Exploring how fire spread mode shapes the composition of pyrogenic carbon from burning forest litter fuels in a combustion wind tunnel.

In this study, solid state 13C nuclear magnetic resonance (NMR) spectroscopy was used to explore the carbon-containing functional groups present in pyrogenic carbon (PyC) produced during different fire spread modes to forest litter fuels from a dry sclerophyll eucalypt forest burnt in a combustion wind tunnel. A replicated experimental study was performed using three different fire spread modes: heading fires (i.e. fires which spread with the wind), flanking fires (i.e. fires which spread perpendicular to the wind) and backing fires (i.e. fires which spread against the wind). In addition to 13C NMR measurements of PyC, detailed fire behaviour measurements were recorded during experiments. Experiments showed that heading fires produced significantly more aryl carbon in ash samples than flanking fires. All other experimental comparisons for burnt fuel samples involving different fire spread modes were statistically insignificant. Principal component analysis (PCA) was used to explore the relationship between 13C NMR functional groups and fire behaviour observations. Results from PCA indicate that maximising the residence time of high temperature combustion and the combustion factor (i.e. the fraction of pre-fire biomass consumed by fire) could be a method for increasing the amount of aryl carbon in PyC. Maximising the amount of aryl carbon could be beneficial for the overall PyC balance from fire, since more recalcitrant carbon (e.g. carbon with a higher aryl carbon content) that is not emitted to the atmosphere has been shown to have longer residence times in environmental media such as soils or sediments.

The effect of fire affected Pinus radiata litter and char addition on soil nitrogen cycling.

In pine forest litters, decomposition rate is directly affected by the pathway the needle followed to the ground, whether that was via programmed apoptosis and abscission or via stress induced loss through branch damage or tree death. Stress induced losses may occur due to fire damage, which leads to a post-fire litter layer composed of non-senescent debris that fell during or after the event. This study investigates decomposition and nitrogen cycling in soils amended with two litters from Pinus radiata plantations that had different recent fire histories. Litters were incubated in the presence or absence of field collected char for up to 94 days. These soil treatments were analysed for microbial activity (soil respiration) and N pools (microbial, mineral, and potentially mineralisable). Soil and litter treatments were additionally incubated in the presence of ammonium nitrate solution to determine N absorption potential of the litters. Respiration was greatest in soils that received fire affected (FA) litter regardless of the presence or absence of char. Nitrogen pools were largely similar between the control (no litter) treatment and not fire affected (NFA) litter treatments. Measured N pools were exceedingly low (92% of samples <2 µg-N g soil-1 where detected) or not detectable (37% of samples below detection limits) in all FA litter treatments at most times. Char appeared inert throughout and had no effects on microbial activity or nitrogen cycling. This study indicates that fire affected pine litter collected four months post fire has strong N absorption properties with or without the presence of char. The presence of fire affected litter is likely to affect N availability for regeneration of forest growth.

Microbial DNA profiling by multiplex terminal restriction fragment length polymorphism for forensic comparison of soil and the influence of sample condition.

AIMS: To evaluate: (i) the impact of air-drying on bacterial, archaeal and fungal soil DNA profiles and (ii) the potential use of multiplex-terminal restriction fragment length polymorphism (M-TRFLP) as a tool for forensic comparison of soil. METHODS AND RESULTS: An M-TRFLP approach was used to profile bacterial, archaeal and fungal DNA profiles from five different soil sites. Air-drying soil significantly reduced the quantity of DNA but the number of operational taxanomic units (OTU) was unaffected. The impact of air-drying on soil DNA profiles was dependent on soil site and microbial primers. Fungal profiles were altered the least by air-drying. For prokaryotic profiles, air-drying altered the relative similarity/dissimilarity between soil sites. The M-TRFLP approach was more discriminatory compared with soil colour and single-taxa profiling, but did not significantly improve resolution between two similar soils. CONCLUSIONS: Of those tested, soil fungi were potentially the more robust target for application to soil forensic studies as they were altered less by air-drying and provided clear discrimination of soils from different sites. The M-TRFLP method demonstrated potential to achieve greater resolution, discriminating the soil sites based on both bacterial and fungal components. SIGNIFICANCE AND IMPACT OF THE STUDY: Soil DNA profiling has potential as a forensic tool, but sample condition and the appropriate selection of microbial target taxa must be considered.

