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PubMed1985

A simple and practical technic for detecting cancer cells in urine and urinary bladder washings by flow cytometry.

DNA measurement by flow cytometry has been demonstrated to be a potentially useful technic in the diagnosis of bladder cancer by detecting neoplastic cells in bladder washings and urine specimens. The authors' goal was to develop a simple and practical method utilizing the new generation of cytofluorographs designed for use in the clinical laboratory. This method combined direct fixation with cell lysis yielding fixed intact nuclei. Following RNase and pepsin digestion, the nuclei were separated from debris and aggregates on a sucrose barrier, stained with ethidium bromide, and analyzed with an argon laser analytic cytofluorograph. Urines and bladder washings from 14 patients with positive urinary cytology and histologically diagnosed bladder cancers were compared with specimens from patients without urothelial malignancies. DNA histograms clearly delineated aneuploid from diploid populations and often identified S, G2M, and G1 phase nuclei. Aneuploid populations have been detected in all tumor specimens with positive cytologies studied to date.

Aged↗

PubMed1990

32P-postlabeling assay for carcinogen-DNA adducts: description of beta shielding apparatus and semi-automatic spotting and washing devices that facilitate the handling of multiple samples.

The utilization of the 32P-postlabeling assay in combination with TLC for the sensitive detection and estimation of aromatic DNA adducts has been increasing in the past few years. The procedure consists of 32P-labeling of carcinogen-adducted 3'-nucleotides in the DNA digests using [gamma-32P]ATP and polynucleotide kinase, separation of 32P-labeled adducts by TLC, and their detection by autoradiography. During both 32P-labeling and initial phases of TLC, a relatively high amount of [gamma-32P]ATP (3.0-4.5 mCi) is handled when 30 samples are processed simultaneously. We describe the design of acrylic shielding apparatus, semi-automatic TLC spotting devices, and devices for development and washing of multiple TLC plates, which not only provide substantial protection from exposure to 32P beta radiation, but also allow quick and easy handling of a large number of samples, thus expediting the assay workup and making it less labor-intensive. Specifically, the equipment includes: (i) a multi-tube carousel rack (7.5 cm diameter and 7.7 cm height) having 15 wells to hold capless Eppendorf tubes (0.5 ml) and a rotatable lid with an aperture to access individual tubes; (ii) a pipet shielder; (iii) two semi-automatic spotting devices to apply radioactive solutions to TLC plates; (iv) a multi-plate holder for TLC plates; and (v) a mechanical device for washing multiple TLC plates. Item (i) is small enough to be held in one-hand, vortexed, and centrifuged to mix the solutions in each tube while beta radiation is shielded. Items (iii) to (iv) aid in the automation of the assay.

Carcinogenicity Tests↗

PubMed2004

The effect of human immunodeficiency virus on sperm parameters and the outcome of intrauterine insemination following sperm washing.

BACKGROUND: This is the first study to assess the outcome of sperm washing and intrauterine insemination (IUI) cycles in human immunodeficiency virus-positive (HIV(+)) men to determine any predictors of success, as well as evaluating the effect of HIV on sperm parameters. METHODS: Semen characteristics were evaluated in 106 HIV(+) men and a control group of 234 HIV(-) men, and the effect of markers of HIV disease assessed. Age, stimulation regime, sperm parameters, markers of HIV disease and the use of anti-retrovirals were assessed as predictors of the outcome of sperm washing/IUI cycles in the HIV(+) men. RESULTS: Ejaculate volume, sperm concentration, total count, progressive motility and normal morphology were all significantly higher in the control group compared to the HIV(+) men (P<0.05). A significant positive correlation was observed between CD4 count and sperm concentration, total count, motility, progressive motility type 'a'+'b' and post-preparation concentration and a significant negative correlation with normal sperm morphology of both raw and post-preparation samples. No correlation was observed between viral load (VL), years since diagnosis, use of anti-retrovirals or duration of use and any sperm parameter. The only factors that significantly improved IUI outcome were a VL <1000 copies/ml and the use of anti-retrovirals. CONCLUSIONS: These data demonstrate that sperm parameters are significantly impaired by the presence of HIV infection and in particular correlate with CD4 count. Undetectable VL and the use of anti-retrovirals improve the outcome of IUI/sperm washing in HIV(+) men.

