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PubMed1998

UGA codon position affects the efficiency of selenocysteine incorporation into glutathione peroxidase-1.

A UGA codon and a selenocysteine insertion sequence in the 3'-untranslated region are the only established mRNA elements necessary for selenocysteine (Sec or U) incorporation during translation. These two elements, however, do not universally confer efficient Sec incorporation. The objective of this study was to systematically examine the effect of UGA codon position on efficiency of Sec insertion. In a glutathione peroxidase-1 (F-GPX1) expression vector, the UGA at the native position (U47) was mutated to a cysteine codon, and codons for Ser-7, Ser-12, Ser-18, Ser-29, Ser-45, Ser-93, Cys-154, Val-172, Ser-178, and Ser-195 were individually mutated to UGA and transiently expressed in COS-7 cells. 75Se incorporation at the 11 positions was 31, 72, 54, 105, 90, 100, 146, 135, 13, 11, and 43%, respectively, of 75Se incorporation at U47, suggesting that Sec is more efficiently incorporated at UGA codons positioned in the middle of the coding region rather than close to the 5' or 3' ends. Ribonuclease protection showed that these differences were not due to differences in mRNA level. When the green fluorescence protein (GFP) coding region was placed in-frame at the 5' or 3' ends of the coding region in F-GPX1 to produce chimeric 50-51-kDa GFP/GPX1 proteins, Sec incorporation at UGA codons, formerly close to the 5' or 3' ends, was increased to levels comparable to the UGA at U47. Insertion of GFP after the UAA-stop was just as effective in increasing Sec insertion efficiency as GFP inserted before the stop. These studies used a recombinant expression model that incorporated Sec at non-native UGA codons at rates equal to those of endogenous glutathione peroxidase-1 and showed that the efficiency of Sec incorporation can be modulated by UGA position; Sec incorporation at high efficiency appears to require that the UGA be >21 nucleotides from the AUG-start and >204 nucleotides from the selenocysteine insertion sequence element.

3' Untranslated Regions↗

PubMed2004

Amino acid requirement of growing pigs depending on amino acid efficiency and level of protein deposition. 2nd communication: threonine.

This study was conducted to evaluate the variability of the efficiency of threonine in different feed proteins for growing pigs. This information is of importance for actual conclusions about threonine requirement within the exponential N-utilization model (Liebert and Gebhardt, 1986) used in our investigations. Wheat (as basal protein), high-protein soybean meal, low-protein soybean meal, rapeseed meal, field bean (Vicia faba), peas (Pisum sativum), corn gluten meal and soybean protein concentrate were used as protein sources. Fifty-six growing barrows (40-65 kg BW) of the genotype Piétrain x (Duroc x Landrace) were randomly allotted to eight N-balance experiments (n = 7). Diets were formulated with two main ingredients (wheat + one feed protein) with threonine as the first limiting amino acid in the mixture which was partly supplemented with crystalline amino acids. Based on N-balance data, the efficiency of threonine was determined in protein mixtures and individual feed proteins. Threonine requirement was calculated depending on efficiency of threonine and level of daily protein deposition. The results from the present studies indicate that the efficiency of threonine in different feed proteins varied in a wide range. Consequently, this factor has to be taken into account for requirement calculations. The threonine requirement depending on daily protein deposition (130, 145 and 160 g) and the efficiency of threonine according to different reference units (g/BW(kg)(-0.67)/d, g/d and % of threonine in the diet) were calculated. The threonine requirement of growing barrows (50 kg BW) corresponding to an average threonine efficiency was 8.52, 9.92 and 11.61 g/d for a daily protein deposition of 130, 145 and 160 g, respectively. The results for a daily protein deposition of 145 or 160 g are in agreement with actual studies and recommendations for threonine supply.

Animal Feed↗

PubMed1989

Stair stepping efficiency of mentally handicapped and nonhandicapped adult females.

