Glucose Assimilation Research Articles

Biocatalyst preparation from Pseudomonas putida SM-6 for conversion of dl-lactate to pyruvate

The production of pyruvate using enzymatic methods is a valuable process for its cheap substrate, high conversion ratio, simple composition of reaction, and convenience of recovery. Cell-free extracts from Pseudomonas putida SM-6 that contain both lactate oxidase (LOD) and catalase were employed as catalysts for biocatalysis. In order to obtain an efficient and cost-effective biocatalyst, the cultivation of P. putida SM-6 both in shake flask and at bioreactor level was investigated. On enzyme production, the suitability of glucose instead of lactate as a carbon source was studied. Maximum LOD production occurred when a mixture of lactate and glucose was used as a carbon source. Lactate was assimilated first and the increase of nitrogen concentration had a positive effect on cell growth and glucose assimilation. The optimum cultivation temperature was 29–31 °C, and the neutral or basic initial medium pH was in favor of both cell growth and LOD production. The interrelation between various process parameters was evaluated for the cultivation of P. putida SM-6 in submerged culture. Using a fed-batch cultivation strategy in a 2-l bioreactor, an increase in cell mass was achieved but without a concomitant increase in LOD production compared with cultivation in shake flask. Bioconversion of lactate to pyruvate was carried out using cell-free extract acquired by cell disruption of P. putida SM-6. Ethylenediaminetetraacetic acid (EDTA) was added into the biocatalytic system to prohibit the degradation of pyruvate by other enzymes in the cell-free extract. As a result, on a preparative scale, cell-free extract which contained 3.04 mg/ml crude enzyme (10.0 U/ml LOD, 877 U/ml catalase) produced pyruvate of 423 mM from dl-lactate of 660 mM in a reaction period of 22 h. The molar conversion yield of pyruvate on the basis of the amount of lactate consumed was 0.80 g/g.

Modulation of the Post‐Prandial Plasma Glucose Profile in Juvenile European Sea Bass Dicentrarchus labrax Fed Diets Varying in Starch Complexity

AbstractThe post‐prandial glycemic response of juvenile European sea bass (initial mean weight 10 g) was determined following acclimation to experimental diets that contained a pea seed meal included at two levels. The ingredient provided a source of starch that replaced purified corn starcWyellow dextrin (2:1) in a reference diet based on fishmeal as the main protein component. The total carbohydrate levels amounted to 25% of the dry matter content and contributed (18%) to the overall gross dietary energy value of 20 MJ/kg. After a 12‐wk feeding trial, in which the sea bass grew to over 40 g (mean weight), the plasma glucose profile was recorded over a 24‐h period (0 h, 1 h, 3 h, 6 h, 9 h, 12 h, and 24 h) following a meal fed to satiation. Results showed that sea bass are able to regulate their glucostatic tolerance to dietary starch, and that the pea seed meal modulated this effect. Peak post‐prandial responses varied between 1 to 9 h and were significantly delayed for the reference diet. Efficacy of glucose assimilation improved for the highest level of pea seed meal inclusion (40% by weight). The relevance of carbohydrate complexity with respect to feed formulation and nutritional requirements of sea bass are discussed.

Digestive enzymes of leaf-cutting ants, Acromyrmex subterraneus (Hymenoptera: Formicidae: Attini): distribution in the gut of adult workers and partial characterization

