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Adenine nucleotide content of corn roots as affected by injury and subsequent washing.

The adenine nucleotide content of the 2-centimeter segments excised from tray-grown corn (Zea mays L., WF9 x Mo17) roots declines for the first hour after excision. Concomitant with the loss of adenine nucleotides is a decline in respiration and a leakage of K(+). With continued washing, these parameters partially or completely recover and increased phosphate influx develops. Increasing the wound effect by cutting 0.5-centimeter segments gives a more rapid and pronounced degradation of adenine nucleotides and slower recovery. Conversely, the mild injury caused by submerging intact roots induces less degradation and produces greater net adenine nucleotide synthesis during recovery; adding auxin to the washing medium produces a similar result. With all treatments, there is stabilization of energy charge at about 0.85.Brief submersion or rubbing of intact roots, as well as recutting washed and recovered root segments, will initiate the transient loss of adenine nucleotides but will not induce increased phosphate influx.It is suggested that the loss in adenine nucleotides may reflect homeostasis in energy charge via catabolism arising from membrane permeability changes.

Journal Article↗

Markedly elevated levels of inflammatory cytokines in maternal serum and peritoneal washing during arrested labor.

BACKGROUND: The role of inflammatory cytokines in the pathophysiology of arrested labor is currently unknown. We measured levels of inflammatory cytokines in maternal serum and peritoneal washings in arrested first stage of labor to assess the possible involvement of these mediators in this obstetric disorder. METHODS: This was a prospective, case-control study involving 15 women who underwent cesarean section for arrested cervical dilatation (group I), 15 controls who were operated during active labor for nonreassuring fetal heart-rate tracing (group II) and 15 controls who were operated electively (group III). Blood samples were drawn from all women shortly before the operation. The presence of peritoneal fluid was assessed and peritoneal washings were obtained during the operation. All samples were assayed for the inflammatory cytokines interleukin (IL)-1beta, IL-6, IL-8 and soluble IL-2 receptor (sIL-2R) by solid-phase enzyme-linked immunosorbent assay (ELISA). RESULTS: All women from group I and one patient (6.7%) from group II had some degree of peritoneal fluid accumulation, while none from group III had any. Serum samples from group I contained significantly higher IL-1beta, IL-6, IL-8 and IL-2R levels than both control groups. Peritoneal washings from group I contained significantly higher IL-1beta, IL-6 and IL-8 but similar IL-2R levels. CONCLUSIONS: Arrested first stage of labor is associated with peritoneal fluid accumulation and increased levels of inflammatory cytokines in both serum and peritoneal fluids. Inflammatory cytokines may therefore be involved in the pathophysiology of arrested labor.

Adult↗

The effect of washing on the thickness of the stratum corneum in normal and atopic individuals.

The thickness of, and number of cell layers in, the stratum corneum and the living epidermis were determined on frozen sections of washed and unwashed skin from normal and atopic individuals of both sexes. The stratum corneum of atopic patients was thinner and had fewer layers of dead cells and intercellular lipid than normal, although the living epidermis was thicker. Regular washing with soap and water had no appreciable effect on the dimensions of the living epidermis of either group but caused a reduction in the number of cell layers and the amount of sudanophilic material in the stratum corneum of both. In atopic patients little surface lipid remained, suggesting that washing with soap and water may be detrimental to the barrier function of the stratum corneum in such patients.

Administration, Topical↗

The influence of daily dish-washing with synthetic detergent on human skin.

The effects of regular dish-washing on the stratum corneum barrier function, as determined by transepidermal water loss (TEWL), and objective and subjective skin parameters, were investigated in a 'use test' performed by 18 healthy volunteers. Hands were soaked in a 0.05% solution of a commercial dish-washing product (three times/day, for 15 min, at 37 degrees C) over a period of 3 weeks; one hand was unprotected and the other was protected with a commercial rubber glove. TEWL increased in 13/18 volunteers by more than 25% above baseline on exposed hands within the first 2 weeks of the study. Objective skin signs (erythema, scaling, fissures) and subjective symptoms (itching, dryness, smarting) occurred almost exclusively in subjects with substantial TEWL increases and were most prominent 1-2 weeks following peak TEWL values. There was a highly significant correlation between cumulative relative symptom scores and TEWL changes. TEWL increase and symptom scores were not correlated with a history of inhalant allergy and/or elevated serum IgE levels. Three volunteers, who had shown the highest increase of TEWL values and the most intense clinical reactions to the detergent were subjected to a control experiment in which one hand was soaked in warm tap-water following the same experimental protocol. No significant effects on TEWL values or skin symptoms were observed. We conclude that regular exposure to low concentrations of detergents as used for dish-washing is capable of inducing skin lesions in a substantial proportion of individuals.

