Equipping for risk: Lessons learnt from the UK shale-gas experience on assessing environmental risks for the future geoenergy use of the deep subsurface.

findings are presented from an investigation to improve understanding of the environmental risks associated with developing an unconventional-hydrocarbons industry in the UK. The EQUIPT4RISK project, funded by UK Research Councils, focused on investigations around Preston New Road (PNR), Fylde, Lancashire, and Kirby Misperton Site A (KMA), North Yorkshire, where operator licences to explore for shale gas by hydraulic fracturing (HF) were issued in 2016, although exploration only took place at PNR. EQUIPT4RISK considered atmospheric (greenhouse gases, air quality), water (groundwater quality) and solid-earth (seismicity) compartments to characterise and model local conditions and environmental responses to HF activities. Risk assessment was based on the source-pathway-receptor approach. Baseline monitoring of air around the two sites characterised the variability with meteorological conditions, and isotopic signatures were able to discriminate biogenic methane (cattle) from thermogenic (natural-gas) sources. Monitoring of a post-HF nitrogen-lift (well-cleaning) operation at PNR detected the release of atmospheric emissions of methane (4.2 ± 1.4 t CH4). Groundwater monitoring around KMA identified high baseline methane concentrations and detected ethane and propane at some locations. Dissolved methane was inferred from stable-isotopic evidence as overwhelmingly of biogenic origin. Groundwater-quality monitoring around PNR found no evidence of HF-induced impacts. Two approaches for modelling induced seismicity and associated seismic risk were developed using observations of seismicity and operational parameters from PNR in 2018 and 2019. Novel methodologies developed for monitoring include use of machine learning to identify fugitive atmospheric methane, Bayesian statistics to assess changes to groundwater quality, a seismicity forecasting model seeded by the HF-fluid injection rate and high-resolution monitoring of soil-gas methane. The project developed a risk-assessment framework, aligned with ISO 31000 risk-management principles, to assess the theoretical combined and cumulative environmental risks from operations over time. This demonstrated the spatial and temporal evolution of risk profiles: seismic and atmospheric impacts from the shale-gas operations are modelled to be localised and short-lived, while risk to groundwater quality is longer-term.

Availability of global health opportunities in North American Paediatric Orthopaedic Fellowship Programmes.

PURPOSE: Orthopaedic residents are increasingly seeking international health electives (IHEs) during training, many of which involve providing paediatric orthopaedic care. However, little is known about the availability of IHEs during orthopaedic fellowship training. Our study sought to assess the global health opportunities available to North American paediatric orthopaedic fellows. METHODS: We conducted an online, REDCap-based survey of paediatric orthopaedic fellowship programme directors (PDs) in the United States and Canada. The survey link was sent by the Pediatric Orthopaedic Society of North America (POSNA) Evidence-Based Medicine Committee to all POSNA-approved paediatric orthopaedic fellowship PDs. Follow-up reminder emails were delivered at set time intervals. RESULTS: The overall response rate was 55% (26/47). Only three of 26 responding programmes (11.5%) offered a structured global health programme but 42.3% of programmes (11/26) reported fellow IHE participation within the last ten years. In all, 91% of PDs reported that fellows were extremely satisfied with their IHE, and 91% agreed that IHEs are valuable for trainees. Perceived barriers to fellow participation in IHEs included lack of funding, lack of established partner sites, lack of interest among fellows and concerns related to time away compromising clinical/call coverage. In all, 65.4% of PDs agree that IHE participation during training plays a major role in shaping fellows' future volunteer activities. CONCLUSION: There are limited global health opportunities among North American paediatric orthopaedic fellowship programmes, with only 11.5% offering a structured global health programme. Greater efforts to establish sustainable funding and international partnerships may increase opportunities for IHEs during paediatric orthopaedic fellowship training. LEVEL OF EVIDENCE: Level II.

Diabrotica flight in time and space as monitored with a high capacity trap placed in Zea mays fields of Illinois, USA.

