Upgrading of sewage treatment plant by sustainable and cost-effective separate treatment of industrial wastewater.

The Olburgen sewage treatment plant has been upgraded to improve the effluent quality by implementing a separate and dedicated treatment for industrial (potato) wastewater and reject water. The separate industrial treatment has been realized within a beneficial public-private partnership. The separate treatment of the concentrated flows of industrial wastewater and sludge treatment effluent proved to be more cost-efficient and area and energy efficient than a combined traditional treatment process. The industrial wastewater was first treated in a UASB reactor for biogas production. The UASB reactor effluent was combined with the reject water and treated in a struvite reactor (Phospaq process) followed by a one stage granular sludge nitritation/anammox process. For the first time both reactors where demonstrated on full scale and have been operated stable over a period of 3 years. The recovered struvite has been tested as a suitable substitute for commercial fertilizers. Prolonged exposure of granular anammox biomass to nitrite levels up to 30 mg/l did not result in inhibition of the anammox bacteria in this reactor configuration. The chosen option required a 17 times smaller reactorvolume (20,000 m(3) less volume) and saves electric power by approximately 1.5 GWh per year.

Peptide-mediated gene transfer of cationic lipid/plasmid DNA complexes to endothelial cells.

The purpose of this research is to develop ligand-targeted plasmid based gene delivery systems for gene transfer to tumor endothelium. Cell adhesion assays were used to test the peptide inhibition of human endothelial cell adsorption to vitronectin-treated tissue culture plates. A series of RGD containing peptides were tested in linear form and with one and two disulfide bonds. The linear and two disulfide bond peptides yielded similar IC50 (approximately 1 x 10(-7) M). Substitution of two methionines for cysteines yielded a single disulfide bond that increased the IC50 by 10-fold. The single and double disulfide peptides were derivatized to N-succinyl-dioleoylphopsphatidylethanolamine and incorporated into 100 nm liposomes radiolabeled with H-cholesterylhexadecylether. Liposome uptake by human umbilical vein endothelial cells was tested as a function of lipopeptide surface density. Increase in membrane surface density from 5 to 20mol% increased human umbilical derived endothelial cell (HUVEC) uptake of the liposomes for both the single and double disulfide peptides. Liposome uptake by HUVECs was 3-fold greater for the double disulfide compared to the single disulfide. The single and double disulfide lipopeptides were then tested for gene transfer to HUVECs using DOTMA:Cholesterol cationic liposomes. The polyplexes were formed by rapidly mixing plasmid DNA with DOTMA:CHOL liposomes at a 3:1 charge ratio in 2% ethanol, 10% lactose. The ethanol was removed by lyophilization and upon rehydration, the lipoplexes had a mean diameter of approximately 100nm. HUVEC transfection studies showed that increasing the mol% of the single disulfide RGD lipopeptide to 20mol% increased gene transfer by 10-fold. This increase in transfection could be reduced to that obtained in the absence of lipopeptide by co-incubating the HUVECs with a 100-fold excess of the single disulfide RGD peptide, thus demonstrating lipopeptide mediated gene transfer to endothelial cells.

DC-SIGN-specific liposomal targeting and selective intracellular compound delivery to human myeloid dendritic cells: implications for HIV disease.

Myeloid dendritic cells (MyDCs), prime stimulators of antigen-specific immunity, can serve as one of the major reservoirs for human immunodeficiency virus type-1 (HIV-1). Utilizing mature monocyte-derived MyDCs generated with granulocyte/macrophage colony-stimulating factor, interleukin-4, and tumour necrosis factor-alpha as an in vitro model, we here present the first proof of concept for liposomal compound delivery to these cells by specifically addressing CD209, i.e. DC-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN), a MyDC-associated C-type lectin implicated in the transmission of HIV-1 to T helper cells. By employing a liposomally entrapped tracer, calcein, we demonstrate by flow cytometry and mathematics a superior targeting efficacy for DC-SIGN, as compared with select other MyDC markers (CD1a, CD4, CD45R0, and CD83). Fluorescence microscopy reveals time-dependent surface binding and intracellular uptake of DC-SIGN-specific liposomes by both immature and mature MyDCs. This pilot study implies that liposomal targeting to CD209 and related C-type lectins may afford therapeutic intracellular drug delivery to MyDCs and other reservoir and nonreservoir cells susceptible to infection with HIV-1.

Tuning the recovery of an Rh-containing catalyst with silica-based (poly) amine ion exchangers.

Highly efficient Rh-recovery from different adsorption media has been effected with silica-based (chelating) ion exchangers containing (poly) amine functionalities; recoveries have been found to correlate well with the stability of the metal-to-ligand complexes.

Spontaneous regression of hepatocellular carcinoma.

A 72-year-old Caucasian woman suffered from histologically-proven advanced hepatic cancer, for which she received no treatment. She had been a regular drinker for a long time. Serologic markers for hepatitis B and C were negative. In spite of her poor prognosis, she remained in good clinical condition and at 14 months of follow up the hepatocellular carcinoma could not be visualised any more radiologically. At that time the serum alpha foetoprotein concentration was normal. At present, 28 months after diagnosis, the patient is doing well and her tumour still appears to be in complete spontaneous remission.

Effectiveness of quadruple therapy using lansoprazole, instead of omeprazole, in curing Helicobacter pylori infection.

BACKGROUND: Omeprazole enhances the efficacy of bismuth-based triple therapy. It is unknown whether the same is true for other proton pump inhibitors. Lansoprazole has superior anti-Helicobacter activity in vitro and possibly also in vivo; therefore we investigated quadruple therapy with lansoprazole. MATERIALS AND METHODS: In two studies performed in separate hospitals, a total of 67 Helicobacter pylori-positive patients were treated with 7-day quadruple therapy (lansoprazole, colloidal bismuth subcitrate, tetracycline, and metronidazole) after 3 days of lansoprazole pretreatment. Testing for cure was done by endoscopy in study 1 and by breath test in study 2. RESULTS: Cure rates per protocol were 31 of 31 (100%) in study 1 and 30 of 32 (94%) in study 2. Intention-to-treat cure rates were 31 of 35 (89%) in study 1 and 30 of 32 (94%) in study 2. Cured overall were 32 of 34 with a metronidazole sensitive strain and 3 of 3 with a metronidazole-resistant strain. Data on side effects were collected from 51 patients. Twelve (21%) had no side effects, 27 (53%) had mild side effects, 10 (20%) had moderate side effects, but only 2 (4%) had severe side effects. Side effects, never were the reason that a patient stopped taking the medication. CONCLUSIONS: The results with lansoprazole-quadruple therapy are comparable to the historic control group treated with omeprazole-quadruple therapy. The cure rare is very high, and although mild to moderate side effects occurred in many patients, everybody finished the treatment regime.

Long-term follow-up after cure of Helicobacter pylori infection with 4 days of quadruple therapy.

BACKGROUND: We have shown that 4 days of quadruple therapy after omeprazole pre-treatment is an effective therapy for curing H. pylori infection. In this study we investigated whether this regimen would maintain the high cure rate during long-term follow-up. Some recent studies have reported high recurrence rates after apparent cure. Apparently not all methods to test for cure have sufficient sensitivity to pick up small numbers of residual bacteria. This study also served to investigate whether our methods to test for cure 5-6 weeks post-treatment were reliable. METHODS: All patients from a previous study were invited to return for a 14C-urea breath test and serology. A representative group of 37 patients (76%) returned for a urea breath test and serology. The mean follow-up was 14.7 months (range 11.4-23.6 months). RESULTS: None of the 37 patients had a positive urea breath test results. IgG antibody titres fell steadily in all patients, showing a mean decrease of 83% at the end of the follow-up. None of the patients showed an increase in titre. Reinfection was therefore 0% (0 of 37). CONCLUSION: Four days of quadruple therapy seems to be an effective therapy for the eradication of H. pylori as evidenced after long-term follow-up. Our biopsy methodology is reliable in identifying treatment failures 5-6 weeks post-treatment.

