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At least 19 recordsLinked to original sources
PubMed2025

"All doctors should be trained in that": The coproduction and mixed-methods evaluation of an educational toolkit to enable safe, high-quality genetic health care for people with intellectual disability.

PURPOSE: People with intellectual disability inequitably access high-quality genetic health care. However, they are keen to understand genetic health care and recommend that clinicians need education on delivering more inclusive care and that multimodal genetic health literacy resources should be coproduced. METHODS: Our inclusive research team applied best-practice coproduction principles to deliver a suite of resources, the GeneEQUAL Toolkit. Mixed-methods evaluation, including surveys and focus group/interviews, assessed (1) clinicians' perceived capabilities, motivation, and opportunities for providing inclusive health care for people with intellectual disability before and after exploring the Toolkit; (2) the perceptions and opinions of people with intellectual disability about the Toolkit; (3) the reach of the Toolkit components; and (4) the reflections of people with intellectual disability and clinicians on the coproduction process. RESULTS: The Toolkit met the expectations and preferences of people with intellectual disability and clinicians, and had a global reach. Coproduction was feasible and judged as critical for the high value of the Toolkit, in motivating clinicians to change their clinical practice and empowering people with intellectual disability. CONCLUSION: Coproduction can be successfully applied to improve the engagement of people with intellectual disability, potentially reducing health inequity and improving the safety and quality of genetic health care.

Humans↗
PubMed1991

Anticipatory velar lowering: a coproduction account.

Feature spreading and coproduction models make fundamentally different assumptions about the nature and organization of speech motor control, and yet each model is supported by some, but not all, of the existing empirical data. This has led some researchers to conclude that speakers probably use alternative strategies at different times. This study suggests that the identification of coarticulatory influences requires the concurrent identification of intrinsic articulatory characteristics of the segment. Moreover, the evidence for feature spreading or variable coarticulation strategies derives from the misidentification of such intrinsic characteristics as context effects. This velar coarticulation study used a controlled comparison between CVnN and CVnC minimal pairs, where C is an oral consonant, Vn is any number of vowels, and N is a nasal consonant. Vocalic string duration was manipulated by varying the number of segments and speech rate, allowing us to alter the time between the onsets of vocalic and subsequent consonantal gestures. Velar lowering occurred in CVn sequences, whether or not a nasal consonant followed, and similar vocalic gestures were observed across minimally contrastive environments with and without the nasal consonant. Moreover, velar lowering for the nasal consonant began in close temporal proximity to the nasal murmur. These results strongly support the coproduction model and provide insight into previously conflicting reports.

Humans↗
PubMed1990

Coproduction of surfactin and iturin A, lipopeptides with surfactant and antifungal properties, by Bacillus subtilis.

Of the 13 strains of Bacillus subtilis tested for the coproduction of the lipopeptide surfactin and the antifungal lipopeptides of the iturin family, only 1 produced both lipopeptides with a high yield. The cultures were made in a synthetic medium. Several L-amino acids and various carbon sources were good substrates for the lipopeptide production. The maximum yield of surfactin was about 110 mg/liter and that of iturin A about 39 mg/liter/absorbance unit for the best strain, B. subtilis S 499.

Amino Acid Sequence↗
PubMed1976

Coproduction of Regan isoenzyme and carcinoembryonic antigen in HCT-8 cells.

The presence of the Regan isoenzyme of alkaline phosphatase was demonstrated in the carcinoembryonic antigen-producing human cell line HCT-8 Biochemical, histochemical, immunologic, and electrophoretic methods were used as criteria for the identification and characterization of this isoenzyme.

Carcinoembryonic Antigen↗
PubMed1990

Coarticulatory organization for lip rounding in Turkish and English.

A number of studies, involving English, Swedish, French, and Spanish, have shown that, for sequences of rounded vowels separated by nonlabial consonants, both EMG activity and lip protrusion diminish during the intervocalic consonant interval, producing a "trough" pattern. A two-part study was conducted to (a) compare patterns of protrusion movement (upper and lower lip) and EMG activity (orbicularis oris) for speakers of English and Turkish, a language where phonological rules constrain vowels within a word to agree in rounding and (b) determine which of two current models of coarticulation, the "look-ahead" and "coproduction" models, best explained the data. Results showed Turkish speakers producing "plateau" patterns of movement rather than troughs, and unimodal rather than bimodal patterns of EMG activity. In the second part of the study, one prediction of the coproduction model, that articulatory gestures have stable profiles across contexts, was tested by adding and subtracting movement data signals to synthesize naturally occurring patterns. Results suggest English and Turkish may have different modes of coarticulatory organization.

Adult↗
PubMed1992

Phenotypic plasticity of avian embryonic sympathetic neurons grown in a chemically defined medium: direct evidence for noradrenergic and cholinergic properties in the same neurons.

Avian embryonic sympathetic ganglia possess both catecholaminergic and cholinergic features and can synthesize noradrenaline (NAd) and acetylcholine (ACh) simultaneously. In the present study we sought to determine (1) whether or not this coproduction of NAd and ACh corresponds to the existence of two non-overlapping populations, and (2) to what extent the levels of synthesis are influenced by non-neuronal ganglion cells. We have focused on the correlation between the immunocytochemically demonstrable presence of the noradrenergic and cholinergic enzymes tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT), respectively, and the synthesis of the corresponding neurotransmitters in embryonic quail sympathetic neuronal and non-neuronal cells purified by fluorescence-activated cell sorting. We show that (1) freshly sorted neurons synthesize both NAd and ACh, whereas non-neuronal cells produce neither; (2) the overwhelming majority of the sympathetic neurons display TH immunoreactivity; (3) about half of the TH-positive neurons are recognized by an anti-ChAT antibody in an artificial medium that selectively enhances synthesis and/or accumulation of ACh; (4) the non-neuronal cells are important for survival of the neurons and potentiate their synthesis of ACh in this medium, and (5) finally, we present evidence that expression of TH in noradrenergic neurons and in small intensely fluorescent cells of sympathetic ganglia is differentially regulated.

Animals↗
PubMed1987

The inhibition and disposition of intestinal alkaline phosphatase.

A primary mechanism of amino acid inhibition of intestinal alkaline phosphatase is postulated to be the formation of a dissociable enzyme-amino acid complex at an allosteric zinc site. The degree of inhibition was highly correlated with the Zn2+ stability constant of each amino acid and the inhibition was reversible by the addition of exogenous Zn2+ or by dialysis. This allosteric amino acid inhibition proved to be a useful probe of the membrane arrangement of the enzyme in the intact tissue. The catalytic site appears to face the lumen based on the poor permeability of the substrate, the accumulation of the coproducts in the luminal bath, and the response of the enzyme to luminal pH. Amino acid inhibition of alkaline phosphatase in the intact tissue was only effective in the presence of sodium; whereas sodium was not required in butanol extracted preparations which lacked the sidedness of the intact tissue. Since amino acid uptake from the intestine is sodium dependent, the allosteric inhibitory site is probably intracellular. The results suggest that the intestinal alkaline phosphatase spans the apical membrane with the catalytic site accessible from the lumen and the allosteric inhibitory site from the cytoplasm.

Alkaline Phosphatase↗
PubMed1992

Determination of glutamic acid decarboxylase activity and inhibition by an H2O2-sensing glutamic acid oxidase biosensor.

The catalytic activity of the enzyme L-glutamic acid decarboxylase (GAD) is determined by an amperometric method based on a recently developed glutamate-selective biosensor. The biosensor is composed of an amperometric H2O2 electrode and a biocatalytic membrane containing the enzyme glutamic acid oxidase (GAO). The biosensor allows the direct and continuous measurement of GA levels by monitoring the H2O2 produced at the electrode interface as a coproduct of the GAO-catalyzed GA oxidation to alpha-ketoglutaric acid. Since GA is transformed to gamma-aminobutyric acid and CO2 under the catalytic activity of GAD, the rate of GA consumption in solution, monitored by the GAO biosensor, represents a reliable measure of GAD catalytic activity. Additional experiments performed in the presence of different concentrations of the GAD inhibitor valproic acid have shown the suitability of the proposed approach for the study of GAD inhibitors also. Discussion of the main experimental characteristics of this new analytical method is given in terms of sensitivity, reproducibility, and reliability of the experimental results and ease, time, and cost of operation.

Amino Acid Oxidoreductases↗
PubMed1991

Purification and characterization of the monoterpene cyclase gamma-terpinene synthase from Thymus vulgaris.

