Investigating the outcomes of virus coinfection within and across host species.

Interactions between coinfecting pathogens have the potential to alter the course of infection and can act as a source of phenotypic variation in susceptibility between hosts. This phenotypic variation may influence the evolution of host-pathogen interactions within host species and interfere with patterns in the outcomes of infection across host species. Here, we examine experimental coinfections of two Cripaviruses-Cricket Paralysis Virus (CrPV), and Drosophila C Virus (DCV)-across a panel of 25 Drosophila melanogaster inbred lines and 47 Drosophilidae host species. We find that interactions between these viruses alter viral loads across D. melanogaster genotypes, with a ~3 fold increase in the viral load of DCV and a ~2.5 fold decrease in CrPV in coinfection compared to single infection, but we find little evidence of a host genetic basis for these effects. Across host species, we find no evidence of systematic changes in susceptibility during coinfection, with no interaction between DCV and CrPV detected in the majority of host species. These results suggest that phenotypic variation in coinfection interactions within host species can occur independently of natural host genetic variation in susceptibility, and that patterns of susceptibility across host species to single infections can be robust to the added complexity of coinfection.

Transcriptomic Analysis of Glycosylation and Neuroregulatory Pathways in Rodent Models in Response to Psychedelic Molecules.

The potential for psychedelic molecules in impacting cognitive flexibility has long been supported and acknowledged across scientific reports. In the current study, an approach leveraging knowledge-based gene-set information analysis has been adopted to explore the potential impact of psychedelic molecules on both glycosylation, (a post-translational modifications (PTM)) and on neuro-regulatory pathways. Though limitations and restrictions rise from the scarcity of publicly available 'omics' data, targeted analysis enabled us to identify a number of key glycogenes (Hexb, Hs6st2, Col9a2, B3gat2, Mgat5, Bgn) involved the structural organization of extracellular matrix and neuroprotective factors (Kl, Pomc, Oxt, Gal, Avp, Cartpt) which play vital roles in neuron protection, development as well as synaptic stability. In response to psychedelic molecules, we found that these genes and associated pathways are transcriptional altered in rodent models. The approach used indicates the potential to exploit existing datasets for hypothesis generation and testing for the molecular processes which play a role in the physiological response to psychedelic molecule effects. These reported findings, which focused on alterations in glycogenes and neuro-regulatory factors may provide a novel range of biomarkers to track the beneficial, as well as potential toxicological effects of psychedelic molecules.

AccessLab: Workshops to broaden access to scientific research.

AccessLabs are workshops with two simultaneous motivations, achieved through direct citizen-scientist pairings: (1) to decentralise research skills so that a broader range of people are able to access/use scientific research, and (2) to expose science researchers to the difficulties of using their research as an outsider, creating new open access advocates. Five trial AccessLabs have taken place for policy makers, media/journalists, marine sector participants, community groups, and artists. The act of pairing science academics with local community members helps build understanding and trust between groups at a time when this relationship appears to be under increasing threat from different political and economic currents in society. Here, we outline the workshop motivations, format, and evaluation, with the aim that others can build on the methods developed.

Viral susceptibility across host species is largely independent of dietary protein to carbohydrate ratios.

The likelihood of a successful host shift of a parasite to a novel host species can be influenced by environmental factors that can act on both the host and parasite. Changes in nutritional resource availability have been shown to alter pathogen susceptibility and the outcome of infection in a range of systems. Here, we examined how dietary protein to carbohydrate altered susceptibility in a large cross-infection experiment. We infected 27 species of Drosophilidae with an RNA virus on three food types of differing protein to carbohydrate ratios. We then measured how viral load and mortality across species was affected by changes in diet. We found that changes in the protein:carbohydrate in the diet did not alter the outcomes of infection, with strong positive inter-species correlations in both viral load and mortality across diets, suggesting no species-by-diet interaction. Mortality and viral load were strongly positively correlated, and this association was consistent across diets. This suggests changes in diet may give consistent outcomes across host species, and may not be universally important in determining host susceptibility to pathogens.

Reliability of phenotype estimation and extended classification of ancestry using decedent samples.

