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Studies on leakage of enzymes from washed bull and ram spermatozoa.

Washing of bull and ram spermatozoa resulted in significant losses of lactic dehydrogenase (LDH) and glucose phosphate isomerase (GPI) from the cell suspensions. Re-suspension of washed bull spermatozoa caused an immediate release of enzymes from the cells. Preincubation of washed ram spermatozoa with 0.025% formaldehyde increased GPI levels but decreased LDH concentration in the extracellular fluid while hexokinase release was unaffected. Varying the incubation temperature between 20 and 37 degrees C affected extracellular LDH and GPI levels. It is suggested that enzyme release from spermatozoa may occur in the absence of any apparent cellular damage.

Animals↗

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein.

The dye 1-anilino-naphthalene-8-sulphonate (ANS) bound at 94.5 +/- 2.7 pmol per 10(6) spermatozoa (KD = 1.83 +/- 0.13 X 10(-5) M) and 7.30 +/- 0.17 pmol per mg seminal plasma protein (KD = 2.19 +/- 0.06 X 10(-5) M). Equilibration of excess ANS with a mixture of washed spermatozoa and seminal plasma resulted in a significant increase in fluorescence over that calculated by summation of the individual levels (P less than 0.05). A less pronounced but significant increase was seen when an egg-yolk lipoprotein, previously shown to have cryoprotective properties, was similarly added to washed spermatozoa (P less than 0.05). This increase in fluorescence was not reduced by washing spermatozoa--lipoprotein mixtures, suggesting that an interaction between the lipoprotein and the cell membrane had occurred.

Anilino Naphthalenesulfonates↗

Concentrations of steroid hormones, and of prolactin, in washings of the human uterus during the menstrual cycle.

Levels of steroid hormones, prolactin and protein were determined in trans-cervical flushings of uteri of 73 consenting women presenting for reversal of sterilization. Median total levels of steroids (pmol), prolactin (mu i.u.) and protein (mg) in the washings were: pregnenolone, 4.22; pregnenolone sulphate, 15.1; progesterone, 1.01; dehydroepiandrosterone (DHEA), 8.92; DHEA sulphate, 368; androstenedione, 2.23; testosterone, 1.04; oestrone, less than 0.7; oestrone sulphate, 0.49; oestradiol, 0.08; prolactin, 23.8; and protein, 5.75. Levels of these components of uterine flushings did not vary significantly between Days 6-10, 11-14, 15-20 and 21-28 after the onset of the previous menstrual period (P greater than 0.05). Uniform levels of free steroids in uterine washings throughout the menstrual cycle, and low free steroid/total protein ratios (all less than 3 pmol/mg), support other evidence for a paucity of steroid-binding proteins in human histotroph. The predominance of DHEA sulphate and of pregnenolone sulphate in human uterine washings is in accord with their abundance in plasma, and may provide an important precursor pool for de-novo steroidogenesis by human embryos before implantation. Our results support the view that human histotroph is a filtrate of plasma.

Dehydroepiandrosterone↗

Clinical subtypes of obsessive-compulsive disorder based on the presence of checking and washing compulsions.

