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Effects of cold storage on detection of avian infectious bronchitis virus in chicken carcasses and local antibodies in tracheal washes.

In order to test the survivability of infectious bronchitis virus (IBV) in dead chicken carcasses during 24 h of cold storage, 7 week-old specific-pathogen-free chickens were infected with virulent IBV Massachusetts strain M41, and were killed humanely 10 days later. Carcasses were stored in a cold room at 4 degrees C. After 1, 3, 6, 9, 12 or 24 h of storage, necropsies were carried out. Trachea, lung, kidney and rectum were collected for virus isolation by tracheal organ culture (TOC) or embryonated chicken eggs (ECE), and detection by nested reverse-transcriptase polymerase chain reaction (RT-PCR). IBV was detected by RT-PCR at all sampling times, except for 1 and 6 h of storage in kidney and 9 h of storage in kidney and rectum. For ECE, isolation was obtained at all sampling points, except at 1 and 24 h of storage in lungs. Isolation by tracheal organ cultures was less successful, except from rectum. In addition to sampling for virus, tracheal washes were collected from each carcass to measure the ability to detect local antibodies after storage. Levels of IgA in tracheal washes remained high for up to 9 h of storage, suggesting that accurate sampling for research purposes when required must be carried out within this time.

Animals↗

Ethanol washing does not attenuate the hypocholesterolemic potential of soy protein.

OBJECTIVE: A feeding study in rats investigated the principal active component for the hypocholesterolemic effect of soy protein isolate (SPI) by comparing the effect before and after ethanol washing. METHODS: Five-week-old male Sprague-Dawley rats were fed cholesterol-enriched AIN-93G diets containing 20% casein (CAS), 20% SPI, 20% ethanol-washed SPI (EWS), 18.4% EWS plus 1.6% ethanol extract (EE), or 20% CAS plus 1.6% EE for 2 wk. RESULTS: Plasma cholesterol concentrations in rats fed EWS and SPI were comparable and were significantly lower than those in rats fed CAS. The addition of EE to EWS and CAS did not influence plasma cholesterol level. Fecal steroid excretion of the three SPI groups was higher than that of the two CAS groups. The addition of EE to EWS and CAS showed a tendency to increase acidic steroid and decrease neutral steroid. CONCLUSIONS: In this experiment which used identifiable protein preparations, a significant fraction of the cholesterol-lowering effect of SPI in rats was attributed to its protein component but not to the ethanol-extractable minor constituents including isoflavones.

Animals↗

Effect of ultrasonic energy in washing of medical surgery gowns.

Today, the use of medical textiles is very important for health and hygienic purposes. Although some of medical textile materials are disposable, surgery gowns are usually laundered and re-used. After the laundering it is desirable to have these gowns keeping some of their properties such as anti-microbial and tenacity properties. For this in mind, different surgery gowns were washed and compared. In this work, a special blend of polyester carbon anti-microbial and other surgery gowns were stained with blood and later washed with both conventional method and with ultrasonic energy. The results were also analysed spectrophotometrically and the tenacity of the fabrics were determined.

Carbon↗

Serum-free influenza virus production avoiding washing steps and medium exchange in large-scale microcarrier culture.

A complete serum-free process without washing steps and medium exchange before infection for influenza A virus vaccine production (equine and human) is described for cultivation in roller bottles and in a 5-L stirred tank microcarrier system. Adherent Madin-Darby canine kidney cells (MDCK) were adapted from growth in serum containing GMEM medium to growth in serum-free Ex-Cell MDCK medium. Roller bottle experiments showed that the medium exchange step, typically required for serum containing vaccine production processes, could be omitted without losses in virus titre and without limitations in glucose or glutamine supply in the cultivation medium. The serum-free medium could even be used glutamine-free as it contained pyruvate, resulting in very low levels of ammonia. Cell attachment onto microcarriers was critical. Therefore, microcarriers had to be preconditioned in medium. Also, trypsin concentration used for inoculum preparation had to be reduced. After these modifications 1.3 x 10(6)cells/mL were obtained after 97 h (2g/L Cytodex 1) of cell growth. Maximum virus titres of 2.3-2.9 log HA units/100 microL were obtained from infections with a multiplicity of infection (moi) of 0.05-0.10 for human and equine influenza A virus. Metabolite and amino acid profiles as well as on-line data for the serum-free process are compared with the serum containing process. Omission of the medium exchange before infection clearly simplified the process and reduced sterility risks due to washing steps.

