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PubMed1972

Dietary protein intake and skeletal-muscle protein metabolism in rats. Studies with salt-washed ribosomes and transfer factors.

1. Aspects of skeletal muscle protein synthesis in vitro were studied in young rats given a low-protein diet for up to 10 days and during re-feeding with an adequate diet. 2. Partially purified muscle transfer factors (transferases I and II), crude and purified (NH(4)Cl-washed) ribosomes and a pH5 enzyme fraction were prepared for this purpose. 3. A marked decrease in the capacity of crude ribosomes to carry out cell-free polypeptide synthesis occurred within 4 days of feeding the low-protein diet. 4. The capacity of salt-washed ribosomes to promote amino acid polymerization, in the presence of added transfer factors and aminoacyl-tRNA, was only slightly decreased by the dietary treatment. 5. However, the capacity of salt-washed ribosomes to bind (14)C-labelled aminoacyl-tRNA was decreased by feeding the low-protein diet. 6. The capacity of the pH5 enzyme fraction to promote amino acid incorporation in a complete cell-free system was decreased within 2 days of feeding the low-protein diet. There is no evidence that the change is associated with aminoacyl-tRNA synthetase or binding enzyme activities of the pH5 fractions. 7. These changes are discussed in relation to the diminished rate of protein synthesis in the intact muscle cell when rats are given a low-protein diet.

Acyltransferases↗

PubMed1999

Methanogenic archaea and CO2-dependent methanogenesis on washed rice roots.

Washed excised roots of rice (Oryza sativa) immediately started to produce CH4 when they were incubated in phosphate buffer under anoxic conditions (N2 atmosphere), with initial rates varying between 2 and 70nmolh(-1)g(-1) dry weight of root material (mean +/- SE: 20.3 +/- 5.9 nmol h(-1) g(-1) dry weight; n = 18). Production of CH4 continued for at least 500 h, with rates usually decreasing slowly. CH4 production was not significantly affected by methyl fluoride, an inhibitor of acetoclastic methanogenesis. Less than 0.5% of added [2-14C]-acetate was converted to 14CH4, and conversion of 14CO2 to 14CH4 indicated that CH4 was almost exclusively produced from CO2. Occasionally, however, especially when the roots were incubated without additional buffer, CH4 production started to accelerate after about 200h reaching rates of > 100 nmol h(-1) g(-1) dry weight. Methyl fluoride inhibited methanogenesis by more than 20% only in these cases, and the conversion of 14CO2 to 14CH4 decreased. These results indicate that CO2-dependent rather than acetoclastic methanogenesis was primarily responsible for CH4 production in anoxically incubated rice roots. Determination of most probable numbers of methanogens on washed roots showed highest numbers (10(6)g(-1) dry roots) on H2 and ethanol, i.e. substrates that support CH4 production from CO2. Numbers on acetate (10(5) g(-1) dry roots) and methanol (10(4)g(-1) dry roots) were lower. Methanogenic consortia enriched on H2 and ethanol were characterized phylogenetically by comparative sequence analysis of archaeal small-subunit (SSU) ribosomal RNA-encoding genes (rDNA). These sequences showed a high similarity to SSU rDNA clones that had been obtained previously by direct extraction of total DNA from washed rice roots. The SSU rDNA sequences recovered from the H2/CO2-using consortium either belonged to a novel lineage of methanogens that grouped within the phylogenetic radiation of the Methanosarcinales and Methanomicrobiales or were affiliated with Methanobacterium bryantii. SSU rDNA sequences retrieved from the ethanol-using consortium either grouped within the genus Methanosarcina or belonged to another novel lineage within the phylogenetic radiation of the Methanosarcinales and Methanomicrobiales. Cultured organisms belonging to either of the two novel lineages have not been reported yet.

Acetates↗

PubMed2001

Mitomycin-supplemented washed blood agar for the isolation of Shiga toxin-producing Escherichia coli other than O157:H7.