OBJECTIVE: We aimed at examining the utility of checking and washing compulsions as markers of valid subtypes of obsessive-compulsive disorder (OCD). METHODS: One hundred and six patients with obsessive-compulsive disorder were evaluated with a socio-demographic and clinical questionnaire, the Structured Clinical Interview for DSM-IV, the Yale-Brown Obsessive-Compulsive Scale, the Clinical Global Impression, the Beck Depression Inventory, the Hamilton Rating Scale for Depression, and the Global Assessment of Functioning. These individuals were allocated in one of four subgroups [checkers (OCD-Ch; n = 20), washers (OCD-Wa; n = 13), checkers and washers (OCD-CW; n = 48), and non-checkers and non-washers (OCD non-CW = 25)] on the basis of the presence and the clinical relevance of checking and/or washing compulsive behaviors across their lifetime. Socio-demographic and clinical variables were compared and contrasted between the groups by means of ANOVA followed by post-hoc Least Significant Difference or Dunnett's tests for continuous variables and chi-square tests followed by partitioned chi-square tests for categorical variables. RESULTS: OCD-Ch and OCD-Wa did not differ on most demographic and clinical features, the only exception being the number of different types of obsessions, which were significantly higher in the former group. The OCD-CW group was more likely to exhibit an insidious onset of obsessive-compulsive symptoms, to manifest itself as a mixed subtype of obsessive-compulsive disorder and to display obsessions with contamination themes. On the other hand, the OCD non-CW group was more likely to exhibit an acute onset, a shorter duration of illness, obsessions with religious themes, an episodic course, and less severe obsessive-compulsive symptoms. CONCLUSIONS: In our sample, the probing of the presence of checking and/or washing compulsions has provided significant empirical support to establish valid subtypes of obsessive-compulsive disorder.

Adolescent↗

Techniques to improve the shear strength of impacted bone graft: the effect of particle size and washing of the graft.

BACKGROUND: When fresh morselized graft is compacted, as in impaction bone-grafting for revision hip surgery, fat and marrow fluid is either exuded or trapped in the voids between particles. We hypothesized that the presence of incompressible fluid damps and resists compressive forces during impaction and prevents the graft particles from moving into a closer formation, thus reducing the graft strength. In addition, viscous fluid such as fat may act as an interparticle lubricant, thus reducing the interlocking of the particles. METHODS: We performed mechanical shear testing in the laboratory with use of fresh-frozen human femoral-head allografts that had been passed through different orthopaedic bone mills to produce graft of differing particle-size distributions (grading). RESULTS: After compaction of fresh graft, fat and marrow fluid continued to escape on application of normal loads. Washed graft, however, had little lubricating fluid and better contact between the particles, increasing the shear resistance. On mechanical testing, washed graft was significantly (p < 0.001) more resistant to shearing forces than fresh graft was. This feature was consistent for different bone mills that produced graft of different particle-size distributions and shear strengths. CONCLUSIONS: Removal of fat and marrow fluid from milled human allograft by washing the graft allows the production of stronger compacted graft that is more resistant to shear, which is the usual mode of failure. Further research into the optimum grading of particle sizes from bone mills is required.

Bone Transplantation↗

A washing procedure to mobilize mixed contaminants from soil: I. Polychlorinated biphenyl compounds.

We conducted a laboratory study to assess the feasibility of a washing process with nonionic and anionic surfactants in combination with ethylenediaminetetraacetate (EDTA) for the simultaneous mobilization of heavy metals and polychlorinated biphenyls (PCBs) from a field-contaminated soil. Unit processes consisting of complexometric extraction and surfactant-assisted mobilization were combined with reagent regeneration and detoxification steps to generate innocuous products. Ten minutes of ultrasonic mixing of the soil with a combination of 30 mL L(-1) surfactant suspension and a sparing quantity (2 mmol) of EDTA mobilized appreciable quantities of PCBs, virtually all of the available Cd, Cu, Mn, and Pb, and lesser amounts of the Zn, Ni, and Cr but only small quantities of Al and Fe. Relative to individual reagents, combinations of surfactant (Brij 98, Triton X-301, or Triton XQS-20) with EDTA did not influence PCB extraction efficiencies perceptibly. Of the three surfactants, the Brij 98 proved to be the most efficient for three successive extractions with a single charge, mobilizing 83% of the PCBs, whereas companion extractions that used fresh reagent each time mobilized 87% of the soil PCB content. The decreased PCB mobilization with the same quantity of anionic surfactant (71 or 68%) resulted from losses during the EDTA regeneration process with zero-valent Mg. In toto, these studies demonstrate that PCB compounds and selected heavy metals can be coextracted efficiently from soil with three successive washes with the same washing suspension containing EDTA and a nonionic surfactant.

Chelating Agents↗

Wash-in methodology and modeling to determine hepatocellular D-glucose transport in the perfused rat liver.

