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Studies on plating efficiency and estimation of viability of suspensions of Paracoccidioides brasiliensis yeast cells.

Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimated viability by means of vital dyes. Methylene blue Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans. Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium. In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25%. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens.

Acridine Orange↗

Replating efficiency of metastatic melanoma cells from lymph node and subcutaneous sites does not predict patient survival.

The efficiency of replating of cells from primary colonies grown in semisolid medium has been used to detect and quantitate self-renewal in vitro. A positive correlation has been found by others between the replating efficiency of cells from myelogenous leukemia and patient survival. In the current study we measured primary and secondary replating efficiency of metastatic melanoma cells from subcutaneous tissues or lymph nodes of twelve patients and related these results to patient survival from time of biopsy. No relationship was found between primary and secondary plating efficiency nor for primary or secondary replating efficiency and survival. These results suggest that colony-forming melanoma cells grown under anchorage-independent conditions do not identify a stem cell population important for survival distinct from highly proliferative cells. These studies do not, however, rule out the possibility that a non-clonogenic transitional cell population exists in the tumor.

Cell Division↗

Effects of feeder layers made of human, mouse, hamster, and rat cells on the cloning efficiency of transformed human cells.

The effect of feeder layers on cloning efficiency of transformed human cells was investigated. Embryonic human skin or lung fibroblasts; adult human skin fibroblasts; early passage cells from embryos of mouse, rat, and hamster; established mouse cell lines; 3T3 and 10T1/2 were used as feeder layers after they were lethally exposed to Co-60 gamma-rays at 3,000 rad. As test cells to study the effect of feeder layers on cloning efficiency, WI-38 CT 1 cells transformed in vitro by Co-60 gamma-rays and HGC cells cultured from a human gastric cancer were used. The effect of feeder layers on the cloning efficiency of the test cells was dependent on cell density of feeder layer cells, sources of the feeder layer cells, and kinds of test cells. An optimal density of feeder cells produced cloning efficiencies 3 to 15 times higher than in cultures without a feeder layer. Generally, high density of cells in feeder layers decreased the cloning efficiency of the test cells, presumably owing to contact inhibition of growth and depletion of essential nutrients by the feeder layer cells. Regarding the effect of the feeder layers made of human fibroblasts, there were no significant differences in population doubling levels; tissue origins of fibroblasts, or fibroblasts derived from normal individuals, patients with cancer, or with a genetically high familial incidence of cancer, hereditary adenomatosis of the colon and rectum.

Animals↗

Effects of space flight, clinorotation, and centrifugation on the substrate utilization efficiency of E. coli.

Cultures of Escherichia coli grown in space reached a 25% higher average final cell population than those in comparably matched ground controls (p<0.05). However, both groups consumed the same quantity of glucose, which suggests that space flight not only stimulated bacterial growth as has been previously reported, but also resulted in a 25% more efficient utilization of the available nutrients. Supporting experiments performed in "simulated weightlessness" under clinorotation produced similar trends of increased growth and efficiency, but to a lesser extent in absolute values. These experiments resulted in increases of 12% and 9% in average final cell population (p<0.05), while the efficiency of substrate utilization improved by 6% and 9% relative to static controls (p=0.12 and p<0.05, respectively). In contrast, hypergravity, produced by centrifugation, predictably resulted in the opposite effect--a decrease of 33% to 40% in final cell numbers with corresponding 29% to 40% lower net growth efficiencies (p<0.01). Collectively, these findings support the hypothesis that the increased bacterial growth observed in weightlessness is a result of reduced extracellular mass transport that occurs in the absence of sedimentation and buoyancy-driven convection, which consequently also improves substrate utilization efficiency in suspended cultures.

Biological Transport↗

Genetic variation in the efficiency of nitrogen utilization and photosynthetic activity of flag leaves among the old and modern germplasm of winter wheat.