Regional enteritis complicating malignant lymphoma.

We describe a patient presenting with palpable lymph nodes due to non-Hodgkin's lymphoma. Chemotherapy induced complete remission. One year later he complained of cramping abdominal pain, diarrhoea and bloody stools all due to Crohn's disease of the colon. There are only a few more patients described with a combination of inflammatory bowel disease and malignant lymphoma. So far there is no explanation for the co-incidence of the two diseases.

Quadruple therapy compared with dual therapy for eradication of Helicobacter pylori in ulcer patients: results of a randomized prospective single-centre study.

OBJECTIVE: To assess the efficacy and side-effect profile of two currently advocated treatment regimens for eradicating Helicobacter pylori. DESIGN: A randomized, controlled, open, single-centre study. SETTING: A community hospital in The Netherlands. PARTICIPANTS: Seventy-six consecutive patients with (chronic) ulcer disease and biopsy-proven H. pylori infection, but without active ulceration at the time of inclusion. INTERVENTIONS: Patients were randomly allocated to 1 week of quadruple therapy with omeprazole, bismuth, tetracycline and metronidazole (group 1) or 2 weeks of dual therapy with omeprazole and amoxicillin (group 2). Group 1 patients were pretreated with omeprazole for 3 days. MAIN OUTCOME MEASURES: Cure was confirmed by obtaining 10 endoscopic biopsies for urease testing, histology and culture 6 weeks after treatment. Side-effects were scored on a standard questionnaire. RESULTS: Three patients were lost to follow-up. In the 'intention to treat' analysis 37 (92.5%) of 40 patients in group 1 were cured compared with 20 (55.6%) of 36 patients in group 2 (P < 0.001). The difference in efficacy was 36.9% (95% confidence interval 18.7-55.1%). Side-effects were fewer and milder in group 2, but all patients in both groups were able to complete the course of treatment. CONCLUSION: Dual therapy is significantly less effective in curing H. pylori infection in peptic ulcer patients than quadruple therapy. No patients were intolerant to either treatment. On the basis of the low efficacy of dual therapy, we believe that this therapy should not be used as a first-line treatment strategy. We confirmed our previous finding that 1 week of quadruple therapy is tolerated well and that it is highly effective against metronidazole-sensitive as well as metronidazole-resistant strains of H. pylori.

Only four days of quadruple therapy can effectively cure Helicobacter pylori infection.

AIM: To determine whether 4 days of quadruple therapy using bismuth, tetracycline and metronidazole combined with omeprazole is effective treatment for Helicobacter pylori infection. METHODS: Non-ulcer dyspepsia, as well as chronic peptic ulcer patients with biopsy-proven H. pylori infection received 4 days of quadruple therapy. They were pretreated with 3 days of omeprazole. At least 5-6 weeks later, endoscopy was repeated with 10 biopsies for urease test, histology and culture to establish cure of infection. RESULTS: None of the 54 patients included was lost to follow-up but two had a 14C-urea breath test instead of endoscopy. Side-effects did not interfere with compliance. Forty-nine out of 54 patients (91%; 95% CI: 80-97%) were cured. Metronidazole susceptibility data were available from 43 pre-treatment isolates. Of these 38/40 (95%) with a metronidazole-sensitive strain, and one of three with a metronidazole-resistant strain were cured. CONCLUSIONS: Four days of quadruple therapy after omeprazole pre-treatment is a feasible, well tolerated, and effective treatment for H. pylori infection, especially in those carrying a metronidazole-sensitive strain. It seems that in quadruple therapy, cure rate and treatment duration have a non-linear relation. Our results need confirmation, but for patients suffering from side-effects with the 7-day regimen stopping treatment after 4 days is justified.

Effect of acid suppression on efficacy of treatment for Helicobacter pylori infection.

