Genotyping of European Toxoplasma gondii strains by a new high-resolution next-generation sequencing-based method.

PURPOSE: A new high-resolution next-generation sequencing (NGS)-based method was established to type closely related European type II Toxoplasma gondii strains. METHODS: T. gondii field isolates were collected from different parts of Europe and assessed by whole genome sequencing (WGS). In comparison to ME49 (a type II reference strain), highly polymorphic regions (HPRs) were identified, showing a considerable number of single nucleotide polymorphisms (SNPs). After confirmation by Sanger sequencing, 18 HPRs were used to design a primer panel for multiplex PCR to establish a multilocus Ion AmpliSeq typing method. Toxoplasma gondii isolates and T. gondii present in clinical samples were typed with the new method. The sensitivity of the method was tested with serially diluted reference DNA samples. RESULTS: Among type II specimens, the method could differentiate the same number of haplotypes as the reference standard, microsatellite (MS) typing. Passages of the same isolates and specimens originating from abortion outbreaks were identified as identical. In addition, seven different genotypes, two atypical and two recombinant specimens were clearly distinguished from each other by the method. Furthermore, almost all SNPs detected by the Ion AmpliSeq method corresponded to those expected based on WGS. By testing serially diluted DNA samples, the method exhibited a similar analytical sensitivity as MS typing. CONCLUSION: The new method can distinguish different T. gondii genotypes and detect intra-genotype variability among European type II T. gondii strains. Furthermore, with WGS data additional target regions can be added to the method to potentially increase typing resolution.

Pre-frail older adults show improved cognition with StayFitLonger computerized home-based training: a randomized controlled trial.

Multidomain interventions have shown tremendous potential for improving cognition in older adults. It is unclear if multidomain interventions can be delivered remotely and whether remote intervention is beneficial for older adults who are vulnerable or at risk of cognitive decline. In a 26-week multi-site, home-based, double-blind, randomized controlled trial, 120 cognitively healthy older adults (75 robust, 45 pre-frail; age range = 60-94) recruited from Switzerland, Canada, and Belgium were randomized to receive either the StayFitLonger (SFL) computerized multidomain training program or an active control intervention. Delivered on tablets, the SFL intervention combined adapted physical exercises (strength, balance, and mobility), cognitive training (divided attention, problem solving, and memory), opportunities for social and contributive interactions, and psychoeducation. The active control intervention provided basic mobilization exercises and access to video games. Cognitive outcomes were global cognition (Z-scores of attention, verbal fluency, and episodic memory for nondemented older adults; ZAVEN), memory, executive function, and processing speed. Linear mixed model analyses indicated improved performance on the ZAVEN global cognition score in the SFL group but not in the active control group. Stratified analyses by frailty status revealed improved ZAVEN global cognition and processing speed scores following SFL in the pre-frail group but not in the robust group. Overall, the study indicates that a computerized program providing a multidomain intervention at home can improve cognition in older adults. Importantly, pre-frail individuals, who are at higher risk of cognitive decline, seem to benefit more from the intervention. Trial registration: ClinicalTrials.gov, NCT037519 Registered on January 22, 2020-Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04237519 .

Facilitators and obstacles to the use of a cognitive orthosis for meal preparation within the homes of adults with a moderate to severe traumatic brain injury: Informal caregivers and health-care professionals' perspectives.

A cognitive orthosis named COOK was developed and implemented to facilitate meal preparation for adults with severe traumatic brain injury (TBI) living in an alternative housing unit. This study aimed to explore facilitators and barriers to the potential use and implementation of COOK in a new context (i.e., within the homes of people living with a TBI in the community). For this purpose, 20 stakeholders (e.g., health-care professionals, clinical coordinators, informal caregivers of individuals with TBI) were interviewed. Participants identified various potential benefits of this technology (e.g., improving independence and confidence of people with TBI) and facilitators (e.g., clinical and technical supports, helpful functionalities) that could facilitate the use and implementation of COOK within a home environment. However, numerous questions remained unanswered regarding the logistics surrounding the implementation of such technology. Thus, further studies and modifications are required to facilitate future implementation of this technology among individuals living in their own homes.