Pain assessment tools: children's nurses' views.

The persistence of inadequate treatment of pain in children could be due to lack of knowledge and nurses' failure to assess and manage pain effectively. It is recognized that effective pain assessment leads to more satisfied children and families. This study explored children's nurses' views on the use of pain assessment tools in a tertiary referral centre. Almost two-thirds of nurses did not have a preference for a pain assessment tool, but nearly three-quarters of nurses surveyed agreed that the introduction of pain assessment tools would improve documentation. When nurses were asked how much time they needed for education on these tools, 83 percent wanted only two hours, although almost half stated lack of knowledge or education as the main obstacle to use of a pain assessment tool. The inconsistencies in these replies could reflect the conflicting demands between the nurses' need to increase their knowledge of pain assessment while managing a heavy workload.

Efficacy of oryzalin and associated histological changes in Cryptosporidium-infected neonatal rats.

This paper reports the anti-cryptosporidial effects of, and concomitant amelioration of the histological changes in the gut of neonatal rats with intestinal cryptosporidiosis treated with the dinitroaniline, oryzalin. The ED50 was determined to be 7 mg/kg using twice daily doses administered for 3 consecutive days. A maximum inhibition of 85.5% was achieved at 25 mg/kg and this inhibition remained constant despite increasing the oryzalin dose to 200 mg/kg. Cryptosporidiosis significantly decreased the intestinal villus/crypt (VC) ratio by approximately 50% (duodenum = 2.3, jejunum = 2.5 and ileum = 1.7) when compared to uninfected untreated controls (duodenum = 4.3, jejunum = 5.9 and ileum = 4.5). Treatment with oryzalin doubled the VC ratio in the duodenum, jejunum and ileum following doses of 5 mg, 50 mg and 200 mg/kg respectively. Oryzalin concentrations in the small intestine contents and plasma were determined, using HPLC, at 0.5, 1 and 2 h after dosing. The much greater dose required to return VC ratios to normal in the ileum (200 mg/kg) compared to the duodenum (6.25 mg/kg) appeared to reflect the decreased concentration of the drug in the distal small intestine. Concentrations of oryzalin equivalent to the in vitro IC50 were maintained for 2 h in the first half of the small intestine following a single dose of 100 mg/kg.

Expression of Giardia duodenalis beta-tubulin as a soluble protein in Escherichia coli.

The beta-tubulin gene of the parasitic protozoan Giardia duodenalis has been expressed for the first time using a novel and direct method. The protein was expressed in both soluble and insoluble forms in an Escherichia coli-based expression system. The level of expression was found to be affected by several variables including the incubation temperature, length of time for which expression was carried out, and the E. coli culture volume. The protein expression system contributed no additional amino acids to the final fusion protein and the polyhistidine fusion sequence was easily removed from the beta-tubulin protein using a specific enterokinase enzyme. The expression system also provided a means of preparing a soluble protein and purifying it by a relatively straightforward affinity chromatography method to give a very high level of protein purity. This makes the protein suitable for a number of applications for characterization including beta-tubulin antibody assays, alpha-/beta-tubulin-binding regions, and beta-tubulin folding intermediates.

A comparison of the effects of a benzimidazole and the dinitroanilines against Leishmania infantum.

