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The effect of methanol washing of plasticized polyvinyl chloride on biomaterial-contact-mediated CD11b (mac-1) expression in a rat recirculation model.

Our objective was to assess whether using a methanol wash to reduce the level of plasticizer present on the surface of medical-grade polyvinyl chloride (PVC) has a moderating effect on the expression of CD11b (mac-1) on neutrophils in rats undergoing recirculation. The study was carried out on 3 groups of 10 adult male Sprague-Dawley rats weighing between 350 and 450 g. In the 2 test groups, the animals were exposed to 48 cm(2) of di-(2-ethyl-hexyl)-phthalate (DEHP)-plasticized PVC in a parallel plate recirculating test cell through which blood was recirculated at 1.5 ml/min. In the first test group, the PVC was untreated; in the second test group, the PVC was washed in methanol to reduce the level of plasticizer on the surface. The test cell was connected to the right femoral circulation, and recirculation was established for a period of 60 min. Blood samples were taken at 0, 30, and 60 min for assessment of CD11b expression on neutrophils using flow cytometric analysis. In a third group of 10 control experiments, rats underwent the entire surgical procedure, but without recirculation through the test cell. There was statistically significant (p < 0.001) lower Cd11b expression on neutrophils in the blood of rats perfused through the cell containing methanol-washed PVC after 30 min and at 60 min. CD11b expression was significantly (p < 0.001) lower in the control group than in both test groups at both the 30 and 60 min time points and at the 60 min time point on comparison with the group where blood was perfused through methanol-washed PVC. These results demonstrate that the biomaterial-contact-mediated upregulation of CD11b may be significantly reduced by employing a methanol-washing technique on the plasticized PVC. Although this technique does not entirely eliminate the expression of CD11b on neutrophils, the difference is significant and suggests the role of the plasticizer in the development of this inappropriate inflammatory response.

Animals↗

Analysis of saline-washed red cells for transfusion to neonatal patients.

A protocol was developed under which washed red cells were used to provide small-volume transfusions for neonatal patients (under 4 months of age). Two pediatric units, each containing approximately 135 ml of red cells, were prepared from a freshly collected unit of group O Rh-negative whole blood. Units not more than 6 days old were washed with 1l of 0.9 percent NaCl in a blood cell processor. Aliquots of the washed red cells were drawn into labeled syringes for transfusion to neonatal patients. Data collected from 40 units of washed red cells prepared according to this protocol showed 83 percent red cell recovery, and satisfactory reduction in white cells, plasma proteins, extracellular potassium, red cell metabolic waste products, and anticoagulant. Over 5 years, a total of 2085 different neonatal patients received 7875 separate transfusions prepared from 1860 units of washed red cells. The clinical response was excellent, and no suspected reactions were reported.

Blood Proteins↗

[Automated quantitative image cytometry of bronchial washings in suspected lung cancer: comparison with cytology, histology and clinical diagnosis].

INTRODUCTION: Automated image cytometry represents a new method for the quantitative analysis of nuclear structure and DNA-content of exfoliative airway epithelial cells. In the present investigation, we examined the correlation between automated cytometry, conventional cytology and histopathology with the final diagnosis as the "gold standard". METHODS: In 142 patients (100 males and 42 females) with suspected lung cancer and 50 controls (COPD, asthma), bronchial washings (5-10 ml) were obtained during bronchoscopy before taking biopsies for cytological and/or histological examinations. The washings were collected in 20 ml Saccomanno's fixative and centrifuged (500 g, 15 min). The cell pellet was resuspended in Saccomanno's solution. Two specimens were stained according to Papanicolaou and another two using the Feulgen reaction with thionine. Image cytometry was performed by means of a special, trainable classifier for exfoliative cells of the respiratory tract, using the Cyto-Sacant (Oncometrics, Vancouver). RESULTS: In the patients with suspected lung cancer we found numerous abnormal nuclei in 97 samples, 36 samples contained normal cells only, and 9 samples were insufficient. In our control group there was no sample with abnormal nuclei, and all washings were evaluable. Compared to the final diagnosis of lung cancer, we found a sensitivity of 90% (92/102) and a specificity of 84% (26/31). For histology sensitivity was 91% (73/80) and specificity 100%, while we found a sensitivity of 92% (92/100) and specificity of 100% for cytology. For automated cytometry the positive predicted value was 95%, the negative predicted value 71%. CONCLUSIONS: In the investigation of patients with suspected lung cancer, automated image cytometry of bronchial washings is a sensitive and reliable method for the detection of malignant changes in the tracheobronchial mucosa. The automated procedure seems well suited not only for analysing bronchial washings, but also for a screening procedure.