AIMS: Isolation and recognition of the prominent Shiga toxin (Stx)-producing strains of Escherichia coli (STEC) serovar O157:H7 can be confirmed easily by their late fermentation of sorbitol and lack of beta-glucuronidase activity, but there has been no culture method of choice for detecting non-O157 STEC strains because of their biochemical diversity. Apart from Stx, many STEC strains produce enterohaemolysin (Ehly) regardless of their serovars. METHODS AND RESULTS: Although washed blood agar media, with or without the addition of antibiotics (vancomycin, cefixime, and cefsulodin) (WBA and WBVCCA), have been used to detect Ehly, a proportion of STEC strains consistently failed to produce haemolysin on these media. Washed blood agar medium was therefore studied further in order to increase the yield of strains producing Ehly. CONCLUSION: It was found that the addition of 0.5 microg ml(-1) of mitomycin C to the agar medium (WBMA) markedly increased the number of such strains. Thus, of 185 STEC strains comprising 95 O157 and 90 non-O157 STEC consisting of 34 serovars. Ninety-seven per cent of these strains produced haemolysis on WBMA, compared with only 76% and 83%, respectively, on WBA and WBVCCA. SIGNIFICANCE AND IMPACT OF THE STUDY: The appearance of the Ehly zone of haemolysis that was easily distinguishable from that of alpha-haemolysin was enhanced by the incorporation of mitimycin C into washed-blood medium.

Blood↗

PubMedDate not supplied

Viable lymphocytes in frozen washed blood.

Samples of glycerolized, frozen, packed cells, were washed by each of three systems and then were cultrued for viable lymphocytes using the short-term culture method. Of the 39 samples, 17 (43.6%) showed definite evidence of growth of lymphocytes. Of the 17, 11(64.7%) were washed by Elutramatic, two (11.8%) by Haemonetics, and four (23.5%) by IBM. The 4 C prefreeze (two to five days) as well as the -80 C (21 To 40 days) storage times were the same for both positive and negative specimens. In all samples, clumps of distorted granulocytes with pyknotic nuclei were seen in addition to a number of well-preserved mononucleated white cells. In the 17 specimens which showed growth, incorporation of 3H-thymidine was seen only in the PHA-induced blast cells; typical mitotic figures were seen in some cultures. These observations demonstrate that lymphocytes remain viable in frozen blood stored at -80 C up to 40 days. The significant difference in removal of viable lymphocytes noted in specimens washed by different instruments requires further evaluation.

Blood Preservation↗

PubMedDate not supplied

Exchange transfusion in Reye's syndrome with saline-washed red blood cells.

Reye's syndrome (RS) consists of encephalopathy, fatty degeneration of viscera, and elevation of ammonia, certain amino acids, and liver enzymes in the blood. It is most characteristically a disease of children and has been considered to have a poor prognosis despite the use of various treatment regimens. Exchange transfusion (ET) with fresh blood (less than 24 hours old) for the removal of toxic metabolic byproducts in TS is a relatively recent development which appears to have improved the survival rates. However, because RS may occur in epidemic proportions at any time the demand for fresh blood can place an excessive stress on blood resources. We have, therefore, utilized saline-washed (to remove potentially toxic metabolites) red blood cells (RBC's) less than six days old, and fresh frozen citrate-phosphate-dextrose (CPD) plasma in treating this disorder. Nine patients in the Milwaukee community with severe encephalopathy secondary to RS were treated with ET. These patients collectively required 151 units of washed RBC's for 21 ET. Eight of nine patients survived without sequelae. Age of blood used did not correlate with the number of exchanges required or the eventual outcome. A continual computerized EEG in five patients provided objective evidence of the effectiveness of ET. We conclude that ET with saline-washed RBC's reconstituted with fresh frozen plasma (FFP) is a fast, safe, and effective means of treating patients with RS.

Brain Diseases↗

PubMedDate not supplied

The preparation of leukocyte-poor red cells from liquid stored blood: an evaluation of the Haemonetics 102 cell washing system.

An evaluation of the Haemonetics 102, an automated cell washing system, is presented. Although the system removes 68.4 +/- 14 per cent of the white blood cells from liquid stored blood, the percentage wash out is an unreliable mode of evaluation. The residual number of white blood cells per unit is 6.5 +/- 2.8 x 10(8), which is independent of the unit's size, age, and initial leukocyte count. The residual number of white blood cells is also not influenced by any of the modifications used in the study. Red blood cell loss averaged 16.3 per cent. A direct correlation exists between the initial red blood cell mass and the amount of cell lost. It is recommended that packed cell units which have a volume of less than 300 ml with hematocrits of 75 per cent or less be used to minimize excessive red blood cell loss. Recommendation as to the selection of units by an ideal age or the preparation of units prior to washing are not supported by these data.

