Wash Period Research Articles

Neuronal nicotinic acetylcholine receptors are modulated by zinc

It is known that zinc modulates nicotinic acetylcholine receptors (nAChRs). Here, we studied the effects of zinc on neuronal α4β4 nAChRs, expressed in Xenopus oocytes and activated by nicotine. Membrane ion currents elicited by nicotine (10 nM to 100 μM) were enhanced by zinc (100 μM). Maximal zinc potentiation of the nicotine-activated current (2530%) occurred at 50 nM nicotine, and potentiation gradually decreased as the nicotine concentration increased. The EC 50 and IC 50 for the nicotine-activated current were 639 nM and 14.7 μM nicotine, respectively. Both parameters decreased in the presence of zinc to 160 nM and 4.6 μM, respectively, probably due to an increase of sensitivity of nAChRs for nicotine. We used different concentrations and durations of exposure to nicotine, due to desensitization of nAChRs directly depends on both these factors. With 500 nM nicotine and 20 min washing periods between nicotine applications, zinc potentiation remained constant, 901% for 2 min and 813% for 20 min of nicotine exposure. With continuous application of nicotine, zinc potentiation decreased as the time of nicotine exposure increased, 721% for 2 min and 254% for 48 min of nicotine exposure. Our results indicate that zinc-potentiating effects on α4β4 nAChRs strongly depend on both concentration and time of exposure to nicotine, suggesting that zinc potentiation depends on the degree of desensitization.

Endothelin-1 Inhibits Thick Ascending Limb Transport via Akt-stimulated Nitric Oxide Production

Endothelin-1 inhibits sodium reabsorption in the thick ascending limb (THAL) via stimulation of nitric oxide (NO) production. The mechanism whereby endothelin-1 stimulates THAL NO is unknown. We hypothesized that endothelin-1 stimulates THAL NO production by activating phosphatidylinositol 3-kinase (PI3K), stimulating Akt activity, and phosphorylating NOS3 at Ser1177. This enhances NO production and inhibits sodium transport. We measured 1) NO production by fluorescence microscopy using DAF2-DA, 2) Akt activity using a fluorescence resonance energy transfer-based Akt reporter, 3) phosphorylated NOS3 and Akt by Western blotting, and 4) NKCC2 activity by fluorescence microscopy. In isolated THAL, endothelin-1 (1 nmol/liter) increased NO production from 0.23 +/- 0.24 to 2.81 +/- 0.32 fluorescence units/min (p < 0.001; n = 5) but failed to stimulate NO production in THALs isolated from NOS3-/- mice. Wortmannin (150 nmol/liter), a PI3K inhibitor, reduced endothelin-1-stimulated NO by 83% (0.49 +/- 0.13 versus 3.31 +/- 0.49 fluorescence units/min for endothelin-1 alone; p < 0.006; n = 5). Endothelin-1 stimulated Akt activity by 0.16 +/- 0.02 arbitrary units as measured by fluorescence resonance energy transfer (p < 0.001; n = 5) and increased phosphorylation of Akt at Ser473 by 56 +/- 11% (p < 0.002; n = 7). Dominant-negative Akt blocked endothelin-1-induced NO by 60 +/- 8% (p < 0.001 versus control; n = 6), and an Akt inhibitor had a similar effect. Endothelin-1 increased phosphorylation of NOS3 at Ser1177 by 89 +/- 24% (p < 0.01; n = 7) but had no effect on Ser633. Endothelin-1 inhibited NKCC2 activity, an effect that was blocked by dominant-negative Akt and NOS inhibition. We conclude that endothelin-1 stimulates THAL NO production by activating PI3K, stimulating Akt activity, and phosphorylating NOS3 at Ser1177. This enhances NO production and inhibits sodium transport.