Adult↗

Comparative efficacy of alternative hand-washing agents in reducing nosocomial infections in intensive care units.

BACKGROUND: Effective hand-washing can prevent nosocomial infections, particularly in high-risk areas of the hospital. There are few clinical studies of the efficacy of specific hand-cleansing agents in preventing the transmission of pathogens from health care workers to patients. METHODS: For eight months, we conducted a prospective multiple-crossover trial involving 1894 adult patients in three intensive care units (ICUs). In a given month, the ICU used a hand-washing system involving either chlorhexidine, a broad-spectrum antimicrobial agent, or 60 percent isopropyl alcohol with the optional use of a nonmedicated soap; in alternate months the other system was used. Rates of nosocomial infection and hand-washing compliance were monitored prospectively. RESULTS: When chlorhexidine was used, there were 152 nosocomial infections, as compared with 202 when the combination of alcohol and soap was used (adjusted incidence-density ratio [IDR], 0.73; 95 percent confidence interval, 0.59 to 0.90). The largest reduction with chlorhexidine was in gastrointestinal infections (IDR, 0.19; 95 percent confidence interval, 0.05 to 0.64). When chlorhexidine was available, the rates of nosocomial infection declined in each of the ICUs, and health care workers washed their hands more often than when alcohol and soap were used (relative risk, 1.28; 95 percent confidence interval, 1.02 to 1.60). The total volume of alcohol and soap used was 46 percent that of chlorhexidine (P less than 0.001). CONCLUSIONS: A hand-disinfection system using an antimicrobial agent (chlorhexidine) reduces the rate of nosocomial infections more effectively than one using alcohol and soap. The improvement may be explained at least in part by better compliance with hand-washing instructions when chlorhexidine was used.

1-Propanol↗

Long-term prognostic value of positive peritoneal washing in colon cancer.

BACKGROUND: We examined the potential prognostic value of free malignant cells in the peritoneal cavity of patients with colon adenocarcinoma. METHODS: In 1991 and 1992, 88 patients underwent peritoneal washing with cytologic analysis immediately before elective colon resection for adenocarcinoma with no evidence of peritoneal metastases. Peritoneal washing fluid was collected before abdominal exploration, centrifuged immediately, and stained with the May-Grünwald-Giemsa and Harris Shorr methods. Malignancy was defined as recommended by Papanicolaou. RESULTS: Free malignant cells in peritoneal fluid (positive peritoneal washing) were found in 25 of 88 (28%) patients (Dukes A, 0 of 11; Dukes B, 10 of 31; Dukes C, 11 of 37; Dukes D, 4 of 9). The positivity rate was 24 of 75 (32%) among patients with tumours involving the serous layer and 1 of 13 (8%) among the others (P = 0.0989). With a follow-up of 66.7 to 78.7 months the 5-year survival rates were 48% (95% confidence interval, 28.4-67.6) and 68% (95% CI, 56.2-79.8) among patients with positive and negative peritoneal washing, respectively (P = 0.09). Positive peritoneal washing was not significantly associated with survival in multivariate analysis. CONCLUSION: The presence of free malignant cells in the peritoneal cavity of patients with colon cancer provided no further prognostic information relative to the Dukes classification in this study. Nevertheless, further study is needed, particularly in a larger series of Dukes B patients, for whom a new prognostic factor would be useful for deciding adjuvant therapy.

Adenocarcinoma↗

Aged chlorophenol contaminated soil's integrated treatment by ozonation, soil washing and biological methods.