Wash-in experiments, which may be useful for the study of the disposition of substrates in the liver, have not been well described. To investigate physiological models for wash-in curves, we performed experiments on the perfused livers of male Wistar rats anesthetized with pentobarbital. Test perfusate contained (14)C-sucrose as the extracellular marker and (3)H-glucose. Liver perfusions were performed with background glucose concentrations of 5.7, 10.7, 51.2, or 108.5 mM. Outflow time-activity curves were analyzed with the use of four models. The V(max) and K(m) for the influx of glucose were 1.1 +/- 0.03 micromol/s/g and 41 +/- 3 mM with the Crone-Renkin early extraction model; 1.4 +/- 0.04 micromol/s/g and 36 +/- 3 mM with dispersion model analysis; 1.8 +/- 0.1 micromol/s/g and 25 +/- 4 mM with the Goresky distributed model to fit differentiated wash-in curves; and 2.7 +/- 0.6 micromol/s/g and 28 +/- 21 mM with compartmental analysis. There was reasonable agreement between the four models, and they yielded results similar to those reported for glucose uptake in other preparations.

Animals↗

Treatment of distillery spent-wash by ozonation and biodegradation: significance of pH reduction and inorganic carbon removal before ozonation.

This study is aimed at exploring strategies for mineralization of refractory compounds in distillery effluent by anaerobic biodegradation/ozonation/aerobic biodegradation. Treatment of distillery spent-wash used in this research by anaerobic-aerobic biodegradation resulted in overall COD removal of 70.8%. Ozonation of the anaerobically treated distillery spent-wash was carried out as-is (phase I experiments) and after pH reduction and removal of inorganic carbon (phase II experiments). Introduction of the ozonation step resulted in an increase in overall chemical oxygen demand (COD) removal, with the highest COD removals of greater than 95% obtained when an ozone dose of approximately 5.3 mg ozone absorbed/mg initial total organic carbon was used. The COD removal during phase II experiments was slightly superior compared with phase I experiments at similar ozone doses. Moreover, efficiency of ozone absorption from the gas phase into distillery spent-wash aliquots was considerably enhanced during phase II experiments.

Anaerobiosis↗

Systemic absorption of a leucovorin mouth wash: a pharmacologic study.

Mucositis is one of the major problems encountered after the administration of systemic chemotherapy. Leucovorin, routinely used as a rescue agent for methotrexate may reduce toxicity, but may also reduce the effectiveness of the chemotherapeutic agent. If leucovorin is administered as a mouth wash, local toxicity may be reduced without loss of methotrexate efficacy. In order to study this, 15 normal human volunteers were given leucovorin mouth wash and then had plasma determinations of 5-methyl-tetrahydrofolate and citrovorum factor. Small but statistically significant increases in plasma levels of 5-methyl-tetrahydrofolate were observed with no increase in levels of plasma citrovorum factor. It is concluded therefore that a small amount of leucovorin is absorbed systemically when administered as a mouth wash, but such an amount would most likely not be significant enough to reduce the effect of methotrexate therapy, but may reduce mucositis.

Absorption↗

[Effects of temperature, humidity, minor injury and washing on penetration of dermatophytes into human stratum corneum].

We evaluated the minimum time for penetration of Trichophyton mentagrophytes into human stratum corneum using an experimental model of tinea pedis. After fungal elements were applied on the surface of stratum corneum obtained from a healthy human heel, samples were incubated under designated conditions of temperature and humidity. The penetration of fungal elements was much faster at 35 degrees C than 27 degrees C despite the fact that the latter is an optimal temperature for fungal growth. At 35 degrees C and 100% humidity the minimum time required for penetration was one day. When we applied fungal elements on an abraded surface of stratum corneum, fungi penetrated within a half day under the same conditions. This suggests that minor injury of stratum corneum is a significant factor for infection. The development of tinea pedis does not occur frequently in daily life. We examined the effect of washing the surface of stratum corneum to which T. mentagrophytes had been applied. The samples were incubated under conditions simulating daily life: i.e. with 80% humidity for 8 hours, and 100% humidity for 16 hours. After washing, nearly all the fungal elements had been removed from the surface of stratum corneum within one day. The data suggests that to prevent tinea pedis, daily washing of soles and interdigital regions is effective.