Net efficiency for stair stepping was compared between 15 mentally handicapped (MH) and 15 nonhandicapped (NH) women of comparable chronological age. Efficiency was computed as the percentage of the energy output divided by the energy expenditure. Energy expenditure was assessed by the performance of subjects on a double stair stepping apparatus, stepping at three work rates (14, 17, and 19 asc/min), repeated on four test days. Open-circuit spirometric techniques were used to measure oxygen uptake. The three-way (Group x Stepping Rates x Days) repeated measures ANOVA indicated that: (1) NH women stepped more efficiently than MH women (p less than 0.01). Mean stepping efficiency was 17.1% for the NH group, and 15.6% for the MH group; (2) MH women did not show improvement in stepping efficiency over the four days, although there was a 20% increase in the number of MH subjects capable of completing the fastest stepping rate over the four days; and (3) MH subjects were more efficient at 17 and 19 asc/min than at 14 asc/min.

Adult↗

PubMed1991

Gross efficiency responses to exercise conditioning in adult males of various ages.

This study investigated gross efficiency changes in a group of 60 adult males (mean age 39.2 +/- 1.2 years) resulting from endurance training and age-related responses to such training in sub-groups (each n = 20) of younger (30.7 +/- 0.7 years), intermediate (38.3 +/- 0.5 years) and older (48.6 +/- 1.1 years) subjects. Gross efficiency (%) was calculated from work output, oxygen consumption and RER energy equivalents following 10 min standard cycle ergometry exercise at 100 W and 50 rev min-1. Measurements were made at pre-, mid- and post-8 months of training, which involved progressive walking/jogging activities designed to enhance endurance capacity. In the total group, VO2 decreased pre- to post-training from 2.15 +/- 0.02 to 1.93 +/- 0.01 1 min-1 (P less than 0.01). In the sub-groups, both the younger and older subjects showed a significantly reduced VO2, from 2.17 +/- 0.01 to 1.98 +/- 0.04 1 min-1 and 2.05 +/- 0.08 to 1.86 +/- 0.03 1 min-1 respectively (P less than 0.05), but no significant changes were noted at mid-training. In the intermediate age subjects, while there were trends towards a reduced VO2, none was significant. The ANOVA revealed increased mean gross efficiency in the total group from pre- (14.3 +/- 0.1%) to post- (15.5 +/- 0.2%) (P less than 0.05) but not at mid-training (14.8 +/- 0.2%). While similar trends were observed in the sub-groups, gross efficiency increases were not significant, although changes in gross efficiency were reflected in VO2. The findings suggest that during standardized exercise, oxygen cost may be reduced and gross efficiency increased in adult males following endurance training and that such changes may take place over a variety of age ranges.

Adult↗

PubMed2005

Effect of additives on encapsulation efficiency, stability and bioactivity of entrapped lysozyme from biodegradable polymer particles.

Low encapsulation efficiency, incomplete and erratic release profiles are the most common features of controlled released protein delivery systems employing biodegradable polymers. In the present study, lysozyme as a model protein was encapsulated in biodegradable microspheres using solvent evaporation method and the effect of amphiphilic stabilizer, a basic salt and a lyoprotectant on microparticle formulation was evaluated. Incorporation rat serum albumin (RSA) in the internal aqueous phase during emulsion increased the encapsulation efficiency of lysozyme and maintained the bioactivity. Use of NaHCO3 improved the encapsulation efficiency of lysozyme from 15-94%, but at the cost of reduced in vitro release characteristics. Incorporation of both RSA and NaHCO3 improved the bioactivity of lysozyme and decreased burst release of the protein from the polymer particle, but reduced the encapsulation efficiency from 90-70%. Addition of sucrose in the internal aqueous phase lowered the encapsulation efficiency which was restored by its addition in the external aqueous phase. Maintenance of internal aqueous phase pH close to the iso-electric point of the protein and osmotic balance between the internal aqueous phase and the external aqueous phase during solvent evaporation method helped in better encapsulation of the protein drug. In vitro release of the lysozyme correlated with the effect of different excipients on entrapment in polymer matrix. Entrapment efficiency as high as 76%, low burst effect and high bioactivity of the entrapped lysozyme was observed from the polymer particles. Use of RSA, sucrose and NaHCO3 helped in a co-operative way towards the formulation of particles entrapping bioactive lysozyme.

