Elastic turbulence in entangled semi-dilute DNA solutions measured with optical coherence tomography velocimetry.

The flow instabilities of solutions of high molecular weight DNA in the entangled semi-dilute concentration regime were investigated using optical coherence tomography velocimetry, a technique that provides high spatial (probe volumes of 3.4 pL) and temporal resolution (sub µs) information on the flow behaviour of complex fluids in a rheometer. The velocity profiles of the opaque DNA solutions (high and low salt) were measured as a function of the distance across the gap of a parallel plate rheometer, and their evolution over time was measured. At lower DNA concentrations and low shear rates, the velocity fluctuations were well described by Gaussian functions and the velocity gradient was uniform across the rheometer gap, which is expected for Newtonian flows. As the DNA concentration and shear rate were increased there was a stable wall slip regime followed by an evolving wall slip regime, which is finally followed by the onset of elastic turbulence. Strain localization (shear banding) is observed on the boundaries of the flows at intermediate shear rates, but decreases in the high shear elastic turbulence regime, where bulk strain localization occurs. A dynamic phase diagram for non-linear flow was created to describe the different behaviours.

Nasopharyngeal vs. adenoid cultures in children undergoing adenoidectomy: prevalence of bacterial pathogens, their interactions and risk factors.

Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus colonization of the adenoids and nasopharynx in 103 preschool children who underwent adenoidectomy for recurrent upper respiratory tract infections was examined. Bacterial interactions and risk factors for bacterial colonization of the nasopharynx and adenoids, separately, were analysed statistically. The prevalence of simultaneous isolation from both anatomical sites was 45·6% for S. pneumoniae, 29·1% for H. influenzae, 15·5% for M. catarrhalis and 18·4% for S. aureus. Three pathogens were significantly more frequent together from adenoid samples; nasopharyngeal swabs more often yielded a single organism, but without statistical significance. M. catarrhalis and S. aureus significantly more frequently co-existed with S. pneumoniae and H. influenzae than with each other and a positive association of S. pneumoniae and H. influenzae in adenoid samples was evident. Several differences between risk factors for nasopharyngeal and adenoid colonization by the individual pathogens were observed. We conclude that the adenoids and nasopharynx appear to differ substantially in colonization by pathogenic microbes but occurrence of H. influenzae and S. pneumoniae in the nasopharynx could be predictive of upper respiratory tract infections.

Optical coherence tomography velocimetry of colloidal suspensions.

Optical coherence tomography velocimetry combined with a rheometer and optical modulation techniques provides increased sensitivity to the low shear rate motion of complex fluid systems. Optical modulation coupled with a new interferometer design yields improved signal to noise ratios and is demonstrated with optically opaque colloidal suspensions. Thus the measurable range of shear velocities with complex fluids can be as low as ∼40 µm s(-1), more than an order of magnitude improvement on the previous lower limit of ∼700 µm s(-1). Furthermore the apparatus demonstrates improved sensitivity to the measurement of velocity. The instrument was used to study two hard sphere colloidal systems, sterically stabilized PVP spheres of 1 µm radius and sterically stabilized polystyrene spheres of 600 nm radius, which display shear banding behavior due to shear induced concentration gradients. OCT velocimetry also allows the velocity fluctuations of the system to be quantified as a function of the distance across the rheometer gap to help classify underlying unsteady or turbulent phenomena.

Exposure to candesartan during the first trimester of pregnancy in type 1 diabetes: experience from the placebo-controlled DIabetic REtinopathy Candesartan Trials.

