A Combination of Essential Fatty Acids, Panax Ginseng Extract, and Green Tea Catechins Modifies Brain fMRI Signals in Healthy Older Adults.

OBJECTIVES: To assess the effects of a combination of omega 3 essential fatty acids, green tea catechins, and ginsenosides on cognition and brain functioning in healthy older adults. DESIGN: Double-blind, placebo-controlled, crossover design randomized controlled trial with 26-day intervention phases and a 30-day washout period. SETTING: The Institute for Dementia Research and Prevention at the Pennington Biomedical Research Center. PARTICIPANTS: Ten independently-living, cognitively-healthy older adults (mean age: 67.3 + 2.01 years). INTERVENTION: Daily consumption of an investigational product (trade name "Cerbella TM") consisting of an emulsified liquid combination of standardized fish oil, panax ginseng extract, and green tea catechins in a flavored base of lecithin phospholipids optimized to maximize bioavailability of the active ingredients. MEASUREMENTS: Before and after supplementation with the investigational product or placebo, participants completed cognitive tests including the Mini Mental State Exam (MMSE), Stroop test, Digit Symbol Substitution Test (DSST), and Immediate and Delayed Recall tests, as well as functional magnetic resonance imaging (fMRI) during a standard cognitive task switching paradigm. RESULTS: Performance on the MMSE, Stroop test, and DSST increased significantly over one month of supplementation with the investigational product (one-sample t tests, p<.05) although differences between these changes and corresponding changes during supplementation with placebo were not significant (two-sample t tests, p>.05). During supplementation with the investigational product, brain activation during task performance increased significantly more than during supplementation with placebo in brain regions known to be activated by this task (anterior and posterior cingulate cortex). Functional connectivity during task execution between task regions (middle frontal gyrus and anterior cingulate cortex) increased significantly during supplementation with the investigational product, relative to placebo. Functional connectivity during rest between task regions (precentral gyrus and middle frontal gyrus) and default mode network regions (medial frontal gyrus and precuneus) decreased during supplementation with the investigational product relative to placebo, suggesting greater segregation of task and rest related brain activity. CONCLUSION: One-month supplementation with a combination of omega 3 essential fatty acids, green tea catechins, and ginsenosides was associated with suggestive changes in cognitive functioning as well as modification of brain activation and brain functional connectivity in cognitively healthy older adults.

[Acute colonic pseudo-obstruction: Ogilvie syndrome]. / Akute Kolonpseudoobstruktion: Ogilvie-Syndrom.

Acute colonic pseudo-obstruction (ACPO) is characterized by marked colonic dilatation which develops over several days. ACPO is due to a motility disorder and is not caused by colonic obstruction and occurs in patients with severe, often acute underlying diseases or postoperatively. It is associated with a 25-30% mortality overall that increases to up to 50% in patients who develop complications (e.g. colonic ischemia and perforation). The pathogenesis of the disorder has not yet been clarified and clinical symptoms and signs are relatively unspecific. In particular, ACPO has to be differentiated from colonic obstruction and toxic megacolon. For this blood tests and radiological tests are required, e.g. plain abdominal radiograph, abdominal computed tomography (CT) and water soluble contrast enema, which are also required for detection of complications. Patients with ACPO should generally receive supportive therapy for decompression of the gastrointestinal tract (e.g. gastric and rectal tubes) and to minimize predisposing factors. In most uncomplicated cases this leads to resolution of colonic dilatation. Clinical and radiological controls at close intervals are required until the condition is resolved. If patients do not respond within 1-2 days or if ACPO has already reached a critical duration (>3-4 days) or extent (i.e. cecal diameter ≥12 cm), neostigmine should be administered and leads to durable success in approximately 3 out of 4 patients. Patients who are still refractory to treatment should receive endoscopic decompression. More invasive therapeutic options, such as cecostomy or (segmental) colonic resection should only be considered for patients who still do not respond to treatment or present with the abovementioned complications.

Calcium gluconate supplementation is effective to balance calcium homeostasis in patients with gastrectomy.