Blood Preservation↗

PubMedDate not supplied

Platelet washing with a blood cell processor.

Washing of platelet components for transfusion may be indicated in selected patients. This study was designed to determine a protocol using an automated cell processor to remove plasma proteins from pooled platelet concentrates, stored at 20 to 24 degrees C with constant rotation. A 1500-ml, 0.9% saline-wash procedure is described. Eight percent of the platelets were lost with the procedure (range, 0-19%), while a mean of 99.6 percent of the plasma protein was removed (range, 99.2-99.8%). The clinical effectiveness of maternal platelets washed by this method was demonstrated in a case of alloimmune neonatal thrombocytopenia.

Adult↗

PubMed1991

The neonatal immune response to washed and irradiated red cells: lack of evidence of lymphocyte activation.

A total of 21 neonatal infants (11 males and 10 females) were evaluated for lymphocyte subpopulation changes and cellular activation following the receipt of washed and gamma-irradiated red cells. The mean donor exposure was 2 +/- 1.5 donors, and the mean white cell concentration of each 10-mL-per-kg transfusion was 2 x 10(7) cells. A total of 2800 rad (28 Gy) was delivered to each red cell unit prior to washing. The lymphocyte subsets CD3+, CD4+, CD8+, CD19+, CD3+/CD25+, CD3+/HLA-DR, CD4+/CD45RA, CD4+/CDw29, and CD4+/ICHL-1 were analyzed, as were plasma gamma-interferon and neopterin levels, before and after blood transfusion. Posttransfusion samples were obtained from 3 to 12 days after the last blood transfusion. There were no posttransfusion changes in the percentage of lymphocyte subsets analyzed. Furthermore, overlay-histogram analysis of pretransfusion and posttransfusion CD45RA-, CDw29-, and UCHL-1-positive helper T cells failed to reveal a shift in mean channel fluorescence intensity, which is indicative of a lack of cellular activation. Gamma-interferon levels remained unchanged after transfusion (range, 90 +/- 5.9 to 95.5 +/- 5.3 pg/mL), as did neopterin levels (range, 3.7 +/- 1.5 to 4.6 +/- 1.5 ng/mL). This study could not find any evidence, by either cellular or humoral markers, of the activation of neonatal lymphocytes by transfusion. It is hypothesized that this failure to observe lymphocyte activation is due to the low number of donor white cells present in washed, irradiated red cells and/or to a lack of recipient recognition of transfused, allogeneic, donor white cells.

Blood Transfusion↗

PubMed1994

Patient and staff exposure to glutaraldehyde from KeyMed Auto-Disinfector endoscope washing machine.

Activated glutaraldehyde (2%) is the recommended agent for disinfection of endoscopic equipment. Exposure to the disinfectant is associated with side-effects and guidelines have been set to avoid these complications. Endoscope washing machines are used to provide automated high level disinfection of endoscopes as well as reduce exposure to the irritant aldehyde. We report a design fault in an endoscope washing machine which results in patients and staff being exposed to activated glutaraldehyde. The Auto-Disinfector (KeyMed) comprises a washing chamber supplied by three separate reservoirs (detergent, disinfectant, and rinse water) via a common channel. After the first cycle endoscopes are processed using previously used detergent and rinse water. Rinse water glutaraldehyde concentrations were measured in four machines during routine endoscopy lists on at least two occasions and showed a progressive rise in glutaraldehyde concentration up to 0.1% after two to six cycles. This results in staff being exposed to glutaraldehyde present on the processed instrument and the disinfectant being pumped from the internal channels of the endoscope into direct contact with the gastrointestinal mucosa during endoscopy at concentrations of 200-1000 ppm. The present atmospheric limit for glutaraldehyde is 0.2 ppm. Skin and mucosal irritation occur at concentrations of 0.3 ppm and severe synovitis in experimental animals at 100 pm. Tongue swelling and bloody diarrhoea with characteristic mucosal histological changes have been reported in patients exposed to activated glutaraldehyde on inadequately rinsed equipment. To reduce this problem the rinse water should be changed after every cycle and the endoscope should be dried thoroughly before use.