The Illumina® MiSeq FGx™, in conjunction with the ForenSeq™ DNA Signature Prep kit, produces genotypes of the CODIS-required short tandem repeats and provides phenotype and biogeographical ancestry estimations via phenotype-informative and ancestry-informative markers, respectively. Although both markers have been validated for use in forensic biology, there is little data to determine the practical utility of these estimations to assist in identifying missing persons using decedent casework samples. The accuracy and utility of phenotypic and ancestral estimations were investigated for 300 samples received by the Los Angeles County Department of Medical Examiner-Coroner. piSNP genotypes were translated into hair and eye colors using the Forenseq™ Universal Analysis Software (UAS) on the MiSeq FGx™ and the HIrisPlex System, and statistical accuracy was evaluated in context with the reported decedent characteristics. Similarly, estimates of each decedent's biogeographical ancestry were compared to assess the efficacy of these markers to predict ancestry correctly. The average UAS and the HIrisPlex system prediction accuracy for brown and blue eyes were 95.3% and 96.2%, respectively. Intermediate eye color could not be predicted with high accuracy using either system. Other than the black hair phenotype reporting an accuracy that exceeded 90% using either system, hair color was also too variable to be predicted with high accuracy. The FROG-kb database distinguishes decedents adequately beyond the Asian, African, European, and Admixed American global ancestries provided by the MiSeq FGx™ UAS PCA plots. FROG-kb correctly identified Middle Eastern, Pacific Islander, Latin American, or Jewish ancestries with accuracies of 70.0%, 81.8%, 73.8%, and 86.7%, respectively.

DNA methylation of decedent blood samples to estimate the chronological age of human remains.

Chronological age estimation may offer valuable investigative leads in human identification cases. Bisulfite pyrosequencing analysis of single CpG sites on five genes (KLF14, ELOVL2, C1orf132, TRIM59, and FHL2) was performed on 264 postmortem blood samples from individuals aged 3 months to 93 years. The goals were to develop age prediction models based on the correlation between the methylation profile and chronological age and to assess the accuracy of the prediction. Linear regression between methylation levels and age at each CpG site revealed that the five markers show a statistically significant correlation with age. The methylation data from a training set of 160 postmortem blood samples were used to develop an age prediction model with a correlation coefficient of 0.65, explaining 73.1% of age variation, with a mean absolute deviation from the chronological age of 7.60 years. The accuracy of the model was evaluated with a test set of 72 samples producing a mean absolute deviation of 7.42 years. The training and test sets were also categorized by specific age groups to assess accuracy and deviation from chronological age. The data for both sets revealed a lower prediction potential as an individual increases in age, particularly for the age categories above 50 years.

Changes in temperature alter the potential outcomes of virus host shifts.

Host shifts-where a pathogen jumps between different host species-are an important source of emerging infectious disease. With on-going climate change there is an increasing need to understand the effect changes in temperature may have on emerging infectious disease. We investigated whether species' susceptibilities change with temperature and ask if susceptibility is greatest at different temperatures in different species. We infected 45 species of Drosophilidae with an RNA virus and measured how viral load changes with temperature. We found the host phylogeny explained a large proportion of the variation in viral load at each temperature, with strong phylogenetic correlations between viral loads across temperature. The variance in viral load increased with temperature, while the mean viral load did not. This suggests that as temperature increases the most susceptible species become more susceptible, and the least susceptible less so. We found no significant relationship between a species' susceptibility across temperatures, and proxies for thermal optima (critical thermal maximum and minimum or basal metabolic rate). These results suggest that whilst the rank order of species susceptibilities may remain the same with changes in temperature, some species may become more susceptible to a novel pathogen, and others less so.

Resource quality determines the evolution of resistance and its genetic basis.

Parasites impose strong selection on their hosts, but the level of any evolved resistance may be constrained by the availability of resources. However, studies identifying the genomic basis of such resource-mediated selection are rare, particularly in nonmodel organisms. Here, we investigated the role of nutrition in the evolution of resistance to a DNA virus (PiGV), and any associated trade-offs in a lepidopteran pest species (Plodia interpunctella). Through selection experiments and whole-genome resequencing, we identify genetic markers of resistance that vary between the nutritional environments during selection. We do not find consistent evolution of resistance in the presence of virus but rather see substantial variation among replicate populations. Resistance in a low-nutrition environment is negatively correlated with growth rate, consistent with an established trade-off between immunity and development, but this relationship is highly context dependent. Whole-genome resequencing of the host shows that resistance mechanisms are likely to be highly polygenic and although the underlying genetic architecture may differ between high and low-nutrition environments, similar mechanisms are commonly used. As a whole, our results emphasize the importance of the resource environment on influencing the evolution of resistance.