Traditionally soils contaminated by organic compounds have been treated by biological methods, but aged contaminated soils usually contain refractory and toxic compounds not any more responsive to biological treatment. By oxidation and soil washing the biodegradability and bioavailability of these otherwise recalcitrant compounds can be enhanced. The aim of the study was to optimize and integrate soil washing, ozonation (+ hydrogen peroxide) and biological treatment for the remediation of old saw mill soil contaminated by chlorophenols (CPs). The integration of different treatment train variables and the alternation of pHs (3, 7 and 10) and different ozone doses were studied in a laboratory scale. The soil was heavily contaminated aged soil (> 4000 mgCP kg(-1)), so more than 99% of removal should be attained in order to reach the Finnish guideline values. All the individual methods studied were able to degrade/transfer 25 - 95% of the CPs from the soil matrix. By biological treatment only 25% of the CPs were degraded. After soil washing, 40- 80% of the contaminants were transferred from the soil phase and by ozonation 35 - 95% decrease of CPs were achieved. All the methods studied successfully enhanced the biodegradability of the target compound. After biological treatment of ozonated and washed samples even 93 - 100% degradation of CPs was obtained. The most effective treatment was the combination of soil washing + ozonation (in pH 10) followed by biological treatment.

Biodegradation, Environmental↗

Volatilization of chemicals from drinking water to indoor air: the role of residential washing machines.

Previous research has indicated that residential washing machines are potential sources of pollution due to the associated use of chemicals found in consumer products, for example, ethanol in laundry detergent and chlorine in bleach. Washing machines may also emit hazardous air pollutants found in contaminated drinking water. To better understand the extent and impact of chemical emissions from tap water, 26 experiments were completed using a residential washing machine and a cocktail of chemical tracers representing a wide range of physicochemical properties. Variable operating conditions for these experiments included water temperature, amount of clothes present in the machine, water volume, and level of washwater agitation. Chemical stripping efficiencies and mass transfer coefficients were determined during each cycle (fill, wash, and rinse) of a normal washing machine event. Headspace ventilation rates were determined using an isobutylene tracer gas. Mass transfer rates were significantly influenced by operating parameters as exhibited by a wide range of chemical stripping efficiencies. Stripping efficiencies ranged from 0.74 to 36% for acetone, 8.2 to 99% for toluene, 10 to 99% for ethylbenzene, and 6.9 to 100% for cyclohexane.

Air Pollution, Indoor↗

A simple and rapid method of washing and drying micro-titre plates used in ELISA.

A simple device constructed out of perspex sheeting and capable of washing 96-well micro-titre plates used in ELISA is described. It consists of three parts: (a) the 'shower' - a tank with 96 stainless steel syringe needles through which washing fluid runs into the wells of the micro-titre plate, (b) a sump into which washing fluid is collected and (c) a sliding base plate allowing the accurate positioning of the micro-titre plate below the shower. The efficiency of this washing device was established by comparing the results from several specific ELISAs with tha obtained after washing duplicate plates by hand. The use of a specially constructed centrifuge to spin-dry plates is also described.

Enzyme-Linked Immunosorbent Assay↗

Methane production by methanogens following an aerobic washing procedure: simplifying methods for manipulation.

The recent discovery of methane in the martian atmosphere is arguably one of the most important discoveries in the field of astrobiology. One possible source of this methane could be a microorganism analogous to those on Earth in the domain Archaea known as methanogens. Methanogens are described as obligately anaerobic, and methods developed to work with methanogens typically include anaerobic media and buffers, gassing manifolds, and possibly anaerobic chambers. To determine if the time, effort, and supplies required to maintain anaerobic conditions are necessary to maintain viability, we compared anaerobically washed cells with cells that were washed in the presence of atmospheric oxygen. Anaerobic tubes were opened, and cultures were poured into plastic centrifuge tubes, centrifuged, and suspended in fresh buffer, all in the presence of atmospheric oxygen. Washed cells from both aerobic and anaerobic procedures were inoculated into methanogenic growth media under anaerobic conditions and incubated at temperatures conducive to growth for each methanogenic strain tested. Methane production was measured at time intervals using a gas chromatograph. In three strains, significant differences were not seen between aerobically and anaerobically washed cells. In one strain, there was significantly less methane production observed following aerobic washing at some time points; however, substantial methane production occurred following both procedures. Thus, it appears that aerobic manipulations for relatively short periods of time with at least a few species of methanogens may not lead to loss of viability. With the discovery of methane in the martian atmosphere, it is likely that there will be an increase in astrobiology-related methanogen research. The research reported here should simplify the methodology.

