Assessment of biological organ age using molecular pathology in pre-transplant kidney biopsies.

Organ shortage is a major challenge in kidney transplantation but the use of older donors, often with co-morbidities, is hampered by inconsistent outcomes. Methods of accurately stratifying marginal donor organs by clinical and histological assessment are lacking. To better understand organ variability, we profiled the transcriptomes of 271 kidneys from deceased donors at retrieval. Following correction for biopsy composition, we assessed molecular pathways that associated with delayed, and sub-optimal one-year graft function. Analysis of cortical biopsies identified an adaptive immune gene-rich module that significantly associated with increasing age and worse outcomes. Cellular deconvolution using human kidney reference single cell transcriptomes confirmed an increase in kidney-specific B and T cell signatures, as well as kidney macrophage, myofibroblast and fibroblast gene sets in this module. Surprisingly, innate immune pathway and neutrophil gene signature enrichment was associated with better outcomes. Thus, our work uncovers cellular molecular features of pathological organ ageing, identifiable at kidney retrieval, with translational potential.

Open Bedside Tracheostomy: Safe and Cost Saving but Underutilized Nationally.

OBJECTIVE: To evaluate open bedside tracheostomy (OBT) and compare it with open operating room (OR) tracheostomy and bedside percutaneous dilatational tracheostomy (PDT) in complications and cost. To determine the tracheostomy practice patterns of academic otolaryngology programs. STUDY DESIGN: Retrospective cohort study and cross-sectional study. SETTING: Public hospital and tertiary care hospital. METHODS: Otolaryngology program directors were surveyed to determine their institutions' tracheostomy practice patterns and the factors preventing the implementation of open bedside tracheostomies. A retrospective chart review was done of tracheostomies performed at our institutions from 2009 to 2019 for prolonged mechanical ventilation. Complications, length of intubation, comorbidities, body mass index, demographics, mortality rates, and decannulation rates were recorded. A cost analysis between OBT and PDT was conducted. RESULTS: Data from 802 patients were analyzed for 449 OBTs, 206 PDTs, and 147 open OR tracheostomies. Complication rates were low. PDTs were more likely to have perioperative tracheal bleeding (P = .028) and mucus plugging (P = .006). OBTs were performed on sicker patients with a higher Charlson Comorbidity Index than PDT and OR tracheostomies. The cost of OBT was less than that of PDT. The survey response rate of tracheostomy practice patterns was 46%. The otolaryngologists at the responding programs all conducted OR tracheostomies, while 52.7% did OBTs and 30.9% PDTs. CONCLUSION: OBT can be done safely in patients with multiple comorbidities and has a cost that can be less than PDT. Despite these benefits, only 50% of academic institutions routinely performed OBT.

Virtual Versus In-Person Head and Neck Physical Examination Training in Medical Students: A Pilot Assessment.

OBJECTIVE: To compare the effect of virtual and in-person head and neck physical examination training events on medical student confidence in performing examination maneuvers and seeking mentorship from otolaryngology faculty and residents. METHODS: Training events were held with first-year medical student volunteers in 2020 (in-person) and 2021 (virtual). Participants in both cohorts were given didactics on head and neck cancer, trained to perform a head and neck physical examination, and demonstrated their clinical skills to otolaryngology faculty and residents. Pre- and post-training surveys were utilized to assess the following outcomes: participant head and neck cancer knowledge, confidence in performing examination maneuvers, and confidence in seeking mentorship in otolaryngology. Differences in outcomes between training settings were assessed by comparing participant survey responses pre- and post- training. RESULTS: Both in-person and virtual training modalities improved participant confidence in performing the physical examination. There was no significant difference in the degree of improvement between training types. In-person training significantly increased participant confidence in seeking mentorship from otolaryngology faculty and residents (P = .003), while virtual training did not (P = .194). CONCLUSION: Virtual training modalities are feasible methods of teaching the head and neck physical examination. Instruction through a video conferencing platform has the potential to be incorporated into traditional in-person medical education in a permanent fashion. This pilot study can inform future studies directly comparing in-person and virtual physical examination training modalities.

