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The 24-hour posttransfusion survival, oxygen transport function, and residual hemolysis of human outdated-rejuvenated red cell concentrates after washing and storage at 4 degrees C for 24 to 72 hours.

Red cell concentrates with hematocrit values of 80 +/- 5 percent were stored at 4 degrees C either in citrate-phosphate-dextrose for 22 to 28 days or in citrate-phosphate-dextrose-adenine-one for 35 to 39 days. After storage, the red cells had reduced 2,3-diphosphoglycerate and adenosine triphosphate levels, and biochemical treatment with a solution called PIPA was used to restore these levels. The red cells were not preserved further, but instead were washed after rejuvenation with an unbuffered sodium chloride-glucose solution, pH 5.0, for in vivo studies, and with a buffered sodium chloride-glucose-phosphate solution, pH 6.8, for comparative in vitro studies. The red cells were stored in the wash solution at 4 degrees C for 72 hours after washing. Red cell recovery after washing was about 95 percent. Twenty-four-hour posttransfusion survival value was about 80 percent, and the index of therapeutic effectiveness was greater than 75 percent. These biochemically modified washed red cells exhibited higher than normal P50 values, even after 3 days of postwash storage at 4 degrees C. The units that were washed with the sodium chloride-glucose solution with a pH of 5.0 exhibited a greater degree of hemolysis after 3 days of postwash storage at 4 degrees C than did the units that were washed with the sodium chloride-glucose-phosphate solution with a pH of 6.8.(ABSTRACT TRUNCATED AT 250 WORDS)

2,3-Diphosphoglycerate↗

Two-dimensinal gel electrophoresis of rat liver nuclear washes, nuclear matrix, and hnRNA proteins.

The proteins of rat liver cytoplasm, nuclear washes, matrix, membrane, heterogeneous nuclear (hn)RNA proteins and chromatin were examined by two-dimensional gel electrophoresis. The inclusion in the gels of six common protein standards of carefully selected molecular weight and isoelectric point allowed us to clearly follow the distribution of specific proteins during nuclear extraction. In the nuclear washes and chromatin, we observed five classes of proteins: (a) Exclusively cytoplasmic proteins, present in the first saline-EDTA wash but rapidly disappearing from subsequent washes; (b) ubiquitous proteins of 75,000, 68,000, 57,000, and 43,000 mol wt, the latter being actin, found in the cytoplasm, all nuclear washes and the final chromatin pellet; (c) proteins of 94,000, 25,000, and 20,500 mol wt specific to the nuclear washes; (d) proteins present in the nuclear washes and final chromatin, represented by species at 62,000, 55,000, 54,000, and 48,000 mol wt, primarily derived from the nuclear matrix; and (e) two proteins of 68,000 mol wt present only in the final chromatin. The major 65,000-75,000-mol wt proteins seen by one-dimensional gel electrophoresis of nuclear matrix were very heterogeneous and contained a major acidic, an intermediate, and a basic group. A single 68,000-mol wt polypeptide constituted the majority of the membrane-lamina fraction, consistent with immunological studies indicating that a distinct subset of matrix proteins occurs, associated with heterochromatin, at the periphery of the nucleus. Actin was the second major nuclear membrane-lamina protein. Two polypeptides at 36,000 and 34,000 mol wt constituted 60% of the hnRNP. Approximately 80% of the mass of the nonhistone chromosomal proteins (NHP) from unwashed nuclei is contributed by nuclear matrix and hnRNPs, and essentially the same patterns were seen with chromatin NHP. The concept of NHP being a distinct set of DNA-bound proteins is unnecessarily limiting. Many are derived from the nuclear matrix or hnRNp particles and vary in the degree to which they share different intracellular compartments.

Animals↗

Recovery of ventilation distributions by gas wash-out of a mechanical pump.

A method for deriving virtually continuous distributions of ventilation in the lungs from multiple-breath wash-out of inert, insoluble gases has been tested using a mechanical pump in which two parallel compartments, simulating lung regions, could be differentially ventilated to any desired, and known, extent. With more than moderate non-uniformity, bimodal distributions were always recovered from wash-out data, and with high reproducibility. In a substantial proportion of wash-out experiments ventilation was recovered in regions of very low and very high turnover in addition to the expected modes. These spurious modes may be abolished by various computational devices, none entirely satisfactory. Simultaneous wash-out and wash-in of two or three gases of similar diffusivity give essentially identical solutions. When the pump is operated with the two cylinders out of phase, emptying patterns derived from gas wash-out correspond quite well with those expected from the pump setting. These results help to identify and clarify some of the errors which affect physiological wash-out studies.

