Prenatal methadone exposure and child developmental outcomes in 2-year-old children.

AIM: To examine the developmental outcomes of children born to opioid-dependent females enrolled in methadone maintenance and identify pre- and postnatal factors that place these children at developmental risk. METHOD: Ninety-nine methadone-maintained females and their 100 infants (42 females, 58 males, mean gestational age 38.8wks) were recruited during pregnancy/at birth and studied to age 2 years alongside a regionally representative comparison group of 108 non-methadone-maintained females and their 110 infants (62 females, 48 males, mean gestational age 39.2wks). Information about perinatal exposure was collected from medical records, maternal urine and infant meconium toxicological analysis, maternal interviews (at birth and at 18mo), and a home visit (at 18mo). At age 2 years, child neuromotor function, cognition, language, and emotional/behavioral dysregulation were assessed. RESULTS: Opioid-exposed children achieved lower motor, cognitive, and language scores and had poorer self, emotional, eating/feeding, and sensory processing regulation than unexposed children. After adjustment for maternal education and other substance use in pregnancy, between-group differences in child motor, cognitive, and overall dysregulation remained. Postnatal parental and family factors explained a further 40% to 52% of between-group differences in child outcomes. INTERPRETATION: These children and families are extremely high-risk and need antenatal and postnatal support. Children exposed to opioids during pregnancy have pervasive developmental difficulties by age 2 years. These challenges are largely explained by adverse pregnancy and socio-environmental exposures, emphasizing the importance of specialist prenatal care and postnatal intervention support. What this paper adds Children born to opioid-dependent females are at high risk of pervasive developmental problems. These problems span a range of functional domains, including motor, cognitive, language, and behavioral/emotional dysregulation. Contributing factors include other adverse pregnancy exposures, postnatal environmental factors, and the direct effects of prenatal opioid exposure.

Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding.

Pollen metabarcoding is emerging as a powerful tool for ecological research and offers unprecedented scale in citizen science projects for environmental monitoring via honey bees. Biases in metabarcoding can be introduced at any stage of sample processing and preservation is at the forefront of the pipeline. While in metabarcoding studies pollen has been preserved at - 20 °C (FRZ), this is not the best method for citizen scientists. Herein, we compared this method with ethanol (EtOH), silica gel (SG) and room temperature (RT) for preservation of pollen collected from hives in Austria and Denmark. After ~ 4 months of storage, DNAs were extracted with a food kit, and their quality and concentration measured. Most DNA extracts exhibited 260/280 absorbance ratios close to the optimal 1.8, with RT samples from Austria performing slightly worse than FRZ and SG samples (P < 0.027). Statistical differences were also detected for DNA concentration, with EtOH samples producing lower yields than RT and FRZ samples in both countries and SG in Austria (P < 0.042). Yet, qualitative and quantitative assessments of floral composition obtained using high-throughput sequencing with the ITS2 barcode gave non-significant effects of preservation methods on richness, relative abundance and Shannon diversity, in both countries. While freezing and ethanol are commonly employed for archiving tissue for molecular applications, desiccation is cheaper and easier to use regarding both storage and transportation. Since SG is less dependent on ambient humidity and less prone to contamination than RT, we recommend SG for preserving pollen for metabarcoding. SG is straightforward for laymen to use and hence robust for widespread application in citizen science studies.

Evaluation of the ability of three commercially available dosimeters to detect systematic delivery errors in step-and-shoot IMRT plans.

BACKGROUND: There is limited data on error detectability for step-and-shoot intensity modulated radiotherapy (sIMRT) plans, despite significant work on dynamic methods. However, sIMRT treatments have an ongoing role in clinical practice. This study aimed to evaluate variations in the sensitivity of three patient-specific quality assurance (QA) devices to systematic delivery errors in sIMRT plans. MATERIALS AND METHODS: Four clinical sIMRT plans (prostate and head and neck) were edited to introduce errors in: Multi-Leaf Collimator (MLC) position (increasing field size, leaf pairs offset (1-3 mm) in opposite directions; and field shift, all leaves offset (1-3 mm) in one direction); collimator rotation (1-3 degrees) and gantry rotation (0.5-2 degrees). The total dose for each plan was measured using an ArcCHECK diode array. Each field, excluding those with gantry offsets, was also measured using an Electronic Portal Imager and a MatriXX Evolution 2D ionisation chamber array. 132 plans (858 fields) were delivered, producing 572 measured dose distributions. Measured doses were compared to calculated doses for the no-error plan using Gamma analysis with 3%/3 mm, 3%/2 mm, and 2%/2 mm criteria (1716 analyses). RESULTS: Generally, pass rates decreased with increasing errors and/or stricter gamma criteria. Pass rate variations with detector and plan type were also observed. For a 3%/3 mm gamma criteria, none of the devices could reliably detect 1 mm MLC position errors or 1 degree collimator rotation errors. CONCLUSIONS: This work has highlighted the need to adapt QA based on treatment plan type and the need for detector specific assessment criteria to detect clinically significant errors.