Ammonia↗

Evaluation of a monoclonal antibody-based enzyme-linked immunosorbent assay for detection of Campylobacter fetus in bovine preputial washing and vaginal mucus samples.

A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was described and evaluated for use as a presumptive screening test for detection of Campylobacter fetus in bovine preputial washing and vaginal mucus samples. A total of 725 diagnostic samples collected in the field and submitted in Clark's transport enrichment medium (TEM) were analyzed. Cultural isolation of C. fetus was used as the standard for comparison. After incubation of the TEM vials for 4-5 days, fluid was removed for culture and ELISA testing. A sandwich ELISA format was used and the target antigen was C. fetus lipopolysaccharides (LPS). A rabbit anti-C. fetus polyclonal antiserum was used as the capture antibody. Murine monoclonal antibodies (MAbs) to C. fetus serotype A and B LPS core and O-polysaccharides and a goat anti-mouse horseradish peroxidase conjugate were used as detection antibodies. ELISA and culture results for the diagnostic samples were in complete agreement. Seven hundred and eight samples were negative by both tests. All 17 culture positive samples were positive by ELISA with a MAb to LPS core. The ELISA with MAbs to LPS O-polysaccharides detected all culture positive samples with the homologous C. fetus serotype. Sixty-six preputial wash samples from three known C. fetus culture positive bulls were also analyzed. Forty-nine of these samples were positive by both ELISA and culture, 16 were positive by ELISA only, and one was negative by both ELISA and culture. The results indicate that this ELISA is useful as a screening test for the detection of C. fetus in diagnostic samples.

Animals↗

Optimization of the extended terminal subfluidization wash (ETSW) filter backwashing procedure.

The increased passage of particles and microorganisms through granular media filters immediately following backwashing is a common problem known to the water treatment community as filter "ripening" or maturation. While several strategies have been developed over the years to reduce the impact of this vulnerable period of the filtration cycle on finished water quality, this research involves a recently developed filter backwashing strategy called the extended terminal subfluidization wash (ETSW). ETSW is a method of terminating the backwash cycle with a subfluidization wash for a period of time sufficient to pass one theoretical filter-volume of water upward through the filter. ETSW was shown to remove significantly greater quantities of backwash remnant particles thereby reducing the magnitude of filter ripening turbidity and particle count spikes. Optimum ETSW flow rates were determined for deep-bed anthracite and granular activated carbon filters herein by monitoring filter effluent turbidities and particle counts during the filter ripening period. Optimality of the coagulation process was also shown to influence the magnitude of filter ripening particle passage. ETSW was found to be equally effective for biological and conventional deep-bed anthracite filters.

Filtration↗

Effect of humic acid on the photolysis of the pesticide atrazine in a surfactant-aided soil-washing system in acidic condition.

The photolytic destruction of Atrazine (ATZ) following a surfactant-aided soil-washing process was investigated in the presence of humic acid (HA). A non-ionic surfactant, Brij 35, was found to be a good solving agent, extracting ATZ without causing any retardation effect on the photolysis process. However, the HA that was co-extracted from the surfactant-aided soil-washing system was found to be capable of improving the photolysis of ATZ at low concentrations, while quenching the photodegradation of ATZ at higher concentrations. By considering the light attenuation effect due to surfactant and HA, the quantum yields of the system with respect to the proposed reaction mechanisms of the associated excited states of ATZ were investigated and modeled. The relative kinetic rates of the dominant reaction mechanisms (i.e., deactivation of triplet, direct photolysis of triplet, product formation, and quenching of triplet) were then compared.