The target of selection matters: An established resistance-development-time negative genetic trade-off is not found when selecting on development time.

Trade-offs are fundamental to evolutionary outcomes and play a central role in eco-evolutionary theory. They are often examined by experimentally selecting on one life-history trait and looking for negative correlations in other traits. For example, populations of the moth Plodia interpunctella selected to resist viral infection show a life-history cost with longer development times. However, we rarely examine whether the detection of such negative genetic correlations depends on the trait on which we select. Here, we examine a well-characterized negative genotypic trade-off between development time and resistance to viral infection in the moth Plodia interpunctella and test whether selection on a phenotype known to be a cost of resistance (longer development time) leads to the predicted correlated increase in resistance. If there is tight pleiotropic relationship between genes that determine development time and resistance underpinning this trade-off, we might expect increased resistance when we select on longer development time. However, we show that selecting for longer development time in this system selects for reduced resistance when compared to selection for shorter development time. This shows how phenotypes typically characterized by a trade-off can deviate from that trade-off relationship, and suggests little genetic linkage between the genes governing viral resistance and those that determine response to selection on the key life-history trait. Our results are important for both selection strategies in applied biological systems and for evolutionary modelling of host-parasite interactions.

Identification of polygonal barrel rifling characteristics in bullet wipe residue deposited on textiles.

This study examines the potential formation of land and groove rifling characteristics in bullet wipe residue deposited on textile fabrics. Textile fabrics comprised of natural, synthetic, semi-synthetic, and blended fibers were damaged either individually or as triplicate layers using two different ammunition types: Winchester WinClean 115 grain 9 mm Luger BEB and Speer 115 grain 9 mm Luger TMJ. Each ammunition type was fired in triplicate using a 9 mm Luger caliber Glock 17 at a distance of approximately 6 feet at normal incidence to the target. The bullet hole was first observed under a stereomicroscope, photographed under tungsten and infrared illumination to observe chemical residues, and subsequently tested for the presence of copper and lead residues. The preliminary chemical results show that land and groove rifling class characteristics can form in bullet wipe residue deposited on textile fabrics. The composition of the fabric, cleanliness of the interior surface of the firearm barrel, presence of intervening layers of fabric, and the bullet type are all potential contributing factors to the detection of rifling characteristics.

Reverse shoulder arthroplasty with and without latissimus and teres major transfer for patients with combined loss of elevation and external rotation: a prospective, randomized investigation.

BACKGROUND: Latissimus dorsi (LD) and teres major (TM) tendon transfers have been performed concurrently with reverse shoulder arthroplasty (RSA) to address combined loss of elevation and external rotation (CLEER) at the shoulder. The purpose of this prospective, randomized study was to compare RSA with and without LD-TM tendon transfer in patients with rotator cuff tear arthropathy and CLEER. METHODS: Patients undergoing RSA for rotator cuff tear arthropathy who also had CLEER were randomized into the group undergoing RSA with tendon transfer or the group undergoing RSA without tendon transfer. The primary outcome measure was the Activities of Daily Living and External Rotation (ADLER) score 2 years after surgery. Secondary outcome measures included the Disabilities of the Arm, Shoulder and Hand (DASH) score, American Shoulder and Elbow Surgeons (ASES) score, and Simple Shoulder Test (SST) score. RESULTS: Both the treatment and control groups showed significant improvements in the ability to perform activities of daily living requiring active external rotation measured by the ADLER score postoperatively. No significant difference in the ADLER, DASH, ASES, or SST score was found between the 2 groups at final follow-up. Resolution of the Hornblower sign occurred postoperatively in 58.3% of patients in the control group and 73.3% of those in the treatment group. CONCLUSION: No differences in the ADLER, DASH, ASES, and SST results were found between patients treated with RSA alone and those treated with RSA with LD-TM transfer for rotator cuff tear arthropathy and CLEER. Both groups showed significant improvements in all clinical outcome assessments including functional tasks that require active external rotation.

Low-Frequency Repetitive Transcranial Magnetic Stimulation to Right Parietal Cortex Disrupts Perception of Briefly Presented Stimuli.