Supraclavicular Artery Island Flap: Critical Appraisal and Comparison to Alternate Reconstruction.

OBJECTIVES/HYPOTHESIS: The supraclavicular artery island (SAI) flap may be a good option for selected head and neck reconstruction due to its reliability, ease of harvest, and favorable color match. The objective of this study was to examine the rates of complications for the SAI flap in head and neck oncologic reconstruction, with examination of risk factors and comparisons to alternative flaps often considered the gold-standard soft-tissue flaps for head and neck reconstruction: the pectoralis myocutaneous (PMC), radial forearm free flap (RFFF), and anterolateral thigh (ALT) flaps. STUDY DESIGN: Retrospective cohort study. METHODS: Consecutive SAI flaps were compared to PMC, RFFF, and ALT flaps (non-SAI flap group), all performed by the senior author from 2010 to 2018. The non-SAI flaps were included if an SAI flap could have been performed as an alternate flap. The groups were compared based on demographics, flap dimensions, site of reconstruction, operating time, total hospital stay, total hospital costs, and complications. RESULTS: One hundred seven SAI flaps and 194 non-SAI flaps were identified. SAI flaps were used less commonly than non-SAI flaps for mucosal defects (P < .001). The SAI flap dimensions were narrower but longer than non-SAI flaps (P < .001). SAI flaps had higher rates of total complications, partial flap necrosis, flap dehiscence at the recipient site, fistula, donor site dehiscence, and minor complications compared to non-SAI flaps (all P < .05). SAI flaps had higher rates of total complications, recipient site dehiscence, fistula, and minor complications in both the oral cavity and all mucosal sites compared to non-SAI flaps (all P < .05). SAI flaps for mucosal reconstruction were associated with higher rates of total complications (54% vs. 34%, P = .04), flap dehiscence at the recipient site (32% vs. 14%, P = .03), and major complications (21% vs. 5%, P = .02), compared to cutaneous reconstruction. Complications were equivalent between SAI flaps and non-SAI flaps for cutaneous reconstruction (all P > .05). Multivariate analysis showed that SAI flaps were associated with any postoperative complication (odds ratio [OR]: 3.47, 95% confidence interval [CI]: 1.85-6.54), partial flap necrosis (OR: 5.69, 95% CI: 1.83-17.7), flap dehiscence (OR: 5.36, 95% CI: 2.29-12.5), donor site complications (OR: 11.6, 95% CI: 3.27-41.0), and minor complications (OR: 5.17, 95% CI: 2.42-11.0). Within the SAI flap group, SAI flap length >24 cm was associated with postoperative complications on multivariate analysis (OR: 5.09, 95% CI: 1.02-25.5, P = .048). CONCLUSIONS: The SAI flap is best suited for cutaneous reconstruction of the face, neck, and parotid/temporal bone regions due to the favorable color match; the thin, pliable nature of the skin; ease of harvest; and equivalent complication rates compared to alternate soft-tissue flaps. However, the SAI flap is associated with more complications for oral cavity and mucosal site reconstruction when compared to RFFF and ALT flaps and should be used in selected cases that do not require complex folding. For all sites, flaps longer than 24 cm should be used with caution. LEVEL OF EVIDENCE: 3 Laryngoscope, 132:S1-S14, 2022.

Resolving cellular systems by ultra-sensitive and economical single-cell transcriptome filtering.

Single-cell transcriptomics suffer from sensitivity limits that restrict low abundance transcript identification, affects clustering and can hamper downstream analyses. Here, we describe Constellation sequencing (Constellation-Seq), a molecular transcriptome filter that delivers two orders of magnitude sensitivity gains by maximizing read utility while reducing the data sparsity and sequencing costs. The technique reliably measures changes in gene expression and was demonstrated by resolving rare dendritic cell populations from a peripheral blood mononuclear cell sample sample and exploring their biology with extreme resolution. The simple and powerful method is fully compatible with standard scRNA-Seq library preparation protocols and can be used for hypothesis testing, marker validation or investigating pathways.

Integrating human-centred design into the development of an intervention to improve the mental wellbeing of young women in the perinatal period: the Catalyst project.

