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Flow cytometric study comparing paired bladder washing and voided urine for bladder cancer detection.

Paired bladder washings and voided urines from bladder cancer patients were compared as sources of exfoliated cells for detection of bladder carcinoma by flow cytometry (FCM). Bladder specimens fixed in 25% ethanol within sixty minutes of collection were found to be superior to unfixed bladder specimens. The percentage of specimens with good DNA resolution was greater for bladder washings (67% unfixed, 90% fixed) than voided urines (41% unfixed, 66% fixed). There was no difference in DNA resolution between specimens that remained unfixed less than one day, one day, or two days suggesting that the cells undergo the majority of degradation within a critical period soon after collection. Once fixed, there was no difference in DNA resolution for up to nineteen days, which suggests the feasibility of specimen transport to central FCM laboratories. Eighteen percent of unfixed bladder washings and 33 percent of unfixed voided urine specimens contained an insufficient number of cells (less than 5,000) at the time of analysis compared with 6 percent bladder washings and 17 percent voided urines fixed in 25% ethanol. Flow cytometry and cytology results were concordant in 28/43 (65%) of fixed bladder washings and 9/13 (69%) of fixed voided urine. Voided urine was unreliable in providing consistent FCM data due to the high number of specimens with poor resolution or insufficient cells and is not recommended as a substitute for bladder washing when screening high-risk populations or monitoring patients with past history of bladder cancer.

Carcinoma, Transitional Cell↗

Comparison of nasopharyngeal washings and swab specimens for diagnosis of respiratory syncytial virus by EIA, FAT, and cell culture.

Respiratory secretions for viral diagnosis are often collected with nasopharyngeal (NP) swabs, although many laboratories recommend NP aspirates or washings. We compared results using NP washings and NP swabs in three diagnostic RSV tests, a rapid RSV EIA antigen test (Abbott Laboratories), an indirect fluorescent antibody test (FAT) with rabbit antiserum, and virus culture (HEp-2 cells). Paired samples were collected from 121 children with suspected RSV bronchiolitis or pneumonia. A minitip swap was passed into the nasopharynx for 10 sec, rotated and withdrawn. The opposite nares was irrigated with approximately 1 ml of saline and aspirated using a syringe and plastic feeding tube. Fifty-one children (42%) grew RSV in culture, 49 from NP washings versus 27 from NP swabs (p less than 0.001). Fifty-three (44%) were positive by FAT, 52 from NP washings versus 12 from NP swabs (p less than 0.001). Fifty-eight children (48%) had positive RSV EIA tests, 57 from NP washings versus 35 from NP swabs (p less than 0.001). Detection by EIA was more sensitive than culture regardless of the method of specimen collection. We conclude that NP washings are superior to NP swabs for RSV culture and rapid diagnosis by EIA or FAT.

Child↗

Fluorescence in situ hybridization patterns in newly diagnosed superficial bladder lesions and corresponding bladder washings.

A multiprobe interphase fluorescence in situ hybridization (I-FISH) approach has become a useful ancillary tool in the follow-up protocol for patients with low-grade superficial bladder tumors. Nevertheless, reports contextually comparing I-FISH patterns in primary superficial tumor cells with those in concomitant washing cells at the time of initial tumor appearance are sparse. We comparatively evaluated I-FISH patterns of chromosomes 3, 7, 9, and 17 and of the CDKN2A and TP53 loci in newly diagnosed superficial bladder lesions and in corresponding bladder washings, to verify representatives of the latter type of sampling and to improve the efficacy of I-FISH follow-up. A total of 21 biopsies and 12 washings were examined. Samples obtained at the time of the tumor's first appearance showed the presence of cytogenetically abnormal clones in 80% of washings and 70% of biopsies. Five cases showed overlapping washing and biopsy I-FISH patterns; in three cases (and to a lesser extent in two others), consistent discrepancies between the two patterns was observed. The results indicate that knowledge of I-FISH patterns in both washing and biopsy cells on first tumor appearance may be of help in interpreting further follow-up I-FISH patterns, and that these should be considered in the context of the patient's entire clinical history.

Aged↗

Remediation of arsenic-contaminated soils and washing effluents.