Genotypic variation in major components of the efficiency of nitrogen utilization and photosynthetic activity of flag leaves among old (released 1881-1963) and modern (released 1969-2003) cultivars of winter wheat was studied in field conditions under varied N fertilization levels (110, 90 and 80 kg N ha-1). Significant genotypic differences were observed for all characters. Their heritabilities ranged from 0.37 to 0.93 and were the lowest for the leaf efficiency of gas exchange, photosynthetic rate, straw N content and the economic index of N utilization efficiency (NUE). Some modern cultivars exhibited an enhanced tolerance to N shortage and several attributes of efficient N utilization (e.g. later senescing and more photosynthetically active flag leaves, increased ability to redistribute N into grains). The genotypes may serve as donors of appropriate characteristics for breeding. The observed cultivar-by-fertilization interactions suggest, however, that evaluations under diverse fertilization regimes may be necessary when searching for improved wheat efficiency and adaptation to less favourable environments.

Adaptation, Physiological↗

Surfactant nebulisation: safety, efficiency and influence on surface lowering properties and biochemical composition.

OBJECTIVE: The objectives of this study were, to select a nebuliser first, that operates safely in a neonatal ventilator setting and, second, that is most efficient. Thirdly, we studied the particle sizes of the surfactant aerosol. Fourthly, we studied where the nebulised surfactant is deposited in the tubing system of the ventilator. Finally, we studied whether nebulisation influences the composition and biophysical properties of surfactant. MEASUREMENTS AND RESULTS: Safety was assessed by measuring "mean airway pressures" in a test lung before, during and after surfactant nebulisation, for three jet nebulisers. The MiniNEB did not alter these pressures, and is thus safe, whereas the other two nebulisers (Intersurgical and Flo-Thru) increased these pressures. The efficiency of nebulisation was assessed by measuring the amount of phospholipid deposited in the test lung. The MiniNEB showed the highest efficiency: 10% versus 1-3% of the other two nebulisers. The particle sizes of surfactant aerosol were assessed by the laser diffraction method. Seventy percent of the particles were 1-5 microns. The deposition of surfactant aerosol in the tubing system was assessed by nebulising surfactant that was labelled with 99mTc Nanocoll. Afterwards the tubing system was imaged using a gamma camera. The majority of surfactant was deposited in the expiratory hose (28%), nebuliser (20%), Y-piece (16%) and expiratory filter (12%). Finally the phospholipid composition, spreading velocity, static and dynamic surface tensions were assessed for the nebulised surfactant and compared to the stock surfactant. In addition, nebulised surfactant was instilled in premature rabbits and tidal volumes were measured to assess the dose-response relation. We found that neither the composition nor biophysical properties had been altered by nebulisation. CONCLUSIONS: The MiniNEB nebulised surfactant safely in a neonatal ventilator setting with respect to airway pressures. The efficiency of nebulisation is low: the majority of the surfactant aerosol is deposited in the expiratory tubing. The surfactant composition and function is not altered by nebulisation. Therefore the nebulisation of surfactant is feasible, but efforts should be made to improve the efficiency of this procedure.

Aerosols↗

The efficiency of chemical vapour generation of transition and noble metals.

The efficiency data from individual reports on chemical vapour generation (CVG) of transition and noble metals are overviewed with respect to the ways that they were obtained. The values derived from direct measurement of analyte in the gaseous phase or obtained from comparison with other sample introduction techniques reflect well the true chemical vapour generation efficiency. The efficiencies reported this way do not exceed 30%. On the other hand, the percentage efficiency values derived from determination of analyte remaining in the waste liquid, usually in the high tens, can substantially overestimate the efficiency by neglecting the analyte retained within the system.

Journal Article↗

Analyte transport efficiencies in electrothermal vaporization for inductively coupled plasma mass spectrometry.

