruses (SPCSV and SPFMV). The disease is responsible for up to 90% yield losses. Specifically, we targeted the RNase III enzyme encoded by SPCSV, which plays an important role in suppressing the RNA silencing defense system of sweetpotato plants. Based on virtual screening, laboratory assays, and confirmation in planta, we identified five compounds that could be used to develop antiviral drugs to combat the most severe sweetpotato virus disease.Peer reviewe

Relationship between Fried's frailty phenotype and oral frailty in long-term care residents

Background: oral frailty (OFr) may be called a syndrome lacking a consensus on its definition. Objective: the aim was to prove the relationship between OFr to the phenotype of frailty, general health and nutrition in long-term care. Design: the FINnish ORAL Health Study in Long-Term Care study is a cross-sectional clinical research comprising findings on oral and general health and nutrition. Setting: participants were divided into groups according to the number of OFr signs: Group 1 (0-1 sign), Group 2 (2-4 signs) and Group 3 (5-6 signs). Subjects: the study includes data on 349 older residents of long-term care facilities in Helsinki, Finland. Methods: frailty status was defined according to Fried's frailty phenotype. OFr was evaluated with six signs: dry mouth, diet of pureed or soft food, residue of food on oral surfaces, unclear speech, inability to keep mouth open during the clinical oral examination and pain expression during the examination. Results: a significant linear relationship across the OFr groups with Fried's frailty phenotype was found (P for linearity = 0.008, adjusted by gender and age). A linear trend existed between OFr groups and general health; prevalence of dementia and malnutrition increased from Group 1 to Group 3. The need for help with eating and oral hygiene procedures increased from Group 1 to Group 3. Moreover, OFr had a linear relationship with chewing and swallowing difficulties. Conclusions: OFr is related to Fried's frailty phenotype, general health, nutrition and need for help with daily activities.Peer reviewe

Oral hygiene and health-related quality of life in institutionalized older people

Purpose We evaluated the level of oral hygiene and its association with oral health status and need for oral treatment among older residents in long-term care facilities. In addition, the association between oral hygiene level and health-related quality of life (HRQoL) was explored. Methods This cross-sectional study assessed 231 dentate residents in long-term care facilities (71% female, mean age 81 years, 70% had dementia). Nurses assessed residents and completed questionnaires on participants' background information, diagnoses, oral healthcare habits, and HRQoL with the 15D instrument. Two qualified dentists performed clinical oral examinations (number of teeth, plaque index, periodontal condition, open caries lesions, and dry mouth). We used a modified plaque index (PI) to measure the level of oral hygiene (good, moderate, and poor) and calculated the clinical Asymptotic Dental Score (ADS) to determine the oral inflammation burden. Results Of the residents, 21% had good, 35% moderate, and 44% poor oral hygiene according to PI. Poor oral hygiene was associated with poorer cognitive status (P = 0.010) and higher oral inflammation burden (P < 0.001). Moreover, poor oral hygiene was associated with poorer HRQoL in a correlation analysis adjusted for age and gender. Conclusions Oral hygiene of older individuals in long-term care is insufficient. Poor oral hygiene is a marker for poor HRQoL. Residents also have a high burden of oral inflammatory diseases and a need for dental care. Older residents' oral hygiene and HRQoL may be improved with oral care education of caregivers and regular dental check-ups. Key summary pointsAim We evaluated oral hygiene level and its association with oral health and general health-related quality of life (HRQoL) among older residents in long-term care facilities. Findings Only one-fifth of residents had good oral hygiene. Poor oral hygiene was associated with poor oral health and diminished HRQoL. Message Oral hygiene, oral health, and HRQoL may be improved with oral care education of caregivers, professional cooperation, and regular oral healthcare of older residents in long-term care facilities.Peer reviewe

A design for neural network model of continuous reading

Cognition and learning are exceedingly modeled as an associative activity of connectionist neural networks. However, only a few such models exist for continuous reading, which involves the delicate coordination of word recognition and eye movements. Moreover, these models are limited to only orthographic level of word processing with predetermined lexicons. Here, we present a conceptual design of a developmentally plausible neural network model of reading designed to simulate word learning, parafoveal preview activation of words, their later foveal word recognition including phonological decoding, and forward saccade length as a control mechanism for intake of new textual information. We will discuss the theoretical advancements of the design and avenues for future developments

Concentration Dependence of the Effective Mass of He-3 Atoms in He-3/He-4 Mixtures

Recent measurements by Yorozu et al. (S. Yorozu, H. Fukuyama, and H. Ishimoto, Phys. Rev. B 48, 9660 (1993)) as well as by Simons and Mueller (R. Simons and R. M. Mueller, Czhechoslowak Journal of Physics Suppl. 46, 201 (1976)) have determined the effective mass of He-3 atoms in a He-3/He-4 mixture with great accuracy. We here report theoretical calculations for the dependence of that effective mass on the He-3 concentration. Using correlated basis functions perturbation theory to infinite order to compute effective interactions in the appropriate channels, we obtain good agreement between theory and experiment.Comment: 4 pages, 1 figur

Burden of Oral Symptoms and Health-Related Quality of Life in Long-Term Care Settings in Helsinki, Finland

Objectives Poor oral health may complicate eating and deteriorate nutritional status. However, little is known about how the burden of oral symptoms (OS) is associated with the health-related quality of life (HRQoL) of vulnerable older people in institutional settings. This study explores how the burden of certain OS (chewing problems, swallowing difficulties, dry mouth) is associated with functioning, morbidity, nutritional status and eating habits. It also examines the association between the OS burden and HRQoL. Design A cross-sectional study in 2017. Setting All long-term care wards in Helsinki, Finland. Participants 2401 older residents (74% females, mean age 83.9). Measurements Nurses assessed the residents and completed questionnaires on the participants' demographics, functional status, diagnoses, OS and eating habits. Nutritional status was assessed using the Mini Nutritional Assessment (MNA) and HRQoL with a 15-dimensional instrument (15D). Results Of the residents, 25.4% had one OS and 16.6% two or three OS. OS burden was associated linearly with poorer cognitive and physical functioning and a higher number of comorbidities, edentulousness without dentures, and less frequent teeth brushing/denture cleaning. OS burden was also associated with malnutrition, lower BMI and eating less during main meals. In the multivariate analyses adjusted for various confounding factors, a higher number of OS was associated with lower HRQoL. OS burden correlated with nearly all dimensions of HRQoL. Conclusion Oral symptoms are associated with generic HRQoL. Therefore, OS should be regularly assessed and managed in daily care.Peer reviewe