Laboratory experiments were conducted to determine the distribution of various arsenic species in tailings and soils. Other specific goal of the tests were to evaluate the extraction efficiency of arsenic using alkaline or acid washing, to determine optimum operational parameters of alkaline washing, and to evaluate the arsenic precipitation of washing effluents by pH adjustment or ferric chloride addition. Alkaline washing using sodium hydroxide was found to be favorable in removing arsenic from tailings or soils having a higher portion of arsenic in the operationally defined crystalline mineral fraction of crystalline oxide and amorphous aluminosilicates. This is due to the ligand displacement reaction of hydroxyl ions with arsenic species and high pH conditions that can prevent readsorption of arsenic because predominant negatively charged crystalline oxides do not attract the negatively charged oxyanions. For tailings, sodium hydroxide had 10-20 times higher extraction efficiencies than hydrochloric- or citric acid. The optimum concentration of sodium hydroxide for soil washing was determined to be 200 mM for all samples, while the optimum ratios were 10:1 and 5:1 for tailings and field/river sedimentary soils, respectively. The washing effluent of river soil was effectively treated by adjusting pH to 5-6 with hydrochloric acid, resulting in arsenic concentrations of <50 microgl(-1). In the case of field soil effluent, an addition of ferric chloride with a minimum mass ratio of 11 (Fe/As) was needed to reduce the arsenic below 50 microgl(-1).

Alkalies↗

Removal of Pb and MDF from contaminated soils by EDTA- and SDS-enhanced washing.

Heavy metal- and organic-contaminated sites are ubiquitous, but few studies have been conducted to address such an issue. EDTA- and SDS-enhanced washing was studied for remediation of Pb- and/or marine diesel fuel (MDF)-contaminated soils. The feasibility of recovery and reuse of EDTA and SDS, as well as the physicochemical interactions among the chemical agents, contaminants and soils were extensively investigated using batch experiments. The optimal washing sequence was then determined. The experimental results showed that EDTA could be recovered and reused for four cycles without significant loss of its chelating capacity, while the extraction capability of SDS was noticeably reduced after each reuse cycle. The free phase of marine diesel fuel (MDF) in soils physically isolated the sorbed Pb on soils and thus reducing its extraction by EDTA. The presence of SDS alone or together with low concentration of EDTA was found to enhance Pb removal probably via electrostatic interaction and dissolution of soil organic matter. However, it hindered Pb extraction by high concentration of EDTA, because of the potential formation of complexes between some strongly-bound Pb and SDS, that are more resistant to desorption. Therefore, EDTA washing followed by SDS achieved the highest Pb removal efficiency. On the other hand, MDF removal by SDS was significantly hindered by coexisting Pb in soils, probably because the formation of Pb-dodecyl sulfate (DS) complex would decrease the effective amount of SDS available for forming micelles in solution and enhance MDF sorption. EDTA alone or together with SDS could enhance MDF removal, but the residual MDF after EDTA-washing became more resistant to SDS removal. Consequently, SDS washing followed by EDTA is considered as the optimal washing sequence for MDF removal.

Chromatography, High Pressure Liquid↗

Synchronous sperm retrieval and sperm washing in an intracytoplasmic sperm injection cycle in an azoospermic man who was positive for human immunodeficiency virus.

OBJECTIVE: To present the first reported case of synchronous sperm retrieval followed by sperm washing before an intracytoplasmic sperm injection (ICSI) cycle in an HIV-positive azoospermic man. DESIGN: Case report. SETTING: Assisted reproduction center. PATIENT(S): A 40-year-old HIV-positive man with obstructive azoospermia due to vasal aplasia. INTERVENTION(S): Synchronous sperm retrieval, sperm washing, nucleic acid-based sequence amplification testing, and intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Successful sperm retrieval sufficient for sperm washing and fertilization. RESULT(S): Sufficient quantity of spermatozoa for washing was obtained at epididymal aspiration. After the wash, HIV ribonucleic acid (RNA) was undetectable with nucleic acid-based sequence amplification testing, enabling injection of oocytes collected after routine gonadotropin superovulation. Of seven oocytes collected from the 39-year-old woman partner, six were injected and five fertilized (83%). Three embryos were transferred on day 2. The pregnancy test was negative on this occasion. CONCLUSION(S): This case demonstrates that sperm washing can be applied in cases of sperm retrieval where sperm volume and density is low, allowing the treatment of azoospermic HIV-positive men.

Adult↗

Effects of nonaqueous phase liquids on the washing of soil in the presence of nonionic surfactants.

The removal of malathion from soil by surfactant washing was investigated under various physical-chemical states of the malathion. Three distinctive phases (without nonaqueous phase liquids (NAPL), with NAPL, and the transitional zone of NAPL) were found to be important for a better understanding of the washing process. When there is no NAPL in the system, the washing process is less dependent on the surfactant dose if the applied surfactant concentration is above the critical micelle concentration. The existence of a sorption site boundary, which for the determination of different washing mechanisms, was identified. In the presence of NAPL, the washing performance is generally independent of the organic content (f(oc)) of the soils but is dominated by the concentration of the surfactant used, due to the lesser resistance for mass transfer in NAPL. If the formation of NAPL is marginal, a two-stage washing pattern is observed, which has been quantified by the term 'unit extraction'. For this two-stage system, a mathematical model was derived based on the observed initial unit extraction and final extraction capacity, which eventually resulted in a practical design equation with the use of primary parameters such as f(oc) and surfactant dose.

