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Effects of washing with a neutralizing agent on alkaline skin injuries in an experimental model.

In this report a skin alkaline injury model was constructed using experimental rats and 2 N NaOH. We observed the effects of washing with a neutralizing agent on the subcutaneous tissue pH at 1, 10 and 30 min after injury and compared it with water-washing groups. In comparison with the water washing group, the peak pH values following the use of the neutralizing agent were significantly higher in the 1-min washing group, and significantly lower in the 10-min and 30-min washing groups. Changes in the pH values of the subcutaneous tissue after reaching a peak showed a similar course in the 1-min washing group, whereas in the 10 and 30 min washing groups there was a significantly different rate of decrease of pH. These results indicate that although treatment with a neutralizing agent directly after injury causes harmful effects due to the neutralizing reaction with a highly concentrated alkali. Subsequently with prolonged washing there is an effective reduction of the high pH.

Animals↗

Evaluation of different techniques for washing cats: quantitation of allergen removed from the cat and the effect on airborne Fel d 1.

BACKGROUND AND OBJECTIVE: The purpose of this study was to examine the quantity and distribution of the major cat allergen, Fel d 1, on cats and to evaluate the efficacy of washing, both in removing allergen from the cat and reducing airborne allergen levels. METHODS: Airborne samples were collected on four glass fiber filters in a 30 m3 room, before and 3 hours after serial washing of eight cats (45-minute sampling at 18 L/min for each filter). Aliquots of hair and bath water were also collected and assayed for Fel d 1 content. RESULTS: Extracting cat hair with tap water or pet shampoo for 3 minutes removed mean levels of 191 and 245 microg of Fel d 1 per gram of hair, respectively; the quantity of allergen on samples of cat hair ranged from 1 microg/gm to more than 1770 microg/gm. The highest concentration of allergen was found on hair from the neck. Estimates of the total Fel d 1 on the cat, based on shaving the whole cat, ranged from 3 to 142 mg (mean = 67 mg). Washing cats reduced airborne allergen 3 hours later. Washing three cats at weekly intervals for 5 weeks in a veterinarian's office produced a mean decrease of 44% in airborne Fel d 1 (n = 15, p < 0.02). Washing three cats by immersion for 3 minutes at weekly intervals for a 1-month period produced a mean decrease in airborne allergen of 79% (n = 12, p < 0.001). However, after repeated washing, the airborne levels before the next wash were not consistently decreased. The quantity of Fel d 1 removed by immersion varied from 1 to 35 mg. CONCLUSION: Cats carry large quantities of Fel d 1, only a small proportion of which (approximately 0.002%/hr) becomes airborne. Washing cats by immersion will remove significant allergen from the cat and can reduce the quantity of Fel d 1 becoming airborne. However, the decrease is not maintained at 1 week.

Air Pollutants↗

Risk assessment of hand washing efficacy using literature and experimental data.

This study simulated factors that influence the levels of bacteria on foodservice workers' hands. Relevant data were collected from the scientific literature and from laboratory experiments. Literature information collected included: initial bacterial counts on hands and water faucet spigots, bacterial population changes during hand washing as effected by soap type, sanitizing agent, drying method, and the presence of rings. Experimental data were also collected using Enterobacter aerogenes as a surrogate for transient bacteria. Both literature and experimental data were translated into appropriate discrete or probability distribution functions. The appropriate statistical distribution for each phase of the hand washing process was determined. These distributions were: initial count on hands, beta (2.82, 2.32, 7.5); washing reduction using regular soap, beta (3.01, 1.91, -3.00, 0.60); washing reduction using antimicrobial soap, beta (4.19, 2.99, -4.50, 1.50); washing reduction using chlorhexidine gluconate (CHG), triangular (-4.75, -1.00, 0); reductions from hot air drying, beta (3.52, 1.92, -0.20, 1.00); reduction from paper towel drying, triangular (-2.25, -0.75, 0); reduction due to alcohol sanitizer, gamma (-1.23, 4.42) -5.8; reduction due to alcohol-free sanitizer, gamma (2.22, 5.38) -5.00; and the effect of rings, beta (8.55, 23.35, 0.10, 0.45). Experimental data were fit to normal distributions (expressed as log percentage transfer rate): hand-to-spigot transfer, normal (-0.80, 1.09); spigot to hand, normal (0.36, 0.90). Soap with an antimicrobial agent (in particular, CHG) was observed to be more effective than regular soap. Hot air drying had the capacity to increase the amount of bacterial contamination on hands, while paper towel drying caused a slight decrease in contamination. There was little difference in the efficacy of alcohol and alcohol-free sanitizers. Ring wearing caused a slight decrease in the efficacy of hand washing. The experimental data validated the simulated combined effect of certain hand washing procedures based on distributions derived from reported studies. The conventional hand washing system caused a small increase in contamination on hands vs. the touch-free system. Sensitivity analysis revealed that the primary factors influencing final bacteria counts on the hand were sanitizer, soap, and drying method. This research represents an initial framework from which sound policy can be promulgated to control bacterial transmission via hand contacts.