Disinfection↗

PubMed1995

Recovery of intraoperatively shed blood in aortoiliac surgery: comparison of cell washing with simple filtration.

To regain blood shed intraoperatively, two different systems are clinically established: washing and centrifuging red blood cells to produce autologous erythrocyte concentrates and devices for immediate reinfusion of whole blood after mere filtration. In a prospective-randomised study to compare both methods regarding their efficiency, adverse effects, and economy, 20 patients of our department undergoing elective aortoiliac surgery received intraoperative autotransfusion by means of either cell-washing (CS) or salvage of whole blood (WB). Patients were preoperatively randomized into one of the two groups and were evaluated with respect to standard metabolic and haematological laboratory parameters preoperatively, during surgery, after transfer into the recovery room, 24 h after surgery, after transfer into the recovery room, 24 h after surgery, and at discharge. Both patient groups were well comparable in demographics, preoperative laboratory data, and indication for operation. Handling was easier, the set-up time was shorter with the whole blood filtration device (10.2 +/- 2.3 versus 21 +/- 1.9 min, p = 0.0023), and no additional personnel was needed to run the system. The whole blood device also allowed a greater percentage of aspirated blood to be returned intraoperatively compared to cell washing (73.5% +/- 7.2 versus 51.1% +/- 6.5, p = 0.03). Thrombocytopenia occurred in 7 (CS) and 3 (WB) patients intraoperatively with a significant difference in platelet count between the two groups (118 +/- 17 [CS] versus 170 +/- 12 [WB]*10(9)/L, p = 0.025). Expected changes in the perioperative course of the clotting parameters such as highly increased PTT levels and moderately prolonged prothrombin times occurred in all cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

PubMed1979

Hepatitis after the transfusion of frozen red cells and washed red cells.

The large-scale use of frozen red cells and washed red cells in this transfusion service has not been effective in reducing post-transfusion hepatitis. Seventy-eight per cent of 31,125 transfusions, excluding platelets and plasma, were given in the form of frozen or washed red cells. These transfusions were associated with 56 cases of hepatitis. In 37 cases, the recipients had received only frozen or washed red cells (or both). Post-transfusion hepatitis was of the non-A, non-B type in 95% of cases. On average, commercial blood accounted for 30 per cent of the donor base during the first four years and was involved in 44 of the 56 cases. Elimination of commercial blood from the donor base resulted in a decrease in the incidence of hepatitis from 2.1 to 0.3 per 1000 transfusions.

Blood Banks↗

PubMed2004

Residues of azoxystrobin, fenhexamid and pyrimethanil in strawberry following field treatments and the effect of domestic washing.

Residues of the pesticides azoxystrobin, fenhexamid and pyrimethanil were determined in strawberry after field treatment. The effect of 'home' washing with tap water and a commercially available vegetable detergent on residue levels was also studied. After treatment, azoxystrobin and pyrimethanil residues on strawberry were on average 0.55 and 2.98 mg kg(-1), respectively, values below the maximum residue level (MRL) fixed by the European Union (2.0 and 5 mg kg(-1), respectively), while fenhexamid residues were on average 2.99 mg kg(-1), which is very close to the MRL of 3.0 mg kg(-1), but some samples were over the MRL. Thereafter, all residues declined, with a half-life of about 8 days (azoxystrobin and fenhexamid) and 4.8 days (pyrimethanil). Washing the fruit with tap water reduced the residues of azoxystrobin and fenhexamid but did not affect pyrimethanil residues. Finally, when fruits were washed with a commercial detergent, greater amounts were removed (about 45% of azoxystrobin and pyrimethanil and 60% of fenhexamid).

Acrylates↗

PubMed2001

Biological treatment of fishery washing water using Bacillus sphaericus coupled with production of spores that are toxic to mosquito larvae.