Adult↗

Evaluation of the contribution of red and white cells to the flow of suspensions of washed blood cells through 3 micron Nuclepore membranes.

This study was designed to investigate a method of analysis which can quantitate the contribution of white cells to the flow of washed suspensions of blood cells. Such an analysis would obviate the need to remove white cells when studying the filterability of normal and abnormal red cells. The flow of suspensions of washed blood cells through a 3 micron Nuclepore membrane declines continually due to the occlusion of pores and the degree of pore occlusion is reduced significantly by the removal of the 'buffy coat' during the preparation of the suspension. These findings are in complete agreement with many other reports. However, a detailed kinetic analysis of the results suggests strongly that the white cell population makes a small contribution to the degree of pore occlusion which is caused largely by approximately 1% of the total red cell population. Despite the phenomenon and extent of pore occlusion, a kinetic analysis of the complete non-linear flow profile allows a measure of the deformability of red cells from filtration studies of washed but unfractionated blood cells.

Blood Flow Velocity↗

Exfoliated malignant cells in glove and instrument washings following head and neck surgery.

Loco-regional recurrence following resection of squamous cell carcinoma of the upper aero-digestive tract is a major problem. Cells exfoliated from malignant tumours have been considered capable of implanting and giving rise to local recurrence or metastatic deposits. A prospective study was performed examining washings from the gloves and instruments used in major head and neck operations to determine if exfoliated cells were indeed present. Following removal of the main tumour specimen all surgeons washed their hands in a solution containing a cell fixative and all instruments were soaked in the solution. Samples were then centrifuged and stained using a modified PAP technique. On occasions nuclear fragments consistent with squamous cell carcinoma were present in both glove and instrument washings. We recommend that all surgeons change gloves and instruments following removal of the main tumour specimen and prior to irrigation of the operative field with a tumoricidal agent.

Carcinoma, Squamous Cell↗

Intrauterine insemination of washed spermatozoa for treatment of oligozoospermia.

Efficacy of intrauterine insemination (IUI) using washed spermatozoa for treatment of oligozoospermia was evaluated by a prospective randomized study in 50 couples, using LH-timed natural intercourse in the alternate menstrual cycles as a control. The quality of spermatozoa in terms of their concentration and motility before and after sperm washing was compared. Sperm motility increased significantly after sperm preparation but the number of sperm was reduced. Eight pregnancies occurred in 253 cycles of IUI with washed spermatozoa and clomiphene citrate-stimulated cycles (3.16% per cycle). Only one patient conceived in 242 LH-timed natural intercourse cycles (0.41% per cycle). Compared with LH-timed natural intercourse, IUI provided a significantly improved pregnancy rate. When the sperm count was < 5 x 10(6) per ml, no pregnancy occurred with the IUI method. Therefore, IUI is a of rather limited usefulness when the sperm quality is very poor. Few complications occurred after IUI, but included slight cervical contact bleeding and mild abdominal discomfort and/or cramps. In conclusion, IUI should be considered as a useful and relatively non-invasive therapeutic modality for treating caused by moderate oligozoospermia (> 5 x 10(6)/ml), when sexual intercourse fails.

Clomiphene↗

Investigations of the effectiveness of detergent washing, drying and chemical disinfection on contamination of cleaning cloths.

Detergent washing, drying and chemical disinfection for decontamination of cleaning cloths was investigated with cloths contaminated by use in the domestic environment. Detergent washing produced only limited reductions in microbial contamination and cloths then stored at room temperature for 24 h showed increases in contamination due to multiplication of residual survivors. For effective and consistent decontamination of cloths, detergent washing followed by drying at 80 degrees C for 2 h was required. Hypochlorite and phenolic disinfectants produced significant reductions in contamination, but chemical disinfection may be unreliable where cloths are heavily contaminated.

Bacteria↗

The influence of the time of etching and washing on the bond strength of fissure sealants applied to enamel.

Variation of the etching and washing times of enamel surfaces prior to the application of fissure sealants affect the bond strength in differing ways. Depending on the length of the etch time, the effect of washing may increase or decrease the bond strength of the sealant. Clinically, the most satisfactory combination of etch and wash time was found to be different to that suggested by the manufacturers.

