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Meticillin-resistant Staphylococcus aureus (MRSA) outbreaks have been reported previously in burns centres with resulting mortality and morbidity. This article describes the first human-associated outbreak in the UK caused by a strain of mupirocin-resistant (MuR) livestock-associated MRSA clonal complex 398 (LA-MRSA CC398) in an adult burns centre. The centre historically had a very low prevalence of MRSA infections.To describe the clinical and epidemiological context of how the outbreak was identified and contained using a range of infection prevention and control (IPC) measures guided by both traditional and genetic methods.A cluster of MuR-MRSA led to an outbreak investigation. Cases were detected via retrospective search and real-time laboratory surveillance. Isolates were sent continuously for whole-genome sequencing (WGS). A live timeline of cases and interventions was produced throughout the period.The outbreak consisted of 12 cases (seven males and five females) aged between 22 and 70 years. Patients were identified between May and October 2020. All patients were colonized rather than infected. The strain acquired the plasmid bearing MupA while colonizing the index case before dissemination. The index case was found to be a chicken farmer. This outbreak was eventually controlled using IPC measures, audits, and blind staff decolonization guided by insight from WGS.It was not possible to determine how the strain entered the centre, or if a staff carrier was involved. The outbreak demonstrated the potential for continued transmissions for months despite active surveillance and stringent control measures.

Introduction Despite COVID-19 being a viral illness, antibiotic use has been more prevalent. In addition, co-infection (3.5%) and secondary infection (14.3%) were relatively low in hospitalised patients with COVID-19. A major concern is the increased risk of antimicrobial resistance (AMR) due to inappropriate antibiotic consumption (1). Aim This review aims to evaluate antimicrobial consumption (excluding repurposed drugs such as remdesivir) in hospitals and determine the prevalence of COVID-19 patients who received antibiotic therapy using meta-analysis. Methods The review was conducted according to PRISMA guidelines (2). The two investigators independently developed and applied eligibility criteria to examine original articles. Studies were eligible for inclusion if they met the following criteria: (i) original research studies with a minimum sample of 50 patients; (ii) focussed on antibiotic consumption (AMC); (iii) patients with COVID-19 or consumption amid COVID-19 pandemic; (iv) any age group or gender; and (v) reported in the English language. The included articles were retrieved from MEDLINE, CINAHL, WHO COVID-19 databases, including studies published in EMBASE, Scopus, WHO-COVID, and LILACS between December 2019 to July 2021. The modified version of Newcastle-Ottawa Scale (NOS) was used to measure biases in included studies after the consensus by both authors. The random-effects model was used to estimate the pooled prevalence or proportion of AMC among hospitalized COVID-19 patients. Results A total of 34 studies conducted among hospitalised COVID-19 patients were included. The extracted studies presented AMC in defined daily doses (DDD) or frequency and percentages. Azithromycin was the most frequently prescribed antibiotic in almost all studies. The meta-analysis that examined overall AMC using data from 25 studies (17 studies from high income countries and eight from low-middle income countires) revealed 69% (95% CI:63%-74%) of hospitalized COVID-19 received at least one course of antibiotics. The sub-group analysis of studies from high income countries (HICs) revealed 59% (95% CI: 51%-66%) consumed antibiotics compared with 89% (95% CI: 82% to 94%) among hospitalised COVID-19 patients in low-middle income countries (LMICs). Conclusion This review highlights the trend of antibiotic consumption in hospitalised COVID-19 patients. A significant rise in antibiotic consumption was observed in LMICs and increased antibiotic consumption in the first few months of the COVID-19 pandemic in HIC. The review outcomes emphasised the importance of rational and judicious use of antimicrobial therapy as well as to strenghting the antimicrobial stewardship policies and activities, particularly during a global pandemic. The limitation of the review undertaken was not identified incidence of co-infection and don’t include studies on reported AMC in immunocompromised patients. References (1) Rawson TM, Ming D, Ahmad R, Moore LSP, Holmes AH. Antimicrobial use, drug-resistant infections and COVID-19. Nature reviews Microbiology. 2020;18(8):409-10. (2) Beller EM, Glasziou PP, Altman DG, Hopewell S, Bastian H, Chalmers I, et al. PRISMA for Abstracts: Reporting Systematic Reviews in Journal and Conference Abstracts. PLOS Medicine. 2013;10(4):e1001419.

Molecular hydrogen (H2), its stable isotope signature (δD), and the key combustion parameters carbon monoxide (CO), carbon dioxide (CO2), and methane (CH4) were measured from various combustion processes. H2 in the exhaust of gas and oil-fired heaters and of waste incinerator plants was generally depleted compared to ambient intake air, while CO was significantly elevated. These findings contradict the often assumed co-occurring net H2 and CO emissions in combustion processes and suggest that previous H2 emissions from combustion may have been overestimated when scaled to CO emissions. For the gas and oil-fired heater exhausts, H2 and δD generally decrease with increasing CO2, from ambient values of ~0.5 ppm and +130‰ to 0.2 ppm and −206‰, respectively. These results are interpreted as a combination of an isotopically light H2 source from fossil fuel combustion and a D/H kinetic isotope fractionation of hydrogen in the advected ambient air during its partial removal during combustion. Diesel exhaust measurements from dynamometer test stand driving cycles show elevated H2 and CO emissions during cold-start and some acceleration phases. While H2 and CO emissions from diesel vehicles are known to be significantly less than those from gasoline vehicles (on a fuel-energy base), we find that their molar H2/CO ratios (median 0.026, interpercentile range 0.12) are also significantly less compared to gasoline vehicle exhaust. Using H2/CO emission ratios, along with CO global emission inventories, we estimate global H2 emissions for 2000, 2005, and 2010. For road transportation (gasoline and diesel), we calculate 8.3 ± 2.2 Tg, 6.0 ± 1.5 Tg, and 3.8 ± 0.94 Tg, respectively, whereas the contribution from diesel vehicles is low (0.9–1.4%). Other fossil fuel emissions are believed to be negligible but H2 emissions from coal combustion are unknown. For residential (domestic) emissions, which are likely dominated by biofuel combustion, emissions for the same years are estimated at 2.7 ± 0.7 Tg, 2.8 ± 0.7 Tg, and 3.0 ± 0.8 Tg, respectively. For biomass burning H2 emissions, we derive a mole fraction ratio ΔH2/ΔCH4 (background mole fractions subtracted) of 3.6 using wildfire emission data from the literature and support these findings with our wood combustion results. When combining this ratio with CH4 emission inventories, the resulting global biomass burning H2 emissions agree well with published global H2 emissions, suggesting that CH4 emissions may be a good proxy for biomass burning H2 emissions.