UNLABELLED: We demonstrate histological evidence for hyperparathyroidism in patients with gastrectomy. This is, at least in part, explained by impaired calcium absorption, resulting in mineralization defects and secondary hyperparathyroidism. Additionally, we demonstrate improved bone mineralization in patients with gastrectomy after gluconate therapy and showed the effectiveness of calcium gluconate over carbonate to balance impaired calcium hemostasis in mice. INTRODUCTION: Gastrectomy and hypochlorhydria due to long-term proton pump inhibitor therapy are associated with increased fracture risk because of intestinal calcium malabsorption. Hence, our objectives were to histologically investigate bone metabolism in patients with gastrectomy and to analyze the impact of calcium gluconate supplementation on skeletal integrity in the setting of impaired gastric acidification. METHODS: Undecalcified bone biopsies of 26 gastrectomized individuals were histologically analyzed. In the clinical setting, we retrospectively identified 5 gastrectomized patients with sufficient vitamin D level, who were additionally supplemented with calcium gluconate and had a real bone mineral density (aBMD) follow-up assessments. A mouse model of achlorhydria (ATP4b-/-) was used to compare the effect of calcium gluconate and calcium carbonate supplementation on bone metabolism. RESULTS: Biopsies from gastrectomized individuals showed significantly increased osteoid, osteoclast, and osteoblast indices and fibroosteoclasia (p < 0.05) as well as impaired calcium distribution in mineralized bone matrix compared to healthy controls. Five gastrectomized patients with sufficient vitamin D level demonstrated a significant increase in aBMD after a treatment with calcium gluconate alone for at least 6 months (p < 0.05). Calcium gluconate was superior to calcium carbonate in maintaining calcium metabolism in a mouse model of achlorhydria. CONCLUSION: Gastrectomy is associated with severe osteomalacia, marrow fibrosis, and impaired calcium distribution within the mineralized matrix. We show that calcium gluconate supplementation can increase bone mineral density in gastrectomized individuals and performs superior to calcium carbonate in restoring calcium/skeletal homoeostasis in a mouse model of achlorhydria.

Platforms for energy and nutrient recovery from domestic wastewater: A review.

Alternative domestic wastewater treatment processes that recover energy and nutrients while achieving acceptable nutrient limits (<5mgNL(-1)) are a key challenge. Major drivers are value and availability of phosphorous, nitrogen, and potassium, and increasing energy costs. The two major platforms that can achieve this are (a) low energy mainline (LEM), with low strength anaerobic treatment, followed by mainline anaerobic nitrogen removal and chemical or adsorptive phosphorous removal and (b) partition-release-recover (PRR), in which carbon and nutrients are partitioned to solids through either heterotrophic or phototrophic microbes, followed by anaerobic digestion of these solids and recovery from the digestate. This paper reviews practical application of these processes, with a focus on energy costs. Compared to conventional processes which require 0.5kWhkL(-1) electricity (500mgCODL(-1) influent concentration), PRR requires only 0.05kWhkL(-1) electricity. LEM offers the possibility to recover 0.1kWhkL(-1) as electricity with net energy generation above 400mgCODL(-1)influent, while PRR becomes energy generating at >650mgCODL(-1). PRR offers the possibility for recovery of nitrogen and other nutrients (including potassium) through assimilative recovery. However, the energetic overhead of this is substantial, requiring 5kWhkgN(-1) as electricity, which compares to ammonia fixation costs. The lower energy costs, and near to market status of LEM treatment make it likely as a recovery platform in the shorter term, while ability to recover other elements such as nitrogen and potassium, as well as enhance favourability on concentrated wastewaters may enhance the desirability of partitioning in the longer term.

Understanding colloidal FeSx formation from iron phosphate precipitation sludge for optimal phosphorus recovery.