Acid Etching, Dental↗

Dentine hypersensitivity: uptake of toothpastes onto dentine and effects of brushing, washing and dietary acid--SEM in vitro study.

Toothpastes are commonly used in the treatment of dentine hypersensitivity. One mode of action could be to block patent dentinal tubules with toothpaste ingredients or by abrasion to form a smear layer. Studies in vitro have shown that toothpaste abrasives have variable affinity for the dentine surface but retention may be limited. This SEM study extended previous work and determined the uptake in vitro of toothpaste ingredients onto the dentine surface and their retention when challenged by washing with water or orange juice. All products were seen to leave variable residues on the surface of etched or unetched dentine specimens, furthermore brush application produced some smearing with closure of previously open tubules. Washing with water and more particularly orange juice removed most residues and orange juice etched the dentine to open previously closed tubules. One product with an artificial silica abrasive was relatively resistant to both water and orange juice washings and in a pilot experiment showed some penetration into tubules not seen with another desensitizing product. These findings suggest that some abrasives may be usefully employed in desensitizing products, but without consideration of erosive factors in the aetiology of dentine hypersensitivity recurrences of the condition will be common.

Acid Etching, Dental↗

Leucocyte-associated plasma proteins. I. Plasma proteins in supernatants and cell pellets during successive washings of human blood cells.

Sixteen plasma proteins were studied in the supernatants from 20 successive washings of human leucocytes by a new sensitive immunoelectrophoretic method. In the first few supernatants a fast and parallel decrease in concentration of all proteins took place, but for 11 proteins this was followed by a phase of slow decrease due to cellular release. Eight proteins could be quantitated throughout the 20 washings: prealbumin, albumin, orosomucoid, alpha 1-antitrypsin, transferrin, C3, haptoglobin, and IgG. These proteins and trace amounts of eight other plasma proteins were present in the cell lysate after the washings. In comparison with plasma, the leucocytes contained relatively higher concentrations of prealbumin, orosomucoid, alpha 1-antitrypsin, and haptoglobin than of albumin. In crossed immunoelectrophoresis these same four proteins were present as molecular variants differing from their plasma counterpart in electrophoretic mobility and morphology of the precipitate.

Animals↗

Spontaneous histamine release from washed leukocytes and whole blood in atopic and non-atopic individuals.

The ability of basophils from patients with allergic rhinitis, extrinsic asthma, chronic idiopathic urticaria and atopic dermatitis to release histamine spontaneously in vitro was studied. Spontaneous release in the presence and absence of 30% D2O was investigated from both washed leukocytes and whole blood. Compared with controls histamine release (HR) from washed leukocytes was significantly enhanced in allergic rhinitis patients only, whereas in the other groups only a certain percentage of patients was found to have high spontaneous HR. In whole blood experiments HR in all groups was within the normal range. Our data indicate that enhanced spontaneous mediator release from washed basophils in vitro does not necessarily prove this mechanism to be of pathophysiological relevance in vivo.

Adolescent↗

Preparation of leukocyte-poor blood: a comparison of IBM 2991 washing and Huggins freeze-thawing.

Frequently, forzen blood is requested in order to provide a leukocyte-poor red cell transfusion and a reduced level of exposure to hepatitis virus rather than in an effort to find serological compatible red blood cells. However, this usage of frozen blood is expensive and, as recently reported, does not eliminate the risk of transmission of hepatitis. To assess the feasibility of substituting washed cell preparations, we have compared the Huggins frozen blood process with the IBM 2991 cell washing technique and have evaluated the efficiency of leukocyte removal and red cell recovery from homologous pools of fresh blood. The results of these experiments combined with the greatly decreased cost (approximately 1/3) and time required (approximately 1/6) indicate that many of the requests which blood banks receive for frozen blood could, in fact, be better met by using cells which have been washed in the IBM 2991.

Blood Preservation↗

A new approach to washing red blood cells frozen with a high concentration of glycerol in a special freezing container.

We report here on a new approach to washing red blood cells frozen with a high concentration of glycerol in a special freezing container. The wash solution consists of a 150-ml volume of 12% sodium chloride and 2 liters of 0.9% sodium chloride-0.2% glucose-25 mEq/l disodium phosphate. Both the Haemonetics Blood Processor 115 and the IBM Blood Processor 2991 have been used with this protocol, with similar results. The in vitro recovery of red blood cells frozen with 8.6M glycerol was 89%, and that of red blood cells frozen with 6.2M glycerol was 93%. The 24-hour posttransfusion survival values averaged 88% for eight units of outdated-rejuvenated previously frozen red blood cells washed by this protocol and stored at 4 degrees C for 3 days before autotransfusion.