A modified graphite furnace for solid-sampling atomic absorption spectrometry as an electrothermal vaporizer (ETV) was coupled to a Perkin-Elmer/Sciex ELAN 6000 ICP mass spectrometer. The integrals obtained from electrothermal vaporization of aliquots containing As, Cd, Cu, Co, Fe, Mn, Pb, Se, and Zn were compared with those obtained from pneumatic nebulization of the same aqueous standard solution. The pneumatic nebulizer was calibrated by weighing the mass of aqueous aerosol trapped on a filter. With "wet plasma" conditions maintained also for measurements with the ETV and reference signals for analyte signals obtained with the calibrated pneumatic nebulization, the transport efficiency of the ETV system, e.g. the ratio of the analyte amount introduced into the plasma to that amount dosed into the vaporizer, was determined. The transport efficiency of two different tube and interface designs has been evaluated. Investigations with and without the use of trifluoromethane as reactive gas, with different furnace heating rates, and with varying gas flows were performed. In general, the tube equipped with a nozzle led to generally higher transport efficiency than the standard tube. Without trifluoromethane transport efficiencies ranged from 10% to 35% with the standard tube and from 15% to 50% with the nozzle-type tube. With addition of 2 mL min(-1) trifluoromethane to the argon flow of 400 mL min(-1) through the tube, transport efficiencies from 20% to 70% and from 70% to 100% were achieved with the standard and nozzle-type tubes, respectively.

Journal Article↗

Noninvasive determination of myocardial blood flow, oxygen consumption and efficiency in normal humans by carbon-11 acetate positron emission tomography imaging.

The aims of this study were: (1) to measure noninvasively and near simultaneously myocardial blood flow, oxygen consumption, and contractile function and (2) to analyze myocardial energy expenditure and efficiency at rest and during dobutamine stress in normal humans. Dynamic and gated carbon-11 acetate positron emission tomography (PET) imaging was performed in 11 normal subjects. The initial uptake of (11)C-acetate was measured to estimate myocardial blood flow. Oxygen consumption was derived from the monoexponential slope of the (11)C-clearance curve recorded during myocardial washout. ECG-gated systolic and diastolic images were acquired during the peak myocardial (11)C activity to measure left ventricular radius, myocardial wall thickness, and long axis length. Myocardial oxygen consumption and parameters of cardiac geometry were used to determine myocardial energetics and cardiac efficiency by tension-area area analysis. Myocardial blood flow averaged 0. 8+/-0.06 ml min(-1) g(-1) at rest and 1.48+/-0.15 ml min(-1) g(-1) during dobutamine stress. Oxygen delivery and consumption were 151+/-13 and 88+/-15 microl O(2) min(-1) g(-1) at rest and increased to 291+/-31 and 216+/-31 microl O(2) min(-1) g(-1), respectively, during pharmacological stress (P<0.001). Oxygen extraction increased from 59%+/-8% at rest to 76%+/-9% during stress (P<0.001). Mechanical efficiency was 29%+/-6% at rest and 32%+/-6% during dobutamine stress (P=NS) while external work efficiency was 16%+/-6% at rest and increased to 21%+/-4% (P<0.01) during dobutamine stress. Stepwise linear regression analysis identified rate-pressure product and external cardiac work as major correlates of oxygen consumption. In summary, rapid dynamic and gated PET (11)C acetate imaging provides the unique capability to study noninvasively determinants of myocardial energy delivery, expenditure, and efficiency.

Acetates↗

Comparison of three 3' non-coding regions in Schizosaccharomyces pombe expression vectors: efficiencies of transcription termination and mRNA 3'-end formation.

Analysis of an established Schizosaccharomyces pombe episomal shuttle vector suggested that inefficient transcription termination was deleterious to plasmid function. We undertook a study to determine if transcription in the presence and absence of 3'-processing within a vector could affect the ability of the plasmid to transform, transcribe and translate the RNA produced. This report provides an analysis of the effects that three S. pombe 3' non-coding regions have on the transformation and expression efficiencies of a fission yeast plasmid vector. The 3' regions from adh1, act1 and ura4 were tested for their ability to terminate and process adh1 promoter-driven transcription of a lacZ reporter gene. Differences between the 3'-processing sequences were observed, with transcription termination mediated by the ura4 3' region being more efficient than termination by the 3' regions of adh1 and act1. We show that plasmids containing inefficient transcription termination signals result in readthrough transcription and reduced transformation efficiencies. In addition, the readthrough transcripts containing 3' non-coding regions show impaired translation efficiencies. We describe an S. pombe vector (pURAS) with a high transformation efficiency that directs the production of highly translatable, discrete-sized transcripts.