An international survey on the clinical use of rigid and deformable image registration in radiotherapy.

OBJECTIVES: Rigid image registration (RIR) and deformable image registration (DIR) are widely used in radiotherapy. This project aims to capture current international approaches to image registration. METHODS: A survey was designed to identify variations in use, resources, implementation, and decision-making criteria for clinical image registration. This was distributed to radiotherapy centers internationally in 2018. RESULTS: There were 57 responses internationally, from the Americas (46%), Australia/New Zealand (32%), Europe (12%), and Asia (10%). Rigid image registration and DIR were used clinically for computed tomography (CT)-CT registration (96% and 51%, respectively), followed by CT-PET (81% and 47%), CT-CBCT (84% and 19%), CT-MR (93% and 19%), MR-MR (49% and 5%), and CT-US (9% and 0%). Respondent centers performed DIR using dedicated software (75%) and treatment planning systems (29%), with 84% having some form of DIR software. Centers have clinically implemented DIR for atlas-based segmentation (47%), multi-modality treatment planning (65%), and dose deformation (63%). The clinical use of DIR for multi-modality treatment planning and accounting for retreatments was considered to have the highest benefit-to-risk ratio (69% and 67%, respectively). CONCLUSIONS: This survey data provides useful insights on where, when, and how image registration has been implemented in radiotherapy centers around the world. DIR is mainly in clinical use for CT-CT (51%) and CT-PET (47%) for the head and neck (43-57% over all use cases) region. The highest benefit-risk ratio for clinical use of DIR was for multi-modality treatment planning and accounting for retreatments, which also had higher clinical use than for adaptive radiotherapy and atlas-based segmentation.

Reassessing Southern Ocean Air-Sea CO2 Flux Estimates With the Addition of Biogeochemical Float Observations.

New estimates of pCO2 from profiling floats deployed by the Southern Ocean Carbon and Climate Observations and Modeling (SOCCOM) project have demonstrated the importance of wintertime outgassing south of the Polar Front, challenging the accepted magnitude of Southern Ocean carbon uptake (Gray et al., 2018, https://doi:10.1029/2018GL078013). Here, we put 3.5 years of SOCCOM observations into broader context with the global surface carbon dioxide database (Surface Ocean CO2 Atlas, SOCAT) by using the two interpolation methods currently used to assess the ocean models in the Global Carbon Budget (Le Quéré et al., 2018, https://doi:10.5194/essd-10-2141-2018) to create a ship-only, a float-weighted, and a combined estimate of Southern Ocean carbon fluxes (<35°S). In our ship-only estimate, we calculate a mean uptake of -1.14 ± 0.19 Pg C/yr for 2015-2017, consistent with prior studies. The float-weighted estimate yields a significantly lower Southern Ocean uptake of -0.35 ± 0.19 Pg C/yr. Subsampling of high-resolution ocean biogeochemical process models indicates that some of the differences between float and ship-only estimates of the Southern Ocean carbon flux can be explained by spatial and temporal sampling differences. The combined ship and float estimate minimizes the root-mean-square pCO2 difference between the mapped product and both data sets, giving a new Southern Ocean uptake of -0.75 ± 0.22 Pg C/yr, though with uncertainties that overlap the ship-only estimate. An atmospheric inversion reveals that a shift of this magnitude in the contemporary Southern Ocean carbon flux must be compensated for by ocean or land sinks within the Southern Hemisphere.

The Four-Dimensional Carbon Cycle of the Southern Ocean.

The Southern Ocean plays a fundamental role in the global carbon cycle, dominating the oceanic uptake of heat and carbon added by anthropogenic activities and modulating atmospheric carbon concentrations in past, present, and future climates. However, the remote and extreme conditions found there make the Southern Ocean perpetually one of the most difficult places on the planet to observe and to model, resulting in significant and persistent uncertainties in our knowledge of the oceanic carbon cycle there. The flow of carbon in the Southern Ocean is traditionally understood using a zonal mean framework, in which the meridional overturning circulation drives the latitudinal variability observed in both air-sea flux and interior ocean carbon concentration. However, recent advances, based largely on expanded observation and modeling capabilities in the region, reveal the importance of processes acting at smaller scales, including basin-scale zonal asymmetries in mixed-layer depth, mesoscale eddies, and high-frequency atmospheric variability. Assessing the current state of knowledge and remaining gaps emphasizes the need to move beyond the zonal mean picture and embrace a four-dimensional understanding of the carbon cycle in the Southern Ocean.

