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Loads of suspended sediment, nutrients and pesticides discharged to the Great Barrier Reef (GBR) have increased greatly due to agricultural and urban development of the GBR catchment. As a result, in association with climate change impacts, the ecosystems of the GBR have degraded greatly in recent decades, and the decline in ecosystem health continues. Improved agricultural management practices are now being funded to reverse the decline. However the quantum of reduction in contaminant loads from individual rivers needed to reverse the decline is not known. To estimate load reductions that would reverse the decline we developed what we term Ecologically Relevant Targets (ERTs). These targets are the load reductions of sediment, nutrients and pesticides needed such that GBR lagoonal waters meet the relevant water quality guidelines for the region. The methodology proposed for setting the ERTs is described and examples of ERTs set for the Burdekin Region of the GBR are given.

The objective of this study was to evaluate the effect of pregnancy, breed, and hair coat on body temperature and sweating rate (SR) of hair sheep. St. Croix White (STX; = 9) and Dorper × STX (DRPX; = 9) ewes (3.6 yr of age) were evaluated over 4 d at 126 d of gestation (PREG) and over 4 d at 46 d postpartum (OPEN) in the shade and sun and in the morning (AM; 0900 to 1200 h) and afternoon (PM; 1300 to 1600 h) after a 20 min acclimation to each condition on each day. Data loggers recorded vaginal temperature (VT) at 10-min intervals for 96 h. Rectal temperature (RT) was measured using a digital veterinary thermometer, and respiration rate (RR) was measured as breaths per minute (bpm). Sweating rate was calculated from measured air properties passing over a shaved (300 cm) and unshaved area of the ewes' body using a portable calorimeter. Data were analyzed using GLM procedures of SAS (SAS Inst. Inc., Cary, NC) with breed, pregnancy status, sun exposure, and time of day as main effects. Mean temperature, relative humidity, temperature-humidity index, wind speed, and solar radiation on the days of data collection were 28.2°C, 82.8%, 80.3, 4.2 km/h, and 237.5 W/m, respectively. There was no difference ( > 0.10) in RT, RR, and SR between DRPX and STX ewes. The PREG ewes had lower RT ( 0.10) in RT. There was no difference in SR ( > 0.10) between the shaved and unshaved area of the ewe. The DRPX ewes had higher ( 0.10). Hair coat did not have an influence on the SR of the ewes, and PREG ewes appeared to use increased respiration as opposed to sweating to help control RT. The narrower range of body temperature, measured as VT, of PREG compared to OPEN ewes may be a protective mechanism for the developing fetus.

Despite abundant epidemiological evidence in support of aging as the primary risk factor for PD, biological correlates of a connection have been elusive. In this article, we address the following question: does aging represent biology accurately characterized as pre-PD? We present evidence from our work on midbrain dopamine neurons of aging nonhuman primates that demonstrates that markers of known correlates of dopamine neuron degeneration in PD, including impaired proteasome/lysosome function, oxidative/nitrative damage, and inflammation, all increase with advancing age and are exaggerated in the ventral tier substantia nigra dopamine neurons most vulnerable to degeneration in PD. Our findings support the view that aging-related changes in the dopamine system approach the biological threshold for parkinsonism, actively producing a vulnerable pre-parkinsonian state. © 2017 The Authors. Movement Disorders published by Wiley Periodicals, Inc. on behalf of International Parkinson and Movement Disorder Society.

The objective of this study was to investigate the direct effects of feed supplements niacin and betaine on the heat shock responses of in vitro cultured cells derived from bovine mammary and uterine tissues. First, we determined the mRNA expression profiles of the niacin receptor (GPR109A) in bovine tissues (liver, skin, uterus, udder, and ovary) and in cells derived from bovine mammary epithelium (mammary alveolar cells, MAC-T; bovine mammary epithelial cells, BMEC) and endometrium (bovine endometrial cells, BEND). We found that GPR109A was distributed in all examined tissues and cells, and the highest expression was in cells from skin and udder. Second, we evaluated the effects of niacin treatment on the mRNA abundance of heat shock proteins 70 and 27 (HSP70 and HSP27) in MAC-T, BMEC, and BEND under thermoneutral conditions and heat stress, and whether these effects were associated with alterations in the mRNA expression of prostaglandin E2 synthesis-related genes, including cyclooxygenase 1 and 2 (COX-1 and COX-2) and microsomal prostaglandin E synthase 1 and 2 (mPGES-1 and mPGES-2). Quantitative PCR data indicated that niacin suppressed HSP70 mRNA expression in BMEC and both HSP70 and HSP27 in BEND under thermoneutral conditions. Only COX-2 expression was downregulated by niacin in BMEC; other prostaglandin E2 synthesis-related genes stayed unaltered in BMEC and BEND. The mRNA abundance of HSP70, COX-1, COX-2, and mPGES-1 were elevated in niacin-treated MAC-T. During heat stress, niacin increased mRNA levels of HSP70 and HSP27 in MAC-T and HSP27 in BEND, but decreased HSP70 in BMEC. Although mPGES-2 was stimulated by niacin in BEND, the mRNA expression of prostaglandin E2 synthesis-related genes were consistent with neither HSP70 nor HSP27 expression patterns in niacin-treated BMEC and MAC-T. These data suggest that the effects of niacin on heat shock protein expression and prostaglandin E2 synthesis were not well coupled in these cells. Finally, we tested the effects of betaine treatment on viability and apoptosis in BMEC. Compared with control cultures, viability was higher in betaine-treated cells at 8 h under thermoneutral conditions and at 16 h in heat stress, and apoptotic rates were lower at 8 h. Our data support a dual role for niacin in regulating heat shock protein expression in normal and heat-shocked cells derived from mammary and uterine tissues, and positive effects of betaine in regulating mammary cell viability during heat stress.

Background: One of the most significant contributors to morbidity and death in patients with burns is infection, which accounts for 30%–75% of post-burn fatalities. Because of concerns for the development of antibiotic resistance in burn-related pathogens, the aims of this study were to identify antibiotic resistance trends for the four most common burn-related pathogens over a six-year period. Patients and Methods: The study used prospectively collected and de-identified aggregate data for all burn patients admitted to the burn unit between 2009 and 2014. The database query included patient demographics along with all data regarding cultures from any source (wound, sputum, urine, stool, blood), which included isolated pathogens and antibiotic resistance profiles. Profiles were then constructed using mean inhibitory concentration standards to determine whether each pathogen was susceptible (S), intermediate (I), or resistant (R) to each antibiotic. The aggregated data were then used to construct a heat map for the four most common pathogens and their resistance profiles over the six-year study period. Results: There were 368 patients with burns (248 with positive cultures, 120 with negative cultures) in this study. The positive culture group was significantly older (38 vs. 25 y, p

Laboratory experiments on the New Zealand freshwater mussel Echyridella menziesii were used to investigate the short-term effects (7–8 days) of food type on rates of biodeposition and benthic subst...

Inflicted burns are one of the leading causes of abuse-related fatalities in children. Between 30 and 60% of children accidentally returned to abusive homes suffer reabuse. Given the high chance for abuse recurrence and the associated morbidity/mortality, it is critical that inflicted burns are promptly identified to guide appropriate medical and child welfare management. Although previous studies proposed historical and mechanistic features using noncomparative or poorly powered data, this study utilized comparative data from a 6-year period (2009-2014) at a certified burn center along with expert analysis from Child Advocacy and Protective Services (CAPS) to provide higher level evidence supporting classical findings while elucidating new features with respect to burn severity and required interventions. A retrospective chart review of 408 pediatric burns was cross-referenced with the respective CAPS consultations to construct a multidisciplinary, deidentified database. The average age was 2.9 years (0.04-17 years) with 232 (57%) males and 330 (81%) African-Americans. CAPS investigations confirmed burn etiologies: noninflicted (346 [85%]), negligent (30 [7%]), and inflicted (32 [8%]). In comparing the three etiologies, statistical significance (P < .05) was observed for numerous variables including historical inconsistency, burn age, child welfare history, burn size and depth, distribution, concomitant injury rates, number of surgical interventions, infectious complications, and hospital length of stay. In addition to reaffirming classical features of abusive burns to fortify etiologic diagnoses, this study elucidated appreciable differences in burn severity, interventional sequelae, and burn-related complications, which will help guide medical and surgical interventions for future pediatric burn patients.