Research was undertaken to investigate the treatment of fishery washing water using Bacillus sphaericus, and to recover the spores for subsequent use as bioinsecticide to control the population of mosquitoes. This treatment method could reduce pollution due to organic matter by decreasing the value of Chemical Oxygen Demand (COD) and Biological Oxygen Demand (BOD) by about 85% and 92%, respectively. The maximum concentration of spores (83.3 x 10(7) spores ml(-1)) using normal concentration of filtered fishery washing water was only about 27% lower than that obtained in fermentation using 0.25% (w/v) yeast extract. The larvicidal activity of the spores produced in fermentation using fishery washing water to Culex quinquefaciatus, as measured by LD50 after 48 h, was almost the same as the larvicidal activity of spores obtained from fermentation using yeast extract.

Animals↗

PubMed2001

Development of sampling and analytical methods for concerted determination of commonly used chloroacetanilide, chlorotriazine, and 2,4-D herbicides in hand-wash, dermal-patch, and air samples.

Sampling and analytical methods were developed for commonly used chloroacetanilide, chlorotriazine, and 2,4-D herbicides in hand washes, on dermal patches, and in air. Eight herbicides selected for study were alachlor, atrazine, cyanazine, 2,4-dichlorophenoxyacetic acid (2,4-D), metolachlor, simazine, and two esters of 2,4-D, the 2-butoxyethyl ester (2,4-D, BE) and the 2-ethylhexyl ester (2,4-D, EH). The hand-wash method consisted of shaking the worker's hand in 150 mL of isopropanol in a polyethylene bag for 30 seconds. The dermal-patch method entailed attaching a 10-cm x 10-cm x 0.6-cm polyurethane foam (PUF) patch to the worker for exposure; recovery of the herbicides was achieved by extraction with 40 mL of isopropanol. The air method involved sampling with an OVS-2 tube (which contained an 11-mm quartz fiber filter and two beds of XAD-2 resin) and recovery with 2 mL of 10:90 methanol:methyl t-butyl ether. Analysis of each of the three sample types was performed by gas chromatography with an electron-capture detector. Diazomethane in solution was employed to convert 2,4-D as the free acid to the methyl ester in each of the three methods for ease of gas chromatography. Silicic acid was added to sample solutions to quench excess diazomethane. Limits of detection for all eight herbicides were matrix-dependent and, generally, less than 1 microgram per sample for each matrix. Sampling and analytical methods met NIOSH evaluation criteria for all herbicides in hand-wash samples, for seven herbicides in air samples (all herbicides except cyanazine), and for six herbicides in dermal-patch samples (all herbicides except cyanazine and 2,4-D). Speciation of 2,4-D esters and simultaneous determination of 2,4-D acid were possible without losses of the esters or of other herbicides (acetanilides and triazines) being determined.

2,4-Dichlorophenoxyacetic Acid↗

PubMed2003

Isolation and in vitro cultivation of human urothelial cells from bladder washings of adult patients and children.

To acquire urothelial cells for in vitro engineering of urothelium, biopsy specimens were taken from the urological tract. In clinical practice the number of cells harvested by biopsy are limited and the procedure requires general anaesthesia in children. The purpose of this study was to find out if bladder washings from adult patients as well as children contained enough proliferative and colony-forming uroepithelial cells to regenerate urethral mucosa in vitro, and if the cells could be stored by freezing. Bladder washings from nine children and eight adult patients were collected from patients who were having procedures that required an indwelling catheter. All cultures grew colonies of cells with a morphological appearance typical for epithelial cell growth. The cultures could be expanded to confluent, stratified sheets, and cells that stained for pancytokeratin, indicating an epithelial origin. Cells stored in -150 degrees C could be cultured and expanded in vitro. No differences were seen between cells from adults and children. Bladder washing is a non-invasive way to obtain many autologous urothelial cells. The method is reproducible and well tolerated by children. The possibility of culturing cells obtained in this way into stratified grafts provides a unique way of reconstructing the urogenital tract by "tissue engineering".

Adult↗

PubMed2004

Washing of soil contaminated with PAHs and heavy petroleum fractions using two nonionic [correction] and one ionic surfactant: effect of salt addition.