High Rates of Honey Bee Colony Losses and Regional Variability in Ethiopia Based on the Standardised COLOSS 2023 Survey.

The COLOSS research association has been assessing honey bee colony losses, associated risk factors and management, focusing on Western countries but with a progressive international expansion. Here, we report the first survey on the loss rates of colonies in 2022/2023 in Ethiopia using COLOSS monitoring survey tools. A face-to-face interview questionnaire survey was conducted on 64 beekeepers selected from Oromia and Tigray regions. This covered 1713 honey bee colonies distributed in 68 apiaries. The percentages of colonies lost were significantly different between Oromia (24.1%) and Tigray (66.4%) regions. Colony losses were attributed as unsolvable queen problems (8% in Oromia; 10% in Tigray), natural disaster (32%; 82%), and empty hives or dead colonies (60%; 8%). The loss rate was significantly affected by queen replacement (p < 0.0001), use of natural comb (p < 0.0001), feed supplementation (p < 0.0001), region (p < 0.0001), varroa treatment (p < 0.0001), colony splitting (p < 0.01), and merging (p < 0.01). Beekeepers in Oromia managed more colonies and implemented improved practices compared to those in Tigray. However, all beekeepers in Oromia detected at least some bees with signs of deformed wing virus, compared to 76% of beekeepers in Tigray. In conclusion, the colony loss rate was significantly different between Oromia and Tigray regions due to differences in natural disasters, management, environment and health factors.

The assessment of the clinical impact of using a single set of radiotherapy planning data for two kilovoltage therapy units.

Kilovoltage therapy units are used for superficial radiotherapy treatment delivery. Peer reviewed studies for MV linear accelerators describe tolerances to dosimetrically match multiple linear accelerators enabling patient treatment on any matched machine. There is an absence of literature on using a single planning data set for multiple kilovoltage units which have limited ability for beam adjustment. This study reviewed kilovoltage dosimetry and treatment planning scenarios to evaluate the feasibility of using ACPSEM annual QA tolerances to determine whether two units (of the same make and model) were dosimetrically matched. The dosimetric characteristics, such as measured half value layer (HVL), percentage depth dose (PDD), applicator factor and output variation with stand-off distance for each kV unit were compared to assess the agreement. Independent planning data based on the measured HVL for each beam energy from each kV unit was prepared. Monitor unit (MU) calculations were performed using both sets of planning data for approximately 200 clinical scenarios and compared with an overall agreement between units of < 2%. Additionally, a dosimetry measurement comparison was completed at each site for a subset of nine scenarios. All machine characterisation measurements were within the ACPSEM Annual QA tolerances, and dosimetric testing was within 2.5%. This work demonstrates that using a single set of planning data for two kilovoltage units is feasible, resulting in a clinical impact within published uncertainty.

Seasonal Colony Loss Rates and Honey Bee Management in the Kingdom of Saudi Arabia: Results of a Survey of Beekeepers.

There is high demand for honey in Saudi Arabia, honey bees make a valuable contribution to agriculture and the economy, and therefore it is important to know levels of colony loss and potential reasons for losses. While there is much research into honey bee colony losses worldwide, little is known about colony losses in Saudi Arabia, management practices or beekeeping experience there. The aims of this work were to address this knowledge gap. Results of a survey of beekeepers in southwest Saudi Arabia conducted in summer 2018 are presented, including colony losses in five different seasons. Data collection involved face-to-face interviews, supplemented by an online survey, using a purpose-designed questionnaire. Responses were obtained from 109 beekeepers, all male, managing 135 to 1700 colonies, with 2 to 45 years of beekeeping experience. Most (73.1%) respondents mainly kept local hybrid bees, while 25.9% mainly kept Apis mellifera jemenitica. Honey yields per colony varied much more between beekeepers than between bee races. A high proportion (83.5%) of beekeepers reported losing colonies over the period studied. The reported colony loss rate was significantly higher in summer than in other seasons, but still low. The overall proportion of colonies lost was 11.4% in summer 2017 and was lowest in spring 2018 (6.6%). The main reported causes of loss were Varroa destructor and disease. Most beekeepers (88.0%) treated against the Varroa mite, although only one method was reported, tau-fluvalinate as Apistan strips, and only 41.7% used a screened bottom board. The results establish a benchmark for future beekeeper surveys in Saudi Arabia and other environmentally similar countries where colony losses are of interest in all seasons of the year. Informing and supporting Saudi beekeepers concerning Varroa monitoring and treatment and optimal hive management could result in fewer losses, higher honey yields, potential to market organic honey and a greater share of the domestic honey market.