Leishmania infantum promastigotes and amastigotes were axenically cultured and exposed to the known tubulin binding compounds, the dinitroanilines, trifluralin, benfluralin, pendimethalin, oryzalin and the precursor of the dinitroanilines, chloralin, as well as isomers of chloralin and trifluralin and to the benzimidazole, albendazole. Drug induced inhibition was observed using [3H]thymidine uptake compared with untreated controls. In vitro analysis demonstrated a significant difference in the activity of five of the seven dinitroanilines between both life cycle stages of L. infantum. The amastigotes were 20-times more sensitive to chloralin and its isomer than to the dinitroanilines whereas the promastigotes were similar in sensitivity to the dinitroanilines and to chloralin and its isomer. This interesting finding suggests that the dinitroaniline precursors may have different target sites in the amastigotes to those within the promastigotes. Additionally, both chloralin and its isomer, and to a lesser extent benfluralin, caused a substantial stimulation of thymidine incorporation (up to 50%) at low concentrations. Dose response analysis suggests that the dinitroanilines may have more than one mode of action against L. infantum amastigotes and promastigotes. The inhibitory effects of the dinitroanilines against L. infantum vary from previous findings using the dinitroanilines against other Leishmania spp. The 348 base pair DNA sequence coding for beta-tubulin from amino acid residues 132 to 248 was obtained for L. infantum and used to compare the in vivo efficacy of albendazole with predicted activity based on beta-tubulin sequences of known benzimidazole sensitive protozoa. The use of beta-tubulin sequence as a predictive model of benzimidazole activity is discussed with particular reference to L. infantum.

A comparison of the effects of two dinitroanilines against Cryptosporidium parvum in vitro and in vivo in neonatal mice and rats.

The effects of two dinitroanilines, oryzalin and trifluralin, were compared against Cryptosporidium parvum, in vitro using HCT-8 cells and in vivo using neonatal Swiss ARC mice and Wistar neonatal rats. In vitro, oryzalin and trifluralin exhibited IC(50) values (concentration necessary to cause a 50% inhibition) of 750 and 800 nM, respectively. A viability assay showed that neither compound produced a cytotoxic effect on the host cells at concentrations as high as 1 microM. The in vivo component of this study consisted of inoculation of neonatal mice and neonatal rats with 10(5) viable oocysts of C. parvum per animal and the subsequent treatment of this infection with trifluralin and oryzalin administered via gastric intubation. At doses of 100 mg kg(-1) body weight administered twice daily for 3 consecutive days, trifluralin had no statistically significant effect on the number of oocysts recovered from the gut of either rats or mice compared with controls, whereas at the same concentration, oryzalin caused 90 and 79% inhibition of oocysts recovered from mice and rats, respectively.

High resolution ultrasound assessment of the carotid artery: its relevance in postmenopausal women and the effects of tibolone on carotid artery ultrastructure.

OBJECTIVES: Hormone replacement therapy protects from cardiovascular disease at the menopause in part by reduction of menopausal pro-atherogenic serum lipid changes. Tibolone has beneficial effects on lipids, although serum high density lipoprotein levels decrease. This study aimed primarily to establish the effects of long-term administration of tibolone on a new surrogate marker for cardiovascular disease risk, the measurement of carotid artery intima-media thickness (CIMT) using high-resolution ultrasound. METHODS: Measurement of CIMT and assessment of carotid atherosclerotic plaques were undertaken in 31 women on tibolone and 30 voluntary controls from an ongoing open-label study of tibolone. RESULTS: The two groups were comparable, except for mean age and prevalence of current smokers. Repeatability of CIMT measurements was acceptable (CV, 10.0%). CIMT was significantly thicker in those with atherosclerotic plaques and increased systolic blood pressure. Prevalence of plaques was raised in those who had ever smoked, and those with elevated systolic blood pressure. There was no influence of tibolone on CIMT, whether plaques were present or not. CONCLUSIONS: This reliable technique demonstrates associations between CIMT and established risk factors. CIMT was significantly thicker in those with existing plaques. We did not demonstrate an effect of long-term tibolone use on either CIMT or prevalence of plaques.

Spleen sizing by ultrasound in polycythaemia and thrombocythaemia: comparison with SPECT.

