Longitudinal Microbiome Changes in Supragingival Biofilm Transcriptomes Induced by Orthodontics.

INTRODUCTION: Common oral diseases are known to be associated with dysbiotic shifts in the supragingival microbiome, yet most oral microbiome associations with clinical end points emanate from cross-sectional studies. Orthodontic treatment is an elective procedure that can be exploited to prospectively examine clinically relevant longitudinal changes in the composition and function of the supragingival microbiome. METHODS: A longitudinal cohort study was conducted among 24 adolescent orthodontic patients who underwent saliva and plaque sampling and clinical examinations at time points: before fixed appliance bonding and at 1, 6, and 12 wk thereafter. Clinical indices included bleeding on probing (BOP), mean gingival index (GI), probing depths (PDs), and plaque index (PI). To study the biologically (i.e., transcriptionally) active microbial communities, RNA was extracted from plaque and saliva for RNA sequencing and microbiome bioinformatics analysis. Longitudinal changes in microbiome beta diversity were examined using PERMANOVA tests, and the relative abundance of microbial taxa was measured using Kruskal-Wallis tests, Wilcoxon rank-sum tests, and negative binomial and zero-inflated mixed models. RESULTS: Clinical measures of oral health deteriorated over time-the proportion of sites with GI and PI ≥1 increased by over 70% between prebonding and 12 wk postbonding while the proportion of sites with PD ≥4 mm increased 2.5-fold. Streptococcus sanguinis, a health-associated species that antagonizes cariogenic pathogens, showed a lasting decrease in relative abundance during orthodontic treatment. Contrarily, caries- and periodontal disease-associated taxa, including Selenomonas sputigena, Leptotrichia wadei, and Lachnoanaerobaculum saburreum, increased in abundance after bonding. Relative abundances of Stomatobaculum longum and Mogibacterium diversum in prebonding saliva predicted elevated BOP 12 wk postbonding, whereas Neisseria subflava was associated with lower BOP. CONCLUSIONS: This study offers insights into longitudinal community and species-specific changes in the supragingival microbiome transcriptome during fixed orthodontic treatment, advancing our understanding of microbial dysbioses and identifying targets of future health-promoting clinical investigations. KNOWLEDGE TRANSFER STATEMENT: Bonding braces was associated with subsequent changes in the oral microbiome characterized by increases in disease-associated species, decreases in health-associated species, and worsened clinical measures of oral health.

Selective Visualization of Live Intestinal Parasites in Stool Specimens Without Purification.

PURPOSE: Microscopic observation of live parasites in the stool is an important diagnostic tool in human and veterinary medicine. Because of the presence of large amounts of contaminating organic matter, microscopic analysis must be preceded by time-consuming pre-purification steps. Transmission-through-dye (TTD) optical microscopy obviates this problem. The purpose of this study is to illustrate the use of TTD for the analysis of stool samples. METHODS: TTD imaging is based on the exclusion of a strongly absorbing dye by living cells. A food colorant Acid Blue 9 (AB9) is added to a fecal suspension, and the sample is observed under transmitted illumination through a 630 nm bandpass filter. AB9 strongly absorbs red light, and it comes out of the sample significantly attenuated. However, if a viable cell of any origin is present in the sample, it excludes the dye and reduces the depth of the light-absorbing layer. RESULTS: Live cells or eggs appear bright red on a dark background, while most of the organic contaminants remain dark. The method has been demonstrated on schistosomes, hookworms, giardia, yeast, and other organisms. CONCLUSION: TTD dramatically increases the visibility of live parasites and permits their direct observation in a fecal suspension. TTD can be enabled on any microscope by simply adding a red filter.

A double-blind, 377-subject randomized study identifies Ruminococcus, Coprococcus, Christensenella, and Collinsella as long-term potential key players in the modulation of the gut microbiome of lactose intolerant individuals by galacto-oligosaccharides.