Honey Bee Colony Losses in Mexico's Semi-Arid High Plateau for the Winters 2016-2017 to 2021-2022.

The objective of this study was to estimate the loss rates of honey bee (Apis mellifera) colonies in six consecutive winters (2016-2017 to 2021-2022) in five states of the semi-arid high plateau region of Mexico, as well as study the factors associated with these losses. The survey included data from 544 beekeepers and 75,341 colonies. The colony loss rate differs significantly (p < 0.0001) between the winters analyzed and fluctuates between 14.9% and 30%, with an average of 22%. Colony losses were unaffected (p > 0.05) by migratory beekeeping practice and operation size, but losses were significantly affected by Varroa monitoring and control (p ≤ 0.001). The types of loss differ among the winters analyzed. In the winters of 2016-2017 to 2018-2019, beekeepers attributed higher loss rates to unresolved problems related to the queen (e.g., a queenless colony, poor laying), but in the winters of 2019-2020 to 2021-2022, the highest loss rate was the result of problems such as diseases, poisoning, and absconding (leading to dead colonies or empty hives). The results reveal high loss rates in the region studied, as reported by beekeepers from other countries. It is suggested that strategies should be implemented to improve the quality of queens, the control of varroasis and other diseases, and the degree of Africanization.

A Simple Whole-Plasmid PCR Method to Construct High-Diversity Synthetic Phage Display Libraries.

Phage display technology utilises peptide and antibody libraries with very high diversities to select ligands with specific binding properties. The production of such libraries can be labour intensive and technically challenging and whilst there are commercial sources of libraries, the exploitation of the resulting binders is constrained by ownership of the libraries. Here, a peptide library of ~ 1 × 109 variants for display on gene VIII was produced alongside three VHH antibody libraries with similar diversity, where 12mer, 16mer or 21mer CDR3s were introduced into the highly stable cAbBCII10 scaffold displayed on gene III. The cloning strategy used a simple whole-plasmid PCR method and type IIS restriction enzyme assembly that facilitate the seamless insertion of diversity into any suitable phage coat protein or antibody scaffold. This method reproducibly produced 1 × 109 variants from just 10 transformations and the four libraries had relatively low bias with 82 to 86% of all sequences present as single copies. The functionality of both peptide and antibody libraries were demonstrated by selection of ligands with specific binding properties by biopanning. The peptide library was used to epitope map a monoclonal antibody. The VHH libraries were pooled and used to select an antibody to recombinant human collagen type 1.

CSI Pollen: Diversity of Honey Bee Collected Pollen Studied by Citizen Scientists.

A diverse supply of pollen is an important factor for honey bee health, but information about the pollen diversity available to colonies at the landscape scale is largely missing. In this COLOSS study, beekeeper citizen scientists sampled and analyzed the diversity of pollen collected by honey bee colonies. As a simple measure of diversity, beekeepers determined the number of colors found in pollen samples that were collected in a coordinated and standardized way. Altogether, 750 beekeepers from 28 different regions from 24 countries participated in the two-year study and collected and analyzed almost 18,000 pollen samples. Pollen samples contained approximately six different colors in total throughout the sampling period, of which four colors were abundant. We ran generalized linear mixed models to test for possible effects of diverse factors such as collection, i.e., whether a minimum amount of pollen was collected or not, and habitat type on the number of colors found in pollen samples. To identify habitat effects on pollen diversity, beekeepers' descriptions of the surrounding landscape and CORINE land cover classes were investigated in two different models, which both showed that both the total number and the rare number of colors in pollen samples were positively affected by 'urban' habitats or 'artificial surfaces', respectively. This citizen science study underlines the importance of the habitat for pollen diversity for bees and suggests higher diversity in urban areas.

Deforming to Best Practice: Key considerations for deformable image registration in radiotherapy.

Image registration is a process that underlies many new techniques in radiation oncology - from multimodal imaging and contour propagation in treatment planning to dose accumulation throughout treatment. Deformable image registration (DIR) is a subset of image registration subject to high levels of complexity in process and validation. A need for local guidance to assist in high-quality utilisation and best practice was identified within the Australian community, leading to collaborative activity and workshops. This report communicates the current limitations and best practice advice from early adopters to help guide those implementing DIR in the clinic at this early stage. They are based on the state of image registration applications in radiotherapy in Australia and New Zealand (ANZ), and consensus discussions made at the 'Deforming to Best Practice' workshops in 2018. The current status of clinical application use cases is presented, including multimodal imaging, automatic segmentation, adaptive radiotherapy, retreatment, dose accumulation and response assessment, along with uptake, accuracy and limitations. Key areas of concern and preliminary suggestions for commissioning, quality assurance, education and training, and the use of automation are also reported. Many questions remain, and the radiotherapy community will benefit from continued research in this area. However, DIR is available to clinics and this report is intended to aid departments using or about to use DIR tools now.