Arthrodermataceae↗

Rapid detection of Salmonella in chicken washes by immunomagnetic separation and flow cytometry.

Use of flow cytometry to rapidly detect Salmonella in chicken carcass washes was investigated. A direct immunomagnetic separation method was used to prepare samples and was found to be an effective method for separating target cells from debris in chicken carcass washes. When flow cytometry was combined with immunomagnetic separation, the average lowest detectable level of Salmonella detected was 2.3 x 10(4) CFU/ml. Fifty of 100 wash samples from six groups were inoculated with 2 x 10(-1) CFU of Salmonella Typhimurium per milliliter. After 18 h of enrichment at 37 degrees C, all samples were tested for Salmonella using flow cytometry and conventional culture methods. An identification correlation of 96% was found between flow cytometry and xylose-lysine-tergitol agar plating. Quantitative analysis established a significant linear relationship between the enumeration results of flow cytometry and xylose-lysine-tergitol agar plate counts (R2 = 0.796). Time required for flow cytometry, including sample processing and flow cytometric analysis, was less than 1 h.

Animals↗

Influence of traditional brine washing of smear Taleggio cheese on the surface spreading of Listeria innocua.

The influence of a traditional procedure of washing of smear Taleggio cheese on surface spreading of Listeria innocua was studied. This practice is carried out during ripening to remove molds, to select the surface microflora, and to control the ripening process. One cheese, both of 2 (i) and 4 (ii) weeks of ripening, was surface-inoculated with approximately 3 log CFU of L. innocua per entire cheese surface. The inoculated cheeses and others of the same age were weekly washed with brine solution. Listeria was spread both on the surface of the inoculated cheese and on the other cheeses, and it was also found in the brines and on the wooden boxes where the cheeses were ripened. The time of ripening when contamination occurs influenced the behavior of Listeria. At the moment of contamination, the smear surface microflora of (i) cheese was approximately 2 log CFU/g higher than of (ii) cheese. Listeria inoculated on 2-week-ripened cheese was able to colonize the entire surface of the cheese and to cross-contaminate the other cheeses. On the contrary, Listeria inoculated on a 4-week-ripened cheese was partially spread on the surface of the originally inoculated cheese, and the transfer of contamination by the washing procedure was restrained. Because a random distribution of Listeria on cheese surface was observed, the importance of the mode of sampling was discussed. Because of the lack of critical control points during ripening of Taleggio cheese, the Listeria hazard needs to be controlled by taking appropriate control measures to break off the contamination cycle (cheese --> brine --> wooden boxes --> cheese).

Cheese↗

Efficacy and reproducibility of a produce wash in killing Salmonella on the surface of tomatoes assessed with a proposed standard method for produce sanitizers.

The reproducibility of a method developed to evaluate point-of-use sanitizers for fresh produce was tested at three different laboratories. Mixtures of five Salmonella serotypes were inoculated on the surface of ripe tomatoes. After the inoculum was dry, tomatoes were placed inside a plastic bag and sprayed with sterile USP water, Dey and Engley (D/E) neutralizer broth, or a prototype Fit produce wash (PW), an alkaline solution comprised of generally recognized as safe ingredients (water, oleic acid, glycerol, ethanol, potassium hydroxide, sodium bicarbonate, citric acid, and distilled grapefruit oil), and rubbed for 30 s. The tomatoes were rinsed 10 s with 195 ml of D/E neutralizer broth (rinse solution), then combined with 20 ml of D/E neutralizer (residual wash solution) and rubbed by hand to remove residual Salmonella. Populations of Salmonella were determined for each tomato in the rinse solution and residual wash solution. Treatment with PW resulted in reductions in the number of Salmonella 2 to 4 logs greater than those achieved with the sterile water or D/E neutralizer broth controls. Consistent results were obtained across the three study sites, indicating reproducible results were obtained using the test method. The method used to determine the efficacy of killing or removing Salmonella from tomatoes in this study is suggested as a standard method for measuring the efficacy of sanitizers on tomatoes and other similar fruits and vegetables with rigid, smooth surfaces.