Rationale and protocol of the StayFitLonger study: a multicentre trial to measure efficacy and adherence of a home-based computerised multidomain intervention in healthy older adults.

BACKGROUND: In older adults, multidomain training that includes physical and cognitive activities has been associated with improvement of physical and cognitive health. The goal of the multisite StayFitLonger study is to assess a home-based computerised training programme, which combines physical exercises, stimulating cognitive activities and virtual coaching. METHODS: One hundred twenty-eight cognitively healthy older adults will be recruited from the community in Switzerland, Canada and Belgium. The study will comprise (1) a 26-week double-blind randomized controlled efficacy trial and (2) a 22-week pragmatic adherence sub-study. In the efficacy trial, participants will be randomly assigned to an experimental or an active control intervention. In the experimental intervention, participants will use the StayFitLonger programme, which is computerised on a tablet and provides content that combines physical activities with a focus on strength and balance, as well as divided attention, problem solving and memory training. Outcomes will be measured before and after 26 weeks of training. The primary efficacy outcome will be performance on the "Timed-Up & Go" test. Secondary outcomes will include measures of frailty, cognition, mood, fear of falling, quality of life, and activities of daily living. Age, sex, education, baseline cognition, expectation, and adherence will be used as moderators of efficacy. Following the 26-week efficacy trial, all participants will use the experimental programme meaning that participants in the control group will 'cross over' to receive the StayFitLonger programme for 22 weeks. Adherence will be measured in both groups based on dose, volume and frequency of use. In addition, participants' perception of the programme and its functionalities will be characterised through usability, acceptability and user experience. DISCUSSION: This study will determine the efficacy, adherence and participants' perception of a home-based multidomain intervention programme and its functionalities. This will allow for further development and possible commercialization of a scientifically validated training programme. TRIAL REGISTRATION: ClinicalTrials.gov , NCT04237519 Registered on January 22, 2020 - Retrospectively registered.

Operators' radiation exposure reduction during cardiac catheterization using a removable shield.

Cardiac catheterization through radial access is associated with significant ionizing radiation exposure for the operator. We aimed at evaluating whether a removable shield placed upon the patient could impact favorably on annual radiation exposure for the operator. We designed a pre-post study comparing radiation exposure in a total of five operators under standard protection procedures (first period) and after applying a removable shield (second period). Each period included all the procedures performed in 1 year. Radiation exposure was measured through three dosimeters on each operator. A total of 1610 procedures were performed during the first period, and 1670 during the second period. For each operator, Fluoroscopy Time (FT) per exam did not differ between the two periods (13.1 ± 1 vs 12.9 ± 2 min/exam, p = 0.73), whereas Dose-Area Product (DAP) per procedure was slightly higher in the second period (5.247 ± 651 vs 6.374 ± 967 mGy/cm2, p < 0.01). The use of a removable shield significantly reduced operators' radiation dose at the left bracelet (64.3 ± 13.3 µSv/exam vs 23.8 ± 6.0 µSv/exam, p = 0.003). This remained significant even after adjustment for DAP per procedure (p = 0.015) and number of operators participating to each procedure (p = 0.013), whereas no significant difference was observed for card (5.6 ± 10.5 µSv/exam vs 0.9 ± 0.3 µSv/exam, p = 0.36) and neck bands (3.3 ± 4.5 µSv/exam vs 2.0 ± 2.0 µSv/exam, p = 0.36) dosimeters. The use of a removable shield during cardiac catheterization reduces radiation exposure at the level of the operator's upper limb, whereas no difference was found for other body parts. This may help in reducing radiation exposure of operator's hand. DAP increase merits further investigation.

Rationale and protocol of the ENGAGE study: a double-blind randomized controlled preference trial using a comprehensive cohort design to measure the effect of a cognitive and leisure-based intervention in older adults with a memory complaint.