Buildings account for around 30% of the developed countries greenhouse gas (GHG) emissions. Improving energy effi ciency of buildings is one of the quickest and most cost-effective ways of reducing GHG emissions. This paper outlines the results of research carried out in Australia in 2009. The broad aims of the research are to identify policy directions to aid in the uptake of sustainability practices that will help improve building performance and reduce GHG emissions. Part of the research entailed surveying building practitioners involved in residential property development to fi out their experiences with designing and constructing homes and what they perceive to be the drivers and barriers to the uptake of sustainable building practices. Respondents have seen an increase in demand for energy-effi cient ‘green’ homes, due to an increase in awareness of climate change and also the reduced utility costs associated with these homes. The most common, client-preferred, and successful features incorporated into the design and retrofi tting of homes were passive solar design, having gas connected, solar water heating and rainwater tanks. The high sunshine hours in Australia together with the availability of Government rebates and subsidies make these features fi nancially viable. The average cost premium to build a ‘green’ home was identifi ed as 14.2% compared to the cost of building a conventional home without energy-effi cient features. The main barriers that prevent the incorporation of sustainable features into residential developments were identifi ed as cost and lack of developer awareness. Despite this, two-thirds of the respondents felt that consumers would be willing to pay up to a 10% premium for an environmentally friendly home. Unfortunately, this falls below the 14.2% premium identifi ed.

In this study, current and future anthropogenic hydrogen (H 2 ) emissions to the atmosphere from technological processes are assessed. Current emissions are dominated by the direct exhaust gas of road-based motor vehicles and losses during the industrial production of H 2 from fossil-fuels. H 2 emissions from transportation are estimated at 4.5 Tg for 2010. An additional ∼0.5–2 Tg H 2 are estimated to be lost to the atmosphere from industrial processes in 2010. In 2020, emissions from transportation are estimated at approximately 50% of those in 2010. Future emissions will occur as losses along the entire production, distribution, and end-use chain, including emissions from H 2 fuel cell vehicles (FCVs). In 2050, overall anthropogenic H 2 emissions will only approach current levels at high-end loss rates; direct emissions from transportation are expected to be significantly lower than current levels. In 2100, an average loss rate of 0.5% would result in overall H 2 emissions exceeding current levels even with no net H 2 emissions from FCVs. However, based on an average loss rate of 0.1%, H 2 emission factors from FCVs on the order to 120–170 mg km −1 are projected to result in overall anthropogenic H 2 emissions similar to 2010 levels.

Molecular hydrogen (H2), its isotopic signature (deuterium/hydrogen, δD), carbon monoxide (CO), and other compounds were studied in the exhaust of a passenger car engine fuelled with gasoline or methane and run under variable air-fuel ratios and operating modes. H2 and CO concentrations were largely reduced downstream of the three-way catalytic converter (TWC) compared to levels upstream, and showed a strong dependence on the air-fuel ratio (expressed as lambda, λ). The isotopic composition of H2 ranged from δD = −140‰ to δD = −195‰ upstream of the TWC but these values decreased to −270‰ to −370‰ after passing through the TWC. Post-TWC δD values for the fuel-rich range showed a strong dependence on TWC temperature with more negative δD for lower temperatures. These effects are attributed to a rapid temperature-dependent H-D isotope equilibration between H2 and water (H2O). In addition, post TWC δD in H2 showed a strong dependence on the fraction of removed H2, suggesting isotopic enrichment during catalytic removal of H2 with enrichment factors (ε) ranging from −39.8‰ to −15.5‰ depending on the operating mode. Our results imply that there may be considerable variability in real-world δD emissions from vehicle exhaust, which may mainly depend on TWC technology and exhaust temperature regime. This variability is suggestive of a δD from traffic that varies over time, by season, and by geographical location. An earlier-derived integrated pure (end-member) δD from anthropogenic activities of −270‰ (Rahn et al., 2002) can be explained as a mixture of mainly vehicle emissions from cold starts and fully functional TWCs, but enhanced δD values by >50‰ are likely for regions where TWC technology is not fully implemented. Our results also suggest that a full hydrogen isotope analysis on fuel and exhaust gas may greatly aid at understanding process-level reactions in the exhaust gas, in particular in the TWC.