Acquired Immunodeficiency Syndrome↗

PubMed2006

Detection of human immunodeficiency virus-1 RNA and DNA by extractive and in situ PCR in unprocessed semen and seminal fractions isolated by semen-washing procedure.

BACKGROUND: To determine the presence of human immunodeficiency virus-1 (HIV-1) viral RNA/DNA in whole semen, in properly isolated seminal fractions and in spermatozoa after swim-up, by extractive nested PCR and to compare the detection of HIV DNA by in situ PCR (IS-PCR) with the results of nested PCR. METHODS: We tested HIV-1 RNA and DNA by nested PCR in semen and in seminal fractions from 55 patients. Non-spermatic cells and spermatozoa pellet fractions from 10 HIV-1-positive and five HIV-1-negative men were tested for proviral DNA by IS-PCR. RESULTS: All samples of spermatozoa recovered after sperm washing were free of HIV RNA. HIV RNA tested positive in seven (13%) seminal plasma samples and only in two (4.2%) whole semen of these same samples. Of the seven seminal plasma samples testing positive for HIV RNA, four men had elevated blood viral load and three an undetectable viraemia. HIV DNA by IS-PCR turned positive in three of five samples in semen of HIV-noninfected men. CONCLUSION: HIV RNA/DNA detection in the semen of HIV-infected men proves the efficacy of sperm washing with swim-up of spermatozoa. It is recommended that nested PCR be conducted on purified seminal compartments. IS-PCR is inadequate for detecting HIV in semen.

Adult↗

PubMed1999

Are cannabinoids detected in hair after washing with Cannabio shampoo?

Today, cannabis plants are used in shampoo preparations, in foodstuffs (e.g., oils, noodles, crackers, etc.), and in beverages (e.g., tea). These products often contain < 1% delta9-tetrahydrocannabinol (THC) in order to eliminate psychoactive effects, but some of them can include 1 to 3% of THC. Gas chromatography-mass spectrometry (GC-MS) analysis of Cannabio shampoo revealed the presence of THC (412 ng/mL) and two constituents of cannabis plants, cannabidiol (CBD, 4079 ng/mL) and cannabinol (CBN, 380 ng/mL). In order to verify if normal hygiene practices with Cannabio shampoo can result in positive tests for cannabinoids in hair, three subjects washed their hair with this shampoo once daily for two weeks. After this period, hair specimens were collected. In the three hair specimens, THC, CBD, and CBN were never detected within their limits of detection, 0.05, 0.02, and 0.01 ng/mg, respectively. We concluded that the use of Cannabio shampoo during normal hygiene practices cannot be considered as a source of potential contamination of hair. In a second experiment, drug-free hair specimens (200 mg) were incubated in 10 mL water/Cannabio shampoo (20:1, v/v) for 30 min, 2 h, and 5 h. After incubation, hair strands were washed with water and separated into two portions. One portion was extracted directly; the second was decontaminated with methylene chloride and then extracted. After an incubation period of 30 min, the analysis of hair by GC-MS did not reveal the presence of THC, CBD, and CBN in hair, regardless of whether the hair was decontaminated. After an incubation period of 2 h, specimens tested positive for CBD (0.11 ng/mg without decontamination and 0.10 ng/mg with decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). After an incubation period of 5 h, specimens tested positive for CBD (0.25 ng/mg without decontamination and 0.14 ng/mg after decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). In all cases, THC was never detected. Extensive but unrealistic use of Cannabio shampoo can cause drug-free hair to test positive for CBD and CBN but not for the primary psychoactive drug THC.

Animals↗

PubMed1991

Evaluation of intrauterine insemination with washed spermatozoa from the husband in the treatment of infertility.