BACKGROUND: Mental wellbeing during pregnancy and the year after birth is critical to a range of maternal and infant outcomes. Many mental health interventions fail to incorporate stakeholder perspectives. The Catalyst Project aimed to work with key stakeholders in Mozambique to develop interventions and delivery strategies which were in-line with existing evidence and the needs, goals, and priorities of those both directly and indirectly involved in its success. METHODS: A qualitative, human-centred design approach was utilised. Focus-group discussions, individual interviews, and observations with young women (aged 16-24 years), their families, community leaders, service providers and government were used to better understand the needs, priorities and challenges to mental wellbeing of young women. These findings were triangulated with the literature to determine priority challenges to be addressed by an intervention. Stakeholder workshops were held to identify potential solutions and co-develop an intervention and delivery strategy. RESULTS: The 65 participants comprised 23 young pregnant women or new mothers, 12 family members, 19 service providers and 11 staff from the Ministry of Health. Participants highlighted significant uncertainty related to living situations, financial status, education, social support, and limited knowledge of what to expect of the impact of pregnancy and parenting. Family and community support were identified as an important need among this group. The Mama Felíz (Happy Mama) programme was developed with stakeholders as a course to strengthen pregnancy, childbirth and child development knowledge, and build positive relationships, problem-solving and parenting skills. In addition, family sessions address wider cultural and gender issues which impact adolescent maternal wellbeing. CONCLUSIONS: We have developed an intervention to reduce the risk of poor maternal mental health and gives young mothers hope and skills to make a better life for them and their children by packaging information about the risk and protective factors for maternal mental disorders in a way that appeals to them, their families and service providers. By using human-centred design to understand the needs and priorities of young mothers and the health and community systems in which they live, the resulting intervention and delivery strategy is one that stakeholders view as appropriate and acceptable.

Detecting Wheat Powdery Mildew and Predicting Grain Yield Using Unmanned Aerial Photography.

High-resolution aerial imaging with an unmanned aerial vehicle (UAV) was used to quantify wheat powdery mildew and estimate grain yield. Aerial digital images were acquired at Feekes growth stage (GS) 10.5.4 from flight altitudes of 200, 300, and 400 m during the 2009-10 and 2010-11 seasons; and 50, 100, 200, and 300 m during the 2011-12, 2012-13, and 2013-14 seasons. The image parameter lgR was consistently correlated positively with wheat powdery mildew severity and negatively with wheat grain yield for all combinations of flight altitude and year. Fitting the data with random coefficient regression models showed that the exact relationship of lgR with disease severity and grain yield varied considerably from year to year and to a lesser extent with flight altitude within the same year. The present results raise an important question about the consistency of using remote imaging information to estimate disease severity and grain yield. Further research is needed to understand the nature of interyear variability in the relationship of remote imaging data with disease or grain yield. Only then can we determine how the remote imaging tool can be used in commercial agriculture.

Guidance on quantitative pest risk assessment.

This Guidance describes a two-phase approach for a fit-for-purpose method for the assessment of plant pest risk in the territory of the EU. Phase one consists of pest categorisation to determine whether the pest has the characteristics of a quarantine pest or those of a regulated non-quarantine pest for the area of the EU. Phase two consists of pest risk assessment, which may be requested by the risk managers following the pest categorisation results. This Guidance provides a template for pest categorisation and describes in detail the use of modelling and expert knowledge elicitation to conduct a pest risk assessment. The Guidance provides support and a framework for assessors to provide quantitative estimates, together with associated uncertainties, regarding the entry, establishment, spread and impact of plant pests in the EU. The Guidance allows the effectiveness of risk reducing options (RROs) to be quantitatively assessed as an integral part of the assessment framework. A list of RROs is provided. A two-tiered approach is proposed for the use of expert knowledge elicitation and modelling. Depending on data and resources available and the needs of risk managers, pest entry, establishment, spread and impact steps may be assessed directly, using weight of evidence and quantitative expert judgement (first tier), or they may be elaborated in substeps using quantitative models (second tier). An example of an application of the first tier approach is provided. Guidance is provided on how to derive models of appropriate complexity to conduct a second tier assessment. Each assessment is operationalised using Monte Carlo simulations that can compare scenarios for relevant factors, e.g. with or without RROs. This document provides guidance on how to compare scenarios to draw conclusions on the magnitude of pest risks and the effectiveness of RROs and on how to communicate assessment results.