Monitoring of insects by trapping is one of the prime tools of field entomologists. The leaf beetle Diabrotica virgifera virgifera LeConte (Coleoptera:Chrysomelidae) is no exception. Since its numbers (and consequently its mass) in a field population can be enormous, tools must be adapted to hold this many insects for later counting, sexing, biomass determinations, and additional investigations to follow. Since counting the high numbers during the flight peak may not be feasible at all, weighing and extrapolating to numbers by the correlation factor 1g = 160 to 170 beetles of mixed female and male sex was the method of choice. Around the perimeter of a hybrid maize (Z. mays) field of 0.6 ha, 16 high capacity traps were established at elevations ranging from 0 to 2 m above ground level. Optimal trapping is possible between 0.5 to 2m which field entomologists intuitively knew from experience and tradition. Below and above that level, the number of beetles is not zero but significantly below the optimum. High capacity traps can be left in the field with one loading of lure for four to seven days. Lures were the D. v. virgifera beetle sex pheromone 8-methyl-decane-2-ol propanoate (0.1 mg, collecting mainly male beetles) and the plant kairomone 4-methoxy-trans-cinnamaldehyde (10 mg, collecting both male and female beetles). The specific kairomone action (being much weaker than the pheromone) can be increased by simultaneously offering a feeding arrestant powder (e.g. prepared from ground Cucurbita texana or C. okeechobeensis) which keeps the beetles attracted by the kairomone lure close to the trapping site. There they can be immobilized and knocked down by a tiny source of carbaryl or dichlorvos fixed inside or at the surface of a plastic pellet or sheet. The high capacity traps are commercially available and can be reused for several flight seasons.

A promising biotechnical approach to pest management of Diabrotica virgifera virgifera in Illinois maize fields under kairomonal shielding with the new MSD technique.

Environmentally compatible and sustainable plant protection requires novel approaches to pest management characterized by minimal emphasis on toxicants. Classical toxicants traditionally dominated economic entomology for half a century. But worldwide problems with environmental pollution and with increasing resistance levels in all major pesticide classes and in many key insect species including Diabrotica virgifera virgifera (D.v.v.) strongly advocate a rethinking and a change in management paradigms used. Soft, minimally invasive, biological, biotechnical and cultural approaches should replace hard pesticides which are in favor up to now. Fortunately, pheromones, kairomones, plant attractants, better traps, new plant varieties and cultural methods like crop rotation, in short more sophisticated methods are now available as pressure for finding and exploring novel strategies increases. Facing this situation, a new biotechnical approach of population reduction of D.v.v., called "MSD" technique, is introduced. MSD is characterized as an approach combining mass trapping, shielding and deflecting of adult insects along an invisible odor barrier of synthetic kairomone which diminishes the flux of insects across a high capacity trap line baited with kairomone, thus reducing both the population fluctuation and number and its reproductive success within the shielded area. In the case of D.v.v. in Zea mays fields, effects realized by the MSD technique have been measured simultaneously by a number of independent criteria during the summers of 2003 and 2004 at 2 different locations in Illinois maize fields of up to one half hectare size. Results observed are statistically significant and cannot be explained by mass trapping alone. There is also an additional shielding and deflection, in short "diversion" effect whose basic sensory and behavioral mechanisms call for future exploration.

A monitoring trap for Diabrotica virgifera virgifera and D. barberi adults lured with a poisoned cucurbitacin bait.

A trap is described that uses as a simple natural ingredient a cucurbitacin mixture to capture and carbaryl insecticide to kill northern and western corn rootworm beetles (Diabrotica barberi Smith and Lawrence, and Diabrotica virgifera virgifera LeConte (D.v.v.), respectively). The trap is consistent in numbers of beetles captured per trap per day, and it should be useful in integrated pest management programs to monitor the population density of rootworm beetles in corn fields. Captures between 1981 and 1983 in Illinois showed that western corn rootworm adults that disperse into first-year cornfields were predominantly females. These traps are being utilized today (2003) by many researchers in the New World desiring to know more about rootworm beetle movement and activities. Entomologists in the Old World confronted with the alien invasive pest D.v.v. should profit likewise from increased knowledge and availability of these traps for Diabrotica management.

Diabrotica egg separation from soil: an efficient and fast procedure for monitoring egg stages of corn rootworm populations.