In this study, we examined the efficacy of intrauterine insemination with washed spermatozoa from the husband (AIH/IUI) in the treatment of infertility. A total of 127 treatment cycles were completed (1.95 cycles per patient). The indications for AIH/IUI were male subfertility (group I: 53 couples), cervical factor (group II: four couples), male and cervical factor (group III: six couples) and unexplained infertility (group IV: two couples). Sperm washing caused a reduction of the sperm concentration from 52 x 10(6)/ml to 44 x 10(6)/ml (P less than 0.08) and motility from 49% to 45% (P less than 0.03). Twelve pregnancies were achieved (18.5% per couple and 9.5% per cycle): eight pregnancies in group I (15.1%), one in group II (25%) and three in group III (50%). Of the 12 pregnancies, eight occurred in the first insemination cycle, three in the second and one in the third (1.4 cycles/pregnancy). Sperm motility was significantly higher in the pregnant than in the non-pregnant group (65.5% versus 46.4%, respectively; P less than 0.004), whereas no significant difference was observed in the sperm concentration between these two groups (39 x 10(6)/ml and 54 x 10(6)/ml, respectively). In conclusion, AIH/IUI is a technically easy and non-invasive procedure which offers a satisfactory pregnancy rate in selected couples with male subfertility and/or cervical factor infertility.

Adult↗

PubMed1993

The techniques and complications of out-patient uterine washing in the assessment of endometrial function.

The complications of a consecutive series of 90 uterine washings performed among 52 women were studied. The amount of discomfort was significantly less than that of endometrial biopsy but greater than that of cervical smear or blood sampling. Incomplete recovery of fluid (< 8 ml from 10 ml flushing fluid) was reported in only 3% of cases. Vaginal bleeding, pelvic infection and delayed abdominal pain were not encountered. The techniques of uterine washing are simple and may have important implications in the study of endometrial function.

Adult↗

PubMed1986

The pulse wash instrument. A new sampling method for uterine cervical cancer detection.

A new sampling method for uterine cervical cancer detection is described. In this method, sampling of cytologic material is done by using a pulse wash instrument. Liquid jets with a diameter of 0.2 mm at a speed of 20 m/s create a successful rinsing effect of cervical epithelial cells due to the high kinetic energy produced. Because cells are suspended in the flushing liquid it is possible to collect material for additional cytochemical, immunocytochemical, and microbiologic diagnostic techniques in addition to a conventional smear technique. Compared to a conventional Papanicolaou smear technique performed in 75 women at two cervical atypia clinics at the Karolinska Hospital, the pulse wash technique is suggested to result in a more representative cellular sample, thus offering a method to decrease false negative diagnoses in uterine cervical cancer detection.

Adolescent↗

PubMed1999

Four deaths due to carbon monoxide poisoning in car washes.

In a period of 13 months, three separate incidents of lethal carbon monoxide (CO) poisoning in closed car wash bays resulted in the deaths of 4 white men aged 20 to 36 years. Each man appears to have been intoxicated with mind-altering substances, which may impair judgment, perception of outside conditions, and self-awareness. All four died in winter months. For three men, the deaths were ruled accidental, and for the remaining man, the previous deaths appear to have provided a model for suicide. Warning signs may not be effective to prevent future CO deaths in car washes because of the possible role of intoxication. Mechanical or electronic methods to prevent a bay door from closing completely may be preferable.

Adult↗

PubMed1990

A comparison of washed red blood cells versus packed red blood cells (AS-1) for cardiopulmonary bypass prime and their effects on blood glucose concentration in children.

The effects on blood glucose concentrations of packed red blood cells (AS-1) (group I) versus washed red blood cells (group II) for cardiopulmonary bypass prime were compared in 20 infants weighing less than 10 kg undergoing cardiac surgical procedures. All patients were anesthetized with N2O/O2/isoflurane/fentanyl and received lactated Ringer's solution prior to bypass. Blood glucose concentrations prior to bypass were 85 +/- 15 mg/dl (mean +/- SD) in group I and 81 +/- 14 mg/dl in group II. Blood glucose concentrations were 210 +/- 21 mg/dl versus 78 +/- 14 mg/dl (P less than 0.001) 10 min after initiation of bypass, 241 +/- 48 mg/dl versus 107 +/- 28 mg/dl (P less than 0.001) prior to separation from bypass, and 214 +/- 52 mg/dl versus 97 +/- 19 mg/dl (P less than 0.001) after protamine administration in group I and group II, respectively. The use of washed red blood cells for cardiopulmonary bypass priming solution in infants significantly attenuates the increase in blood glucose concentration otherwise observed during cardiopulmonary bypass.