Background. Our recent publication (Chey et al., Nutrients 2020) showed that a 30-day administration of pure galacto-oligosaccharides (GOS) significantly reduced symptoms and altered the fecal microbiome in patients with lactose intolerance (LI). Results. In this addendum, we performed an in-depth analysis of the fecal microbiome of the 377 LI patients randomized to one of two GOS doses (Low, 10-15 grams/day or High, 15-20 grams/day), or placebo in a multi-center, double-blinded, placebo-controlled trial. Sequencing of 16S rRNA amplicons was done on GOS or placebo groups at weeks zero (baseline), four (end of treatment), nine, 16 and 22. Taxa impacted by treatment and subsequent dairy consumption included lactose-fermenting species of Bifidobacterium, Lactobacillus, Lactococcus, and Streptococcus. Increased secondary fermentation microorganisms included Coprococcus and Ruminococcus species, Blautia producta, and Methanobrevibacterium. Finally, tertiary fermenters that use acetate to generate butyrate were also increased, including Faecalibacterium prausnitzii, Roseburia faecis, and C. eutactus. Conclusions. Results confirmed and expanded data on GOS microbiome modulation in LI individuals. Microbiome analysis at 16 and 22 weeks after treatment further suggested relatively long-term benefits when individuals continued consumption of dairy products.

Novel, bilateral, two-bellied muscles span the extensor forearm, thenar eminence to insert on the proximal phalanx of the thumb: clinical and embryological significance.

Muscle and tendon variations in the forearm, wrist and hand are commonly reported in the anatomical and surgical literature. They are frequently the source of inflammatory conditions such as de Quervain's tenosynovitis or carpal tunnel syndrome. During academic dissection, a cadaver presented with bilateral, additional muscles running parallel to the abductor pollicis longus muscles (APL) in the extensor compartment of the forearm. Both additional muscles had two bellies, one proximal and one distal, with an intervening tendon. The proximal bellies were separate and distinct from the adjacent APLs. The tendons traversed the first dorsal compartments with the tendons of the APLs and the extensor pollicis brevis muscles (EPB). The distal bellies lay adjacent to the abductor pollicis brevis (APB) muscles in the thenar compartments, and inserted onto the volar base of the proximal phalanges of the thumbs. Following a thorough search of the literature, we determined that these additional muscles constitute a previously unreported variation. This report details the variation, compares it with other reported variations, presents the related embryology, and reviews the significance of this variation as it relates to inflammatory conditions and surgical procedures.

Development of Subject-Specific Proximal Femur Finite Element Models Of Older Adults with Obesity to Evaluate the Effects of Weight Loss on Bone Strength.

STUDY BACKGROUND: Recommendation of intentional weight loss in older adults remains controversial, due in part to the loss of bone mineral density (BMD) known to accompany weight loss. While finite element (FE) models have been used to assess bone strength, these methods have not been used to study the effects of weight loss. The purpose of this study is to develop subject-specific FE models of the proximal femur and study the effect of intentional weight loss on bone strength. METHODS: Computed tomography (CT) scans of the proximal femur of 25 overweight and obese (mean BMI=29.7 ± 4.0 kg/m2), older adults (mean age=65.6 ± 4.1 years) undergoing an 18-month intentional weight loss intervention were obtained at baseline and post-intervention. Measures of volumetric BMD (vBMD) and variable cortical thickness were derived from each subject CT scan and directly mapped to baseline and post-intervention models. Subject-specific FE models were developed using morphing techniques. Bone strength was estimated through simulation of a single-limb stance and sideways fall configuration. RESULTS: After weight loss intervention, there were significant decreases from baseline to 18 months in vBMD (total hip: -0.024 ± 0.013 g/cm3; femoral neck: -0.012 ± 0.014 g/cm3), cortical thickness (total hip: -0.044 ± 0.032 mm; femoral neck: -0.026 ± 0.039 mm), and estimated strength (stance: -0.15 ± 0.12 kN; fall: -0.04 ± 0.06 kN). Adjusting for baseline bone measures, body mass, and gender, correlations were found between weight change and change in total hip and femoral neck cortical thickness (all p<0.05). For every 1 kilogram of body mass lost cortical thickness in the total hip and femoral neck decreased by 0.003 mm and 0.004 mm, respectively. No significant correlations were present for the vBMD or strength data. CONCLUSION: The developed subject-specific FE models could be used to better understand the effects of intentional weight loss on bone health.

Can we predict which dysplastic hips will require acetabular augmentation during total hip arthroplasty based on pre-operative radiographs?