Atrazine↗

Chlorhexidine versus sterile water vaginal wash during labor to prevent peripartum infection.

OBJECTIVE: Our purpose was to determine whether a dilute solution of chlorhexidine used as a one-time vaginal wash intrapartum can reduce the incidence of intraamniotic infection or endometritis in laboring pregnant women. STUDY DESIGN: Term pregnant women in labor were prospectively randomized to receive either 20 ml of 0.4% chlorhexidine (n = 481) or 20 ml of sterile water (n = 466) placebo. All patients were monitored for risk factors associated with intraamniotic infection. Continuous variables were compared with the Mann-Whitney U test and discrete variables were compared with Fisher's exact test. RESULTS: No significant differences in infection were found between the chlorhexidine and placebo groups. During the study period 21 of 466 women (4.5%) had intraamniotic infection in the control group compared with 25 of 481 women (5.2%) receiving chlorhexidine (p = 0.65, 95% confidence interval 0.82 to 1.41). Nine women in the placebo group (1.9%) and 9 women in the chlorhexidine group (1.9%) had endometritis (p = 1.0, 95% confidence interval 0.62 to 1.56). CONCLUSIONS: Our findings suggest that a one-time 0.4% chlorhexidine vaginal wash does not decrease the incidence of infectious morbidity in parturients, compared with the use of sterile water.

Adult↗

Prognostic significance of carcinoembryonic antigen levels in peritoneal washes in patients with gastric cancer.

BACKGROUND: Peritoneal metastasis is the most frequent cause of death in patients with gastric cancer. Detection of free cancer cells in the peritoneal cavity at the time of surgery, therefore, is considered to be of great value in predicting the peritoneal recurrence and accordingly in the prognosis in patients with gastric cancer. This study examined the clinical significance of intraoperative determination of carcinoembryonic antigen (CEA) levels in peritoneal washes (pCEA) in patients with gastric cancer. METHODS: CEA levels in peritoneal washes were correlated retrospectively with several clinicopathologic factors including clinical outcome in 56 patients with resectable gastric cancer. RESULTS: Among several clinicopathologic factors, the depth of tumor invasion significantly and independently correlated with pCEA levels as revealed by multivariate stepwise logistic regression analysis. A significant difference in overall survival rates was observed between pCEA-positive and pCEA-negative groups: 5-year survival rates were 95.7% in pCEA-negative and 20% in pCEA-positive patients (P <0.0001). Multivariate analysis indicated that pCEA level is a statistically significant independent prognostic factor for the survival of patients with gastric cancer, and is an important factor for predicting peritoneal recurrence. CONCLUSIONS: pCEA could be a potential predictor of a poor prognosis as well as peritoneal recurrence in patients with gastric cancer. We believe that this information could contribute to determining the optimal intraoperative and postoperative therapeutic plan including adjuvant chemotherapy of gastric cancer.

Adult↗

Diagnosis for interstitial lung disease in patients with acquired immunodeficiency syndrome (AIDS): a prospective comparison of bronchial washing, alveolar lavage, transbronchial lung biopsy, and open-lung biopsy.

This study was undertaken to compare prospectively the diagnostic yield of the various bronchoscopic techniques with that of open-lung biopsy for interstitial lung disease in patients with acquired immunodeficiency syndrome (AIDS). Under general anesthesia, 15 patients sequentially underwent bronchial washing, transbronchial lung biopsy, alveolar lavage, and open-lung biopsy in the same segment of lung. Of nine patients with Pneumocystis carinii, seven were diagnosed by means of the transbronchial lung biopsy, eight by the open-lung biopsy, and all nine by alveolar lavage. Of the six patients with cytomegalovirus, five were diagnosed by the open-lung biopsy, five by the transbronchial lung biopsy, and three by alveolar lavage. The sensitivities of the procedures for identifying infection were washings (15%), transbronchial lung biopsy (50%), alveolar lavage (73%), and open-lung biopsy (88%). Combined, transbronchial lung biopsy and alveolar lavage showed a diagnostic yield (85%) for infections comparable to that of open-lung biopsy (88%), thereby obviating the need for open-lung biopsy for such diagnoses. However, open-lung biopsy was the only procedure that diagnosed Kaposi's sarcoma in lung.