Gases↗

Semen characteristics in human immunodeficiency virus (HIV)- and hepatitis C (HCV)-seropositive males: predictors of the success of viral removal after sperm washing.

BACKGROUND: Human immunodeficiency virus (HIV)/hepatitis C virus (HCV)-seropositive males can now father children safely, avoiding transmission risks to the mother and the children using sperm washing and nested PCR (nPCR) techniques. Nevertheless, we still lack enough data to determine the reasons why approximately 10% of the performed sperm washes remain positive, thus forcing the repetition of the treatment. Semen quality in infected males is also essential to these procedures. We aimed to determine the predictive value of the semen parameters, sperm washing procedure and the infection status for the post-wash viral positivity, as well as the correlation between the semen and the disease features. METHODS: Semen characteristics were evaluated in 136 samples provided from 125 males. We also included a control group of 125 males matched by age and length of sexual abstinence. At the time of semen retrieval, 70 of them were infected with HIV (45 also with HCV, 64.3%), and 55 of them with HCV alone. nPCR for viral detection was performed in each sample. RESULTS: Thirteen out of 136 (9.5%) of the samples were positive for one or more viral detections (HIV RNA, HIV DNA and HCV RNA, when needed). From a total of 240 nPCR viral analyses, 16 were positive (6.6%). None of the seminal parameters were adequate to predict post-wash results, nor was a positive result dependent on the volumes used in the semen wash. A positive correlation was found between post-wash progressive motility and CD4 blood levels, as well as a negative correlation between progressive motility and time of evolution of the disease in HIV-infected males. CONCLUSIONS: Semen analysis, according to the World Health Organization criteria, of HIV- and HCV-affected patients showed no differences from that of non-infected males. Moreover, low CD4 blood levels, and a long evolution of the disease do not negatively affect sperm motility.

Anti-Retroviral Agents↗

Peritoneal washing cytology in gynecologic cancers: long-term follow-up of 355 patients.

BACKGROUND: Microscopic evaluation of cells washed from the peritoneal cavity during surgery for gynecologic tumors is used to detect subclinical intraperitoneal metastases from these tumors. The prognostic significance of this test, however, has been questioned. PURPOSE: Stressing histologic correlation and pitfalls in interpretation, we previously reported that the sensitivity of intraoperative peritoneal washing cytology was lower than was suggested earlier. This study evaluates the clinical utility of this test in the long-term follow-up of our patients. METHODS: Staging (International Federation of Gynecology and Obstetrics [FIGO], 1971) and follow-up information was available for 355 unselected patients with primary tumors who had peritoneal washings performed during initial surgery at University Hospital-Stony Brook, NY, during the period from 1980 through 1989. There were 135 patients with endometrial carcinomas, 112 with ovarian carcinomas, 92 with cervical carcinomas, and 16 with borderline (i.e., of low malignant potential) ovarian tumors. The median follow-up of the patients was 57 months (range, 0-154 months). Follow-up data were obtained from the Tumor Registry at University Hospital-Stony Brook. Survival differences were determined by Kaplan-Meier analysis and were evaluated by two-tailed logrank test. RESULTS. Peritoneal washing cytology was positive at initial surgery for 120 (33.8%) of 355 patients, including 90 (80.4%) of 112 patients with ovarian carcinomas, five (31.2%) of 16 patients with borderline ovarian tumors, 17 (12.6%) of 135 patients with endometrial carcinomas, and eight (8.7%) of 92 patients with cervical cancers. For 203 patients with stage I tumors, the peritoneal cytology was positive in 29.4% of the patients with ovarian carcinomas, 18.2% with borderline ovarian tumors, 6.1% with endometrial carcinomas, and 5.2% with cervical carcinomas. By use of peritoneal histology as the standard, peritoneal cytology was highly specific (98.1%) but less sensitive (82.9%) in detecting intraperitoneal involvement. For patients with stage I tumors, 80.0% with ovarian carcinomas, 83.3% with endometrial carcinomas, and 100% with cervical carcinomas who showed positive cytology died of their cancer, compared with 25.0% with ovarian carcinomas, 13.0% with endometrial carcinomas, and 21.9% with cervical carcinomas who showed negative peritoneal cytology. Four (2.0%) patients with stage I tumors had positive peritoneal cytology but negative peritoneal histology. Of these patients, three (two with ovarian carcinoma and one with cervical carcinoma) died of their cancer, whereas one patient with a borderline ovarian tumor was free of disease at the last follow-up. Survival analysis indicated that peritoneal washing cytology stratified for stage provides better prognostic information for each primary cancer site studied than does stage alone. All patients with borderline ovarian tumors were alive at last follow-up, regardless of disease stage or peritoneal status. CONCLUSIONS: Regardless of FIGO stage, positive peritoneal washing cytology predicted poor prognosis for women with epithelial tumors of the genital tract, except for patients with borderline ovarian tumors. Patients in whom peritoneal cytology was the only evidence of intraperitoneal spread were few, but the disease in such patients was associated with poor outcome. IMPLICATIONS: Strict adherence to specialized cytologic criteria in peritoneal washing cytology allows for results that are highly predictive of survival. This information may be useful in stratifying women in therapeutic trials for treatment of genital tract carcinomas.