Adenine↗

PubMed1999

Evaluation of sperm washing as a potential method of reducing HIV transmission in HIV-discordant couples wishing to have children.

OBJECTIVE: A number of discordant couples, in whom the man is HIV positive and the woman is HIV negative, wish to have children. To conceive they must abandon protected sex, posing a risk of HIV transmission to the woman and so to the child. In such circumstances purification of spermatozoa ('sperm-washing') to inseminate the woman artificially has been proposed as a method of reducing the risk of transmission. Here we evaluate whether this does represent a true risk reduction. METHODS: Semen samples from HIV-positive patients were separated into spermatozoa, non-sperm cells (NSCs) and plasma fractions. The amount of viral RNA present in each fraction was measured and compared with the level in the peripheral blood. Each fraction was also assessed for the presence of proviral DNA. The ability of spermatozoa to be infected was assessed by evaluating for the presence of HIV receptors, i.e. CD4, CCR5 and CXCR4 on the surface of the sperm, by flow cytometry. RESULTS: A poor correlation was found between the levels of HIV in blood and semen. Within the semen the virus was restricted to the plasma and/or NSCs. All spermatozoa were negative for viral RNA or proviral DNA. Spermatozoa did not express significant levels of CD4, CCR5 or CXCR4, suggesting that they are unlikely to be major targets for HIV infection. CONCLUSIONS: These data suggest that spermatozoa are not major targets of HIV infection. Purifying spermatozoa reduced the level of HIV RNA and proviral DNA to below the detection limit of the assays irrespective of the amount of virus present in the unfractionated semen. On the basis of these data we would recommend 'sperm-washing' followed by insemination as a safer alternative to natural conception for HIV-discordant couples wishing to have children.

DNA, Viral↗

PubMed1996

Model testing using Chernobyl data: I. Wash-off of 90Sr and 137Cs from two experimental plots established in the vicinity of the Chernobyl reactor.

The "Wash-off" scenario is designed to test models concerned with the movement of trace contaminants from terrestrial sources to bodies of water, specifically the contamination of surface water by wash-off of radionuclides initially deposited onto soils. Particular emphasis is placed on chemical speciation and on the geochemical and geophysical processes affecting transfer of contaminants from soil to water. The scenario gives descriptions of two experimental plots near the Chernobyl power plant, one using heavy rain and one using snow melt, together with characteristics of the initial aerial deposition of the radionuclides and data on topography, soil type and characteristics, and time-varying precipitation. Predictions are requested for (1) the vertical distribution of concentrations of exchangeable and nonexchangeable forms of 137Cs and 90Sr in the soil of the experimental plots, (2) concentrations of 137Cs and 90Sr in runoff water from the experimental plots, and (3) total amounts of 137Cs and 90Sr removed by runoff from the experimental plots. Test data (field measurements) are available for all endpoints.

Cesium Radioisotopes↗

PubMed1987

Endosalpingiosis in female peritoneal washings: a diagnostic pitfall.

Endosalpingiosis in female peritoneal washings can be a source of false-positive diagnoses, especially in patients with known ovarian carcinoma. In this article we report the features of endosalpingiosis in the peritoneal washings of four patients, three of whom were initially reported as "suspicious for malignancy." Two of the patients had ovarian serous tumor of borderline malignancy; however, the tumors were confined to the ovaries with no evidence of disseminated peritoneal malignancy. All four patients had histologic evidence of pelvic endosalpingiosis. It is important to recognize this entity cytologically to avoid a misdiagnosis of recurrent or disseminated malignancy.

Adult↗

PubMed1986

Wringer washing machine injuries: criteria for obtaining radiological studies.