Detergents↗

Does diagnostic hysteroscopy in patients with stage I endometrial carcinoma cause positive peritoneal washings?

OBJECTIVES: Previous studies have shown that positive peritoneal washings may adversely affect cancer survival rates and that hysteroscopy is associated with a higher risk of positive washings in patients with endometrial carcinoma. Our aim was to assess if diagnostic hysteroscopy increases the risk of positive peritoneal washings in patients with endometrial cancer and affects the prognosis after surgery. STUDY DESIGN: Retrospective cohort study. The medical records of 50 consecutive patients with endometrial carcinoma, diagnosed with hysteroscopy and tissue sampling and treated by abdominal hysterectomy with bilateral salpingo-oophorectomy and peritoneal washings were reviewed. RESULTS: Of the 43 patients with endometrial carcinoma FIGO stage I, none had positive peritoneal washings (95%CI: 0-8.2%). The mean interval between hysteroscopy and surgery was 33.5 days. The 5-year disease-specific survival rate was 91.8%, the 5-year recurrence-free survival rate was 85.4%. CONCLUSIONS: Diagnostic hysteroscopy had no adverse effect on the incidence of positive peritoneal washings or on prognosis in stage I endometrial cancer patients.

Adult↗

In vitro assessment of the structural integrity of the female condom after multiple wash, dry, and re-lubrication cycles.

Since the introduction of the female condom in the early 90s, there have been numerous reports of reuse of the device. In response to these reports, studies were undertaken to evaluate the safety of female condom reuse. If reuse were shown to be safe, then programmatic costs of introduction of the female condom would be reduced allowing it to be more widely available. This article outlines the results of in vitro structural integrity testing of the female condom after multiple wash, dry, and re-lubrication cycles. Devices were tested up to 10 washes using water leakage, burst, and tensile seam testing. All results were compared to the United States Food and Drug Administration (US FDA) standards for an unused female condom. The results of the structural integrity tests for all 6 washing procedures examined in this study were above the FDA minimum standards for seam strength and burst tests. For the water leakage test, 3 of 6 washing procedures tested passed the required FDA minimum standards (no holes detected). From the results of the study, it seems that washing, drying, and re-lubricating the female condom up to 10 times leads to some deterioration in the structural integrity of the device for specified washing procedures. Further studies are currently being conducted to establish the safety of female condom reuse with respect to microbial retention, structural integrity after in vivo use, and viral permeability.

Single-Use Internal Condom↗

Sperm washing and swim-up technique using antibiotics removes microbes from human semen.

Pelvic infections may follow intrauterine or intratubal insemination with washed semen. In this study, we determined whether sperm washing removes microorganisms from human semen. Before and after semen wash, we cultured 15 ejaculates for aerobic and anaerobic bacteria, genital mycoplasma, and chlamydia. All semen samples had from one to five organisms isolated (total, 40 isolates) before the semen wash preparation. The mean number (+/- standard deviation) of isolates per sample was 2.67 +/- 1.35. After the semen were prepared, none of the samples showed a positive culture. The decrease in the number of samples with positive cultures and the decrease in the number of isolates is significant (P less than 0.0001). After sperm washing, electronmicroscopic studies did not show any microbes attached to any portion of the spermatozoa. We conclude that the method of sperm wash preparation used is effective in removing microbes present in human semen.

Anti-Bacterial Agents↗

Experience with pelvic washings in stage I and II endometrial carcinoma.

Pelvic washings for cytologic analysis have become an accepted diagnostic procedure in the management of endometrial carcinoma. A retrospective study was performed in 163 consecutive patients with FIGO Stage I and II endometrial carcinoma to assess the significance and value of the washings. Abnormal results were obtained in 5.5% of patients. Patients with FIGO grade 3 lesions were significantly most likely to have abnormal washings (P less than 0.05). Significant association was not observed with stage, depth of myometrial invasion, extrauterine spread, histologic subtype, prior hormone usage, or hormone receptor status. Multivariant analysis indicated that, for Stages I and II combined and for Stage I alone, FIGO grade 3 was most predictive of poor survival (P less than 0.01). The postoperative therapy of only 2 patients (1.2%) was altered by the results of the pelvic washings and only one of these patients has survived. No patients have had recurrent disease (median follow-up 31 months) in the peritoneal cavity with negative washings unless two or more other adverse prognostic factors were present. It is concluded that pelvic washings have a limited role in the clinical management of Stage I and II endometrial carcinoma.