Introduction The four main prognostic models used to determine risk of burn mortality are the revised Baux (rBaux), Belgian Outcome in Burn Injury (BOBI), Abbreviated Burn Severity Index (ABSI), and quick Sequential Organ Failure Assessment (qSOFA). These models fail to factor in medical comorbidities. The ASA PS (American Society of Anesthesiologists Physical Status) is a simple scale incorporating severity of traumatic injury with comorbidities, which strongly predicts mortality in surgical patients. The purpose of this study is to determine whether the ASA PS is an adequate adjunct to measure burn severity. Methods All adults admitted to an ABA verified burn center from January 2016 to April 2019 with TBSA ≥10% who underwent surgery were reviewed. Demographics (age, gender, TBSA, race, ASA PS), vital signs (GCS, blood pressure, respiratory rate), and outcome variables (length of stay [LOS], mechanical ventilation [MV] days, and complications) were evaluated. rBaux, BOBI, ABSI, and qSOFA scores were calculated. The primary outcome was in-hospital mortality. After descriptive statistical analysis, mortality associations of the models were assessed by determining odds ratios. Firth’s logistic regression and area under the receiver operator curves determined the predictive utility of the prognostic scores. Results Of the 183 patients who fit inclusion criteria, median age was 44 years (30–57), and the majority (70%) were male. Median TBSA was 20%, 65% (n=118) had full thickness burns, 14% (n=25) had inhalation injury, and mortality was 9% (n=17). rBaux score was the best predictor of mortality (AUC=.84), ICU LOS (R2=.04), and MV days (R2=.06). For every 10-point increase in rBaux score, there was a 1.7 times increase in mortality (OR=1.7, CI 1.4–2.3, p< .00). The predicted ICU LOS increases from 2.8 to 31.4 days for the lowest and highest rBaux score quartiles. Compared to rBaux scores of 30–53, patients with scores of 54–70 had 4 times more MV days (CI 1.5–11, p< .00). The ASA PS was slightly inferior to rBaux in predicting mortality (AUC=.72), although not statistically significant (p=0.1). As ASA PS score went from I/II to III, III to IV, and IV to V/VI; mortality increased by 2.8 (OR=2.8, CI 1.5–5.5, p< .00). Conclusions rBaux is the best predictor of mortality, ICU LOS, and MV days although ASA PS also predicts mortality. Future studies should determine the combined predictive ability of ASA PS and rBaux. Applicability of Research to Practice rBaux and ASA PS scores can be used to determine risk of mortality in burn patients.

Introduction Hypoglycemic episodes are associated with worse hospital outcomes, and their incidence varies by institution. We sought to define the prevalence of hypoglycemic episodes at our burn center and determine their association with hospital outcomes. Methods We retrospectively reviewed all consecutive adult and pediatric patients admitted to our burn center from 2015 to 2019. Patient demographics and burn characteristics were recorded. The primary outcome was mortality, and secondary outcomes were total length of stay (LOS) and intensive care unit LOS (ICU LOS). All patients experiencing hypoglycemic episodes were compared to patients who did not experience hypoglycemic episodes (controls) using two-tailed t, chi-squared, and multivariate logistic and multiple linear regression analyses. Results A total of 914 patients with acute burns were admitted during the study period. Thirty-three patients (4%) experienced hypoglycemic episodes (< 60 mg/dL). Of these, 17 patients (52%) experienced a single hypoglycemic episode, while the remainder (N=16) experienced multiple hypoglycemic episodes. There were no significant differences in age, sex, or presence of inhalation injury between hypoglycemic patients and controls. Patients that experienced a hypoglycemic episode had significantly greater TBSA involvement (23% vs. 11%, p< 0.00), higher prevalence of diabetes (48% vs. 20%, p< 0.00), higher mortality (18% vs. 7%, p=0.01), longer total LOS (39 vs. 13 days, p< 0.00), and longer ICU LOS (28 vs. 4 days, p< 0.00). A single hypoglycemic episode was associated with prolonging total LOS by 19 days (p< 0.00) and ICU LOS by 18 days (p< 0.00). Hypoglycemic episodes were not associated with higher odds of mortality (OR=0.9, 95% CI 0.3–3.0, p >0.05). There were no differences in outcomes between patients with single or multiple hypoglycemic episodes. Patients with multiple hypoglycemic episodes more frequently had a history of diabetes (81% vs. 18%, p< 0.00), and worse glucose control (HbA1c, 9% vs. 7%, p=0.04) compared to patients with single episodes. Conclusions Hypoglycemic episodes were associated with longer total and ICU LOS in our study but did not portend higher mortality. Applicability of Research to Practice Quality improvement efforts to prevent hypoglycemic episodes should focus efforts on known diabetics with high HbA1c levels.

Introduction Road rash is a unique burn injury due to imbedded foreign debris and deeply seeded bacteria. Literature on this injury fails to address its unique mechanism and ways to reduce scarring and infection. Given the paucity of data, a retrospective review was performed at a single level 1 trauma center to study friction burn characteristics and outcomes from these injuries Methods We performed a systematic literature search with PubMed, Scopus, and OvidSP Medline databases using the following keywords: friction burn, traumatic tattoo, and road rash. Subsequently, we analyzed all patients treated by our ABA verified burn center for friction related injuries from January 1, 2015 to September 15, 2019 and evaluated demographics, interventions, and outcomes. Results Twenty-four pertinent articles were identified. Seven articles were included after full-text analysis. Analysis of 225 patients from these studies identified an average age of 23±7 years with 66% male. Most occurred from motor accidents (74%) with industrial (11%) and blast (10%) mechanisms accounting for the majority of the remaining cases. None of the studies recorded TBSA, but 49% were full thickness and 36% had associated injuries. Most patients (58%) required surgery. At our institution, 44 patients met inclusion criteria and the majority were young (25±18 years), men (75%), involved in motor accidents (75%), and low TBSA burns (6.3±5.6%). Over half (57%) had full thickness burns and 72% required surgery. Length of stay was 12±11 days. Of the 27% of patients with traumatic tattooing on admission, 92% successfully had all foreign debris removed by discharge. Eight (18%) patients had wound infections. Importantly, 75% of patients with wound infections had delayed evaluation by a burn practitioner; elucidating a strong and significant correlation (p< 10–5, Cramer’s V 0.84). Conclusions While road rash injuries are often small TBSA, the majority tend to be full thickness, often involve underlying critical structures and frequently require operative intervention. Significantly, patients who developed wound infections were often not evaluated by a burn provider. Applicability of Research to Practice The unique trauma of road rash causes a high degree of superficial and deep tissue injury with high risk for contamination, infection, and scarring such that rapid debridement is critical for optimal care. Burn surgeons should be consulted early as these patients have a high risk of wound infection when not consulted by wound experts.

SUMMARYA complex and poorly understood interplay between 3D genome organisation, transcription factors and chromatin state underpins cell identity. To gain a systems-level understanding of this interplay, we generated a high-resolution atlas of annotated chromatin interactions in naïve and primed human pluripotent stem cells and developed a network-graph approach to examine the atlas at multiple spatial scales. Investigating chromatin interactions as a network uncovered highly connected hubs that changed substantially in interaction frequency and in transcriptional co-regulation between pluripotent states. Small hubs frequently merged to form larger networks in primed cells, often linked by newly-formed Polycomb-associated interactions. Importantly, we identified state-specific differences in enhancer activity and interactivity that corresponded with widespread reconfiguration of transcription factor binding and target gene expression. These findings provide multilayered insights into the gene regulatory control of human pluripotency and our systems-based network approach could be applied broadly to uncover new principles of 3D genome organisation.