BACKGROUND: Leisure activities can be both enjoyable and cognitively stimulating, and participation in such activities has been associated with reduced age-related cognitive decline. Thus, integrating stimulating leisure activities in cognitive training programs may represent a powerful and innovative approach to promote cognition in older adults at risk of dementia. The ENGAGE study is a randomized controlled, double-blind preference trial with a comprehensive cohort design that will test the efficacy and long-term impact of an intervention that combines cognitive training and cognitively stimulating leisure activities. METHODS: One hundred and forty-four older adults with a memory complaint will be recruited in Montreal and Toronto. A particular effort will be made to reach persons with low cognitive reserve. Participants will be randomly assigned to one of two conditions: cognitive + leisure training (ENGAGE-MUSIC/SPANISH) or active control (ENGAGE-DISCOVERY). The ENGAGE-MUSIC/SPANISH training will include teaching of mnemonic and attentional control strategies, casual videogames selected to train attention, and classes in music or Spanish as a second language. The ENGAGE-DISCOVERY condition will comprise psychoeducation on cognition and the brain, low-stimulating casual videogames and documentary viewing with discussions. To retain the leisure aspect of the activities, participants will be allowed to exclude either music or Spanish at study entry if they strongly dislike one of these activities. Participants randomized to ENGAGE-MUSIC/SPANISH who did not exclude any activity will be assigned to music or Spanish based on a second random assignment. Training will be provided in 24 2-h sessions over 4 months. Outcomes will be measured at baseline, at 4-month follow-up, and at 24-month follow-up. The primary outcome will be cognitive performance on a composite measure of episodic memory (delayed recall scores for words and face-name associations) measured at baseline and at the 4-month follow-up. Secondary outcomes will include a composite measure of attention (speed of processing, inhibition, dual tasking, and shifting), psychological health, activities of daily living, and brain structure and function and long-term maintenance measured at the 24-month follow-up. Information on cognitive reserve proxies (education and lifestyle questionnaires), sex and genotype (apolipoprotein (Apo)E4, brain-derived neurotrophic factor (BDNF), and catechol-O-methyltransferase (COMT)) will be collected and considered as moderators of training efficacy. DISCUSSION: This study will test whether a program combining cognitive training with stimulating leisure activities can increase cognition and reduce cognitive decline in persons at risk of dementia. TRIAL REGISTRATION: NCT03271190 . Registered on 5 September 2017.

Smartphone for smart living: Using new technologies to cope with everyday limitations in semantic dementia.

New technologies have considerable potential to support people with semantic dementia-a form of progressive aphasia-in their everyday lives, but evidence is still sparse. The first objective of the study was to document day-to-day compensation strategies, including the use of a smartphone, in ND, a 56-year-old man with semantic dementia. The second objective was to explore if, 5 years after receiving his diagnosis, ND could still learn new smartphone functions. Results for objective 1 showed that ND had adopted a large number of compensation mechanisms in his everyday life, and expanded the use of one application he had learned 4 years earlier. Results for objective 2 showed that, with an errorless learning approach, he learnt to effectively use 10 smartphone functions. He was also able to verbalise semantic knowledge about those functions and still used 40% of them in daily life 6 months post-intervention. He particularly appreciated note-taking, and spontaneously expanded his abilities in using this function's features in order to reduce his semantic difficulties. This study shows the potential of new mobile technologies for semantic dementia, how they can be adapted and modified as the disease progresses, and how some patients can creatively use external technological aids.

Relying on procedural memory to enhance independence in daily living activities: Smartphone use in a case of semantic dementia.

Relying on procedural memory is a promising approach for interventions that address the cognitive difficulties found in semantic dementia. The aim of this study was to determine if procedural memory could be used to optimise learning of relevant smartphone functions in MH, a 55-year-old man with semantic dementia. The impact of learning to use specific smartphone applications, which display concepts and their semantic characteristics, on relearning useful significant concepts, was also explored in MH. This patient, who showed no deficits in procedural learning on a serial reaction time paradigm, was able to learn manipulations related to 15 smartphone functions although, because of his deficit in word comprehension, he generally needed verbal cues to clarify which functions he was asked to perform. Six months after the end of the intervention, he was still using 8 of the 15 functions regularly. However, repeated exposure to concepts through the use of two applications did not improve naming or retrieval of semantic attributes. This study showed the potential of relying on procedural memory to optimise learning of new technologies in the ecological rehabilitation of semantic dementia.

Multiplex PCR for detection of virulence markers of Vibrio vulnificus.