Enzyme activities associated with the labial glands, midgut and rectum of adult Acromyrmex subterraneus were investigated in order to understand their role in digestion of plant and fungal material. High chitinolytic activity was detected in the labial glands, indicating a possible role in the degradation of fungus ingested by the ants. Chitinolytic activity seen in other compartments of the alimentary canal probably originated in the labial glands. The highest activity detected in the midgut was for α-glucosidase, which was considered to be of insect origin due to its association with midgut epithelium and it is probably involved in glucose assimilation from nutrient sources such as maltose and sucrose present in plant material. A large range of enzyme activities were detected in the rectal lumen contents, and as in the midgut the highest values were for α-glucosidase activity. The absence of activity associated with the epithelium, in the particulate fraction, indicates that the rectal epithelium does not have a secretory function. The detection of enzymes in the rectal lumen contents, which were not detected in the midgut lumen contents, indicates that the rectum acts as a reservoir, accumulating enzymes. The major digestive enzymes were partially characterized using hydrophobic interaction chromatography, gel filtration and SDS-PAGE. The pH of the adult intestinal tract and flow rate of dye through the tract was investigated. A gradual acidification of the intestinal tract was noted commencing with the crop (pH 6−8.2) and terminating with the rectum (pH 3–5). The flow of dye through the different compartments of the tract showed a rapid fill time for all the gut compartments and a short residence time in the crop. In all other compartments, the dye remained detectable for 10 days or longer.

Carbohydrate and amino acid metabolism in Tuber borchii mycelium during glucose utilization: a 13C NMR study

The metabolism of [1- 13C]glucose in the vegetative mycelium of the ectomycorrhizal ascomycete Tuber borchii was studied in order to characterize the biochemical pathways for the assimilation of glucose and amino acid biosynthesis. The pathways were characterized using nuclear magnetic resonance spectroscopy in conjunction with [1- 13C]glucose labeling. The enzymes of mannitol cycle and ammonium assimilation were also evaluated. The majority of the 13C label was incorporated into mannitol and this polyol was formed via a direct route from absorbed glucose. Amino acid biosynthesis was also an important sink of assimilated carbon and 13C was mainly incorporated into alanine and glutamate. From this intramolecular 13C enrichment, it is concluded that pyruvate, arising from [1- 13C]glucose catabolism, was used by alanine aminotransferase, pyruvate dehydrogenase and pyruvate carboxylase before entering the Krebs cycle. The transfer of 13C-labeled mycelium on [ 12C]glucose showed that mannitol, alanine, and glutamate carbon were used to synthesize glutamine and arginine that likely play a storage role.

Bacterial Abundance and the Activity of Microbiological Processes in the Bay of Tugur of the Sea of Okhotsk

This paper presents the results of investigation of the total abundance and the biomass of bacterioplankton, the abundance of heterotrophic bacteria, and the activity of microbiological processes involved in the carbon cycle in the water of the Bay of Tugur of the Sea of Okhotsk. In different regions of the bay, the total abundance of bacterioplankton was found to vary from 0.51 × 106 to 2.54 × 106 cells/ml; the bacterioplankton biomass, from 8.5 to 46.5 μg C/l; the abundance of heterotrophic bacteria, from 0.06 × 103 to 2.12 × 103 cells/ml; the bacterial assimilation of CO2, glucose, acetate, and protein hydrolysate, from 0.8 to 6.3, from 0.11 to 1.88, from 0.07 to 0.56, and from 0.01 to 0.22 mg C/(m3 day), respectively; the degradation of organic matter ranged from 28 to 221 mg C/(m3 day); and the intensity of methane oxidation, from 0.0005 to 0.17 μl CH4/l. The spatial pattern and the functional characteristics of bacterioplankton in the Bay of Tugur were found to be dependent on the tidal dynamics.

Role of “G-bacteria” in anaerobic substrate uptake in a SBR with no phosphorus removal

Biomass from an SBR running with no enhanced biological phosphorus removal (EBPR) but which exhibited anaerobic assimilation of glucose and acetate, was dominated by "G-bacteria", cocci in tetrads and clusters. Extracted 16S rDNA was amplified by PCR and then analysed using Denaturing Gradient Gel Electrophoresis (DGGE). Major bands were extracted and their sequences determined. Clone libraries were also prepared, the 16S rDNA extracted, PCR performed and the resultant fragments run by DGGE to aid in identifying the DGGE bands and provide fuller sequences than available by DGGE alone. The two approaches together allowed several bands to be identified. Probes for FISH analyses were designed for some of these in attempts to see to which phylogenetic group "G-bacteria" belonged, and whether they represented the dominant bands detected by DGGE. Then FISH/Microautoradiography (MAR) was used in attempts to see which bacteria there were assimilating substrates anaerobically. Results indicated that the "G-bacteria" were phylogenetically diverse, but mainly alpha-proteobacteria and members of the high G+C% gram-positive bacteria. Not all of these could assimilate glucose and/or acetate anaerobically, and Amaricoccus, the original "G-bacteria" of Cech and Hartman, was not detected.