SUMMARYTelencephalic organoids generated from human pluripotent stem cells (hPSCs) are emerging as an effective system to study the distinct features of the developing human brain and the underlying causes of many neurological disorders. While progress in organoid technology has been steadily advancing, many challenges remain including rampant batch-to-batch and cell line-to-cell line variability and irreproducibility. Here, we demonstrate that a major contributor to successful cortical organoid production is the manner in which hPSCs are maintained prior to differentiation. Optimal results were achieved using fibroblast-feeder-supported hPSCs compared to feeder-independent cells, related to differences in their transcriptomic states. Feeder-supported hPSCs display elevated activation of diverse TGFβ superfamily signaling pathways and increased expression of genes associated with naïve pluripotency. We further identify combinations of TGFβ-related growth factors that are necessary and together sufficient to impart broad telencephalic organoid competency to feeder-free hPSCs and enable reproducible formation of brain structures suitable for disease modeling.HIGHLIGHTShPSC maintenance conditions influence outcomes in cortical organoid formationIdentification of an intermediate pluripotency state optimal for cortical organoidsFeeder support involves activation of diverse TGFβ signaling pathwaysThe organoid-promoting effects of feeders can be mimicked by a TGFβ factor mixture

Catchment degradation causing increased sediment flow is one of the key stressors facing Great Barrier Reef (GBR) habitats. Ecologically relevant targets (ERTs) for sediment and nutrient loads have been previously proposed based on seagrass light requirements, the next step is to connect these to ecological response. The overarching goal of the present work is to recommend preliminary thresholds that can be used in the development of more refined ERTs. To achieve this, we perform statistical analysis on datasets for catchment flows and sediment loads and condition of the adjacent seagrass habitat, to identify what might be the direct impacts of catchment discharge on seagrass and the associated timescales of ecological response.Our case study focuses on Cleveland Bay, which is located in the central GBR, and has important seagrass habitat that is affected by discharge from the Burdekin River. Annual monitoring of seagrass biomass and area has been undertaken since 2007. We compare these ecological time-series with data for Burdekin River annual flow and total sediment load from 2005 onwards. Annual Burdekin River flow varied by nearly 40-fold within the 2005-2018 study period, and declines in biomass and area of both subtidal and intertidal seagrasses were associated with high flows and loads from the Burdekin. Subtidal seagrasses appeared more sensitive to changes in catchment discharges than intertidal seagrasses, exhibiting a 3 year timeframe for recovery, following high annual flows and loads. Based on our results, a linear model relating change in seagrass biomass to Burdekin River metrics was used to calculate predicted thresholds below which seagrass biomass was likely to increase, and above which biomass was likely to decline. For seagrass area, a growth threshold, below which seagrass area expanded; and a decline threshold, above which seagrass area fell, were defined for annual Burdekin River flow, and sediment load. Overall these thresholds provide the first steps towards refining ERTs based on ecological condition, which can directly inform the management of the GBR to protect its iconic seagrass habitats and associated communities. The next step is to examine whether the relationship between river discharge and sediment load was the primary cause of seagrass decline.

Parkinson’s disease (PD) is a prevalent neurodegenerative disease with no approved disease-modifying therapies. Multiplications, mutations, and single nucleotide polymorphisms in the SNCA gene, encoding alpha-synuclein protein (aSyn), either cause or increase risk for PD. Intracellular accumulations of aSyn are pathological hallmarks of PD. Taken together, reduction of aSyn production may provide a disease-modifying therapy for PD. We show that antisense oligonucleotides (ASOs) reduce production of aSyn in rodent pre-formed fibril (PFF) models of PD. Reduced aSyn production leads to prevention and removal of established aSyn pathology and prevents dopaminergic cell dysfunction. In addition, we address the translational potential of the approach through characterization of human SNCA targeting ASOs that efficiently suppress the human SNCA transcript in vivo. We demonstrate broad activity and distribution of the human SNCA ASOs throughout the non-human primate brain and a corresponding decrease in aSyn cerebral spinal fluid (CSF) levels. Taken together, these data suggest that by inhibiting production of aSyn it may be possible to reverse established pathology and thus supports the development of SNCA ASOs as a potentially disease modifying therapy for PD and related synucleinopathies.SummaryAntisense oligonucleotides designed against SNCA, which are progressing to the clinic, have the potential to be a disease modifying therapeutic for Parkinson’s disease patients.

Animal models that accurately recapitulate the accumulation of alpha-synuclein (α-syn) inclusions, progressive neurodegeneration of the nigrostriatal system and motor deficits can be useful tools for Parkinson's disease (PD) research. The preformed fibril (PFF) synucleinopathy model in rodents generally displays these PD-relevant features, however, the magnitude and predictability of these events is far from established. We therefore sought to optimize the magnitude of α-syn accumulation and nigrostriatal degeneration, and to understand the time course of both. Rats were injected unilaterally with different quantities of α-syn PFFs (8 or 16 μg of total protein) into striatal sites selected to concentrate α-syn inclusion formation in the substantia nigra pars compacta (SNpc). Rats displayed an α-syn PFF quantity-dependent increase in the magnitude of ipsilateral SNpc inclusion formation at 2 months and bilateral loss of nigral dopamine neurons at 6 months. Unilateral 16 μg PFF injection also resulted in modest sensorimotor deficits in forelimb adjusting steps associated with degeneration at 6 months. Bilateral injection of 16 μg α-syn PFFs resulted in symmetric bilateral degeneration equivalent to the ipsilateral nigral degeneration observed following unilateral 16 μg PFF injection (~50% loss). Bilateral PFF injections additionally resulted in alterations in several gait analysis parameters. These α-syn PFF parameters can be applied to generate a reproducible synucleinopathy model in rats with which to study pathogenic mechanisms and vet potential disease-modifying therapies.

The transcription factor Nurr1 has been identified to be ectopically induced in the striatum of rodents expressingl-DOPA-induced dyskinesia (LID). In the present study, we sought to characterize Nurr1 as a causative factor in LID expression. We used rAAV2/5 to overexpress Nurr1 or GFP in the parkinsonian striatum of LID-resistant Lewis or LID-prone Fischer-344 (F344) male rats. In a second cohort, rats received the Nurr1 agonist amodiaquine (AQ) together withl-DOPA or ropinirole. All rats received a chronic DA agonist and were evaluated for LID severity. Finally, we performed single-unit recordings and dendritic spine analyses on striatal medium spiny neurons (MSNs) in drug-naïve rAAV-injected male parkinsonian rats. rAAV-GFP injected LID-resistant hemi-parkinsonian Lewis rats displayed mild LID and no induction of striatal Nurr1 despite receiving a high dose ofl-DOPA. However, Lewis rats overexpressing Nurr1 developed severe LID. Nurr1 agonism with AQ exacerbated LID in F344 rats. We additionally determined that inl-DOPA-naïve rats striatal rAAV-Nurr1 overexpression (1) increased cortically-evoked firing in a subpopulation of identified striatonigral MSNs, and (2) altered spine density and thin-spine morphology on striatal MSNs; both phenomena mimicking changes seen in dyskinetic rats. Finally, we provide postmortem evidence of Nurr1 expression in striatal neurons ofl-DOPA-treated PD patients. Our data demonstrate that ectopic induction of striatal Nurr1 is capable of inducing LID behavior and associated neuropathology, even in resistant subjects. These data support a direct role of Nurr1 in aberrant neuronal plasticity and LID induction, providing a potential novel target for therapeutic development.SIGNIFICANCE STATEMENTThe transcription factor Nurr1 is ectopically induced in striatal neurons of rats exhibiting levodopa-induced dyskinesia [LID; a side-effect to dopamine replacement strategies in Parkinson's disease (PD)]. Here we asked whether Nurr1 is causing LID. Indeed, rAAV-mediated expression of Nurr1 in striatal neurons was sufficient to overcome LID-resistance, and Nurr1 agonism exacerbated LID severity in dyskinetic rats. Moreover, we found that expression of Nurr1 inl-DOPA naïve hemi-parkinsonian rats resulted in the formation of morphologic and electrophysiological signatures of maladaptive neuronal plasticity; a phenomenon associated with LID. Finally, we determined that ectopic Nurr1 expression can be found in the putamen ofl-DOPA-treated PD patients. These data suggest that striatal Nurr1 is an important mediator of the formation of LID.