Colony Count, Microbial↗

Reduction of listeria monocytogenes on green peppers (Capsicum annuum L.) by gaseous and aqueous chlorine dioxide and water washing and its growth at 7 degrees C.

Reduction of Listeria monocytogenes Scott A on uninjured and injured surfaces of green peppers after 0.3- and 3-mg/ liter gaseous and aqueous ClO2 treatment and water washing for 10 min at 20 degrees C was studied. Growth of the L. monocytogenes untreated or treated with 0.6 mg/liter ClO2 gas for 30 min at 20 degrees C on green peppers also was investigated. A membrane-surface-plating method was used for resuscitation and enumeration of L monocytogenes treated with ClO2. The bacterial viability on pepper surfaces was visualized using confocal laser scanning microscopy (CLSM). Live and dead cells of L. monocytogenes were labeled with a fluorescein isothiocyanate-labeled antibody and propidium iodide, respectively. More than 6 log CFU/5 g L. monocytogenes on uninjured surfaces and about 3.5 log CFU/5 g on injured surfaces were inactivated by both 3-mg/liter and 0.6-mg/liter ClO2 gas treatments. The 3-mg/liter aqueous ClO2 treatment achieved 3.7- and 0.4-log reductions on uninjured and injured surfaces, respectively; whereas, water washing alone showed 1.4- and 0.4-log reductions, respectively. ClO2 gas treatment was the most effective in reducing L. monocytogenes on both uninjured and injured green pepper surfaces, when compared with aqueous ClO2 treatment and water washing. The significant difference (P < 0.05) between log reductions on uninjured and injured surfaces and the results from CLSM analysis suggested that injured surfaces protected more bacteria from sanitation treatments than did uninjured surfaces. Not only could L. monocytogenes grow on green pepper surfaces at 7 degrees C, bacteria that survived the 0.6-mg/liter ClO2 gas treatment also could grow.

Capsicum↗

Survival of Salmonella in waste egg wash water.

Waste wash waters from chicken egg-processing facilities can harbor high densities of bacteria, including salmonellae. For this study, we enumerated total coliforms, Escherichia coli, and Salmonella spp. in the egg wash waters of a large egg producer. We then determined how long these organisms would survive at temperatures of 5, 15, and 25 degrees C. We found that the fraction of salmonellae surviving over time at a given temperature was comparable to the fraction of indicator organisms that survived. We also found that the survival of these organisms varied with temperature, with 16, 8, and < 2 days being required for a 90% reduction of Salmonella in waste wash water held at 5, 15, and 25 degrees C, respectively. Finally, we noted that the response of laboratory-derived cultures to environmental stresses mimics the response of the indigenous microbial population, but individual cells within that population may survive for longer periods than laboratory-cultured strains.

Animals↗

Apple quality, storage, and washing treatments affect patulin levels in apple cider.

Patulin is a mycotoxin produced primarily by Penicillium expansum, a mold responsible for rot in apples and other fruits. The growth of this fungus and the production of patulin are common in fruit that has been damaged. However, patulin can be detected in visibly sound fruit. The purpose of this project was to determine how apple quality, storage, and washing treatments affect patulin levels in apple cider. Patulin was not detected in cider pressed from fresh tree-picked apples (seven cultivars) but was found at levels of 40.2 to 374 microg/liter in cider pressed from four cultivars of fresh ground-harvested (dropped) apples. Patulin was not detected in cider pressed from culled tree-picked apples stored for 4 to 6 weeks at 0 to 2 degrees C but was found at levels of 0.97 to 64.0 microg/liter in cider pressed from unculled fruit stored under the same conditions. Cider from controlled-atmosphere-stored apples that were culled before pressing contained 0 to 15.1 microg of patulin per liter, while cider made from unculled fruit contained 59.9 to 120.5 microg of patulin per liter. The washing of ground-harvested apples before pressing reduced patulin levels in cider by 10 to 100%, depending on the initial patulin levels and the type of wash solution used. These results indicate that patulin is a good indicator of the quality of the apples used to manufacture cider. The avoidance of ground-harvested apples and the careful culling of apples before pressing are good methods for reducing patulin levels in cider.