UNLABELLED: Vibrio vulnificus is a Gram-negative pathogen found in coastal and estuarine waters worldwide that can cause life threatening diseases. Characterization of the vcg (virulence correlated gene) or 16S rRNA alleles is used to distinguish virulent (clinical (C)-type) from presumably avirulent (environmental (E)-type) strains. However, some studies reported a significant number of clinical strains belonging to the E-type. In recent years more potential virulence markers have been identified, that are useful for the identification of potentially pathogenic isolates of the E-type. In this study, we successfully combined detection of pathogenicity region XII, nanA and a mannitol fermentation operon with the virulence associated alleles of the 16S rRNA and vcg genes in one multiplex PCR. Additionally, toxR primers for species confirmation and internal amplification control were included. Validation of multiplex amplification was performed with a total of 132 bacterial strains, including V. vulnificus (n = 71), other Vibrionaceae (n = 50) and non-Vibrio isolates (n = 11). Multiplex PCR showed reliable amplification of four of the five virulence markers with a high sensitivity and specificity. Amplification of the 16S rRNA type B allele was not completely reliable with conventional PCR assays, however, the positive predictive value of this marker was 100 %. SIGNIFICANCE AND IMPACT OF THE STUDY: A multiplex PCR for simultaneous detection and characterization of potentially virulent strains of Vibrio vulnificus was developed and validated. Monitoring programs will benefit from this cost and time effective method when screening large strain collections. Application of the multiplex PCR simplifies determination of risks emanating from V. vulnificus in recreational waters or mussel primary production.

Genetic and phenotypic analysis of Vibrio cholerae non-O1, non-O139 isolated from German and Austrian patients.

Vibrio cholerae belonging to the non-O1, non-O139 serogroups are present in the coastal waters of Germany and in some German and Austrian lakes. These bacteria can cause gastroenteritis and extraintestinal infections, and are transmitted through contaminated food and water. However, non-O1, non-O139 V. cholerae infections are rare in Germany. We studied 18 strains from German and Austrian patients with diarrhea or local infections for their virulence-associated genotype and phenotype to assess their potential for infectivity in anticipation of possible climatic changes that could enhance the transmission of these pathogens. The strains were examined for the presence of genes encoding cholera toxin and toxin-coregulated pilus (TCP), as well as other virulence-associated factors or markers, including hemolysins, repeats-in-toxin (RTX) toxins, Vibrio seventh pandemic islands VSP-1 and VSP-2, and the type III secretion system (TTSS). Phenotypic assays for hemolysin activity, serum resistance, and biofilm formation were also performed. A dendrogram generated by incorporating the results of these analyses revealed genetic differences of the strains correlating with their clinical origin. Non-O1, non-O139 strains from diarrheal patients possessed the TTSS and/or the multifunctional autoprocessing repeats-in-toxin (MARTX) toxin, which were not found in the strains from ear or wound infections. Routine matrix-assisted laser desorption/ionization (MALDI-TOF) mass spectrometry (MS) analysis of all strains provided reliable identification of the species but failed to differentiate between strains or clusters. The results of this study indicate the need for continued surveillance of V. cholerae non-O1, non-O139 in Germany, in view of the predicted increase in the prevalence of Vibrio spp. due to the rise in surface water temperatures.

Prosthetic correction.

Fitting a comfortable and cosmetically attractive prosthetic device is a challenge, one which many people in the ophthalmic field seem to hesitate to undertake. Based on my experience, I provide practical guidelines for this important task. Artificial eyes which are required after enucleation are not discussed, but contact lenses and shells for disfigured blind or seeing eyes are.

Contact lenses in the case of the very young child.

Many hereditary congenital and infectious ocular conditions affecting young infants and children if diagnosed early enough are no longer untreatable. Special contact lenses are now available and have been specifically developed to prevent blindness, amblyopia and establish binocular vision. Different types of contact lenses and materials from which they may be fashioned are discussed and clinical considerations outlined for treatment. Visual rehabilitation of a high order is possible with proper understanding of the particular problems affecting babies at birth, later and in early infancy.

The early management of baby with Hallermann-Steriff-Francois Syndrome.

The importance of early management of sight in a baby with Hallermann-Streiff-Francois syndrome is described. Surgery for bilateral cataract was performed. Special postcataract contact lenses were fitted.