BCS1L encodes a homolog of the Saccharomyces cerevisiae bcs1 protein, which has a known role in the assembly of Complex III of the mitochondrial respiratory chain. Phenotypes reported in association with pathogenic BCS1L variants include growth retardation, aminoaciduria, cholestasis, iron overload, lactic acidosis and early death (GRACILE syndrome), and Björnstad syndrome, characterized by abnormal flattening and twisting of hair shafts (pili torti) and hearing problems. Here we describe two patients harbouring biallelic variants in BCS1L; the first with a heterozygous variant c.166C>T, p.(Arg56*) together with a novel heterozygous variant c.205C>T, p.(Arg69Cys) and a second patient with a novel homozygous c.325C>T, p.(Arg109Trp) variant. The two patients presented with different phenotypes; the first patient presented as an adult with aminoaciduria, seizures, bilateral sensorineural deafness and learning difficulties. The second patient was an infant who presented with a classical GRACILE syndrome leading to death at 4 months of age. A decrease in BCS1L protein levels was seen in both patients, and biochemical analysis of Complex III revealed normal respiratory chain enzyme activities in the muscle of both patients. A decrease in Complex III assembly was detected in the adult patient’s muscle, whilst the paediatric patient displayed a combined mitochondrial respiratory chain defect in cultured fibroblasts. Yeast complementation studies indicate that the two missense variants, c.205C>T, p.(Arg69Cys) and c.325C>T, p.(Arg109Trp), impair the respiratory capacity of the cell. Together, these data support the pathogenicity of the novel BCS1L variants identified in our patients.

Noradrenergic locus coeruleus (LC) neuron loss is a significant feature of mild cognitive impairment and Alzheimer's disease (AD). The LC is the primary source of norepinephrine in the forebrain, where it modulates attention and memory in vulnerable cognitive regions such as prefrontal cortex (PFC) and hippocampus. Furthermore, LC-mediated norepinephrine signaling is thought to play a role in blood-brain barrier (BBB) maintenance and neurovascular coupling, suggesting that LC degeneration may impact the high comorbidity of cerebrovascular disease and AD. However, the extent to which LC projection system degeneration influences vascular pathology is not fully understood. To address this question in vivo, we stereotactically lesioned LC projection neurons innervating the PFC of six-month-old Tg344-19 AD rats using the noradrenergic immunotoxin, dopamine-β-hydroxylase IgG-saporin (DBH-sap), or an untargeted control IgG-saporin (IgG-sap). DBH-sap-lesioned animals performed significantly worse than IgG-sap animals on the Barnes maze task in measures of both spatial and working memory. DBH-sap-lesioned rats also displayed increased amyloid and inflammation pathology compared to IgG-sap controls. However, we also discovered prominent parenchymal albumin extravasation with DBH-sap lesions indicative of BBB breakdown. Moreover, microvessel wall-to-lumen ratios were increased in the PFC of DBH-sap compared to IgG-sap rats, suggesting that LC deafferentation results in vascular remodeling. Finally, we noted an early emergence of amyloid angiopathy in the DBH-sap-lesioned Tg344-19 AD rats. Taken together, these data indicate that LC projection system degeneration is a nexus lesion that compromises both vascular and neuronal function in cognitive brain areas during the prodromal stages of AD.

Background MRI with inhaled hyperpolarized helium 3 (

In the past 12 months, many important new clinical trials in cardiology have had their first conference presentation and publication. This paper presents a constructive critical appraisal of 6 key studies. In time order of first presentation, they are CABANA, ATTR-ACT, COAPT, DECLARE, REDUCE-IT, and AUGUSTUS. For each study, the aim herein is to document and interpret the main findings, paying attention to new findings, their research context, and study limitations. These topical examples also provide methodological insights pertinent to future clinical trials research.

Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) system has emerged as an extremely useful tool for biological research and as a potential technology for gene therapy approaches. CRISPR/Cas mediated genome editing can be used to easily and efficiently modify endogenous genes in a large variety of cells and organisms. Furthermore, a modified version of the Cas9 nuclease has been developed that can be used for regulation of endogenous gene expression and labeling of genomic loci, among other applications. This chapter provides an introduction to the basis of the technology and a detail protocol for the most classic application: gene inactivation by CRISPR/Cas9 nuclease system from Streptococcus pyogenes. This workflow can be easily adapted for other CRISPR systems and applications.

Objective: A systematic review and meta-analysis was performed to summarise the state of the literature in regard to the efficacy and uses of clostridial collagenase ointment (CCO) in the burn patient. Method: A systematic review of articles available on PubMed, Scopus and OvidSP Medline was performed. Keywords used in the search process included burns, thermal injury, collagenase, enzymatic debridement, wound care. Reviews, case reports, independent abstracts, consensus and opinion papers were excluded. A meta-analysis was performed for articles fitting inclusion criteria. Results: Following screening, six relevant articles were identified for systematic review. Few studies, with limited sample sizes, argue that CCO may be an effective debriding agent. It may also accelerate wound healing and avoid the pain associated with mechanical debridement. CCO lacks antimicrobial activity but the risk of burn wound infection does not appear to be significantly different than when using silver-impregnated products. CCO is more expensive than traditional wound care products but may help halt burn depth conversion and prevent the need for surgery. Conclusion: CCO may be a safe and effective debridement agent for burn wounds with respect to decreasing wound healing time and minimising pain without increasing the risk of infection. It should be used on a case-by-case basis due to its financial cost, which may be offset by its ability to manage burns non-operatively.

Background In 1949, a French vascular surgeon, Jean Kunlin, first described a venous end-to-side anastomotic technique to address large vessel size mismatches. It was later modified by Dr. Robert Linton for end-to-end (ETE) anastomoses on the “macrovascular” scale. While multiple procedures exist for addressing moderate caliber discrepancies, few safely and reliably compensate for marked diameter differences at the microvascular level. In this study, we present the first large series adapting a modified geometric Kunlin's technique for arterial and venous ETE anastomoses in reconstructive microsurgery. Methods We reviewed 100 consecutive cases of free flap reconstructions from 2006 to 2015 utilizing a modified geometric Kunlin's technique for arterial and venous ETE anastomoses. Patient demographics and flap outcomes were analyzed. Overall flap survival rates and postoperative complications were analyzed and compared with standard values in the microsurgical literature. Results A total of 146 ETE anastomoses from 100 free flaps were analyzed. The average patient age was 51 ± 3.1 years (range: 10–84 years). Free flaps in this study were used for gender confirmation phalloplasties (20%) and reconstructing defects of the head and neck (63%), extremities (16%), and trunk (1%). There was a total of 17 takebacks including 5 for hematoma, 8 for anastomotic thromboses (6 venous), and 4 without any identifiable complication. Of these takebacks, two partial and two complete flap losses occurred. There was an 88% salvage rate for flaps requiring takebacks during the postoperative period. Overall, flap survival rate was 98% including those with complications or takebacks. Conclusion The modified Kunlin's technique is a reliable, facile, and versatile method of performing ETE anastomoses for arterial and venous vessels, especially with vessel diameter mismatches of 3:1 or greater, as well as for challenging orientations. Notably, it provides equivalent flap survival and complication rates as compared with other techniques in the microsurgical literature.

AIMS In this study, the association between multidrug resistance (MDR) and the expression of some virulence factors were evaluated in Escherichia coli strains isolated from infant faeces and fresh green vegetables. The effect of isolate origin on associated virulence factors was evaluated. In addition, genetic fingerprinting of a sample of these isolates (10 isolates from each group) was studied in order to detect any genetic relatedness among these isolates. METHODS AND RESULTS Escherichia coli isolates were divided into four groups based on their origin (human faeces or plant) and their antibiotic resistance (multiresistance or susceptible). PCR was used to investigate heat-labile and heat-stable enterotoxin genes, and four siderophore genes (aerobactin, enterobactin, salmochelin and yersiniabactin). Genetic fingerprinting of the isolates was performed using enterobacterial repetitive intergenic consensus PCR. Siderophore production was measured by a colorimetric method. Biofilm formation was evaluated by a crystal violet assay. The results of the study showed that the expression of MDR is not significantly associated with an increase in these virulence factors or with biofilm formation. However, the origin of isolates had a significant association with siderophore gene availability and consequently on the concentrations of siderophores released. Genetic fingerprinting indicated that human and plant isolates have the same clonal origin, suggesting their circulation among humans and plants. CONCLUSION Antibiotic-susceptible strains of E. coli may be as virulent as MDR strains. Results also suggest that the environment can play a potential role in selection of strains with specific virulence factors. SIGNIFICANCE AND IMPACT OF THE STUDY Antibiotic-susceptible isolates of Escherichia coli from plant or human origin can be as virulent as the multidrug resistance (MDR) ones. Genetic relatedness was detected among the isolates of plant and human origin, indicating the circulation of these bacteria among human and plants. This could imply a potential role for environmental antimicrobial resistant bacteria in human infection.