Absorption↗

PubMed2005

Effects of divalent cations on drug encapsulation efficiency of deacylated gellan gum.

The objective of this research was to evaluate the effects of various divalent cations on the encapsulation efficiency of gellan gum and to probe the underlying mechanisms responsible for drug-loading efficiency. Spherical beads containing azathioprine were prepared from deacetylated gellan gum by ionotropic gelation method. One molar solution of various divalent chlorides (MgCl(2), BaCl(2), CaCl(2), CuCl(2) and ZnCl(2)) and two additional concentrations of CaCl(2) (2.5 M and 5.0 M) were used as ionotropic media. Drug solubility was also determined in these ionotropic media and statistically evaluated using ANOVA. Solubility in various divalent chloride solutions (1.0 M) suggests that azathioprine forms complex with Ca(2+), Zn(2+) and Cu(2+), while there might be a formation of poorly water-soluble chelates with Mg(2+) and Ba(2+) as solubility in these media were less than in deionized water. The encapsulation efficiency of gellan gum was much higher in the presence of transition elements (Cu(2+) and Zn(2+)) when compared to alkaline earth metal ions (Ca(2+), Mg(2+) and Ba(2+)). Higher concentrations of Ca(2+) decreased the encapsulation efficiency of gellan gum in a nearly proportional manner. The correlation between encapsulation efficiency and pH of the ionotropic media was negative and significant (r=-0.9574, p<0.05), although the solubility of azathioprine seems to be independent of the pH of the ionotropic medium. Overall, the results suggest that drug encapsulation efficiency of deacetylated gellan gum is largely affected by the concentration and nature of various divalent cations (e.g. atomic number, valency or electro-positivity, co-ordination property, etc.) and pH of the ionotropic medium.

Antimetabolites↗

PubMed2000

Experimental evaluation of a mathematical model for predicting transfer efficiency of a high volume-low pressure air spray gun.

The transfer efficiency of a spray-painting gun is defined as the amount of coating applied to the workpiece divided by the amount sprayed. Characterizing this transfer process allows for accurate estimation of the overspray generation rate, which is important for determining a spray painter's exposure to airborne contaminants. This study presents an experimental evaluation of a mathematical model for predicting the transfer efficiency of a high volume-low pressure spray gun. The effects of gun-to-surface distance and nozzle pressure on the agreement between the transfer efficiency measurement and prediction were examined. Wind tunnel studies and non-volatile vacuum pump oil in place of commercial paint were used to determine transfer efficiency at nine gun-to-surface distances and four nozzle pressure levels. The mathematical model successfully predicts transfer efficiency within the uncertainty limits. The least squares regression between measured and predicted transfer efficiency has a slope of 0.83 and an intercept of 0.12 (R2 = 0.98). Two correction factors were determined to improve the mathematical model. At higher nozzle pressure settings, 6.5 psig and 5.5 psig, the correction factor is a function of both gun-to-surface distance and nozzle pressure level. At lower nozzle pressures, 4 psig and 2.75 psig, gun-to-surface distance slightly influences the correction factor, while nozzle pressure has no discernible effect.

Air Pollutants, Occupational↗

PubMed2006

Modeling of indoor air treatment of polychlorinated dibenzo-p-dioxins and dibenzofurans using high-efficiency particulate air-carbon filtration.

A high-efficiency particulate air (HEPA)-carbon filtration system was developed by the Access Business Group, LLC, to reduce the indoor levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). The HEPA filter removes the particle-bound PCDD/Fs, and the carbon filter removes the gaseous fraction. Because of the toxicity of PCDD/Fs, it is very difficult to handle them in the laboratory. In this study, mathematical modeling was performed to evaluate the performance of the HEPA-carbon filtration system for PCDD/Fs removal and to optimize its design and operation. The model was calibrated with experimental data conducted with toluene in a sealed room. Model simulations with four selected congeners demonstrated that it takes approximately 1 hr for the indoor air treatment system to reach the maximum removal efficiency and that the carbon air filter has a life time of 10(7) yr for dioxin removal. Given a zero emission from the HEPA filter, the overall removal efficiency is 78.7% for 2,3,7,8-tetrachloro dibenzo-p-dioxins, 89.8% for octa-chlorodibenzodioxin, 78% for tetra-chlorodibenzofuran, and 89.8% for octa-chlorodibenzofuran. The larger the mass emission from the HEPA filter, the lower the overall removal efficiency, and the larger the ratio of the filter flow rate (Q(f)) to the room flow rate (Q), the higher the overall removal efficiency. When the ratio of Q(f)/Q is 15, an overall removal efficiency of 90% can be reached for all four of the selected compounds. The removal of the four selected compounds does not change as the relative humidity increases < or = 90%.