Objective: To examine whether measures of neighbourhood economic deprivation, social disorganisation, and acculturation explain homicide mortality differentials between Mexican Americans, non-Hispanic black Americans, and non-Hispanic white Americans, net of individual factors. Design: Prospective study, National Health Interview Survey (1986–1994) linked to subsequent mortality in the National Death Index (1986–1997). Setting: United States of America. Participants: A nationally representative sample of non-institutionalised Mexican Americans, non-Hispanic black Amricans, and non-Hispanic white Americans, aged 18–50 at the point of interview. Analysis: Cox proportional hazard models estimate the risk of death associated with various neighbourhood and individual factors. Main results: Both individual and neighbourhood risk factors partially account for race/ethnic disparities in homicide. Homicide mortality risks are between 20% and 50% higher for residents of areas that have economic inequality of 0.50 or greater based on the coefficient of variation, or where 4% or more of the residents are Mexican American, 10% or more of the residents are non-Hispanic black, or 20% or more of the households are headed by single parents (p⩽.05). But residents of areas where 10% or more of their neighbours are foreign born have 35% lower mortality risks than people living in areas with fewer foreign born people (p⩽0.05). These differences persist even after controlling for individual level risk factors. Conclusions: The findings support economic deprivation, social disorganisation, and acculturation theories, and suggest that both neighbourhood and individual risk factors affect race/ethnic differences in homicide mortality. Public health policies must focus on both individual and neighbourhood factors to reduce homicide risks in vulnerable populations.

Conjugated polymers, which, when doped, display metallic levels of conductivity, are materials in which there is considerable interest, with possible applications in optoelectronics, rechargeable batteries and electrical screening. These polymers have been studied by two scanning probe techniques. Molecular images of undoped poly( p -phenylene vinylene) (PPV) and poly(3-hexylthienylene) (P3HT), deposited on gold, silver and graphite substrates, have been obtained by scanning tunnelling microscopy, with local alignment of chains observed on the surface. Results from photon emission STM are also presented, and these provide some spectroscopic information about the band gaps and gap states of the polymers, and about plasmon states of the metal substrates.

The purpose of this report is to compare a computed tomography (CT) injury severity scale for hepatic and splenic injury with the following outcome measures: requirement for surgical hemostasis, requirement for blood transfusion and late complications. Sixty-nine children with isolated hepatic injury and 53 with isolated splenic injury were prospectively classified at CT according to extent of parenchymal involvement. Clinical records were reviewed to determine clinical outcome. Ninety-seven children (80%) were managed non-operatively without transfusion. One child with hepatic injury required surgical hemostasis, and 17 (25%) required transfusion of blood. Increasing severity of hepatic injury at CT was associated with progressively greater frequency of transfusion (P = 0.002 by chi 2-test). One child with splenic injury underwent surgery and eight (15%) required transfusion of blood. Splenic injury grade at CT did not correlate with frequency (P = 0.41 by chi 2-test) or amount (P = 0.35 by factorial analysis of variance) of transfusion. There was one late complication in the nonsurgical group. A majority of children with hepatic and splenic injury were managed non-operatively without requiring blood transfusion. The severity of injury by CT scan did not correlate with need for surgery. Increasing grade of hepatic injury at CT was associated with increasing frequency of blood transfusion. CT staging was not discriminatory in predicting transfusion requirement in splenic injury.

Summary Images of the conjugated polymers poly(p-phenylene vinylene) and poly(3-hexylthienylene) deposited on gold, silver and graphite have been obtained using a Nanoscope II STM in air at room temperature. Areas of semi-crystalline structure have been observed, with local alignment of chains on the surface.

The energy intake of a group of 30 patients with cystic fibrosis (CF) was compared with 30 age/gender matched controls. Dietary intakes were determined from 7-day weighed food records using computerized food composition tables. Absolute energy intakes of the CF group were comparable to those of the controls: when expressed as energy per kg body-weight, the CF group consumed approximately 34% more energy (P

Energy intakes and basal metabolic rates were determined in 26 children receiving chemotherapy in remission from acute lymphoblastic leukaemia or solid tumours and 26 healthy controls matched for age and sex. Body weight and height on the two groups were comparable, although one patient was stunted (height for age) and three others wasted (weight for height). Energy intake in the patients at 7705 kJ/day (1842 kcal) and controls at 7773 kJ/day (1866 kcal)) and basal metabolic rate (BMR) in the patients at 4873 kJ/day (1172 kcal) and controls 4987 kJ/day (1196 kcal) for the two groups were not significantly different. Although the energy intake:BMR ratio for both groups was 1.59, the range of values for the patient group was large (0.96-2.73) and appeared to be greater than that observed in the control group (1.23-2.46). These results demonstrated that during this period of chemotherapy there was no evidence of raised energy expenditure at rest or reduced energy intake in the patient group. No indication of undernutrition in the patients as a group was evident, although some individuals might require further clinical nutritional assessment.

The genotype-dependent response of mice to ethanol has been well documented. Cytosolic acetaldehyde dehydrogenase (ALDH-2) increases in some strains while decreasing in others with ethanol treatment. Further work suggests that the mRNA for ALDH-2 (Ahd-2 mRNA) levels are altered following ethanol feeding in a strain-dependent fashion. This report identifies differences in Ahd-2 at the genomic DNA level among different strains of mice. Restriction fragment length polymorphisms (RFLPs) associated with the Ahd-2 locus were found for the restriction enzymes EcoRI, HindIII, Pst I and Rsa I. The mouse strains included in this study could be categorized into two groups based on their overall Ahd-2 associated DNA banding patterns. Strains C57BL/6J, C57BL/6J*, C57BL/10J and BALB/c form group 1 while strains C3H/HeJ, C3H/HeSnJ, 129/ReJ, Csb, SW and DBA/2J form group 2. With the exception of BALB/c, group 1 represents alcohol preferring strains while group 2 are alcohol avoiding strains. Additional work will be required to determine the physiological significance (if any) of these RFLPs and their possible relationship to ethanol preference and avoidance.