Carcinoma↗

The recovery of ureaplasmas from bovine embryos following in vitro exposure and ten washes.

Twenty-six unhatched embryos and ova were exposed to Ureaplasma diversum strain 2312 in vitro for 16 h and subsequently washed ten times. Fifteen of the embryos and their wash fluids were cultured for ureaplasmas. Of the remaining 11 embryos, six were incubated with rabbit anti-Ureaplasma immunoglobulin (RAI) and five were incubated with serum from naive rabbits(NRS), after which all were incubated with protein A gold and prepared for electron microscopy. On ultrastructural examination, ureaplasmas were observed on the outer surface of the zona pellucida of all 11 embryos. The ureaplasmas on the six embryos incubated with RAI were labeled with gold particles, while those on the five embryos incubated with NRS were not labeled. Ureaplasmas were recovered from all 15 of the cultured embryos and all of the first and second wash fluids as well as intermittently from the third, fourth, sixth, seventh, eighth and ninth wash but not fom the fifth or tenth wash. It was concluded that viable ureaplasmas adhered to the zona pellucida during in vitro exposure of bovine embryos and were not removed by ten washes.

Journal Article↗

A comparative study of telomerase activity in sputum, bronchial washing and biopsy specimens of lung cancer.

The potential of telomerase, the ribonucleoprotein enzyme, as a non-invasive screening marker was studied in pre-bronchoscopy sputum (S), bronchial washings (W) and bronchoscopic biopsy (B) samples from individuals under evaluation for lung cancer. Out of the 52 cases studied, 42 were clinically suspected primary lung cancer patients and 10 had pulmonary disorders but had no clinical evidence of lung cancer. Fifteen (39.5%) S samples, 24 (63.1%) W samples and 32 (84.2%) B samples, which were cytologically/histopathologically positive were also positive for telomerase activity. Interestingly, 16 (42%) S samples, 20 (52.6%) W samples and 20 (52.6%) B samples, initially reported cytologically/histopathologically negative, showed detectable telomerase activity. Lung cancer was finally confirmed in these cases by repeat cytology/histopathology. However, telomerase activity was detected in 31 (81.6%) S, 26 (68.4%) W and 33 (86.8%) B samples of suspected lung cancer patients. Telomerase activity was negative in S, W, and B of four of the suspected cases, which ultimately turned out to be negative for lung cancer. Cytopathology/histopathology alone (including repeat attempts) identified 15 (39.5%) cases of sputum, 24 (63.1%) cases of bronchial washings and 32 (84.2%) bronchoscopic biopsy samples. Out of 10 controls, low telomerase activity was detected in only one (10%) of the bronchial washings, which later turned out to be due to large number of inflammatory cells. Telomerase activity assay of sputum carried sensitivity, specificity and diagnostic accuracy of 81.6, 100 and 86.5%, respectively, while that for bronchial washing was 68.4, 100 and 76.9%, respectively, and for bronchoscopic biopsy samples was 86.8, 100 and 88.1%, respectively. A positive correlation (P<0.01) was seen between age and telomerase activity in sputum, bronchial washing and biopsy samples but no significant correlation was seen between sex and telomerase activity or duration of smoking and telomerase activity. A significant positive correlation was observed between staging and telomerase activity in sputum (P<0.01), bronchial washing (P<0.01) and biopsy samples (P<0.01). Our findings indicate that telomerase is a specific marker for malignant lung disease and can complement cytology/histopathology in the diagnosis of lung cancer. Sputum telomerase assay holds the potential for early and non-invasive diagnosis of lung cancer.

Adult↗

Comparison of cloth, paper, and warm air drying in eliminating viruses and bacteria from washed hands.

We compared the efficiency of paper, cloth, and electric warm air drying in eliminating rotaviruses and Escherichia coli remaining on finger pads washed with 70% isopropanol, a medicated liquid soap, an unmedicated liquid soap, or tap water alone. The contaminated area on the finger pads of a volunteer was exposed to the hand-washing agent for 10 seconds and then rinsed in 40 degrees C tap water. The washed areas were dried for 10 seconds by one of the three methods. Irrespective of the hand-washing agent used, electric air drying produced the highest and cloth drying the lowest reduction in the numbers of both test organisms. These findings indicate the importance of selecting the right means for drying washed hands, particularly when less effective hand-washing agents are used.

Disinfectants↗

Levels of naturally occurring microorganisms on surgical instruments after clinical use and after washing.