Seagrasses, flowering marine plants that form underwater meadows, play a significant global role in supporting food security, mitigating climate change and supporting biodiversity. Although progress is being made to conserve seagrass meadows in select areas, most meadows remain under significant pressure resulting in a decline in meadow condition and loss of function. Effective management strategies need to be implemented to reverse seagrass loss and enhance their fundamental role in coastal ocean habitats. Here we propose that seagrass meadows globally face a series of significant common challenges that must be addressed from a multifaceted and interdisciplinary perspective in order to achieve global conservation of seagrass meadows. The six main global challenges to seagrass conservation are (1) a lack of awareness of what seagrasses are and a limited societal recognition of the importance of seagrasses in coastal systems; (2) the status of many seagrass meadows are unknown, and up-to-date information on status and condition is essential; (3) understanding threatening activities at local scales is required to target management actions accordingly; (4) expanding our understanding of interactions between the socio-economic and ecological elements of seagrass systems is essential to balance the needs of people and the planet; (5) seagrass research should be expanded to generate scientific inquiries that support conservation actions; (6) increased understanding of the linkages between seagrass and climate change is required to adapt conservation accordingly. We also explicitly outline a series of proposed policy actions that will enable the scientific and conservation community to rise to these challenges. We urge the seagrass conservation community to engage stakeholders from local resource users to international policy-makers to address the challenges outlined here, in order to secure the future of the world’s seagrass ecosystems and maintain the vital services which they supply. Electronic supplementary material The online version of this article (10.1007/s13280-018-1115-y) contains supplementary material, which is available to authorised users.

Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) system has emerged as an extremely useful tool for biological research and as a potential technology for gene therapy approaches. CRISPR/Cas mediated genome editing can be used to easily and efficiently modify endogenous genes in a large variety of cells and organisms. Furthermore, a modified version of the Cas9 nuclease has been developed that can be used for regulation of endogenous gene expression and labeling of genomic loci, among other applications. This chapter provides an introduction to the basis of the technology and a detail protocol for the most classic application: gene inactivation by CRISPR/Cas9 nuclease system from Streptococcus pyogenes. This workflow can be easily adapted for other CRISPR systems and applications.

Introduction: In desert climates, high temperatures can cause heat stress on dairy farms. Aviable alternative to this problem is to develop a network of water pipes connected to heat exchangers (HEs) that operate as a conductive cooling system. Objective: To propose a water supply system connected to a series of HEs, installed under bedding in a dairy barn freestall system, and analyze heat transfer along a large-scale water pipe network. Methodology: The EPANET water quality module was used to design the system. Temperature predictions were validated with experimental data from a network of four HE pipes installed under the bedding. A network was then designed to supply water to 1 000 HEs in order tocalculate the efficiency of the system for a real dairy farm. Results: Insulations with values of 0.095 and 0.0475 W∙m-1∙°C-1 increased the cooling capacity of water-carrying pipes by 7 and 12 %, respectively. Increasing the flow rates from 1 to 7 L∙min-1 increased the cooling capacity; however, the cooling efficiency decreased by at least 12 %. Study limitations: A validation was performed on a section of the pipe network and a simulation for the entire network. Originality:V EPANET was implemented for a mass and heat balance in a pipe network supplying water to HEs. Conclusion: The conductive cooling system is viable in areas with a desert climate and its efficiency depends on the level of thermal insulation and flow in the pipe network.

Introduction Bioelectric Impedance Analysis (BIA) is a rapid, non-invasive, and inexpensive technology based on electrical conductivity. BIA assesses body composition, fluid shifts, and phase angle, an electrical force vector where smaller values suggest cellular injury. Our objective was to use BIA to longitudinally track the clinical status of burn patients. We hypothesized that BIA would detect progressive decreases in muscle mass throughout the patient’s hospital course and that low phase angle values would correlate with severity of injury. Methods A cohort study of 10 patients from January 1, 2020 to March 13, 2020 was performed at an ABA-verified burn center. Patient and burn characteristics and laboratory data were collected. BIA measurements were performed daily for the first 10 days of admission and then twice weekly until discharge. The primary outcome was to detect changes in body composition. The secondary objectives were to detect differences between low and high-risk patients in terms of water composition and phase angle. Patients with APACHE II > 15, measured at burn unit admission, were considered high risk for burn injury related morbidity and mortality. Results BIA detected a statistically significant negative correlation between time spent hospitalized and leg lean mass (LM) (r2=0.56, P< 0.0001), right arm LM (r2=0.52, P< 0.0001) and left leg LM (r2=0.57, P< 0.0001), and positive correlation between body fat mass (BFM) and time spent hospitalized (r2=0.50, P=0.0004). Water composition (volume of extracellular water (ECW) per total body water (TBW)) negatively correlated with low-risk patients: right arm (r2=0.51, P< 0.0001), left arm (0.71, P< 0.001), thorax (0.66, P< 0.0001), right leg (0.74, P< 0.0001), left leg (0.35, P=0.002). Full body phase angle increased with low-risk patients over their hospital course (r2=0.62, P< 0.0001), while phase angle decreased with high-risk patients (r2=0.71, P=0.0006). Full body phase angle differentiated high risk patients (P< 0.0001), and phase angle of thorax differentiated between patients with and without inhalation injury (P=0.002). Conclusions Our study demonstrates that BIA measures changes in body composition and fluid shifts, identifies inhalation injury, and correlates with severity of injury in hospitalized burn patients. This pilot study included a limited number of participants with varying anatomic injuries presenting unique measurement challenges. Regardless, our preliminary data justifies a larger prospective study to confirm these results and correlate them with clinical outcomes.

Prevalent in approximately 20% of the worldwide human population, the rs6265 (also called 'Val66Met') single nucleotide polymorphism (SNP) in the gene for brain-derived neurotrophic factor (BDNF) is a common genetic variant that can alter therapeutic responses in individuals with Parkinson's disease (PD). Possession of the variant Met allele results in decreased activity-dependent release of BDNF. Given the resurgent worldwide interest in neural transplantation for PD and the biological relevance of BDNF, the current studies examined the effects of the rs6265 SNP on therapeutic efficacy and side-effect development following primary dopamine (DA) neuron transplantation. Considering the significant reduction in BDNF release associated with rs6265, we hypothesized that rs6265-mediated dysfunctional BDNF signaling contributes to the limited clinical benefit observed in a subpopulation of PD patients despite robust survival of grafted DA neurons, and further, that this mutation contributes to the development of aberrant graft-induced dyskinesias (GID). To this end, we generated a CRISPR knock-in rat model of the rs6265 BDNF SNP to examine for the first time the influence of a common genetic polymorphism on graft survival, functional efficacy, and side-effect liability, comparing these parameters between wild-type (Val/Val) rats and those homozygous for the variant Met allele (Met/Met). Counter to our hypothesis, the current research indicates that Met/Met rats show enhanced graft-associated therapeutic efficacy and a paradoxical enhancement of graft-derived neurite outgrowth compared to wild-type rats. However, consistent with our hypothesis, we demonstrate that the rs6265 genotype in the host rat is strongly linked to development of GID, and that this behavioral phenotype is significantly correlated with neurochemical signatures of atypical glutamatergic neurotransmission by grafted DA neurons.

Native freshwater mussels (Bivalvia: Unionoida) can represent a large portion of benthic biomass, but their functional role is still poorly understood. We sampled Echyridella menziesii (Hyriidae) in six lakes from two regions of the North Island of New Zealand to develop general allometric relationships for predicting filtration, excretion and biodeposition rates. Experimental containers were set up in the field (no-flow conditions) to measure filtration and biodeposition using natural phytoplankton communities, and excretion in filtered lake water. Filtration rates were 0.02–1.3 L mussel−1 h−1 and increased with increasing mussel size (R2 = 0.13, P = 0.023). Stable isotope analysis suggests that mussels also assimilate food from non-planktonic origins. Nitrogen excretion rates were 4–50 μg N mussel−1 h−1 and increased with mussel size (R2 = 0.70, P 0.3). In contrast, phosphorus excretion rates did not vary with mussel size, and were much lower in Rotorua than Waikato lakes (2 vs 5 μg P mussel−1 h−1). The reason for this regional difference is unclear, but suggests that mussels could contribute different N:P ratios to nearshore nutrient budgets in different types of lakes. Biodeposits represented 50–70% of the N, and 25–70% of the P recycled. Echyridella filters and excretes nutrients at rates similar to those measured in North American and European mussels (Unionidae, Margaritiferidae). Mussels could be important contributors of nutrients in areas where they are abundant, and their inclusion into nutrient budget models could improve the predictions of nearshore nutrient fluxes in lakes and reservoirs.

