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Biosynthesis of Alkali Insoluble Polysaccharide from UDP-d-Glucose with Particulate Enzyme Preparations from Phaseolus aureus.

A particulate enzyme system from Phaseolus aureus seedlings catalyzes the synthesis of alkali insoluble polysaccharide material from UDP-d-glucose. 80 to 90% of the d-glucose units are joined by beta-1,4 linkages, the remainder being combined by beta-1,3 linkages. It is not known whether the material is a single polysaccharide or a mixture.With a substrate concentration of 4 x 10(-3)m the enzyme system catalyzes the formation of alkali insoluble polysaccharide from UDP-d-glucose at a rate 23 times greater than that from GDP-d-glucose. The modifications introduced also increase the formation of cellulose from GDP-d-glucose to 3 times the rate previously reported.None of the glycosyl nucleotides containing bases other than guanine or uracil (adenine, cytosine, thymine) served as substrate for the production of alkali insoluble polysaccharide with this enzyme system. dUDP-d-glucose could also not serve as substrate.

Journal Article↗

Alkali Cation/Sucrose Co-transport in the Root Sink of Sugar Beet.

The mechanism of sucrose transport into the vacuole of root parenchyma cells of sugar beet was investigated using discs of intact tissue. Active sucrose uptake was evident only at the tonoplast. Sucrose caused a transient 8.3 millivolts depolarization of the membrane potential, suggesting an ion co-transport mechanism. Sucrose also stimulated net proton efflux. Active (net) uptake of sucrose was strongly affected by factors that influence the alkali cation and proton gradients across biological membranes. Alkali cations (Na(+) and K(+)) at 95 millimolar activity stimulated active uptake of sucrose 2.1- to 4-fold, whereas membrane-permeating anions inhibited active sucrose uptake. The pH optima for uptake was between 6.5 and 7.0, pH values slightly higher than those of the vacuole. The ionophores valinomycin, gramicidin D, and carbonyl cyanide m-chlorophenylhydrazone at 10 micromolar concentrations strongly inhibited active sucrose uptake. These data are consistent with the hypothesis that an alkali cation influx/proton efflux reaction is coupled to the active uptake of sucrose into the vacuole of parenchyma cells in the root sink of sugar beets.

Journal Article↗

Laser scanning confocal microscopy for in situ monitoring of alkali-silica reaction.

Alkali-silica reaction (ASR) occurs in concrete between reactive siliceous components in the aggregate and the strongly alkaline pore solution, resulting in the formation of a potentially expansive gel product. Lithium additives have been shown to reduce expansion associated with ASR, but the mechanism(s) by which lithium reduces expansion have not been understood. Therefore, development of an in situ method to observe reactions associated with ASR is highly desirable, as it will allow for non-destructive observation of the reaction product formation and damage evolution over time, as the reaction progresses. A technique to image into mortar through glass aggregate by laser scanning confocal microscopy (LSCM), producing three-dimensional representations of the sample was developed. This LSCM technique was utilized to monitor the progress of alkali-silica reaction in mortar samples prepared with alkali-reactive glass aggregate both in the presence and in the absence of lithium additives: LiNO3, LiCl or LiOH. The method proved to be effective in qualitatively monitoring crack formation and growth and product formation, within cracks and at the paste/aggregate interface. In particular, dendritic products were observed at the paste/aggregate interface only in those samples containing lithium, suggesting that these products may play a role in ASR mitigation.

Journal Article↗

Studies on the actomyosin ATPase and the role of the alkali light chains.

Myosin isoenzymes, highly enriched in either alkali 1 or alkali 2 light chains have been prepared by light chain exchange in 4.7 M ammonium chloride, under conditions where there is minimal loss of ATPase activity. While the actin-activated ATPase measurements were complicated by a biphasic dependence on actin concentration, the two myosin isoenzymes behaved in a similar manner; at a variety of ionic strength conditions their maximum rates of ATP hydrolysis were nearly identical. Furthermore, under conditions where their Km values could be reliably determined, their apparent affinities for actin in the presence of ATP did not differ greatly. These results suggest that the presence of a particular alkali light chain does not influence the maximum rate of ATP turnover by actomyosin under ionic strength conditions approximating physiological.

Actins↗

Alkali citrate prophylaxis in idiopathic recurrent calcium oxalate urolithiasis--a prospective randomized study.

OBJECTIVE: To assess the efficacy of alkali citrate therapy in patients suffering from recurrent idiopathic calcium oxalate stone formation. PATIENTS AND METHODS: Fifty patients suffering from active stone formation who had had at least one stone annually over the previous 3 years were recruited and randomly divided into two groups. They were given either general prophylactic instructions, i.e. abundant liquid intake and dietary restrictions (group I, n = 25)--or were additionally treated with alkali citrate (group II, n = 25). RESULTS: In group I, the rate of stone formation was reduced from 1.8 to 0.7 stones per patient per year. Similar results were obtained for group II, the corresponding figures being 2.1 and 0.9. There was a statistically significant higher urinary citrate excretion in group II than in group I. No difference was seen between the two groups regarding recurrent stone formation (Student's t-test). Stone formation decreased in both groups (group I 27%, group II 31%). Subjectively, 56% of group II patients, all of whom had previously experienced severe colic, reported spontaneous stone elimination to be painless, whereas in group I all but one patient suffered pain. CONCLUSION: An objective benefit of alkali citrate could not be established.

Adult↗

Transplantation of reconstructed corneal layer composed of corneal epithelium and fibroblasts on a lyophilized amniotic membrane to severely alkali-burned cornea.

The purpose of this article was to evaluate the graft efficacy of reconstructed corneal layer, composed of autologous corneal epithelium and fibroblasts on a lyophilized amniotic membrane (LAM), in a severely alkali-burned corneal model. After biopsy specimens were obtained from the left eyes of 24 rabbits, the corneal epithelial cells and fibroblasts were expanded in vitro and the corneal layer was reconstructed on LAM. Thirty-six eyes of rabbits underwent alkali burn (1 N NaOH, 30 s) to create a limbal deficiency and a deeply damaged corneal stroma. Four weeks later, group 1 underwent a graft of the reconstructed corneal layer composed of autologous corneal epithelium and fibroblasts on LAM. Group 2 was transplanted with a graft of the reconstructed autologous corneal epithelium, and group 3 served as a control without surgery. Wound healing and stabilization of the ocular surfaces occurred much faster in group 1 than in groups 2 and 3. The eyes in group 3 revealed typical limbal deficiencies with conjuctivalization and persistent corneal epithelial defects. However, the corneas in group 1 developed only mild peripheral neovascularization. Immunohistochemical staining in group 1 demonstrated that p63, cytokeratin 3, E-cadherin, transforming growth factor (TGF)-beta1, and collagen IV were expressed strongly in the corneal epithelium and basement membrane. On the basis of these results, transplantation of the reconstructed corneal layer, composed of autologous corneal epithelium and fibroblasts on LAM, partially accelerated the recovery of the alkali-injured rabbit ocular surface, and might be useful therapeutically for the treatment of patients with severely damaged cornea.

Amnion↗

Thermal effects of neutralization therapy and water dilution for acute alkali exposure in canines.

OBJECTIVE: To evaluate the change in temperature of the gastric mucosa and lumen contents when a weak acid or water is used to manage acute alkali exposure. METHODS: A prospective in-vivo canine model was used in a university-based animal laboratory setting. Eighteen adult canines weighing 20-25 kg were placed under a surgical plane of anesthesia and a laparotomy was performed. A gastrotomy was then made later ligation of the distal esophagus and proximal duodenum. Separate mucosa and lumen temperature probes were placed. Then 25 mL of room-temperature (24-26 degrees C) 50% sodium hydroxide (NaOH) was instilled in the gastric lumen. After 5 minutes, each canine was given treatment. Group 1 (n = 10) was treated with 75 mL of room-temperature orange juice. Group 2 (n = 8) was treated with 75 mL of room-temperature water. Continuous mucosa and lumen temperatures were observed and recorded at baseline and at specified intervals for 35 minutes after the alkali insult. Repeated-measures analysis of variance was used to evaluate the overall temperature profile. Signed-rank tests were used to compare the changes in temperature immediately following neutralization treatment. RESULTS: Significant temperature decreases of 1.1 degrees C and 2.1 degrees C were observed for both mucosa (p = 0.002) and lumen (p < 0.001) temperature, respectively, following neutralization therapy with room-temperature orange juice. In the group treated with room-temperature water, significant temperature decreases of 2.1 degrees C for mucosa (p = 0.01) and 2.4 degrees C for lumen (p = 0.01) were observed. Posttreatment temperatures did not exceed baseline for the entire observation period. CONCLUSION: Neutralization therapy with room-temperature orange juice or water dilution for acute gastric injuries by liquid alkali does not cause a rise in mucosal or intraluminal temperatures in an in-vivo canine model.

