Gut dysbiosis in cutaneous T-cell lymphoma is characterized by shifts in relative abundances of specific bacterial taxa and decreased diversity in more advanced disease.

BACKGROUND: Cutaneous T-cell lymphoma (CTCL) patients often suffer from recurrent skin infections and profound immune dysregulation in advanced disease. The gut microbiome has been recognized to influence cancers and cutaneous conditions; however, it has not yet been studied in CTCL. OBJECTIVES: To investigate the gut microbiome in patients with CTCL and in healthy controls. METHODS: A case-control study was conducted between January 2019 and November 2020 at Northwestern's busy multidisciplinary CTCL clinic (Chicago, Illinois, USA) utilizing 16S ribosomal RNA gene amplicon sequencing and bioinformatics analyses to characterize the microbiota present in fecal samples of CTCL patients (n = 38) and age-matched healthy controls (n = 13) from the same geographical region. RESULTS: Gut microbial α-diversity trended lower in patients with CTCL and was significantly lower in patients with advanced CTCL relative to controls (P = 0.015). No differences in ß-diversity were identified. Specific taxa were significantly reduced in patient samples; significance was determined using adjusted P-values (q-values) that accounted for a false discovery rate threshold of 0.05. Significantly reduced taxa in patient samples included the phylum Actinobacteria (q = 0.0002), classes Coriobacteriia (q = 0.002) and Actinobacteria (q = 0.03), order Coriobacteriales (q = 0.003), and genus Anaerotruncus (q = 0.01). The families Eggerthellaceae (q = 0.0007) and Lactobacillaceae (q = 0.02) were significantly reduced in patients with high skin disease burden. CONCLUSIONS: Gut dysbiosis can be seen in patients with CTCL compared to healthy controls and is pronounced in more advanced CTCL. The taxonomic shifts associated with CTCL are similar to those previously reported in atopic dermatitis and opposite those of psoriasis, suggesting microbial parallels to the immune profile and skin barrier differences between these conditions. These findings may suggest new microbial disease biomarkers and reveal a new angle for intervention.

Structures and Dynamics of Anionic Lipoprotein Nanodiscs.

Nanolipoprotein particles known as nanodiscs (NDs) have emerged as versatile and powerful tools for the stabilization of membrane proteins permitting a plethora of structural and biophysical studies. Part of their allure is their flexibility to accommodate many types of lipids and precise control of the composition. However, little is known about how variations in lipid composition impact their structures and dynamics. Herein, we investigate how the introduction of the anionic lipid POPG into POPC NDs impacts these features. Small-angle X-ray and neutron scattering (SAXS and SANS) of variable-composition NDs are complemented with molecular dynamics simulations to interrogate how increasing the concern of POPG impacts the ND shape, structure of the lipid core, and the dynamics of the popular membrane scaffold protein, MSP1D1(-). A convenient benefit of including POPG is that it eliminates D2O-induced aggregation observed in pure POPC NDs, permitting studies by SANS at multiple contrasts. SAXS and SANS data could be globally fit to a stacked elliptical cylinder model as well as an extension of the model that accounts for membrane curvature. Fitting to both models supports that the introduction of POPG results in strongly elliptical NDs; however, MD simulations predict the curvature of the membrane, thereby supporting the use of the latter model. Trends in the model-independent parameters suggest that increases in POPG reduce the conformational heterogeneity of the MSP1D1(-), which is in agreement with MD simulations that show that the incorporation of sufficient POPG suppresses disengagement of the N-terminal helix from the lipid core. These studies highlight novel structural changes in NDs in response to an anionic lipid and will inform the interpretation of future structural studies of membrane proteins embedded in NDs of mixed lipid composition.

Why don't we have more inhaled antibiotics to treat ventilator-associated pneumonia?