Adult↗

Isolyte S, a physiologic multielectrolyte solution, is preferable to normal saline to wash cell saver salvaged blood: conclusions from a prospective, randomized study in a canine model.

OBJECTIVES: The purpose of this study is to compare normal saline with Isolyte S as the wash solutions during high-volume cell saver autologous blood transfusion. Normal saline, the standard wash solution in cell saver autologous blood transfusion, is associated with acid-base and electrolyte derangements. Isolyte S is a physiologic, balanced multielectrolyte crystalloid solution that approximates the electrolyte content of plasma. DESIGN: Open-label, prospective, randomized study. SETTING: Research laboratory in a Department of Veterans Affairs medical center. SUBJECTS: Fourteen mongrel dogs, weighing 22 to 23 kg each. INTERVENTIONS: Fourteen mongrel dogs were prospectively randomized to receive normal saline (n = 7) or Isolyte S (n = 7). Animals were anesthetized, received heparin for anticoagulation, and underwent 18 cycles of cell saver autotransfusion. In each cycle, 125 mL of blood was arterially withdrawn, and washed with either normal saline (mEq/L) (sodium 154, chloride 154) or Isolyte S (mEq/L) (sodium 141, potassium 5, magnesium 3, chloride 98, phosphate 1, acetate 28, and gluconate 23). The washed blood was retransfused. MEASUREMENTS AND MAIN RESULTS: Acid-base and electrolyte analyses were performed throughout the study on the systemic blood of each group and compared. By the end of the study, the Isolyte S group had a normal pH and an increased bicarbonate concentration (mEq/L: normal values 24 to 32; normal saline 9.0 +/- 1.9 vs. Isolyte S 13.2 +/- 3.0 [p < .01]) and an increased magnesium concentration (mg/dL: normal values 1.6 to 2.4; normal saline 1.6 +/- 0.2 vs. Isolyte S 2.2 +/- 0.2 [p < .0001]). Additionally, the Isolyte S group had a lower chloride concentration (mEq/L: normal values 95 to 110; normal saline 130 +/- 9 vs. Isolyte S 117 +/- 7 [p < .02]) and a lower potassium concentration (mEq/L: normal values 3.5 to 5.0; normal saline 4.4 +/- 0.5 vs. Isolyte S 3.7 +/- 0.3 [p < .01]). There were no significant differences between normal saline or Isolyte S in the values of PCO2, lactic acid, sodium, total and ionized calcium, inorganic phosphorus, total protein, albumin, hemoglobin, and hematocrit. CONCLUSIONS: Fewer systemic acid-base and electrolyte derangements were observed when blood was washed with Isolyte S. Differences between the normal saline and Isolyte S groups are ascribed primarily to the constituents of the wash solution. We conclude that Isolyte S, a physiologic, balanced, multielectrolyte solution, should be considered as the wash solution in high-volume autologous cell saver blood processing and transfusion.

Animals↗

Value of urethral wash cytology in the retained male urethra after radical cystoprostatectomy.