Betaine (BET), a natural, organic osmolyte, improves cellular efficiency by acting as a chaperone, refolding denatured proteins. To test if dietary BET reduced the effect of heat stress (HS) in lactating dairy cows, multiparous, lactating Holstein cows (n=24) were blocked by days in milk (101.4±8.6 d) and randomly assigned to 1 of 3 daily intakes of dietary BET: the control (CON) group received no BET, mid intake (MID) received 57mg of BET/kg of body weight, and high dose (HI) received 114mg of BET/kg of body weight. Cows were fed twice daily and BET was top-dressed at each feeding. Cows were milked 2 times/d and milk samples were taken daily for analysis. Milk components, yield, feed intake, and water intake records were taken daily. Rectal temperature and respiration rate were taken 3 times/d at 0600, 1400, and 1800h. Cows were housed in environmentally controlled rooms and were allowed acclimation for 7d at thermoneutral (TN) conditions with a mean temperature-humidity index of 56.6. Cows were then exposed to 7d of TN followed by 7d of HS represented by a temperature-humidity index of 71.5 for 14d. This was followed by a recovery period of 3d at TN. Dietary BET increased milk yield during the TN period. No differences were found between BET and CON in total milk production or milk composition during HS. The increase in water intake during HS was not as great for cows fed BET compared with controls. The cows on CON diets had higher p.m. respiration rate than both MID and HI BET during HS, but lower rectal temperature compared with BET. No difference was found in serum glucose during TN, but cows given HI had elevated glucose levels during HS compared with CON. No differences were found in serum insulin levels between CON and BET but an intake by environment interaction was present with insulin increasing in HI-treated lactating dairy cows during HS. The heat shock response [heat shock protein (HSP) 27 and HSP70] was upregulated in bovine mammary epithelial cells in vitro. Blood leukocyte HSP27 was downregulated at the HI dose under TN conditions and HSP70 was upregulated at the HI dose and this effect was increased by HS. No effect was seen with the MID dose with HSP27 or HSP70. The lack of effect of BET at MID may be associated with uptake across the gut. We conclude that BET increased milk production under TN conditions and was associated with reduced feed and water intake and slightly increased body temperatures during HS of cows fed BET. The effect of BET on milk production was lost during HS with HI BET, whereas serum glucose levels increased during HS.

Predictions of invasion risk for seven non-indigenous fish species, ecological impact scores for individual species, and lake conservation rankings were linked to develop Invasion Risk Impact (IRI) and Lake Vulnerability (LV) indices that help identify New Zealand lakes most at risk of loss of conservation value from potential multi-species invasions. Species-specific IRI scores (the product of predicted invasion risk and species impact) highlighted Eurasian perch (Perca fluviatilis) and the brown bullhead (Ameiurus nebulosus), as the species most likely to spread and cause ecological harm in lakes. For 3431 lakes >1 ha throughout New Zealand, total IRI tended to be highest for lowland riverine and dune lakes most of which are already colonized by multiple invasive fish species. The LV index indicated that lakes most at risk of loss of conservation value from invasive fish impacts were predominantly (i) in the northern half of the North Island where several uncommon lake types occur, and (ii) along the west coast of the South Island where conservation value is often greater, largely because of low catchment modification. The IRI and LV indices can be used to assist with setting priorities for surveillance monitoring, advocacy, and response planning targeted at preventing the establishment of invasive fish in moderate-to-high value lakes most susceptible to ecological impacts. Both indices can be adapted to accommodate alternative impact and conservation scoring systems, providing a flexible tool for local- and national-scale assessments of lake vulnerability to fish invasion impacts. Copyright © 2016 John Wiley & Sons, Ltd.

Consideration of the need and opportunities for restoration of freshwater lakes in New Zealand requires individualised approaches that account for lake type, invasive species and food web structure.

Defining goals and objectives is a critical component of adaptive management of natural resources because they provide the basis on which management strategies can be designed and evaluated. The aims of this study are (i) to apply and test a collaborative method to elicit goals and objectives for inshore fisheries and biodiversity in the coastal zone of a regional city in Australia; (ii) to understand the relative importance of management objectives for different community members and stakeholders; and (iii) to understand how diverse perceptions about the importance of management objectives can be used to support multiple-use management in Australia's iconic Great Barrier Reef. Management goals and objectives were elicited and weighted using the following steps (i) literature review of management objectives, (ii) development of a hierarchy tree of objectives, and (iii) ranking of management objectives using survey methods. The overarching goals identified by the community group were to (1) protect and restore inshore environmental assets; (2) improve governance systems; and (3) improve regional (socio-economic) well-being. Interestingly, these goals differ slightly from the usual triple-bottom line objectives (environmental, social and economic) often found in the literature. The objectives were ranked using the Analytical Hierarchical Process, where a total of 141 respondents from industry, government agencies, and community from across Queensland State undertook the survey. The environment goal received the highest scores, followed by governance and lastly well-being. The approach to elicit and rank goals and objectives developed in this study can be used to effectively support coastal resource management by providing opportunities for local communities to participate in the setting of regional objectives. (C) 2016 Elsevier Ltd. All rights reserved.

Invertebrates inhabiting marine and freshwater ecosystems make important contributions to global biodiversity and provide significant services that have cascading effects across ecosystems. However, this group is grossly under-represented in assessments of conservation status and often neglected in targeted aquatic conservation efforts. In global assessments of 7857 freshwater invertebrates and 2864 marine invertebrates, 30–34% were considered Data Deficient highlighting the paucity of information for making such assessments. Of the invertebrate groups that could be assessed, those with poor dispersal abilities and high local endemism, such as many gastropods, crayfish and mussels, are the most threatened. Springs and subterranean hydrological systems support the highest proportions of threatened freshwater species, while in marine environments coral reefs, lagoons and anchialine systems are particularly vulnerable. Key agents of biodiversity decline in aquatic ecosystems are water pollution, overexploitation and harvesting, habitat degradation and destruction, alien invasive species, and climate change. Effects of dams and water management along with pollution from urban, agricultural and forestry sources are the main threats in freshwater ecosystems, whereas a broad range of factors have impacts on marine invertebrates, including biological resource use. Significant impediments facing conservation of aquatic invertebrates are limited knowledge of their diversity, the need for broadscale actions to account for connectivity within and across ecosystems, lack of political will and investment, and the prospect that conditions may get worse before they improve, possibly not in time to save some already highly imperilled invertebrate species from extinction.

The objective of this study was to determine the pharmacokinetics of single- and multi-dose ceftiofur crystalline-free acid (CCFA) administered subcutaneously at a dose of 13.2 mg/kg to 12 neonatal foals 1-3 days of age. Six foals received a single subcutaneous dose, while 6 additional foals received 4 doses of CCFA at 48-h intervals. Blood samples were collected at pre-determined times following drug administration, and plasma concentrations of ceftiofur free acid equivalents (CFAE) were measured using high-performance liquid chromatography. Following single-dose administration of CCFA, the mean ± standard deviation maximum observed plasma concentration was 3.1 ± 0.6 μg/mL and observed time to maximal plasma concentration was 14.0 ± 4.9 h. Following multi-dose administration of CCFA, the mean ±standard deviation times above CFAE concentrations of ≥0.5 μg/mL and ≥2.0 μg/mL were 192.95 ± 15.86 h and 78.80 ± 15.31 h, respectively. The mean ± standard deviation area under the concentration vs time curve (AUC0→∝ ) was 246.2 ± 30.7 h × μg/mL and 172.7 ± 27.14 h × μg/mL following single- and multi-dose CCFA administrations, respectively. Subcutaneous administration of CCFA at 13.2 mg/kg in neonatal foals was clinically well- tolerated and resulted in plasma concentrations sufficient for the treatment of most bacterial pathogens associated with neonatal foal septicemia. Multi-dose administration of four doses at dosing interval of 48 h between treatments maintains appropriate therapeutic concentrations in neonatal foals.