Cortisol and prolactin modulation of caudal neurosecretory system activity in the euryhaline flounder Platichthys flesus

Previous studies have shown roles for cortisol and prolactin in osmoregulatory adaptation to seawater and freshwater, respectively, in euryhaline fish. This study of the European flounder investigated the potential for these hormones to modulate activity of the caudal neurosecretory system (CNSS), which is thought to be involved in physiological adaptation to changing external salinity. Superfusion of isolated CNSS with either cortisol or prolactin (10 μM; 15 min) led to changes in firing activity in neuroendocrine Dahlgren cells, recorded extracellularly. Cortisol evoked a modest increase in overall firing activity, with the response delayed by 4 h after treatment. The response to prolactin was short latency, continued to build up over the subsequent 4-h wash period, and comprised increased firing activity together with recruitment of previously silent Dahlgren cells. Immunoreactivity for glucocorticoid and prolactin receptors was localised to Dahlgren cells. The CNSS expression level for glucocorticoid-2 receptor mRNA, measured by Q-PCR, was significantly lower in fish fully acclimated to freshwater, compared to seawater. No differences were seen between these two states for prolactin receptor mRNA expression. These results provide evidence for a modulatory action of both hormones on the neurosecretory function of the CNSS.

Effect of pre-freezing conditions on semen cryopreservation in rhesus monkeys

Although sperm cryopreservation has been studied in at least 17 non-human primate species, systematic factor optimization for any single species is lacking. Gene banking of non-human primate sperm is still in its infancy. The objective of the present study was to initiate a systematic approach to optimize the process of sperm cryopreservation for rhesus macaques, specifically, factors related to pre-freezing conditions (e.g., straw freezing position, sperm concentration, sperm washing, equilibration methods, and equilibration time periods). Straw position had no effect on post-thaw motility (P=0.193). Sperm concentration was tested in a range from 5 x 10(6)mL(-1) to 5 x 10(8)mL(-1); post-thaw motility of sperm samples frozen at 5 x 10(7)cell mL(-1) (51.0+/-10.6%; mean+/-S.D.) and 5 x 10(8)cell mL(-1) (48.1+/-7.3%) were higher than samples frozen at 5 x 10(6)cells mL(-1) (33.0+/-12.0%, P=0.003). Comparison of motility immediately after thawing between samples with (51.2+/-6.2%) and without washing (53.9+/-6.8%) revealed no differences (P>0.05). However, washing improved sperm forward progression within 1h after thawing, whereas unwashed sperm retained higher post-thaw motility and progression during extended incubation (4h) after thawing (P<0.05). Equilibration methods (with or without pre-cooling) made no difference on post-thaw motility (P>0.05), and the most effective equilibration time was the duration required for samples to acclimate to 4 degrees C prior to freezing. Evaluation and optimization of these pre-freezing conditions will help to minimize sources of injury, maximize survival, and contribute to the development of an optimized cryopreservation protocol for rhesus macaque sperm.

An improved protocol for Agrobacterium-mediated transformation of grapevine (Vitis vinifera L.)

An improved protocol for efficient Agrobacterium-mediated transformation of grapevine (Vitis sp.) was developed through modification of cocultivation and subsequent washing procedures. It was determined that Agrobacterium-infected somatic embryos (SE) cocultivated on filter paper exhibited less browning and significantly higher transient GFP and GUS expression than those cultured on agar-solidified medium. Furthermore, such SE, when subjected to a prolonged washing period in liquid medium containing cefotaxime and carbenicillin, followed by another wash in similar medium with kanamycin added, exhibited significantly higher rates of stable transformation compared to previously-described procedures. Transgenic plant recovery was increased 3.5–6 Xs by careful excision of leafy cotyledons from SE that had been induced to germinate on MS medium containing 1 μM of BA. Southern blot analysis revealed the low copy number integration of transgenes in transgenic plants recovered using the improved protocol. These improved cocultivation and plant recovery procedures have been demonstrated to facilitate production of large populations of transgenic plants from V. vinifera ‘Merlot’, ‘Shiraz’ and ‘Thompson Seedless’ as well as Vitis hybrid ‘Seyval Blanc’.

Cytidine‐5′‐diphosphocholine (Citicoline): a pilot study in patients with non‐arteritic ischaemic optic neuropathy

Our work evaluates visual function before and after treatment with cytidine-5-diphosphocholine (Citicoline) in patients with non-arteritic ischaemic optic neuropathy (NION). Twenty-six patients in which at least 6 months elapsed from NION, were randomly divided into two age-similar groups: 14 patients had Citicoline (Cebrolux-Tubilux, Italy, 1600 mg/diem for 60 days, followed by a 120-day period of wash out, days 60-180) (T-NION); 12 patients had no treatment during the same period (NT-NION). At day 180, in T-NION a second period of treatment (days 181-240) followed by a wash-out (days 241-360) was performed. Fourteen age-matched healthy subjects provided normative data. In all patients, pattern-electroretinogram (PERG), visual evoked potentials (VEPs) and visual acuity (VA) measurements were performed at baseline and at days 60 and 180. In T-NION, further measurements were achieved at days 240 and 360. At baseline, NT-NION and T-NION patients showed abnormal PERGs and VEPs, and reduced VA, compared to controls. At the end of treatment (days 60 and 240), T-NION patients showed improvement (P < 0.01) of PERGs, VEPs parameters and VA, compared to pre-treatment values. After wash out, functional improvements persisted compared to baseline. No changes in NT-NION patients were observed. Our results suggest a beneficial effect of oral Citicoline in NION.

The Impact of Design Variables and Aftercare Regime on the Long-Term Performance of Pressure Garments

We sought to establish the impact of pressure garment design variables, moisturizer use, and laundry method on the ability of pressure garments to maintain their pressure delivering potential, indicated here by garment tension, over time and use. Twenty-six sets of three replicate pressure garment sleeves were constructed from four powernet fabrics, using three reduction factors and six sleeve dimensions. These pressure garment sleeves were extended for 23 hours on static cylinder models followed by hand or machine laundry up to 28 times. Some sleeves were additionally exposed to moisturizers during their extension. Garment tension and dimensions were measured before and during the simulated wear and wash period to indicate each garment's ability to maintain its tension and therefore pressure throughout a period of "use." The results of the investigation were analyzed in groups where each group contained only 1 variable, thereby allowing the variables with the most significant impact on tension degradation to be identified. The investigation confirmed that all pressure garments lost tension and therefore pressure delivering ability over time and use. It further revealed that pressure garments designed to exert greater pressures degraded faster than those designed to exert lower pressures. Contact between pressure garments and moisturizers accelerated tension degradation, and machine-washing pressure garments tended to prolong their pressure-delivering properties compared with hand-washing them. To maintain the initial pressure delivered by pressure garments, powernet fabrics should be prestressed before being designed/constructed and they should be machine-washed by patients.

Preparation of Zirconia Nanopowders in Ultrasonic Field by the Sol-Gel Method

Zirconia nanopowders were prepared in the ultrasonic field by the sol-gel method and the sonochemical effect on the structure of zirconium hydroxide and the zirconia nanopowder properties were systematically investigated in this work. Ultrasound was introduced into the different stages of the synthesis of zirconia nanopowders in sol-gel reaction system, and zirconium hydroxides and the zirconia nanopowders with different properties were obtained. The results indicated that ultrasonic cavitation could not only disaggregate the agglomerates of zirconia colloidal particles but also reduce the amount of coordinated H2O, free H2O and free hydroxyl groups of the zirconium hydroxide colloidal particles, thus effectively preventing the formation of hard agglomerates in zirconia powders. Moreover, the effects of different ultrasonic output powers and treatment cycles on the structure and properties of ZrO2 nanopowders were studied by TEM, XRD and SAXS. Zirconia nanopowders with an extremely small crystallite size (10.3 nm) and a narrow size distribution were yielded with 520 W ultrasound for 6 treatment cycles on the formation period and 600 W ultrasound for 2 treatment cycles on the washing period. It is concluded that the ultrasonic field is a potential method for nanopowder preparation.

Nitric oxide modulates angiotensin II-induced endothelial vasoconstrictor prostanoid release

This study investigated the modulation of angiotensin II-induced endothelial prostanoid release in rabbit aortic rings. Two cumulative dose response curves with 90-min washing interval were performed. Incubation with l-NG-nitroarginine methyl ester (l-NAME) 10−4 M increased angiotensin II maximal contractile response (Emax). This effect was reversed by indomethacin 10−5 M, diphenyliodinum 10−5 M, Tempol 10−5 M or ascorbic acid 10−4 M in both cumulative dose response curves and by SQ 29548 10−6 M in the second cumulative dose response curve. When segments were treated with tetraethylamonium 10−3 M but not with glibenclamide 10−5 M during the washing period, l-NAME recovered its ability to enhance the Emax in arteries incubated with SQ 29548. Conclusions: nitric oxide modulates angiotensin II-induced endothelial release of cyclooxygenase-dependent eicosanoids, one of which acts through thromboxane A2/prostaglandin H2 receptors and would decrease KCa channel activity. An increase in free radical production may account for the enhancement of such prostanoid release. Furthermore, it was found that in the present conditions, the release of the hyperpolarizing factor would improve in order to maintain the vascular tone.

Autoregulatory Role of Endothelium-derived Nitric Oxide (NO) on Lipopolysaccharide-induced Vascular Inducible NO Synthase Expression and Function

Nitric oxide (NO) produced by inducible nitric oxide synthase (iNOS) is responsible for sepsis-induced hypotension and plays a major contributory role in the ensuing multiorgan failure. The present study aimed to elucidate the role of endothelial NO in lipopolysaccharide (LPS)-induced iNOS expression, in isolated rat aortic rings. Exposure to LPS (1 mug/ml, 5 h) resulted in a reversal of phenylephrine precontracted tone in aortic rings (70.7 +/- 3.2%). This relaxation was associated with iNOS expression and NF-kappaB activation. Positive immunoreactivity for iNOS protein was localized in medial and adventitial layers of LPS-treated aortic rings. Removal of the endothelium rendered aortic rings resistant to LPS-induced relaxation (8.9 +/- 4.5%). Western blotting of these rings demonstrated an absence of iNOS expression. However, treatment of endothelium-denuded rings with the NO donor, diethylamine-NONOate (0.1 mum), restored LPS-induced relaxation (61.6 +/- 6.6%) and iNOS expression to levels comparable with arteries with intact endothelium. Blockade of endothelial NOS (eNOS) activation using geldanamycin and radicicol, inhibitors of heat shock protein 90, in endothelium-intact arteries suppressed both LPS-induced relaxation and LPS-induced iNOS expression (9.0 +/- 8.0% and 2.0 +/- 6.2%, respectively). Moreover, LPS treatment (12.5 mg/kg, intravenous, 15 h) of wild-type mice resulted in profound elevation of plasma [NO(x)] measurements that were reduced by approximately 50% in eNOS knock-out animals. Furthermore, LPS-induced changes in vascular reactivity and iNOS expression evident in wild-type tissues were profoundly suppressed in tissues taken from eNOS knockout animals. Together, these data suggest that eNOS-derived NO, in part via activation of NF-kappaB, regulates iNOS-induction by LPS. This study provides the first demonstration of a proinflammatory role of vascular eNOS in sepsis.

Beta-adrenoceptor subtype dependence of chronotropy in mouse embryonic stem cell-derived cardiomyocytes.

Cardiomyocytes derived from embryonic stem cells (ESCM) have potential both as an experimental model for investigating cardiac physiology and as a source for tissue repair. For both reasons it is important to characterise the responses of these cells, and one of the key modulators of contraction is the beta-adrenergic system. We therefore undertook a detailed study of the response of the spontaneous beating rate of ESCM to beta-adrenoceptor (betaAR) stimulation. Embryoid bodies (EBs) were generated from murine ES line E14Tg2a by the hanging drop method, followed by plating. Spontaneously beating areas were seen starting from 9-14 days after differentiation: the experiments described here were performed on EBs between developmental day 19 and 48. Beating cell layers were seeded with charcoal to allow tracking of movement by a video-edge detection system. Experiments were performed in physiological medium containing 1 mM Ca2+ at 37 degrees C. Isoprenaline (Iso) increased beating rate with an EC50 value of 52 nM. Iso (0.3 microM) increased basal rate from 67 +/- 7 beats per minute (bpm) to 138 +/- 18 bpm, P < 0.001, n = 22. At earlier developmental time points the response to Iso was not maintained through 5 min exposure; this spontaneous desensitisation only being observed before day 36. A repeat application of Iso after a wash period of 20 min produced reproducible effects on beating rate. Subtype dependence of the betaAR response was determined by comparing an initial response with a second in the presence of selective beta1- or beta2AR antagonists. In the presence of the specific beta1AR-blocker CGP 20712A (300 nM) the increase in rate with Iso was reduced from 207 +/- 42% of basal to 128 +/- 13%, P < 0.01. With the beta2AR-blocker ICI 118,551 (50 nM) there was no significant change in Iso response. Exposure to the muscarinic agonist, carbachol (10 microM), inhibited the increase in frequency mediated by isoprenaline, but had mixed stimulatory and inhibitory effects on basal rate. This study extends the characterisation of ESCM as a preparation for studying receptor pharmacology, and indicates that the beta1AR is the predominant subtype mediating increases in contraction rate in murine ESCM.

Cleansing without compromise: the impact of cleansers on the skin barrier and the technology of mild cleansing.

Cleanser technology has come a long way from merely cleansing to providing mildness and moisturizing benefits as well. It is known that harsh surfactants in cleansers can cause damage to skin proteins and lipids, leading to after-wash tightness, dryness, barrier damage, irritation, and even itch. In order for cleansers to provide skin-care benefits, they first must minimize surfactant damage to skin proteins and lipids. Secondly, they must deposit and deliver beneficial agents such as occlusives, skin lipids, and humectants under wash conditions to improve skin hydration, as well as mechanical and visual properties. While all surfactants tend to interact to some degree with lipids, their interaction with proteins can vary significantly, depending upon the nature of their functional head group. In vitro, ex vivo, and in vivo studies have shown that surfactants that cause significant skin irritation interact strongly with skin proteins. Based on this understanding, several surfactants and surfactant mixtures have been identified as "less irritating" mild surfactants because of their diminished interactions with skin proteins. Surfactants that interact minimally with both skin lipids and proteins are especially mild. Another factor that can aggravate surfactant-induced dryness and irritation is the pH of the cleanser. The present authors' recent studies demonstrate that high pH (pH 10) solutions, even in the absence of surfactants, can increase stratum corneum (SC) swelling and alter lipid rigidity, thereby suggesting that cleansers with neutral or acidic pH, close to SC-normal pH 5.5, may be potentially less damaging to the skin. Mildness enhancers and moisturizing agents such as lipids, occlusives, and humectants minimize damaging interactions between surfactants, and skin proteins and lipids, and thereby, reduce skin damage. In addition, these agents play an ameliorative role, replenishing the skin lipids lost during the wash period. The present review discusses the benefits of such agents and their respective roles in improving the overall health of the skin barrier.

Olfactory signal modulation by molluscan cardioexcitatory tetrapeptide (FMRFamide) in axolotls (Ambystoma mexicanum).

The terminal nerve, which innervates the nasal epithelia of most jawed vertebrates, is believed to release neuropeptides that modulate activity of sensory receptor neurons. The terminal nerve usually contains gonadotropin-releasing hormone as well as at least one other peptide that has not been characterized, but which bears some structural similarity to molluscan cardioexcitatory tetrapeptide (FMRFamide) and neuropeptide tyrosine (NPY). We investigated the effects of FMRFamide on both voltage-gated currents and odorant responses in the olfactory epithelium of axolotls (Ambystoma mexicanum), using whole-cell patch clamp and electro-olfactogram (EOG) recording techniques. In the presence of FMRFamide, the magnitude of a voltage-gated inward current was dramatically increased, reaching an average of 136% of the initial (pre-exposure) magnitude in neurons that showed a response to the peptide. This increase is detectable within approximately 1-2 min of exposure to FMRFamide and is sustained for at least 10 min. In EOG experiments, odorant responses are not affected during FMRFamide application, but are sometimes increased or decreased during the subsequent wash period. On average, the largest single EOG response in each trial was detected approximately 25 min after initial FMRFamide application, and ranged from 110 to 147% of baseline. These results suggest that a compound similar to FMRFamide, if released from the terminal nerve, may function in peripheral olfactory signal modulation.

Gonadotropin releasing hormone (GnRH) modulates odorant responses in the peripheral olfactory system of axolotls.

Peripheral signal modulation plays an important role in sensory processing. Activity in the vertebrate olfactory epithelium may be modulated by peptides released from the terminal nerve, such as gonadotropin releasing hormone (GnRH). Here, we demonstrate that GnRH modulates odorant responses in aquatic salamanders (axolotls, Ambystoma mexicanum). We recorded electrical field potentials (electro-olfactograms, or EOGs) in response to stimulation with four different amino acid odorants, L-lysine, L-methionine, L-cysteine, and L-glutamic acid. EOG responses were recorded from the main olfactory epithelium before, during, and after application of 10 microM GnRH. This protocol was repeated for a total of three trials with 60-80 min between trials. The effect of GnRH on EOG responses was broadly similar across odorants and across trials. In general, EOG responses were reduced to 79% of the initial magnitude during application of GnRH; in some trials in which glutamic acid served as the odorant, EOG responses were enhanced during the wash period. Although the 4-min inter-stimulus interval did not lead to adaptation of EOG responses during the first trial, we frequently observed evidence of adaptation during the second and third trials. In addition, we found that lower concentrations of GnRH produced a smaller effect. These results demonstrate that GnRH can modulate odorant responses in the peripheral olfactory system.

THE RATE OF WASHING OF PAPER

Conservation washing of paper involves the removal of water-soluble materials. In order to study the factors which influence the rate and extent of washing, a model discoloured paper was made in the laboratory by immersing filter paper in a concentrated extract from old mounting board. The washing kinetics of this paper were studied by subjecting samples to various treatments performed for different times, and measuring the visible reflectance of samples. It was discovered that the treated paper became more difficult to wash as it aged and that a residue of the stain remained in the paper even when washed immediately after treatment. This observation may help to explain why stains such as ‘tide-marks’ are difficult to remove. It was demonstrated that increasing the temperature increased both the rate and the extent of washing. Seven different washing processes used in paper conservation were studied to determine the water content of the paper during the processes, and the rate and extent of washing at intervals during a two-hour washing period. Ultraviolet fluorescence revealed the uniformity of washing. The implications for conservation are discussed.

Desensitization of diliganded mouse muscle nicotinic acetylcholine receptor channels.

Nicotinic ACh receptor channels (AChRs) exposed to high concentrations of ACh adopt 'desensitized' conformations that have a high affinity for the transmitter and no measurable ion conductance. Single-channel currents elicited by 0.1 or 1 mM ACh were recorded from human embryonic kidney (HEK) cells that had been transiently transfected with mouse alpha, beta, delta, and epsilon subunits. On the time scale of approximately 0.1 ms to approximately 1 h, apparent open intervals are described by a single exponential component, and shut intervals associated with desensitization are described by the sum of four or five exponential components. The kinetic behaviour appeared to be stationary and homogeneous. Desensitization rate constants were estimated by kinetic modelling of currents from cell-attached and outside-out patches (where the number of channels in the patch was measured). A single AChR recovered from the longest-lived desensitized state only after approximately 5 min. The occupancy of an AChR for each of the desensitized states was calculated as a function of time after the continuous application of a pulse of saturating ACh. The longest-lived desensitized state accounted for 90 % of the total only after several seconds. The fractional recovery from desensitization (during a 200 ms wash period) decreased as the duration of the desensitizing pulse increased, suggesting that recovery is slower from the longer-lived desensitized states. The free energy landscape for the AChR desensitization reaction in cell-attached patches exhibited an initial destabilization, followed by a plateau region of gradually increasing stability, followed by a deep well.

The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat

This work was carried out to adapt the electrolyte leakage technique to durum wheat and then to evaluate its relevance in the assessment of the cell membrane stability as a mechanism of water stress tolerance in this species. The method currently used is based on in vitro desiccation of leaf tissues by a solution of polyethylene glycol (PEG) and a subsequent measurement of electrolyte leakage into deionised water. It consists of three successive steps: (1) a washing treatment to remove solutes from both leaf surfaces and cells damaged by cutting; (2) a stress period during which the leaf tissues are plunged in a PEG-solution and (3) a rehydration period during which after-effects of the stress are evaluated. During the washing period, the major part of electrolytes was removed within 15 min. Varying the stress conditions influenced both the percent and the kinetics of electrolyte leakage during rehydration. Electrolyte leakage exhibited a characteristic pattern reflecting the condition of cellular membranes (repair and hardening). In practice, we recommend a 15-minute washing time, a 10-hour stress period and 4 h of rehydration. The extent of the cell membrane damage not only correlated well with the growth responses of wheat seedlings belonging to various cultivars to withholding water but also with the recognised field performances of these cultivars. The relative proportion of endogenous ions lost in the effusate during the rehydration step may vary strongly according to the element analysed and the precise nutritional status of the plant should therefore be considered. However, an increase in inorganic ion leakage does not fully explain the recorded PEG-induced increase in electrical conductivity (EC) during the subsequent rehydration step and organic ions are probably also involved in such an increase.