AIMS: The purpose of this study was to evaluate whether an innovative templating technique could predict the need for acetabular augmentation during primary total hip arthroplasty for patients with dysplastic hips. PATIENTS AND METHODS: We developed a simple templating technique to estimate acetabular component coverage at total hip arthroplasty, the True Cup: False Cup (TC:FC) ratio. We reviewed all patients with dysplastic hips who underwent primary total hip arthroplasty between 2005 and 2012. Traditional radiological methods of assessing the degree of acetabular dysplasia (Sharp's angle, Tönnis angle, centre-edge angle) as well as the TC:FC ratio were measured from the pre-operative radiographs. A comparison of augmented and non-augmented hips was undertaken to determine any difference in pre-operative radiological indices between the two cohorts. The intra- and inter-observer reliability for all radiological indices used in the study were also calculated. RESULTS: Of the 128 cases reviewed, 33 (26%) needed acetabular augmentation. We found no difference in the median Sharp's angle (p = 0.10), Tönnis angle (p = 0.28), or centre-edge angle (p = 0.07) between the two groups. A lower TC:FC ratio was observed in the augmented group compared with the non-augmented group (median = 0.66 versus 0.88, p < 0.001). Intra-observer reliability was found to be high for all radiological indices analysed (interclass correlation coefficient (ICC) > 0.7). However, inter-observer reliability was more variable and was only high for the TC: FC ratio (ICC > 0.7). CONCLUSION: The TC: FC ratio gives an accurate estimate of acetabular component coverage. It can help predict which dysplastic hips are likely to need acetabular augmentation at primary total hip arthroplasty. It has high intra- and inter-observer reliability. Cite this article: Bone Joint J 2017;99-B:445-50.

Imaging of Myocardial Fibrosis in Patients with End-Stage Renal Disease: Current Limitations and Future Possibilities.

Cardiovascular disease in patients with end-stage renal disease (ESRD) is driven by a different set of processes than in the general population. These processes lead to pathological changes in cardiac structure and function that include the development of left ventricular hypertrophy and left ventricular dilatation and the development of myocardial fibrosis. Reduction in left ventricular hypertrophy has been the established goal of many interventional trials in patients with chronic kidney disease, but a recent systematic review has questioned whether reduction of left ventricular hypertrophy improves cardiovascular mortality as previously thought. The development of novel imaging biomarkers that link to cardiovascular outcomes and that are specific to the disease processes in ESRD is therefore required. Postmortem studies of patients with ESRD on hemodialysis have shown that the extent of myocardial fibrosis is strongly linked to cardiovascular death and accurate imaging of myocardial fibrosis would be an attractive target as an imaging biomarker. In this article we will discuss the current imaging methods available to measure myocardial fibrosis in patients with ESRD, the reliability of the techniques, specific challenges and important limitations in patients with ESRD, and how to further develop the techniques we have so they are sufficiently robust for use in future clinical trials.

Outcome of elective withdrawal of anti-tumour necrosis factor-α therapy in patients with Crohn's disease in established remission.

BACKGROUND AND AIMS: Outcomes of cessation of anti-TNF therapy for Crohn's disease (CD) in clinical and/or endoscopic remission in routine clinical practice is uncertain. This study aimed to evaluate clinical outcomes and factors associated with relapse in CD patients following formal disease assessment and elective anti-TNF withdrawal. METHODS: Prospective observational study of CD patients in whom anti-TNF therapy was stopped electively after ≥12months and follow-up of ≥6months. Investigations at assessment prior to cessation included ≥1 of clinical assessment, endoscopic and/or imaging. Relapse was defined as recurrent symptoms of CD requiring medical or surgical therapy. RESULTS: Eighty-six patients received anti-TNF for a median duration of 23 (12-80) months for severe active luminal (70%), fistulating perianal (25.5%) and other fistulating disease (4.5%). Relapse rates at 90,180 and 365days were 4.7%, 18.6% and 36%, respectively. If anti-TNF dose escalation occurred 6months prior to withdrawal, 88% (7/8) relapsed. Based on multivariate analysis, risk factors for relapse include ileocolonic disease at diagnosis and previous anti-TNF therapy. An elevated faecal calprotectin (FC) is likely to predict relapse (p=0.02), with a PPV of 66.7% at >50µg/g. Of 36 patients who relapsed, 31 were retreated with anti-TNF, with an overall recapture rate of 93%. CONCLUSION: Relapse rates at 1year following elective withdrawal of anti-TNF are 36%, with high retreatment response rate. Predictors of relapse include ileocolonic involvement, previous anti-TNF therapy and raised FC. Endoscopic/radiologic assessment prior to cessation of therapy does not appear to predict those at lower risk of relapse.