Gene Dosage↗

Establishment of a highly efficient transformation system for pepper (Capsicum annuum L.).

Application of modern genetic manipulation has been limited in pepper ( Capsicum annuum L.) due to the lack of an efficient transformation system. Following the development of an efficient protocol for in vitro regeneration of pepper cotyledons, we investigated the key factors affecting transformation and established a highly efficient genetic transformation system using the pepper cotyledon as starting material. In this system, cotyledon explants are preconditioned for 2 days on kanamycin (km)-free DM1 medium [Murashige and Skoog (MS) salts/Gamborg B5 vitamins basal medium supplemented with 20 g/l sucrose, 5,000 mg/l DJ nutrients and a hormone combination of 1.0 mg/l indoleacetic acid (IAA) and 5.0 mg/l 6-benzyladenine (BA) solidified with 0.7% agar, pH 5.8], followed by co-cultivation with Agrobacterium tumefaciens on DM1 for 2 days and delay selection on DM1 with 500 mg/l carbenicillin (carb) for 2 days. The explants are then placed on DM1 containing 10 mg/l AgNO(3), 50 mg/l km-sulfate and 500 mg/l carb. After 4-5 weeks, the explants with buds are transferred to EM1 medium (MS salts/Gamborg B5 vitamins basal medium supplemented with 20 g/l sucrose, 5,000 mg/l DJ nutrients, 10 mg/l AgNO(3) and a hormone combination of 1.0 mg/l IAA, 3.0 mg/l BA and 2.0 mg/l gibberellic acid, solidified with 0.7% agar, pH 5.8) with 50 mg/l kanamycin and 500 mg/l carbenicillin for the elongation of buds. After 3-6 weeks, 1- to 2-cm-long elongated shoots are excised and planted on RM1 medium (MS basal medium supplemented with a hormone combination of 0.2 mg/l NAA and 0.1 mg/l IAA, solidified with 0.8% agar, pH 5.8) with 25 mg/l km and 200 mg/l carb for rooting. We tested four genotypes of pepper, and all presented a high differentiation efficiency (81.3% on average), elongation rate (61.5%) and rooting efficiency (89.5%). Polymerase chain reaction analysis results showed that 40.8% of the regenerated plantlets were transgenic plants.

Adenine↗

The role of shoot-localized processes in the mechanism of Zn efficiency in common bean.

Zn efficiency (ZE) is the ability of plants to maintain high yield under Zn-deficiency stress in the soil. Two bean ( Phaseolus vulgaris L.) genotypes that differed in ZE, Voyager (Zn-efficient) and Avanti (Zn-inefficient), were used for this investigation. Plants were grown under controlled-environment conditions in chelate-buffered nutrient solution where Zn(2+) activities were controlled at low (0.1 pM) or sufficient (150 pM) levels. To investigate the relative contribution of the root versus the shoot to ZE, observations of Zn-deficiency symptoms in reciprocal grafts of the two genotypes were made. After growth under low-Zn conditions, plants of nongrafted Avanti, self-grafted Avanti and reciprocal grafts that had the Avanti shoot scion exhibited Zn-deficiency symptoms. However nongrafted and self-grafted Voyager, as well as reciprocal grafts with the Voyager shoot scion, were healthy with no visible Zn-deficiency symptoms under the same growth conditions. More detailed investigations into putative shoot-localized ZE mechanisms involved determinations of leaf biomass production and Zn accumulation, measurements of subcellular Zn compartmentation, activities of two Zn-requiring enzymes, carbonic anhydrase and Cu/Zn-dependent superoxide dismutase (Co/ZnSOD), as well as the non-Zn-requiring enzyme nitrate reductase. There were no differences in shoot tissue Zn concentrations between the Zn-inefficient and Zn-efficient genotypes grown under the low-Zn conditions where differences in ZE were exhibited. Shoot Zn compartmentation was investigated using radiotracer ((65)Zn) efflux analysis and suggested that the Zn-efficient genotype maintains higher cytoplasmic Zn concentrations and less Zn in the leaf-cell vacuole, compared to leaves from the Zn-inefficient genotype under Zn deficiency. Analysis of Zn-requiring enzymes in bean leaves revealed that the Zn-efficient genotype maintains significantly higher levels of carbonic anhydrase and Cu/ZnSOD activity under Zn deficiency. While these data are not sufficient to allow us to determine the specific mechanisms underlying ZE, they certainly point to the shoot as a key site where ZE mechanisms are functioning, and could involve processes associated with Zn compartmentation and biochemical Zn utilization.

