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Structure of the O-antigen of Escherichia coli 0119 lipopolysaccharide.

The structure of the O-polysaccharide component of the lipopolysaccharide produced by Escherichia coli 0119 was determined by the use of methylation analysis, periodate oxidation, 1D and 2D nuclear magnetic resonance spectroscopy, and mass spectrometric methods. The O-polysaccharide was found to be a high molecular weight polymer of a repeating pentasaccharide unit composed of D-mannose, D-galactose, L-rhamnose, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2,3-dideoxy-3-formamido-D-rhamnose residues (1:1:1:1:1) and had the structure: [formula: see text]

Bacterial Adhesion↗

Demethylation of the human telomerase catalytic subunit (hTERT) gene promoter reduced hTERT expression and telomerase activity and shortened telomeres.

Telomerase is the ribonucleoproteic complex involved in maintaining telomere size. It is expressed in germ and stem cells but not in normal somatic cells. In most tumors, telomerase is reactivated. In humans, telomerase activity is tightly regulated by expression of the hTERT gene. In a previous study, we found a direct correlation between methylation of the hTERT promoter and hTERT gene expression. In order to demonstrate this correlation, demethylation experiments were performed with the demethylating agent 5aza-2'-deoxycytidine (5azadC). Three telomerase-positive tumor cell lines (Lan-1, HeLa, and Co115), presenting a hypermethylated hTERT promoter, were treated with different doses and types of treatment for a long period. Analysis of methylation revealed a final hTERT promoter demethylation up to 95%. Quantification of hTERT mRNA showed that transcription was strongly repressed during drug exposure. In contrast, expression of c-Myc, an activator of hTERT promoter, was barely down-regulated or increased by the treatment. Using a TRAP assay, telomerase activity was semiquantified in all experiments. It strongly decreased or was suppressed after two to four passages. Finally, telomere length was measured by Southern blot. Their averages were not modified, but ranges concentrated around the mean. Thus, it is likely that hTERT promoter hypermethylation would be necessary for its expression.

Antimetabolites, Antineoplastic↗

Changes in the rhesus monkey's EEG responses to ethanol during chronic exposure.

This study was designated to document the changes in the CNS response to ethanol during chronic exposure in 5 male, 3.5--5.0 kg rhesus monkeys. Electrodes were implanted bilaterally into the amygdala, hippocampus and the calvarium over the frontal and temporal cortex. Ethanol or control solutions were administered intragastrically through indwelling cannulae. The 1.25 g/kg ethanol challenge dose was administered during EEG recordings. After one challenge dose, the animals received 60 days of chronic alcohol exposure (3.0 g/kg/day increasing to 8.0 g/kg/day). EEG was recorded every 10 days and analyzed by period analysis. Changes in the effect of the challenge dose were assessed by determining the percentage change of the EEG from pre-dose levels to selected times post-dose throughout the chronic alcohol exposure. The EEG response changed significantly during chronic alcohol treatment. Although each structure exhibited a slightly different pattern of change, the overall change was a shift from an excitatory response in the non-tolerant animal to an EEG slowing during chronic exposure. We suggest that such a change may be useful as a diagnostic marker for alcohol tolerance. In addition, the differential nature of the in vivo expression of alcohol tolerance in each brain area suggests that such analysis may provide a valuable tool for understanding the mechanism and expression of alcohol tolerance in the CNS.

Alcoholism↗

Structural features of a water soluble gum polysaccharide from Murraya paniculata fruits.

A water soluble gum polysaccharide was isolated from Murraya paniculata fruits. Hydrolytic experiments, methylation analysis, periodate oxidation studies and NMR data revealed that the polysaccharide was extensively branched and it consisted of 1,3-, and 1,3,6-linked beta-D-galactopyranosyl units, terminal beta-D-galactopyranosyl units and terminal alpha-D-glucopyranosyl 1,4-beta-D-galactopyranosyl units. Small amounts of 4-O-methylglucuronic acid residues were also present.

Anions↗

Losartan versus valsartan in the treatment of patients with mild to moderate essential hypertension: data from a multicenter, randomized, double-blind, 12-week trial.

BACKGROUND: Losartan, the first of the angiotensin II receptor blockers (ARBs) to be introduced, has been studied extensively in comparison with other classes of antihypertensive agents. Less research has been conducted on the efficacy and tolerability of losartan compared with that of other ARBs. OBJECTIVE: This randomized, multicenter, double-blind, parallel-group equivalence study was conducted to compare the antihypertensive efficacy and tolerability of a once-daily regimen of losartan with that of valsartan. METHODS: Patients > or = 21 years of age with mild to moderate hypertension, defined as a trough sitting diastolic blood pressure (SiDBP) between 95 and 115 mm Hg, were randomized to receive once-daily losartan (50 mg) or valsartan (80 mg) for 12 weeks. At the end of the sixth treatment week, patients in both groups with trough SiDBP > or = 90 mm Hg had their dose doubled for the remainder of the treatment period. Analysis of variance was used to compare treatment groups with respect to change in mean trough SiDBP from baseline to week 12. Within-treatment changes were analyzed using the paired t test. With at least 220 patients per treatment group, the study had 90% power to place a 90% CI on the difference between losartan and valsartan in SiDBP within the equivalence interval of +/- 2.5 mm Hg. RESULTS: A total of 495 patients were randomized, 247 to the losartan group and 248 to the valsartan group: 456 patients completed the study. Adjusted mean change from baseline values for trough SiDBP atthe end of 12 weeks of treatment were significantly different (P < 0.001) from zero in both the losartan group (-9.9 mm Hg) and the valsartan group (-10.1 mm Hg). At week 12, losartan was as effective as valsartan in lowering SiDBP, with a between-group difference of 0.2 mm Hg (90% CI, -1.3 to 1.7; P = 0.827). At week 6, the difference in SiDBP between groups was -1.3 mm Hg (90% CI, -2.7 to 0.0; P = 0.106). A similar pattern of results was obtained at weeks 6 and 12 for sitting systolic blood pressure. The percentage of patients reaching the SiDBP goal at week 6 (46% [112/2411 losartan; 42% [103/245] valsartan) and week 12 (57% [139/243] losartan; 59% [145/245] valsartan) was not significantly different between the treatment groups. Both losartan and valsartan were similarly well tolerated. Over the 12 weeks, the laboratory profiles of the 2 drugs were similar except for serum uric acid levels, which decreased from 6.0 to 5.7 mg/dL in the losartan group and increased from 5.9 to 6.0 mg/dL in the valsartan group (P = 0.001 for between-treatment difference). CONCLUSIONS: At starting and titrated doses, losartan and valsartan are similarly effective in reducing blood pressure in patients with mild to moderate hypertension. Losartan, but not valsartan, was associated with a decrease in serum uric acid levels.

Adult↗

A method for quantifying reflex responses from intra-muscular and surface electromyogram.

Measuring human reflex responses from electromyogram (EMG) traces in an accurate, repeatable and reliable way with a high degree of specificity has traditionally been a difficult task. This paper describes a new method that can be used to quantify reflex responses from both surface and intra-muscular EMG. This technique extends the classical cumulative sum (CUSUM) calculations by defining precise points for the calculation of latencies, durations and strengths to facilitate automatic reflex detection and permit the strength of a reflex to be defined in absolute units. The effect of varying the pre-stimulus time, the number of trials averaged and the amount of filtering used on the identification and classification of reflex parameters are also investigated. Furthermore, the effect of noise on these values, and how to remove it, is discussed. The new method, which is an expansion of the CUSUM analysis, is compared and contrasted with the more common threshold-crossing method in two different muscles: masseter and first dorsal interosseous (FDI), in experiments utilizing both mechanical and electrical stimulation. There are a number of advantages to using the new method; not only does the modified CUSUM method detect reflexes earlier than threshold-crossing methods but also the strength and duration are less susceptible to averaging and filtering parameters while giving a better indication of the reflex size. The data suggests that a pre-stimulus analysis period of at least 100 ms be used to correctly identify the variability inherent in EMG traces. It is also concluded that for subtle reflexes, 50 stimuli should be the minimum number used when spike trigger averaging is employed as lower numbers are associated with much greater pre-stimulus variability. Zero-phase filtering the rectified averaged EMG traces is recommended as this makes it easier to identify significant changes in the electrical activity of the muscle in question. In addition, noise estimation and removal from averaged rectified EMG recordings yields results that are a more accurate representation of the synaptic activity of the motor units in question.