Eradication of Helicobacter pylori from the stomach by triple therapy with bismuth, tetracycline, and metronidazole cures peptic ulcer disease. We investigated whether concomitant acid inhibition with omeprazole would improve the results of triple therapy. 108 consecutive patients with peptic-ulcer disease and biopsy-proven H pylori infection were randomised to 7 days of triple therapy with or without omeprazole 20 mg twice daily. Patients in the omeprazole-treated group were pretreated with 3 days of omeprazole. Eradication of H pylori was assessed by 10 endoscopic biopsies for urease test, histology, and culture 4-6 weeks after treatment. 53 of 54 (98.1%) patients treated with omeprazole were cured compared with 45 of 54 (83.3%) of those not treated (p = 0.02), a difference in efficacy of 14.8% (95% Cl 4.2-25.4%). Most side effects were mild and did not interfere with compliance; 105 patients (97.2%) finished treatment. Gastro-intestinal side effects were significantly fewer in the omeprazole group. We conclude that the addition of omeprazole to triple therapy improves efficacy, lessens side effects, and is sufficiently efficacious to obviate the need for a diagnostic test of cure in compliant patients.

[Methotrexate as possible treatment in refractory chronic inflammatory intestinal disease]. / Methotrexaat als mogelijke behandeling bij refractaire chronische inflammatoire darmziekte.

OBJECTIVE: To determine the value of low dose methotrexate therapy in chronic inflammatory bowel disease. SETTING: Catharina Hospital and Diaconessenhuis, Eindhoven, and St. Joseph Hospital, Veldhoven. DESIGN: Descriptive. METHOD: From 1988 until 1993 we treated 15 patients 16 times (one patient was treated twice) with methotrexate 25 mg i.m. once a week during 12 weeks, followed by a tapering oral dose. Our population consisted of 4 men and 11 women with a mean age of 31 year. The diagnosis was Crohn ileitis (1 patient), Crohn colitis (8), ileocolitis (4) and ulcerative colitis (2). The indication for methotrexate was resistance to therapy (9) and steroid dependency (7). Retrospectively the disease activity was determined after 1, 2 and 3 months of therapy. RESULTS: The mean defaecation frequency went down from 7 to 2 times daily after 12 weeks, the ESR from 47 to 17 mm/1st hour, the thrombocytes from 436 x 10(9)/l to 325 x 10(9)/l and the prednisone dose could be lowered from 22 mg to 15 mg after 3 months. In 13/16 treatment cases there was a subjective positive response to methotrexate. Initial response was seen within 4 weeks. No serious side effects were seen. After 10 of the 13 response episodes the disease recurred. CONCLUSION: Methotrexate 25 mg once a week i.m. was associated with a subjective and objective improvement in 12/15 patients, but the risk of recurrence after tapering appears to be large.

Randomized study comparing 1 with 2 weeks of quadruple therapy for eradicating Helicobacter pylori.

OBJECTIVES: We investigated whether the recommended 2-wk triple therapy for eradicating Helicobacter pylori could be reduced to 1 wk, and thus we tried to determine the optimal treatment duration for triple therapy. METHODS: A group of 111 consecutive patients with H. pylori-proven chronic peptic ulcer disease was entered in a randomized study comparing the side effects, compliance, and eradication rates with either 1 wk or 2 wk of traditional triple therapy in combination with an H2 receptor antagonist (quadruple therapy). RESULTS: Follow-up data were available for 109 patients. Eradication for 1 wk of treatment was 53/56 (95%) [95% confidence interval (CI), 89-100%] and for the 2-wk treatment schedule was 50/53 (94%) (95% CI, 88-100%). Therefore 1-wk quadruple therapy appears sufficient, and prolonging treatment does not increase efficacy. Compliance with quadruple therapy is better when the duration of treatment decreases from 2 to 1 wk. One-week quadruple therapy seems feasible for most patients. CONCLUSIONS: Because of the high eradication rate and good tolerability of this short course of quadruple therapy, we recommend it as first-line anti-Helicobacter treatment in a compliant population.

Role of the intestinal microflora in the formation of DNA and haemoglobin adducts in rats treated with 2-nitrofluorene and 2-aminofluorene by gavage.

