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Elastase levels in salvaged blood and the effect of cell washing.

Blood was salvaged from the operating field of 16 orthopaedic and vascular operations and processed by the Shiley Dideco cell saver for retransfusion. Plasma elastase levels in the salvaged blood were used as a marker of white cell lysosomal granule release. The plasma level of this enzyme was measured by the technique described by Dreher et al. using the immuno-activation (IMAC) system (Merck-Darmstadt, Federal Republic of Germany). The levels of the enzyme rose dramatically in the salvaged blood and were reduced to preoperative levels after washing in the cell saver. Washing of the salvaged blood before retransfusion is effective in reducing the plasma levels of white cell lysosomal contents which have the potential for producing harmful systemic effects.

Blood Transfusion, Autologous↗

Serum albumin washing specifically enhances arachidonate incorporation into synaptosomal phosphatidylinositols.

Arachidonate incorporation into synaptosomal phospholipids was shown to be affected by factors including the procedure for preparation of the membrane fractions and preincubation of synaptosomes prior to assay of incorporation of arachidonate into both phosphatidylcholine (PC) and phosphatidylinositol (PI). However, the inhibition toward incorporation into PIs, but not PCs, was fully reversed when the membranes were washed with bovine serum albumin. A twofold increase in arachidonate incorporation into PIs was also observed when freshly prepared synaptosomes were washed with serum albumin immediately before assay of incorporation activity. The inhibitory action is thought to be due to an increase in polyunsaturated fatty acids and/or their oxidation products which may then elicit a special effect on the acyltransferase responsible for transferring arachidonate into phosphatidylinositols. The differences in fatty acid uptake and response to serum albumin also suggest the presence of different acyltransferase for acyl transfer to PIs and PCs.

Animals↗

Increase of plasma renin activity at renal blood flow estimations with the xenon133 wash-out technique in patients with renal artery stenosis.

Eleven patients with hypertension and renal artery stenosis of fibromuscular type were investigated with renal blood flow estimations with Xenon133 wash-out technique before and after alpha-receptor blockade with infusion of phenoxybenzamine in the stenosed renal artery. During the whole investigation frequent blood samples for renal vein renin estimations were drawn from both kidneys. Immediately after the Xenon injection renal vein renin elevations on the investigated side were observed in 9/11 patients. The renin peak was reached in 10 min and the mean increase for all patients was 152%. The mean duration of the peak was 19.25 min. A neurogenic renin release mechanism triggered by renal chemoreceptors and mediated by beta-adrenergic fibres was suggested. This finding with renin release stimulation at Xenon133 infusion in the renal artery is of great importance in judging results of investigations concerning renal blood flow and renin when the Xenon wash-out technique is used. Blood samples for renin estimations should not be taken within 25 min after intrarenal Xenon133 infusions.

Adult↗

Does abnormal single-breath nitrogen wash-out predict an accelerated decline in FEV1 in lung-healthy subjects?

Single-breath nitrogen wash-out and forced expirograms measured on two occasions, 8 years apart, were analysed for 24 subjects with varying degrees of peripheral airway involvement. Correlation analyses were carried out between closing volume in percentage of vital capacity (CV%), the slope of the alveolar plateau (phase III) and forced expiratory volume in 1 s (FEV1) for both occasions. The correlations between CV% and phase III measured on the first occasion and the long-term change in FEV1 were also studied. It is concluded that large deviations are required for CV% as well as phase III before any pathological decreases in the large airways will occur. The prognostic value of an abnormal single-breath nitrogen wash-out seems to be limited, at least in lung-healthy subjects.

Adult↗

Peritoneal washing cytology: a retrospective analysis of 175 gynaecological patients.

Accurate staging is the basis for optimal therapy in the management of the gynaecological cancer patient and, in recent years, peritoneal washings are one of the techniques used to determine this, both at the initial and subsequent laparotomies. Two hundred and four peritoneal washes were obtained from 175 patients during a 5-year period. Fifteen cases of histologically proven intraperitoneal tumour were confirmed in 11 cases by periotoneal cytology at initial operation, a sensitivity of 73%. At second-look laparotomy, 3 of 9 histologically proven malignancies were confirmed by cytology, a sensitivity of 33%. One false positive diagnosis was made in the series. Procedures to avoid both false-positive and false-negative diagnoses by the clinician and the laboratory are discussed, and a review of the literatue is provided.

Cytodiagnosis↗

Microaggregates in frozen and saline washed red blood cells.

The microaggregates which accumulate in stored blood have been implicated in the development of posttraumatic pulmonary insufficiency. These particles are known to be composed of degenerated leukocytes and platelets. Because frozen and saline washed red blood cells contain small numbers of leukocytes and platelets, they were studied as a possible source of microaggregate-free red blood cells. Using a Model T Coulter Counter to quantitate all particles 13-80 microns in size, it was shown that freezing and deglycerolization, or simple saline washing (manual or automated), could reduce the number of microaggregates in stored blood by 80 to 90 per cent. These findings add to a growing list of potential advantages in the routine use of frozen red cells for patients requiring transfusion.

Erythrocyte Aggregation↗

The roles of ADP and TXA in botrocetin/VWF-induced aggregation of washed platelets.

BACKGROUND: Binding of von Willebrand factor (VWF) to the platelet membrane glycoprotein (GP) Ib-IX-V complex initiates a cascade of events leading to alphaIIbbeta3 activation and platelet aggregation. The roles of ADP and thromboxane A2 (TXA2) in agglutination-induced GPIbalpha-mediated platelet activation have not been fully described. METHODS: Botrocetin and human VWF were used to stimulate washed mouse platelets. Platelets deficient in TXA2 receptors, Galphaq, or alphaIIbbeta3, and inhibitors and chelating agents were used to investigate the roles of TXA2, ADP, alphaIIbbeta3 and Ca2+ in botrocetin/VWF-induced signaling. RESULTS: Our data demonstrate that botrocetin/VWF/GPIbalpha-mediated agglutination results in calcium-independent protein kinase C (PKC) and phospholipase A2 (PLA2) activities required for GPIbalpha-elicited TXA2 production that in turn causes dense granule secretion. Aggregation of washed platelets requires TXA2-induced alphaIIbbeta3 activation and ADP signaling. TXA2 or ADP can activate alphaIIbbeta3, but both are required for alpha-granule secretion and aggregation. Botrocetin/VWF-induced dense granule secretion is Galphaq-dependent. alpha-Granule secretion requires initial ADP signaling through P2Y1 and subsequent signaling through P2Y12. Signaling initiated by agglutination is propagated and amplified in an alphaIIbbeta3-dependent manner. CONCLUSIONS: In contrast to adhesion or shear stress-induced GPIb-elicited signaling, agglutination-elicited GPIb signaling that activates alphaIIbbeta3 requires TXA2. Agglutination-elicited TXA2 production is independent of Ca2+ influx and mobilization of internal Ca2+ stores. Therefore, our results demonstrate that agglutination-elicited GPIb signaling causes alphaIIbbeta3 activation by a mechanism that is distinct from those used by adhesion, or shear stress-induced GPIb signaling.

Adenosine Diphosphate↗

Anaerobic conversion of carbon dioxide to methane, acetate and propionate on washed rice roots.

Washed excised roots of rice (Oryza sativa) produced H(2), CH(4), acetate, propionate and butyrate when incubated under anoxic conditions. Acetate production was most pronounced with a maximum rate (mean+/-standard error; four different root preparations) of 3.4+/-0.6 µmol h(-1) g-dry weight(-1) roots, compared to 0.45+/-0.13, 0.06+/-0.03, and 0.04+/-0.01 µmol h(-1) g-dw(-1) for propionate, butyrate and CH(4)1 kPa after one day of incubation. Then it decreased and reached more or less constant concentrations of about 50-80 Pa after about 7-8 days. Hydrogen partial pressures were always high enough to allow exergonic methanogenesis (DeltaG=-67 to -98 kJ mol(-1) CH(4)) and exergonic homoacetogenesis (DeltaG=-18 to -48 kJ mol(-1) acetate) from H(2) plus CO(2). Radioactive bicarbonate/CO(2) was incorporated into CH(4), acetate and propionate. The specific radioactivities of the products indicated that CH(4) was exclusively produced from H(2)/CO(2) confirming a previous study. The contribution of CO(2) to the production of acetate and propionate was 32-39% and 42-61%, respectively, assuming that each carbon atom was equally labeled. Propionate also became radioactively labeled, when the roots were incubated with either [1-(14)C]acetate or [2-(14)C]acetate accounting for 60-76% of total propionate production. Reductive formation of propionate was thermodynamically favorable both from H(2) plus acetate plus CO(2) (DeltaG=-15 to -38 kJ mol(-1) propionate) and from H(2) plus CO(2) (DeltaG=-34 to -85 kJ mol(-1) propionate). A substantial fraction of propionate was apparently reductively formed from acetate and/or CO(2). In conclusion, our results demonstrate an intensive anaerobic dark metabolism of CO(2) on washed rice roots with reduction of CO(2) contributing significantly to the production of acetate, propionate and CH(4). The CO(2) reduction seemed to be driven by decay and fermentation of root material.

Journal Article↗

Tracheobronchial washings from seven vertebrate species as growth media for the four species of Bordetella.

The four species of Bordetella differed in their ability to grow at 37 degrees C in membrane-filtered tracheobronchial washings (TBW) from seven vertebrate species, including their natural hosts. From washed inocula of approximately 2 x 10(3) colony-forming units per ml (cfu ml-1), Bordetella bronchiseptica and B. avium grew much better than the other two bordetellae and yielded stationary-phase cultures containing 10(8)-10(9) cfu ml-1 in most of the TBW samples. These counts were only moderately lower than those attained in CL medium which contains about a 450-times higher concentration of amino acids. B. bronchiseptica and B. avium also grew to a limited extent in phosphate-buffered saline without nutrient supplements. B. parapertussis grew in TBW from man, sheep, rabbit, mouse and chicken, but not in TBW from a dog and a horse or in PBS. B. pertussis grew well in CL medium, but not in PBS or in any of 13 samples of TBW from the seven vertebrate species, which included three samples of lung lavage fluid from human patients. Analysis of the TBW samples for known Bordetella nutrients revealed concentrations of amino acids and nicotinic acid averaging 0.35 mM and 0.56 microgram ml-1 respectively.

Amino Acids↗

Influence of detergent washing powders on minimal eliciting patch test concentrations of nickel and chromium.

Minimum eliciting levels of nickel have been estimated in 25 nickel-sensitive subjects, and of chromium in 14 chromium-sensitive subjects by patch tests with aqueous solutions of the respective metals. The minimum level of each metal required to provoke a patch test reaction was considerably greater than that found in fabric washing powder solutions and was in the majority of patients tested of the order of 112 ppm nickel (0.05% nickel sulphate) or 885 ppm hexavalent chromium (0.25% potassium dichromate). One nickel-sensitive subject and one chromium-sensitive subject reacted to 1 ppm of the respective metal. Fabric washing powder did not significantly alter the patch test reaction to nickel sulphate or provoke reactions in nickel- or chromium-sensitive subjects. EDTA significantly reduced the number and severity of patch test reactions to nickel sulphate but not those to potassium dichromate or trivalent chromium.

Chlorides↗

Absence of anamnestic response after transfusion of washed red blood cells in haemophilia A patients with antibody to factor VIII.

Washed red blood cells (WRC) have been used for replacing the blood loss in haemophilia A patients with antibody to Factor VIII. Levels of VIII coagulant activity (VIII:C), VIII coagulant antigen (VIII:CAg) and VIII related antigen (VIIIR:Ag) have been measured during the different steps of the preparation of WRC. The amount of VIII:CAg decreases very rapidly after washing and both are undetectable in the final product. These results were correlated with the absence of anamnestic response in 10 haemophilia A patients with inhibitor known as high responders.