The use of sulfide to form iron sulfide precipitates is an attractive option for separation and recovery of phosphorus and ferric iron from ferric phosphate sludge generated in wastewater treatment. The key factors affecting the simultaneous generation and separation of iron sulfide precipitates and phosphate solution from ferric phosphate sludge have so far not been thoroughly investigated. This study therefore focuses on the recovery of phosphorus from synthetic sludge by controlled sulfide addition under different operating conditions. The factors that affect the phosphorus recovery, as well as the optimal process conditions to achieve an effective solid-liquid separation, were investigated. The separation of the FeSx particles is a significant challenge due to the colloidal nature of the particles formed. Faster separation and higher phosphorus recovery was achieved when operating at pH 4 with dosing times of at least 1h. At this pH, phosphorus recovery of 70±6% was reached at the stoichiometric S/Fe molar ratio of 1.5, increasing to over 90% recovery at a S/Fe molar ratio of 2.5. Zeta potential results confirmed the colloidal nature of the iron sulfide precipitate, with the isoelectric point around pH 4, explaining the fast separation of the FeSx particles at this pH.

Effects of fish oil and conjugated linoleic acids on carnitine homeostasis in laying hens.

1. The effects of n-3 polyunsaturated fatty acids (PUFA) and conjugated linoleic acids (CLA) on genes involved in carnitine homeostasis were compared in laying hens. Three groups of laying hens were fed on a control diet or a diet with either 3% of fish oil or CLA for 4 weeks. 2. Feed intake and egg production rate did not differ between the three groups. Diets with fish oil or CLA had only a weak effect on mRNA levels of PPARα target genes (ACO, CPT-I) in the liver and did not influence mRNA concentrations of the most important carnitine transporter OCTN2, enzymes of involved in carnitine synthesis (TMLD, TMABA-DH, BBD) or concentrations of carnitine in plasma, liver and total egg contents. 3. Hens fed the CLA diet had lower concentrations of free and total carnitine in egg yolk but higher concentrations of carnitine in albumen than control hens (P < 0·05), whereas the amount of free and total carnitine in whole egg did not differ. 4. In conclusion, the study showed that feeding fish oil or CLA causes only a weak activation of PPARα in tissues of laying hens that probably explained the lack of effect on carnitine homeostasis. The results contrast with those in humans and mice that show a significant effect of synthetic PPARα agonists on carnitine homeostasis in humans and mice.

Supplementation with l-carnitine downregulates genes of the ubiquitin proteasome system in the skeletal muscle and liver of piglets.

Supplementation of carnitine has been shown to improve performance characteristics such as protein accretion in growing pigs. The molecular mechanisms underlying this phenomenon are largely unknown. Based on recent results from DNA microchip analysis, we hypothesized that carnitine supplementation leads to a downregulation of genes of the ubiquitin proteasome system (UPS). The UPS is the most important system for protein breakdown in tissues, which in turn could be an explanation for increased protein accretion. To test this hypothesis, we fed sixteen male, four-week-old piglets either a control diet or the same diet supplemented with carnitine and determined the expression of several genes involved in the UPS in the liver and skeletal muscle. To further determine whether the effects of carnitine on the expression of genes of the UPS are mediated directly or indirectly, we also investigated the effect of carnitine on the expression of genes of the UPS in cultured C2C12 myotubes and HepG2 liver cells. In the liver of piglets fed the carnitine-supplemented diet, the relative mRNA levels of atrogin-1, E214k and Psma1 were lower than in those of the control piglets (P < 0.05). In skeletal muscle, the relative mRNA levels of atrogin-1, MuRF1, E214k, Psma1 and ubiquitin were lower in piglets fed the carnitine-supplemented diet than that in control piglets (P < 0.05). Incubating C2C12 myotubes and HepG2 liver cells with increasing concentrations of carnitine had no effect on basal and/or hydrocortisone-stimulated mRNA levels of genes of the UPS. In conclusion, this study shows that dietary carnitine decreases the transcript levels of several genes involved in the UPS in skeletal muscle and liver of piglets, whereas carnitine has no effect on the transcript levels of these genes in cultivated HepG2 liver cells and C2C12 myotubes. These data suggest that the inhibitory effect of carnitine on the expression of genes of the UPS is mediated indirectly, probably via modulating the release of inhibitors of the UPS such as IGF-1. The inhibitory effect of carnitine on the expression of genes of the UPS might explain, at least partially, the increased protein accretion in piglets supplemented with carnitine.

Inhibition of the pro-inflammatory NF-κB pathway by a grape seed and grape marc meal extract in intestinal epithelial cells.

In pigs and other monogastric animal, the weaning phase is commonly accompanied by an increased susceptibility to gut disorders such as diarrhoea owing to the induction of an inflammatory process in the intestine during weaning. Given the unfavourable effects of intestinal inflammation on feed consumption, digestive capacity of the intestine and growth of animals, controlling intestinal inflammation is a reasonable approach for the maintenance of performance characteristics of livestock animals. Therefore, this study aimed to study the anti-inflammatory potential of a commercial polyphenol-rich grape seed (GS) and grape marc (GM) meal-based feed additive in a well-established in vitro intestinal epithelium model (polarized Caco-2 cells). The anti-inflammatory potential was evaluated by studying the effect of an ethanolic extract obtained from the GS and GM meal-based feed additive (GSGME) on the pro-inflammatory transcription factor NF-κB, which is considered to play a key role in the induction of weaning-associated intestinal inflammation. The highest non-cytotoxic concentrations of the ethanolic GSGME dose dependently reduced TNFα-induced NF-κB transactivation and decreased TNFα-induced mRNA levels of the NF-κB target genes IL-1ß, IL-8, MCP-1 and CXCL1 in Caco-2 intestinal cells (p < 0.05). No effect of the ethanolic GSGME was observed on the cytoprotective Nrf2 pathway in Caco-2 cells as evidenced by an unaltered Nrf2 transactivation and unchanged mRNA levels of Nrf2 target genes, such as GPX-2, NQO1, CYP1A1 and UGT1A1. In conclusion, this study shows that an ethanolic GSGME exerts anti-inflammatory effects in intestinal cells under in vitro conditions. Thus, polyphenol-rich GSGM meal-based feed additives may be useful for the inhibition or prevention of inflammatory processes in the intestine of livestock animals, in particular during states with inappropriate NF-κB activation in the intestinal tissue, such as the weaning phase. Future studies are warranted to prove the in vivo anti-inflammatory potential of GSGM meal-based feed additives.

Struggling with recurrent Clostridium difficile infections: is donor faeces the solution?

Patients with recurrent Clostridium difficile infections (CDI) in hospitals and the community constitute an increasing treatment problem. While most patients with a first infection respond to either metronidazole or oral vancomycin, therapy in recurrent C. difficile infections tends to fail repeatedly. Lack of alternative treatment options can be a tremendous burden, both to patients and their treating physicians. Most guidelines recommend prolonged oral vancomycin pulse and or tapering schedules, but evidence-based treatment strategies are lacking. The role of immunoglobulins, whey prepared from vaccinated cows, probiotics or other antibiotics is unclear. Since 1958 several case series and case reports describe a treatment strategy where faecal infusions are successfully given for the treatment of recurrent CDI. Restoring intestinal flora has been historically thought of as the mechanism responsible for cure in these patients. In the literature, more than 150 patients have received faeces from a healthy donor, either infused through an enema, or through a nasoduodenal or nasogastric tube. We summarise the literature regarding treatment with donor faeces for recurrent CDI, and introduce the FECAL trial, currently open for inclusion.

Two-stage thermophilic-mesophilic anaerobic digestion of waste activated sludge from a biological nutrient removal plant.

A two-stage thermophilic-mesophilic anaerobic digestion pilot-plant was operated solely on waste activated sludge (WAS) from a biological nutrient removal (BNR) plant. The first-stage thermophilic reactor (HRT 2 days) was operated at 47, 54 and 60 degrees C. The second-stage mesophilic digester (HRT 15 days) was held at a constant temperature of 36-37 degrees C. For comparison with a single-stage mesophilic process, the mesophilic digester was also operated separately with an HRT of 17 days and temperature of 36-37 degrees C. The results showed a truly thermophilic stage (60 degrees C) was essential to achieve good WAS degradation. The lower thermophilic temperatures examined did not offer advantages over single-stage mesophilic treatment in terms of COD and VS removal. At a thermophilic temperature of 60 degrees C, the plant achieved 35% VS reduction, representing a 46% increase compared to the single-stage mesophilic digester. This is a significant level of degradation which could make such a process viable in situations where there is no primary sludge generated. The fate of the biologically stored phosphorus in this BNR sludge was also investigated. Over 80% of the incoming phosphorus remained bound up with the solids and was not released into solution during the WAS digestion. Therefore only a small fraction of phosphorus would be recycled to the main treatment plant with the dewatering stream.

Effects of solids concentration, pH and carbon addition on the production rate and composition of volatile fatty acids in prefermenters using primary sewage sludge.

Increasing evidence is emerging that the performance of enhanced biological phosphorus removal (EBPR) systems relies on not only the total amount but also the composition of volatile fatty acids (VFAs). Domestic wastewater often contains limited amounts of VFAs with acetic acid typically being the dominating species. Consequently, prefermenters are often employed to generate additional VFAs to meet the demand for carbon by EBPR and/or denitrification processes. Limited knowledge is currently available on the effects of operational conditions on the production rate and composition of VFAs in prefermenters. In this study, a series of controlled batch experiments were conducted with sludge from a full-scale prefermenter to determine the impact of solids concentration, pH and addition of molasses on prefermentation processes. It was found that an increase in solids concentration enhanced total VFA production with an increased propionic acid fraction. The optimal pH for prefermentation was in the range of 6-7 with significant productivity loss when pH was below 5.5. Molasses addition significantly increased the production of VFAs particularly the propionic acid. However, the fermentation rate was likely limited by the biological activity of the sludge rather than by the amount of molasses added.

Modelling biological processes under anaerobic conditions through integrating titrimetric and off-gas measurements-applied to EBPR systems.

An innovative method for modelling biological processes under anaerobic conditions is presented and discussed. The method is based on titrimetric and off-gas measurements. Titrimetric data is recorded as the addition rate of hydroxyl ions or protons that is required to maintain pH in a bioreactor at a constant level. An off-gas analysis arrangement measures, among other things, the transfer rate of carbon dioxide. The integration of these signals results in a continuous signal which is solely related to the biological reactions. When coupled with a mathematical model of the biological reactions, the signal allows a detailed characterisation of these reactions, which would otherwise be difficult to achieve. Two applications of the method to the enhanced biological phosphorus removal processes are presented and discussed to demonstrate the principle and effectiveness of the method.

Short-term effects of carbon source on the competition of polyphosphate accumulating organisms and glycogen accumulating organisms.

The effectiveness of enhanced biological phosphorus removal (EBPR) systems is directly affected by the competition of polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigated the short-term effects of carbon source on PAO and GAO performance. The tests were designed to clearly determine the impact of volatile fatty acid (VFA) composition on the performance of two types of biomass, one enriched for PAOs and the other for GAOs. The two populations were enriched in separate reactors using identical operating conditions and very similar influent compositions with acetate as the sole carbon source. The only difference was that a very low level of phosphorus was present in the influent to the GAO reactor. The abundance of PAOs and GAOs was quantified using fluorescence in-situ hybridisation. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to utilise different carbon substrates. While both are able to take up acetate rapidly and completely, the GAOs are far slower at consuming propionate than the PAOs during short-term substrate changes. This provides a potentially highly valuable avenue to influence the competition between PAOs and GAOs. Other VFAs studied seem to be less usable in the short term by both PAOs and GAOs, as indicated by their much lower uptake rates.

A novel wastewater treatment process: simultaneous nitrification, denitrification and phosphorus removal.

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen concentration (DO, 0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHA), accompanied with phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to less than 0.5 mg/L at the end of the cycle. Ammonia was also oxidised during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis found that the final denitrification product was mainly nitrous oxide (N2O) not N2. Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms rather than denitrifying polyphosphate-accumulating organisms were responsible for the denitrification activity.

Optimisation of Noosa BNR plant to improve performance and reduce operating costs.

Noosa WWTP is publicly owned and privately operated by Australian Water Services. The process includes primary sedimentation, raw sludge fermentation, biological nutrient removal (BNR), sand filtration and ultraviolet (UV) disinfection. An innovative feature of the plant is the supplementary carbon dosing facility to avoid the use of metal salts (alum or ferric) for phosphorus removal. The average flow treated during 2000 was 9.0 ML/d. The annual 50 percentile effluent quality requirements for nutrients are total N < 5 mg/L and total P < 1 mg/L. The objectives of this project were to: determine the cause of variability in phosphorus removal; develop a strategy to control the variability in phosphorus removal; and minimise the operating cost of supplementary carbon dosing while achieving the effluent quality requirements. An investigation of chemical and microbiological parameters was implemented and it was concluded that there were several factors causing variability in phosphorus removal, rather than a single cause. The following four major causes were identified, and the control strategies that were adopted resulted in the plant achieving annual 50 percentile effluent total P = 0.37 mg/L and total N = 3.0 mg/L during 2001. First, phosphorus removal was limited by the available VFA supply due to consumption of VFA by other organisms competing with phosphate accumulating organisms (PAO), and due to diurnal variations in the sewage VFA and phosphate concentrations. Therefore, supplementary carbon dosing was essential to make allowance for competing reactions. Second, increasing the fermenter VFA yield via supplementary carbon dosing with molasses was found to be an effective and economic way of ensuring reliable phosphorus removal. Third, nitrate in the RAS resulted in consumption of VFA by denitrifying bacteria, particularly with process configurations where the RAS was recycled directly into the anaerobic zone. Incorporating a RAS denitrification zone into the process rectified this problem. Finally, glycogen accumulating organisms (GAO) were observed in BNR sludge samples, and consumption of VFA by GAO appeared to cause decreased phosphorus removal. Better phosphorus removal was obtained using VFA derived from the fermenter than dosing an equivalent amount of acetic acid. It was hypothesized that GAO have a competitive advantage to use acetate and PAO have a competitive advantage to use propionate, butyrate or some other soluble COD compound in the fermenter effluent. Contrary to popular belief, acetate may not be the optimum VFA for biological phosphorus removal. The competition between PAO and GAO for different VFA species under anaerobic conditions requires further investigation in order to control the growth of GAO and thereby improve reliability of biological phosphorus removal processes.

The effect of GAOs (glycogen accumulating organisms) on anaerobic carbon requirements in full-scale Australian EBPR (enhanced biological phosphorus removal) plants.

Glycogen-accumulating organisms (GAOs) were present in six full-scale plants investigated and in all but one made a significant contribution to the amount of volatile fatty acid (VFA) taken up anaerobically. While most plants surveyed contain GAOs, it was demonstrated that it is possible for a full-scale plant to operate with an insignificant GAO population. "Candidatus Accumulibacter phosphatis" were the significant polyphosphate-accumulating organisms (PAOs) in all plants surveyed. "Candidatus Competibacter phosphatis" were found in all plants along with other possible GAOs that were observed but not identified. A significant GAO population will increase the carbon requirements by removing VFA that could otherwise have been used by PAOs. Process optimization minimizing GAOs in full-scale plants would lead to a more efficient use of VFA.

[Proper patient counseling, recommended nutrition, specific medication. The basics of irritable bowel syndrome therapy]. / Gute Patientenführung, richtige Ernährung, gezielte Medikation. Das sind die Trümpfe in der Reizdarmtherapie.

In the treatment of the irritable bowel syndrome, it is important to qualify unrealistic expectations with regard to treatment, at an early stage. The therapeutic spectrum encompasses establishment of good rapport between physician and patient, modification of life style, provision of good patient information, reassurance, coping strategies, and temporal restraints on medication. Depending on the leading symptoms, the latter may range from laxatives to probiotics, anticholinergics or spasmolytics, prokinetic and anti-diarrheal agents, to 5-HT3/HT4 receptor antagonists. In individual patients with frequently recurrent or permanent pain, the use of tricyclic antidepressants may be considered. Painkillers should be reserved for patients in whom other therapeutic strategies have failed.

Calcium and phosphate supplementation promotes bone cell mineralization: implications for hydroxyapatite (HA)-enhanced bone formation.