Bacteria↗

Can patients wash during radiotherapy to the breast or chest wall? A randomized controlled trial.

A total of 99 patients receiving adjuvant radiotherapy to the breast or chest wall were randomized to one of three washing policies: (1) not washing, (2) washing with water alone, and (3) washing with soap and water. Fifty-three of the patients were treated without the use of bolus, and 46 patients were treated using bolus for 10 to 15 of the 20 treatment fractions. Approximately half of the patients randomized to washing were tattooed. Subjective and objective measurements of acute skin reactions were found to be less in the two groups randomized to washing. There was little difference between the two washing groups. These findings were the same whether or not bolus was used. It is concluded that washing of the skin should be encouraged in patients undergoing radiotherapy associated with low skin doses.

Baths↗

Efficacy of a hot washing process for pretreated yellow poplar to enhance bioethanol production.

Cost reductions for pretreatment and bioconversion processes are key objectives necessary to the successful deployment of a bioethanol industry. These unit operations have long been recognized for their impact on the production cost of ethanol. One strategy to achieve this objective is to improve the pretreatment process to produce a pretreated substrate resulting in reduced bioconversion time, lower cellulase enzyme usage, and/or higher ethanol yields. Previous research produced a highly digestible pretreated yellow poplar substrate using a multistage, continuously flowing, very dilute sulfuric acid (0.07% (w/v)) pretreatment. This process reduced the time required for the bioconversion of pretreated yellow poplar sawdust to ethanol. This resulted in a substantially improved yield of ethanol from cellulose. However, the liquid volume requirements, steam demand, and complexity of the flow-through reactor configuration were determined to be serious barriers to commercialization of that process. A reconfigured process to achieve similar performance has been developed using a single-stage batch pretreatment followed by a separation of solids and liquids and washing of the solids at a temperatures between 130 and 150 degrees C. Separation and washing at the elevated temperature is believed to prevent a large fraction of the solubilized lignin and xylan from reprecipitating and/or reassociating with the pretreated solids. This washing of the solids at elevated temperature resulted in both higher recovered yields of soluble xylose sugars and a more digestible pretreated substrate for enzymatic hydrolysis. Key operating variables and process performance indicators included acid concentration, temperature, wash volume, wash temperature, soluble xylose recovery, and performance of the washed, pretreated solids in bioconversion via simultaneous saccharification and fermentation (SSF). Initial results indicated over a 50% increase in ethanol yield at 72 h for the hot washed material as compared to the control (no washing, no separation) and a 43% reduction of in the bioconversion time required for a high ethanol yield from cellulose

Biomass↗

Limited impact of sustained simple feedback based on soap and paper towel consumption on the frequency of hand washing in an adult intensive care unit.

OBJECTIVE: To determine whether hand washing would increase with sustained feedback based on measurements of soap and paper towel consumption. DESIGN: Prospective trial with a nonequivalent control group. SETTING: Open multibed rooms in the Omaha Veterans Affairs Medical Center's Surgical Intensive Care Unit (SICU) and Medical Intensive Care Unit (MICU). SUBJECTS: Unit staff. INTERVENTION: Every weekday from May 26 through December 8, 1998, we recorded daytime soap and paper towel consumption, nurse staffing, and occupied beds in the SICU (intervention unit) and the MICU (control unit) and used these data to calculate estimated hand washing episodes (EHWEs), EHWEs per occupied bed per hour, and patient-to-nurse ratios. In addition, from May 26 through June 26 (baseline period) and from November 2 through December 8 (follow-up period), live observers stationed daily for random 4-hour intervals in the MICU and the SICU counted actual hand washing episodes (CHWEs). The intervention consisted of posting in the SICU, but not in the MICU, a graph showing the weekly EHWEs per occupied bed per hour for the preceding 5 weeks. RESULTS: Directly counted hand washing fell in the SICU from a baseline of 2.68+/-0.72 (mean +/- standard deviation) episodes per occupied bed per hour to 1.92+/-1.35 in the follow-up period. In the MICU, episodes fell from 2.58+/-0.95 (baseline) to 1.74+/-0.69. In the MICU, the withdrawal of live observers was associated with a decrease in estimated episodes from 1.36+/-0.49 at baseline to 1.01+/-0.36, with a return to 1.16+/-0.50 when the observers returned. In the SICU, a similar decrease did not persist throughout a period of feedback. Estimated hand washing correlated negatively with the patient-to-nurse ratio (r = -0.35 for the MICU, r = -0.46 for the SICU). CONCLUSIONS: Sustained feedback on hand washing failed to produce a sustained improvement. Live observers were associated with increased hand washing, even when they did not offer feedback. Hand washing decreased when the patient-to-nurse ratio increased.