Adsorption↗

PubMed2002

Rationale and cost-efficiency compared for urine or saliva testing and behavioural inquiry among UK offender populations: injectors, arrestees and prisoners.

BACKGROUND: Costs, methodology and efficiency at obtaining behavioural information and biological sample to be tested for blood-borne viruses or illegal drugs are compared for UK surveys of offender populations: injectors in the community or attending drug agencies, arrestees and prisoners. METHODS: Arrestee surveys use extensive behavioural interview + urine sample and measure a key performance indicator in UK's Drugs Strategy. They have low efficiency (urine sample for drugs testing available from under 60% of eligible arrestees) at high cost (pound sterling 110-190 or 350 per urine sample and at least pound sterling 500-800 per injector). Random mandatory drugs testing of prisoners has the highest efficiency (because refusals are punishable), but the cost is high (pound sterling 110-120 per urine sample and pound sterling 300-500 per injector) and behavioural data are lacking. Prisoner surveys use self-completion questionnaire + saliva sample. They guarantee demonstrable anonymity in estimating the prevalence of blood-borne viruses and prisoners' associated risk behaviours, have high efficiency (saliva sample from over 80% of inmates) at low cost (pound sterling 30 per saliva and pound sterling 70-110 per injector), but behavioural data are limited to risk factors for blood-borne viruses. Low cost also characterises comprehensive interview + saliva sample from injectors in the community (pound sterling 90 per saliva sample, all from injectors) but efficiency cannot be assessed because the sampling frame of eligible injectors is not known. Voluntary unlinked anonymous surveys of injectors at drug agencies use self-completion questionnaire + saliva sample to be tested for blood-borne viruses. They are the least costly at pound sterling 43 per injector and moderately efficient with two-thirds volunteer rate by eligible injectors. DISCUSSION: For scientific added-value, we recommend co-location of survey types geographically and temporally; a common core set of behavioural questions; saliva sample as well as (and, eventually, instead of) the less acceptable urine sample. Survey methodologies for measuring key performance indicators should stand up to scrutiny in terms of openness, design credentials, statistical power, and costs. We examine how participation bias or inadequate survey size can compromise the effective monitoring of a key performance indicator.

Cost-Benefit Analysis↗

PubMed1998

Performance of N95 respirators: filtration efficiency for airborne microbial and inert particles.

In 1995 the National Institute for Occupational Safety and Health issued new regulations for nonpowered particulate respirators (42 CFR Part 84). A new filter certification system also was created. Among the new particulate respirators that have entered the market, the N95 respirator is the most commonly used in industrial and health care environments. The filtration efficiencies of unloaded N95 particulate respirators have been compared with those of dust/mist (DM) and dust/fume/mist (DFM) respirators certified under the former regulations (30 CFR Part 11). Through laboratory tests with NaCl certification aerosols and measurements with particle-size spectrometers, N95 respirators were found to have higher filtration efficiencies than DM and DFM respirators and noncertified surgical masks. N95 respirators made by different companies were found to have different filtration efficiencies for the most penetrating particle size (0.1 to 0.3 micron), but all were at least 95% efficient at that size for NaCl particles. Above the most penetrating particle size the filtration efficiency increases with size; it reaches approximately 99.5% or higher at about 0.75 micron. Tests with bacteria of size and shape similar to Mycobacterium tuberculosis also showed filtration efficiencies of 99.5% or higher. Experimental data were used to calculate the aerosol mass concentrations inside the respirator when worn in representative work environments. The penetrated mass fractions, in the absence of face leakage, ranged from 0.02% for large particle distributions to 1.8% for submicrometer-size welding fumes. Thus, N95 respirators provide excellent protection against airborne particles when there is a good face seal.