Acids↗

Corneal alkali burn in the rabbit. Waterbalance, healing and transparency.

Healing following a standardized central corneal alkali wound was studied morphologically in New Zealand white rabbits up to one year after the initial wound. Clinical examination, light and transmission electron microscopy was performed. The study was focused on how permanent scar tissue formed. Following the penetrating alkali injury, all cells (epithelium, keratocytes and endothelium) in the wound area disappeared. The fibroblasts/keratocytes repopulated an extensively swollen central corneal stroma. Cells and extracellular matrix filled stromal lacunae in an irregular fashion and upon deswelling the lacunae remained as irregularities in the stroma, reducing the transparency. In the periphery of the wound repopulation occurred in a less swollen stroma and normal cytoarchitecture and transparency resumed. It appears that the degree of swelling decides the degree of scar tissue formation in the corneal stroma following alkali wound healing.

Animals↗

Unloaded shortening velocity and myosin heavy chain and alkali light chain isoform composition in rat skeletal muscle fibres.

1. This study aims to assess the role of myosin heavy chain (MHC) and alkali myosin light chain (MLC) isoforms in determining maximum velocity of shortening in fast skeletal muscle fibres. 2. The maximum velocity of shortening as determined by the slack test (Vo) was tested for its relationship with MHC composition and with alkali MLC isoform ratio of fast fibres of known MHC composition. 3. MHC isoform composition was determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and monoclonal antibodies against MHCs, and combining the results obtained using the two methods. Three groups of fast fibres containing only one MHC isoform were identified: IIA, IIX and IIB fibres containing respectively IIA MHC, IIX MHC and IIB MHC. Fibres containing more than one MHC isoform were discarded. 4. The mean Vo value of IIA fibres was 2.33 +/- 0.29 muscle lengths per second (L s-1; mean +/- S.D.), this was significantly lower than that for IIX fibres (3.07 +/- 0.70 L s-1) which in turn had a mean Vo value significantly lower than that for IIB fibres (3.69 +/- 1.01 L s-1). 5. The relative proportion of alkali MLC isoforms (MLC3f, MLC1f) was determined by means of electrophoretic separation and densitometric quantification and was expressed as MLC3f/MLC2f with reference to the dithio-nitrobenzoic acid (DTNB) light chain (MLC2f). The mean value of the MLC3f/MLC2f ratio was significantly lower in IIA than in IIX and IIB fibres. 6. Vo was found to be proportional to the relative content of MLC3f in IIA, IIX and IIB fibres.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Germination of heat- and alkali-altered spores of Clostridium perfringens type A by lysozyme and an initiation protein.

The normal system functioning in the utilization of metabolizable germinants by both heat-sensitive and heat-resistant spores of Clostridium perfringens was inactivated by heat or by treatment of the spores with alkali to remove a soluble coat protein layer. Altered spores were incapable of germination (less than 1%) and outgrowth (less than 0.0005%) in complex media without the addition of either lysozyme or an initiation protein produced by C. perfringens. The addition of either of these agents permitted, in the case of alkali-treated spores, both 90 to 95% germination and outgrowth, as measured by colony formation. In the case of heat-damaged spores, only 50% germination and 2% outgrowth resulted from addition of the initiation protein, whereas lysozyme permitted 85% germination and 8% outgrowth. Alteration of the spores by heat or alkali apparently inactivated the normal lytic system responsible for cortical degradation during germination. Kinetics of production of the initiation protein and conditions affecting both its activity and that of lysozyme on altered spores are described.

Bacterial Proteins↗

Lack of efficacy of alkali treatment for isolation of Yersinia enterocolitica from feces.

The efficacy of KOH treatment to enhance isolation of Yersinia enterocolitica from stools containing fecal flora which is not adequately suppressed by selective media was tested by direct plating and enrichment techniques. Although a distinct difference in the level of alkali tolerance was observed between Y. enterocolitica and the fecal organisms included in the study, the selective suppression of the fecal flora from stools could not be consistently achieved on this basis. By direct plating, the isolation rates ranged from 4 to 32% for untreated control samples and 11 to 40% for KOH-treated samples, varying with the medium used for culture. The maximum overall recovery rate was 34% by overnight enrichment of fecal samples in Selenite F, tetrathionate, Trypticase soy, and gram-negative broths or by 2-day enrichment in modified Rappaport broth. Alkali treatment of fecal samples did not significantly enhance the isolation rates by these methods. One hundred percent recovery was achieved by prolonged cold enrichment, and alkali treatment made no difference in the recovery rate by this method. Cefsulodinirgasan-novobiocin agar generally yielded higher isolation rates of Y. enterocolitica than did cellobiose-arginine-lysine agar and MacConkey agar, both by direct plating of fecal samples and through subcultures of enrichments, regardless of KOH treatment.