Acquired Immunodeficiency Syndrome↗

The use of saline-washed frozen red cells for extracorporeal bypass.

The use of saline-washed frozen red cells and homologous whole blood during cardiopulmonary bypass was compared in two groups of patients with similar medical disease, sex, age, pump time, types of operation, and surgical techniques. A technique for using saline-washed frozen red cells for cardiopulmonary bypass with no increase in hemolysis is described. Increased platelet counts were present at the end of bypass in the patients who were given frozen cells. The availability and the many advantages of frozen cells, such as improved oxygen transport and the absence of homologous blood reactions and viral infections, make them the preferred cellular blood component for use during open-heart operations.

Blood Cell Count↗

The affinities of prostaglandin H2 and thromboxane A2 for their receptor are similar in washed human platelets.

Both thromboxane A2 (TXA2) and its precursor prostaglandin H2 (PGH2) are labile and share a common receptor. The affinities of these two compounds for their putative common receptor are unknown. We compared the potencies of TXA2 and PGH2 to aggregate human platelets and bind to the TXA2/PGH2 receptor. TXA2 was more potent than PGH2 in initiating aggregation in platelet-rich plasma, EC50 of 66 +/- 15 nM and 2.5 +/- 1.3 microM, respectively. In washed platelets, however, PGH2 was more potent than TXA2 with EC50 values of 45 +/- 2 nM and 163 +/- 21 nM, respectively. The affinity of these two compounds in washed platelets was determined in radioligand competition binding assays employing [125I]-PTA-OH. The Kd values for PGH2 and TXA2 were 43 nM and 125 nM, respectively. The results demonstrate that the affinity of PGH2 for the platelet TXA2/PGH2 receptor is greater than previously thought. The data raise the possibility that PGH2 may significantly contribute to the responses attributed to TXA2 in vivo.

Binding, Competitive↗

Variation of movement characteristics with washing and capacitation of spermatozoa. II. Multivariate statistical analysis and prediction of sperm penetrating ability.

Fifty semen samples were studied by computer-assisted semen analysis before testing in the sperm penetration assay (SPA). Twenty-one concentration and movement measurements were obtained from each sample on sperm in semen, after washing and swim-up, and again after an 18-hour capacitation period. Discriminant analysis was then used to define a function from these measurements that would classify the SPA results as above or below a 10% penetration rate. A significant function was identified using the following variables: sperm concentration and motility in semen, and mean curvilinear velocity, linearity, and amplitude of lateral head displacement of washed sperm. Movement measurements of capacitated spermatozoa were not useful predictors in this analysis. The overall accuracy of this function for predicting SPA results was 72%. These findings demonstrate that computer-derived measurements of sperm movement provide biologically useful information regarding sperm function, and, in addition, emphasize the importance of multivariate techniques in the analysis and description of human sperm motion.

Analysis of Variance↗

Treatment of refractory infertility by transcervical intrauterine insemination of washed spermatozoa.

One hundred thirteen couples with either male factor, cervical factor, endometriosis, or idiopathic infertility of at least 3 years' duration were treated by intrauterine insemination (IUI) of washed motile sperm. Of the 68 women who became pregnant or completed at least three cycles of insemination, the overall pregnancy rate was 38.2%, with a mean of 1.7 treatment cycles per pregnancy. The average pregnancy rate per treatment cycle was 11.4%. Women who did not become pregnant underwent an average of 4.7 treatment cycles. Importantly, only two pregnancies occurred independent of treatment in 113 couples. In the male factor group, the pregnancy rate was 42.9% (n = 21). In women with a cervical factor, 34.5% became pregnant (n = 29); in idiopathic infertile couples or women suffering from endometriosis, there was a pregnancy rate of 38.9% (n = 18). The presence of sperm antibodies in either the male or female partner significantly lowered the pregnancy rate (6.7%) when compared with couples without sperm antibodies (50.0%). The authors conclude from these observations that IUI with washed sperm is a successful mode of therapy, especially in the case of males with asthenozoospermia.