Adult↗

Effects of applying Safe2O poultry wash to broiler wings on shelf life, Listeria monocytogenes, Pseudomonads, Staphylococcus species, and psychrotrophic bacteria levels after three, seven, and ten days of storage.

Bacterial contamination of raw processed poultry continues to be of concern to consumers as well as regulatory and health officials. For many years wings were considered a low-value product; therefore, shelf life of wings was not a major concern. Due to changes in consumer attitudes and increases in the fast-food market, wings are now a valuable commodity. Because wings have a shorter shelf life than most other raw poultry products, acceptable intervention to decrease the population of associated spoilage organisms and human enteropathogens are needed. Safe2O Poultry Wash was evaluated as a postchill treatment to reduce microbial contamination and increase shelf life. Ninety-six carcasses were obtained from a local processor prior to final wash. On arrival at the research facility all carcasses were inoculated with 1 mL of a culture with 10(3) cfu/mL Listeria monocytogenes. After a 30-min attachment time, carcasses were subjected to a 4-s in-out final wash, hung for 3 min, and chilled in ice-water for 45 min. After the chilling, wings were removed by hand with a knife, pooled together, and subjected to a hand spray (4 mL/wing) with deionized water or Safe2O Poultry Wash. Two wings were then placed in each of 96 ziplock type storage bags, and wings were held at 5 +/- 1 degrees C for 3, 7, 10, and 14 d. On the day of sample, weep was decanted, and 100 mL of Butterfield's phosphate buffer was added to each bag. Three sets of wings were shaken by hand for 1 min, and total aerobes, Pseudomonads, Staphylococcus sp., psychrotrophic bacteria, and L. monocytogenes in the rinsates were enumerated. By using 7 log10 recovery of total aerobes from rinsates as a spoilage baseline, all wings were spoiled by d 10, but the wings treated with water were approaching spoilage counts on d 7, (log10 6.8), whereas only log10 5.5 bacteria were recovered from the wings sprayed with Safe2O Poultry Wash. Fewer Pseudomonads, Staphylcoccus sp., L. monocytogenes, and psychrotrophic bacteria were recovered from wings treated with Safe2O Poultry Wash and stored for 10 d. Log10 counts for the organisms were Pseudomonas sp., 8.2 and 6.9; Staphylcoccus sp., 5.5 and 4.9; L. monocytogenes, 5.2 and 4.6; and psychrotrophs, 8.2 and 6.9 for the water and Safe2O Poultry Wash treatments, respectively. Use of the Safe2O Poultry Wash as a postchill treatment on wings could increase the shelf life of wings by up to 3 d.

Animals↗

Increase of IgA in pharyngeal washings from patients with IgA nephropathy.

The levels of IgA and total proteins in pharyngeal washings were examined in patients with IgA nephropathy and other glomerular diseases in order to determine whether secretory IgA, IgA1 and IgA2 were increased in these patients. Thirty patients with IgA nephropathy, 21 patients with other glomerular diseases and 15 healthy adults were examined. The levels of IgA were quantitated by radial immunodiffusion and laser nephelometry. The levels of total proteins in pharyngeal washings were quantitated by Tonein-TP. It was demonstrated that the levels of IgA in pharyngeal washings were significantly increased in patients with IgA nephropathy compared to those with other glomerular diseases and healthy adults. The levels of total proteins in pharyngeal washings were increased in patients with IgA nephropathy, and the ratio of IgA and total proteins in the washings from patients with IgA nephropathy was not significantly different from those healthy adults. There was a significant correlation between the levels of IgA in pharyngeal washings and those of IgA in sera from patients with IgA nephropathy. The IgA observed in pharyngeal washings contained secretory components, IgA1 and IgA2, in patients with IgA nephropathy. It is suggested that the increase of IgA in the pharyngeal washings in parallel with the increase of total proteins might be associated with local inflammation.

