Zonal image analysis of tumour vascular perfusion, hypoxia, and necrosis

A number of laboratories are utilising both hypoxia and perfusion markers to spatially quantify tumour oxygenation and vascular distributions, and scientists are increasingly turning to automated image analysis methods to quantify such interrelationships. In these studies, the presence of regions of necrosis in the immunohistochemical sections remains a potentially significant source of error. In the present work, frozen MCa-4 mammary tumour sections were used to obtain a series of corresponding image montages. Total vessels were identified using CD31 staining, perfused vessels by DiOC7 staining, hypoxia by EF5/Cy3 uptake, and necrosis by haematoxylin and eosin staining. Our goal was to utilise image analysis techniques to spatially quantitate hypoxic marker binding as a function of distance from the nearest blood vessel. Several refinements to previous imaging methods are described: (1) hypoxia marker images are quantified in terms of their intensity levels, thus providing an analysis of the gradients in hypoxia with increasing distances from blood vessels, (2) zonal imaging masks are derived, which permit spatial sampling of images at precisely defined distances from blood vessels, as well as the omission of necrotic artifacts, (3) thresholding techniques are applied to omit holes in the tissue sections, and (4) distance mapping is utilised to define vascular spacing

Acoustic Imaging Evaluation of Juvenile Salmonid Behavior in the Immediate Forebay of the Water Temperature Control Tower at Cougar Dam, 2010

This report presents the results of an evaluation of juvenile Chinook salmon (Oncorhynchus tshawytscha) behavior at Cougar Dam on the south fork of the McKenzie River in Oregon in 2010. The study was conducted by the Pacific Northwest National Laboratory for the U.S. Army Corps of Engineers (USACE). The overall goal of the study was to characterize juvenile salmonid behavior and movement patterns in the immediate forebay of the Water Temperature Control (WTC) tower of the dam for USACE and fisheries resource managers use in making decisions about bioengineering designs for long-term structures and/or operations to facilitate safe downstream passage for juvenile salmonids. We collected acoustic imaging (Dual-Frequency Identification Sonar; DIDSON) data from March 1, 2010, through January 31, 2011. Juvenile salmonids (hereafter, called 'fish') were present in the immediate forebay of the WTC tower throughout the study. Fish abundance index was low in early spring (<200 fish per sample-day), increased in late April, and peaked on May 19 (6,039 fish). A second peak was observed on June 6 (2904 fish). Fish abundance index decreased in early June and remained low in the summer months (<100 fish per sample-day). During the fall and winter, fish numbers varied with a peak on November 10 (1881 fish) and a minimum on December 7 (12 fish). A second, smaller, peak occurred on December 22 (607 fish). A univariate statistical analysis indicated fish abundance index (log10-transformed) was significantly (P<0.05) positively correlated with forebay elevation, velocity over the WTC tower intake gate weirs, and river flows into the reservoir. A subsequent multiple regression analysis resulted in a model (R2=0.70) predicting fish abundance (log-transformed index values) using two independent variables of mean forebay elevation and the log10 of the forebay elevation range. From the approximate fish length measurements made using the DIDSON imaging software, the average fish length during early spring 2010 was 214 {+-} 86 mm (standard deviation). From May through early November, the average fish length remained relatively consistent (132 {+-} 54 mm), after which average lengths increased to 295 {+-} 148 mm for mid-November though early December. From mid-December through January the average fish length decreased to 151 {+-} 76 mm. Milling in front of the WTC tower was the most common fish behavior observed throughout the study period. Traversing along the front of the tower, east-to-west and west-to-east, was the next common behavior. The percentage of fish events showing movement from the forebay to the tower or from the tower to the forebay was generally low throughout the spring, summer, and early fall (0 to 30% for both directions combined, March through early November). From mid-November 2010 through the end of the study (January 31, 2011), the combined percentages of fish moving into and out of the tower were higher (25 to 70%) than during previous months of the study. Schooling behavior was most distinct in the spring. Schooling events were present in 30 to 96% of the fish events during that period, with a peak on May 19. Schooling events were also present in the summer, but at lower numbers. With the exception of some schooling in mid-December, few to no schooling events were observed in the fall and winter months. Diel distributions for schooling fish during spring and fall months indicate schooling was concentrated during daylight hours and no schooling was observed at night. However, in December, schooling occurred at night, after midnight, and during daylight hours. Predator activity, most likely bull trout or rainbow trout according to a USACE biologist, was observed during late spring, when fish abundance index and schooling were highest for the year, and again in the fall months when fish events increased from a summer low. No predator activity was observed in the summer, and little activity occurred during the winter months

A qualitative evaluation of Southwark Council’s public health response to mitigating the mental health impact of the 2017 London bridge and borough market terror attack