Adaptation, Physiological↗

Extra thin alginate film: an efficient technique for protoplast culture.

This paper reports an efficient protoplast culture technique, the "extra thin alginate film" technique. The development of this improved method of protoplast culture was an outcome of an assessment of the efficiency and shortcomings of various protoplast culture techniques. The efficiency of this technique was evaluated with two model plant systems, viz., Nicotiana tabacum and Lotus corniculatus, and a comparison was made with the "thin alginate layer" technique, another efficient protoplast culture system. Results indicate that the culture technique with extra thin alginate film is as efficient as the technique with thin alginate layer, with many additional advantages. The present innovation overcomes most of the limitations of protoplast culture techniques described so far and can now be applied to a wide variety of crops to check its general applicability.

Alginates↗

The efficiency of treatment strategies of general practitioners. A Malmquist index approach.

It is widely recognized that general practitioners (GPs) play a key role in determining the use of resources for ambulatory care. In addition to the GPs' working hours, these resources consist of the work of specialists and that of hospital physicians treating the GPs' referrals and the cost of medication and other measures induced by the GP. Different systems of remuneration differ in their power to lead to efficient service provision. This contribution provides empirical evidence on the efficiency of service provision by Austrian GPs. The analysis is based on data for some 600 GPs. The data comprise sufficient information to assess the GPs' efficiency with regard to the way they manage their cases. Data Envelopment Analysis, a nonparametric technique, is used to estimate the production frontier. The results suggest that almost one-half of the GPs in the sample have a relative efficiency of 0.8 or less. A Malmquist decomposition of the productivity change reveals a decline in productivity. This is due to a pronounced negative shift of the frontier whereas individual efficiency rises against the weaker benchmark of the new frontier.

Journal Article↗

Efficient mechanical disruption of Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii by a new high-pressure homogenizer and recovery of intracellular aminotransferase activity.

Microbiological studies often involve bacterial cell fractionation, which is known to be difficult for Gram-positive as compared to Gram-negative bacteria. Our purpose was to test the breaking efficiency of a new high-pressure pilot homogenizer for three Gram-positive species involved in dairy technology and to assess the activity of an intracellular aminotransferase. Varied pressures (50, 100 and 200 MPa) were applied to concentrated bacterial suspensions (1.2 mg dry weight/ml) of Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii. Breaking efficiency was estimated by decreases in optical density at 650 nm, cellular dry weight and viability. The proteins released were quantified and the residual intracellular aminotransferase activity was estimated using leucine as substrate. One run at 50 MPa was sufficient to break 80% of lactobacilli cells whereas 200 MPa were required for the same efficiency for L. lactis and P. freudenreichii. Whatever the pressure, leucine aminotransferase activity was recovered in the supernatant after cell breaking. This new high-pressure pilot homogenizer can allow rapid (20 s/run), easy, continuous and highly efficient cell breaking for intracellular enzyme recovery or other purposes. As the species tested were not phylogenetically related, and had different morphologies and cell wall compositions, we conclude that most Gram-positive bacteria may be broken efficiently by this new device.

Bacteriological Techniques↗

Methods for understanding super-efficient data envelopment analysis results with an application to hospital inpatient surgery.

We compare two techniques for increasing the transparency and face validity of Data Envelopment Analysis (DEA) results for managers at a single decision-making unit: multifactor efficiency (MFE) and non-radial super-efficiency (NRSE). Both methods incorporate the slack values from the super-efficient DEA model to provide a more robust performance measure than radial super-efficiency scores. MFE and NRSE are equivalent for unique optimal solutions and a single output. MFE incorporates the slack values from multiple output variables, whereas NRSE does not. MFE can be more transparent to managers since it involves no additional optimization steps beyond the DEA, whereas NRSE requires several. We compare results for operating room managers at an Iowa hospital evaluating its growth potential for multiple surgical specialties. In addition, we address the problem of upward bias of the slack values of the super-efficient DEA model.