Pest risk assessment of Spodoptera frugiperda for the European Union.

EFSA was asked for a partial risk assessment of Spodoptera frugiperda for the territory of the EU focussing on the main pathways for entry, factors affecting establishment, risk reduction options and pest management. As a polyphagous pest, five commodity pathways were examined in detail. Aggregating across these and other pathways, we estimate that tens of thousands to over a million individual larvae could enter the EU annually on host commodities. Instigating risk reduction options on sweetcorn, a principal host, reduces entry on that pathway 100-fold. However, sweetcorn imports are a small proportion of all S. frugiperda host imports, several of which are already regulated and further regulation is estimated to reduce the median number entering over all pathways by approximately 10%. Low temperatures limit the area for establishment but small areas of Spain, Italy and Greece can provide climatic conditions suitable for establishment. If infested imported commodities are distributed across the EU in proportion to consumer population, a few hundreds to a few thousands of individuals would reach NUTS 2 regions within which suitable conditions for establishment exist. Although S. frugiperda is a known migrant, entry directly into the EU from extant populations in sub-Saharan Africa is judged not feasible. However, if S. frugiperda were to establish in North Africa, in the range of thousands to over two million adults could seasonally migrate into the southern EU. Entry into suitable NUTS2 areas via migration will be greater than via commercial trade but is contingent on the establishment of S. frugiperda in North Africa. The likelihood of entry of the pest via natural dispersal could only be mitigated via control of the pest in Africa. If S. frugiperda were to arrive and become a pest of maize in the EU, Integrated Pest Management (IPM) or broad spectrum insecticides currently used against existing pests could be applied.

Pest risk assessment of Eotetranychus lewisi for the EU territory.

Following the 2014 EFSA's Panel on Plant Health scientific opinion on the pest categorisation of the spider mite Eotetranychus lewisi, the European Commission requested the Panel to perform a pest risk assessment and evaluate the risk reduction options. A stochastic model was used to assess entry, establishment and spread and related uncertainties. In the EU, E. lewisi has only been reported to occur in Portugal (Madeira). Entry pathways assessed were strawberry plants for planting from the USA, poinsettia and raspberry plants for planting, and orange and lemon fruits from third countries. Entry is most likely via poinsettia. Under current EU phytosanitary requirements, there is around a one in ten chance that E. lewisi will establish outdoors over the next 10 years. Although unlikely, establishment would most likely occur in southern Europe where environmental conditions, temperature and host density, are most suitable. If E. lewisi did establish, pest spread is expected to be mainly human assisted, most likely the mite being transported long distances on plants for planting. Nevertheless, while remaining a regulated pest, spread would be slow and most likely confined to one NUTS 2 area after 10 years. Under a scenario with enhanced measures (pest free place of production) at origin, the Panel's assessment indicate that it is extremely unlikely that E. lewisi would establish within 10 years hence spread is also extremely unlikely. The absence of trade of host plants from Madeira to other parts of the EU could explain why E. lewisi has not spread to other EU Member States. E. lewisi is reported as reducing yield and quality of peaches and poinsettia and is regarded as a growing concern for strawberry and raspberry growers in the Americas. The Panel concludes that should E. lewisi be introduced in the EU similar impacts could be expected.

Pest risk assessment of Atropellis spp. for the EU territory.