Interactions between photoautotrophic and heterotrophic metabolism in photoheterotrophic cultures of Euglena gracilis.

Interactions between photoautotrophic and heterotrophic metabolism in photoheterotrophic culture of Euglena gracilis were studied. Under a low light supply coefficient, these two metabolic activities seem to proceed independently. The cell growth rate in photoheterotrophic culture was about the sum of the growth rates in pure photoautotrophic and heterotrophic cultures. However under a high light supply coefficient, both photoautotrophic and heterotrophic (glucose assimilation) metabolic activities were inhibited, resulting in a low photoheterotrophic growth rate. The photoheterotrophic culture was more sensitive to photoinhibition compared to the pure photoautotrophic culture. Inhibition of glucose assimilation in the photoheterotrophic culture was due to both direct and indirect (through photosynthesis) effects of high light intensity. Cell growth, glucose assimilation and alpha-tocopherol content of the cells were higher when ambient air was used for aeration than when a mixture of carbon dioxide and air was used. Even when photosynthesis was inhibited by addition of 3-(3,4-dichlorophenyl)- 1,1-dimethylurea to photoheterotrophic culture, light stimulated alpha-tocopherol synthesis by E. gracilis.

Continuous cultivation of the diatom Nitzschia laevis for eicosapentaenoic acid production: physiological study and process optimization.

The continuous cultures of the diatom Nitzschia laevis were performed at different dilution rates (D) and feed glucose concentrations (S(0)) to investigate cellular physiological responses and its production potential of eicosapentaenoic acid (EPA). Steady-state cell dry weight, residual glucose concentration, cell growth yield, specific glucose consumption rate, and fatty acid profiles were investigated within the range of D from 0.1 to 1.0 day(-1) (S(0) fixed at 20 g/L) and the range of S(0) from 5 to 35 g/L (D fixed at 0.3 day(-1)), respectively. The highest EPA productivity of 73 mg L(-1) day(-1) was obtained at D = 0.5 day(-1) and S(0) = 20 g/L. However, when the continuous culture achieved high productivities of EPA at certain dilution rates and feed glucose concentrations, glucose in the feed could not be consumed completely. Accordingly, the continuous culture was evaluated in terms of both EPA productivity (P) and glucose assimilation efficiency (E). The parameter eta, defined as the product of P and E, was used as an overall performance index. Since eta is a function of the two independent variables D and S(0), we employed a central composite design to optimize D and S(0) for the highest eta value. Based on the experimental results of the design, a second-order polynomial equation was established to represent the relationship between eta and D and S(0). The optimal values of D and S(0) were subsequently determined as 0.481 day(-1) and 15.56 g/L, respectively by the empirical model. The verification experiment confirmed the validity of the model. Under the optimal conditions, eta value reached 46.5 mg L(-1) day(-1), suggesting a considerably high efficiency of the continuous culture of N. laevis in terms of EPA production and glucose utilization.

Methods for assessing intestinal absorptive function in relation to enteral nutrition.

The success of nasoenteral nutrition support can be limited by intestinal impairment. In particular, reduced absorptive area, mucosal atrophy and abnormal motility may reduce absorption of macronutrients and micronutrients, and diarrhoea remains a commonly encountered complication. We review how basic physiological techniques can be used to investigate such pathophysiology. Lumenal nutrients control mucosal growth, expression of mucosal transporters and regional gut motility. Cell biology techniques now complement classical intestinal perfusion methods in determining the 'safety factor' of excess absorptive capacity. The controversial role of the sodium-glucose linked transporter in dietary glucose assimilation is described in terms of its control, its true function and its role in uptake of other solutes. Techniques that involve brush-border membrane vesicles, Caco-2 cells, mucosal immunohistochemistry and gene expression probes are described. Together, these techniques describe a picture of an organ with remarkable ability to maintain digestive and absorptive function in response to a wide variety of nutritional intakes, often in the face of inflammatory illness.