Aerobiosis↗

Studies on gonococcus infection. XVII. IgA1-cleaving protease in vaginal washings from women with gonorrhea.

Vaginal washings from women attending a veneral disease clinic were examined for the presence of protease that cleaved IgA subclass 1 (IgA1). In a crude assay, vaginal washings cleaved [125I]IgA1 in 19 of 25 specimens from individuals from whom Neisseria gonorrhoeae were cultivated. Forty-six specimens from 104 women whose cultures were negative for N. gonorrhoeae also cleaved [125I]IgA1. Vaginal washings from six of six women with culture-proven gonorrhea cleaved [125I]IgA1 into low-molecular-weight components identical to those produced by partially purified IgA1-specific protease from gonococci. The hydrolysis of [125I]IgA1 by vaginal washings from women whose cultures were negative for N. gonorrhoeae yielded cleavage products that resembled those of trypsin or alpha-chymotrypsin. These findings indicate that gonococci residing in the female genital tract produce IgA1-specific protease that can be detected in the vaginal washings of infected women.

Electrophoresis, Polyacrylamide Gel↗

Washing and cryoprotectant effects on frozen storage of spent hen surimi.

Mince of 98-wk-old spent hens was washed two times with 0.1% NaCl. Portions of unwashed and washed mince were mixed with a cryoprotectant (CP) composed of 4% sucrose, 4% sorbitol, and 0.2% Na-tripolyphosphate and were immediately frozen and stored at -20 C. Mince without CP was run as control. Textural properties of the stored mince and surimi were measured at 1-mo intervals for 6 mo, after being thawed at 4 C overnight, ground with 3% NaCl, and heated at 90 C for 15 min. For freeze-thaw stability study, minces were subjected to six freeze-thaw cycles. Each freeze-thaw cycle was carried out at 1 mo of storage. Textural quality parameters (gel strength, breaking strength, deformation, protein solubility, expressible moisture, cooking yield, folding test, drip-loss, and sensory scores) were decreased in both unwashed and washed mince, mostly during the early stages of storage. Washed mince showed significantly better textural properties than unwashed mince. Washing protected the gel quality of the hen mince from degradation during frozen storage. Cryoprotectants could not protect the gel strength or breaking strength, but deformation was slightly improved. Water-retention properties were protected, and folding test and sensory scores were well preserved in the mince with added CP. Cryoprotectants had a beneficial effect on frozen, stored spent hen surimi to protect the elasticity and cohesiveness of the gel.

Animals↗

Percoll gradient-centrifuged capacitated mouse sperm have increased fertilizing ability and higher contents of sulfogalactosylglycerolipid and docosahexaenoic acid-containing phosphatidylcholine compared to washed capacitated mouse sperm.

Although Percoll gradient centrifugation has been used routinely to prepare motile human sperm, its use in preparing motile mouse sperm has been limited. Here, we showed that Percoll gradient-centrifuged (PGC) capacitated mouse sperm had markedly higher fertilizing ability (sperm-zona pellucida [ZP] binding and in vitro fertilization) than washed capacitated mouse sperm. We also showed that the lipid profiles of PGC capacitated sperm and washed capacitated sperm differed significantly. The PGC sperm had much lower contents of cholesterol and phospholipids. This resulted in relative enrichment of male germ cell-specific sulfogalactosylglycerolipid (SGG), a ZP-binding ligand, in PGC capacitated sperm, and this would explain, in part, their increased ZP-binding ability compared with that of washed capacitated sperm. Analyses of phospholipid fatty acyl chains revealed that PGC capacitated sperm were enriched in phosphatidylcholine (PC) molecular species containing highly unsaturated fatty acids (HUFAs), with docosahexaenoic acid (DHA; C22: 6n-3) being the predominant HUFA (42% of total hydrocarbon chains of PC). In contrast, the level of PC-HUFAs comprising arachidonic acid (20:4n-6), docosapentaenoic acid (C22:5n-6), and DHA in washed capacitated sperm was only 27%. Having the highest unsaturation degree among all HUFAs in PC, DHA would enhance membrane fluidity to the uppermost. Therefore, membranes of PGC capacitated sperm would undergo fertilization-related fusion events at higher rates than washed capacitated sperm. These results suggested that PGC mouse sperm should be used in fertilization experiments and that SGG and DHA should be considered to be important biomarkers for sperm fertilizing ability.