BACKGROUND: The increasing prevalence of ventilator-associated pneumonia (VAP) due to either multidrug-resistant (MDR) organisms or infections with limited treatment options (i.e. susceptible to only aminoglycosides or colisitin) coupled with a dearth of new antimicrobials has led clinicians to pursue alternative management strategies including the use of inhaled antibiotics (IA). OBJECTIVES: To review the evidence surrounding the use of IA in the treatment of VAP with a focus on establishing a path whereby adjunctive IA could become a standard therapy for the treatment of specific VAP patient populations. SOURCES: A meta-analysis performed by the 2016 IDSA/ATS Hospital-acquired Pneumonia Guideline Committee; a PubMed and clinicaltrials.gov search for subsequent trials of IA for the treatment of VAP. CONTENT: Based on a meta-analysis of nine studies (RR 1.29; 95% CI 1.13-1.47), the 2016 IDSA/ATS Hospital-acquired Pneumonia Guideline Committee recommended that adjunctive IA be used to treat VAP due to Gram-negative bacilli that are susceptible to only aminoglycosides or polymyxins. Two subsequent randomized trials of adjunctive IA for the treatment of mechanically ventilated patients with pneumonia failed to demonstrate a benefit. Despite these results, an updated meta-analysis (n = 11) including these two recent trials suggests a benefit of adjunctive IA for the treatment of VAP due to MDR and difficult-to-treat infections (RR 1.2; 95% CI 1.05-1.57). IMPLICATIONS: Patients with VAP and limited intravenous antibiotic options are the individuals most likely to benefit from adjunctive IA and should be the focus of future investigative studies. Although vibrating mesh nebulizers predominate in pharmaceutical company-sponsored trials, these devices have not been directly compared with the traditional jet nebulizers in terms of efficacy or safety.

National profile of foot orthotic provision in the United Kingdom, part 2: podiatrist, orthotist and physiotherapy practices.

BACKGROUND: A national survey recently provided the first description of foot orthotic provision in the United Kingdom. This article aims to profile and compare the foot orthoses practice of podiatrists, orthotists and physiotherapists within the current provision. METHOD: Quantitative data were collected from podiatrists, orthotists and physiotherapists via an online questionnaire. The topics, questions and answers were developed through a series of pilot phases. The professions were targeted through electronic and printed materials advertising the survey. Data were captured over a 10 month period in 2016. Differences between professions were investigated using Chi squared and Fischer's exact tests, and regression analysis was used to predict the likelihood of each aspect of practice in each of the three professions. RESULTS: Responses from 357 podiatrists, 93 orthotists and 49 physiotherapists were included in the analysis. The results reveal statistically significant differences in employment and clinical arrangements, the clinical populations treated, and the nature and volume of foot orthoses caseload. CONCLUSION: Podiatrists, orthotists and physiotherapists provide foot orthoses to important clinical populations in both a prevention and treatment capacity. Their working context, scope of practice and mix of clinical caseload differs significantly, although there are areas of overlap. Addressing variations in practice could align this collective workforce to national allied health policy.

World association for the advancement of veterinary parasitology (WAAVP) guideline for testing the efficacy of ectoparasiticides for fish.

This guideline is intended to assist in the planning and execution of studies designed to assess the efficacy of ectoparasiticides for fish. It is the first ectoparasite-specific guideline to deal with studies set in the aquatic environment and therefore provides details for the maintenance of environmental standards for finfish. Information is included on a range of pre-clinical study designs as well as clinical studies in commercial/production sites, set within a regulatory framework. It provides information on the study animals, their welfare, husbandry and environmental requirements during the study. The most commonly pathogenic ectoparasites are presented with relevant points regarding life history, host challenge and numeric evaluation. Preparation and presentation of both topical and oral test treatments is provided, together with guidance on data collection and analysis. The guideline provides a quality standard or efficacy studies on finfish, which will assist researchers and regulatory authorities worldwide and contribute to the wider objective of harmonisation of procedures.

Multidose pharmacokinetics of orally administered florfenicol in the channel catfish (Ictalurus punctatus).