Background Over recent years there have been several major terror attacks in cities across Europe. These attacks result in deaths, physical injuries, and pose long-term threats to mental health and wellbeing of large populations. Although psychologists have completed important work on mental health responses to disaster exposure including terrorist attacks, the mental health impacts of such attacks have been comparatively less examined in academic literature than the acute health response to physical injuries. This paper reflects on Southwark Council’s pioneering public mental health response to the June 2017 terror attack at London Bridge and Borough Market. It aims to explore perceptions of the mental health impact of the incident by those living and working in the borough. Methods A rapid qualitative evaluation informed by the logic underpinning Southwark Council’s response was conducted. Seven formative interviews were undertaken with individuals involved in the response planning and/or delivery, enabling the evaluation team to establish the response’s theoretical basis. Subsequently, nineteen semi-structured interviews with consenting Council employees, residents, business owners, and workers from the Borough were conducted to understand perceived mental health impacts of the attack and the success of the Council response. Thematic analysis of transcribed interviews was undertaken to evaluate the extent to which the response was implemented successfully. Results Participants reported feeling the attack had a wide-reaching negative impact on the mental health of residents, those working in the borough and visitors who witnessed the attack. Delivering the response was a challenge and response visibility within the community was limited. Participants suggested a comprehensive systematic approach to health needs assessment informed by knowledge and relationships of key Council workers and community stakeholders is imperative when responding to terrorist incidents. Improved communication and working relationships between statutory organisations and community stakeholders would ensure community groups are better supported. Prioritising mental health needs of terror attack responders to mitigate persisting negative impacts was highlighted. Conclusions This article highlights a potential public health approach and need for developing robust practical guidance in the aftermath of terror attacks. This approach has already influenced the response to the Christchurch mosque shooting in 2019.Guy’s and St Thomas’ Charity

Under pressure: Response urgency modulates striatal and insula activity during decision-making under risk

When deciding whether to bet in situations that involve potential monetary loss or gain (mixed gambles), a subjective sense of pressure can influence the evaluation of the expected utility associated with each choice option. Here, we explored how gambling decisions, their psychophysiological and neural counterparts are modulated by an induced sense of urgency to respond. Urgency influenced decision times and evoked heart rate responses, interacting with the expected value of each gamble. Using functional MRI, we observed that this interaction was associated with changes in the activity of the striatum, a critical region for both reward and choice selection, and within the insula, a region implicated as the substrate of affective feelings arising from interoceptive signals which influence motivational behavior. Our findings bridge current psychophysiological and neurobiological models of value representation and action-programming, identifying the striatum and insular cortex as the key substrates of decision-making under risk and urgency

Assessment of the Mobilizable Vector Plasmids pSUP202 and pSUP404.2 as Genetic Tools for the Predatory Bacterium Bdellovibrio bacteriovorus

Bdellovibrio and like organisms (BALOs) form the group of predatory bacteria which require Gram-negative bacteria as prey. Genetic studies with Bdellovibrio bacteriovorus can be performed with vectors which are introduced into the predator via conjugation. The usefulness of the two vectors pSUP202 and pSUP404.2 as genetic tools were assessed. Both vectors were transferable into B. bacteriovorus by conjugative matings with an Escherichia coli K12 strain as donor. The transfer frequency was higher for vector pSUP404.2 (approx. 10−1–10−4) as for pSUP202 (approx. 10−5–10−6). Vector pSUP202 with a pMB1 origin is unstable in the predatory bacterium, whereas pSUP404.2 is stably maintained in the absence of selective antibiotics. pSUP404.2 harbors two plasmid replicons, the p15A ori and the RSF1010 replication region The copy number of pSUP404.2 was determined by quantitative PCR in B. bacteriovorus and averages seven copies per genome. pSUP404.2 harbors two resistance genes (chloramphenicol and kanamycin) which can be used for cloning either by selection for transconjugants or by insertional inactivation

A new growth chart for preterm babies: Babson and Benda's chart updated with recent data and a new format

BACKGROUND: The Babson and Benda 1976 "fetal-infant growth graph" for preterm infants is commonly used in neonatal intensive care. Its limits include the small sample size which provides low confidence in the extremes of the data, the 26 weeks start and the 500 gram graph increments. The purpose of this study was to develop an updated growth chart beginning at 22 weeks based on a meta-analysis of published reference studies. METHODS: The literature was searched from 1980 to 2002 for more recent data to complete the pre and post term sections of the chart. Data were selected from population studies with large sample sizes. Comparisons were made between the new chart and the Babson and Benda graph. To validate the growth chart the growth results from the National Institute of Child Health and Human Development Neonatal Research Network (NICHD) were superimposed on the new chart. RESULTS: The new data produced curves that generally followed patterns similar to the old growth graph. Mean differences between the curves of the two charts reached statistical significance after term. Babson's 10(th )percentiles fell between the new data percentiles: the 5th to 17th for weight, the 5th and 15th for head circumference, and the 6th and 16th for length. The growth patterns of the NICHD infants deviated away from the curves of the chart in the first weeks after birth. When the infants reached an average weight of 2 kilograms, those with a birthweight in the range of 700 to 1000 grams had achieved greater than the 10(th )percentile on average for head growth, but remained below the 3(rd )percentile for weight and length. CONCLUSION: The updated growth chart allows a comparison of an infant's growth first with the fetus as early as 22 weeks and then with the term infant to 10 weeks. Comparison of the size of the NICHD infants at a weight of 2 kilograms provides evidence that on average preterm infants are growth retarded with respect to weight and length while their head size has caught up to birth percentiles. As with all meta-analyses, the validity of this growth chart is limited by the heterogeneity of the data sources. Further validation is needed to illustrate the growth patterns of preterm infants to older ages