PURPOSE: We determined the outcome in patients who underwent urethrectomy after cystectomy followed by routine urethral wash cytology versus those not followed by urethral wash cytology who presented with bleeding or urethral discharge. We retrospectively evaluated the outcome in post-cystectomy urethrectomy cases at our institution from 1994 to 2000. MATERIALS AND METHODS: A total of 24 patients with a median age of 70.5 years underwent urethrectomy after cystectomy, including 17 due to asymptomatic, positive urethral wash cytology (group 1) and 7 who were not followed by urethral wash and presented with bleeding/urethral discharge (group 2). Median time from cystectomy to urethrectomy was 11.4 months (range 6.7 to 67.1). Median followup after cystectomy and urethrectomy was 37 and 27.7 months, respectively. RESULTS: Urethrectomy pathological evaluation showed pTis disease in cases 12 (50%), pT0 in 9 (37.5%) and pT1 in 3 (12.5%). Cystectomy pathology was organ confined (pT0, pTis and pT1-pT2b disease) in 12 cases (50%), nonorgan confined (pT3a-pT4) in 6 (25%) and pT any N1 in 5 (21%). Cystectomy pathology was unknown in 1 case. At the most recent followup there was no evidence of disease in 14 patients (58%), 5 (21%) were alive with disease, 3 (12.5%) were dead of disease, 1 (4%) was dead of other causes and disease status was unknown in 1 (4%). There was no statistical difference in survival in groups 1 and 2 when controlling for original bladder tumor stage. Cox regression analysis revealed that cystectomy pathology was the only statistically significant parameter of disease-free survival (p = 0.011), while urethrectomy pathology and followup method (urethral washing versus no washing) were not significant. There were no perioperative or postoperative complications and no patients died. CONCLUSIONS: There was no significant survival difference in patients followed and not followed with urethral washing. Longer followup and increased patient numbers are needed to determine the significance of these findings.

Aged↗

Pulmonary endothelial permeability is increased by fluid from packed red blood cell units but not by fluid from clinically-available washed units.

BACKGROUND: Massive transfusions are a risk factor for acute respiratory distress syndrome (ARDS) in severely injured patients. Neutrophil priming has been proposed to be an integral part of the early inflammatory response to trauma. To complement that work, we studied another major cell type involved in inflammation: the endothelial cell. Our hypothesis was that soluble factors from units of leukoreduced packed red blood cells (PRBC) directly increase pulmonary endothelial permeability. We also determined whether fluid from clinically-available washed PRBC units affects endothelial permeability. METHODS: As a measure of permeability, transendothelial electrical resistance (TER) was determined across monolayers of a human pulmonary microvascular endothelial cell line after addition of full-strength, diluted, and washed PRBC fluid. Monolayers were stained with phalloidin to assess intercellular space. Storage solution Adsol-1 was tested alone to determine additive component effects on TER. RESULTS: PRBC fluid decreased TER and increased intercellular space, both of which indicate an increase in endothelial monolayer permeability. PRBC fluid diluted to 2% and washed PRBC fluid did not decrease TER and thereby did not change endothelial permeability. Likewise, Adsol-1 did not duplicate the dramatic decrease in TER seen with the PRBC fluid. CONCLUSIONS: Fluid from stored PRBC units contains a soluble, transferable factor that directly increases endothelial permeability. Fluid from washed PRBC units, currently available for patients with immunoglobulin A allergies, does not. This study complements previous work of others that demonstrated that neutrophil priming by PRBC fluid is abrogated by washing. Now that two cell types have been shown to respond more favorably to washed PRBC in vitro, clinical studies should be initiated to investigate whether use of washed PRBC reduces ARDS following transfusions in trauma patients.

Capillary Permeability↗

Suberization: inhibition by washing and stimulation by abscisic Acid in potato disks and tissue culture.

Wounding of potato (Solanum tuberosum L.) tubers results in suberization, apparently triggered by the release of some chemical factor(s) at the cut surface. Suberization, as measured by diffusion resistance of the tissue surface to water vapor, was inhibited by mm concentrations of indoleacetic acid, unaffected by mm concentrations of traumatic acid, severely inhibited at mum concentrations of cytokinin, but stimulated by abscisic acid (ABA) at 10(-4)m. Thorough washing of potato disks up to 3 to 4 days after cutting resulted in severe inhibition of suberization as measured both by diffusion resistance and by the amount of the octadecene diol generated by hydrogenolysis (LiAlH(4)) of the tissue. Disks washed after 4 days did not show any inhibition of suberization. High performance liquid chromatographic analysis of the wash from fresh potato disks showed that about 14 ng of ABA was released into the wash per g of tissue. The amount of ABA released increased with time up to 4 to 6 hours of washing. The maximal amount of ABA was washed out after aging for 24 hours and after 2 days of aging ABA could no longer be found in the surface wash of the disks. Addition of ABA to the media of potato tissue cultures resulted in suberin formation whereas control cultures contained little suberin. The effect of ABA on suberization in the tissue cultures was shown to be linearly concentration-dependent up to 10(-4)m and a linear increase in suberin formation was seen up to about 8 days of culture growth on the media containing 10(-4)m ABA. From these results it is proposed that during the early phase of wound-healing ABA plays a role in triggering a chain of biochemical processes which eventually (in about 3 to 4 days) result in the formation of a suberization-inducing factor, responsible for the induction of the enzymes involved in suberin biosynthesis.