Following a request from the European Commission, the EFSA Plant Health (PLH) Panel performed a risk assessment for Atropellis spp. in the EU focusing on the risk of entry, the host range and the potential impacts. Atropellis is a fungal pathogen of several Pinus spp. in North America. The pathogen has not been reported from Europe and is a quarantine pest regulated in Annex IIAI of Council Directive 2000/29/EC on plants (other than fruit and seeds), isolated bark and wood of Pinus. The main pathways of entry considered were Pinus plants, wood and isolated bark. Given the ban of importing Pinus plants from outside Europe into the EU and the lack of information on EU imports of isolated Pinus bark, only the wood pathway was assessed quantitatively. The conclusion of the assessment of entry for scenario A0 (current regulatory situation) is that the risk of entry of Atropellis spp. is close to zero. This conclusion is expected to apply also in the case of removing the specific Atropellis regulations, because of the remaining generic Pinus requirements, as well as in a scenario with additional risk reduction options. The uncertainty associated with this assessment is relatively limited, given that all the quartiles of the estimated distribution of the number of potential founder populations are close to zero. For the North American Pinus spp. known to be susceptible and widely planted in the EU (mainly P. contorta and P. strobus), the damage observed in North America (loss of wood quality, stem deformations, mortality in young stands, environmental consequences) is expected in the EU to a similar (or higher) degree, should the pathogen be introduced. Similar impacts are expected on the European Pinus spp. known to be host of Atropellis spp. These include widespread and locally abundant species such as P. nigra, P. sylvestris and P. pinaster. There are, however, large uncertainties associated with this impact assessment due to the unknown susceptibility of several other Pinus spp. present in Europe. There is a need for research on the susceptibility to Atropellis spp. of those European Pinus spp.

Pest risk assessment of Radopholus similis for the EU territory.

The Panel on Plant Health performed a pest risk assessment on Radopholus similis, the burrowing nematode for the EU. The quantitative assessment focused on entry, establishment, spread and impact on tropical and subtropical ornamental host plants, the main pathways for entry of R. similis into the EU. Infested consignments are expected to enter the risk assessment area on ornamentals under all scenarios. For citrus, which is a closed pathway for entry, outdoor establishment was assessed. Establishment may only take place after successful transfer from ornamental plants to citrus production systems. This event is called 'shift' in this assessment, to indicate that this is an unusual transfer. It has been estimated that establishment of this nematode in the open field in the EU citrus production areas under current temperatures is possible in most parts of the citrus production area in the EU. Temperature conditions will prevent the nematode from establishing only in the northernmost citrus areas and at higher altitudes in the south. Host plants for planting originating from infested places of production (greenhouses) within the risk assessment area are considered the main pathway for spread within the risk assessment area. Under current climatic conditions, the population of R. similis is not expected to reach damaging population levels in the open field. In case of increased temperatures due to global warming, the nematode population may reach damaging levels in very few places outdoors. Currently, main impact is considered for ornamental greenhouse production in the risk assessment area. Impact will be either caused by direct plant growth reductions or loss due to phytosanitary measures applied on regulated plants. Despite the fact that R. similis is globally considered as one of the most destructive plant parasitic nematodes, the impact in the risk assessment area is considered low.

Pest risk assessment of Diaporthe vaccinii for the EU territory.

As requested by the European Commission, the EFSA Panel on Plant Health (PLH) Panel assessed the risk of Diaporthe vaccinii in the EU, focusing on entry, establishment, spread and impacts on cultivated and wild Vaccinium species, the principal hosts being American and European cranberry and blueberry. Several outbreaks occurred in the EU since 1956, but most were eradicated except in Latvia. The Panel considered entry via fruits and plants for planting. The risk of establishment from discarded infected berries is much lower than from infected plants for planting, of which, potted plants and cuttings pose the greatest risk, while plug plants, derived from tissue culture and grown in pest free structures, pose a low risk. Nine per cent of the EU is highly suitable for establishment of the pathogen, mostly in the SE and NE. Following establishment, the pathogen could spread naturally over short range, and by human assistance over long range. Calculations with an integrated model for entry, establishment and spread, indicate that with current regulations, over a period of 5 years, a few hundred cultivated Vaccinium plants and several thousand Vaccinium plants in natural ecosystems would contract the disease. The associated loss of commercial production is small, less than one tonne of berries per year. On natural vegetation, the median impact after 5 years was estimated to be negligible affecting a negligible proportion of the natural Vaccinium population (2 × 10-8). However, the uncertainty of this estimate was high, due to uncertainty about the rate of spread; in a worst-case scenario (99th percentile), almost 1% of plants in natural areas would become infected. Complete deregulation (scenario A1) was predicted to increase the impact substantially, especially in natural areas, while additional measures (scenario A2) would effectively eliminate the entry of infected plants for planting, further reducing the impacts below the current situation.