Plasma disposition of florfenicol in channel catfish was investigated after an oral multidose (10 mg/kg for 10 days) administration in freshwater at water temperatures ranging from 24.7 to 25.9 °C. Florfenicol concentrations in plasma were analyzed by means of liquid chromatography with MS/MS detection. After the administration of florfenicol, the mean terminal half-life (t(1/2)), maximum concentration at steady-state (Css (max)), time of Css (max) (T(max)), minimal concentration at steady-state (Css (min)), and Vc /F were 9.0 h, 9.72 µg/mL, 8 h, 2.53 µg/mL, and 0.653 L/kg, respectively. These results suggest that florfenicol administered orally at 10 mg/kg body weight for 10 days could be expected to control catfish bacterial pathogens inhibited in vitro by a minimal inhibitory concentration value of <2.5 µg/mL.

A risk-based approach to improve monitoring and performance of remote waste stabilisation ponds.

A cost-effective risk-based system was developed for assessing the performance and potential environmental impact of a large number of geographically dispersed pond systems, where cost and logistical issues prevent direct monitoring. In the process, a range of risk functions were calculated for each site to take into account pond performance, receiving environment, influent quality, surrounding land use and system size. Pond performance was estimated using traditional design equations, including Monte Carlo analysis to account for uncertainty in boundary conditions. The calculation of combined risk functions for all systems enabled the quantitative ranking of systems, which can be used to prioritise limited sampling resources.

Single intravenous and oral dose pharmacokinetics of florfenicol in the channel catfish (Ictalurus punctatus).

Plasma distribution and elimination of florfenicol in channel catfish were investigated after a single dose (10 mg/kg) of intravenous (i.v.) or oral administration in freshwater at a mean water temperature of 25.4 °C. Florfenicol concentrations in plasma were analyzed by means of liquid chromatography with MS/MS detection. After i.v. florfenicol injection, the terminal half-life (t(1/2)), volume of distribution at steady state (V(ss)), and central volume of distribution (V(c)) were 8.25 h, 0.9 and 0.381 L/kg, respectively. After oral administration of florfenicol, the terminal t(1/2), C(max), T(max), and oral bioavailability (F) were 9.11 h, 7.6 µg/mL, 9.2 h, and 1.09, respectively. There was a lag absorption time of 1.67 h in oral dosing. Results from these studies support that 10 mg florfenicol/kg body weight in channel catfish is an efficacious dosage following oral administration.

The effect of long-term wear of soft lenses of low and high oxygen transmissibility on the corneal epithelium.

PURPOSE: To measure the effects of long-term wear of soft lenses of low and high oxygen transmissibility (Dk) on basal epithelial appearance and epithelial thickness. METHODS: Sixty-three subjects were enrolled in a cross-sectional study. Seventeen high Dk lens wearers and 24 low Dk lens wearers who had worn lenses on an extended wear basis for more than 3 years (range: 3-22) were compared to a group of 22 age-matched subjects who had never worn contact lenses. Cell regularity and the intensity of light backscattered by the basal epithelium were assessed using confocal microscopy. Epithelial thickness was measured at the centre and at four peripheral locations using modified optical pachometry. RESULTS: Epithelial basal cells appeared less regular in low Dk lens wearers than high Dk wearers or controls (Mann-Whitney U-test, P=0.001). The intensity of backscattered light did not differ across groups (Kruskal-Wallis test, P=0.37). Low Dk wearers had the thinnest epithelium (46 (10) microm), followed by high Dk wearers (54 (14) microm) and controls (58 (9) microm; ANOVA, P

Identification of Porphyromonas levii isolated from clinical cases of bovine interdigital necrobacillosis by 16S rRNA sequencing.

Laboratories use pigmentation, antibiotic susceptibility, and biochemical tests to identify anaerobic organisms that play a role in bovine interdigital necrobacillosis (bovine foot rot). In this study, 16S rRNA gene sequencing was used to identify strains to the species level that were originally classified as Prevotella or Porphyromonas spp by conventional phenotype assessment methods. Of 264 qualified strains from ceftiofur clinical trials, 241 isolates were definitively identified by 16S rRNA sequencing as Porphyromonas levii. Similarly, of 275 qualified strains from tulathromycin clinical trials, 156 isolates were definitively identified by 16S rRNA sequencing as P. levii. The predominance of P. levii in this study supports the role of this organism as an associative agent of bovine foot rot and may have implications for routine laboratory diagnosis.