Humans↗

Breast stereotactic core biopsy washings: abundant cell samples from clinically occult lesions for flow cytometric DNA analysis.

RATIONALE AND OBJECTIVES: The object of this study was to determine by flow cytometry the cellular and DNA content of the washings of core biopsy samples and needles obtained during stereotactic core biopsies of clinically occult suspicious lesions. METHODS: Fourteen-gauge core multipass biopsy inner-needle saline washings obtained after dislodgment of the tissue core samples (n = 30) and 14-ga mammotomy core biopsy specimen washings (n = 30) were analyzed by flow cytometry. RESULTS: Multipass core biopsy needle washings of 7 malignant and 23 benign lesions yielded a median of 10,584 cells per lesion (range 562-29,019 cells). Mammotomy core biopsy specimen washings of 10 malignant and 20 benign lesions yielded a median of 12,164 cells per lesion (range 1,295-115,150). Forty-four (73%) washings in each technique had more than 5000 cells. Flow cytometric DNA ploidy analysis was possible in 60 (100%) and the S phase percentage determination was possible in 59 (98%) of the washings. Aneuploidy was detected in 53% of malignant lesions and 2% of benign lesions. CONCLUSIONS: Core biopsy specimen or needle washings provide abundant fresh cellular material from clinically occult lesions for DNA analysis by flow cytometry.

Adult↗

Tolerance and safety of different concentrations of chlorhexidine for peripartum vaginal and infant washes: HIVNET 025.

BACKGROUND: There is a continuing need to evaluate sustainable interventions for prevention of mother-to-child transmission (MTCT) of HIV type 1. We evaluated different concentrations (0.25%, 1%, and 2%) of chlorhexidine (CHX) for perinatal maternal and infant washes to identify the maximum tolerable concentration of CHX for such an intervention. METHODS: Women were enrolled during their third trimester at the maternity unit of the Chris Hani Baragwanath Hospital in Soweto, South Africa, and perinatal maternal and infant washes were completed. Subjective maternal symptoms as well as infant examinations were used to assess tolerability of the washes. RESULTS: The 0.25% concentration of CHX was well tolerated by the mothers (n = 29). Ten of 79 women (13%) with 1% CHX washes complained of mild vaginal area burning or itching, and washes were stopped in 5 (6%). Twenty-three of 75 women (31%) in the 2% CHX wash group had subjective complaints, and the washes were stopped in 12 (16%). There were no clinical indications of toxicity of the CHX washes among infants. CONCLUSION: A 1% solution of CHX appears to be a safe and tolerable concentration of CHX for consideration in an MTCT prevention trial.

Acquired Immunodeficiency Syndrome↗

Mite control with low temperature washing. I. Elimination of living mites on carpet pieces.

BACKGROUND: Washing of delicate textiles at low temperature combined with mite control is one of the important aims for domestic mite prevention. OBJECTIVES: Estimation or assessment of mite numbers without and after the application of detergents or detergents with mite control additive. Trials at low temperature, detergents and washing as usual at household conditions (programme of a household washing machine). METHODS: The domestic mite Dermatophagoides farinae can be cultured on carpet samples under favourable conditions, and the breeding success monitored in terms of population and distribution using the free mite mobility test. The mite containing carpets were cut into pieces in order to investigate the effects of washing at low temperature with different commercial detergents with and without an additive for mite control. The heat escape method was used to assess the number of mites remaining on the carpet pieces. RESULTS: The estimation of the mite populations on different carpet pieces before and after washing showed that between 40 and 60% of the mites were able to resist washing, rinsing, spinning and drying. The allergen containing dust, however, was flushed out. In contrast, complete mite control was achieved by the application of the same detergents together with an additive. CONCLUSION: Mite control by low temperature washing is possible with a mite control additive provided that a concentration of benzyl benzoate of about 0.03% is achieved in the washing suds.

Animals↗

P53 and ploidy assessed by flow cytometry in bladder washings.