Journal Article↗

Helicobacter pylori may be transmitted through gastrofiberscope even after manual Hyamine washing.

Endoscopy is an effective diagnostic technique for gastric cancer, which is believed to be associated with Helicobacter pylori. Manual Hyamine washing is a widely used fiberscope cleaning method. Urease B gene of Helicobacter pylori was detected in 50% of the wash-out samples from the biopsy-suction channel of a fiberscope after manual Hyamine washing by nested polymerase chain reaction, and bacterial culture revealed viable Helicobacter pylori in 19%. However, Helicobacter pylori was not detected by either of the above methods in the biopsy-suction channel of the fiberscope after mechanical washing. These findings indicate that manual Hyamine washing of fiberscopes is insufficient to prevent iatrogenic Helicobacter pylori transmission, and that mechanical washing after manual Hyamine washing is essential.

Base Sequence↗

Plasma-depleted platelet concentrates prepared with a new washing solution.

In certain clinical situations, complete removal of the plasma proteins from the platelet concentrates (PCs) is necessary by washing prior to transfusion. A simple electrolyte solution with a pH of 6.5 was developed for washing PCs. The platelet-rich plasma collected with acid-citrate-dextrose solution by apheresis in a 0.6-liter polyolefin bag was centrifuged. After removal of the supernatant plasma from pelleted platelet buttons, 200 ml of a washing solution consisting of 90 mM NaCl, 5 mM KCl, 3 mM MgCl2, 17 mM NaH2PO4, 8 mM Na2HPO4, 23 mM Na acetate, 17 mM Na3 citrate, 23.5 mM glucose, 2 mM adenine, 0.1% dextran, and 28.8 mM maltose (pH 6.5) was added to the pelleted platelet button. Steam sterilization of the solution was carried out under nitrogen to avoid caramelization of glucose. After resuspension of the pelleted platelet button with a washing solution and a second centrifugation, Seto additive solution (Seto sol, pH 7.4) was introduced into the bag to resuspend the platelet buttons for storage for 3 days at 22 degrees C. All of these procedures were completed within 3 h using a sterile docking device. In washed PCs, 99.1% of the plasma was removed and platelet recovery was 96%. The washed PCs were compared for 3 days with plasma-poor PCs consisting of 11% plasma and 89% Seto solution. There were no significant differences in percent hypotonic shock response, aggregation, energy metabolism, and morphology of platelets between the two groups during 3 days, except for significant swelling of 3-day-old platelets in washed PCs.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗

Effects of different staining and washing procedures on the results of human sperm morphology evaluation by manual and computerised methods.

The purpose of this study was to investigate the effect of different staining and washing procedures on the results of human sperm morphology evaluation by manual and computerised methods. Furthermore, it was intended to find the staining and washing combination which would provide optimal readability for computer-assisted sperm morphology evaluations. In phase one, four staining methods were evaluated for smears prepared from the resulting samples following a two times washing procedure. In phase two, 20 semen samples were used to compare the Diff-Quik and Papanicolaou staining methods, following one and two washes. All manual readings, of Papanicolaou and Diff-Quik stained smears, were comparable with each other, with means between 7.3% and 7.9% normal spermatozoa. All the manual readings were also comparable to the computer readings of the Diff-Quik slides following one and two washes with means of 9.0% and 5.9%, respectively. However, due to the higher computer readings found for the Papanicolaou stained smears, with means of 13.9% and 13.5% following one and two washes, respectively, a statistically significantly difference between overall computer and manual readings was found (Wilks' Lamda, P = 0.0002). Taking all data into consideration, it could be concluded that the one wash Diff-Quik stained smears was the optimal preparation method for computerised sperm morphology evaluation, comparing favourably with manual evaluations.