While the number of wringer washing machine injuries is declining due to the increasing use of automatic washing machines, these injuries still occur. Historically, children with wringer injuries received routine radiographs to diagnose fractures, in spite of the low fracture rate in wringer injuries. Review of the records at Milwaukee Children's Hospital between the years 1973 and 1983 revealed that of the 99 wringer injuries seen, 80 of 99 patients were radiographed and only five fractures were diagnosed. Of these fractures only two were attributable to the wringer device and these two required therapy. Criteria are developed to identify which patients should undergo radiologic studies. Using the criteria of localized pain or tenderness, crepitation, obvious deformity, or abnormal movement in the extremity, the two serious fractures would have been identified, and these two patients would have been radiographed. The other 78 patients would not have been radiographed.

Accidents, Home↗

PubMed1986

Flow cytometry of peritoneal washings in gynecologic neoplasia.

A prospective, double blind study was initiated to compare flow cytometry to cytopathology in detecting malignant cells within peritoneal washings. Deoxyribonucleic acid (DNA) histograms were generated using 4',6-diamidino-2-phenylindole (DAPI) as a DNA fluorochrome. Evaluation of these data revealed a correlation of 84.5% with the cytologic findings. Two specimens demonstrated euploidy in the presence of cytologically malignant cells (false negative), 28 of 128 specimens (21.8%) manifested aneuploidy with negative cytologic findings (false positive). Further evaluation of this latter subgroup revealed 22 of the 28 to possess unequivocal histologic evidence of malignancy, thus yielding an actual false positive rate of 4.7%. This preliminary study demonstrates that flow cytometry is a highly sensitive, accurate, and analytic method for the detection of malignant cells within peritoneal washings and that it may augment the cytologic examination. However, additional comparative studies are necessary to conclusively demonstrate its apparent diagnostic potential.

Cell Nucleus↗

PubMed2000

Nasal wash technique for nasal foreign body removal.

Nasal foreign bodies are seen commonly both in the office and pediatric emergency department setting. There have been a number of strategies described for their removal. We describe the "nasal wash" as a technique for nasal foreign body removal in the following three case reports. The "nasal wash" has been used in many pediatric vaccine studies as a method to collect mucus and relies on simple equipment readily available in the office setting.

Animals↗

PubMed1981

Peritoneal washing therapy for pseudomyxoma peritonei.

Two patients with pseudomyxoma peritonei (PP) had repeated intraperitoneal washing with 5% dextrose in water. Both responded to this therapy with decrease in abdominal girth. The second patient is alive, without abdominal distention, more than 112 weeks since beginning peritoneal washing therapy (PWT) for ovarian PP. Biochemical assays of urine before and after PWT show that acid mucopolysaccharide is excreted in the urine, probably after being absorbed through the peritoneal surface. The long-term effects of this procedure on renal function are not known, but PWT appears to be reasonable and safe ambulatory therapy for PP.

Adenocarcinoma↗

PubMed2003

Gastric cancer cell detection in peritoneal washing: cytology versus RT-PCR for CEA transcripts.

This study investigates the sensitivity and specificity of cytology, qualitative, and real-time RT-PCR methods in free cancer cell detection of peritoneal washing from gastric cancer patients. Peritoneal washings were collected from 65 gastric cancer patients for routine cytology and total RNA extraction for qualitative and real-time RT-PCR for CEA. The sensitivity and false-positive rate was 51.1%, 0% for cytology, 48.9% and 5% for qualitative RT-PCR for CEA, and 42.5% and 5% for real-time RT-PCR for CEA. The qualitative and real time RT-PCR results show high concordance rate (89.7%). The highest sensitivity was obtained by the combination of cytology with qualitative RT-PCR for CEA (70.2%). RT-PCR results were positive in 63.6% of cytologic "atypia" cases. Combination of cytology and either of the RT-PCR methods resulted in significantly higher sensitivity than any one of the three methods alone (P < 0.05). There was no definite advantage of the real-time RT-PCR over the conventional RT-PCR.

Adenocarcinoma↗