AIMS/HYPOTHESIS: The teratogenic consequences of angiotensin-converting enzyme inhibitors angiotensin receptor blockers (ARBs) during the second and third trimesters of pregnancy are well described. However, the consequences of exposure during the first trimester are unclear, especially in diabetes. We report the experience from DIRECT (DIabetic REtinopathy and Candesartan Trials), three placebo-controlled studies designed to examine the effects of an ARB, candesartan, on diabetic retinopathy. METHODS: Over 4 years or longer, 178 normotensive women with type 1 diabetes (86 randomised to candesartan, 32 mg once daily, and 92 assigned to placebo) became pregnant (total of 208 pregnancies). RESULTS: More than half of patients were exposed to candesartan or placebo prior to or in early pregnancy, but all discontinued it at an estimated 8 weeks from the last menstrual period. Full-term pregnancies (51 vs 50), premature deliveries (21 vs 27), spontaneous miscarriages (12 vs 15), elective terminations (15 vs 14) and other outcomes (1 vs 2) were similar in the candesartan and placebo groups. There were two stillbirths and two 'sick babies' in the candesartan group, and one stillbirth, eight 'sick babies' and one cardiac malformation in the placebo group. CONCLUSIONS/INTERPRETATION: The risk for fetal consequences of ARBs in type 1 diabetes may not be high if exposure is clearly limited to the first trimester. Long-term studies in fertile women can be conducted with ARBs during pregnancy, provided investigators diligently stop their administration upon planning or detection of pregnancy. TRIAL REGISTRATION: ClinicalTrials.gov DIRECT-Prevent 1 NCT00252733; DIRECT-Protect 1 NCT00252720; DIRECT-Protect 2 NCT00252694. FUNDING: The study was funded jointly by AstraZeneca and Takeda.

Seven-year follow-up of a randomized clinical trial comparing proton-pump inhibition with surgical therapy for reflux oesophagitis.

BACKGROUND: This randomized clinical trial compared long-term outcome after antireflux surgery with acid inhibition therapy in the treatment of chronic gastro-oesophageal reflux disease (GORD). METHODS: Patients with chronic GORD and oesophagitis verified at endoscopy were allocated to treatment with omeprazole (154 patients) or antireflux surgery (144). After 7 years of follow-up, 119 patients in the omeprazole arm and 99 who had antireflux surgery were available for evaluation. The primary outcome variable was the cumulative proportion of patients in whom treatment failed. Secondary objectives were evaluation of the treatment failure rate after dose adjustment of omeprazole, safety, and the frequency and severity of post-fundoplication complaints. RESULTS: The proportion of patients in whom treatment did not fail during the 7 years was significantly higher in the surgical than in the medical group (66.7 versus 46.7 per cent respectively; P=0.002). A smaller difference remained after dose adjustment in the omeprazole group (P=0.045). More patients in the surgical group complained of symptoms such as dysphagia, inability to belch or vomit, and rectal flatulence. These complaints were fairly stable throughout the study interval. The mean daily dose of omeprazole was 22.8, 24.1, 24.3 and 24.3 mg at 1, 3, 5 and 7 years respectively. CONCLUSION: Chronic GORD can be treated effectively by either antireflux surgery or omeprazole therapy. After 7 years, surgery was more effective in controlling overall disease symptoms, but specific post-fundoplication complaints remained a problem. There appeared to be no dose escalation of omeprazole with time.

Changes of gastric mucosal architecture during long-term omeprazole therapy: results of a randomized clinical trial.

BACKGROUND: The impact of long-term acid suppression on the gastric mucosa remains controversial. AIM: To report further observations on an established cohort of patients with gastro-oesophageal reflux disease, after 7 years of follow-up. METHODS: Of the original cohort randomized to either antireflux surgery or omeprazole, 117 and 98 patients remained in the medical and surgical arms, respectively. Gastric biopsies were taken at baseline and throughout the study. RESULTS: Fifty-three antireflux surgery and 39 omeprazole-treated patients had Helicobacter pylori infection at randomization. Eighty-three omeprazole-treated and 60 antireflux surgery patients remained H. pylori negative over the 7 years, and no change was observed in mucosal morphology except for a change in endocrine cell population (linear and diffuse hyperplasia, P = 0.03). During the 7-year study many patients, who were initially H. pylori infected, had the infection eradicated leaving only 13 omeprazole and 12 antireflux surgery patients still infected. In these patients, omeprazole induced a deterioration of the mucosal inflammation scores (P = 0.01) with a numerical increase of glandular atrophy. CONCLUSIONS: Long-term omeprazole therapy does not alter the exocrine oxyntic mucosal morphology in H. pylori-negative patients, but mucosal endocrine cells appear to be under proliferative stimulation; in H. pylori-positive patients there are changes in mucosal inflammation and atrophy.