41 short normal children were randomly allocated either to daily injections of growth hormone (rhGH) at 30 lU/m 2 per week or to no treatment. 6 months of rhGH therapy produced up to 76% loss of fat mass and up to 25% increase in lean body mass (LBM). These changes were significantly different from those in the untreated group. LBM was the main determinant of resting energy expenditure (REE) expressed as kJ/24 h. REE expressed as kJ/kg LBM per 24 h correlated negatively with height, which was responsible for 66% of the variance in REE kJ/kg LBM per 24 h. Short children therefore expend more energy than tall childen in fulfilling basic metabolic needs. After 6 months REE kJ/24 h increased significantly in treated children. However, treated children did not differ significantly from untreated children in REE kJ/kg LBM per 24 h. rhGH does not therefore seem to have a specific effect upon REE. The possibility that rhGH produces profound metabolic effects should limit its use in otherwise healthy children until the mechanism of action is more clearly elucidated.

Parker's rat coronavirus (PRC) is a naturally-occurring viral infection of the laboratory rat. On the first passage, ATCC strain 8190 of PRC replicated in L-2 cells. Using the tenth passage of PRC in L-2 cells, the characteristics of the virus were compared with previous studies of sialodacryoadenitis virus (SDAV) replicated in L-2 cells. Based on light and immunofluorescence microscopic examination of control and inoculated cell cultures, PRC-associated CPE was frequently confined primarily to individual cells, and there were relatively few syncytial giant cells. Maximum titers were recovered at 36 h post inoculation (pi). Infectious virus was demonstrated at pH values ranging from 6.0 to 9.0 and a pH of 7.5 was determined to produce the highest titers of PRC. The optimum temperature for viral replication was 33 degrees C. Up to 15 passages of PRC in L-929 cells failed to produce detectable virus. However, after adaptation in L-2 cells (20th passage), PRC replicated to high titers in L-929 cells. Previously, in vitro studies of rat coronaviruses have been hampered by the lack of an identified continuous cell line to replicate these viruses in the laboratory. L-2 cells represent a readily-available continuous cell line that can support the replication of relatively high titers of PRC.

Efficient mucociliary clearance is a function of the physical properties of the mucus coupled to appropriately functioning cilia and may be altered by substances affecting ciliary beat frequency (CBF). Therefore the effect of preservatives on CBF was investigated using a photoelectronic technique. Methyl-p-hydroxybenzoate, propyl-p-hydroxybenzoate, chlorbutol and chlorocresol inhibited beat frequency, an effect which was reversible upon rinsing out the first three compounds but not chlorocresol. The effect of chlorhexidine and phenylmercuric borate on CBF was complicated by an interaction with chloride ions in the media used. EDTA did not appear to be ciliotoxic, while the effect of benzalkonium chloride was variable. Thiomersal halted ciliary beating after 40–100 min. Mucociliary clearance may also be affected by an alteration of the physical properties of the mucus layer, therefore the effect of each compound on the rheological properties of purified pig gastric mucus glycoprotein was investigated. None of the preservatives significantly altered the viscoelastic properties of the gel, measured using dynamic techniques.

Endothelin-1 (ET-1) is present in murine and human skin and causes itch (pruritus) when injected in humans. This behavioural study examined the scratch reflex evoked by ET-1 in mice.An automated detector was used to determine whether ET-1 causes reflex scratching, the behavioural correlate of itching, in BALB/c mice. Selective agonists and antagonists were used to probe the ET receptor(s) involved.ET-1 evoked dose-related reflex scratching lasting up to 20 min following intradermal injection (0.1-100 ng; 0.04-40 pmol). The ED(50) for ET-1 induced scratching was 2.1 ng and desensitization occurred with cumulative dosing. High doses of the ET(B) receptor agonist IRL1620 (10 microg; 5.5 nmol), also caused scratching (ED(50) 1.3 microg, 0.7 nmol). The ET(A) receptor antagonist BQ123 significantly reduced scratching evoked by ET-1 and IRL 1620, suggesting that both agonists caused scratching via an ET(A) receptor-dependent mechanism. The ET(B) receptor antagonist BQ788 significantly reduced scratching evoked by IRL1620 but had no effect on scratching evoked by ET-1. This indicated that activation of ET(B) receptors by high doses of ET(B) agonist, but not ET-1, can trigger scratching.ET-1 is a potent endogenous activator of reflex scratching (itch). Mechanisms for ET-induced scratching are considered, including direct action of ET-1 on pruriceptive nerve endings and indirect actions via release of endogenous mediators such as histamine from mast cells. ET-1 and ET(A) receptors, possibly also ET(B) receptors, are potential targets for developing specific anti-pruritic drugs to treat pruritic skin disorders such as atopic dermatitis.

If one drug, B, antagonizes another, A, by producing the opposite physiological effect, the antagonist concentration-effect curves should be affected by the recording system, which limits the range of agonist responses. With pieces of isolated guinea-pig ileum taken from adjacent parts of the same animal, one recorded isotonically, the other isometrically with the same load, the isotonic IC(50) values for (-)isoprenaline opposing carbachol or histamine were lower than the isometric values (P0.01) but there was a significant correlation between them (P0.01): the isotonic curves were steeper (P0.01) and there were wider shifts in IC(50) before increasing the agonist reduced the maximum relaxation. In similar experiments with pieces of rat uterus in oestrus from the same animal, the concentration-effect curves for carbachol opposed by increasing concentrations of (-)isoprenaline or (-)adrenaline had slightly lower EC(50) values with isometric recording but there was a significant correlation (P0.01) with isotonic values. The antagonist effect (ratio of the EC(50) relative to that for the control) was higher with isotonic recording (P0.01 for (-)isoprenaline, P0.025 for (-)adrenaline) and all (27) curves were steeper than the corresponding isometric curve (P0.001). The influence of the method of recording on the results is expected from the narrower operational window and smaller upper limit to relaxation with isotonic recording. A way of obtaining measurements of IC(50) against a standard agonist effect is suggested in an Appendix.