Chemical Phenomena↗

Acid production in diabetic acidosis; a more rational approach to alkali replacement.

The production of organic acids in severe diabetic acidosis was studied to determine the contribution of various acids and to reassess alkali requirements. In 11 patients the mean total concentration of determined organic acids was 16 mEq/l., while the mean estimated base deficit was 24 mEq/l. Acetoacetic and beta-hydroxybutyric acids accounted for 75% of measured organic acid. In 10 patients the mean amount of sodium bicarbonate administered for correction of the acidosis was 185 mEq, while the mean requirement was 394 mEq.These findings imply that the methods commonly used to determine the base deficit and the alkali requirements in patients with diabetic acidosis may be invalid. The prompt administration of alkali should be limited, and we suggest that the blood pH should be restored only to 7.25.

Acetoacetates↗

Raised parathyroid hormone levels in the milk alkali syndrome: an appropriate response?

A case of the 'milk alkali syndrome' associated with grossly elevated levels of amino terminal parathyroid hormone is described. The hypercalcaemia (calcium 4.09 mmol/l) and hyperparathyroidism settled on conservative measures. Factors in the milk alkali syndrome which might stimulate the release of parathyroid hormone include parathyroid gland hyperplasia secondary to suppression of ionized calcium, alteration in sensitivity of calcium receptors on the cells of the parathyroid glands, the stimulation of an intermittent alkaline tide in the blood and the high intake of phosphate and bicarbonate. We suggest that high levels of parathyroid hormone in the milk alkali syndrome may be appropriate rather than paradoxical.

Adult↗

Plasminogen kringle 5 inhibits alkali-burn-induced corneal neovascularization.

PURPOSE: Plasminogen kringle 5 (K5) is a potent angiogenic inhibitor. The purpose of the present study was to evaluate the therapeutic effect of K5 on alkali-burn-induced corneal neovascularization (NV) and to investigate its mechanism of action. METHODS: Corneal NV was induced in rabbits by NaOH. The rabbits received eye drops containing K5 or vehicle alone, four times per day. Corneal NV and inflammation were monitored every other day with a slit lamp microscope, and the length of the vessels in the cornea and the area of NV were measured. Vascular endothelial growth factor (VEGF) was determined by immunohistochemical and Western blot analyses. The TUNEL assay was used to assess the apoptosis of endothelial cells. The effects of K5 on primary bovine aortic endothelial cells (BAECs) were determined by MTT assay, flow cytometry, transmission electron microscopy, and DNA fragmentation assay. RESULTS: Alkali-burn-induced progressive corneal NV and inflammation in the cornea. K5 delayed the onset of corneal NV (P < 0.05) and decreased NV areas (P < 0.05) in a dose-dependent manner. K5 treatment, after the formation of corneal NV, induced regression of newly formatted vessels in the cornea. K5 decreased the inflammatory index in the corneas at different time points after the alkali burn. Corneal VEGF levels were reduced by K5 treatment. K5 inhibits proliferation and induces apoptosis in BAECs. CONCLUSIONS: Topical application of K5 may have therapeutic potential for the chemical burn-induced corneal NV and inflammation. The inhibitory effect of K5 on corneal NV may be by downregulation of VEGF expression.

Administration, Topical↗

Expression of K12 keratin in alkali-burned rabbit corneas.

The healing of alkali-injured corneas is characterized by the persistence of polymorphonuclear leukocytes (PMN) in tissues and recurrent corneal epithelial defects. It has been suggested that the proteolytic enzymes secreted by PMN may account in part for the recurrent epithelial defects in the alkali-burned corneas. Cytoplasmic keratins, which form intracellular intermediate filaments, participate in the formation of hemidesmosomes and play a key role in the focal adhesion of epithelial cells to the basement membranes. The K3/K12 keratin pair is a major constituent of differentiated and stratified corneal epithelium. We have recently cloned the cDNA encoding the rabbit K12 keratin. In the present study we examined the expression of K12 keratin during the healing of alkali-burned rabbit corneas by slot-blot and in situ hybridization. Our results indicate that in normal cornea K12 keratin is equally expressed in all cell layers of stratified corneal epithelium and suprabasal layers of limbal epithelium, but not in bulbar conjunctival and other epithelia, i.e., lens, iris, and retinal pigment epithelium. The basal cells of the detached regenerating epithelium of the injured cornea express a very low level of K12 keratin. These observations are consistent with the notion that defective expression of K3/K12 keratins may play a role in the abnormal attachment of the regenerating epithelium to the basement membrane.