Cord blood calcium, phosphate, magnesium, and alkaline phosphatase gestational age-specific reference intervals for preterm infants

<p>Abstract</p> <p>Background</p> <p>The objective was to determine the influence of gestational age, maternal, and neonatal variables on reference intervals for cord blood bone minerals (calcium, phosphate, magnesium) and related laboratory tests (alkaline phosphatase, and albumin-adjusted calcium), and to develop gestational age specific reference intervals based on infants without influential pathological conditions.</p> <p>Methods</p> <p>Cross-sectional study. 702 babies were identified as candidates for this study in a regional referral neonatal unit. After exclusions (for anomalies, asphyxia, maternal magnesium sulfate administration, and death), relationships were examined between cord blood serum laboratory analytes (calcium, phosphate, magnesium, alkaline phosphatase, and albumin-adjusted calcium) with gestation age and also with maternal and neonatal variables using multiple linear regression. Infants with influential pathological conditions were omitted from the development of gestational age specific reference intervals for the following categories: 23-27, 28-31, 32-34, 35-36 and > 36 weeks.</p> <p>Results</p> <p>Among the 506 preterm and 54 terms infants included in the sample. Phosphate, magnesium, and alkaline phosphatase in cord blood serum decreased with gestational age, calcium increased with gestational age. Those who were triplets, small for gestational age, and those whose mother had pregnancy-induced hypertension were influential for most of the analytes. The reference ranges for the preterm infants ≥ 36 weeks were: phosphate 1.5 to 2.6 mmol/L (4.5 to 8.0 mg/dL), calcium: 2.1 to 3.1 mmol/L (8.3 to 12.4 mg/dL); albumin-adjusted calcium: 2.3 to 3.2 mmol/L (9.1 to 12.9 mg/dL); magnesium 0.6 to 1.0 mmol/L (1.4 to 2.3 mg/dL), and alkaline phosphatase 60 to 301 units/L.</p> <p>Conclusions</p> <p>These data suggest that gestational age, as well as potentially pathogenic maternal and neonatal variables should be considered in the development of reference intervals for preterm infants.</p

Adenoviral-mediated correction of methylmalonyl-CoA mutase deficiency in murine fibroblasts and human hepatocytes

<p>Abstract</p> <p>Background</p> <p>Methylmalonic acidemia (MMA), a common organic aciduria, is caused by deficiency of the mitochondrial localized, 5'deoxyadenosylcobalamin dependent enzyme, methylmalonyl-CoA mutase (MUT). Liver transplantation in the absence of gross hepatic dysfunction provides supportive therapy and metabolic stability in severely affected patients, which invites the concept of using cell and gene delivery as future treatments for this condition.</p> <p>Methods</p> <p>To assess the effectiveness of gene delivery to restore the defective metabolism in this disorder, adenoviral correction experiments were performed using murine <it>Mut </it>embryonic fibroblasts and primary human methylmalonyl-CoA mutase deficient hepatocytes derived from a patient who harbored two early truncating mutations, E224X and R228X, in the <it>MUT </it>gene. Enzymatic and expression studies were used to assess the extent of functional correction.</p> <p>Results</p> <p>Primary hepatocytes, isolated from the native liver after removal subsequent to a combined liver-kidney transplantation procedure, or <it>Mut </it>murine fibroblasts were infected with a second generation recombinant adenoviral vector that expressed the murine methylmalonyl-CoA mutase as well as eGFP from distinct promoters. After transduction, [1-¹⁴C] propionate macromolecular incorporation studies and Western analysis demonstrated complete correction of the enzymatic defect in both cell types. Viral reconstitution of enzymatic expression in the human methylmalonyl-CoA mutase deficient hepatocytes exceeded that seen in fibroblasts or control hepatocytes.</p> <p>Conclusion</p> <p>These experiments provide proof of principle for viral correction in methylmalonic acidemia and suggest that hepatocyte-directed gene delivery will be an effective therapeutic treatment strategy in both murine models and in human patients. Primary hepatocytes from a liver that was unsuitable for transplantation provided an important resource for these studies.</p

The Evolution of Bat Vestibular Systems in the Face of Potential Antagonistic Selection Pressures for Flight and Echolocation

PMCID: PMC3634842This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited