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A clinically applicable method to preserve urine and bladder washing cells for flow cytometric monitoring of bladder cancer patients.

We describe a method to fix exfoliated bladder cells that is suitable for followup of bladder cancer patients by deoxyribonucleic acid flow cytometry. After fixation with room temperature methanol plus acetic acid (20:1, volume:volume) urine and bladder washing samples from these patients can be stored at room temperature for 3 to 7 days and then assessed reliably for the presence of aneuploidy and the percentage of hyperdiploid cells. For those with active transitional cell carcinoma diagnostic accuracy comparing fresh to fixed specimens was improved from 58 to 92% with urine and from 50 to 100% with washing samples. For patients with a history of transitional cell carcinoma who currently are free of disease the false positive rate remains unchanged after fixation. The procedure described is suitable for use in the outpatient clinic and should permit shipping of samples without refrigeration to a central flow cytometry facility for analysis.

Carcinoma, Transitional Cell↗

Assessment of the cervix after acetic acid wash: inter-rater agreement using photographs.

OBJECTIVE: To estimate the inter-rater agreement on the assessment of cervical photographs after acetic acid wash, using visual inspection with acetic acid categories. METHODS: Three raters individually assessed 144 photographs as negative, positive, or suspicious for cancer. The inter-rater agreement was analyzed using the unweighted and weighted kappa coefficient. To explore the reasons for concordancy and discordancy, photographs were compared on histologic evidence of cervical intraepithelial neoplasia and on testing for oncogenic types of human papillomavirus. RESULTS: Overall raw agreement among the three raters was 66.7% (96 of 144) with a kappa of 0.57 (95% confidence interval 0.48, 0.66). Pair-wise agreement using unweighted and weighted kappa was moderate to substantial: 0.54-0.60 and 0.56-0.63, respectively. There was concordance on negative in 25.7% (37 of 144) and on positive or suspicious for cancer in 41.0% (59 of 144). Cervical intraepithelial neoplasia II or III was not present on biopsy if photographs were concordant-negative, and the human papillomavirus test was less likely to be positive (relative risk 0.3; 95% confidence interval 0.2, 0.6) in concordant-negatives compared with concordant-positives, including suspicious for cancer. Cervical intraepithelial neoplasia II or III was more common in photographs that were concordant-positive, including suspicious for cancer, compared with discordants (relative risk 3.4, 95% confidence interval 1.5, 7.6). CONCLUSION: Based on photographs of the cervix taken after acetic acid wash, the level of agreement among raters using visual inspection with acetic acid categories was moderate to substantial, consistent with other commonly used tests.

Acetic Acid↗

Washing powders and the environment: has TAED any influence on the complexing behaviour of phosphonic acids?

Complexation properties of two phosphonic acids: nitrilotris (methylenephosphonic acid) and 1-hydroxyethane-1,1'-diphosphonic acid, with Cu(II) and Ca(II) have been previously determined by pH and ionic selective electrode titrations. These phosphonic acids are commonly added to washing powders as polyphosphate substitutes and are here studied in the presence of another detergent compound: tetraacetylethylenediamine (TAED), a bleaching activator. Product concentrations were chosen in order to correspond to component concentration ratios encountered in washing powders. Potentiometric titrations were carried out to determine the possible TAED interferences; they indicate that TAED and its by-products have no action on phosphonic acid complexing behaviour. Under our experimental conditions, the action of TAED was modelled with acetic acid and ethylenediamine, the final by-products of TAED hydrolysis. If we take into account both phosphonic and acetic acids, speciation diagrams corresponding to representative fresh water systems showed that the acetic acid does not influence cation speciation.

Copper↗

The Feulgen reaction: importance of the wash after hydrolysis.

Washing in absolute ethanol after FEULGEN hydrolysis minimizes losses of DNA. The hydrolysis in HCl diluted in tetrahydrofuran is not superior in this respect to conventional hydrolysis if this is followed by an ethanol wash. These conclusions are based on cytophotometric and radiometric experiments made on salivary glands of sciarid larvae.

Animals↗

Treatment with hypertonic saline versus normal saline nasal wash of pediatric chronic sinusitis.

BACKGROUND: Chronic sinusitis (CS) is a common disease in children, especially those with allergies, that is caused by impaired drainage from the sinuses. Hypertonic NaCl solution has been shown to increase mucociliary clearance and ciliary beat frequency. OBJECTIVE: We performed a randomized double blind study to compare the effect of nasal wash with hypertonic saline (HS) (3.5%) versus normal saline (NS) (0.9%) on CS. METHODS: Thirty patients with CS aged 3 to 16 years were studied. They were randomly divided into two treatment groups matched by age and severity of the disease. Each individual was treated with either HS or NS for 4 weeks. All patients were evaluated by two clinical scores (cough and nasal secretions/postnasal drip [PND]) and by a radiology score at the beginning of the study and after 4 weeks. RESULTS: The HS group improved significantly in all scores (average +/- SD): cough score, from 3.6 +/- 0.51 to 1.6 +/- 0.74; nasal secretion/PND score, from 2.86 +/- 0.35 to 1.6 +/- 0.74; and radiology score, from 8.06 +/- 1.28 to 2.66 +/- 1.04. The NS treatment group showed significant improvement only in the PND score (from 2.66 +/- 0.49 to 1.53 +/- 0.83) but no significant change in both the cough score (from 3.53 +/- 0.52 to 3.33 +/- 0.49) and the radiology score (from 8.13 +/- 1.25 to 7.86 +/- 0.91). Clinical observation 1 month after the end of the study showed no change compared with the end of the study in both groups. CONCLUSION: HS nasal wash is an efficient treatment of CS.

Adolescent↗

IL-4 and IL-5 mRNA expression in induced sputum of asthmatic subjects: comparison with bronchial wash.

BACKGROUND: The local production of TH2 -type cytokines is thought to orchestrate the ongoing eosinophilic inflammation and contribute to the pathophysiologic features of allergic asthma. Previous studies investigating cytokine expression in asthmatic individuals have used invasive fiberoptic bronchoscopy techniques. To date, there have been no reports of cytokine mRNA expression in induced sputum as a means of quantifying local inflammatory events. OBJECTIVES: We examined whether IL-4, IL-5, and IFN-gamma mRNA expression could be detected in cells from induced sputum in subjects with mild asthma and normal control subjects. In addition, we compared the profile of inflammatory cells and cytokine mRNA in sputum and bronchial wash fluid. METHODS: Cells positive for IL-4, IL-5, and IFN-gamma mRNA were determined by using in situ hybridization on cytospun aliquots of sputum induced by successive inhalations of hypertonic saline. Inflammatory cells were quantified by using immunologic cell surface markers and immunocytochemistry. RESULTS: IL-4 and IL-5 mRNA were detected in the sputum of all asthmatic subjects, and the number of cells expressing these cytokines was significantly higher than that found in control subjects. Colocalization studies showed CD3-positive T cells were the major sources of IL-4 and IL-5 mRNA. CONCLUSIONS: This study demonstrates that induced sputum can be used to detect mRNA for TH2 -type cytokines in bronchial asthma and that the increase in IL-4 and IL-5 mRNA expression is similar to that seen with more invasive techniques. The qualitative differences in inflammatory cell numbers between sputum induction and bronchial wash are consistent with their sampling of different airway compartments.

Adult↗

New enzyme immunoassays for measurement of influenza A/Victoria/3/75 virus in nasal washes.

Nasopharyngeal (NP) washes from 12 patients experimentally infected with influenza A/Victoria/3/75 wild-type virus were tested in 2 newly developed enzyme-immunoassay (EIA) systems. Results were compared with infectivity titres of the NP wash specimens in rhesus monkey kidney-tissue culture. EIA detected A/Victoria/3/75 haemagglutinin antigen for more days than did culture. EIA is also rapid and easy to do in large numbers; hence it may have advantages over culture methods for the diagnosis of influenza infections and the investigation of influenza epidemiology.

Animals↗

Surface hydrophobic amino acid residues in cellulase molecules as a structural factor responsible for their high denim-washing performance.

The denim-washing performance of six purified fungal cellulases (four endo-1,4-beta-D-glucanases and two cellobiohydrolases) was compared using a model microassay. The performance of cellobiohydrolases per mg of protein was much lower than that of endoglucanases. For endoglucanases, it varied up to 5 times between the best and the worst enzyme. Experiments with amino acids immobilized on cross-linked agarose showed that their side chains may bind indigo owing to hydrophobic interactions and formation of hydrogen bonds. The best binding effects provided Tyr and Phe. Analysis of three-dimensional structures of cellulase molecules showed that a certain correlation exists between the washing performance of enzyme and (i) quantity (percentage) of aromatic residues exposed to solvent on the surface of protein globule or (ii) overall percentage of the surface hydrophobic residues. Data presented provide an evidence that the molecules of certain cellulases, which have hydrophobic domains (clusters of closely located non-polar residues) on their surface, may bind indigo and thus act as emulsifiers helping the dye to float out of cellulose fibers to the bulk solution.

Journal Article↗

Washing out of lipophilic compounds induces a transient increase in the passive Ca(2+) leak in permeabilized A7r5 cells.

We have investigated how the immunosuppressant drug FK506 affected the basal Ca(2+) leak in permeabilized A7r5 cells. Non-mitochondrial Ca(2+) stores loaded to steady state with Ca(2+) slowly lost their accumulated Ca(2+) during incubation in a Ca(2+)-free efflux medium. FK506 up to 100 microM had no effect on the basal Ca(2+) leak. In contrast, the rate of Ca(2+) release proceeded much faster immediately after washing out FK506. The increase in rate of Ca(2+) release after washing out of this compound depended on both its initial concentration and on the time of pre-incubation. A similar effect was also observed after removing another immunosuppressant drug (rapamycin) and after removing the inositol 1,4,5-trisphosphate receptor inhibitor xestospongin C. Since all these substances have a high octanol/H(2)O partition coefficient and accumulate in the endoplasmic reticulum membrane, we suggest that the transient increase in the basal Ca(2+) leak is due to the sudden removal of these lipophilic substances from the membrane.

Animals↗

Bacterial resistance and topical antimicrobial wash products.

Current scientific evidence has not shown that a link exists between the use of topical antimicrobial formulations and antiseptic or antibiotic resistance. As a result of the extensive history and varied use of antiseptic products and ingredients, any selective pressure for antibiotic resistance that may be occurring or may be uncovered in the future because of antiseptic use would be expected to be insignificant compared with the selective pressure because of antibiotic use. This review illustrates the effectiveness of topical antimicrobial wash products against antibiotic-resistant and antiseptic-resistant bacteria in use settings as well as the studies performed (antiseptic, deodorant, and oral care) demonstrating the lack of development of resistance in long-term clinical studies. Although these studies illustrate that the use of topical antimicrobial products have not been shown to play a role in the fluctuations of the specific composition or resistance of the skin flora, changes in skin flora have been shown to occur. Based on current knowledge, the benefit from use of topical antimicrobial wash products in combination with standard infection control and personal hygiene practices far outweighs the risk of increased antibiotic resistance.

Anti-Infective Agents, Local↗

Changes in concentration of pesticide residues in potatoes during washing and home preparation.

Monitoring of pesticide residues in potato tubers and their prepared products ("pommes frites" and chips) was undertaken. Experiments were carried out to determine changes in concentration due to the washing, peeling and cooking process (blanching and frying) to assess the stability of pesticides in potatoes and their products. Pesticide residues were quantified by using gas chromatography. Results show that malathion, HCB, lindane and p,p-DDD were predominant in potatoes and their products. The highest mean was detected in potatoes, followed by pommes frites, while the lowest mean was recorded in chips. On the other hand, potato skin samples were found to contain the highest levels of DDT and its derivatives, lindane and HCB. Peeling was necessary to remove the greatest amount of pesticides in the skin. Washing with water and/or other solutions as well as the cooking process (blanching and frying) helped to eliminate most of the pesticide residues from the potato tubers.

Chromatography, Gas↗

Particle size separation via soil washing to obtain volume reduction.

A pilot-plant study was performed using a soil washing pilot plant originally designed by the Environmental Protection Agency (EPA) to demonstrate scale-up and potential full-scale remediation. This pilot plant named VORCE (Volume Reduction/Chemical Extraction) was modified to meet the specific requirements for treatment of the Formerly Utilized Sites Remedial Action Program (FUSRAP) and a Department of Energy site soils. After a series of tests on clean soils to develop operating parameters and system performance, the machine was used to treat soils, one contaminated with Thorium-232 and the other with Cesium-137. All indicate that soil washing is very promising for volume reduction treatment. In addition, cost data was generated and is given herein.

Cesium Radioisotopes↗

The value of bronchoalveolar lavage and bronchial washings in the diagnosis of invasive pulmonary aspergillosis.

The objective of this study was to clarify conflicting reports of the sensitivity and specificity of bronchoalveolar lavage or bronchial washings for diagnosing invasive pulmonary aspergillosis. The study was a retrospective review of 300 consecutive patients in a tertiary referral centre subjected to 343 fiberoptic bronchoscopic procedures for the evaluation of pulmonary infiltrates. Classification of paired fungal culture and cytologic examination of bronchoalveolar lavage or bronchial washing fluid according to clinical, radiographic, histological and autopsy evidence of invasive pulmonary aspergillosis. One-hundred and fifteen deaths occurred, with a 58% autopsy rate. A diagnosis of invasive pulmonary aspergillosis was made in 21 immunosuppressed patients with 16 deaths. Bronchoalveolar lavage cytology showed aspergillus in 19 specimens (invasive pulmonary aspergillosis in 16), cultures yielded aspergillus in 41 (invasive pulmonary aspergillosis in ten), with both tests positive in nine. Cytology sensitivity was 64.0%, specificity 99.1%, positive predictive value 84.2%, and negative predictive value 97.2%. Culture sensitivity was 40.0%, specificity 90.3%, positive predictive value 24.4%, and negative predictive value 95.0%. Concordant cytology and culture sensitivity was 32.0%, specificity 99.7%, positive predictive value 88.9%, and negative predictive value 94.9%. In conclusion, when characteristic hyphae are visualized in bronchoalveolar lavage specimens from immunosuppressed patients with compatible clinical data, it is advisable to treat for presumptive invasive pulmonary aspergillosis.

Adult↗

A case of acute gasoline intoxication at the scene of washing a petrol tank.

We encountered a case of acute gasoline intoxication at the scene of washing the inner wall of a petrol tank. The decedent was a 50-year-old male, who was the supervisor. Two young workers wearing mask respirators began to wash the inner wall of the gasoline tank under poor ventilation. About 1 h later, because one of the workers lost consciousness, the supervisor entered the tank, without a mask respirator, to rescue the worker. However, the supervisor immediately fainted, and died 26 h after the accident. In the autopsy, blisters were observed on the skin of the face, neck, anterior chest, upper extremities, and back. The heart contained hemolyzed blood. Histologically, hemorrhagic pulmonary edema, pneumonia, and proximal tubular necrosis were found. In the toxicological analyzes, toluene, xylene, and trimethylbenzene were detected in the blood, brain, and gastric contents. Toluene concentrations in the blood and brain were 0.3 mug/ml and 3.7 mug/g, respectively. Since pathological findings were consistent with the reported findings concerning gasoline intoxication, and constituents of gasoline were in the body, death was attributed to acute gasoline intoxication. It was considered that sufficient ventilation and proper use of a mask respirator were essential for this kind of work.

Accidents, Occupational↗

Hair analysis for opiates: evaluation of washing and incubation procedures.

Hair analysis of drugs of abuse has been a subject of interest from a clinical, social and forensic perspective for years because of the broad time detection window after intake in comparison to urine or blood. However, the correct and reliable interpretation of opiates findings in an authentic hair sample requires optimalisation and standardisation of decontamination and incubation procedures. Comparing various published methods, we have found some variability in them and no unequivocal recommended procedure for starting with a method directly. Therefore, various combinations of solvents, of various polarity, as washing solvents were tested for removing opiates from the external surface of real hair samples. The yields of opiates from these washings were compared with the yields from the interior of the hair matrix after digestion with various procedures. The opiates after digestion were cleaned up from resulting solution on extraction columns with mixed solid-phase and analysed by GC-MS in standard EI mode after silylation. The efficiencies of neutral (Söerensen buffer, pH 7.4), acid (0.1 M HCl) and basic (1 M NaOH) digestion of the hair matrix were evaluated and the relative recoveries for morphine, codeine, dihydrocodeine and hydrocodone were compared. As it is very problematic to imitate the reference hair sample with a specific amount of analytes incorporated inside, which can be used for calibration to get a close estimate of the quantities of analytes inside the solid authentic sample, the total digestion of a hair sample in basic medium was considered to be a very important reference basis for quantitative determinations. The ratios of hydrolysis of labile 6-acetylmorphine or acetylcodeine were tested and evaluated in practical routine conditions of acid or neutral digestion of hair. Comparing the three methods of incubation of authentic hair samples, the methods using 1 M NaOH or 0.1 M HCl yielded higher recoveries of total equivalents of morphine or codeine, whereas the incubation in Söerensen buffer allowed the reflection of real ratios of labile metabolites and/or parent compounds in an original sample. This method has been shown to be capable of detecting hydrocodone in hair with other opiates concomitantly and to indicate the drug abuse pattern of a person at various time intervals in the past.