The aim of this work was to select the surfactant for washing an aged soil contaminated with PAHs and heavy hydrocarbons fraction, between two anionic and one ionic surfactant. The effect of the addition of salt when washing with the ionic surfactant was evaluated. Results of this work indicated the following. Soil contaminants concentration as g TPH kg soil(-1) was 17,200 mg kg(-1). All washing performances were dependent on the type of surfactant and applied doses. In general, best removal efficiencies were obtained with TW80 (59.57%), followed very near by E600 (57.67%), and at the en by SDS (39.0%), in comparison to the blank assessment (only water), where removal efficiency was about 12%. These removal efficiencies were correlated to the MW, CMC, and HLB values of every surfactant. SDS efficiencies were quite enhanced by the addition of 1% (60.36% removal efficiency) and 2% (70.28% removal efficiency) of NaCl. Its seems that there exist two reasons for this phenomena i.e., the diminution in the real value of SDS CMC (as reported by many authors) and, the capabilities of NaCl solutions to dissolve higher amounts of hydrocarbons than only tap water.

Environmental Pollution↗

PubMed2001

The use of oral washes to diagnose Pneumocystis carinii pneumonia: a blinded prospective study using a polymerase chain reaction-based detection system.

Pneumocystis carinii pneumonia (PCP) can be diagnosed by direct microscopic examination of induced sputum or by bronchoalveolar lavage (BAL). However, many institutions have little diagnostic success with induced sputum, and BAL is invasive and expensive. This prospective, blinded study assessed oral washes as a more convenient specimen than either sputum or BAL fluid and used a dissociation-enhanced lanthanide fluoroimmunoassay time-resolved fluorescent hybridization polymerase chain reaction (PCR) detection system that is feasible for clinical laboratories. The study assessed 175 oral washes, each paired with either an induced sputum that was positive for Pneumocystis or a BAL sample. The PCR test based on the Pneumocystis major surface glycoprotein primers had a sensitivity of 91% and a specificity of 94%, compared with a test based on mitochondrial large subunit rRNA primers, which had a sensitivity of 75% and a specificity of 96%. These results suggest that oral washes can provide a useful sample for diagnosis of PCP when a sensitive PCR detection system is used.

Bronchoalveolar Lavage↗

PubMed2000

Skin irritation and dryness associated with two hand-hygiene regimens: soap-and-water hand washing versus hand antisepsis with an alcoholic hand gel.

OBJECTIVE: To compare the frequency of skin irritation and dryness associated with using an alcoholic-hand-gel regimen for hand antisepsis versus using soap and water for hand washing. DESIGN: Prospective randomized trial with crossover design. Irritation and dryness of nurses' hands were evaluated by self-assessment and by visual assessment by a study nurse. Epidermal water content of the dorsal surface of nurses' hands was estimated by measuring electrical capacitance of the skin. SETTING: Miriam Hospital, a 200-bed university-affiliated teaching hospital. PARTICIPANTS: Thirty-two nurses working on three hospital wards participated in the trial, which lasted 6 weeks. RESULTS: Self-assessment scores of skin irritation and dryness decreased slightly during the 2 weeks when nurses used the alcoholic-hand-gel regimen (mean baseline score, 2.72; mean final score, 2.0; P=.08) but increased substantially during the 2 weeks when nurses used soap and water (mean baseline score, 2.0; mean final score, 4.8; P<.0001). Visual assessment scores by the study nurse of skin irritation and dryness did not change significantly when the alcoholic-hand-gel regimen was used (mean baseline and final scores were both 0.55), but scores increased substantially when nurses used soap and water (baseline score, 0.59; mean final score, 1.21; P=.05). Epidermal water content of the dorsal surface of nurses' hands changed little when the alcoholic-hand-gel regimen was used (mean+/-standard deviation baseline electrical capacitance reading, 24.8+/-6.8; mean final reading, 25.7+/-7.3), but decreased significantly (skin became dryer) with soap-and-water hand washing (mean baseline, 25.9+/-7.5; mean final reading, 20.5+/-5.4; P=.0003). CONCLUSIONS: Hand antisepsis with an alcoholic-hand-gel regimen was well tolerated and did not result in skin irritation and dryness of nurses' hands. In contrast, skin irritation and dryness increased significantly when nurses washed their hands with the unmedicated soap product available in the hospital. Newer alcoholic hand gels that are tolerated better than soap may be more acceptable to staff and may lead to improved hand-hygiene practices.

Antisepsis↗