Maize growers repeatedly are confronted with the need to make predictions and decisions about which of their fields are likely to develop corn rootworm populations above the economic threshold and are in need of treatment. One of the best parameters that can help as a basis for these decisions are corn rootworm egg counts in the soil. The Western corn rootworms have one generation each year. Females oviposit eggs in the soil in corn fields from late July through early September. These eggs overwinter in the soil and almost all hatch the following June. An apparatus is described that utilizes water, MgSO4 solution and differing screen sieves for extracting the rootworm eggs from the soil samples collected from the field after deciding on an acceptable sampling procedure. Samples may be a composite of subsamples or a number of individual samples taken at various locations in a field. The Illinois machine and the final separation of eggs, using flotation in 2 molar MgSO4 solution, are highly efficient, and recoveries of 97% of rootworm eggs manually placed in samples of soil have repeatedly been achieved. Thus, this would be a useful tool in integrated pest management programs that monitor the density of corn rootworm eggs in corn fields.

Identification of an apolipoprotein(e) variant associated with type III hyperlipoproteinaemia in an indigenous Australian.

As a result of testing for lipid and apolipoprotein(e) (apo E) phenotype status of an indigenous Australian community, an apo E variant associated with type III hyperlipoproteinaemia has been identified. Apo E phenotype was determined by analysis of VLDL by isoelectric focusing, and genotype on DNA amplified by polymerase chain reaction, using two different restriction enzyme isotyping assays. Phenotypes and genotypes were discordant in samples from two subjects and an abnormal-sized restriction fragment was also observed in their genotyping gel patterns. DNA sequencing studies revealed this was due to a single nucleotide deletion, 3817delC, at amino acid 136 on apo E. This resulted in a new reading frame and the premature termination of the apo E protein due to a stop codon (TGA) at nucleotide 4105. The variant apo E null allele showed a recessive mode of inheritance and, in combination with the E2 allele, resulted in the type III hyperlipoproteinaemic phenotype but when inherited with the E4 allele had no marked effect on plasma lipids.

Familial Paget's disease of bone: nonlinkage to the PDB1 and PDB2 loci on chromosomes 6p and 18q in a large pedigree.

Paget's disease of bone is a common condition characterized by bone pain, deformity, pathological fracture, and an increased incidence of osteosarcoma. Genetic factors play a role in the pathogenesis of Paget's disease but the molecular basis remains largely unknown. Susceptibility loci for Paget's disease of bone have been mapped to chromosome 6p21.3 (PDB1) and 18q21.1-q22 (PDB2) in different pedigrees. We have identified a large pedigree of over 250 individuals with 49 informative individuals affected with Paget's disease of bone; 31 of whom are available for genotypic analysis. The disease is inherited as an autosomal dominant trait in the pedigree with high penetrance by the sixth decade. Linkage analysis has been performed with markers at PDB1; these data show significant exclusion of linkage with log10 of the odds ratio (LOD) scores < -2 in this region. Linkage analysis of microsatellite markers from the PDB2 region has excluded linkage with this region, with a 30 cM exclusion region (LOD score < -2.0) centered on D18S42. These data confirm the genetic heterogeneity of Paget's disease of bone. Our hypothesis is that a novel susceptibility gene relevant to the pathogenesis of Paget's disease of bone lies elsewhere in the genome in the affected members of this pedigree and will be identified using a microsatellite genomewide scan followed by positional cloning.

Synthesis and hybridization property of an oligonucleotide containing a 3-thioformacetal linked pentathymidylate.

A pentathymidylate fully substituted with 3'-thioformacetal intemucleotidic linkages was synthesized and subsequently incorporated into an oligonucleotide (ON) 15mer. Tm analysis was performed on the resulting ON hybridized with its complementary RNA. This duplex demonstrated slightly improved binding affinity relative to the control phosphate diester ON/RNA hybrid.

Apolipoprotein E polymorphism in indigenous Australians: allelic frequencies and relationship with dyslipidaemia.