Poor gap junctional intercellular communication (GJIC) has been associated with uncontrolled cell growth and neoplasia. We have successfully propagated normal first trimester invasive extravillous trophoblast (EVT) cells, and have produced premalignant EVT lines after SV40 Tag transformation: RSVT-2 is an uncloned line that is long-lived; RSVT2/C is a clonal line that is immortal. Both are hyperproliferative, hyperinvasive and variably refractory to the anti-proliferative and anti-invasive effects of transforming growth factor beta (TGFbeta). Possible changes in gap junctions during the transition of normal invasive EVT cells to the premalignant stage were examined by comparing expression of connexin proteins (by immunolabeling for Cx26, Cx32, Cx40, Cx43), and mRNA (by Northern blot with cDNA probes for Cx26, Cx32, Cx43), and functional GJIC (by dye transfer using the preloading method) in normal parental EVT cells and their SV40 Tag transformants. Results from immunofluorescence and Northern blot analysis revealed that, of the panel of connexins examined, only Cx43 was variably expressed in these cell lines in vitro. Expression of Cx43 protein and mRNA was abundant in normal EVT cell line HTR8, reduced in long-lived RSVT-2 cells and undetectable in immortalized RSVT2/C cells. GJIC, as measured by dye transfer between donor and recipient cells, was also similarly reduced in recipient RSVT-2 cells, and drastically reduced in RSVT2/C cells, irrespective of whether the dye donor was of the same cell type (homocellular coupling) or HTR8 cells (heterocellular coupling). Treatment with TGFbeta reduced Cx43 mRNA expression as well as GJIC in normal EVT cells, but not in the SV40 Tag transformants. Our findings suggest that downregulation of connexins with the resultant impairment in GJIC is an early event in tumor progression, as observed in the premalignant SV40 Tag transformants.

Invasion of the uterus by first trimester human placental extravillous trophoblast (EVT) cells depends on mechanisms shared by malignant cells. However, unlike tumor invasion, trophoblast invasion of the uterus is stringently controlled in situ by local molecules such as transforming growth factor (TGF)beta. Since EVT cells possess active invasion-associated genes but are nontumorigenic, our objective was to induce premalignant and then malignant phenotype into a normal EVT cell line in order to identify the molecular basis of tumor progression. Simian virus 40 large T antigen (SV40 Tag) was introduced into a normal human first trimester invasive EVT cell line, HTR8, established in our laboratory. Since the HTR8 line has a limited in vitro lifespan of 12-15 passages, SV40 Tag-transformed cells were selected on the basis of extended lifespan. A long-lived line, RSVT-2, was produced and an immortalized subclone, RSVT2/C, was further derived under a forced crisis regimen. We examined transformation-induced alterations in proliferative and invasive abilities, responses to the invasion and proliferation-regulating growth factor TGFbeta and changes in gene expression for invasion-associated enzymes or enzyme inhibitors. RSVT-2 and RSVT2/C cell lines were hyperproliferative and hyperinvasive when compared with the parental HTR8 cell line. They were also variably resistant to the anti-proliferative and anti-invasive signals from TGFbeta. Since both cell lines remained non-tumorigenic in nude mice, these properties indicate that they attained a premalignant phenotype. Both cell lines showed reduced expression of tissue inhibitor of metalloproteases (TIMP)-1, while TIMP-2 and plasminogen activator inhibitor (PAI)-I expression was was also reduced in RSVT2/C cells, thus contributing to their hyperinvasiveness. Their resistance to the anti-invasive action of TGFbeta was explained by the failure of TGFbeta to upregulate TIMPs and PAI-I, in contrast to the TGFbeta-induced upregulation noted in parental HTR8 cells.

1. Experiments carried out on guinea-pig isolated ileum with carbachol as agonist and diphenyl-acetoxyethyl-dimethyl-ethyl-ammonium (DADMEA) bromide as antagonist gave results which fit the theoretical relation between fractional inhibition (Q) of the effects of an agonist ([A]) and the concentration of a competitive antagonist ([B]): this also involves the Hill coefficient (logistic slope factor, P) for the agonist concentration-response curve and the degree of agonist stimulation, [A]/[A]50, where [A]50 produces a half-maximum response. 2. Values of IC50 and an exponent, P', can be obtained by fitting Q to [B] using a logistic approximation to the relation. Both P' and IC50 should be greater with higher agonist stimulation but the increase in P' may be masked by errors in extreme values of Q. Estimates of IC50, however, invariably increased with higher agonist stimulation but with a steep concentration-response curve (P1) and low agonist stimulation ([A]/[A]501, IC50 can be less than KD. 3. KD was calculated from the results in three ways; (i) by a least-squares fit of Q to [B] using the values of P and [A]/[A]50 calculated from the control concentration-response curve; (ii) from the value of IC50 for each line and the values of P and [A]/[A]50 and (iii) by using the agonist concentration-response curve to calculate the dose-ratio and estimate of KD for each in the presence of the antagonist. The methods gave similar results (nM: 11 experiments), 12.4 +/- 1.1 (i), 11.7 +/- 0.9 (ii), 14.8 +/- 1.6 (iii) but there are advantages in using methods (i) or (ii) rather than (iii). 4. The method by which KD is calculated is less important than the experimental design: the plan used in this work, with alternative small and large responses from the tissue, is very suitable for estimating KD with low concentrations of antagonists and small dose-ratios. Although it is not a sensitive test for competitive behaviour because only a small range of concentrations of antagonist is tested, the estimate of affinity should be free from complications involved in the use of higher concentrations of antagonist (and agonist) and the nature of the antagonism can always be checked by doing further experiments in the presence of a known competitive antagonist.