Antibodies↗

Methanol exposure to car occupants from windshield washing fluid: a pilot study.

UNLABELLED: Automobile occupants might be exposed to considerable amounts of methanol from previously unreported source, namely via the inhalation of vapors of winter-grade, methanol-based, windshield washing fluid that drains to the intake air ducts of the car. Air samples were collected in passenger cars during simulated operating conditions and analyzed for methanol via headspace gas chromatography-mass spectrometry, electron impact, selected ion monitoring. The method was linear in the 2-2000 ppm range. Concentrations exceeding 1000 ppm were recorded. PRACTICAL IMPLICATIONS: Using a winter-grade, methanol-based, windshield washing fluid for windshield cleaning in a passenger car can result in a methanol concentration in the air of the passenger cabin in excess of 1000 ppm. In view of the widespread use of this product, more studies are necessary to elucidate, in depth, the concentrations of methanol vapors which could be encountered in various weather and driving conditions as well as the concomitant contributing influences of car design. These studies are necessary to properly assess the hazards associated with use of the fluid and possible mitigation approaches which might include substitution of methanol by less toxic formulations.

Air Pollutants↗

Cells and cellular interactions in gingival crevice washings from patients with juvenile periodontitis.

The cells and the cellular interactions in the gingival crevices of three patients with juvenile periodontitis (JP), three patients with adult periodontitis (AP) and one healthy control were studied using light and scanning electron microscopy (SEM). The cells were collected by the washing method. The washings were either 1) fixed for 30 min in 2.5% glutaraldehyde (GA), or 2) cytocentrifuged onto a Melinex-polyester film and fixed in GA. On the films the cells of interest were identified and light photomicrographs were taken before processing for SEM. Four types of cellular interactions were seen in SEM: between polymorphonuclear leukocytes (PMN) and epithelial cells, between PMNs and bacteria, between PMNs and between bacteria. Based on morphologic and structural interpretation, the PMNs in the local lesion in JP seemed 1) to have a different morphology from the PMNs of AP lesions, 2) to be capable of ingesting bacteria, 3) to accumulate in gingival pockets in numbers as high as in AP, and 4) to be overwhelmed by the challenge of bacteria and bacterial aggregates.

Adolescent↗

Proteolysis of human fixed, washed platelets by gram-negative bacterial metalloproteases: effect on von Willebrand factor-human platelet interactions.

Fixed, washed platelets (FWP) are usually stable to aggregation with von Willebrand factor (vWF) from human and certain animal plasmas over several months of storage. When one lot of FWP lost its stability in less than 1 week, studies demonstrated contamination with Serratia marcescens. Extracellular proteases produced by S. marcescens, as well as by Pseudomonas aeruginosa and Escherichia coli, were found to cause loss of FWP aggregability. Purified proteases were prepared from cell-free culture filtrates of S. marcescens and two different strains of P. aeruginosa. They were used to study the effect on the interaction of FWP and vWF. All three purified proteases destroyed FWP aggregability in a time- and concentration-dependent fashion. The protease produced by S. marcescens (SP) was found to be at least eight times more potent against FWP as a substrate than either of the two P. aeruginosa enzymes. The ability of SP to destroy FWP aggregability was prevented by EDTA and could be restored by the addition of Zn2+ in slight molar excess. When compared with trypsin and chymotrypsin, SP was found to be highly selective in digesting the FWP membrane, even at concentrations greater than that established to give a similar loss of FWP aggregability. SP does not induce aggregation of fresh, washed platelets or PRP, but renders them unaggregable with vWF. These proteases may be useful research tools for studying membranes and vWF-platelet interactions.

Blood Coagulation Factors↗

Settling of insecticide from dip wash mixed with dam water and zinc sulphate and used to control sheep lice (Bovicola ovis).