OBJECTIVES: To determine the apolipoprotein E (apoE) allelic frequencies and the effect of apoE genotype on lipid concentrations in indigenous Australian subjects. DESIGN: Cross-sectional study. SUBJECTS AND SETTING: 155 indigenous Australians (92 women and 63 men) of mean (+/- standard deviation) age 45 +/- 17 years (SD +/- 50) were recruited without regard to history of atherosclerotic disease, in collaboration with community-based health centres in five indigenous communities in south-east Queensland. For comparison, 113 subjects of European descent and similar age distribution from the Brisbane and Gold Coast regions were also studied. MAIN OUTCOME MEASURES: ApoE allelic frequency; apoE genotype; sex; age; diabetes status; body mass index; history of atherosclerotic vascular disease; and concentrations of total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol. RESULTS: The frequency of the apoE4 allele was found to be significantly higher in the indigenous subjects than in the subjects of European descent (P < 0.001). Among indigenous subjects, those with the apoE4 allele tended to have higher triglyceride concentrations and had significantly lower HDL-cholesterol concentrations than those with the apoE3/3 and 3/2 genotypes. CONCLUSIONS: ApoE allelic frequency is likely to be one of the cluster of factors contributing to the high cardiovascular mortality of indigenous Australians.

Plasma homocysteine levels in indigenous Australians.

OBJECTIVES: To determine plasma homocysteine levels in indigenous Australians living in urban areas, and the relationship of these levels with other risk factors in this population. DESIGN: Cross-sectional study. SUBJECTS AND SETTING: 365 urban indigenous Australian subjects, 153 men and 212 women, mean (SE) age 42 (1) years, ascertained without regard to history of atherosclerotic disease, in collaboration with community-based health centres in five indigenous communities in south-east Queensland, 1997-1998. MAIN OUTCOME MEASURES: Plasma homocysteine levels, age, sex, smoking history, metformin therapy, history of atherosclerotic vascular disease, serum creatinine level, red cell folate and serum vitamin B12 levels. RESULTS: 89 subjects (24%) had plasma homocysteine levels 15 mumol/L or above. Homocysteine levels were higher in men than in women (men: 14.4 mumol/L; 95% confidence interval [CI], 13.6-15.2; women: 11.9 mumol/L; 95% CI, 11.4-12.5) (P < 0.001); correlated with age (P < 0.001); higher in current smokers (P = 0.02); higher in subjects taking metformin therapy (P = 0.007); and higher in subjects with a history of atherosclerotic vascular disease (P < 0.001). Homocysteine levels were also correlated with serum levels of creatinine (P < 0.001), red cell folate (P < 0.001), and vitamin B12 (P < 0.001). CONCLUSIONS: These data indicate that the high plasma levels of homocysteine of Australian indigenous subjects are associated with a history of vascular disease, and correlated with, among other things, smoking, and folate and vitamin B12 nutritional deficiency. These are potentially reversible risk factors, and our data suggest that focusing public health initiatives on these issues may reduce the high prevalence of cardiovascular disease in the Australian indigenous population.

The relative risks of hyperglycaemia, obesity and dyslipidaemia in the relatives of patients with Type II diabetes mellitus.

Type II (non-insulin-dependent) diabetes mellitus has a substantial genetic component; however, its molecular basis remains largely unknown. The mode of inheritance is likely to be polygenic, with penetrance influenced by environmental factors. Although the familial aggregation of Type II diabetes is acknowledged, there is little data concerning the prevalence of diabetes in the relatives of subjects with diabetes in comparison with the general population, and our objective was to address this question in the defined geographic region of Oxfordshire, England. We studied 139 first degree relatives of 90 probands with Type II diabetes who attended routine diabetes clinics in Oxfordshire and documented the fasting plasma glucose, triglyceride and HDL-cholesterol concentrations and BMI of these subjects. The probands were selected without regard to family history of diabetes. The control population data were derived from two large-scale Oxford community studies which documented the prevalences of known and newly diagnosed diabetes. The prevalences of newly diagnosed and known diabetes were calculated for each group. The mean BMI, and concentrations of fasting glucose, triglyceride and HDL-cholesterol were compared and prevalence ratios for obesity (defined as BMI > 30 kg/m2), hyperglycaemia (defined as fasting plasma glucose > or = 6.1 mmol/l), and dyslipidaemia (defined as triglyceride > 2.0 mmol/l, HDL < 1.0 mmol/l) were calculated. There was a fourfold higher prevalence of hyperglycaemia in the first degree relatives of subjects with Type II diabetes compared with the control population: the prevalence ratio after adjustment for age, sex and BMI was 4.32 (95 % confidence interval 2.29-8.17). The relatives had a considerably higher fasting plasma glucose concentration than the control population (5.18+/-0.67 mmol/l (mean +/- 1 SD) vs 4.76+/-1.59 mmol/l, p = 0.0001), and this difference remained statistically significant after adjustment for age, sex and obesity. The relatives were significantly more obese, had higher fasting plasma insulin concentrations and had lower HDL-cholesterol concentrations. In conclusion, there is a strong familial aggregation of hyperglycaemia and obesity in the relatives of subjects with Type II diabetes and these subjects have higher fasting plasma insulin concentrations and lower HDL-cholesterol than the general population. These data indicate the particular relevance of screening the first degree relatives of subjects with Type II diabetes, as intervention strategies which aim to improve the metabolic profile are indicated for a large proportion of these subjects.