Animals↗

Importance of wash riding in kayaking training and competition.

PURPOSE: The use of different wash-riding techniques is common during kayak training and competition. Changes in wash-riding positions could imply a different exercise intensity. The aim of this study, therefore, was to quantify the energy savings made when a kayaker is "wash riding." METHODS: Eight male international flat water kayakers, who performed a field test of 2000 m in each of the four wash-riding positions, head (H), right wave (RW), left wave (LW), and end position (V), were studied. The data investigated were: time, stroke rate, blood lactate (BL), heart rate (HR), and rate of perceived exertion (RPE). Under laboratory conditions kayakers performed the same intensity of exercise in a kayak ergometer, and HR, oxygen uptake (VO2), BL, mean power output (W), and RPE were measured. RESULTS: The results show significant differences (P < 0.05) among H, RW/LW, and V. The mean values for BL (P < 0.05) were 4.2, 2.0, 2.2, and 1.5 mmol.L-1, for H, RW, LW, and V, respectively. RPE also revealed differences, with values of 15, 12.6, 12.6, and 9.7 for H, RW, LW, and V, respectively. Mean power output gave values of 190.3 (H), 155.6 (RW and LW), and 129.5 (V) W. HR was different between H and V (172 and 151), while stroke rate was different among the parameters H, RW, and V (93.7, 88.8, and 87.6, respectively). The VO2 in the kayak ergometer test showed a difference between H and V (3.78 and 2.23 L.min-1). CONCLUSIONS: We conclude that "wash riding" involves a saving in energy cost of between 18% and 31.9%, depending on the position. This conclusion is of importance for the quantification and calibration of kayak training and competition.

Adult↗

Treponemicidal antibody measured by the "washed-killing" assay correlates with immunity in experimental rabbit syphilis.

BACKGROUND AND OBJECTIVES: The authors have previously shown that complement-dependent treponemicidal antibody measured by the "washed-killing" assay is directed exclusively against surface-exposed targets on Treponema pallidum, presumably the Treponema pallidum rare outer membrane proteins detected by freeze-fracture electron microscopy. GOAL OF THIS STUDY: Because immune mechanisms against Treponema pallidum rare outer membrane proteins are likely to be central to a protective host response, it was examined whether a relationship could be established between treponemicidal levels as measured by the "washed-killing" assay and host immunity in experimental syphilis. STUDY DESIGN: Three groups of Treponema pallidum-infected rabbits were treated curatively with penicillin at 9 days, 30 days, and 6 months post-infection to generate animals with varying degrees of immunity to challenge re-infection. The level of complement-dependent treponemicidal activity in sera obtained before infection (basal) and before intradermal challenge was determined by the "washed-killing" assay and compared with that detected using conventional in vitro immobilization. RESULTS: Using the "washed-killing" assay, a close quantitative correlation as measured by a treponemal immobilizing endpoint titer was demonstrable between prechallenge treponemicidal antibody and the status of immunity to re-infection. Sera from rabbits completely susceptible to symptomatic and disseminated asymptomatic re-infection lacked treponemicidal antibody. Sera from challenged rabbits with a relatively low degree of immunity to symptomatic disease showed endpoints of < or = 4. Rabbits with a relatively high degree of immunity to symptomatic reinfection and resistant to disseminated disease had endpoints that ranged from 6 to 96. Rabbits completely resistant to challenge exhibited endpoints ranging from 96 to 128. CONCLUSION: Treponemicidal antibody measured by the "washed-killing" assay correlated closely with the status of immunity in experimental rabbit syphilis. Thus, antibody measured by this assay may be directed against key protective Treponema pallidum surface immunogens.

Animals↗

Vaginal washing and increased risk of HIV-1 acquisition among African women: a 10-year prospective study.