Aerosols↗

PubMed2006

House dust collection efficiency of the high volume small surface sampler on worn carpets.

The High Volume Small Surface Sampler (HVS3) is a dust-sampling vacuum that allows for set airflow and back pressure during sampling, increasing precision. Total dust collection efficiency of the HVS3 has been evaluated only on new carpets-not worn carpets. We performed a factorial study to assess the impact of carpet wear, dust deposition level, carpet type, and relative humidity during sampling on HVS3 collection efficiency. House dust was aerosolized in a 1-m3 exposure chamber and allowed to settle on test carpets and reference filters. Dust was embedded into the carpets and later extracted with the HVS3 under controlled environmental conditions according to established protocols. Overall collection efficiency was high, 88.3%. Collection efficiency was significantly higher at low relative humidity levels (30%) relative to high (75%) (p = < 0.0001), though differentially between cut-pile and closed-loop carpets. Collection efficiency of carpets with high wear was significantly lower than those with midlevel wear (p = 0.01). These results demonstrate that the design of the HVS3 partially corrects for differences in dust load and carpet type. However, collection efficiency of the HVS3 is affected by high levels of carpet wear and ambient humidity during sampling.

Analysis of Variance↗

PubMedDate not supplied

Physiology and behavior influence lactation efficiency in northern elephant seals (Mirounga angustirostris).

The efficiency with which mothers convert acquired energy into milk is a key determinant of the magnitude of parental investment in mammals; however, the mechanisms underlying lactation efficiency are poorly understood. Investigations on northern elephant seals have shown lactation efficiency, measured as the proportion of total energy expenditure that goes to the pup as milk, increases with age. In a cross-sectional study the physiological and behavioral determinants of lactation efficiency were investigated in eight young and seven prime (older) elephant seals by conducting behavioral observations and collecting milk, blood, and tissue on days 3 and 22 of lactation. Milk composition, circulating fatty acid and triglyceride concentrations, and mammary and blubber lipoprotein lipase activity were determined. Prime females had significantly greater percent milk fat and circulating fatty acids on day 3 than did young females, but these differences disappeared by day 22. The ability for prime females to produce higher-energy milk early in lactation may allow them to increase lactation efficiency by increasing the rate of energy transfer. In addition, prime females spent significantly more time resting. A combination of reduced activity and more rapid energy delivery likely explains the increase in lactation efficiency with age found in a previous study.

Age Factors↗

PubMed2006

Efficiency of human immunodeficiency virus transmission through injections and other medical procedures: evidence, estimates, and unfinished business.

OBJECTIVE: To estimate the transmission efficiency of human immunodeficiency virus (HIV) through medical injections and other invasive procedures. DESIGN: We searched our own files and Medline (from 1966-2004, using the keywords ["iatrogenic" or "nosocomial" or "injections"] and "HIV") for reports of iatrogenic outbreaks worldwide, except outbreaks traced to receipt of contaminated blood or blood products. We also analyzed information from a case-control study of percutaneous exposures to healthcare workers. SETTING: Worldwide healthcare settings. EVENTS: We identified 8 iatrogenic outbreaks that met our study criteria; published information from 4 outbreaks was sufficient to estimate transmission efficiency. RESULTS: From the 4 documented iatrogenic outbreaks, we estimated that 1 iatrogenic infection occurred after 8-52 procedures involving HIV-infected persons. Although only 0.3% of healthcare workers seroconvert after percutaneous exposure, a case-control study reported that deep injuries and other risk factors collectively increased seroconversion risk by as much as 50 times. Laboratory investigations demonstrate HIV survival through time and various rinsing regimens. We estimate that the transmission efficiency in medical settings with no or grossly insufficient efforts to clean equipment ranges from 0.5% to 3% for lower risk procedures (eg, intramuscular injections) and from 10% to 20% or more for high-risk procedures. Efforts to clean equipment, short of sterilization, may cut the transmission efficiency by 0%-100%. Procedures that contaminate multidose vials may accelerate transmission efficiency. CONCLUSION: To achieve better estimates of the transmission efficiency for a range of medical procedures and settings, investigations of iatrogenic outbreaks should be accorded high priority.