Novel susceptibility gene for late-onset NIDDM is localized to human chromosome 12q.

NIDDM has a substantial genetic component, but the nature of the genetic susceptibility is largely unknown. Maturity-onset diabetes of the young (MODY) is a genetically heterogeneous monogenic form of NIDDM characterized by an early age of onset and autosomal dominant inheritance, and linkage studies have identified genes that are mutated in different MODY pedigrees on chromosome 20 (MODY1 locus, hepatocyte nuclear factor-4alpha [HNF-4alpha] gene), chromosome 7 (MODY2 locus, glucokinase gene), and chromosome 12 (MODY3 locus, HNF-1alpha gene). We studied an extended pedigree in which multiple members are affected by late-onset NIDDM associated with insulin resistance and performed linkage analysis with four microsatellite markers in the MODY3 region of chromosome 12q. We found significant evidence for linkage between NIDDM and the MODY3 locus (logarithm of odds score 3.65 at theta = 0.008 telomeric to marker D12S321), but sequencing of the 10 exons and promoter of HNF-1alpha did not identify any causative mutation in this gene. Our results indicate that the region of chromosome 12q close to MODY3 harbors a novel susceptibility gene or genes for NIDDM.

The relationship between the insulin resistance syndrome and insulin sensitivity in the first-degree relatives of subjects with non-insulin dependent diabetes mellitus.

Non-insulin dependent diabetes mellitus (NIDDM) has a substantial genetic component. Impaired insulin secretion, insulin insensitivity in muscle and adipose tissue, and elevated hepatic glucose production are the major pathophysiological features of NIDDM. Insulin insensitivity is also a feature of the insulin resistance syndrome, which describes the epidemiological association of glucose intolerance, upper body obesity, hyperinsulinaemia, hypertension, increased triglyceride levels and decreased high-density-lipoprotein (HDL)-cholesterol concentrations. Insulin insensitivity has been found to be a familial trait, and this raises the hypothesis that the insulin resistance syndrome may also occur as a familial trait in caucasian families in association with the development of NIDDM. The 90 first degree relatives of 50 caucasian subjects with NIDDM were studied with a continuous infusion glucose tolerance test to quantitate glucose tolerance, insulin sensitivity and beta-cell function. Body mass index (BMI), blood pressure, fasting triglyceride and HDL-cholesterol measurements were obtained, and the intercorrelations between these variables were examined. As a group the first degree relatives had a median insulin sensitivity of 65% (interquartile range 46-99%). Insulin sensitivity was univariately correlated with systolic and diastolic blood pressure, triglyceride and HDL-cholesterol. These associations were present in both the hyperglycaemic and the normoglycaemic relatives. The hyperglycaemic relatives were significantly more insulin insensitive than the normoglycaemic relatives, but this additional insulin insensitivity was not associated with significant differences in blood pressure, triglyceride or HDL-cholesterol concentrations. Our data indicate that the insulin insensitivity present in the first degree relatives of subjects with NIDDM is correlated with the cardiovascular risk factors which make up the insulin resistance syndrome, and that glycaemic status does not appear to be the major determinant of these associations. Interventions targeting obesity and insulin insensitivity in these subjects may reduce cardiovascular risk.

Type II diabetes: clinical aspects of molecular biological studies.