Legacy effects of short-term intentional weight loss on total body and thigh composition in overweight and obese older adults.

OBJECTIVE: Weight regain following intentional weight loss may negatively impact body composition, accelerating fat regain and increasing risk of physical disability. The purpose of this study was to compare long-term changes in whole body and thigh composition in obese older adults who intentionally lost and then partially regained weight to obese older adults who remained weight stable. SUBJECTS/METHODS: This pilot study analyzed total body (dual-energy X-ray absorptiometry (DXA)) and thigh (computed tomography (CT)) composition data collected from 24 older (65-79 years) adults 18 months after completion of a 5-month randomized trial that compared resistance training alone (RT) with RT plus caloric restriction (RT+CR). RESULTS: Mean loss of body mass in the RT+CR group (n=13) was 7.1±2.4 kg during the 5-month intervention (74% fat mass; 26% lean mass; all P<0.01), whereas RT (n=11) remained weight stable (+0.3±1.8 kg; P=0.64). Differential group effects were observed for all DXA and CT body composition measures at 5 months (all P⩽0.01); however, by 23 months, group differences persisted only for total body (RT+CR: 81.6±10.0 kg vs RT: 88.5±14.9 kg; P=0.03) and lean (RT+CR: 50.8±9.3 kg vs RT: 54.4±12.0 kg; P<0.01) mass. All RT+CR participants regained weight from 5 to 23 months (mean gain=+4.8±2.6 kg; P<0.01). Total fat mass and all thigh fat volumes increased, whereas thigh muscle volume decreased, during the postintervention follow-up in RT+CR (all P⩽0.01). In the RT group, body mass did not change from 5 to 23 months (-0.2±0.9 kg; P=0.87). Decreased total thigh volume, driven by the loss of thigh muscle volume, were the only postintervention body composition changes observed in the RT group (both P<0.04). CONCLUSIONS: Short-term body composition benefits of an RT+CR intervention may be lost within 18 months after completion of the intervention.

Detection of Hammondia heydorni DNA in feces collected in and around an Ohio Wildlife Conservation Center.

This study aimed to identify DNA attributed to Hammondia heydorni oocysts in the feces of wild canids in and around an Ohio wildlife conservation center. Two hundred and eighty-five wild canid fecal samples were analyzed using PCR with melting curve analysis to detect coccidian DNA. Coccidia-positive samples were further subjected to H. heydorni-specific and N. caninum-specific PCR assays. Samples positive by the H. heydorni-specific assay were additionally analyzed with a PCR assay to distinguish H. heydorni from Hammondia triffittae. Coccidian DNA was detected in 51 of the 285 (17.9%) wildlife samples. H. heydorni DNA was detected in three of the coccidia-positive wildlife samples (1.1%) and N. caninum was detected in none. Determining the presence of H. heydorni in wild canids will contribute to a greater understanding of the role these hosts play in the ecology of this parasite.

Callous-unemotional traits drive reduced white-matter integrity in youths with conduct problems.

BACKGROUND: Callous-unemotional (CU) traits represent a significant risk factor for severe and persistent conduct problems in children and adolescents. Extensive neuroimaging research links CU traits to structural and functional abnormalities in the amygdala and ventromedial prefrontal cortex. In addition, adults with psychopathy (a disorder for which CU traits are a developmental precursor) exhibit reduced integrity in uncinate fasciculus, a white-matter (WM) tract that connects prefrontal and temporal regions. However, research in adolescents has not yet yielded similarly consistent findings. METHOD: We simultaneously modeled CU traits and externalizing behaviors as continuous traits, while controlling for age and IQ, in order to identify the unique relationship of each variable with WM microstructural integrity, assessed using diffusion tensor imaging. We used tract-based spatial statistics to evaluate fractional anisotropy, an index of WM integrity, in uncinate fasciculus and stria terminalis in 47 youths aged 10-17 years, of whom 26 exhibited conduct problems and varying levels of CU traits. RESULTS: Whereas both CU traits and externalizing behaviors were negatively correlated with WM integrity in bilateral uncinate fasciculus and stria terminalis/fornix, simultaneously modeling both variables revealed that these effects were driven by CU traits; the severity of externalizing behavior was not related to WM integrity after controlling for CU traits. CONCLUSIONS: These results indicate that WM abnormalities similar to those observed in adult populations with psychopathy may emerge in late childhood or early adolescence, and may be critical to understanding the social and affective deficits observed in this population.