Modelling non-structural best management practices--focus on reductions in stormwater pollution.

This paper describes a modelling approach for evaluating the efficiency of different non-structural best management practices for stormwater management. A scenario with a set of source reduction practices was simulated using the substance flow model SEWSYS for an urban catchment in the city of Göteborg, Sweden. The scenario is based on a hypothetical control program that includes prevention, education and regulations. The simulation shows relatively high reductions of copper and PAH, 77% and 50%, respectively. The reduction in copper is mainly due to less copper roof corrosion and brake wear, while reduced road wear has the greatest effect for PAH. An important result from this study is that the nonstructural BMPs applied did not give a sufficient reduction in pollution to meet the desirable environmental quality criteria. To meet these criteria, additional BMPs must be implemented, preferably a combination of both non-structural and structural measures.

Effects of pregnancy on mothers' sleep.

OBJECTIVES: To survey the effects of pregnancy on mothers' sleep. METHODS: Mothers were interviewed during and after pregnancy with a series of five questionnaires to assess alterations in their sleep. The first questionnaire covered the 3 months before becoming pregnant, the next three the trimesters of pregnancy and the last the 3 months after delivery. The study was carried out in a central hospital and the maternity care units in the nearby rural community. Altogether, 325 pregnant women completed all five questionnaires. RESULTS: The total amounts of reported sleep and of nocturnal sleep increased significantly during the first trimester of pregnancy, began to decrease thereafter and were shortest during the 3 months after pregnancy. During late pregnancy expectant mothers over 30 years of age reported less sleep than those under 30. During the whole pregnancy, but increasingly toward the end of pregnancy, sleep became more restless and fragmentary and its subjective quality worsened, due at least partly to increased restless legs and nightly awakenings increasing with advancing pregnancy. CONCLUSIONS: The subjective quality of sleep is disturbed as early as the first trimester of pregnancy, although total sleeping time increases. The amount of reported sleep begins to decrease in the second trimester. The frequency of reported sleep disturbances, such as restless legs syndrome and nocturnal awakenings, is maximum in the third trimester but is about normal within 3 months after delivery.

Energy conservation in aerobically grown Staphylococcus aureus.

The present studies provide new data on the involvement of menaquinol oxidases in substrate oxidation and energy conservation in aerobically grown, resting cells of Staphylococcus aureus 17810R, starved of endogenous energy reserves and supplemented with glutamate or L-lactate. These cells were energetically competent, since they oxidized both substrates, generated an electrochemical proton gradient (deltamuH+) and synthesized ATP via oxidative phosphorylation. Studies with KCN showed that: (i) L-lactate oxidation occurred via two terminal menaquinol oxidases - the ba3-type sensitive to low KCN and the bo-type insensitive to cyanide, (ii) glutamate oxidation proceeded via the bo-type oxidase, and (iii) ATP synthesis with glutamate or L-lactate was coupled only to the bo-type oxidase. Also in glucose-grown cells oxidizing L-lactate, ATP synthesis was coupled to the highly repressed bo-type oxidase. It is suggested that in the respiratory chain of strain 17810R two energy coupling sites may be present: in the complex of NADH-menaquinone oxidoreductase and in the complex of the bo-type menaquinol oxidase. The rate of ATP synthesis was similar with both substrates, but the rate of their oxidation differed significantly: the P/O ratios were 1.5 and 0.03 with glutamate and L-lactate, respectively. CCCP accelerated glutamate oxidation by 50% but was without effect on L-lactate oxidation. In cell lysates, the rates of NADH and L-lactate oxidation were equal. It is concluded that in whole cells of S. aureus 17810R oxidation of NADH derived from glutamate breakdown is tightly coupled to phosphorylation, while L-lactate oxidation seems to be rather loosely coupled.