5-Azacytidine-treated human mesenchymal stem/progenitor cells derived from umbilical cord, cord blood and bone marrow do not generate cardiomyocytes in vitro at high frequencies.

BACKGROUND AND OBJECTIVES: Mesenchymal stem/progenitor cells (MSCs) are multipotent progenitors that differentiate into such lineages as bone, fat, cartilage and stromal cells that support haemopoiesis. Bone marrow MSCs can also contribute to cardiac repair, although the mechanism for this is unclear. Here, we examine the potential of MSCs from different sources to generate cardiomyocytes in vitro, as a means for predicting their therapeutic potential after myocardial infarction. MATERIALS AND METHODS: Mesenchymal stem/progenitor cells were isolated from the perivascular tissue and Wharton's jelly of the umbilical cord and from cord blood. Their immunophenotype and differentiation potential to generate osteoblasts, chondrocytes, adipocytes and cardiomyoxcytes in vitro was compared with those of bone marrow MSCs. RESULTS: Mesenchymal stem/progenitor cells isolated from umbilical cord and cord blood were phenotypically similar to bone marrow MSCs, the exception being in the expression of CD106, which was absent on umbilical cord MSCs, and CD146 that was highly expressed in cord blood MSCs. They have variable abilities to give rise to osteoblasts, chondrocytes and adipocytes, with bone marrow MSCs being the most robust. While a small proportion (approximately 0.07%) of bone marrow MSCs could generate cardiomyocyte-like cells in vitro, those from umbilical cord and cord blood did not express cardiac markers either spontaneously or after treatment with 5-azacytidine. CONCLUSION: Although MSCs may be useful for such clinical applications as bone or cartilage repair, the results presented here indicate that such cells do not generate cardiomyocytes frequently enough for cardiac repair. Their efficacy in heart repair is likely to be due to paracrine mechanisms.

Screening for autism in young children with developmental delay: an evaluation of the developmental behaviour checklist: early screen.

The ability to identify children who require specialist assessment for the possibility of autism at as early an age as possible has become a growing area of research. A number of measures have been developed as potential screening tools for autism. The reliability and validity of one of these measures for screening for autism in young children with developmental problems was evaluated. The parents of 207 children aged 20-51 months completed the Developmental Checklist-Early Screen (DBC-ES), prior to their child undergoing assessment. Good interrater agreement and internal consistency was found, along with significant correlations with a clinician completed measure of autism symptomatology. High sensitivity was found, with lower specificity for the originally proposed 17-item screening tool and a five-item version.

Temporal and spatial variation of physical, biological, and chemical parameters in a large waste stabilisation pond, and the implications for WSP modelling.

The spatial and temporal variation of physical, chemical, and biological parameters was determined, in summer and winter, at nine sites in a large (112 ha) waste stabilisation pond (WSP) at the Bolivar Wastewater Treatment Plant. Each site was extensively sampled over the course of one day, with the nine sites sampled over successive days at exactly the same times of day, progressing in the direction of bulk flow through the pond. Analyses of covariance were used to test the independent impact of site and climate on the way in which the mean values and stratification gradient of the physical, chemical, and biological parameters varied diurnally at each site. In both winter and summer studies there was a very strong correlation at all sites between changes in temperature, pH and dissolved oxygen (DO). Mean pond temperatures were higher in summer than winter, and thermal stratification was more common in summer. In summer, during the day at each site, concentrations of chlorophyll-a, DO, suspended solids and pH increased with higher solar radiation levels. This relationship was less evident in winter. There was no systematic depth or temporal variation identified in either the summer or winter study for the broad range of chemical parameters measured. Mean values for these parameters, and to a lesser extent their stratification gradients, increased by varying extents throughout the day at the different sites in both summer and winter, irrespective of changes in climate when the different sites were sampled. Sites nearer the inlet to the WSP recorded lower NH4N and higher NO2N and NO3N concentrations than the rest of the WSP. This was indicative of nitrification. Somewhat surprisingly, high DO concentrations were also recorded at these sites near the inlets. Computational fluid dynamics (CFD) modelling, incorporating the predominant wind conditions, offers a rationale for these observations. Recirculation was evident, which may increase the residence time for the slow growing autotrophic nitrifying bacteria and recirculate oxygen rich water around these sites - conditions which would enhance nitrification. Understanding the effect of these variations, overlaid by the influence of hydraulic and temporal scenarios, assists in developing a mechanistic understanding of pond operation.