Female↗

Artificial insemination: a comparison of pregnancy rates with intrauterine versus cervical insemination and washed sperm versus serum swim-up sperm preparations.

A four-cycle artificial insemination protocol was undertaken, with luteinizing hormone (LH) timing, to compare washed sperm and serum swim-up sperm preparations and cervical and intrauterine insemination. Of the 75 patients entered into the protocol series, 63 finished all 4 cycles. During the treatment period, 24 patients became pregnant, 20 of whom were donor (AID) pregnancies (out of 52 patients) and 4 were husband (AIH) pregnancies (out of 11 patients). Seventy-five percent of the pregnancies were intrauterine, whereas only 25% were from cervical inseminations. Sixty-two percent of the pregnancies were due to serum swim-up sperm preparations, and 38% were due to washed sperm preparations. The data indicate substantially higher pregnancy rates can be obtained with intrauterine insemination and suggest that additional studies are necessary to determine if the method of sperm preparation significantly influences pregnancy rates.

Cervix Uteri↗

Variation of movement characteristics with washing and capacitation of spermatozoa. I. Univariate statistical analysis and detection of sperm hyperactivation.

Movement alterations as spermatozoa are washed and capacitated in preparation for the zona-free sperm penetration assay were investigated using a videomicrographic computer-assisted system for tracking and analysis of sperm trajectories. Thirty-three semen samples from infertile men and 17 fertile controls were studied. Despite significant differences on the sperm penetration assay, univariate comparisons of movement characteristics between infertile and control samples failed to detect differences. For all samples combined, significant alterations in movement characteristics were detected after washing. After an 18-hour sperm capacitation, mean velocity, head yawing amplitude, and head yawing frequency still were significantly increased over seminal plasma values. Videomicrographic motion analysis thus allows detection of changes that are consistent with hyperactivated motility, as has been described for capacitating spermatozoa.

Diagnosis, Computer-Assisted↗

Hypocellular bladder wash specimens and their clinical significance.

Bladder wash flow cytometry specimens containing less than 1,000 cells were defined as hypocellular. Between June 1981 and December 1990, 5,676 bladder wash flow cytometry specimens were collected, of which 142 (2.5%) were deemed hypocellular. Of the 132 evaluable patients with hypocellular specimens 34 (30%) had documented evidence of carcinoma compared to 52% for the entire population. Among the 34 patients 30 had transitional cell carcinoma, of which 12 were carcinoma in situ, 9 stage Ta or T1 and 9 stage T2 or greater. Thus, there was no obvious relationship between stage or grade of carcinoma versus probability of a hypocellular specimen. Differences in irrigation techniques of different urologists in the operating room using general anesthesia versus catheter and cystoscopic irrigation in the outpatient clinic are discussed as possible explanations for the hypocellular specimens.

Biopsy↗

A new technique for bladder washing.

We describe a simple adaptation of the Water Pik (Teledyne Water Pik, Fort Collins, Colorado) irrigating device which allows vigorous, direct-vision agitation of the bladder wall. Three groups of mongrel dogs were subjected to cystoscopy and either syringe barbotage, half-speed Water Pik irrigation, or full-speed Water Pik irrigation of the bladder wall. Transitional cell counts were then done on centrifuged aliquots of each bladder wash specimen. The average number of transitional cells per high-power field were similar between the control group and the syringe barbotage group (2.5 and 1.5 respectively). However, both the half-speed and the full-speed Water Pik groups demonstrated statistically higher cell counts (5.7 and 13.7) when compared to both the controls and syringe barbotage groups. We conclude that Water Pik irrigation is an effective method to increase cell yield in bladder wash specimens.

Animals↗