The objective of this study was to determine the efficacy of two-dimensional and contrast echocardiograms to identify venous cannula position. Sequential sampling of 20 infants was evaluated by contrast echocardiography after meeting institutional criteria for extracorporeal life support. Each infant was placed on venovenous extracorporeal life support using a double-lumen cannula. After surgical placement was thought to be satisfactory, optimal two-dimensional images of the cannula were obtained via a subxiphoid or apical view and 2 ml agitated normal saline were injected rapidly into the nearest infusion port. Patient demographics and mixed venous saturations were noted. Distance of the venovenous cannula to tricuspid valve and distance of the venovenous cannula from the intra-atrial septum was recorded. Echocardiograms were available for review on 18 of the 20 patients. Position of the venovenous cannula in relationship to the tricuspid valve was as follows:5 mm (8); 5-10 mm (5);10 mm (5). Mixed venous saturations decreased, which indicated less recirculation when the orientation of the tip of the cannula was toward the lateral wall in those who required repositioning. The authors conclude that two-dimensional and contrast echocardiography aid in the positioning of the venovenous cannula. Satisfactory position is approximately 5 mm from the tricuspid valve, with orientation toward the lateral wall of the right atrium.

The aim of this study was to investigate the angiogenic response to exogenously administered basic fibroblast growth factor (FGF-2) in normal and ischemic skin, using the hairless mouse ear microcirculatory model. The hairless mouse ear is a well-established model for in vivo studies of skin microcirculation. Using this model, angiogenesis- and angiogenesis-associated changes in the microcirculation can be directly and continuously viewed and quantified in a variety of different experimental settings. To create ischemia in the mouse ear, all but one of the three to four feeding vessels nourishing the ear were ligated 3 days prior to a local subdermal injection of FGF-2 (9.3 + 1-0.5 mm/mm2) or saline into the dorsum of the ears. Angiogenesis was quantified by direct observation, at high magnification, of the injection site where increases in total vessel length (TVL) were measured repeatedly over 18 days following injection. We found a significant (P0.01) increase in TVL in normal and ischemic ears injected with FGF-2. Saline injection also induced a significant increase in TVL in ischemic ears. However, the angiogenic response to FGF-2 in ischemic ears was significantly stronger than saline alone in ischemic ears or saline or FGF-2 in normal ears. This response could be used clinically to accelerate angiogenesis and thus increase perfusion in ischemic tissue.

The effects of three structural types of nipecotamides and their stereoisomers on collagen-induced aggregation and intraplatelet ionized calcium ([Ca2+]i) rise in human platelets were evaluated using aequorin as the [Ca2+]i indicator. The orders of potencies of racemic nipecotamides were different when collagen was the agonist compared with those obtained using ADP. It is suggested that in addition to their earlier hypothesized interactions with platelet anionic phospholipids of the plasma and organelle membranes, nipecotamides may, in addition, act at other receptor sites. In general, the inhibition of collagen-induced aggregation paralleled their inhibitory effects on the rise of [Ca2+]i. The compounds were stereoselective in inhibiting aggregation as well as [Ca2+]i rise. The meso diastereomers of I and II were more potent than the corresponding enantiomeric pairs. A single [Ca2+]i peak was noticed when the incubate contained 1.0 mM extracellular calcium [Ca2+]o. On the other hand a biphasic [Ca2+]i rise was noticed when the nominally Ca(2+)-free buffer contained 75 microM ethylene glycol tetraacetate (EGTA). The first peak corresponded with platelet shape change, suggesting Ca2+ discharge from internal stores, and the second, with aggregation. The second peak may reflect either Ca2+ flux across the plasma membrane or aequorin leak from internal cellular locations or from the canicular system. Inhibition of the rise in intraplatelet Ca2+ appears to be associated with the platelet aggregation-inhibitory actions of nipecotamides.

Genitourinary rhabdomyosarcoma in females usually originates in the vagina or uterus, but rarely the vulva. The authors present a case of rhabdomyosarcoma originating in the clitoris. A 4-year-old with an alveolar rhabdomyosarcoma of the clitoris was treated with radical clitorectomy, radiation, and chemotherapy. Follow-up at 3 years showed no active disease.

C6 glioma cells do not express the gap junction protein connexin32 or its corresponding mRNA. Very low levels of connexin43 protein and mRNA, as well as weak intercellular coupling, have been detected. Studies investigating the role of gap junctions in cell proliferation and tumorigenesis have shown that C6 cells transfected with connexin43 have increased levels of intercellular coupling and reduced cell growth (D. Zhu et al., Proc. Natl. Acad. Sci. USA, 88:1883-1887, 1991). To determine whether this growth inhibition is observed with other connexins, a full-length cDNA for connexin32 was used to transfect C6 cells. A number of transfected clones, expressing various levels of connexin32 mRNA, were obtained. Further analysis of several of these clones has shown that they have a corresponding increase in both the amount of connexin32 immunoreactivity and intercellular coupling. Thus, transfection of the C6 glioma cell line with connexin32, a gene which is normally expressed in the rat brain but not in C6 cells, produces both a functional mRNA and protein. Growth of the transfected clones was reduced in vivo. In vitro, growth of the various clones was not correlated to either levels of connexin32 expression or intercellular coupling. This is in contrast to findings in the previous study, in which cell growth was reduced in response to connexin43 expression both in vivo and in vitro in the transfected cells. These clones provide a unique system to study the role of gap junctions in cell proliferation and other tumor characteristics.