Action Potentials↗

Circadian patterns of heart rate variability in normals, chronic stable angina and diabetes mellitus.

The purpose of our investigation was to compare circadian patterns of heart rate variability as assessed by 24-h ambulatory electrocardiographic (ECG) recordings in normal subjects, chronic stable angina, and Type 1 diabetes mellitus. The study population consisted of three groups: 12 normal subjects, 23 chronic angina patients, and 23 Type 1 diabetics. For purposes of analyzing circadian variation, the ECG recordings were divided into daytime (08:00-00.00 h) and night-time (00:00-08:00 h) periods. Analysis was performed for all time and frequency domain measures of heart rate variability, attempting to identify differences in day-to-night variability among these three groups. All time domain parameters except standard deviation of all 5-min mean RR intervals, and all frequency domain indices maintain significant circadian variations (P<0.0001), with the greatest day to night variation seen in normals, the least in diabetics, and intermediate values in chronic angina. These changes in heart rate variability circadian rhythms reflect significant reductions in cardiac parasympathetic activity with the most marked reduction in nocturnal vagal activity. Given the circadian pattern of myocardial ischemia and infarction, these data suggest that quantification of the magnitude of circadian variation in heart rate variability may have the potential to further risk stratify chronic angina and diabetes for future cardiac events.

Adult↗

Twentieth century overview of heavy metals in the Galician Rias (NW Iberian Peninsula).

The 18 Galician Rias, comprising 25% of the Iberian Peninsula coastline, form a unique system within the European continental Atlantic coast and provide a useful reference area for studies of metals. These rias are sensitive to anthropogenic influence due to the socioeconomic importance generated in these areas by industry, aquaculture, port activities, fishing, tourism and, accordingly, several islands lying within close proximity to the rias have recently been declared natural parks. Aquaculture in the Galician Rias is a growth industry and in 2000 the rias supported 3386 mussel rafts producing 2.5 x 10(8) kg year(-1), i.e. 40% of European Union total seafood production. The Galician Rias are partially stratified estuaries with positive residual circulation with fresh (runoff lower than 100 m3 s(-1) and salt water mixing inside the basins. This behaviour has direct consequences on metal fluxes whereby the rias behave as a barrier and accumulate metals in the sediments. Metals tend to deposit close to point sources and remobilization occurs inside the ria, leading to a seaward decrease in metal concentration. With the exception of the Ulla (Arosa Ria) and Eume (Ares-Betanzos Ria) head rivers and the Lagares river (Vigo Ria) the available data suggest that riverine freshwater inputs are not the main source of contamination. Metal studies reported in the literature mainly relate to Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn, and to a lesser extent Hg and Sn. Studies of other harmful metals such as Ag, Se and As are notably lacking. In addition, the studies have been localized and mainly restricted to the surface sediment. Data of heavy metals in the dissolved phase, suspended particulate matter and biota is scarce in the mainstream literature and should be considered in the future research. From the entire Galician coast, data from seven rias of socio-economic importance form the bulk of the published work. There is an urgent need to standardize procedures, employ 'clean' procedures to avoid contamination of samples and regular assessment of analytical accuracy with CRMs in order that results can be compared among the scientific community and produce reliable results. The continuation of coastal environmental studies requires an enlargement of the sampling locations as well as systematic and periodic analysis of known pollution sources in order to decide whether coastal regeneration action is needed. Studies dedicated to metals in the water column are lacking and need more attention in future research. The majority of the available data correspond to the fluvial front (30%) at ria head and, to a lesser extent, to the oceanic front (3%), under summer upwelling events. Current awareness of the rias suggests that they are not contaminated although there is some evidence of important localized anthropogenically induced enrichments at the outflow of the Lagares River (Pb, Zn), the San Simón inlet (Pb) and near to the harbor (Pb, Zn, Cu) in the Vigo Ria, the inner part (Cu, Zn, Hg, Pb) of the Pontevedra Ria, the Ulla River estuary (Cu, Cr, Mn, Ni; Zn) in the Arosa Ria, the inner zone (Cu, Pb, Zn) of the Coruña Ria, the Ferrol Ria (Zn) and the Eume Estuary (Zn, Mn, Ni, Co) in the Ares-Betanzos Ria. In addition, TBT enrichment has been found along the Galician coast, being highest in the industrial bays (Vigo, Coruña and Ferrol). Nevertheless, it is important to point out the lack of information concerning the relationship between metal concentrations in different fractions of sediments and/or water and their bioavailability and toxicity on biota in order to confirm metal contamination dangers. The present paper reviews all the information published on metals in the rias up to the year 2000 and takes a biogeochemical standpoint, i.e. metal presence in the ria reservoirs: water column, suspended particulate matter, sediments and biota, as well as the fluvial and oceanic frontiers.

Atlantic Ocean↗

Rusty peritoneal dialysis fluid after intravenous administration of iron dextran.

Rusty-colored peritoneal dialysate fluid was observed after intravenous administration of iron dextran to a patient with peritonitis being treated with vancomycin and rifampicin. The discoloration gradually cleared over a 24-hour period. Analysis of the fluid demonstrated that the discoloration could not be explained by the presence of erythrocytes or free hemoglobin. Iron (52 micrograms/dL) was detected in the fluid and decreased to undetectable levels as the discoloration cleared. Addition of iron dextran to an unused bag of peritoneal dialysis fluid to achieve an iron concentration of 52 micrograms/dL resulted in no discoloration. Addition of rifampicin at a clinically relevant serum concentration (10 micrograms/dL) to a different unused bag caused a light orange discoloration. Addition of iron dextran and rifampicin simultaneously in the concentrations mentioned to an unused bag caused a rusty discoloration almost as dark as that observed in our patient. We postulate, therefore, that a combination of iron and rifampicin caused the marked discoloration of our patient's peritoneal effluent.

Ascitic Fluid↗

Feasibility of fungi bioaugmentation in composting a flare pit soil.

The feasibility of fungi bioaugmentation in composting of a flare pit soil was studied in lab-scale composters. The preliminary screening tests, using a range of bulking agents and white rot fungi strains, were conducted to determine, best strain and bulking agent for the main experiments. The initial total petroleum hydrocarbon (TPH) of the flare pit soil was found to be 16%. The effect of moisture and bulking agent content and the fungi application on biodegradation of hydrocarbons were then evaluated based on a fractional factorial design over a 3-months period. Analysis of the TPH content of the soil after 98 days (using gravimetric method) showed an average of 29% reduction in most jars. Furthermore, gas chromatograph (GC) analysis of the oil extract from the samples showed 70-99% reduction in the peak area of the selected hydrocarbons. However, statistical analysis of the results did not show any significant effect due to the fungi application or the change in the moisture content (30-50% range). The results showed that the change in the bulking agent content was marginally significant for the hydrocarbon loss.

Biodegradation, Environmental↗

Introduction of an obstetric health information system: results of a pilot study in North Cameroon.

BACKGROUND: International safe motherhood programs have placed increasing emphasis on assessing progress in reducing maternal mortality in developing countries. We assess the feasibility and relevance of an obstetric health information system introduced in Maroua urban district in North Cameroon. METHODS: During the study period, an obstetric observation register was introduced for obstetric data collection, complemented by anthropological case studies on maternal deaths. RESULTS: At the end of the study period, implementation and data collection processes were correctly done, and the overall rate of completion of obstetric registers was 95% (ranging from 82.5% to 98.5% between maternity units). Eight hundred and twenty-six deliveries (n=826) were recorded and evenly distributed over the nine weeks of the study period. Eight women (1%) were transferred from non-surgical to surgical health facilities. Thirteen C-sections (n=13; 1.6%; CI: 0.8-2.7%) mainly in the provincial hospital of Maroua (11/13), and four maternal deaths were recorded, giving a maternal mortality rate of 4/826 (484 for 100,000; CI: 132-1240 for 100,000 deliveries). Nevertheless, anthropological enquiry recorded five maternal deaths during the same study period. Analysis of the geographical origin of these women showed that four of the five came from very remote areas. Rapid analysis and dissemination of results have initiated changes in obstetric practices (introduction of the partograph, modifications in the attitudes of health personnel), and also to the creation of a network between maternity units (those with and without surgical facilities) and provincial health headquarters. CONCLUSION: The introduction and use of a basic obstetric health information system combined with anthropological survey can provide a relatively accurate assessment of the maternal health situation. Such knowledge would be an excellent basis for implementing obstetric networking and relevant tools for active management of the obstetric pyramid at a regional level in developing countries.