Blood Preservation↗

Manipulating red cell intra- and extracellular pH by washing.

Washing red cells with solutions containing no anions capable of entering the cells is known to result in the loss of intracellular chloride and a counterflow of OH- which raises intracellular and lowers extracellular pH. Elevating the intracellular pH improves the quality of the cells during 4 degrees C storage. The extent to which intracellular pH can be raised and extracellular pH reduced depends not only on the inability of extracellular anions to enter the cells but also on the buffering capacity of the wash solution. Both intra- and extracellular pH can be manipulated by the judicious selection of anions or combinations of anions used in the wash solution.

Blood Preservation↗

Steady-state pharmacokinetics of oestradiol gel in post-menopausal women: effects of application area and washing.

OBJECTIVE: To investigate the effect of cutaneous application area on oestradiol absorption using an oestradiol gel for transdermal use. Furthermore, the effect of washing of the application site on oestradiol pharmacokinetics was studied. DESIGN: Open-label, randomised, three-way cross-over study. SETTING: A clinical pharmacokinetic research unit. SUBJECTS: Sixteen healthy postmenopausal volunteers. INTERVENTIONS: During three treatment periods, subjects were treated with 1 mg oestradiol (1.0 g Divigel/Sandrena 0.1% gel), applied on the thigh to a skin area of 200 cm2, 400 cm2, or on an area 'as large as possible'. Blood samples were drawn during steady-state (on days 14-15) immediately before application and at regular time intervals thereafter. MAIN OUTCOME MEASURES: Serum oestradiol levels were measured for determination of peak concentration (Cmax), time to peak concentration (tmax) and bioavailability using the area under the time concentration curve (AUC0-24). RESULTS: Bioavailability from a 200 cm2 area was 2-fold higher than that from the largest possible area (P < 0.05). Oestradiol peak plasma concentration from a 200 cm2 area was higher than from the two larger areas (P < 0.05), which did not differ significantly from each other. Washing of the application site 30 min after application reduced the bioavailability from both the 200 cm2 and 400 cm2 application areas (P < 0.01), but peak plasma concentration from the 200 cm2 area only (P < 0.01). Time to reach peak plasma concentrations was significantly reduced with all three application areas by washing. CONCLUSION: A higher oestradiol absorption was achieved from a smaller application area. No marked differences were observed in the pharmacokinetics between 200 cm2 and 400 cm2 application areas, which are those recommended for this oestradiol gel preparation.

Administration, Cutaneous↗

Benzodiazepine receptor binding following amygdala-kindled convulsions: differing results in washed and unwashed brain membranes.

The binding of [3H]flunitrazepam and [3H]RO5-4864 was measured in unwashed brain homogenates and in extensively washed brain membranes from amygdala-kindled and "yoked" control rats sacrificed 2 weeks following the sixth stage 5 convulsion. In unwashed homogenates, [3H]flunitrazepam binding was reduced in both the hypothalamus and ipsilateral right cortex of kindled rats (unchanged in other areas). In washed brain membranes, [3H]flunitrazepam binding was unaltered in these regions; it was bilaterally elevated, however, in both the amygdala and hippocampus (unchanged in other areas). In washed membranes, the in vitro addition of gamma-aminobutyric acid enhanced [3H]flunitrazepam binding to a similar extent in kindled and control membranes. These data indicate that the type of benzodiazepine binding abnormality observed after kindling depends on the type of tissue preparation employed in the assay procedure.

Amygdala↗

Single and multiple breath nitrogen wash-out--closing volume and volume of trapped gas for detection of early airway obstruction.

Thirty-eight patients with varying degrees of peripheral airway involvement were studied by flow-volume curves and two nitrogen wash-out techniques in an on-site study. Closing capacity (CC), closing volume in percentage of vital capacity (CV%), slope of the alveolar plateau (phase III), volume of trapped gas (VTG), and maximum expiratory flow at 50 and 75% of vital capacity (MEF50, MEF25) were determined and correlation analyses were performed. A biphasic or exponential correlation was noted between the degree of airway closure and VTG with rapidly increasing VTG at high CC. Although there were correlations between spirometry and the two nitrogen wash-out techniques, a large scatter precluded the use of the flow-volume variables to predict CC, CV% and VTG. The coefficient of variation was lower for the measurements of CV% than for the recording of VTG. Since the single breath nitrogen wash-out technique is also technically simpler it is recommended prior to the multiple breath technique.

Adult↗