The monoterpene cyclase, gamma-terpinene synthase, from Thymus vulgaris (thyme) leaves was purified to apparent homogeneity by isoelectric focusing and dye-ligand, anion-exchange, hydrophobic interaction, and gel permeation chromatography. The enzyme has a native molecular weight of 96,000 as determined by gel permeation chromatography, and exhibited a specific activity of 538 nmol/h.mg protein (turnover number of approximately 0.01/s). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the enzyme to be composed of two apparently identical subunits of Mr approximately 55,000. The protein was very hydrophobic, and possessed a pI value of 4.85 as determined by isoelectric focusing. Maximum activity was observed at pH 6.8 in the presence of 20 mM Mg2+; 5 mM Mn2+ could support catalysis, albeit at a much lower rate. The Km value for the substrate, geranyl pyrophosphate, was 2.6 microM. Cyclase activity was inhibited by cysteine- and histidine-directed reagents. Purified gamma-terpinene synthase also possessed the ability to cyclize geranyl pyrophosphate to small amounts of alpha-thujene and to lesser quantities of myrcene, alpha-terpinene, limonene, linalool, terpinen-4-ol, and alpha-terpineol, all of which appear to be coproducts of the reaction sequence leading to gamma-terpinene. In general properties, the gamma-terpinene synthase from thyme leaves resembles other monoterpene cyclases as well as sesquiterpene and diterpene cyclases.

Alkyl and Aryl Transferases↗
PubMed1991

Citrulline in the rat brain: immunohistochemistry and coexistence with NADPH-diaphorase.

The presence in the brain of the urea cycle intermediate citrulline in the absence of a complete urea cycle has never been adequately explained. In an attempt to clarify this problem, we developed antibodies to citrulline and determined the distribution of citrulline-immunoreactivity in fixed sections of rat brain using immunoperoxidase and indirect immunofluorescence techniques. Citrulline-positive neurons were found to have a restricted distribution within the brain. A few cells were present in the cortex and corpus callosum. A large population of strongly stained cells was diffusely scattered throughout the striatum, nucleus accumbens and olfactory tubercle. Less strongly stained cells were detected in the supraoptic and paraventricular nuclei of the hypothalamus, the dorsal raphe, and the laterodorsal and pedunculopontine tegmental nuclei of the pons. The citrulline-immunoreactive cells were similar to those previously shown to contain NADPH-diaphorase activity, and double staining experiments indicated that citrulline-immunoreactivity was present in a subpopulation of NADPH-diaphorase-positive neurons. We have recently identified NADPH-diaphorase as a nitric oxide synthase. Thus the presence of citrulline in these cells suggests that it is formed within the brain as a coproduct during nitric oxide formation from arginine.

Amino Acid Oxidoreductases↗
PubMed1988

Mechanism of monofunctional and bifunctional alkylation of DNA by mitomycin C.

The relative amounts of monofunctional and bifunctional alkylation products of DNA with mitomycin C (MC) depend on whether one or both masked alkylating functions of MC are activated reductively; adduct 8 is the result of one function and adducts 7 and 9, formed as a pair, are the result of both functions being activated [Tomasz, M., Lipman, R., Chowdary, C., Pawlak, J., Verdine, G. L., & Nakanishi, K. (1987) Science (Washington, D.C.) 235, 1204-1208]. To determine the mechanism governing this differential reactivity of MC with DNA, MC-Micrococcus luteus DNA complexes formed under varying conditions in vitro were digested to nucleosides and adducts. Adduct distribution, analyzed by high-performance liquid chromatography, served as the measure of monofunctional and bifunctional activation. H2/PtO2 and xanthine oxidase/reduced nicotinamide adenine dinucleotide (NADH) activated MC mostly monofunctionally, and Na2S2O4 activated the drug bifunctionally under comparable conditions. Excess MC selectively suppressed, but excess PtO2 selectively promoted, bifunctional activation by H2/PtO2; excess xanthine oxidase and/or NADH also had promoting effects. O2 tested in the Na2S2O4 system was inhibitory. 10-Decarbamoyl-MC acted strictly monofunctionally under all conditions. Monoadducts bound to DNA were converted to bis adducts upon rereduction. A mechanism with the following features was derived: (i) Activation of MC at C-1 and C-10 is sequential (C-1 first). (ii) A one-time reduction is sufficient for both. (iii) Activation of the second function may be selectively inhibited by kinetic factors or O2. (iv) 7 and 9 are coproducts of bifunctional activation; their ratio depends on the DNA base sequence. (v) Activation of the second function involves an iminium intermediate. Direct applications to the action of MC in vivo are discussed.

Alkylation↗
PubMed1987

The role of coarticulatory effects in the perception of fricatives by children and adults.