The role of the intestinal microflora in the metabolic activation of nitroarenes and arylamines was studied in female Wistar rats that received a dose of 1 mmol/kg 2-aminofluorene (2-AF) in sunflower oil by gavage. Another group received the same dose of 2-nitrofluorene (2-NF). A third group of animals was used as controls. Germfree (GF) rats, GF rats with a rat microflora (RM) and GF rats with a human microflora (HM) were treated. After treatment with 2-AF significant differences were observed in the formation of haemoglobin (Hb) adducts and DNA adducts. The 2-AF-Hb adduct level (mean +/- SD) observed in GF rats (0.57 +/- 0.13 mumol/g Hb) was considerably lower than that observed in RM rats (5.1 +/- 0.6) and in HM rats (6.2 +/- 1.3). DNA adduct levels showed the opposite pattern: levels of adducts co-migrating with deoxyguanosin-8-yl-aminofluorene (dG-C8-AF) in liver tissue were higher in GF rats (4.6 +/- 1.4 fmol/micrograms DNA) as compared to RM rats (2.6 +/- 0.04) or HM rats (2.0 +/- 0.7). In lung tissue and white blood cells a similar influence of the intestinal microflora on DNA adduct levels was observed. These results suggest that the intestinal microflora cleaves conjugates of 2-AF or N-hydroxy-2-AF, thus facilitating enterohepatic recirculation of these compounds and enhancing the formation of reactive intermediates binding to Hb. The latter is not observed for DNA adduct formation, indicating that most of these adducts have been formed after a single passage through the liver. After treatment with 2-NF, Hb and DNA adduct levels were much lower. An adduct spot was observed that was not present in rats that received 2-AF. In GF animals only very low levels of DNA adducts co-migrating with dG-C8-AF or deoxyguanosin-8-yl-acetyl-aminofluorene and no Hb adducts were observed, indicating that the metabolic activity of the microflora is an essential step in both Hb and DNA adduct formation.

[Endoscopic studies of the digestive tract as a service for family practitioners; experience in the Eindhoven area]. / Endoscopisch onderzoek van de tractus digestivus als service voor de huisarts; ervaringen in de regio Eindhoven.

In May 1989 an open access endoscopy service for general practitioners (GPs) was started in the region of Eindhoven. In the four hospitals covering a population of 500,000 inhabitants, served by 200 GPs, uniform working agreements were made. We evaluated the endoscopies performed in 1990 and compared the total number of endoscopies with that of gastrointestinal X-rays. A total of 996 open access gastroscopies were performed. The most important diagnoses were: peptic ulcer (19%), oesophagitis (16%), bulbitis (9%) and carcinoma (1.5%). The diagnostic yield was 57%. A total of 509 GP-referred sigmoidoscopies were performed. The most important diagnoses were: haemorrhoids and anal fissure (28%), colitis (15%), diverticulosis (12%), neoplastic polyps (11%) and carcinoma (4%). The diagnostic yield was 77%. Compared with 1989 the total number of gastroscopies, requested by GPs and by specialists, increased by 15% of which two-thirds were requested by the GPs. In this period the reduction of barium meals was 11% which was almost exclusively due to the specialist group. Sigmoidoscopies increased by 7%, entirely due to the GP group. The number of barium enemas remained constant. Open access endoscopy resulted in an increase endoscopies with only a partial substitution of radiological examinations. The increase in endoscopic examinations in the GP group did not result in a decline in the specialist group. The total diagnostic yield was 64%.

Two sisters with coeliac disease and jejunal cancer: just a coincidence?

Two sisters with asymptomatic coeliac disease are described; they both developed a primary jejunal cancer at the same age. While screening the family, a third sister was found to have coeliac disease, but without detectable cancer in the small intestine or stomach. These findings suggest an increased susceptibility in this family for carcinoma developing secondary to asymptomatic coeliac disease. We conclude that the finding in a patient of the combination of coeliac disease and malignancy in the digestive tract is sufficient reason to investigate the first-degree relatives with regard to the presence of coeliac disease and a secondary carcinoma.