Declining health insurance access among US Hispanic workers: not all jobs are created equal.

INTRODUCTION: Approximately 18% of the U.S. population are uninsured, a proportion that may continue to rise, particularly among Hispanics, as the cost of medical care increases faster than the growth in wages. METHODS: Health insurance trends were analyzed by race-ethnic category, and among Hispanic workers by occupation type and industrial sector, using data on employed respondents > or =18 years from 1997 to 2007 National Health Interview Survey (NHIS) (mean annual n = 17,392, representing 123 million US workers on average over this 11 year period). RESULTS: From 1997 to 2007, the relative decline in health insurance coverage for US workers was greatest among Hispanics (7.0%). Hispanic workers in the Construction and Services industries had the greatest overall decline in coverage (24.9% and 14.7%), as well as Hispanic blue collar workers (14.0%). CONCLUSION: Hispanic workers in general, and those employed in blue collar, construction, and services sectors in particular, are at greater risk for poor access to health care due to a lack of health insurance coverage.

Massively parallel production of lipid microstructures.

In this paper we describe a simple and inexpensive microfluidic system for the production of lipid tubules and vesicles. The system incorporates a central microporous membrane for interfacing lipid films with aqueous flows. Hydrodynamic drag was used for the parallel elongation of high axial ratio lipid tubules with uniform 1.5 +/- 0.5 microm diameters. Alternatively, electrokinetic operation was used for the rapid and continuous production of vast numbers of lipid vesicles with diameters ranging from 1 to 3 microm.

Council tax valuation bands, socio-economic status and health outcome: a cross-sectional analysis from the Caerphilly Health and Social Needs Study.

UNLABELLED: Council tax valuation bands (CTVBs) are a categorisation of household property value in Great Britain. The aim of the study was to assess the CTVB as a measure of socio-economic status by comparing the strength of the associations between selected health and lifestyle outcomes and CTVBs with two measures of socio-economic status: the National Statistics Socio-Economic Classification (NS-SEC) and the 2001 UK census-based Townsend deprivation index. METHODS: Cross-sectional analysis of data on 12,092 respondents (adjusted response 62.7%) to the Caerphilly Health and Social Needs Study, a postal questionnaire survey undertaken in Caerphilly county borough, south-east Wales, UK. The CTVB was assigned to each individual by matching the sampling frame to the local authority council tax register. Crude and age-gender adjusted odds ratios for each category of CTVB, NS-SEC and fifth of the ward distribution of Townsend scores were estimated for smoking, poor diet, obesity, and limiting long-term illness using logistic regression. Mean mental (MCS) and physical (PCS) component summary scores of the Short-Form SF-36 health status questionnaire were estimated in general linear models. RESULTS: There were significant trends in odds ratios across the CTVB categories for all outcomes, most marked for smoking and mental and physical health status. The adjusted odds ratio for being a smoker in the lowest versus highest CTVB category was 3.80 (95% CI: 3.06, 4.71), compared to 3.00 (95% CI: 2.30, 3.90) for the NS-SEC 'never worked and long-term unemployed' versus 'higher managerial and professional' categories, and 1.61 (95% CI: 1.42, 1.83) for the most deprived versus the least deprived Townsend fifth. The difference in adjusted mean MCS scores was 5.9 points on the scale for CTVB, 9.2 for NS-SEC and 3.2 for the Townsend score. The values for the adjusted mean PCS scores were 6.3 points for CTVB, 11.3 for NS-SEC, and 2.5 for the Townsend score. CONCLUSION: CTVBs assigned to individuals were strongly associated with the health and lifestyle outcomes modelled in this study. CTVBs are readily available for all residential properties and deserve further consideration as a proxy for socio-economic status in epidemiological studies in Great Britain.