Detection of non-palpable early splenic enlargement may aid diagnosis of primary polycythaemia (PP) and primary thrombocythaemia (PT). In this study linear spleen sizing by ultrasound has been compared with spleen volume estimation by single photon emission computerized tomography (SPECT) in 26 patients. Spleen length by ultrasound correlated well with SPECT volume estimation. Ultrasound spleen length was also measured in 60 normal control subjects where the upper limit of the 95% reference range was 11.6 cm. Changes in spleen length with both age and body weight were substantial and overshadowed the imperfect reproducibility of this method. Therefore, interpretation of an individual's measured spleen length should be in relation to that predicted for adults of the same age and weight, particularly at the extremes of the younger, heavier patients and also the older, lighter patients. Ultrasound spleen lengths of different patient groups (21 PP, 26 PT, 17 idiopathic erythrocytosis, 12 secondary polycythaemia, nine apparent polycythaemia) were compared both using the measured overall reference range and the differences from the values predicted for their age and weight. The comparison showed that almost all patients with PP whose spleens were not palpable had spleen lengths greater than the upper limit for the normal control group, but separation from the other patient groups was incomplete. Detection of non-palpable splenomegaly by ultrasound length should remain a 'minor' criterion amongst the 'proposed modified diagnostic criteria' of PP.

Measuring health-related quality of life outcomes in women with endometriosis--results of the Gynaecology Audit Project in Scotland.

The clinical management of endometriosis was addressed within the recent Gynaecology Audit Project in Scotland. The impact of endometriosis and its treatment on women's health-related quality of life was examined using a condition-specific measure and a general measure, the Short Form 36 health survey (SF-36). Postal questionnaires containing the health-related quality of life measures were sent to 273 women at diagnosis and six months later. The measurement properties, including the reliability, validity and responsiveness, of the measures were examined. The condition-specific questions and the SF-36 had a high level of reliability. The validity of the condition-specific scores was demonstrated by their high correlation with the SF-36 which is a well-validated measure. Furthermore, the condition-specific scores were related to clinicians' assessment of disease severity and the need for further treatment. At the six month follow-up, changes in scores conformed to expected hypotheses, demonstrating the responsiveness of both measures. As a general measure, the SF-36 appeared to reflect the effects of both the condition of interest (i.e. endometriosis) and other conditions affecting health at the time of measurement (i.e. treatment side effects). The condition-specific measure was more responsive than the SF-36 to the changes in pain symptoms which resulted from active treatment. A condition-specific questionnaire, together with a general measure such as the SF-36 health survey, can provide a reliable, valid and responsive package of measures for assessing health-related quality of life in women with endometriosis. Such measures should be used alongside clinical measures of outcome to assess the effectiveness of different treatment strategies for endometriosis. A similar approach combining general and specific instruments would be useful in medical audits of other conditions.

The incidence of transient renal medullary hyperechogenicity in neonatal ultrasound examination.

A prospective ultrasound study of the urinary tracts of 85 neonates (64 term, 21 preterm) was performed to assess the incidence of transient renal medullary hyperechogenicity (RMH) in the first week of life. None of the neonates examined had evidence of renal dysfunction. Echogenic material was observed in the renal papillae/calyces, ureter, or bladder of 33 of the 64 term babies, but in the bladder of only one pre-term infant. The distribution of the echogenicity differs from that seen in medullary hyperechogenicity due to crystal deposition, suggesting that calyceal involvement is a common feature. Follow-up ultrasound scans at 10-14 days were possible in eight of the term neonates and demonstrated complete resolution of the RMH. The aetiology of transient neonatal RMH is unclear, although it may be related to protein cast deposition in the renal tubules. RMH may rarely be associated with transient renal dysfunction, but in healthy neonates should be recognized as a normal variant.

The ultrasound appearances of neonatal renal vein thrombosis.