Adult↗

PubMedDate not supplied

Apparent absorption efficiencies of nectar sugars in the Cape sugarbird, with a comparison of methods.

Nectarivore sugar preferences and nectar composition in the Cape Floristic Kingdom (southern Africa) differ from trends reported for analogous systems in America and Europe in that sugarbirds and sunbirds show no aversion to sucrose, which is the dominant nectar sugar in many of their food plants. To elucidate the physiological bases (if any) of nectarivore sugar preferences, we determined apparent sugar absorption efficiencies in a passerine endemic to this region, the Cape sugarbird Promerops cafer. Apparent absorption efficiencies for the three major nectar sugars, sucrose, glucose, and fructose, were extremely high (> 99%), as in other specialized avian nectarivores. Xylose, a pentose sugar recently reported in the nectar of some Proteaceae, was absorbed and/or metabolized inefficiently, with a mean of 47.1% of ingested sugar recovered in cloacal fluid. We did not measure the proportions of xylose that were absorbed and/or metabolized. We also compared three methods of estimating absorption efficiency: (1) measurements of total sugar in cloacal fluid with refractometry, without correction for differences between volumes of ingesta and excreta; (2) the same measurements combined with correction for volume differences; and (3) HPLC analyses quantifying individual sugars in cloacal fluid, with correction for volume differences. Refractometry has been frequently used in previous studies. For all sugars except xylose, method 1 yielded results similar to those obtained with method 2, but the convergence was artifactual, and we do not recommend use of this method. Apparent absorption efficiencies calculated with method 2 underestimated true absorption efficiency, because refractometry measures nonsugar solutes, but this error is biologically significant only when efficiencies are low.

Absorption↗

PubMed2000

Highly efficient adenovirus-mediated gene transfer to cardiac myocytes after single-pass coronary delivery.

Efficient and homogeneous gene transfer to cardiac myocytes is a major target in myocardial gene therapy. The aim of this study was to determine the conditions permitting efficient, homogeneous, adenovirus-mediated gene transfer to cardiac myocytes, with a view to application during coronary artery catheterization. Gene transfer to adult rat ventricular myocytes was conducted using type 5 adenoviruses carrying the lacZ reporter gene. Adenovirus delivery via coronary arteries was performed on isolated perfused rat hearts, and gene transfer efficiency was analyzed on whole ventricles, freshly isolated myocytes, and cultured myocytes. Single-pass delivery of 1 X 10(9) PFU associated with 1 min of no-flow yielded only 1 +/- 0.5% of positive myocytes. Pretreatment by histamine perfusion (10(-5) M final concentration) increased this value to 30 +/- 9% (p < 0.001), and pretreatment by Ca2+-free buffer perfusion increased it to 67 +/- 8% (p < 0.001). Combination of the two pretreatments had no additional effect. Increasing the viral dose to 3 X 10(9) PFU increased transfection efficiency only in permeabilized vessels. The 1-min no-flow period after adenovirus delivery was crucial for efficient gene transfer: despite histamine pretreatment, only 2 +/- 1% positive myocytes were observed without flow interruption (p < 0.05 versus 1 min of no-flow). Gene transfer was shown to occur in situ during cardiac perfusion, rather than during heart digestion or myocyte isolation. This study shows that highly efficient adenovirus-mediated gene transfer to cardiac myocytes in situ can be achieved by single-pass intracoronary vector delivery, provided that vascular permeability is first increased and coronary flow is briefly interrupted.

Adenoviridae↗

PubMed2003

Efficient trans-splicing in the retina expands the utility of adeno-associated virus as a vector for gene therapy.