Tumor necrosis factor-alpha, sphingomyelinase, and ceramide inhibit store-operated calcium entry in thyroid FRTL-5 cells.

Tumor necrosis factor alpha (TNF-alpha) is a potent inhibitor of proliferation in several cell types, including thyroid FRTL-5 cells. As intracellular free calcium ([Ca2+]i) is a major signal in activating proliferation, we investigated the effect of TNF-alpha on calcium fluxes in FRTL-5 cells. TNF-alpha per se did not modulate resting [Ca2+]i. However, preincubation (10 min) of the cells with 1-100 ng/ml TNF-alpha decreased the thapsigargin (Tg)-evoked store-operated calcium entry in a concentration-dependent manner. TNF-alpha did not inhibit the mobilization of sequestered calcium. To investigate whether the effect of TNF-alpha on calcium entry was mediated via the sphingomyelinase pathway, the cells were pretreated with sphingomyelinase (SMase) prior to stimulation with Tg. SMase inhibited the Tg-evoked calcium entry in a concentration-dependent manner. Furthermore, an inhibition of calcium entry was obtained after preincubation of the cells with the membrane-permeable C2-ceramide and C6-ceramide analogues. The inactive ceramides dihydro-C2 and dihydro-C6 showed only marginal effects. Neither SMase, C2-ceramide, nor C6-ceramide affected the release of sequestered calcium. C2- and C6-ceramide also decreased the ATP-evoked calcium entry, without affecting the release of sequestered calcium. The effect of TNF-alpha and SMase was inhibited by the kinase inhibitor staurosporin and by the protein kinase C (PKC) inhibitor calphostin C but not by down-regulation of PKC. However, we were unable to measure a significant activation of PKC using TNF-alpha or C6-ceramide. The effect of TNF-alpha was not mediated via activation of either c-Jun N-terminal kinase or p38 kinase. We were unable to detect an increase in the ceramide (or sphingosine) content of the cells after stimulation with TNF-alpha for up to 30 min. Thus, one mechanism of action of TNF-alpha, SMase, and ceramide on thyroid FRTL-5 cells is to inhibit calcium entry.

Substrate-dependent cadmium toxicity affecting energy-linked K+/86Rb transport in Staphylococcus aureus.

Bacteria accumulate high amounts of potassium in the cytoplasm. For studying transport of K+ (with 86Rb as a marker) in bacteria (Staphylococcus aureus 17810S), the cells were depleted of the internal K+ pool by a DNP treatment. Kinetics and energetics of 86Rb transport was assayed with glucose as an exogenous energy source. It was shown that 86Rb uptake proceeded via a low affinity K+ transport system with an apparent K(m) of 2.3 mmol/L Rb+. Studies with the lipophilic cation TPP+ (tetraphenylphosphonium), the protonophore CCCP (carbonyl cyanide 3-chlorophenylhydrazone) and inhibitors (HQNO--2-heptyl-4-hydroxyquinoline N-oxide; iodoacetate) indicated that 86Rb transport was driven by delta psi (membrane potential) generated via the respiratory chain. The effect of Cd2+ on 86Rb transport was assayed with two energy donors--glucose and L-lactate. It was found that Cd2+ strongly inhibited delta psi-dependent 86Rb transport energized by cadmium-sensitive glucose oxidation, but was not toxic when cadmium-insensitive L-lactate was used as an energy source. The mechanism of these differential, substrate-dependent effects of Cd2+ on 86Rb transport is discussed.

Outcome for patients with leukemia, multiple myeloma and lymphoma who relapse after high dose therapy and autologous stem cell support.