Renal vein thrombosis (RVT) is the most frequently occurring vascular condition in the new-born kidney. The predisposing factors include dehydration, sepsis, birth asphyxia, maternal diabetes, polycythaemia and the presence of an indwelling umbilical venous catheter. (RVT) may present clinically with a flank mass, haematuria, hypertension or renal failure. Many imaging modalities have been employed, but ultrasound is the technique most commonly used in the evaluation of neonates with suspected RVT. Thrombosis commences in the small renal veins and subsequently propagates via larger interlobar veins to the main renal vein and inferior vena cava (IVC). The ultrasound appearances depend upon the stage at which the examination is performed and extent of the thrombus. Initially, the interlobular and interlobar thrombus appears as highly echogenic streaks. These streaks commence in a peripheral, focal segment of the involved kidney and only persist for a few days. In the first week the affected kidney swells and becomes echogenic with prominent echopoor medullary pyramids. Later, the swelling increases and the kidney becomes heterogenous with loss of corticomedullary differentiation. Grey scale ultrasound readily demonstrates thrombus within the renal vein and IVC. Adrenal haemorrhage is a recognized association and may be identified ultrasonically. Colour Doppler scanning provides additional information. In the early stages of RVT, colour Doppler may demonstrate absent intrarenal and renal venous flow. Ultimately, the kidney may recover, show focal scarring or become atrophic. Thus, ultrasound provides an accessible and reliable tool in the assessment of suspected neonatal RVT.

Transcripts of Grg4, a murine groucho-related gene, are detected in adjacent tissues to other murine neurogenic gene homologues during embryonic development.

The groucho-related genes (Grg) of the mouse comprise at least four family members. In Drosophila, groucho is one of the neurogenic genes that participates in the Notch signalling pathway. The Groucho protein interacts with Hairy-related transcription factors to regulate segmentation, neurogenesis and sex determination. Thus, by analogy to the Drosophila proteins, murine Grg proteins may interact with mammalian Hairy and E(spl) homologues (Hes proteins) and take part in a signalling pathway downstream of murine Notch. We have isolated murine Grg4 cDNAs and examined Grg4 expression during embryogenesis. Transcripts of Grg4 were detected in proliferating epithelial tissues undergoing mesenchymal induction, overlapping with Grg3, Notch1 and Hes1 expression. Grg4 was also expressed in the central nervous system and somites, but in cells adjacent to Grg3-, Notch1-, and Hes1-expressing cells. This distinct pattern of expression suggests a role for Grg4 in later stages of cell differentiation than for the other mouse neurogenic gene homologues.

A new approach to the measurement of quality of life. The Patient-Generated Index.

Quality of life has been defined as "the extent to which our hopes and ambitions are matched by experience." To improve a patient's quality of life through medical care would be to "narrow the gap between a patient's hopes and expectations and what actually happens." Using the above definition as a conceptual basis, we produced a self-administered, Patient-Generated Index (PGI) of quality of life. The PGI was completed by 359 patients presenting with low back pain. The validity of the measure was assessed by correlating patients' PGI scores with a well-validated health profile, the Short-Form 36-item Health Survey (SF-36), and with their scores on a clinical back pain questionnaire. Stepwise multiple regression was then used to model the relationship between the PGI score and the SF-36. Patients' PGI scores showed a high correlation with SF-36 scales measuring pain, social functioning, and role limitations attributable to physical problems, and with the clinical questionnaire. Together with whether a person was retired or not, these health variables were able to explain 25% of the variance in PGI scores. Patient generated index scores were significantly lower in patients referred to hospital compared with those managed solely in general practice and tended to reflect the general practitioner's assessment of symptom severity. We conclude that it is possible to construct a questionnaire that quantifies the effect of a medical condition on patients' quality of life in a way that has meaning and relevance in the context of their daily lives. The PGI has considerable potential for routine use in a wide range of clinical conditions for which the measurement of outcome has hitherto proved very difficult.

Increased levels and constitutive tyrosine phosphorylation of the epidermal growth factor receptor contribute to autonomous growth of human papillomavirus type 16 immortalized human keratinocytes.