Recombinant vectors based on adeno-associated virus (AAV) can efficiently transduce many different cell types, including cells of the retina, resulting in stable gene expression. A major shortcoming of this vector is its small packaging capacity. A trans-splicing approach, which reconstitutes gene expression from two independent AAV vectors, can be used to overcome the vector's packaging limitations. The efficiency of this system to date has been disappointing, and therefore its utility for therapeutic application limited. We demonstrate here that efficiency and cellular specificity of trans-splicing is dependent on selection of the appropriate AAV serotype. Efficiency of transgene expression resulting from trans-splicing in skeletal muscle approaches that obtained when delivering the intact transgene when using AAV2 vectors packaged with AAV5 capsids (AAV2/5). This expands the potential of AAV vectors for retinal gene therapy. The use of AAV2/5 also increases the efficiency of trans-splicing in photoreceptors. Selection of the appropriate AAV serotype is likely to affect efficiency of trans-splicing in other organ systems as well.

Animals↗

PubMed1999

Quantitative analysis of the functionality and efficiency of three surgical dissection techniques: a time-motion analysis.

The increasing technological complexity of surgery demands objective evaluation of surgical techniques. In particular, alternatives for laparoscopic ligation, such as monopolar coagulation and the relatively new bipolar scissors combining dissection with coagulation, should be analyzed and compared. This study tests the efficacy of quantitative time-motion analysis in evaluating and comparing the functionality and efficiency of dissection and ligation techniques in a clinical setting. Standard dissection with ligation of vessels, bipolar scissors, and monopolar coagulation were consecutively applied to dissect 4 of the small bowel mesentery of pigs, in random order. All actions performed were recorded and analyzed, using a standard action list. The efficiency of each technique was expressed in mean dissection time and number of actions, and the safety in occurrence of complications and severity of microscopic damage. Time-motion analysis evaluated the efficiency objectively and reproducibly (ICC 0.98). Bipolar scissors were significantly more efficient (time 7 +/- 2 min, actions 129 +/- 33) than the standard technique (28 +/- 6, 771 +/- 185) and monopolar coagulation (14 +/- 5, 368 +/- 32) (p < 0.01). Furthermore, bipolar coagulation needed significantly less recoagulation of an oozing vessel (0.5% of the total dissected vessels) than did monopolar coagulation (10.4%), and the damaged zone was significantly smaller (p < 0.05). Significantly less time was spent waiting or exchanging instruments with bipolar scissors than with the standard technique (p < 0.05). This time-motion analysis objectively compared the efficiency and functionality of three surgical dissection techniques during clinical use. Bipolar scissors were more efficient than were both other techniques, and they coagulated vessels more safely than did monopolar coagulation.

Animals↗

PubMed1998

Increased transduction efficiency of primary hematopoietic cells by physical colocalization of retrovirus and target cells.

Efficient gene transfer into hematopoietic stem cells offers a number of potential therapeutic applications. However, the relatively low titer of retroviral supernatants and the requirement for cell division to ensure integration have meant that transduction efficiency has been low. We have modified a flowthrough approach to cell transduction and have been able consistently to increase gene transfer efficiency into human hematopoietic progenitor cells. We transduced CD34 cells with retroviral vectors encoding a truncated nerve growth factor receptor (NGFR) or neo. Retroviral supernatant was pulled through 0.2-micron polycarbonated membranes, followed by placement of cells on the filter. In the absence of cytokines, the transduction efficiency of CD34 cells with a NGFR vector was increased 3-11-fold over that obtained at an identical MOI in liquid culture to produce 11%-44% transduction. Furthermore, both Thy1+ and Thy1- subsets in a total CD34 population were transduced with similar efficiency, and transduction with a neo vector, as measured by G418 resistance in clonogenic assays, increased 1.5-5-fold. The mechanism by which gene transfer is improved may reflect colocalization of cells and retrovirus. Costaining of cells transduced on the filter with an NGFR retrovirus with both an NGFR antibody and a gp70 antibody that recognizes viral coat protein revealed high-level coexpression. The levels of in vitro gene transfer we obtain are equivalent to those observed when CD34 cells are cocultured in liquid culture with cytokines. However, culture with cytokines may commit CD34 cells to differentiation and has produced disappointingly low levels of subsequent in vivo gene transfer. Gene marking studies using distinguishable retroviral vectors will provide a means of learning whether the effects of flowthrough transduction genuinely enhance the efficiency of gene transfer to human marrow-repopulating cells.

Antigens, CD↗