Benchmarking↗

Strain differences in superovulatory response, embryo development and efficiency of transgenic rat production.

The differences between rat strains in superovulation response, in vitro and in vivo development of preimplantation embryos and overall transgenic efficiency was studied. The protocols for induction of superovulation using single injections of pregnant mare's serum gonadotropin (PMSG) or minipumps with follicle stimulating hormone (FSH) were compared in Lewis (LEW), Wistar-Kyoto (WKY), and stroke-prone spontaneously hypertensive rats (SHRSP) or Sprague-Dawley (SD) and Wistar rats as representative inbred or outbred strains, respectively. The percentage of mated animals with positive superovulatory response was similar in all strains (60.0-100%). The mean number of ova per donor was not dependent on the kind of hormonal treatment used within each rat strain. In general, females from outbred SD and Wistar rats were more responsive to hormonal treatments than animals from inbred rat strains. In addition, SD female rats produced a significantly higher number of embryos per female in response to PMSG-treatment compared to all other strains. Between the inbred strains, SHRSP was the most effective for superovulation. In vitro development of intact zygotes to the blastocyst stage was not different between SD, Wistar and SHRSP rats. In contrast, in vitro development of WKY zygotes was significantly less efficient than in other strains. However, 2-cell stage embryos in vivo produced from SD, SD x Wistar and WKY animals showed no difference in competence to develop to blastocyst stage in vitro. The proportion of offspring developing after oviduct transfer of intact zygotes was similar in all strains (44.0-56.4%) with the exception of WKY rats (35.9%). We also compared the survival rate after injection, ability of manipulated zygotes to develop to term and overall transgenic efficiency in various rat strains. SD and SHRSP zygotes survived after microinjection better than the WKY and Lewis zygotes. No differences were found in the efficiency of transgene integration per newborn in different strains ranging from 5.7 to 16.7%. The results of this study demonstrate that different rat strains have varying responses to superovulation, sensitivity to microinjection, capability to develop in vitro until blastocyst stage or in vivo to term after transfer to foster mothers. Despite these differences all studied strains can be used for efficient transgenic rat production.

Animals↗

Effect of accessory pathway location on the efficiency of heating during radiofrequency catheter ablation.

During radiofrequency catheter ablation of accessory pathways there is a poor correlation between applied power and temperature at target sites for catheter ablation. This study was designed to examine the relation between power and temperature during radiofrequency catheter ablation in patients with accessory pathways and to identify the factors that affect the efficiency of heating, defined as the ratio of applied power and temperature. Twenty-nine patients underwent radiofrequency catheter ablation of an accessory pathway. Among 257 energy applications, 108 were applied for ablation of a right-sided accessory pathway, 105 for a left-sided accessory pathway, and 44 for a posteroseptal accessory pathway. During each application of radiofrequency energy, temperature was continually monitored by use of an ablation catheter with a thermistor embedded in the tip of the distal electrode. During some applications of energy, fluctuations in temperature were observed. The average power, impedance, temperature, and efficiency of heating for all applications of radiofrequency energy was 37 +/- 11 W, 100 +/- 9 ohms, 53 +/- 9 degrees C, and 1.7 +/- 0.8 degrees C/W (range 0.9 degrees to 6.6 degrees C/W), respectively. The efficiency of heating varied by location (p < 0.0001), with the greatest efficiency of heating for posteroseptal energy applications (2.3 +/- 1.2 degrees C/W, which were significantly greater than for left-sided (1.8 +/- 0.8 degrees C/W; p < 0.01) or right-sided (1.2 +/- 0.4 degrees C/W; p < 0.0001) applications. Phasic fluctuation in temperature was observed during 127 (49%) energy applications, and the efficiency of heating was 1.5 +/- 0.7 degrees C/W.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