Evaluation Studies as Topic↗

The use of bladder wash-outs to reduce urinary catheter encrustation.

OBJECTIVE: To examine the effectiveness of bladder washouts in reducing catheter encrustation. MATERIALS AND METHODS: A model of the catheterized bladder was developed and used to produce catheter encrustation in vitro. The effects of bladder wash-outs with Suby G, mandelic acid 1% or saline 0.9% were compared with control catheters that did not receive wash-outs, by measuring the area of patent catheter lumen and the changes in urinary calcium and magnesium concentrations after wash-outs. RESULTS: Encrustation was reduced significantly, compared with the controls, in catheters washed out with the acidic reagents, Suby G and mandelic acid. Saline wash-outs did not reduce encrustation. CONCLUSION: The model of the catheterized bladder provided a reliable technique with which to examine the effects of bladder wash-out on catheter encrustation under controlled conditions. The results indicate that bladder wash-outs with Suby G or mandelic acid may be beneficial for patients who suffer from recurrent catheter encrustation and blockage.

Calcium↗

Gender difference of in vivo skin surface pH in the axilla and the effect of a standardized washing procedure with tap water.

The purpose of our study was to evaluate the axillary skin surface pH and explore potential gender-related differences together with the influence of a washing procedure in healthy subjects. After a run-in period, the skin surface pH was measured in vivo in 10 men and 10 women under standardized conditions in three distinct locations of each axilla (at baseline and up to 6 h after washing). Potential interfering influences were thoroughly excluded. Our study revealed a statistically significant difference in axillary skin surface pH between men and women with more acidic values in females (p < or = 0.001). The baseline axillary pH before washing was 6.58 +/- 0.63 (right) and 6.67 +/- 0.65 (left) in men compared to 5.8 +/- 0.53 (right) and 5.94 +/- 0.62 (left) in women. The difference between the right and left arm-pit was not statistically significant (p > 0.05) in any group. After standardized washing procedures with pure tap water, the mean axillary pH decreased significantly in women at all measurement times [lowest value 60 min after washing: 5.51 +/- 0.7 (right) and 5.64 +/- 0.7 (left)], while it slightly increased in men [highest value 240 min after washing: 6.7 +/- 0.59 (right) and 6.78 +/- 0.69 (left)]. In summary, there is a gender difference in axillary skin surface pH. Washing with water further increased the difference between male and female pH values.

Adult↗

Sensitive identification of mycobacterial species using PCR-RFLP on bronchial washings.

In 98 patients (24 with active pulmonary tuberculosis [TB] lesions, 28 with cured TB lesions, and 46 with nontuberculous opacities [control group] in chest CT scans), we examined whether washing the bronchus after brushing the lesion, then applying polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to the bronchial washings might be useful for diagnosing TB and nontuberculous mycobacteriosis (NTMosis). After biopsy and brushing with a bronchoscope, the bronchus connecting to the lesion was washed with 20 ml saline. The saline used for washing the brushes (5 ml; brushing sample), and 3 to 10 ml saline aspirated through the forceps channel (washing sample) were examined by PCR-RFLP, which proved able to identify Mycobacterium tuberculosis and seven species of nontuberculous mycobacteria (NTM). The values obtained for the sensitivity of the PCR-RFLP with respect to the brushing sample, the washing sample, and both samples mixed together were 70, 76, and 91%, respectively, when only patients who were culture-positive or radiologically improved after antituberculous therapy were considered as showing true infection. A mixture of brushing and washing samples provides useful material for PCR and culture, and the PCR-RFLP used here is a good method for the simultaneous identification of several species of mycobacterium (including M. tuberculosis).

Adult↗

Bacteriology of expectorated sputum with quantitative culture and wash technique compared to transtracheal aspirates.