Right parietal cortex has recently been linked to the temporal resolution of attention. We therefore sought to investigate whether disruption to right parietal cortex would affect attention to visual stimuli presented for brief durations. Participants performed a visual discrimination task before and after 10 minutes repetitive transcranial magnetic stimulation (1 Hz) to right or central parietal cortex as well as 20 minutes after the second block of trials. Participants reported the spatial frequency of a masked Gabor patch presented for a brief duration of 60, 120, or 240 ms. We calculated error magnitudes by comparing accuracy to a guessing model. We then compared error magnitudes to blocks with no stimulation, producing a measure of baselined performance. Baselined performance was poorer at longer stimulus durations after right parietal than central parietal stimulation, suggesting that right parietal cortex is involved in attention to briefly presented stimuli, particularly in situations where rapid accumulation of visual evidence is needed.

Opioid-free shoulder arthroplasty: a prospective study of a novel clinical care pathway.

BACKGROUND: Opioid therapy has been a cornerstone of perioperative pain control for decades in the United States, despite our increased understanding of the morbidity and mortality linked to opioids. The purpose of this study is to explore the safety, efficacy, and feasibility of an entirely opioid-free perioperative pathway in patients undergoing elective shoulder arthroplasty. METHODS: Thirty-five patients undergoing elective total shoulder arthroplasty with a mean age of 71 (range, 50-87) years elected into a comprehensive opioid-free, multimodal pain management protocol. Opioid use was completely eliminated for all points in the perioperative period including during regional and general anesthesia. Data were collected regarding patient-reported pain, opioid consumption in the perioperative period, postoperative delirium, nausea, constipation, and falls. RESULTS: Pain level at the primary outcome point of 24 hours or discharge was rated at 2.5 on the numeric rating scale. Stable, low pain scores were demonstrated at all time points postoperatively. Low rates of nausea, falls, and constipation were reported. Only 1 patient required "rescue" opioid medications during the in-patient stay, and an additional patient was given a low-dose opioid prescription at the 2-week postoperative appointment. CONCLUSIONS: An opioid-free, multimodal pain management pathway is a safe and effective option in properly selected patients undergoing shoulder arthroplasty with a very low risk of requiring rescue opioids. This study is the first such study to present a surgical protocol entirely free of opioids at all portions of the patient care pathway.

The Effect of a Low-Cost Body Weight-Supported Treadmill Trainer on Walking Speed and Joint Motion.

Background and Objectives: Gait training with body weight-support has been shown to improve the walking speed of individuals with movement disorders. The AccesSportAmerica Gait Trainer is a low-cost, pre-market gait rehabilitation device that alters the stride characteristics of participants walking on a standard treadmill. The purpose of this study was to examine the biomechanical outcomes that training on this device has for people with brain injuries that affect motor functioning. It was hypothesized that there would be an increase in walking speed post-intervention, and that there would be an increase in step length and joint range-of-motion. Materials and Methods: An intervention study was conducted with 11 people with ambulatory difficulty caused by post-stroke hemiparesis (n = 7), traumatic brain injury (n = 3), and cerebral palsy (n = 1). The average time using the AccesSportAmerica Gait Trainer was 34.5 (SD = 6.0) minutes per session for 36.9 (SD = 21.8) sessions. Gait speed, step length and time, and joint flexion were measured during the 10 Meter Walk Test. Results: From pre- to post-intervention, there was a mean increase in walking speed of 0.19 m/s (SD = 0.06, p = 0.016, d = 0.40) and a decrease in step time of both affected and unaffected legs (affected: p = 0.011, d = 0.37; unaffected: p = 0.004, d = 0.67). There was no significant change in stride length or joint angles. Conclusions: The AccesSportAmerica Gait Trainer has the potential to improve the walking speed of people with ambulatory difficulty.

[Lost in transition: crossed looks on the 18-25 years old]. / Lost in transition :regards croisés sur les 18-25 ans.

Social determinants have an impact on patients' access to care and on their health at any age. Young adults are a population at risk of being affected in their health because of a lack of social protection or unavailability of healthcare system adapted to their specific needs. Primary healthcare providers need to identify that window of social vulnerability in order to avoid a discontinuation in healthcare and to prevent biopsychosocial co-morbidity leading to ongoing consequences on their health.

