A prospective study on Shiga toxin-producing Escherichia coli in children with diarrhea in Paraná State, Brazil.

AIMS: To examine stool specimens from children with diarrhea from Paraná State, southern Brazil, for presence of STEC. METHODS AND RESULTS: A PCR screening assay for stx genes was used to examine a loopful of confluent colonies of 306 stool samples cultures. In six (1.96%) of them, DNA fragments of the expected size were observed, and the presence of stx was confirmed by DNA sequencing. Then up to 100 single colonies from each of the six stool cultures were analyzed using the same PCR protocol. However, stx-positive colonies were found only in two of the cultures. The E. coli strains belonged to serotypes O69:H11 and O178:H19, and presented genotypes stx(1)eae ehxA and stx(1) respectively. Shiga toxin production was confirmed using the VTEC Screen Seiken. Except ampicillin, they were susceptible to all the antimicrobials tested. CONCLUSIONS: These results show that STEC may be an important cause of diarrhea in children of Paraná State, and that they are present in low numbers in stools. The strains belonged to serotypes not commonly found associated with STEC and probably present low virulence. SIGNIFICANCE AND IMPACT OF STUDY: These results indicate that molecular methods are required to diagnosis of STEC infections.

Revealing the faunal tapestry: co-evolution and historical biogeography of hosts and parasites in marine systems.

Parasites are integral components of marine ecosystems, a general observation accepted by parasitologists, but often considered of trifling significance to the broader community of zoologists. Parasites, however, represent elegant tools to explore the origins, distribution and maintenance of biodiversity. Among these diverse assemblages, host and geographic ranges described by various helminths are structured and historically constrained by genealogical and ecological associations that can be revealed and evaluated using phylogenetic methodologies within the context of frameworks and hypotheses for co-evolution and historical biogeography. Despite over 200 years of sporadic investigations of helminth systematics, knowledge of parasite faunal diversity in chondrichthyan and osteichthyan fishes, seabirds and marine mammals remains to be distilled into a coherent and comprehensive picture that can be assessed using phylogenetic approaches. Phylogenetic studies among complex host-parasite assemblages that encompass varying temporal and geographic scales are the critical context for elucidating biodiversity and faunal structure, and for identifying historical and contemporary determinants of ecological organization and biogeographic patterns across the marine biosphere. Insights from phylogenetic inference indicate (1) the great age of marine parasite faunas; (2) a significant role for colonization in diversification across a taxonomic continuum at deep and relatively recent temporal scales; and (3) a primary role for allopatric speciation. Integration of ecological and phylogenetic knowledge from the study of parasites is synergistic, contributing substantial insights into the history and maintenance of marine systems.

A method for measuring systolic and diastolic microcirculatory red cell flux within the canine myocardium.

BACKGROUND: Knowledge of the patterns of movement of red cells during the cardiac cycle in the microcirculation within the contracting myocardium is largely unknown. We describe a method of making such measurements in the canine myocardium using the technique of laser Doppler velocimetry. METHODS: A lensed 100 microm fiber-optic probe was inserted into the beating myocardium at various sites. Using an ultra-stable laser and achieving measurement stability by heterodyning the laser light and reflected light from the tissue, it was possible to obtain a stable high quality measurement of predominately red cell movement in the microcirculation. RESULTS: Unique regional patterns of red cell movement within the myocardium were observed. Epicardial flux was continuous with peaks while endocardial flux was predominately diastolic. Stopping flow in the epicardial artery for 5-6 s demonstrated that red cell movement continues in the microcirculation with some reduction followed by a delayed reactive hyperemia. Modeling demonstrates an important role for the small coronary veins in control of microcirculatory red cell movement. CONCLUSIONS: It is possible using laser Doppler velocimetry to measure red blood cell flux in the beating canine myocardium. Such measurements demonstrate a high degree of complexity which is not reflected in epicardial coronary arterial or venous flow.

Control of nitrogenase reactivation by the GlnZ protein in Azospirillum brasilense.

The glnZ mutant of Azospirillum brasilense (strain 7611) showed only partial recovery (20 to 40%) after 80 min of ammonia-induced nitrogenase switch-off, whereas the wild type recovered totally within 10 min. In contrast, the two strains showed identical anoxic-induced switch-on/switch-off, indicating no cross talk between the two reactivation mechanisms.