Transfection of individual normal human foreskin keratinocyte (HKc) strains with human papillomavirus type 16 (HPV16) DNA results in the establishment of immortalized cell lines (HKc/HPV16) which, like normal HKc, require epidermal growth factor (EGF) and bovine pituitary extract (BPE) for proliferation in serum-free media. However, sublines which proliferate in serum-free media in the absence of EGF and BPE can be reproducibly established from individual HKc/HPV16 lines, following selection in serum-free media lacking EGF and BPE. The growth factor-independent sublines (HKc/GFI) proliferate in the absence of EGF and BPE at the same rate and to the same extent as in medium supplemented with these growth factors, whereas the parental HKc/HPV16 lines proliferate poorly in the absence of EGF and BPE. As a first approach to understanding the molecular basis by which HKc/GFI have lost their requirement for EGF, we compared EGF uptake and EGF receptor (EGFR) numbers in normal HKc, HKc/HPV16, and HKc/GFI. HKc/GFI exhibit increased EGF uptake and increased EGFR numbers compared to HKc/HPV16. A neutralizing antibody against the extracellular domain of the EGFR dramatically inhibited clonal growth of HKc/GFI, indicating that signaling through the EGFR must be important for the ability of HKc/GFI to proliferate in the absence of EGF. In addition, while in the absence of EGF normal HKc and HKc/HPV16 exhibited no detectable EGFR tyrosine phosphorylation, the EGFRs in HKc/GFI were tyrosine phosphorylated in the absence of EGF and hyperphosphorylated in the presence of EGF. Although an anti-TGF-alpha antibody inhibited the growth of HKc/GFI, we unexpectedly found that HKc/GFI and HKc/HPV16 secreted comparable and extremely low amounts of TGF-alpha (4 to 9 pg/10(6) cells per 24 h); about 100- to 250-fold less than normal HKc (1018 pg/10(6) cells per 24 h). No other ligands for the EGFR were detected in media conditioned by normal HKc, HKc/HPV16, or HKc/GFI. Thus, while overexpression and constitutive activation of the EGFR appear to be important features of HKc/GFI, enhanced secretion of TGF-alpha or other ligands for the EGFR does not explain the proliferation of HKc/GFI in the absence of EGF and BPE.

Octreotide: a long-acting inhibitor of endogenous hormone secretion for human metabolic investigations.

Octreotide is a recently available, FDA-approved, long-acting analog of somatostatin. The efficacy and tolerability of octreotide were evaluated in a series of protocols in healthy volunteers to assess its suitability for use in clinical investigations involving short-term inhibition of endogenous hormone secretion. Prolonged (270 minutes) hyperglycemic clamps were used to assess octreotide-mediated suppression of glucose-stimulated endogenous insulin secretion. Compared with a saline-control infusion, octreotide (30 ng/kg/min) suppressed stimulated insulin (P < .0001) and C-peptide (P < .0001) concentrations to basal levels. During insulin-induced hypoglycemia (plasma glucose < 40 mg/dL), octreotide (30 ng/kg/min) effectively suppressed the secretion of glucagon (P < .05) and growth hormone (P < .0005). In islet cell clamp studies, octreotide (30 ng/kg/min) suppressed C-peptide (P < .001), glucagon (P < .01), and growth hormone concentrations to below basal (fasting) levels in all subjects. Subsequent infusion of exogenous insulin, glucagon, and growth hormone resulted in predictable and stable concentrations of each hormone during octreotide-mediated suppression of their endogenous secretion. Consistent with the long half-life of octreotide (approximately 90 minutes), the concentrations of all three hormones remained suppressed below basal levels throughout a 60-minute observation period following the termination of octreotide infusion. In separate high-dose octreotide infusion studies, octreotide (60 ng/kg/min) did not produce any apparent additional metabolic effects, but was associated with an unacceptable degree of gastrointestinal side effects.(ABSTRACT TRUNCATED AT 250 WORDS)