Sarcocyst Development in Raccoons (Procyon lotor) Inoculated with Different Strains of Sarcocystis neurona Culture-Derived Merozoites.

Sarcocystis neurona is considered the major etiologic agent of equine protozoal myeloencephalitis (EPM), a neurological disease in horses. Raccoon ( Procyon lotor ) is considered the most important intermediate host in the life cycle of S. neurona in the United States; S. neurona sarcocysts do mature in raccoon muscles, and raccoons also develop clinical signs simulating EPM. The focus of this study was to determine if sarcocysts would develop in raccoons experimentally inoculated with different host-derived strains of in vitro-cultivated S. neurona merozoites. Four raccoons were inoculated with strains derived from a raccoon, a sea otter, a cat, and a horse. Raccoon tissues were fed to laboratory-raised opossums ( Didelphis virginiana ), the definitive host of S. neurona . Intestinal scraping revealed sporocysts in opossums who received muscle tissue from raccoons inoculated with the raccoon-derived or the sea otter-derived isolates. These results demonstrate that sarcocysts can mature in raccoons inoculated with in vitro-derived S. neurona merozoites. In contrast, the horse and cat-derived isolates did not produce microscopically or biologically detected sarcocysts. Immunoblot analysis revealed both antigenic and antibody differences when testing the inoculated raccoons. Immunohistochemical staining indicated differences in staining between the merozoite and sarcocyst stages. The successful infections achieved in this study indicates that the life cycle can be manipulated in the laboratory without affecting subsequent stage development, thereby allowing further purification of strains and artificial maintenance of the life cycle.

An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).

Equine protozoal myeloencephalitis (EPM) is a serious disease of horses, and its management continues to be a challenge for veterinarians. The protozoan Sarcocystis neurona is most commonly associated with EPM. S. neurona has emerged as a common cause of mortality in marine mammals, especially sea otters (Enhydra lutris). EPM-like illness has also been recorded in several other mammals, including domestic dogs and cats. This paper updates S. neurona and EPM information from the last 15 years on the advances regarding life cycle, molecular biology, epidemiology, clinical signs, diagnosis, treatment and control.

Developing the radiation protection safety culture in the UK.

In the UK, as elsewhere, there is potential to improve how radiological challenges are addressed through improvement in, or development of, a strong radiation protection (RP) safety culture. In preliminary work in the UK, two areas have been identified as having a strong influence on UK society: the healthcare and nuclear industry sectors. Each has specific challenges, but with many overlapping common factors. Other sectors will benefit from further consideration.In order to make meaningful comparisons between these two principal sectors, this paper is primarily concerned with cultural aspects of RP in the working environment and occupational exposures rather than patient doses.The healthcare sector delivers a large collective dose to patients each year, particularly for diagnostic purposes, which continues to increase. Although patient dose is not the focus, it must be recognised that collective patient dose is inevitably linked to collective occupational exposure, especially in interventional procedures.The nuclear industry faces major challenges as work moves from operations to decommissioning on many sites. This involves restarting work in the plants responsible for the much higher radiation doses of the 1960/70s, but also performing tasks that are considerably more difficult and hazardous than those original performed in these plants.Factors which influence RP safety culture in the workplace are examined, and proposals are considered for a series of actions that may lead to an improvement in RP culture with an associated reduction in dose in many work areas. These actions include methods to improve knowledge and awareness of radiation safety, plus ways to influence management and colleagues in the workplace. The exchange of knowledge about safety culture between the nuclear industry and medical areas may act to develop RP culture in both sectors, and have a wider impact in other sectors where exposures to ionising radiations can occur.