A deeper investigation on carbohydrate cycling in Sinorhizobium meliloti

Recycling of triose-phosphate and pentose-phosphate was previously reported on glucose in Sinorhizobium meliloti, a polysaccharide-synthesizing bacterium, but the metabolic basis of such processes remained unclear. In this work, we have used 13C-labelling strategies to demonstrate that carbohydrate cycling in this organism is independent of the gluconate bypass, the alternative pathway for glucose assimilation involving its periplasmic oxidation into gluconate. Furthermore, carbohydrate cycling in S. meliloti is also observed on fructose, making the situation in this bacterium significantly different from that depicted for alginate-synthesizing species.

Selection and characterization of mixed starter cultures for lactic acid fermentation of carrot, cabbage, beet and onion vegetable mixtures

An evaluation of various lactic acid bacteria (LAB) for the fermentation of cabbage, carrot and beet-based vegetable products was carried out. As part of a screening process, the growth of 15 cultures in a vegetable juice medium (VJM) was characterized by automated spectrophotometry. Acidification patterns as well as viability during storage of the LAB were also established. There were greater differences between the pure cultures than the mixed ones with respect to growth in VJM and viability during storage. Reductions in viable cell counts during storage of the fermented VJM occurred more rapidly with a Leuconostoc strain than for pediococci or lactobacilli. Inoculation of vegetables was carried out with cultures of Lactobacillus plantarum NK-312, Pediococcus acidilactici AFERM 772 and Leuconostoc mesenteroides BLAC which were rehydrated in a brine. This rehydration procedure was not detrimental to viability. During fermentation of a carrot/cabbage vegetable mix, sugar metabolism was characterized by the assimilation of both glucose and fructose, but sugars remained in the fermented vegetables when acidification stopped. The pH in the LAB-inoculated vegetables after 72 h at 20°C was significantly lower (by 0.2 units) than the uninoculated control. Inoculation with LAB designed for silage fermentation resulted in the inhibition of acetic acid production, and reduced the production of ethanol during fermentation. The selection process on VJM enabled the preparation of a mixed culture that was more rapid than the silage inoculants in acidifying the medium and was more effective in reducing the production of gas during the fermentation and storage of the fermented vegetables.

Structure and Functional Analysis of the Microbial Community in an Aerobic: Anaerobic Sequencing Batch Reactor (SBR) with no Phosphorus Removal

The bacterial community of an aerobic:anaerobic non-P removing SBR biomass fed a mixture of acetate and glucose was analysed using several 16S rRNA based methods. Populations responsible for anaerobic glucose and acetate assimilation were determined with fluorescent in situ hybridization (FISH) in combination with microautoradiography (FISH/MAR). At 'steady state' this community consisted of alpha-Proteobacteria (26%) and gamma-Proteobacteria (14%), mainly appearing as large cocci in tetrads (i.e. typical 'G-Bacteria'). Large numbers of low G+C bacteria (22%), and high G+C Gram-positive bacteria (29%) seen as small cocci in clusters or in sheets were also detected after FISH. DGGE fingerprinting of PCR amplified 16S rDNA fragments and subsequent cloning and sequencing of several of the major bands led to the identification of some of these populations. They included an organism 98% similar in its 16S rRNA sequence to Micropruina glycogenica, and ca. 76% of the high G+C bacteria responded to a probe MIC 184, designed against it. The rest responded to the KSB 531 probe designed against a high G+C clone sequence, sbr-gs28 reported in other similar systems. FISH analyses showed that both these high G+C populations were almost totally dominated by small clustered cocci. Only ca. 2% of cells were beta-Proteobacteria. None of the alpha- and gamma-Proteobacterial 'G-bacteria' responded to FISH probes designed for the 'G-Bacteria' Amaricoccus spp. or Defluvicoccus vanus. FISH/MAR revealed that not all the alpha-Proteobacterial 'G-Bacteria' could take up acetate or glucose anaerobically. Almost all of the gamma-Proteobacterial 'G-Bacteria' assimilated acetate anaerobically but not glucose, the low G+C clustered cocci only took up glucose, whereas the high G+C bacteria including M. glycogenica and the sbr-gs28 clone assimilated both acetate and glucose. All bacteria other than the low G+C small cocci and a few of the alpha-Proteobacteria accumulated PHB. The low G+C bacteria showing anaerobic glucose assimilation ability were considered responsible for the lactic acid produced anaerobically by this SBR biomass, and M. glycogenica for its high glycogen content.