Influence of maize starch granule-associated protein on the rheological properties of starch pastes. Part II. Dynamic measurements of viscoelastic properties of starch pastes

The influence of granule-bound starch synthase (GBSS) on viscoelastic properties of gelatinized starch pastes was studied using a normal, a waxy null, and two GBSS-containing waxy mutant maize starches (waxy protein was synthesized with no GBSS activity). The four starches were isolated using the toluene method (isolation procedure I) (IP I) and were then further purified using an extended period of washing (isolation procedure II) (IP II). Dynamic rheological measurements of IP I gelatinized starches showed that storage moduli ( G′) were higher in the two GBSS-containing waxy starches than in the waxy null mutant starch not containing GBSS. The total non-GBSS granule-associated proteins of the three waxy starches were similar. Further removal of granule-associated proteins by purification decreased the G′, with decreases higher in the two GBSS-containing waxy mutants than in the waxy null mutant starch. This was interpreted to be due to the removal of GBSS during purification of the former. High shear broke gelatinized IP I waxy and normal maize starches starch granule or remnant structure, which caused a decrease in G′. Cox–Merz plots showed that the elastic properties of the starch pastes were reduced by removal of starch granule-associated proteins.

Der repetitive Waschtest als Modell zur Beurteilung von Hautschutzpräparaten am Beispiel einer dexpanthenolhaltigen Formulierung

Der repetitive Waschtest mit NLS stellt ein Irritationsmodell dar, mit dem Hautschutzpräparate auf ihre Wirksamkeit gegenüber hydrophilen Reizstoffen überprüft werden können. An eine repetitive Waschphase mit vorheriger Anwendung der Prüfpräparate schließt sich eine Behandlungsphase mit alleiniger Applikation der Emulsionen ohne weitere Waschungen an. In diesem Modell wurde ein hydrophiles Vehikel mit und ohne Dexpanthenol getestet. Dabei konnte ein hydratisierender und antiinflammatorischer Effekt von Dexpanthenol nachgewiesen werden.

Kinetic study of lipid distribution after washing with lipases: Microscopy analysis

AbstractEffect of lipase on removal of lard from a cotton fabric by detergency was studied in detail by radioisotope and X‐ray microanalysis over 60‐min washing. Osmium tetroxide was used to label the soil for microscopy. Backscattred electron images were used to study residual soil present on and within the cotton fibers and in the interfiber spaces of the yarn bundle. Lard‐soiled samples had large deposits on the fabric surfaces; oil was present in the crenulation, secondary walls, and the lumen of the fibers. Relative concentrations of oil were determined for selected morphological locations within the fiber structure and at the fiber surface using X‐ray microanalysis. Specimens soiled with lard and tagged with osmium tetroxide immediately from the wash bath at 0, 5, 10, 20, and 60 min provide snapshots of the washing process. Within the yarn structure, domains with high oil concentration developed during washing. These domains were not adsorbed onto the fiber surfaces, but were contained within the twisted yarn structure. The inclusion of lipase “accelerates” the washing process with the dislodgment of the soil from the fiber surface being more complete for the yarn specimens washed with lipase for 20 and 60 min. The three main stages of the soil‐removal process—transport of water and detergent to the soil on the substrate, separation of soil and substrate, and transport of dislodged soil into the wash bath—were observed. Lipase functions in detergency by hydrolysis of triglycerides forming mono‐ and diglycerides and fatty acids that are more soluble in water, can undergo soap formation, enhance mesomorphic phase formation, and increase emulsification. The quantification of the mass of the residual oil is well‐defined using the radioisotope methodology. The effectiveness of lipase on removal of lard stains is observed at all points in the washing cycle (5, 10, 20, 60 min). Our data fit a classical kinetic description of the soil‐removal process described by an exponential decay, with the presence of lipase greatly increasing the rate of oil removal during the early period of washing (0 to 20 min). With the use of lipase in the washing bath, the amount of soil removal after 10 min is about the same as observed after washing 60 min with detergent only. Washing removes soil located on the fibers and yarns on the fabric surfaces early in the process and more completely. Soil that is located within the yarn structure is removed by washing but this requires more time, and lard removal from these interfiber capillaries is greatly enhanced by use of lipase. Washing does remove soil from within the cotton fiber, i.e. secondary wall and lumen, and this is also more complete with the use of lipase. After washing with a detergent for 60 min, the highest concentration of residual soil is in the lumen, although the concentration has been reduced by a factor of two. It appears that the lumen is the last region of the cotton fabric structure to be cleaned by laundering. The use of lipase in the wash bath increased the rate of soil removal for all locations with the cotton fabric and fiber structures.