Who should answer the question: "Can I drive with this plaster cast?".

INTRODUCTION: The application of a plaster cast is known to affect driving ability, but patients continue to drive. The individuals and authorities involved in assessing driving safely include doctors, the Driver and Vehicle Licensing Agency (DVLA), police, insurance companies, and patients, but it is unclear who should take responsibility for the advice given, especially in the event of an accident. METHODS: We contacted senior plaster technicians in 348 hospitals in the UK. We recorded their responses regarding advice given to patients on driving in specific casts. Sixteen motor insurance companies and 40 police forces were also contacted in order to canvass their opinions. RESULTS: 188 technician interviews (response rate 54%) were conducted. Only 10% of respondents offered advice unprompted; an average of 48% of patients asked for advice. 88% of respondents referred patients to their motor insurance companies, and also to the DVLA (11.7%), doctor (10.6%), or police (5.9%). Only 20.2% of plaster rooms provided written information. All insurance companies would insure patients provided the doctor had not explicitly objected to driving, but there was no consensus amongst the responses received from police. In the event of an accident after the treating doctor had advised against driving, insurance companies were likely to invalidate the policy, and the police would seek penalty punishment or prosecution. CONCLUSIONS: Although doctors are not specifically trained to assess the ability of patients to drive, insurance companies and police forces place the responsibility on doctors to advise patients. Since current evidence suggests plaster casts can impair driving ability, we suggest patients should be advised not to drive. Patients accept all responsibility if they continue to drive after receiving this specific advice and understanding its implications.

False-positive buprenorphine by CEDIA in patients prescribed amisulpride or sulpiride.

Buprenorphine is a potent partial opioid agonist that is analyzed in urine to (i) monitor adherence to maintenance or detoxification therapy and (ii) detect illicit use. Buprenorphine analysis is commonly conducted on urine by immunoassay, but is subject to cross-reactivity from other drugs/drug metabolites, including morphine, codeine and dihydrocodeine. This study reports false-positive buprenorphine analysis [Thermo Fisher Scientific cloned enzyme donor immunoassay (CEDIA)] in patients who denied unauthorized buprenorphine use prior to sampling, but who had been prescribed amisulpride. In two cases, confirmatory analysis by liquid chromatography-tandem mass spectrometry was negative (<0.5 µg/L) for buprenorphine and metabolites and positive for amisulpride. Although the cross-reactivity of amisulpride and sulpiride in the CEDIA buprenorphine assay is low (estimated at 0.003 and 0.002%, respectively), it remains a significant consideration given the likely high concentrations of these compounds in urine relative to the low cutoff of the buprenorphine assay. Neither amisulpride nor sulpiride was listed as potential sources of interference on the CEDIA data sheet when this work was performed. These findings highlight the importance of confirming immunoassay-positive buprenorphine results using a more selective analytical technique.

The effect of increased overjet on the magnitude and reproducibility of smiling in adult females.

The objective of this study was to determine if increased overjet (greater than 6 mm) influences the magnitude and reproducibility of natural smile and maximal smile in Caucasian adult females. Twenty adult females with an increased overjet (6-10 mm) and 20 control adult females (overjet 2-4 mm) with no history of orthodontic treatment volunteered to participate. The mean age in the control group was 30.1 ± 6.4 years and the mean age in the test group was 31.9 ± 10.8 years. Three-dimensional stereophotogrammetric images were captured of each subject for three expressions: at rest, natural smile, and maximal smile. The images were recorded twice on two separate occasions, 6 weeks apart. Images were landmarked and a partial ordinary Procrustes superimposition was used to adjust for the differences in head posture between the same expressions. The magnitude of movement relative to the rest position, averaged over all the landmarks, was calculated and compared between the groups using analysis of variance (linear mixed-effects model); the intra- and inter-session reproducibility of both expressions was assessed. There was greater mean movement, averaged over all the landmarks, in the control group than in the increased overjet group for both natural smile and maximal smile (P = 0.0068). For these expressions, there were no statistically significant differences in reproducibility within sessions (P = 0.5403) or between sessions (P = 0.3665). Increased overjet had a statistically significant effect on the magnitude of smiling but did not influence the reproducibility of natural or maximal smile relative to controls.