Beverages↗

Prevalence of Escherichia coli O157 and levels of aerobic bacteria and Enterobacteriaceae are reduced when hides are washed and treated with cetylpyridinium chloride at a commercial beef processing plant.

The objective of this experiment was to test the potential of a combined water wash and cetylpyridinium chloride (CPC) treatment as a hide intervention applied to cattle in the holding pens of a processing plant immediately before stunning. Over 2 processing days, 149 control and 139 treated cattle were tested. Control cattle were processed in the normal manner. The treatment group was prewashed with water the day before harvest. Immediately before stunning, these cattle were sprayed twice with 1% CPC, first for 3 min, then for 1 min. Hides and preevisceration carcasses were sampled to determine aerobic plate counts, Enterobacteriaceae counts (EBC), and Escherichia coli O157 prevalence. The treatment reduced the prevalence of E. coli O157 on hides from 56% to 34% and the prevalence on preevisceration carcasses from 23% to 3%. The treatment decreased aerobic plate counts from 4.9 log CFU/100 cm2 to 3.4 log CFU/100 cm2 and EBC from 3.1 log CFU/100 cm2 to 2.0 log CFU/100 cm2 on preevisceration carcasses. The treatment of hides did not result in any detectable CPC contamination of the chilled carcasses. These data indicated that a 1% CPC treatment preceded by a water wash was capable of reducing hide prevalence of E. coli O157 from as high as 80% to less than 50%, resulting in preevisceration carcass prevalence of 5% or less. We conclude that water washing followed by an antimicrobial treatment, such as CPC, has great potential as an effective hide intervention step and should be further evaluated for implementation as a processing step after stunning and before hide removal.

Abattoirs↗

Quick-staining urinary cytology and bladder wash image analysis with an integrated risk classification: a worthwhile improvement in the follow-up of bladder cancer?

BACKGROUND: With an end toward an increase in patient quality of life, morphologic methods were tested for their combinatory value in expanding the effectiveness of follow-up appointments and finding a more specific supervision of patients with bladder cancer. METHODS: Voided urine and bladder washing specimens were gathered in 223 follow-up sessions of 124 patients with a history of bladder cancer. Hemacolor (Merck, Darmstadt, Germany)-stained cytospin preparations of voided urine specimens were ready for diagnosis within 15 minutes, and results were available shortly before cystoscopy. Feulgen-Schiff-stained cytospin preparations of bladder washings entered the image analysis system. A special software was used to classify the DNA histogram by a risk factor for bladder cancer. RESULTS: Follow-up of patients revealed 83 tumor recurrences. Depending on the grade of the underlying tumor, the sensitivity of quick-staining cytology was 86.4%, 46.2%, or 13.6% for grade 3 to grade 1 TCC, respectively. Cytology and image analysis data demonstrated complementary potency. The combination of methods increased sensitivity to 90.9%, 66.7%, and 31.8%, respectively. Although 24 of 140 image analyses denoted high risk for bladder cancer without simultaneously visible tumor, correct evidence of high risk could be found for 92.2%. CONCLUSIONS: The combinatory use of quick-staining urinary cytology and bladder wash image analysis was demonstrated to be most valuable in diagnosing recurrent bladder cancer and selecting patients needing more intensive follow-up. At a minimum of patients discomfort, the tested combination also seems helpful to surpass diagnostic limits in cystoscopy and cytology caused by therapeutic effects on the bladder epithelium. Cancer (Cancer Cytopathol)

Adolescent↗