OBJECTIVE: To determine p53 expression in cells in bladder washings and to relate this to DNA content and clinical outcome. PATIENTS AND METHODS: Washings from 102 patients (41 with newly diagnosed superficial tumours [pTa and pT1], 49 with recurrent superficial tumours and 12 with carcinoma invading bladder muscle) were studied. In 39 cases, the primary bladder tumour was also analysed. The rates of tumour recurrence and progression were determined for the new superficial tumours and related to both p53 expression and DNA content. RESULTS: Cells positive for p53 were detected in 22 of 90 (24%) washings from patients with superficial bladder cancer. P53 expression correlated with tumour stage (P < 0.05), grade (P < 0.05) and abnormal DNA content (P < 0.05). The analysis of pure urothelial (cyto-keratin-positive) cells improved the detection of DNA abnormalities (P < 0.001). In 74% of cases where both washings and tumour were analysed, the results for DNA content agreed. Of 41 new superficial tumours, 27 (66%) recurred (11 were p53-positive, 16 were p53-negative, P = 0.221; 17 had abnormal DNA content, 10 were diploid, P = 0.069). Four patients progressed (one was p53-positive, P = 0.315 and all had abnormal DNA content, P = 0.072). CONCLUSION: P53-positive cells can be detected in washings using flow cytometry and were more commonly detected in association with aneuploid tumours. At short-term follow-up, flow cytometric analysis of DNA content in washings had greater predictive value than had p53 expression. Few washings contained aneuploid cells when the primary tumour contained diploid cells, although the collection of washings is a convenient way of sampling tumour cells.

Aged↗

Wash-out of the non-heart-beating donor liver: a matter of flush solution and temperature?

BACKGROUND AND AIMS: Ischemically damaged donor livers are prone to graft non-function. This can in part be explained by a suboptimal wash-out during procurement. An enriched machine perfusion (MP) preservation solution for livers, named Polysol, was developed. The aim of this study was to investigate the type of flush solution, temperature and anticoagulant content on the wash-out of the non-heart-beating donor (NHBD) rat liver. METHODS: Rat livers were flushed after 30 min warm ischemia. After excision, livers were reperfused at 37 degrees C, with analysis of damage and function, concerning (1) solutions (University of Wisconsin (UW), histidine-tryptophan-ketoglutarate (HTK) and Polysol); (2) temperature (4 degrees C, 18 degrees C and 37 degrees C); (3) addition of heparin and (4) wash-out followed by 24 h MP. RESULTS: (1) Reperfusion results were inferior in the UW group; (2) less damage and improved function were seen after wash-out using Polysol at 37 degrees C; (3) No effects were seen of the addition of heparin to Polysol; (4) MP after wash-out using HTK resulted in more liver damage and decreased liver function as compared with wash-out using Polysol. CONCLUSIONS: Polysol is applicable as a flush solution for the NHBD liver, resulting in equal to better wash-out as compared with UW and HTK. The best temperature for this NHBD wash-out is 37 degrees C.

Adenosine↗

A single-blinded, randomized, controlled clinical trial evaluating the effect of face washing on acne vulgaris.

Despite the common recommendation to wash the face twice daily with a mild cleanser, there is little published evidence to support the practice. Indeed, while the general public believes that cleaner skin will result in fewer blemishes, dermatologists often warn that overwashing and scrubbing can exacerbate the condition. To clarify the effect of frequency of face washing on acne vulgaris, we designed a single-blinded, randomized, controlled clinical trial to be conducted on males with mild to moderate acne vulgaris. Subjects washed their faces twice daily for 2 weeks with a standard mild cleanser before being randomized to one of three study arms, in which face washing was to be done once, twice, or four times a day for 6 weeks. At the end of the study no statistically significant differences were noted between groups. However, significant improvements in both open comedones and total noninflammatory lesions were observed in the group washing twice a day. Worsening of acne condition was observed in the study group washing once a day, with significant increases in erythema, papules, and total inflammatory lesions. We concluded that slight support exists, both in terms of efficacy and convenience, for the recommendation to wash the face twice daily with a mild cleanser. However, excessive face washing may not be as culpable as previously thought.

Acne Vulgaris↗

Damage to the skin by repetitive washing.

Damage to the skin following the repeated use of 2 different wash solutions was investigated. Stratum corneum capacitative resistance, stratum corneum lipids, transepidermal water loss, skin surface pH, laser Doppler flow and skin reddening were determined. All skin function parameters already showed a marked change after a single wash (e.g., the median of TEWL values increased by more than 0.5 g/m2 h). Repetitive washing for 1 week led to a further deterioration of TEWL and corneometry values (e.g., TEWL increased for about 2.9 g/m2 h after repetitive washing with sodium lauryl sulfate). The rate of skin function regeneration after repetitive washing was unaltered compared to regeneration after a single wash. The surfactants used showed quantitatively differing effects on corneometry, TEWL and laser Doppler flow (e.g., after repetitive washing with Geliderm, the median of TEWL values increased only up to 1.55 g/m2 h). However, damage arising from repetitive washing could not be completely prevented by the selection of a mild surfactant. Skin function regeneration showed no difference with the 2 surfactants used.