L'impact des déterminants sociaux sur l'accès aux soins des patients et sur leur santé concerne toutes les catégories d'âge. Les jeunes adultes sont une population particulièrement à risque d'être pénalisés dans leur santé par un défaut de protection sociale ou de réseau de soins adapté à leurs besoins spécifiques. Le médecin de premier recours, pédiatre ou généraliste, doit identifier cette fenêtre de vulnérabilité sociale afin d'éviter une rupture de soins et prévenir les comorbidités biopsychosociales ayant des conséquences durables sur la santé.

DNA Gyrase Is the Target for the Quinolone Drug Ciprofloxacin in Arabidopsis thaliana.

The Arabidopsis thaliana genome contains four genes that were originally annotated as potentially encoding DNA gyrase: ATGYRA, ATGYRB1, ATGYRB2, and ATGYRB3. Although we subsequently showed that ATGYRB3 does not encode a gyrase subunit, the other three genes potentially encode subunits of a plant gyrase. We also showed evidence for the existence of supercoiling activity in A. thaliana and that the plant is sensitive to quinolone and aminocoumarin antibiotics, compounds that target DNA gyrase in bacteria. However, it was not possible at that time to show whether the A. thaliana genes encoded an active gyrase enzyme, nor whether that enzyme is indeed the target for the quinolone and aminocoumarin antibiotics. Here we show that an A. thaliana mutant resistant to the quinolone drug ciprofloxacin has a point mutation in ATGYRA. Moreover we show that, as in bacteria, the quinolone-sensitive (wild-type) allele is dominant to the resistant gene. Further we have heterologously expressed ATGYRA and ATGYRB2 in a baculovirus expression system and shown supercoiling activity of the partially purified enzyme. Expression/purification of the quinolone-resistant A. thaliana gyrase yields active enzyme that is resistant to ciprofloxacin. Taken together these experiments now show unequivocally that A. thaliana encodes an organelle-targeted DNA gyrase that is the target of the quinolone drug ciprofloxacin; this has important consequences for plant physiology and the development of herbicides.

Forensic intelligence applied to questioned document analysis: A model and its application against organized crime.

The capability of forensic sciences to fight crime, especially against organized criminal groups, becomes relevant in the recent economic downturn and the war on terrorism. In view of these societal challenges, the methods of combating crime should experience critical changes in order to improve the effectiveness and efficiency of the current resources available. It is obvious that authorities have serious difficulties combating criminal groups of transnational nature. These are characterized as well structured organizations with international connections, abundant financial resources and comprised of members with significant and diverse expertise. One common practice among organized criminal groups is the use of forged documents that allow for the commission of illegal cross-border activities. Law enforcement can target these movements to identify counterfeits and establish links between these groups. Information on document falsification can become relevant to generate forensic intelligence and to design new strategies against criminal activities of this nature and magnitude. This article discusses a methodology for improving the development of forensic intelligence in the discipline of questioned document analysis. More specifically, it focuses on document forgeries and falsification types used by criminal groups. It also describes the structure of international criminal organizations that use document counterfeits as means to conduct unlawful activities. The model presented is partially based on practical applications of the system that have resulted in satisfactory outcomes in our laboratory.

ACTIVE SURVEILLANCE FOR PAPILLARY THYROID MICROCARCINOMA: NEW CHALLENGES AND OPPORTUNITIES FOR THE HEALTH CARE SYSTEM.

OBJECTIVE: The dramatic increase in papillary thyroid carcinoma (PTC) is primarily a result of early diagnosis of small cancers. Active surveillance is a promising management strategy for papillary thyroid microcarcinomas (PTMCs). However, as this management strategy gains traction in the U.S., it is imperative that patients and clinicians be properly educated, patients be followed for life, and appropriate tools be identified to implement the strategy. METHODS: We review previous active surveillance studies and the parameters used to identify patients who are good candidates for active surveillance. We also review some of the challenges to implementing active surveillance protocols in the U.S. and discuss how these might be addressed. RESULTS: Trials of active surveillance support nonsurgical management as a viable and safe management strategy. However, numerous challenges exist, including the need for adherence to protocols, education of patients and physicians, and awareness of the impact of this strategy on patient psychology and quality of life. The Thyroid Cancer Care Collaborative (TCCC) is a portable record keeping system that can manage a mobile patient population undergoing active surveillance. CONCLUSION: With proper patient selection, organization, and patient support, active surveillance has the potential to be a long-term management strategy for select patients with PTMC. In order to address the challenges and opportunities for this approach to be successfully implemented in the U.S., it will be necessary to consider psychological and quality of life, cultural differences, and the patient's clinical status.