Cameroon↗

An LC/MS/MS method for the quantitation of MTIC (5-(3-N-methyltriazen-1-yl)-imidazole-4-carboxamide), a bioconversion product of temozolomide, in rat and dog plasma.

A sensitive and selective HPLC/electrospray ionization tandem mass spectrometric (LC/ESI/MS/MS) method for the quantitative determination of MTIC (5-(3-N-methyltriazen-1-yl)-imidazole-4-carboxamide), a pharmacologically active hydrolysis product of temozolomide, was developed and validated over a linear range from 10 to 400 ng ml(-1) in dog plasma and from 10 to 500 ng ml(-1) in rat plasma. This HPLC method utilized small plasma volumes (70 microl), rapid sample processing, and isocratic elusion conditions to achieve sensitive and selective MS/MS detection. Samples were processed and analyzed one at a time every 4.5 min in order to compensate for the inherent instability of MTIC. Both MTIC and the internal standard DTIC [5-(3,3'-N,N'-dimethyltriazen-1-yl)-imidazole-4-carboxamide] were quantitated in the positive ion, selected reaction monitoring (SRM) mode. The lower limit of quantitation (LLOQ) was 10 ng ml(-1) in the plasma from both species. Inter-assay accuracy and precision of all calibration standards and quality control (QC) samples were within +/- 11 and 12%, respectively, with the exception of the LLOQ in rat plasma (17%). The validated method was used to determine the time dependent plasma concentration of MTIC in rats and dogs following a single oral dose of temozolomide. The standard curve and the quality control data indicate that the method performed acceptably throughout the sample analysis period.

Animals↗

Phenotypic antimicrobial resistance patterns in Pseudomonas aeruginosa and Acinetobacter: results of a Multicenter Intensive Care Unit Surveillance Study, 1995-2000.

Susceptibility data from the Intensive Care Unit (ICU) Surveillance Study for 10,361 isolates of Pseudomonas aeruginosa and 2,573 isolates of Acinetobacter tested at centers in the United States during 1995 to 2000 were analyzed. In all years, amikacin was the most active antimicrobial agent against P. aeruginosa, and imipenem was the most active agent against Acinetobacter. Resistance of both organisms to common therapeutic agents tested throughout the analysis period increased from 1995 to 2000, although the increase was not consistent for all drugs from year to year. The increases were higher among Acinetobacter, and for both organisms, the increase in resistance was greatest for ciprofloxacin. Among all P. aeruginosa tested in 1999 and 2000, resistance to ciprofloxacin was 9-11% higher for isolates from patients on general hospital wards than those from ICUs. Of the 3424 ICU isolates of P. aeruginosa tested in 1999 and 2000, 77 (2.2%) were multidrug-resistant (i.e., resistant to piperacillin, ceftazidime, imipenem, and gentamicin). Twenty (3.9%) isolates of Acinetobacter in 1999 and seven (1.9%) in 2000 were resistant to imipenem, ceftazidime, piperacillin-tazobactam, ciprofloxacin, and amikacin. Although resistance in both P. aeruginosa and Acinetobacter increased, multidrug-resistant (to > or =4 agents) strains were uncommon.

Acinetobacter↗

[Use of Diprivan in allergic patient].

Propofol may be indicated in patients with a history of allergy provided some precautions are taken. At present, the only contra-indication for propofol are patients with a known allergy to muscle relaxants. Any anaphylactoid reaction has to be systematically reported (e.g. to the Drug Safety Department of Zeneca Pharma) and the patient tested according to the standard protocol, in order to further disclose the underlying mechanisms and evaluate the risk factors. Periodical analysis of notifications of adverse reactions will facilitate regular epidemiological studies and the update of the information on the drug for the anaesthetic community.

Anaphylaxis↗

Development of a lethal Shiga toxin-producing Escherichia coli-infection mouse model using multiple mitomycin C treatment.