Continuing medical education: a needs analysis of anaesthetists.

The continuing medical education (CME) needs of anaesthetists within Australia, New Zealand, Hong Kong, Malaysia and Singapore have been largely unknown. The aim of this study was to undertake a comprehensive survey of the attitude to CME, learning preferences, attitudes and abilities relating to self-paced material, literature and information searching, preferred content and preferred approach to CME of anaesthetists within these countries. A survey tool was developed and refined for ease of use by pilot-testing. The survey was mailed to 3,156 anaesthetists throughout Australia, New Zealand, Hong Kong, Malaysia and Singapore. Three options for data return were offered; postal reply, facsimile and a data entry web-page. There were 1,800 responses, which represented a response rate of 57%. The demographics of the respondents were similar to the overall demographics of Fellows of the Australian and New Zealand College of Anaesthetists. A large majority of respondents (92%) stated that their involvement in CME improved patient care. However, almost half the respondents reported that they have difficulty either in participating in current CME activities (31%) or implementing new knowledge into their workplace (14%). Anaesthetists within this region appear to be motivated by the need to make better decisions based on independent standards of practice. While Australia is a world leader in flexible education, it is still emerging as a discipline. Flexible education may be used to facilitate anaesthetists' participation in CME activities and in implementation of new knowledge in their workplace.

Behaviour and emotional problems in toddlers with pervasive developmental disorders and developmental delay: associations with parental mental health and family functioning.

BACKGROUND: Behavioural and emotional problems occur at a high rate in children and adolescents with intellectual disability, often from a young age. Some studies have indicated that children and adolescents with autism present with even higher rates. Less is known about the presentation, development and family impact of these difficulties in young children with autism. This study aimed to explore these issues in toddlers with pervasive developmental disorders (PDDs), those with delay without a PDD, and their families. METHODS: Participants were 123 children aged 20-51 months, referred to a developmental assessment clinic. Parents completed a checklist on child behavioural and emotional problems, and individual questionnaires on family functioning, their own mental health, and stress in relation to parenting their child. The child's language and cognitive skills, adaptive functioning and behaviour were assessed by standardized measures. Measures were repeated 1 year postdiagnosis. Behavioural and emotional problems in young children with a PDD were compared with those in children with developmental delay without a PDD, and their impact on parental outcomes explored over time. RESULTS: Initial and follow-up measures of child behaviour and emotional problems, parent mental health problems, parent stress and family functioning were significantly correlated, providing some evidence of stability over time. Child emotional and behavioural problems contributed significantly more to mother stress, parent mental health problems, and perceived family dysfunction than child diagnosis (PDD/non-PDD), delay or gender. Compared with mothers, all fathers reported significantly less stress in relation to parenting their child. CONCLUSION: Results highlighted the importance of addressing emotional and behavioural problems in very young children with autism and/or developmental delay. The need for early support and intervention for mothers, fathers and families in this context was also evidenced. As research has shown that behavioural and emotional problems persist into adolescence and young adulthood, understanding of these issues in very young children and their parents has important implications for intervention and long-term outcomes.

The identification of three novel MICA alleles by sequence-based typing.

During a study of MICA frequency in a healthy population and a cohort of patients suffering with inflammatory bowel disease, three DNA samples produced unusual reactivity patterns using polymerase chain reaction sequence-specific primers (PCR-SSP). These samples were subsequently characterized by sequence-based typing (SBT). Here, we report the sequence of these three novel MICA alleles.

Hyponatremic seizures after suprapubic catheter placement in 7-year-old child.