Measurements of H 2 at the high-altitude site of Jungfraujoch, Switzerland are reported upon for the period of August, 2005–November, 2009. The time series consists of measurements that are primarily representative of free tropospheric background conditions. Highest background H 2 mixing ratios were observed in May, while the lowest were observed in November. The mean seasonal H 2 peak-to-trough amplitude of 21 parts per billion (ppb, 10 -9 dry air mixing ratio) at Jungfraujoch was considerably less than at other stations at similar latitudes and the seasonal minimum in November was comparatively delayed. These differences are primarily attributed to a dampening and delay of the surface soil sink signal during its vertical propagation to the free troposphere. Excess (mixing ratio minus corresponding baseline value) H 2 (ΔH 2 ) and excess CO (ΔCO) displayed no significant correlation. This lacking correlation is attributed to H 2 removal by soil during transport to Jungfraujoch, thereby significantly altering the ΔH 2 /ΔCO ratio from traffic combustion sources, which is the largest source of anthropogenic H 2 influencing measurements at Jungfraujoch. DOI: 10.1111/j.1600-0889.2010.00509.x

A reflection grating wavelength demultiplexer fabricated on planar silica-on-silicon is investigated for use both as a multi-channel demultiplexer and as part of a fast single- wavelength selector device. Design and characterization of a preliminary grating device which resolves 128 channels with 0.85 nm channel spacing are presented in this paper.

In an effort to develop compounds with high antithrombotic activity and minimal toxicity, our laboratory has synthesized a number of nipecotamides. The effectiveness of one of these compounds, alpha,alpha'-bis[3-(N,N-diethylcarbamoyl)piperidino]-p-xylene dihydrobromide (A-1), in inhibiting both in vitro and in vivo platelet aggregation is reported here, along with its acute toxicity. The IC50 of A-1 in in vitro ADP- and PAF-induced platelet aggregation was 44.5 microM and 21.2 microM, respectively. Suppression of intraplatelet [Ca2+] is suggested as a likely mediator of the aggregation-inhibitory properties of A-1, since both the release of cytosolic Ca2+ and the influx of extracellular Ca2+ were decreased. The ED50 of A-1 in protecting mice against thromboembolism induced by a collagen-epinephrine challenge was 164 mumol/kg. The measurement of the acute toxicity of this compound as the LD50 was 691 mumol/kg, with the therapeutic index being 4.2. These data indicate that compounds in this family hold promise as clinically effective antithrombotic agents.

In order to determine whether or not there was a relationship between disorders of growth in children suffering from asthma and either increased resting energy expenditure or inadequate energy intake, a group of 34 children suffering from perennial symptoms were studied. A control group matched with the asthmatic children for sex and fat free mass were similarly studied. The children kept seven day records of weighed food intake. Basal metabolic rate was measured on one occasion in the fasted state by means of indirect calorimetry using the ventilated hood technique. The asthmatic children kept a 28 day record of peak expiratory flow rates, asthma symptoms, and medication usage. The asthmatic children expended significantly more energy at rest than their matched controls in absolute terms (14%). There was no correlation between height or height SD score and any parameter of energy balance. The causes of these finding are as yet speculative.

One hundred eighty pediatric patients with suspected appendicitis were prospectively examined with graded compression ultrasonography (US) to assess the sensitivity, specificity, and accuracy of graded compression US in the diagnosis of appendicitis in children and to compare those results with results of clinical assessment in the diagnosis of this disorder. Patients were assigned to one of three groups prior to US based on the clinical level of confidence that appendicitis was present and on the planned management decision. Of 141 patients in the low- and intermediate-clinical risk categories, 20 (14%) had appendicitis: US had a sensitivity of 100%, specificity of 97%, and accuracy of 97% in these two groups. Of 39 patients in the high-clinical risk category, 32 (82%) had appendicitis: US had a sensitivity of 81%, specificity of 86%, and accuracy of 82%. Of 52 patients with surgically proved appendicitis, the initial management decision was to discharge to home or admit for observation and further testing in 18 (35%). Results at US were positive for appendicitis in all 18 patients in the latter two categories.

The advent of improved obstetrical care and innovative advances in fetal diagnostic techniques have mandated multidisciplinary approaches to pregnancies that are complicated by prenatally diagnosed defects. The concept of fetal therapy, previously limited to in utero transfusions for Rh disease and induction of lung maturation, has now been extended to open fetal correction of congenital hydronephrosis, diaphragmatic hernia, and sacrococcygeal teratoma. Additional inquiries are now being made into the possibility of hematopoietic stem cell transplantation in utero as well as prenatal gene therapy. Continued investigation into these therapeutic interventions largely revolves around improving their efficacy and guaranteeing the safety of the mother and her unborn child.

Four hybridoma cell lines were grown and supported, using fully integrated bioreactor systems. Each system, consisting of a sterile, disposable, hollow fiber bioreactor, was maintained by totally automated control instrumentation. Characterization of cell line viability, productivity, and scalability was established by employing hollow fiber cartridges of three different sizes. The efficacy of the system was evaluated in terms of operational requirements, process control, antibody production, and end product purification as compared to alternative cell culture techniques.