Evaluating evidence-based practice within critical care.

BACKGROUND AND AIMS: Between 2002-2005 the Trust undertook an action research project to evaluate a corporate practice development strategy. During this period clinicians became practitioner-researchers utilising a variety of methods to evaluate the influence of practice development. One aspect of this focused upon evaluation of evidence based guidelines. This article concentrates upon this process and the learning from this within critical care. METHOD: Within critical care it was recognised that the standard of guidelines and protocols varied in terms of the amount of evidence used to underpin decision making. A group was set up to evaluate and appraise these using a structured format such as the Appraisal of Guidelines for Research and Evaluation (AGREE) Instrument. FINDINGS: The initial evaluation (cycle 1) highlighted learning associated with the process of using the instrument within critical care, as well as where the quality of the guidelines could be improved. The second cycle of evaluation demonstrated that implementation of the action plans as a consequence of cycle 1 resulted in an improvement in the quality of the guidelines. It also resulted in streamlining the process of undertaking guideline appraisal across a Trust. DISCUSSION AND CONCLUSIONS: Action resulting from analysis of the findings of cycle 1 led to a cultural change in which the structure of a tool such as the AGREE instrument could be beneficial in the development of future guidelines. This has been sustained both within critical care and Trust wide with various initiatives such as the establishment of critical care multidisciplinary guideline development groups and a Trust wide electronic library management system.

No place like home: A case study of a patient discharged from an Intensive Care Unit for end of life care at home.

INTRODUCTION: Both in the UK and internationally, discharge from an intensive care unit to home for end of life care is a rare and challenging occurrence. These challenges include clinicians' ability to identify appropriate patients in whom it is possible to communicate with about their wishes and preferences, the critical nature of their condition and the interface between hospital and community services. METHOD: We present a case report of a patient who had been admitted to hospital with a myocardial infarction and subsequently suffered a cardiac arrest, from which he was successfully resuscitated. Subsequently, he suffered multi-organ failure, but despite treatments, the ceiling of care was reached. With a poor prognosis, medical and nursing staff engaged in advance care planning to determine his wishes and preferences at the end of life and to facilitate his discharge from the intensive care unit to his home. CONCLUSION: This case study has highlighted that through good communication amongst patients, families and professionals and collaborative working across boundaries and organisations, appropriate patients in the critical care setting can have a real choice regarding where they wish to be cared for and die at the end of their life.

Author Correction: Spiraling pathways of global deep waters to the surface of the Southern Ocean.

The original version of this Article contained errors in Fig. 6. In panel a, the grey highlights obscured the curves for CESM, CM2.6 and SOSE, and the labels indicating SWIR, KP, MR, PAR, and DP were inadvertently omitted. These have now been corrected in both the PDF and HTML versions of the Article.

Quantifying tumour-infiltrating lymphocyte subsets: a practical immuno-histochemical method.

BACKGROUND: Efficient histological quantification of tumour-infiltrating T and B lymphocyte (TIL) subsets in archival tissues would greatly facilitate investigations of the role of TIL in human cancer biology. We sought to develop such a method. METHODS: Ten x40 digital images of 4 micro sections of 16 ductal invasive breast carcinomas immunostained for CD3, CD4, CD8, and CD20 were acquired (a total of 640 images). The number of pixels in each image matching a partition of Lab colour space corresponding to immunostained cells were counted using the 'Color range' and 'Histogram' tools in Adobe Photoshop 7. These pixel counts were converted to cell counts per mm(2) using a calibration factor derived from one, two, three or all 10 images of each case/antibody combination. RESULTS: Variations in the number of labelled pixels per immunostained cell made individual calibration for each case/antibody combination necessary. Calibration based on two fields containing the most labelled pixels gave a cell count minimally higher (+5.3%) than the count based on 10-field calibration, with 95% confidence limits -14.7 to +25.3%. As TIL density could vary up to 100-fold between cases, this accuracy and precision are acceptable. CONCLUSION: The methodology described offers sufficient accuracy, precision and efficiency to quantify the density of TIL sub-populations in breast cancer using commonly available software, and could be adapted to batch processing of image files.