Type II diabetes remains a genetic nightmare. The major problem is identifying suitable pedigrees, sib-pairs, and populations for study. Segregation analysis data suggest that type II diabetes is likely to be polygenic, although one or more major genes could also be involved. This and the high prevalence of diabetes affect the strategies for searching for genetic mutations. Linkage analysis in classical type II diabetes pedigrees is unlikely to be successful. In addition, affected sib-pair analysis is limited because both parents are often affected, leading to bilineal inheritance. Sib-pairs with both parents alive are unusual, so identity by descent analysis is rarely feasible. Strategies to reduce bilineal inheritance by identifying sib-pairs with one known nondiabetic parent or with the second sibling having mild subclinical diabetes may be worthwhile. Identification of individuals or pedigrees with an unusual phenotype that suggests a single gene disorder, such as maturity-onset diabetes of the young, will continue to be important, for this allows linkage analysis with markers near candidate genes and exclusion mapping of chromosomal regions using highly polymorphic markers. Population association studies with candidate genes can detect mutations that have a minor role in the majority proportion of diabetic subjects, but large numbers are required and great care must be taken to exclude ethnic group differences between the diabetic and normoglycemic populations. The study of small inbred communities might be helpful because they may have fewer diabetogenic genes than outbred populations, and this would increase the power of sib-pair and population association studies. Direct screening for mutations in candidate genes (with single-strand conformation polymorphism or heteroduplex screening or with direct sequencing) in patients with the appropriate pathophysiological abnormality can be a successful strategy. The identification of well-defined diabetic pedigrees, sib-pairs, and suitable matched diabetic and nondiabetic populations will be key to the discovery of the genes for diabetes.

Fluconazole and testosterone: in vivo and in vitro studies.

Fluconazole (UK-49,858), a novel bis-triazole antifungal agent, was given orally to groups of 10 male volunteers at doses of 25 and 50 mg/day for 28 days. Blood samples for testosterone estimation were taken from these and from a placebo group at several time points on days 1, 14, and 28 of the study, and the assay results demonstrated that the compound had no significant effect on circulating testosterone levels. Similarly, in studies with rat Leydig cells in vitro, fluconazole at concentrations up to 10 micrograms/ml was found to be only a weak inhibitor of testosterone production, whereas ketoconazole caused more than 50% inhibition at 0.1 microgram/ml. It is concluded that fluconazole, in contrast to ketoconazole, has little effect on the biosynthesis of testosterone by mammalian cells.

In vitro effects of fluconazole (UK-49,858) and ketoconazole on mouse lymphocyte proliferation and on Candida blastospore destruction by human polymorphonuclear leukocytes.

The effects of fluconazole, a novel antifungal compound with a bis-triazole structure, were compared with those of ketoconazole on mitogen-induced DNA synthesis by cultured mouse lymphocytes, and on the destruction of fungal (Candida albicans) blastospores by human polymorphonuclear leukocytes. Fluconazole had little or no effect on concanavalin A- or lipopolysaccharide-induced lymphocyte proliferation at concentrations at which ketoconazole caused marked inhibition of the response to both these mitogens. Similarly, fungal cell killing by polymorphonuclear leukocytes was substantially depressed by ketoconazole but was unaffected by fluconazole. In addition, therefore, to having excellent in vivo antifungal activity, fluconazole, unlike ketoconazole, has been shown to have no inhibitory effect in two in vitro assays of immune function.

The origin of fibrin breakdown products and the interpretation of their appearance in the circulation.

It has been shown that the incubation of human plasma with urokinase at a concentration sufficient to cause rapid lysis of the clots formed on the addition of thrombin does not give rise to the production of measurable concentrations of non-clottable fibrinogen breakdown products. Also, breakdown products could not be detected in the course of experiments in vivo when urokinase was administered to monkeys and only in very low concentrations when a fibrinolytic state was induced by exercise in three healthy human volunteers. In contrast, high concentrations of breakdown products were found after thrombin infusion into monkeys. It is concluded that circulating fibrinogen is not readily broken down into non-clottable products by the fibrinolytic enzymes, and that normal animals and healthy human subjects do not have substantial deposits of fibrin that are available for breakdown during a fibrinolytic episode. The presence of breakdown products in the circulation is therefore likely to be indicative of the fibrinolytic response to an initial coagulation event.