Animals↗

Trauma and alkali burns induce distinct patterns of cytokine gene expression in the rat cornea.

Cytokines such as the interleukins (IL) and tumor necrosis factor alpha (TNF alpha) have traditionally been associated with paracrine regulation of immune reactions. These proteins also have properties suggestive of functional roles in the inflammatory and reparative responses to tissue injury. In this study, mRNA levels for IL-1 alpha, IL-1 beta, IL-6, TNF alpha, interferon gamma, transforming growth factor beta 1, and CD4 were monitored in rat corneas at times from 1 hour through 2 weeks after incisional trauma or alkali burns. Transcripts for IL-1 alpha, TNF alpha, and TGF beta 1 were present in most corneal samples; whereas those for IFN gamma and CD4 were not detected. As early as 1 hour following either of these non-immunologic forms of injury, expression of IL-6 mRNA levels was induced. Only in corneas with alkali burns did IL-6 induction persist from days 1 through 7. The alkali-injured corneas also had markedly increased IL-1 beta mRNA levels from days 1 through 7. These observations indicate that cytokine mRNA is induced in the cornea by trauma without an apparent immunologic stimulus. Our data are consistent with the hypothesis that corneal tissues respond to different types of injury with different patterns of cytokine gene expression.

Animals↗

Disappearance of hemicellulosic monosaccharides and alkali-soluble phenolic compounds of normal and brown midrib sorghum x sundangrass silages fed to Holstein steers.

Four ruminally cannulated Holstein steers fed ad libitum were used in a 4 x 4 Latin square design. Feeds were normal and brown midrib genotypes of Redlan x Piper and Redlan x Greenleaf sorghum-sudangrasses. An in situ trial was run in conjunction with the digestibility trial as part of the data collection for each period. Hemicellulosic monosaccharide and alkali-soluble lignin phenolic compound disappearances were quantitated. The xylose to arabinose ratio was higher in normal genotypes than in brown midribs. Normal genotypes had higher concentrations of alkali-soluble p-coumaric acid and lower vanillin, vanillic acid, and sinapic acid than did brown midrib mutants. Disappearance of p-coumaric acid was higher in steers consuming normal genotypes than in those consuming brown midribs. Neutral detergent fiber, acid detergent fiber, hemicellulose, cellulose, and acid detergent lignin showed no genotypic differences in rate of digestion, but extent of 72 h digestion was higher for brown midrib mutants than for normal genotypes. Extent of digestion of the hemicellulosic monosaccharides, xylose, and uronic acids was higher in situ for brown midrib silages compared with normal genotypes. Differences in alkali-soluble phenolic compound and cell wall monosaccharide profiles between normal and brown midrib genotypes may help explain digestibility differences between the two genotypes.

Animal Feed↗

Effect of topically applied Na-hyaluronan on experimental corneal alkali wound healing.

The effect of topically applied 1% sodium hyaluronate (Na-HA) on the healing of a standardized corneal alkali wound was studied. The healing of the epithelium, stroma, and endothelium was evaluated separately, using quantitative methods. Central corneal alkali wound was produced in one eye of the rabbits by applying a 5.5-mm round filter paper, soaked in 1 N NaOH, for 60 seconds. 1% Na-HA in the treatment group and phosphate buffered saline (PBS) in the control group were given topically 4 times per day for 2 days, 1- and 3-weeks. Epithelial and endothelial healing was assessed morphometrically from standardized photographs and micrographs, respectively. Stromal healing was determined by counting polymorphonuclear leukocytes (PMN) and keratocytes in the central and marginal wound areas. A positive healing influence was observed in the epithelium. In stromal healing, 1% Na-HA treated corneas showed less PMNs and more keratocytes than the control group, suggesting that topically applied 1% Na-HA may suppress the stromal PMN infiltration and enhance the keratocyte repopulation during corneal alkali wound healing. However, no significant difference was found in morphometric evaluation of endothelial healing between the two groups.

Administration, Topical↗