Cytochemical and physiological studies of the energetic metabolism and osmotrophy in Sagitta friderici (chaetognath)

Different glucidic labelled compounds were used for carbohydrate metabolism exploration, tissular glycogen storage localization and amino acid uptake response to osmotic variation in the adult of Sagitta friderici.After [1-14C] glucose or [6-14C] glucose (osmotrophic) assimilation, the relative activity of the different metabolic pathways (glycolysis–citric acid cycle and pentose cycle) have been estimated by means of a micro-radiorespirometric method. The kinetics of expired 14CO2 showed that adult Sagitta mainly used the glycolytic–Krebs cycle rather than the pentose cycle in glucose catabolism.After 2-deoxy-D-[1-14C] glucose (osmotrophic) assimilation, tissular glycogen storages were localized. It was shown that glycogen deposits are abundant in several tissues except the locomotor muscles, which probably represent the main glucose consumers in these very active animals. The problem of glucose transport from the storage localizations to the sites of utilization has been discussed.The L-3H-leucine uptake variations into the Sagitta body after an osmotic shock have been estimated.

Anomalies in the growth kinetics of Saccharomyces cerevisiae strains in aerobic chemostat cultures

Aerobic glucose-limited chemostat cultivations were conducted with Saccharomyces cerevisiae strains NRRL Y132, ATCC 4126 and CBS 8066, using a complex medium. At low dilution rates all three strains utilised glucose oxidatively with high biomass yield coefficients, no ethanol production and very low steady-state residual glucose concentrations in the culture. Above a threshold dilution rate, respiro-fermentative (oxido-reductive) metabolism commenced, with simultaneous respiration and fermentation occurring, which is typical of Crabtree-positive yeasts. However, at high dilution rates the three strains responded differently. At high dilution rates S. cerevisiae CBS 8066 produced 7–8 g ethanol L−1 from 20 g glucose L−1 with concomitant low levels of residual glucose, which increased markedly only close to the wash-out dilution rate. By contrast, in the respiro-fermentative region both S. cerevisiae ATCC 4126 and NRRL Y132 produced much lower levels of ethanol (3–4 g L−1) than S. cerevisiae CBS 8066, concomitant with very high residual sugar concentrations, which was a significant deviation from Monod kinetics and appeared to be associated either with high growth rates or with a fermentative (or respiro-fermentative) metabolism. Supplementation of the cultures with inorganic or organic nutrients failed to improve ethanol production or glucose assimilation. Journal of Industrial Microbiology & Biotechnology (2000) 24, 231–236.

The Effect of Vitamins and Amino Acids on Glucose Uptake in Aerobic Chemostat Cultures of three Saccharomyces cerevisiae Strains