Adult listeners are sensitive to the acoustic variations that result from a speaker's coarticulation (or coproduction) of phonetic segments. The present study charted the development of such sensitivity in young children by examining their responses to coarticulatory effects in fricative-vowel syllables. Children, at each of the ages 3, 4, 5, and 7 years, and adults identified tokens from a synthetic /sh/-/s/ continuum followed by one of four natural vocalic portions: /i/ and /u/, produced with transitions appropriate for either /sh/ or /s/. Children demonstrated larger shifts in fricative phoneme boundaries as a function of vocalic transition than did adults, but relatively smaller shifts as a function of vowel quality. Responses were less consistent for children than for adults, and differences between children and adults decreased as children increased in age. Overall, these results indicate that perceptual sensitivity to certain coarticulatory effects is present at as young as 3 years of age. Moreover, the decrease in the sensitivity to vocalic transitions with age suggests that, contrary to a commonly held view, the perceptual organization of speech may become more rather than less segmental as the child develops.

Adult↗
PubMed1991

Purification of a soluble isoform of guanylyl cyclase-activating-factor synthase.

The soluble form of guanylyl cyclase-activating-factor (GAF) synthase from rat cerebellum was purified to homogeneity by sequential affinity chromatographic steps on adenosine 2',5'-bisphosphate (2',5'-ADP)-Sepharose and calmodulin-agarose. Enzyme activity during purification was bioassayed by the L-arginine-, NADPH-, and Ca2+/calmodulin-dependent formation of a plasma membrane-permeable nitric oxide-like factor that stimulated soluble guanylyl cyclase in RFL-6 cells. With calmodulin and NADPH as cofactors, purified soluble GAF synthase induced an increase of 1.05 mumol of cGMP per 10(6) RFL-6 cells per 3 min per mg of protein. The coproduct of this signal-transduction pathway appeared to be L-citrulline. GAF synthase catalyzed the conversion of 107 nmol of L-arginine into L-citrulline per min per mg of protein. Based on these assays, this represents a purification of GAF synthase of approximately 10,076- and 8925-fold with recoveries of 16% and 19%, respectively. Rechromatography of the purified enzyme on Mono P (isoelectric point = 6.1 +/- 0.3), Mono Q, and Superose 12 or 6 resulted in no further purification or increase in specific activity. A Stokes radius of 7.9 +/- 0.3 nm and a sedimentation coefficient s20,w of 7.8 +/- 0.2 S were used to calculate a molecular mass of about 279 +/- 25 kDa for the native enzyme. SDS/PAGE revealed a single protein band with a molecular mass of about 155 +/- 3 kDa. These data suggest that soluble GAF synthase purified from rat cerebellum is a homodimer of 155-kDa subunits and that enzyme activity is dependent upon the presence of calmodulin.

Amino Acid Oxidoreductases↗
PubMed2026

Yeast Strain Development and Process Intensification in High-Gravity Fermentation.

High- and very-high-gravity (HG/VHG) fermentation increases substrate loading and product titers, thereby improving fermenter utilisation and potentially reducing water use and downstream processing requirements. Initially developed for brewing and fuel ethanol production, these approaches are now applied more broadly in food, beverage, and bioproduct manufacturing. This MiniReview summarises operational definitions and industrial drivers of HG/VHG fermentation and examines the associated constraints in rheology, mass and heat transfer, osmotic and ethanol stress, nutrient availability, and oxidative damage. Yeast improvement strategies are reviewed, including adaptive laboratory evolution, mutagenesis, genome shuffling, multiplex genome editing, non-conventional yeasts, and multi-omics-guided selection. Process developments such as no-cook simultaneous liquefaction, saccharification and fermentation (SLSF), enzyme formulation, nutrient management, and in situ product recovery are considered together with applications in alcoholic beverages, organic acids, microbial lipids, and other value-added products. The review also discusses coproduct valorisation and the need to integrate strain development with process design. Current evidence supports HG/VHG fermentation as a useful process-intensification platform, although performance and sustainability depend strongly on feedstock, operating conditions, product requirements, and the basis used to report fermentation outcomes.

circular bioeconomy↗
PubMed1990

Neonatal necrotizing enterocolitis associated with delta toxin-producing methicillin-resistant Staphylococcus aureus.