Organic phosphate, in particular beta-glycerophosphate (beta-GP), has been used to induce mineralization in cell culture systems. It serves as a source of inorganic phosphate when hydrolyzed by alkaline phosphatase. This study examined the effect of supplemental calcium and phosphate as well as the influence of various metabolic inhibitors on mineralization in a rat osteoblast-like cell-culture system. Mineralization was induced by supplementation of 1.8 mM of Ca(+2) and 5 mM of beta-GP or Pi. Mineral deposits associated with in vitro mineralization were revealed under SEM and TEM. Levamisole (10-100 microM) inhibited alkaline phosphatase activity and effectively reduced mineral formation. Actinomycin (500 ng/mL) and cycloheximide (50 microg/mL) also reduced mineral depositions by blocking RNA synthesis and protein synthesis, respectively. Levamisole and beta-GP did not appear to influence DNA synthesis. Spontaneous precipitation of calcium phosphate mineral was not detected in the culture medium with calcium and phosphate supplements in the absence of cell culture. The findings suggest that an elevated concentration of calcium and phosphate is crucial for in vitro mineralization. Furthermore, the mineralization process is associated with biologic events rather than with a spontaneous precipitation of calcium phosphate mineral. In view of the degradation potential of hydroxyapatite (HA)-coated implants, these results may be a viable indication that HA enhances bone formation through a similar mechanism.

Identification of some of the major groups of bacteria in efficient and nonefficient biological phosphorus removal activated sludge systems.

To investigate the bacteria that are important to phosphorus (P) removal in activated sludge, microbial populations were analyzed during the operation of a laboratory-scale reactor with various P removal performances. The bacterial population structure, analyzed by fluorescence in situ hybridization (FISH) with oligonucleotides probes complementary to regions of the 16S and 23S rRNAs, was associated with the P removal performance of the reactor. At one stage of the reactor operation, chemical characterization revealed that extremely poor P removal was occurring. However, like in typical P-removing sludges, complete anaerobic uptake of the carbon substrate occurred. Bacteria inhibiting P removal overwhelmed the reactor, and according to FISH, bacteria of the beta subclass of the class Proteobacteria other than beta-1 or beta-2 were dominant in the sludge (58% of the population). Changes made to the operation of the reactor led to the development of a biomass population with an extremely good P removal capacity. The biochemical transformations observed in this sludge were characteristic of typical P-removing activated sludge. The microbial population analysis of the P-removing sludge indicated that bacteria of the beta-2 subclass of the class Proteobacteria and actinobacteria were dominant (55 and 35%, respectively), therefore implicating bacteria from these groups in high-performance P removal. The changes in operation that led to the improved performance of the reactor included allowing the pH to rise during the anaerobic period, which promoted anaerobic phosphate release and possibly caused selection against non-phosphate-removing bacteria.

Tumour oxygenation assessed by polarographic needle electrodes and bioenergetic status measured by 31P magnetic resonance spectroscopy in human soft tissue tumours.

The purpose of this study was to evaluate the feasibility of polarographic oxygen electrode measurements and phosphorus magnetic resonance spectroscopy (31P-MRS) in extravisceral soft tissue tumours, designated to receive preoperative radiotherapy. Pretreatment tumour oxygenation was determined in 41 cases and 31P-MRS was amenable to lesions in 34 patients. Biopsies were characterized histopathologically as 25 primary soft tissue sarcomas (STS), 2 recurrent STS, 9 benign and 5 other malignancies. Evaluation of phosphorus (31P) spectra was possible in 11 cases. The oxygenation status of normal tissue was higher than that of tumours, whereas no difference was found between oxygenation status of benign lesions and that of STS. There was substantial variation between tumours in the median pO2 and the bioenergetic status (beta-NTP/Pi). No correlation was found between tumour pO2 and volume (n = 25). Moreover, there was no correlation between beta-NTP/Pi and the median tumour pO2, the fraction of pO2 values < or =2.5 mmHg or tumour volume (n = 10), respectively. In conclusion, oxygen electrode assessment was found to be a clinically applicable and feasible technique for measuring tumour oxygenation status, whereas the success of 31P-MRS in human neoplasms was limited by a very poor resolution in the phosphorus signal that allowed analysis of 31P spectra in 11 tumours out of 34 cases.