In the respiro-fermentative region of aerobic chemostat cultures at steady state, Saccharomyces cerevisiae CBS 8066 produced high concentrations of ethanol with concomitant low levels of residual glucose which followed Monod kinetics. By contrast, very high residual glucose concentrations were observed in cultures of S. cerevisiae strains ATCC 4126 and NRRL Y132 at dilution rates above 60% of the washout dilution rate, resulting in much lower ethanol concentrations, even though clearly glucose-limited at lower dilution rates in the respiratory region. The addition of a vitamin mixture resulted in decreased residual glucose concentrations in respiro-fermentative cultures of all three strains, but the effect was much more pronounced with strains ATCC 4126 and NRRL Y132. Meso-inositol was mainly responsible for this effect, although with strain ATCC 4126 other vitamins as well as an amino acid mixture were also required to minimise the steady-state residual glucose levels. The residual glucose concentration in continuous culture was, therefore, greatly dependent on the growth factor requirements of the particular yeast strain, which apparently increased on increasing the dilution rate into the respiro-fermentative region. The strain differences with respect to growth factor requirements at high dilution rates, which were not evident at low dilution rates, had a profound effect on the kinetics of glucose assimilation in aerobic chemostat culture.

Bacterial utilization of dissolved glucose in the upper water column of the Gulf of Mexico

Several aspects of bacterial glucose assimilation and bacterial production (BP) were investigated over 2 d in the upper 300 m of the Gulf of Mexico. Glucose concentrations ranged from 2 to 15 nM in surface seawater; glucose utilization rates were 1–3 nM d−1 in the upper 150 m of the water column and 0.02–0.8 nM d−1 in deeper water (150–300 m). Turnover of glucose and bacteria were similar and were in the range of 0.01–0.4 d−1. Measured glucose assimilation rates imply that glucose supports ∼5–10% of the observed BP in surface waters. The limiting factors for bacterial glucose utilization varied with depth. Glucose assimilation seemed to be limited by availability of inorganic N in the chlorophyll maximum. Additions of nitrate or ammonium had similar effects on glucose assimilation. BP was limited by C below and above the chlorophyll maximum. Additions of high‐molecular‐weight (HMW) dissolved organic matter (DOM) decreased bacterial glucose uptake rates, indicating that substances competing with glucose assimilation were derived from HMW DOM extracellularly. Thus, the measurement of free glucose assimilation appears to integrate bacterial utilization of combined glucose and indicates that many of the free sugars in seawater may be derived from polymers. The higher glucose assimilation rates achieved with inorganic nutrient additions indicate that bacterial degradation of C‐rich organic matter (OM) could vary between ocean regions with varying inorganic nutrient concentrations.

A Zymomonas mobilis mutant with delayed growth on high glucose concentrations.

Exponentially growing cells of Zymomonas mobilis normally exhibit a lag period of up to 3 h when transferred from 0.11 M (2%) to 0.55 M (10%) glucose liquid medium. A mutant of Z. mobilis (CU1Rif2), fortuitously isolated, showed more than a 20-h lag period when grown under the same conditions, whereas on 0.55 M glucose solid medium, it failed to grow. The growth of CU1Rif2 on elevated concentrations of other fermentable (0.55 M sucrose or fructose) or nonfermentable (0.11 M glucose plus 0.44 M maltose or xylose) sugars appeared to be normal. Surprisingly, CU1Rif2 cells grew without any delay on 0.55 M glucose on which wild-type cells had been incubated for 3 h and removed at the beginning of their exponential phase. This apparent preconditioning was not observed with medium obtained from wild-type cells grown on 0.11 M glucose and supplemented to 0.55 M after removal of the wild-type cells. Undelayed growth of CU1Rif2 on 0.55 M glucose previously conditioned by the wild type was impaired by heating or protease treatment. It is suggested that in Z. mobilis, a diffusible proteinaceous heat-labile factor, transitionally not present in 0.55 M glucose CU1Rif2 cultures, triggers growth on 0.55 M glucose. Biochemical analysis of glucose uptake and glycolytic enzymes implied that glucose assimilation was not directly involved in the phenomenon. By use of a wild-type Z. mobilis genomic library, a 4.5-kb DNA fragment which complemented in low copy number the glucose-defective phenotype as well as glucokinase and glucose uptake of CU1Rif2 was isolated. This fragment carries a gene cluster consisting of four putative coding regions, encoding 167, 167, 145, and 220 amino acids with typical Z. mobilis codon usage, -35 and -10 promoter elements, and individual Shine-Dalgarno consensus sites. However, strong homologies were not detected in a BLAST2 (EMBL-Heidelberg) computer search with known protein sequences.