Relapses after autologous transplantation are a serious clinical problem in patients with haematological diseases. The decision making for handling of such patients is difficult and the aim of this retrospective analysis of posttransplant relapses was 1) to obtain information of practical importance for the management of future relapses and 2) to evaluate the basis for clinical phase I-II trials of salvage therapy combined with biological modifiers. Included in the study were 283 patients with acute leukemia, multiple myeloma and malignant lymphoma who relapsed after autologous transplantations during a five year period from 1989 to 1994. Chemo- and radiotherapy was given to 229 patients after relapse or due to progressive disease and the response evaluated after 90 days. Fifty four patients (24%) obtained a complete remission and 44 patients (19%) partial responses. The overall median survival from relapse was 5 months. In the group given salvage treatment the median survival was 7 months and in the 54 patients who obtained remission the median survival was 15 months. So far 6 of 14 patients in continuous complete remission have a remission time after relapse longer than the time in remission after transplantation. Survival after relapse depended upon the time from transplantation to relapse, primary disease and if salvage therapy was given. In conclusion posttransplant relapses can be treated but the strategy has to be evaluated in future clinical trials.

Cadmium-sensitive targets in the aerobic respiratory metabolism of Staphylococcus aureus.

Studies on the effect of various Cd2+ concentrations on substrate oxidation by whole cells of cadmium-sensitive Staphylococcus aureus 17810S showed that oxidation of glutamate or pyruvate was highly sensitive to low Cd2+ concentrations (5 microM), whereas L-lactate oxidation was insensitive even to high Cd2+ concentrations (100 microM). Location of the cadmium-sensitive targets in the enzyme systems involved in oxidation of these substrates was studied in subcellular fractions prepared from cells pretreated with 5 or 100 microM Cd2+. Activities of the cytoplasmic 2-oxoglutarate dehydrogenase complex (ODHC)') and pyruvate dehydrogenase complex (PDHC) were strongly inhibited with 5 microM Cd2+, while with 100 microM Cd2+ the inhibition was almost complete. In contrast, activities of the cytoplasmic NAD-dependent glutamate dehydrogenase (NAD-GDH), the membrane-bound NADH dehydrogenase (NDH) and HQNO-sensitive NADH oxidase were not sensitive to 100 microM Cd2+. These data indicate that the accessible, cadmium-sensitive targets are located only in the cytoplasmic ODHC and PDHC. It is postulated that two vicinal dithiols present in ODHC and PDHC may be regarded as the primary cadmium-sensitive targets in the systems oxidizing glutamate or pyruvate. Since activities of the membrane-bound NAD-independent L-lactate dehydrogenase (iLDH) and HQNO-sensitive L-lactate oxidase were not affected by 100 microM Cd2+, this indicates that the L-lactate oxidizing system lacks the accessible, cadmium-sensitive targets. The mechanism of Cd2+ toxicity to energy conservation with glutamate, pyruvate or L-lactate in S. aureus is discussed.

Energetic basis of cadmium toxicity in Staphylococcus aureus.

In washed cells of cadmium-sensitive Staphylococcus aureus 17810S oxidizing glutamate, initial Cd2+ influx via the Mn2+ porter down membrane potential (delta psi) was fast due to involvement of energy generated by two proton pumps--the respiratory chain and the ATP synthetase complex working in the hydrolytic direction. Such an unusual energy drain for rapid initial Cd2+ influx is suggested to be due to a series of toxic events elicited by Cd2+ accumulation down delta psi generated via the redox proton pump: (i) strong inhibition of glutamate oxidation accompanied by a decrease of electrochemical proton gradient (delta mu H+) formation via the respiratory chain, (ii) automatic reversal of ATP synthetase from biosynthetic to hydrolytic mode, which was monitored by a decrease of delta mu (H+)-dependent ATP synthesis, (iii) acceleration of the initial Cd2+ influx down delta psi generated by the reversed ATP synthetase, the alternative proton pump hydrolyzing endogenous ATP. The primary, cadmium-sensitive targets in strain 17810S seem to be dithiols located in the cytoplasmic glutamate oxidizing system, prior to the membrane-embedded NADH oxidation system. Inhibition by Cd2+ of delta mu (H+)-dependent ATP synthesis and of pH gradient (delta pH)-linked [14C]glutamate transport is a secondary effect due to cadmium-mediated inhibition of delta mu H+ generation at the cytoplasmic level. In washed cells of cadmium-resistant S. aureus 17810R oxidizing glutamate, Cd2+ accumulation was prevented due to activity of the plasmid-coded Cd2+ efflux system. Consequently, delta mu (H+)-producing and -requiring processes were not affected by Cd2+.