We report a case of hyponatremic seizures in a 7-year old boy with spina bifida following cystoscopy and suprapubic catheter placement. Immediate postoperative cystogram and pelvic computed tomogram (CT) after the development of seizures demonstrated a fluid collection from the suprapubic catheter site into the anterior abdominal wall. The subsequent reabsorption of free water from the fluid collection, with the contribution of postoperative hypotonic intravenous fluid administration and possible transient inappropriate antidiuretic hormone (ADH) secretion resulted in acute dilutional hyponatremia and consequent seizures. Strategies to prevent hyponatremia in children during urological procedures, with emphasis on the importance of reserving free water as the irrigation fluid are discussed.

Estimation of haemopoietic progenitor cells in peripheral blood by the Advia 120 and BD vantage flow cytometer: a direct comparison for the prediction of adequate collections.

Peripheral blood stem cells are increasingly used to ensure rapid haematological engraftment after myeloablative chemotherapy. After mobilization, progenitor cells in the blood can be enumerated to predict an adequate collection by leukapheresis. The Advia 120 automated counter has an immature cell channel measuring a parameter known as large undifferentiated cells (LUC's), which were quantified to assess their value in refining the timing of apheresis. Data were available from 102 apheresis sessions. Positive correlation was found for peripheral blood CD34+ cells and apheresis counts (r = 0.82, P < 0.0005) but not for total WCC (r = -0.15, P = 0.13) or LUC count (r = 0.12, P = 0.23). Our results indicate that the LUC population in peripheral blood has no relevance to the subsequent CD34 content of the apheresis product and CD34 cell enumeration by flow cytometry is advocated.

Changes in waste stabilisation pond performance resulting from the retrofit of activated sludge treatment upstream: part II--Management and operating issues.

Bolivar Wastewater Treatment Plant (WWTP) was originally commissioned with trickling filter secondary treatment, followed by waste stabilisation pond (WSP) treatment and marine discharge. In 1999, a dissolved air flotation/filtration (DAFF) plant was commissioned to treat a portion of the WSP effluent for horticultural reuse. In 2001, the trickling filters were replaced with activated sludge treatment. A shift in WSP ecology became evident soon after this time, characterised by a statistically significant reduction in algal counts in the pond effluent, and increased variability in algal counts and occasional population crashes in the ponds. While the photosynthetic capacity of the WSPs has been reduced, the concomitant reduction in organic loading has meant that the WSPs have not become overloaded. As a result of the improvement in water quality leaving the ponds, significant cost savings and improved product water quality have been realised in the subsequent DAFF treatment stage. A number of operating issues have arisen from the change, however, including the re-emergence of a midge fly nuisance at the site. Control of midge flies using chemical spraying has negated the cost savings realised in the DAFF treatment stage. While biomanipulation of the WSP may provide a less aggressive method of midge control, this case demonstrates the difficulty of predicting in advance all ramifications of a retrospective process change.

Changes in waste stabilisation pond performance resulting from the retrofit of activated sludge treatment upstream: part I--water quality issues.

This paper describes changes in effluent quality occurring before and after an upgrade to the Bolivar Wastewater Treatment Plant in South Australia. Trickling filters (TF) were replaced with an activated sludge (AS) plant, prior to tertiary treatment using waste stabilisation ponds (WSPs). The water quality in the WSPs following the upgrade was significantly improved. Reductions in total and soluble BOD, COD, TKN, suspended solids and organic nitrogen were recorded and the predominant form of inorganic nitrogen changed from NH(4)-N to NO(2)/NO(3)-N. The reduction in ammonium and potentially toxic free ammonia removed a control upon the growth of zooplankton, which may have contributed to decreases in algal biomass in the final ponds and consequently lower dissolved oxygen. Additionally, changes in inorganic nitrogen speciation contributed to a slightly elevated pH which reduced numbers of faecal coliforms in WSPs. The AS pretreated influent recorded significantly lower inorganic molar N:P ratio (10-4:1) compared to those fed with TF effluent (17-13:1). Algae within the WSPs may now be nitrogen limited, a condition which may favour the growth of nitrogen-fixing cyanobacteria. The decrease in algal biomass and in dissolved oxygen levels may enhance sedimentary denitrification, further driving the system towards nitrogen limitation.