Insecticidal dipping fluid emulsions, mixed in vitro in dam water containing suspended clay particles and 1% w/v zinc sulphate, were analysed to determine rates of settling of diazinon, cyhalothrin and cypermethrin. Fifteen minutes after mixing, the concentration of the insecticides 5 cm below the surface had declined by 72.5%, 72.8% and 89.4%, respectively. On remixing, the concentration of insecticide in suspension was close to or greater than the initial concentration. In 2 trials, lice were eradicated from sheep showered with dip wash mixed in cloudy dam water to which 1% w/v of zinc sulphate was added. In 12 flock treatments in which 1000 to 2000 sheep were dipped with added zinc sulphate, the concentration of insecticide remained above the minimum lethal for susceptible strains of lice. However, lice were still present 6 months later in 8 of these flocks. When zinc sulphate is added to dip wash, agitation is needed to maintain the insecticide in suspension.

Animals↗

[Selective screening of enterohemorrhagic Escherichia coli depending on hemolysis on Beutin's washed sheep blood agar plates].

Recently, Beutin et al have studied hemolysin production in a large number of serologically diverse verotoxin (VT) producing Escherichia coli (VTEC) strains and found a close association between hemolysin production and VT production. This study was to examine whether hemolysin production on sheep blood agar can be used as an epidemiological marker for microbiological screening of enterohemorrhagic Escherichia coli (EHEC). In comparison of hemolysis on washed blood agar plates prepared in our laboratory and commercially available blood agar, all VT producing strains exhibited a typical hemolytic phenotype on the former, on the other hand, VT non-producers were less hemolytic on the former than the latter. On the washed blood agar, 11 VT1 producing strains belonging to 4 different serotypes, 1 strain of VT2 producer, 5 VT1&2 producers belonging to 2 different serotypes, all the 17 strains of VTEC were positive for hemolysis. The 85 VT non-producing strains belonging to 18 different serotypes isolated from faecal specimens from patients with diarrhea were tested, 10 strains (12%) belonging to 4 different serotypes (O6, 18, 26, UT: untypable) produced hemolysin. Two-hundred four (10%) of the 2,036 strains isolated from faecal specimens from 507 healthy persons exhibited typical hemolysis. As a result of identification of these 204 hemolytic strains, 148 strains of E. coli 13 strains of Citrobacter and 10 strains of other enterobacteria, and 13 strains of Pseudomonas, 12 strains of Aeromonas, 5 strains of Proteus and 3 strains of Vibrio were detected. 148 hemolytic strains of E. coli were carried out O-serotyping with commercially available pathogenic E. coli antisera, 57 strains (39%) were classified into 11 different serotypes, but 91 strains were not typable. Most frequent serotype was O18 (24 strains), next O6 (18 strains), and other 8 different serotypes (O25, 26, 28ac, 29, 55, 146, 152, 159) were less than 2 strain respectively.

Bacterial Toxins↗

Washing away your sins: threatened morality and physical cleansing.

Physical cleansing has been a focal element in religious ceremonies for thousands of years. The prevalence of this practice suggests a psychological association between bodily purity and moral purity. In three studies, we explored what we call the "Macbeth effect"-that is, a threat to one's moral purity induces the need to cleanse oneself. This effect revealed itself through an increased mental accessibility of cleansing-related concepts, a greater desire for cleansing products, and a greater likelihood of taking antiseptic wipes. Furthermore, we showed that physical cleansing alleviates the upsetting consequences of unethical behavior and reduces threats to one's moral self-image. Daily hygiene routines such as washing hands, as simple and benign as they might seem, can deliver a powerful antidote to threatened morality, enabling people to truly wash away their sins.

Emotions↗

Effect of ribosomal wash factors on inhibition by chloramphenicol.

In vitro protein-synthesizing systems from Escherichia coli can be categorized as either chloramphenicol-sensitive or chloramphenicol-insensitive. The chloramphenicol-sensitive systems used in this study required the presence of factors removed from ribosomes with 1.0 m NH(4)Cl when chromatographically purified ribosomes were used for amino acid incorporation. These ribosomal wash factors inhibited but did not eliminate amino acid incorporation in chloramphenicol-insensitive systems. For both systems, addition of increasing amounts of the ribosomal wash factors increased the sensitivity to chloramphenicol inhibition.

Amino Acids↗