Intrinsic insensitivity to cadmium of the L-lactate oxidizing system in staphylococcoccus aureus.

Starved cells of cadmium-sensitive Staphylococcus aureus 17810S accumulated 109Cd via the Mn2+ porter energized by the membrane potential (delta psi) generated by L-lactate oxidation. However, Cd2+ accumulation did not result in inhibition of respiration and consequent generation of electrochemical proton gradient (delta muH+) via the respiratory chain. Thus, delta muH(+)-consuming processes, such as ATP synthesis and [14C]glutamate transport proceeded normally, despite the presence of Cd2+ in the cytoplasm. The mechanism of the intrinsic cadmium-insensitivity of the L-lactate oxidizing system is discussed.

Energetics of Cd2+ efflux system in cadmium-resistant Staphylococcus aureus 17810R.

Energetics of 109Cd efflux in resting cells of cadmium-resistant Staphylococcus aureus 17810R was assayed in 1 or 100 mM potassium/sodium phosphate buffer, pH7 (PB). Experiments with the use of inhibitors and ionophores showed that Cd2+ extrusion in this organism required ATP and either a pH gradient (delta pH) in 1 mM PB or membrane potential (delta psi) in 100 mM PB. The role of high phosphate ion concentration in delta psi-dependent Cd2+ efflux is discussed.

[Mesenteric cysts: rare cause of abdominal pain]. / Mesenterialzyste: seltene Ursache abdomineller Beschwerden.

For about two years a now 47-year-old woman had been suffering from heartburn, diarrhoea and constipation, at times also abdominal colic and a pressure sensation in the right lower abdomen. Physical examination and biochemical tests were according to age, except for an raised erythrocyte sedimentation rate of 25/50 mm. Ultrasound revealed an intraperitoneal echo-free multi-chambered structure, 11 x 5 x 2 cm, with dorsally increased echo density and smooth entry echo, apparently arising from the mesentery. Computed tomography confirmed these findings and at laparoscopy a cyst, 12 x 8 x 5 cm, was identified. It was filled with yellow clear fluid and arose from the mesentery of the ascending colon. The cyst was scooped out surgically. Histological examination showed a single-layered mesothelial coat. The postoperative course was unremarkable. At re-examination 8 months later the patient had remained symptom-free.

Anaerobic 109Cd accumulation by cadmium-resistant and -sensitive Staphylococcus aureus.

Washed cells of the cadmium-sensitive Staphylococcus aureus 17810S accumulated 109Cd under anaerobic conditions via the Mn2+ porter down delta psi in 1 or 100 mM phosphate buffer, pH 7; in washed cells of the cadmium-resistant S. aureus 17810R 109Cd accumulation was highly reduced. Nigericin did not stimulate anaerobic Cd2+ accumulation by strain 17810R in 100 mM phosphate buffer, suggesting that delta psi could energize Cd2+ efflux. In 1 mM phosphate buffer nigericin restored Cd2+ accumulation via the Mn2+ porter down delta psi in strain 17810R, indicating involvement of delta pH in Cd2+ extrusion. Increase of phosphate buffer concentration from 1 to 100 mM and addition of energy source at steady-state caused delta psi-dependent Cd2+ efflux from the nigericin-pretreated cells of strain 17810R. This suggests that the Cd2+ efflux system in S. aureus may require energy of both ATP and delta mu H+.