Surgical instruments exposed to sterile body sites should be contaminated with relatively low levels of microbial contamination or bioburden; however, few studies in the literature have determined the quantitative level and types of contamination. A study was conducted at 2 clinical sites to determine the level of microbial contamination of surgical instruments after clinical use and after washing. Quantitative assays showed that bioburden levels were in the range of 0 to 4415 colony forming units per instrument after clinical use, and 88% of the instruments had bioburden levels lower than 1000. As expected, a reduction in counts occurred after washing; however, in some cases, higher counts were found on the instruments after the washing process. Although the washing procedure is effective in reducing the microbial levels deposited on the surgical instruments during use, a recontamination process occurs that results in increased counts after washing. The low bioburden level after washing consists of predominantly vegetative microorganisms that present a relatively low challenge to sterilization and disinfection systems.

Colony Count, Microbial↗

Effect of teat washing on the bacteriological contamination of milk from cows kept under various housing conditions.

The effect of different methods of teat washing on bacterial contamination and sediment levels of the milk of cows kept under 3 housing conditions was studied. By use of an in line milk sampler and steam sterilized clusters the bacterial contamination from the teats after udder preparation was determined directly. The results show that, even under the same housing conditions and teat washing treatments, considerable variation in bacterial counts and sediment level occurred. Overall, washing with hypochlorite (600 mg/l available Cl) and drying with a paper towel gave total counts that were significantly lower than were obtained with the other 4 treatments. The aerobic spore count, in which thermoduric flora predominated, was greatly reduced by drying the teats, but the inclusion of hypochlorite in the wash water had little effect. Coliform contamination was very low (approximately 1/ml) even where the cows' teats were heavily soiled and were not washed. The sediment levels in the milk again showed wide variation and occasional high values (3 mg/l) were recorded for washed cows. In marked contrast to its effect on the bacteriological results, drying had little effect on reducing the sediment.

Administration, Topical↗

Failure of hand disinfection with frequent hand washing: a need for prolonged field studies.

In a prolonged field trial a 4% chlorhexidine digluconate detergent scrub (Hibiscrub(R)), that had earlier proved to be an effective hand disinfectant, was studied in hospital wards. Finger tips were found to harbour more bacteria than the hand dorsum and the samples collected from them yielded more information on the bacteriological and dermatological effects of hand disinfectants in practice.In wards with a relatively low hand-washing frequency (less than 20 times in 8 hours) the bacteriological results resembled those obtained by in-use tests with volunteers. In the neonatal unit where the hand washing frequency was remarkably high, even occasionally over 100 times/8 h shift, an increase in the bacterial colony counts of the majority of the staff was recorded both before and after hand washing already after using the preparation for 1 week. Age, occupation and hand-washing frequency all correlated with the bacteriological results. Twenty-seven out of 37 persons complained of side effects such as wounds of finger tips and redness or heavy drying of the skin. Wounds, particularly on finger tips, resulted in the failure of disinfection. An increase in bacterial counts was sometimes noted without any dermatological or subjective changes. Drying of the skin was complained of less often when no increase in skin bacteria occurred.After the changeover of washing practice to a detergent followed by a rinse with spirit solution containing chlorhexidine and glycerol a decrease was recorded in the bacterial counts. It is concluded that more attention should be paid to long-term testing of hand washing and disinfection methods to ensure optimum final results in practice. It is obvious that the knowledge obtained from short time in-use testing cannot be applied to all conditions of use.

Adult↗

Effectiveness of hand washing and disinfection methods in removing transient bacteria after patient nursing.

The effectiveness of various hand washing and disinfection methods in removing transient skin bacteria was studied in hospital after dry or moist contamination of the hands when nursing burn patients. The results were compared with those of laboratory tests with volunteers. A fairly good correlation of the bacterial reductions existed between hospital and laboratory tests. All other methods removed Staph. aureus from the hands more effectively than liquid soap. Gram-negative bacilli were more easily removed than staphylococci, even with soap wash alone. In hospital, none of the washing and disinfection methods always removed all patient-borne bacteria from the hands. After dry or moist contamination and subsequent washing with soap only, colonies of Staph. aureus were often detected in finger-print samples. Staphylococci were more often completely removed by a 4% chlorhexidine detergent scrub and alcoholic solutions (either with or without previous soap wash) than by liquid soap, hexachlorophene or iodophor preparations. Gram-negative bacilli were more easily removed by all the washing and disinfection methods. After moist contamination, Gram-negative bacilli were more often completely removed from the hands by ethanol than by other treatments. The results of the present study emphasize the importance of always using gloves when nursing a profuse spreader of bacteria or one who must be protected from infection.

Chlorhexidine↗