During a 48-month period, 2,248 children (aged less than 15 years) were consecutively admitted to a regional pediatric trauma center with blunt trauma (ICD-9-CM code greater than or equal to 800). Fifty-four children (2.4%) had injury to the pelvic circle, as diagnosed by radiographic examination; 13 of these children had concomitant abdominal or genitourinary (GU) injury. Contingency table analysis and stepwise logistic regression were used to determine the best predictors of abdominal injury. The mean age of the children was 8.6 years. Eighty-nine percent of the injuries were motor-vehicle related (59% pedestrian; 30% crash occupant). Nine children (17%) required transfusions of packed red blood cells; 9 children (17%) required surgery. There were 6 deaths in this group, a mortality rate of 11.1%. The most common fracture sites in the pelvis were the pubic rami (59%), ilium or pelvic rim (17%), and the sacrum (6%). Ten children (19%) had multiple pelvic fractures. Location of fracture was strongly associated with the probability of abdominal injury: 80% of children with multiple pelvic fractures had concomitant abdominal or GU injury, compared with 33% with fracture of the ilium or pelvic rim, and 6% with isolated pubic fractures (p less than 0.001). The variables that best predicted abdominal or GU injury using a backward-elimination, stepwise logistic model were the presence of multiple pelvic fractures (p less than 0.002) and unweighted Revised Trauma Score (p less than 0.05); age of child, systolic blood pressure, respiration rate, Glasgow Coma Scale score, and mechanism of injury were not predictive.(ABSTRACT TRUNCATED AT 250 WORDS)

Stool energy losses and the sources of energy within the stool were determined in 20 healthy controls and 20 patients with cystic fibrosis while on their habitual pancreatic enzyme replacement treatment. Stool energy losses were equivalent to 3.5% of gross energy intake in healthy children (range 1.3-5.8%). Despite a comparable gross energy intake, stool energy losses were three times greater in patients with cystic fibrosis than controls averaging 10.6% of gross energy intake (range 4.9-19.7%). Stool lipid could account for only 29% and 41% of the energy within the stool in controls and patients with cystic fibrosis respectively and was poorly related to stool energy. Approximately 30% of the energy within the stool could be attributable to colonic bacteria in both the healthy children and patients with cystic fibrosis. These results suggest that stool energy losses in healthy children are relatively modest but that even when patients with cystic fibrosis are symptomatically well controlled on pancreatic enzyme replacement, raised stool energy losses may continue to contribute towards an energy deficit sufficient to limit growth in cystic fibrosis. As the energy content per gram wet weight remains relatively constant (8 kJ/g), stool energy losses may be estimated from simple measurements of stool wet weight.

Mice (Mus musculus) from three genetic strains with variable responses to ethanol challenge (BALB/c, C57BL/6J and 129/ReJ) were used to evaluate the effect of ethanol feeding on hepatic mRNA specific to the two primary enzymes of ethanol metabolism; alcohol dehydrogenase (ADH; E.C. 1.1.1.1) and aldehyde dehydrogenase (ALDH; E.C. 1.2.1.3). Adh-1 (ADH) and Ahd-2 (ALDH) specific mRNA were evaluated on the livers of ethanol-fed mice and from their age, sex and genotype matched controls (using an isocaloric liquid diet). C57BL/6J (alcohol resistant) mice show a significant (approx. 200%) increase in ADH-1 mRNA levels after ethanol treatment, compared to their matched controls. BALB/c (alcohol sensitive) mice have approximately a 20% increase with ethanol treatment while 129/ReJ (alcohol sensitive) mice show a slight reduction in the ADH-1 specific mRNA following ethanol feeding. A strain-specific pattern is also apparent in the AHD-2 mRNA as a result of ethanol feeding in the experimental animals. C57BL/6J mice have an increase and BALB/c mice show no apparent change in the AHD-2 mRNA. 129/ReJ mice fed an ethanol diet, on the other hand, appear to have a decrease in the level of AHD-2 hepatic mRNA as compared to their matched controls. The relative mRNA levels of the two genes correlate well with the respective enzyme activity levels, but for mice on the control diet only. Ethanol feeding, which causes an apparent reduction in hepatic ADH enzyme activity in BALB/c and 129/ReJ and an apparent increase in ALDH activity in C57BL/6J (under the experimental protocols used) also alters the mRNA levels specific to the two genes. However, changes in the mRNA levels after ethanol feeding cannot be directly related to the changes seen in enzyme activity. The observed steady state level of AHD-2 mRNA and the increase in ALDH activity after ethanol feeding, which is unique to C57BL/6J mice, is expected to offer a faster clearance (metabolism) of acetaldehyde, the toxic metabolite, and may be responsible for, or contribute to, the relative resistance of this strain to ethanol.

Wistar rats [Cr1:(WI)BR] were inoculated intranasally with approximately 10(3) median mouse lethal infective doses of sialodacryoadenitis (SDA) virus. Animals were subsequently selected at random, removed to a separate isolation room, and reinfected with SDA virus at 3, 6, 9, 12 or 15 months. Pre- and postinoculation serum samples were collected from all animals during the course of the study and evaluated for antibody titers to SDA virus. All experimental, control and sentinel animals, following inoculation with SDA virus, were necropsied and examined for lesions consistent with SDA. Salivary gland lesions were minimal to absent in rats reinfected with SDA virus for up to 12 to 15 months after the initial exposure and minimal to moderate in the respiratory tract at 12 or 15 months. SDA-associated lesions were extensive in age matched control animals examined at each time period of reinfection with SDA virus. Thus, prior exposure to SDA virus did protect against the development of typical salivary gland lesions for up to 15 months. Recovered animals were evaluated for their ability to transmit the virus following reinfection. Rats reinfected at 6 or 9 months were infectious to their naive cage mates. The results indicate that reinfection with homologous rat coronavirus can occur as early as 6 months after the initial infection, and such rats can transmit the infection to contact controls.