Techniques to improve the reliability of expectorated sputum culture were evaluated in 46 patients using a wash technique, quantitative culture, and a combination of washing and quantification. The wash procedure consisted of a jet of tap water over the specimen contained in a tea strainer. The true lower respiratory tract flora was determined by percutaneous transtracheal aspirates, which served as the basis for evaluating the methods of processing expectorated specimens. Either washing alone or quantification alone decreased the number of specimens with organisms that were not present in companion transtracheal aspirates; however, results that were sufficiently improved to be clearly advantageous for clinical interpretation were achieved only when washing and quantification were combined. Washed sputum specimens yielded oropharyngeal "contaminants" in concentrations exceeding 10(6) bacteria per ml in 12 of 46 specimens (26 per cent). Using this technique, only one specimen contained a misleading potential pathogen, and only one specimen failed to yield a potential pathogen that was recovered with the transtracheal aspirate. The wash procedure decreased the mean concentrations of contaminants approximately 100-fold for all specimens and 1,000-fold for purulent specimens.

Anaerobiosis↗

Residual protein contamination of ProSeal laryngeal mask airways after two washing protocols.

The risk of prion protein cross-infection has focussed attention on the potential hazards of protein contamination of re-usable medical devices. This study determined the frequency of protein contamination of ProSeal laryngeal mask airways (PLMA) after two cleaning procedures and tested the hypothesis that the combination of hand- and machine-washing removes protein contamination more effectively than hand-washing alone. After clinical use fifty-four PLMAs were randomly allocated to be washed by hand or by hand then machine. All PLMAs were then autoclaved at 134 degrees C for 4 minutes. After processing, each PLMA was immersed in a 1.2% solution of erythrosin B and examined for uptake of stain. The site (outer surface, bowl and edges of the cuff airway and drain tube, finger strap) and severity (nil/mild/moderate/severe) of staining was scored by a blinded observer. There were no differences in the site or severity of staining between the two cleaning procedures. Staining was detected on 89% of PLMAs that were hand-washed and 78% of PLMAs that were hand-, then machine-washed (P=0.27). When staining occurred, it was mild in 98%, moderate in 2% and was never severe. Staining was more frequent on the edge than at any other location (all comparisons: P < or = 0.01). The strap never had any staining. We conclude that residual contamination of PLMAs with protein deposits is common even when machine-washing is used to augment hand-washing before autoclaving. The infection risk associated with these deposits remains to be determined.

Cross Infection↗

Does a preceding hand wash and drying time after surgical hand disinfection influence the efficacy of a propanol-based hand rub?

BACKGROUND: Recently, a propanol-based hand rub has been described to exceed the efficacy requirements of the European standard EN 12791 in only 1.5 min significantly. But the effect of a 1 min preceding hand wash and the effect of one additional minute for evaporation of the alcohol after its application on the efficacy after a 1.5 min application time has never been studied. METHODS: We have investigated a propanol-based hand rub (Sterillium, based on 45% propan-2-ol, 30% propan-1-ol and 0.2% mecetronium etilsulfate) in three variations: with (A) and without (B) a 1 min hand wash before the disinfection of 1.5 min with immediate sampling after the disinfection; and (C) without a hand wash before the disinfection but with sampling 1 min after the disinfection. The efficacy of the three variations was compared to the reference treatment of EN 12791. All experiments were performed in a Latin-square design with 20 volunteers. Pre- and post-values (immediate and after 3 hr) were obtained according to EN 12791. RESULTS: The 3 min reference disinfection reduced resident hand bacteria on average by 1.8 log10 steps (immediate effect) and 1.4 log10-steps (sustained effect) respectively. Method A was equally effective as the reference (immediate efficacy: 1.5 log10-steps; sustained efficacy: 1.6 log10-steps). Method B seemed to be more effective (immediate efficacy: 2.3 log10-steps; sustained efficacy: 1.7 log10-steps). Method C revealed the best efficacy (immediate efficacy: 2.3 log10-steps; sustained efficacy: 2.0 log10-steps). A comparison of all three treatment variations and the reference treatment revealed a significant difference for the immediate efficacy (p = 0.026; Friedman test), but not for the sustained efficacy (p = 0.430). A post-hoc-test for the immediate efficacy indicated a significant difference between methods A and C (p < 0.05; Wilcoxon-Wilcox test). Hence, none of the treatment variations was significantly less effective than the reference treatment. CONCLUSION: An application of the propanol-based hand rub for 1.5 min after 1 min hand wash fulfills the efficacy requirements of EN 12791. The efficacy can be improved to some extent by omitting the preceding hand wash and by awaiting the evaporation of the alcohol which is clinical practice anyway. The preceding hand wash has the most negative effect on the immediate effect. Based on our data hands should not be routinely washed before the disinfection period unless there is a good reason for it such as visible soiling.

Anti-Infective Agents, Local↗