Adult↗

Predictive washing test for evaluation of individual eczema risk.

It was the aim of our studies to estimate predictively the individual eczema risk for persons due to repetitive contact with washing-active substances, by a barrier function test on clinically healthy skin over 2 weeks. Within the scope of the study 3 groups with different atopy scores were compared. As washing solutions, 0.1 m SLS and a slightly acid soap-free washing emulsion were used in comparison to tap water. Prior to the 1st washing procedure, on days 3, 5, 8, 10, and 12, the transepidermal water loss, the horny layer moisture, and the skin blood flow were measured as parameters of barrier function, as well as the inflammatory reaction. The results prove that the atopy score has only limited validity as a predictive method for the acceptance of washing-active substances. The repetitive washing test, however, seems to be more adequate for evaluating the individual barrier function as well as the eczema risk. Irritation by a washing procedure may be greatly influenced by choice of the washing solution.

Adult↗

[Basic study of Beutin's washed sheep blood agar plate used for selective screening of verocytotoxin-producing/enterohemorrhagic Escherichia coli (VTEC/EHEC)].

Enterohemolysin production of Beutin's washed sheep blood agar plate has been used as an epidemiological marker for selective screening of Verocytotoxin-producing/Enterohemorrhagic Escherichia coli (VTEC/EHEC). In this study, we examined about the component of Beutin's washed sheep blood agar for further improvement of hemolysin production media. The following items were studied: the numbers of washings of sheep blood with phosphate buffered saline (PBS) (pH 7.2), concentration of sheep blood, kind and concentration of divalent metal ion (Ca2+, Mg2+) and basal medium. Twenty-seven strains of VT-producing E. coli of 7 different O-serotypes, 74 strains of VT-nonproducing E. coli of 24 different O-serotypes and one strain of O157 coded Escherichia hermanii were used for this basic study. In comparison of washing times of sheep blood with PBS, 5 times washing was better than 3 times, the original. In sheep blood concentration, supplement with 4% sheep blood was best for hemolysis observation. In experiment of addition of 2 divalent metal ions, Ca2+ and Mg2+, supplement with Ca2+ was more suitable than Mg2+ for hemolysis, and the supplement with 10 mM CaCl2, the original, was the best concentration. On the basal medium used in Beutin's sheep washed blood agar, 4 kinds of media were compared. In addition to Soybean-Casein Digest (SCD) agar, the original, Nutrient agar, Heart Infusion (HI) agar and Brain Heart Infusion (BHI) agar were examined, HI agar was the best blood agar base in the four media. As the above experimental results, the composition of the better Beutin's washed sheep blood agar may be summarize as follows: Heart Infusion agar 'Eiken' used for blood agar base, supplemented with 10 mM CaCl2 and 4% defibrinated sheep blood (Japan biosupp. Center) washed five times in phosphate-buffered saline, pH 7.2.

Animals↗

Role of receptor protein and membrane lipids in xanomeline wash-resistant binding to muscarinic M1 receptors.

Xanomeline is a novel agonist functionally selective for muscarinic receptors of the M1 subtype. It binds to this receptor in two modes, reversible and quasi-irreversible (wash-resistant). We investigated the unknown mechanism of the wash-resistant binding in experiments with muscarinic M1 receptors expressed in transfected Chinese hamster ovary cells. Xanomeline's structure consists of two heterocycles and O-hexyl side chain. We compared the wash-resistant binding of xanomeline and its analogs with shorter O-alkyl side chains. For the wash-resistant binding to occur, the O-alkyl chain had to be at least O-butyl or longer. Accumulation of inositol phosphates was enhanced in washed cells that had been preexposed to xanomeline or its pentyl analog, whereas the agonistic effects of the methyl, propyl, and butyl analogs were abolished by washing. Only the reversible binding of xanomeline was detected purified soluble receptors, but both binding modes occurred purified receptors reconstituted into liposomes and exposed xanomeline only after reconstitution. The wash-resistant binding did not occur if the exposure of purified receptors or liposomes alone to xanomeline, followed by washing, reconstitution. Simultaneous presence of receptors and lipid environment is therefore essential for the binding to take place. We suggest that the binding of xanomeline involves interhelical penetration of M1 muscarinic receptor by xanomeline's O-alkyl chain and interaction with membrane lipids surrounding the receptor.

Animals↗