BACKGROUND: No prospective study has examined the risk of HIV-1 acquisition associated with vaginal washing, although intravaginal practices have been identified as potentially important contributors to HIV-1 susceptibility. OBJECTIVE: To evaluate the contribution of vaginal washing to incident HIV-1 infection. DESIGN: Prospective cohort study. METHODS: Data were derived from a 10-year study of risk factors for HIV-1 acquisition among 1270 Kenyan female sex workers. Intravaginal practices were ascertained at study enrollment. At monthly follow-up visits, women completed a standardized interview and specimens were collected for diagnosis of HIV-1 and genital tract infections. RESULTS: : Compared with women who did not perform vaginal washing, there was an increased risk for acquiring HIV-1 among women who used water [adjusted hazard ratio (HR), 2.64; 95% confidence interval (CI), 1.00-6.97] or soap (adjusted HR 3.84; 95% CI, 1.51-9.77) to clean inside the vagina, after adjustment for demographic factors, sexual behavior, and sexually transmitted infections. Furthermore, women who performed vaginal washing with soap or other substances were at higher risk for HIV-1 compared with those who used water alone (adjusted HR, 1.47; 95% CI, 1.02-2.13). CONCLUSIONS: In populations where vaginal washing is common, this practice may be an important factor promoting the spread of HIV-1. Intervention strategies aimed at modifying intravaginal practices should be evaluated as a possible female-controlled HIV-1 prevention strategy.

Adult↗

Metabolism of red beet slices I. Effects of washing.

The changes in relative participation of pathways of glucose catabolism in red beet slices during washing have been examined using specifically (14)C labeled glucoses. Washing of these slices brings about an increase in participation of the pentose phosphate pathway. The composition of the washing medium influences slightly the extent of change in pathway participation. The activity level of certain enzymes participating in the initial stages of glucose catabolism has been measured in fresh and washed beet slices. Fresh slices which barely metabolized gluconate were found to have very little 6-phosphogluconate dehydrogenase activity. Washing brings about a dramatic increase in 6-phosphogluconate dehydrogenase activity and this increase was accompanied by a marked increase in the ability of the slices to metabolize gluconate. In red beet slices the TPNH generated via pentose phosphate pathway appears to be utilized for biosynthetic reductions rather than as respiratory substrate.

Journal Article↗

Study of the efficacy of antiseptic handrub lotions with hand washing machines.

The effectiveness of quick handwashing in regards to four alcohol-based handrub lotions, including two products already on the market and our original lotions 1, 2, with two different kinds of hand washing machines was studied in vivo. We also tried to evaluate the efficacy of the four lotions in vitro. The in vivo testing of each lotion, including ethanol, was evaluated using two hand washing methods, first with and then without a previous soap wash. Computerized image analysis was used to calculate the bacterial count on the hand surface. The results showed that a 3 second application of the original lotion 2 (0.5% chlorhexidine in a 77% ethyl alcohol solution) preceded by a soap wash eradicated 89.3% of the bacteria on the hands; this was the highest reduction rate of all four detergents. AROKULIN-E (67.9% ethyl alcohol solution) without a prior handwashing produced the lowest reduction, 49.1% against hand surface bacteria. These results were compatible with those in vitro. It seems that an alcohol-based solution containing an effective antimicrobial detergent preceded by a soap wash is necessary to acceptably reduce hand surface bacteria.

Bacteria↗

Evidence that endotoxins enhance the factor X activator activity of washed human platelets.

The in vitro effect of several bacterial endotoxins on human platelets was determined. Nine different endotoxins failed to induce aggregation in platelet-rich plasma (PRP) or of platelets washed by two different methods; four of them which we studied further failed to induce [14C]serotonin release in PRP. In contrast, using recently described test systems for platelet coagulant activity, all the endotoxins shortened the latent period occurring before aggregation of a mixture of washed platelets, normal serum, and CaCl2, and the clotting time of this mixture upon addition of fibrinogen. Washed platelets obtained from PRP preincubated with endotoxin had a higher platelet coagulant activity than platelets obtained from PRP preincubated with buffer. Washed platelets contribute to thrombin generation by providing factor V, a factor X activator and possibly phospholipid. Since the endotoxins did not influence the factor V activity of platelets or the platelet factor 3 activity, either in PRP or using platelets washed by albumin density gradient centrifugation, it is suggested that they enhance the factor-X activator activity of human platelets.

Blood Platelets↗