Background Axon viability is critical for maintaining neural connectivity, which is central to neural functionality. Many neurodegenerative diseases (e.g., Parkinson’s disease (PD) and Alzheimer’s disease) appear to involve extensive axonal degeneration that often precedes somatic loss in affected neural populations. Axonal degeneration involves a number of intracellular pathways and characteristic changes in axon morphology (i.e., swelling, fragmentation, and loss). New method We describe a relatively simple set of methods to quantify the axonal degeneration using the 6-hydroxydopamine neurotoxin model of PD in rats and a colchicine-induced model in primary rat neurons. Specifically, approaches are described that use the spaceballs stereological probe for tissue sections and petrimetrics stereological probe for cultured neurons, and image analysis techniques in both tissue sections and cultured neurons. Results These methods provide a mechanism for obtaining quantitative and semi-quantitative data to track the extent of axonal degeneration and may prove useful as outcome measures in studies aimed at preventing or slowing axonal degeneration in disease models. Comparison with existing methods Existing methods of quantification of axonal degeneration use densitometry and manual counts of axonal projections, but they do not utilize the random, unbiased systematic sampling approaches that are characteristic of stereological methods. The ImageJ thresholding analyses described here provide a descriptive method for quantifying the state of axonal degeneration. Conclusions These methods provide an efficient and effective means to quantify the extent and state of axonal degeneration in animal tissue and cultured neurons and can be used in other models for the same purposes.

Current reconstructive approaches to large craniofacial skeletal defects are often complicated and challenging. Critical-sized defects are unable to heal via natural regenerative processes and require surgical intervention, traditionally involving autologous bone (mainly in the form of nonvascularized grafts) or alloplasts. Autologous bone grafts remain the gold standard of care in spite of the associated risk of donor site morbidity. Tissue engineering approaches represent a promising alternative that would serve to facilitate bone regeneration even in large craniofacial skeletal defects. This strategy has been tested in a myriad of iterations by utilizing a variety of osteoconductive scaffold materials, osteoblastic stem cells, as well as osteoinductive growth factors and small molecules. One of the major challenges facing tissue engineers is creating a scaffold fulfilling the properties necessary for controlled bone regeneration. These properties include osteoconduction, osteoinduction, biocompatibility, biodegradability, vascularization, and progenitor cell retention. This review will provide an overview of how optimization of the aforementioned scaffold parameters facilitates bone regenerative capabilities as well as a discussion of common osteoconductive scaffold materials.

We sampled natural and reconstructed side-arms during different stages of hydrological connectivity with a large floodplain river in northern New Zealand, to determine whether re-establishment of connectivity would be an effective strategy for restoring plankton communities in former side-arms. Connectivity between side-arms and the river was moderated by water level and influenced flow rates and closure of inlets and outlets. Physicochemical conditions were more strongly related to the connectivity phase than to habitat type (river, natural or reconstructed side-arm), except during low connectivity when natural side-arms in particular were characterised by higher ammonium (NH4-N) and total phosphorus (P) concentrations, as well as specific conductivity. Dissolved reactive phosphorus (PO4-P), water temperature, conductivity and dissolved oxygen were identified as explanatory variables of phytoplankton and zooplankton community composition, which along with total nitrogen (phytoplankton) or total suspended solids (zooplankton) explained 44–52% of variation. Phytoplankton community composition and the abundance of several dominant or discriminatory taxa were affected by connectivity but not habitat type, whereas habitat and connectivity both had significant effects on zooplankton communities and abundances of the cladoceran Bosmina meridionalis. Significant interactions between connectivity and paired habitat types occurred for abundances of the diatom Asterionella, the cryptophyte Cryptomonas, the rotifer Synchaeta oblonga and cyclopoid copepods, reflecting differential responses to connectivity among habitats by these taxa. Overall, these results underscore the importance of hydrological connectivity between side-arms and rivers in moderating plankton community composition, and highlight unpredictable trajectories of community development and alternative transient states that can occur soon after side-arm reconnection. Copyright © 2016 John Wiley & Sons, Ltd.

Summary Heterogeneity of lateral habitats along large rivers can enhance rates of ecological processes such as nutrient transformations and contaminant sequestration. Consequently, construction or reconnection of hydraulic retention zones such as side-arms is being widely promoted in river restoration projects, although limited information is available to quantify the benefits to water quality. We sampled natural side-arms and a reconstructed side-arm at different phases of hydrological connectivity on a large floodplain river in northern New Zealand to quantify changes to water quality within and between side-arms, and to determine changes associated with reconnection. Sampling of seven side-arm outlets during low flow indicated reduced NO3-N concentrations but elevated conductivity and total suspended sediment (TSS) compared with the river. Longitudinal changes within disconnected side-arms were most pronounced at outlet and/or mid-arm locations where total nitrogen and NO3-N declined relative to inlets, while TSS and conductivity increased. Concentrations of dissolved oxygen were also markedly reduced in side-arms during disconnection. The natural and reconstructed side-arms displayed different temporal responses to connectivity, with NH4-N and TSS in particular becoming elevated along the natural side-arm during low connectivity, while TSS declined and PO4-P increased along the reconstructed side-arm. These temporal differences were likely caused by (i) differences in side-arm age (time since formation) affecting substrate composition, riparian vegetation and hydrological pathways, and (ii) bioturbation by large benthic-feeding invasive fish which became trapped in natural side-arms during disconnection. These results suggest potential for side-arm reconstruction to influence water quality at the local scale if (i) large invasive fish that mobilise bottom sediments can be excluded at times of disconnection, and (ii) side-arms are maintained to ensure transformation and attenuation processes persist over time and continue to provide water quality benefits.

A trial was performed to assess the effect of evaporative pads on core body temperature (CBT) and lying behavior of lactating Holstein cows housed in cross-ventilated freestall facilities in a humid environment. This trial was undertaken in 2 barns equipped with (EP) or without (NP) evaporative pads. Each facility had 4 pens, 1 baffle/pen, and a nominal width of 122 m. Stocking density was higher (123.4 vs. 113.1%) and freestalls were slightly shorter (2.3 vs. 2.4 m) and narrower (1.16 vs. 1.21 m) in EP compared with NP barns. In each pen, lying behavior of 20 cows was monitored using electronic data loggers that recorded at 1-min intervals. A subset (n=14) of these cows within each pen were also fitted with temperature loggers attached to blank controlled intravaginal drug release devices to determine CBT every 5 min. Ambient conditions were collected every 15 min. Individual cow lying duration and lying bouts were assessed for each cow, as well as time spent standing and CBT within the following categories: CBT38.6°C, and CBT38.6,38.9,39.2,39.4, and39.7°C. These variables were analyzed using pen as the experimental unit, with cow and day as additional random effects. The average maximum ambient conditions over the 9 d were 25°C and 78.74% relative humidity. No differences were observed in lying duration and number of lying bouts over the 9-d period, with overall means of 696±31 min/d and 12.6±0.5 bouts/d. The EP cows spent 170 min/d longer with a CBT38.6°C and 107 min/d less with CBT39.2°C than did NP cows. Cooling with evaporative pads tended to increase time spent lying with a CBT8.6°C and lying bouts/d for EP cows versus NP cows. Results from this trial show that even under mild heat stress, evaporative cooling in cross-ventilated facilities can decrease CBT and tended to increase lying time.

Photodynamic therapy (PDT) is an established treatment option for low-risk basal cell carcinoma (BCC). BCC is the most common human cancer and also a convenient cancer in which to study PDT treatment. This review clarifies challenges to researchers evident from the clinical use of PDT in BCC treatment. It outlines the context of PDT and how PDT treatments for BCC have been developed hitherto. The sections examine the development of systemic and subsequently topical photosensitizers, light delivery regimens, and the use of PDT in different patient populations and subtypes of BCC. The outcomes of topical PDT are discussed in comparison with alternative treatments, and topical PDT applications in combination and adjuvant therapy are considered. The intention is to summarize the clinical relevance and expose areas of research need in the BCC context, ultimately to facilitate improvements in PDT treatment.