Delta toxin-producing coagulase-negative staphylococci previously have been associated with necrotizing enterocolitis in neonates. We identified three preterm infants (body weight, 845 +/- 59 g) infected with methicillin-resistant Staphylococcus aureus (MRSA) who had a similar clinical syndrome, characterized by pustular dermatitis, bacteremia and necrotizing enterocolitis accompanied by gastric residua, abdominal distention, hematochezia and pneumatosis intestinalis. MRSA was recovered from all three infants at infected skin sites, blood or venous catheters and from two of three infants in stool specimens. Two infants also had Staphylococcus epidermidis isolated from stool. All MRSA isolates had identical microbiologic profiles: four plasmids with identical molecular weights; coproduction of enterotoxins A and B; and the same antibiotic susceptibilities. Of one skin isolate, two blood isolates and two stool isolates of MRSA that were tested, all had characteristic delta toxin hemolytic activity. All culture supernatants of these isolates evaluated for delta toxin were positive by Western blot analysis. The two strains of S. epidermidis isolated from stool were negative for delta-like toxin by a standardized enzyme-linked immunoassay. The clustering of these cases, the similarity of the clinical syndrome, and the prior association of necrotizing enterocolitis with delta-like toxins produced by S. epidermidis, suggest that delta toxin-producing MRSA (or other S. aureus isolates) also may be etiologic agents in some cases of necrotizing enterocolitis in newborns.

Bacterial Toxins↗
PubMed1991

Hydrated electron formation on laser excitation of P-pyridoxyl amino acids and proteins.

Evidence from a characteristic transient spectrum, its lifetime and quenching by N2O demonstrates that XeCl excimer laser excitation of the reduced form of pyridoxal 5'-phosphate bound to lysine residues will produce the hydrated electron. The phenoxyl-type radical coproduct was also observed. The results identify a useful label of protein structures for site-specific electron generation which can occur outside the region of light absorption by aromatic residues of protein. The results also represent the first observation of hydrated electron formation from flash excitation of a vitamin B6 cofactor.

Electrons↗
PubMed1988

Leucine auxotrophy specifically alters the pattern of trichothecene production in a T-2 toxin-producing strain of Fusarium sporotrichioides.

The biosynthetic pathway for trichothecenes in the filamentous fungus Fusarium sporotrichioides NRRL 3299 has been further characterized. Experiments using the techniques of mutational analysis and the incorporation of radiolabeled precursors indicated that leucine is a direct precursor to the isovalerate moiety present in the trichothecene, T-2 toxin. Analysis of trichothecene production in a UV-induced leucine auxotroph also revealed the existence of a branched biosynthetic pathway which results in the coproduction of T-2 toxin and the T-2 toxin analogs neosolaniol, 8-isobutyryl-neosolaniol, and 8-propionyl-neosolaniol. Leucine limitation imposed by the leucine auxotroph simultaneously led to underproduction of T-2 toxin and overproduction of these T-2 toxin analogs, which are produced in small amounts by the wild-type parent. Furthermore, it was shown that the ratio of T-2 toxin to T-2 toxin analogs produced by the leucine auxotroph can be modulated by the concentration of leucine in the medium. These results suggest that the four trichothecenes mentioned above are derived from a common intermediate and that there is competition for this intermediate among the branched pathways leading to these four cometabolites.

Chemical Phenomena↗
PubMed1992

Ethanol production from cellobiose, amorphous cellulose, and crystalline cellulose by recombinant Klebsiella oxytoca containing chromosomally integrated Zymomonas mobilis genes for ethanol production and plasmids expressing thermostable cellulase genes from Clostridium thermocellum.

The Zymomonas mobilis genes for ethanol production have been integrated into the chromosome of Klebsiella oxytoca M5A1. The best of these constructs, strain P2, produced ethanol efficiently from cellobiose in addition to monomeric sugars. Utilization of cellobiose and cellotriose by this strain eliminated the requirement for external beta-glucosidase and reduced the amount of commercial cellulase needed to ferment Solka Floc SW40 (primarily crystalline cellulose). The addition of plasmids encoding endoglucanases from Clostridium thermocellum resulted in the intracellular accumulation of thermostable enzymes as coproducts with ethanol during fermentation. The best of these, strain P2(pCT603T) containing celD, was used to hydrolyze amorphous cellulose to cellobiose and produce ethanol in a two-stage process. Strain P2(pCT603T) was also tested in combination with commercial cellulases. Pretreatment of Solka Floc SW40 at 60 degrees C with endoglucanase D substantially reduced the amount of commercial cellulase required to ferment Solka Floc. The stimulatory effect of the endoglucanase D pretreatment may result from the hydrolysis of amorphous regions, exposing additional sites for attack by fungal cellulases. Since endoglucanase D functions as part of a complex in C. thermocellum, it is possible that this enzyme may complex with fungal enzymes or bind cellulose to produce a more open structure for hydrolysis.

Cellobiose↗