Test, rejection, and reformulation of a chemical reactor-based model of gut function in a fruit-eating bird.

We explored modulation of retention time in cedar waxwings (Bombycilla cedrorum) by feeding them diets varying in hexose concentration. Our goals were to (1) test three predictions of a chemical reactor-based model of how guts might respond optimally to diet shifts; (2) determine whether modulation of retention time can occur quickly, thereby facilitating rapid changes in diet; (3) tease apart the relative influence of ingestion rate and nutrient concentration on retention time; and (4) examine the degree of axial mixing in the intestine and its relationship with retention time. The model's predictions were rejected: mean retention time did not decrease, ingestion rate did not increase, and glucose assimilation efficiency did not decrease with increased hexose concentration of the diet. Instead, birds displayed maximal intake rate at intermediate sugar concentration, and mouth to cloaca mean retention times increased with hexose concentration. Significant modulation of retention time occurred quickly, within 3 h of exposure to a different diet. Birds did equally well in terms of total energy assimilated on diets differing 3.3-fold in hexose concentration (from 500 mmol/L to 1660 mmol/L) but showed reduced intake when fed food with low hexose concentration (110 mmol/L). Far more variation in retention time was explained by direct effects of ingestion rate than by direct effects of hexose concentration. Finally, a gut dispersion index that measured degree of axial mixing was positively correlated with mean retention time, indicating that higher retention times are accompanied by increased axial mixing. We propose a modification of the assumptions of the original model. The resulting "osmotic constraint" model better captures the interaction between feeding rate and digestive function in fruit-eating birds.

Cardiovascular disease and risk factors in adults with hypopituitarism.

In 1992, Cuneo et al. (1992) reviewed, in this journal, the consequences of growth (GH) deficiency during adult life and coined the term ‘Growth hormone deficiency (GHD) syndrome in adults’. Limited information was available then on cardiovascular disease and risk factors in adults with hypopituitarism. More data have been published since on cardiovascular morbidity in symptom-free hypopituitary adults. Several groups have also published results on the effects of GH therapy on cardiac structure and function and on the cardiovascular risk factors. We shall review here the epidemiological and experimental data on cardiovascular morbidity and mortality in adult hypopituitarism. We shall also examine the prevalence of cardiovascular risk factors in this group of patients and their potential causes focusing on the possible role of GH deficiency and the effects of GH replacement.

Detection and identification of wild yeasts in lager breweries

Wild yeasts were detected in 41 out of 101 brewery yeast samples investigated using six different selective principles. Malt extract, yeast extract, glucose, peptone (MYGP) agar supplemented with 195 ppm CuSO 4 was found to be the most effective selective principle, detecting wild yeasts in 80% of the contaminated samples. Both Saccharomyces and non- Saccharomyces wild yeasts were detected on this medium. Lysine medium, crystal violet medium and incubation of non-selective media at 37°C detected wild yeasts in 46–56% of the contaminated samples. On using actidione medium, only 20% of the wild yeasts were detected. The combined use of MYGP supplemented with 195 ppm CuSO 4 and one of the other selective principles did not improve the recovery of the wild yeasts. The wild yeasts found consisted of Saccharomyces cerevisiae (57%), Pichia spp. (28%) and Candida spp. (15%). Using the API ID 32 C kit, 35 different assimilation profiles were obtained for the 124 wild yeast isolates investigated. All isolates were capable of glucose assimilation, whereas only 79% of the isolates assimilated saccharose, 75% maltose, 70% galactose, 65% raffinose and 65% lactate. Lactose, inositol, rhamnose and glucuronate were not assimilated by any of the isolates. The differences in assimilation pattern did not reflect any differences in recovery by the selective principles investigated. The majority of the wild yeast isolates investigated were capable of growth in wort and beer, indicating their possible role as spoilage organisms. The Sacch. cerevisiae isolates were found to be the most hazardous, with some isolates being capable of extensive growth in bottled beer within seventeen days at ambient temperature.