Implementing management actions to achieve environmental outcomes requires defining and quantifying ecological targets, but this is a complex challenge, and there are few examples of how to quantitatively set them in complex dynamic marine ecosystems. Here we develop a methodology to devise ‘desired state’ for tropical seagrasses in Cleveland Bay, northern Australia, in the Great Barrier Reef World Heritage Area. Analysis of diverse species assemblages was used to define seagrass communities as indicators of the region’s ecological value. Multivariate regression trees assigned 8000 observations of species presence/absence and habitat characteristics from 2007 to 2017 into seven community types. Generalized Linear Models were used to assess annual variation in above-ground biomass of each seagrass community. Reference subsets of the data expressing high biomass and spatial extent were identified, and desired state was defined as the mean and 95% confidence intervals. This approach rests on the assumption that seagrass resilience and its ecosystem services are met when the diverse seagrass communities reach desired state. This method required a data set that spanned a range in seagrass conditions, but which may have been compromised by a history of pressures. Our method for defining desired state provides evidence-based targets that can be used within an adaptive management framework that prioritises and implements management actions.

Seagrass ecosystems are increasingly subjected to multiple interacting stressors, making the consequent trajectories difficult to predict. Here, we present a new process-based model of seagrass decline in response to cumulative light and temperature stress. The model is calibrated to laboratory datasets for Great Barrier Reef seagrasses using Bayesian inference. Our model, which is fit to both physiological and morphological data, supports the hypothesis that physiological carbon loss rate controls the shoot density decline rate of seagrasses. The model predicts the time to complete shoot loss, and a new, generalisable, cumulative stress index that indicates the potential seagrass shoot density decline based on the time period of cumulative stress. All model predictions include uncertainty estimates based on uncertainty in the model fit to the data. The calibrated model is packaged into a computer program that can forecast the potential declines of seagrasses due to cumulative light and temperature stress.

Numerous genes, and alterations in their expression, have been identified as risk factors for developing levodopa-induced dyskinesia (LID). However, our understanding of the complexities of molecular changes remains insufficient for development of clinical treatment. In the current study we used gene array, in situ hybridization, immunohistochemistry, and microdialysis to provide a unique compare and contrast assessment of the relationship of four candidate genes to LID, employing three genetically distinct rat strains (Sprague-Dawley (SD), Fischer-344 (F344) and Lewis-RT.1) showing differences in dyskinesia susceptibility and ‘first-ever LID’ versus ‘chronic LID’ expression in subjects displaying equal dyskinesia severity. In these studies, rat strains were easily distinguishable for their LID propensity with: 1) a majority of SD rats expressing LID (LID+) and a subset being resistant (LID-); 2) all F344 rats readily developing (LID+); and 3) all Lewis rats being LID-resistant (LID-). Following chronic levodopa, LID+ SD rats showed significant increases in candidate gene expression: Nr4a2/(Nurr1) > > Trh > Inhba = Fosb. However, SD rats with long-standing striatal dopamine (DA) depletion treated with first-ever versus chronic high-dose levodopa revealed that d espite identical levels of LID severity: 1) Fosb and Nurr1 transcripts but not protein were elevated with acute LID expression; 2) FOSB/ΔFOSB and NURR1 proteins were elevated only with chronic LID; and 3) Trh transcript and protein were elevated only with chronic LID. Strikingly, despite similar levodopa-induced striatal DA release in both LID-expressing F344 and LID-resistant Lewis rats, Fosb, Trh, Inhba transcripts were significantly elevated in both strains; however, Nurr1 mRNA was significantly increased only in LID+ F344 rats. These findings suggest a need to reevaluate currently accepted genotype-to-phenotype relationships in the expression of LID, specifically that of Fosb, a transcription factor generally assumed to play a causal role, and Nurr1, a transcription factor that has received significant attention in PD research linked to its critical role in the survival and function of midbrain DA neurons but who's striatal expression, generally below levels of detection, has remained largely unexplored as a regulator of LID. Finally these studies introduce a novel ‘model’ (inbred F344 vs inbred Lewis) that may provide a powerful tool for investigating the role for ‘dyskinesia-resistance’ genes downstream of ‘dyskinesia-susceptibility’ genes in modulating LID expression, a concept that has received considerably less attention and offers a new ways of thinking about antidyskinetic therapies.

Introduction Hands are the most commonly burned body part, and are prone to developing limited range of motion and scar contractures, which benefit from aggressive occupational therapy (OT). Many burn survivors are vulnerable with limited resources and poor follow up. The purpose of this study is to determine outpatient OT referral and compliance rates in our patient population. Methods All patients with hand burns admitted to a single ABA verified burn center from January 2015 to May 2016 were reviewed. Demographics (living situation, substance use, psychiatric illness), TBSA, interventions (type/number of surgeries), outcomes (length of stay [LOS]), and OT outcomes (inpatient and outpatient treatment, reason for discontinuation) were evaluated. Results Sixty-one patients met inclusion criteria. Mean age was 33±18 years. In terms of living situation, 17% (n=10) were homeless, and 3% (n=2) were incarcerated. A documented history of alcohol abuse was present in 13% (n=8), 23% (n=14) had positive urine toxicology screens, and 15% (n=9) had diagnosed psychiatric illness. Mean TBSA was 10%±6. Half (N=30, 51%) were managed non-operatively. At discharge, 30% (n=18) had normal hand function and did not meet criteria for outpatient hand therapy. Over 70% (n=43) received outpatient referrals, but only 44% (n=19) of those referred reliably returned for outpatient OT. Despite having outpatient OT referrals, 33% (n=14) did not present to outpatient therapy after repeats attempts of contact, and 67% (n=29) of those with referrals only came to one visit. Of those that did not complete therapy, most (n=14, 58%) were lost to follow up and unable to be contacted. Other reasons for not completing therapy within our system were out-of-network insurances requiring follow-up outside our hospital (36%) and incarceration (6%). Conclusions At our center, nearly 1 in 3 patients with hand burns had excellent function upon discharge and did not require outpatient therapy. However, when patients are referred for outpatient therapy, many do not show up or maintain reliable compliance. Many patients are simply lost to follow up. Applicability of Research to Practice While occupational therapy remains an effective and viable option for hand rehabilitation, further efforts must be aimed at providing patients mechanisms and education for achieving reliable outpatient follow up.

Introduction Sparse data exists on functional outcomes of patients receiving outpatient therapy after admission with hand burns. The purpose of our study is to evaluate the effectiveness of long-term outpatient occupational therapy (OT) on hand joint range of motion (ROM), activities of daily living (ADL) status, and pain after burn injury. Methods All patients with hand burns admitted to a single ABA verified burn center from January 2015 to May 2016 with properly documented outpatient OT follow up were included. Demographics (TBSA, hand dominance, mechanism), interventions (time-to-surgery, procedures), and long-term outcomes (further procedures, pain, ROM, contractures, scars, ADL) were evaluated. The effect of patient demographics and interventions on outcomes were evaluated with descriptive statistics and multivariate logistic regression. Results Of 61 patients with hand burns, 43 were referred for outpatient therapy, but only 31% (n=19) consistently presented for follow up. Mean age was 37±14 years, 74% (n=14) were male, and mean TBSA was 12%±17. Surgical management was required in 63% (n=12), and the mean time-to-surgery was 16±37 days from injury. Contractures occurred in 16% (n=3), 11% (n=2) had hypertrophic scarring, and 21% (n=4) developed both contractures and hypertrophic scarring. Of these patients, 21% (n=4) required further surgical intervention, including 3 contracture releases and 1 triamcinolone injection. Mean OT follow up was 16±14 weeks. As of the last OT note, 84% (n=16) had independent ADL function, 16% (n=3) required assistance, and none were poorly functioning. Likewise, 47% (n=9) had normal ROM, 53% (n=10) were within functional limits, and none had poor ROM. 79% (n=15) demonstrated improvement of ADL function, and 84% (n=16) had improvement of ROM. At OT intake, 68% (n=13) reported pain as a major limitation, but by end of therapy, only 21% (n=4) were limited by pain. Those with contractures or hypertrophic scars were 9.9 times less likely to have improvement in ADL status (p=.03, RR 9.9, CI 1.3–67). Conclusions Most patients referred for hand therapy after burn injury return to functional independence and have functional ROM. Applicability of Research to Practice Referral to and compliance with dedicated long-term hand therapy leads to improvements to ADL and ROM in patients admitted with hand burn injuries.