Gas Chromatography-Mass Spectrometry↗

Effect of a disinfectant udder wash and a post-milking teat dip on the bacterial population of the teat end and on the rate of new intramammary infection.

A pre-milking wash disinfectant procedure employing a 2% (w/v) chlorhexidine solution in a detergent basic did not reduce the rate of new intramammary infection in a lactating herd challenged with Staphylococcus aureus. Teat-end swabs showed no significant difference in the number of Staph. aureus isolated from the teat-ends pre-washed with either water, or water and chlorhexidine. In contrast, disinfecting teats after milking with a solution containing 5000 mg available iodine/l significantly reduced the staphylococcal population of the teat-ends and the rate of new intramammary infection, compared with teats not disinfected.

Animals↗

A differential neural response in obsessive-compulsive disorder patients with washing compared with checking symptoms to disgust.

BACKGROUND: Patients with obsessive-compulsive disorder (OCD) have symptoms that predominantly concern washing (washers) or checking (checkers), or both. Functional neuroimaging has been used to identify the neural correlates of the urge to ritualize but has not distinguished between washing and checking symptoms in OCD. We used functional magnetic resonance imaging to compare the neural response to emotive pictures in washers and checkers. METHODS: In one of two 5-minute experiments, washers (N = 7), checkers (N = 7) and age-matched normal controls (N = 14) were scanned while viewing alternating blocks of normally disgusting (rated as disgusting by all subjects) and neutral pictures. In the other experiment, all patients and a normal subgroup (N = 8) viewed alternating blocks of washer-relevant (rated as more disgusting by washers than normal controls or checkers) and neutral pictures. RESULTS: In all subjects, normally disgusting pictures activated visual regions implicated in perception of aversive stimuli and the insula, important in disgust perception. Only in washers were similar regions activated by washer-relevant pictures. In checkers, these pictures activated fronto-striatal regions associated with the urge to ritualize in OCD. Normal controls were more similar in neural response to checkers than washers to these pictures. Both normal controls and checkers had frontal regions activated significantly more by washer-relevant than normally disgusting pictures, and had these regions activated significantly more than washers by washer-relevant pictures. CONCLUSIONS: We demonstrate a differential neural response to washer-relevant disgust in washers and checkers: only washers demonstrate a neural response to washer-relevant disgust associated with emotion perception rather than attention to non-emotive visual detail.

Adolescent↗

Washing with contaminated bar soap is unlikely to transfer bacteria.

Recent reports of the isolation of microorganisms from used soap bars have raised the concern that bacteria may be transferred from contaminated soap bars during handwashing. Since only one study addressing this question has been published, we developed an additional procedure to test this concern. In our new method prewashed and softened commercial deodorant soap bars (0.8% triclocarban) not active against Gram-negative bacteria were inoculated with Escherichia coli and Pseudomonas aeruginosa to give mean total survival levels of 4.4 X 10(5) c.f.u. per bar which was 70-fold higher than those reported on used soap bars. Sixteen panelists were instructed to wash with the inoculated bars using their normal handwashing procedure. After washing, none of the 16 panelists had detectable levels of either test bacterium on their hands. Thus, the results obtained using our new method were in complete agreement with those obtained with the previously published method even though the two methods differ in a number of procedural aspects. These findings, along with other published reports, show that little hazard exists in routine handwashing with previously used soap bars and support the frequent use of soap and water for handwashing to prevent the spread of disease.

Adult↗