The aim of this study was to develop a lethal Shiga toxin-producing Escherichia coli (STEC) infection model in mice. A small inoculum of 5 x 10(3) CFU of STEC strain 89020087 to mice treated with streptomycin sulfate in drinking water (5 mg/ml) lead to marked increase in the excretion of the bacteria of up to 10(9)CFU/g feces within 18 h after the challenge. Combination of administration of 5 x 10(3) CFU of STEC followed by mitomycin C (MMC) treatment during the late log phase to the early stationary phase of STEC growth in the intestine lead to fatal infection. Periodic analysis showed that there is transient but dramatic increase in the Stxs (Stx1 and Stx2) concentration in the lower intestines after multiple MMC treatment. Histopathological analysis and blood chemistry revealed damages in both kidney and hematopoietic organs but not in the brain. Comparison of the virulence of 11 different STEC strains revealed that only strains which produced high amount of Stx2 responding to MMC treatment and exerted lethal toxicity to mice, suggesting that Stx2 plays a pivotal role in the lethal infection of STEC in the mouse model.

Alkylating Agents↗

A randomized, controlled trial of intravenous clodronate in patients with metastatic bone disease and pain.

To evaluate the effectiveness of intravenous clodronate in ameliorating refractory bone pain in patients with metastatic bone disease, 60 patients with established osseous metastases and persistent bone pain were randomized to receive either clodronate (600 mg or 1500 mg in 500 mL of normal saline) or 500 mL of saline as placebo. After 2 weeks, the patients were crossed over to receive the alternate treatment. After another 2 weeks, each patient and investigator made a blinded choice. Daily visual analogue scales (VAS) and analgesic diaries were recorded throughout the study period. Forty-six patients were evaluable (77%). A treatment x period interaction was identified in the VAS and daily morphine equivalent dose (DMED) scores. First period analysis of the VAS scores for general pain, pain at rest, and pain upon movement demonstrated an average reduction of 13, 14, and 24 mm, respectively, from baseline, but were not significantly different from changes following placebo. The average change in DMED was -6.4 (SE = 2.9) following clodronate and was +24.6 (SE = 14.9) following placebo (p = 0.03). In the blinded choice of which agent resulted in improvement in pain, 26 (57%) patients chose clodronate, 12 (26%) chose placebo, and eight (17%) had no preference (p = 0.0021). For the investigators who also made a blinded selection, clodronate was chosen in 30 (65%) patients, placebo in ten (22%) patients, and no difference was apparent in six (13%) (p < 0.0001). Intravenous clodronate appeared to have analgesic effect in patients with refractory bone pain due to metastatic bone disease. The optimal dose and duration of effect require further evaluation, particularly in patients with stable disease and persistent bone pain.

Adult↗

The Network Project: a multidisciplinary cancer education and training program in pain management, rehabilitation, and psychosocial issues.

Despite recent advances in cancer pain management and education, the undertreatment of cancer pain remains a national health problem. In response to the need for interdisciplinary education and training, we have developed "The Network Project" at Memorial Sloan-Kettering Cancer Center. This program includes a 2-week observership in cancer pain management, psychosocial oncology, and cancer rehabilitation. The efficacy of the Network Project Observership Program was evaluated with the use of a knowledge and attitude inventory which contains questions pertaining to cancer pain, psychosocial oncology, and cancer rehabilitation, and follow-up questionnaires completed one year after participation in the program. The knowledge and attitude inventory was completed by each participant both before and after the observership period. Analysis of the 152 observers who participated in the first 3 years of the Network Project's Observership Program demonstrated a significant improvement in knowledge of cancer pain, psychosocial issues, and rehabilitation issues (P < 0.0001). In addition, observers reported a significant increase in their local educational and training activities in the year following participation in the Network Project (P < 0.01). The Network Project, funded by the National Cancer Institute, is an interdisciplinary, multimodal training program which has been demonstrated to be an effective means for improving and disseminating knowledge regarding the multidisciplinary management of pain and psychological distress in cancer patients.

Community Networks↗

Deriving more up-to-date estimates of long-term patient survival.

Statistics reflective of patient survival should be as up-to-date as possible. We illustrate that traditional methods for analysing survival have important limitations with regard to up-to-dateness of long-term cumulative survival estimates. We propose an alternative approach, denoted "period analysis," that may be used to overcome or at least substantially reduce these limitations. The conceptual background and the computational realization of the new method are outlined and its application is illustrated by an analysis of long-term survival of patients with testicular cancer in Saarland, Germany.

Cohort Studies↗