Recent developments in the structural organization and regulation of nitrogen fixation genes in Herbaspirillum seropedicae.

Herbaspirillum seropedicae is a nitrogen-fixing bacterium found in association with economically important gramineae. Regulation of nitrogen fixation involves the transcriptional activator NifA protein. The regulation of NifA protein and its truncated mutant proteins is described and compared with that of other nitrogen fixation bacteria. Nitrogen fixation control in H. seropedicae, of the beta-subgroup of Proteobacteria, has regulatory features in common with Klebsiella pneumoniae, of the gamma-subgroup, at the level of nifA expression and with rhizobia and Azospirillum brasilense, of the alpha-subgroup, at the level of control of NifA by oxygen.

Small bowel obstruction from internal hernia as a complication of colonoscopy.

After colonoscopy with polypectomy, a patient developed a surgically acute abdomen. Although abdominal radiology did not show free air, a presumptive diagnosis of bowel perforation was made and laparotomy performed. At the time of surgery the colon was normal, and there was no peritoneal contamination. A loop of ileum was discovered incarcerated into an internal paracecal hernia. The bowel was freed, and the operation was completed without need for resection. Several cases of incarcerated inguinal hernia resulting from endoscopy are described in the medical literature; this is the first reported case of an incarcerated internal hernia as a complication of colonoscopy.

A method for detecting chaos in canine myocardial microcirculatory red cell flux.

OBJECTIVE: To determine whether red cell movement, as measured by laser Doppler velocimetry, in the capillary net of the beating heart is chaotic. METHODS: Using two dog hearts, in situ red blood cell flux was measured at many sites. Simultaneously, epicardial arterial flow and left ventricular pressure were recorded via transit-time flowmeter and catheter manometer, respectively. The presence or absence of chaos was tested by two methods: Lyapunov exponents and correlation dimension. RESULTS: For capillary red cell flux, the Lyapunov was strongly positive at most sites. It was less so for coronary arterial flow and least for left ventricular pressure. Correlation dimension calculation was less able to distinguish the presence or absence of chaos in capillary red cell tissue flux, coronary arterial flow, and left ventricular pressure. CONCLUSIONS: Capillary red cell flux (movement of red cells in capillaries) is nonlinear, (i.e., chaotic). This complexity suggests that the primary control for oxygen delivery to cardiac myocytes by red blood cells resides in the microcirculation. Also, capillary red cell flux is bifractal, suggesting an ordering of control.

Calcified subcutaneous arterioles with infarcts of the subcutis and skin ("calciphylaxis") in chronic renal failure.

Patients with chronic renal failure (CRF) are at increased risk for pathological calcifications because of increased serum calcium-phosphorus products. A minority, including those undergoing dialysis, develop a syndrome of deep skin ulcerations in association with calcification of subcutaneous arterioles. The body distribution of the skin lesions may be proximal (central), distal (peripheral), or both. Since 1968, this syndrome has been called "calciphylaxis" in the belief that it is the human analogue of Selye's experimental models of tissue calcification. Our review emphasizes that this syndrome comprises two separate processes not found in calciphylaxis: calcification of subcutaneous arterioles and infarctions of subcutaneous adipose tissue (panniculus adiposus) and skin. The infarctions are acute and clinically dramatic, whereas the calcific arteriolopathy is preexistent, having developed slowly, sometimes over years, and silently. Separating these two processes facilitates analyses of pathogenetic factors, such as those that target subcutaneous arterioles for calcification and those that interfere with blood flow through the calcified arterioles, sufficient in some patients to cause the infarctions, and of why obesity in CRF is a syndrome risk factor. This approach further helps to provide a much needed standardized definition of the syndrome, thereby facilitating comparisons of the results of such treatments as parathyroidectomy, anticoagulants, and phosphate binders. Finally, the separation shows why the application of such terms as calciphylaxis and calcifying panniculitis to this syndrome is inappropriate.

Sequencing and functional analysis of the nifENXorf1orf2 gene cluster of Herbaspirillum seropedicae.

A 5.1-kb DNA fragment from the nifHDK region of H. seropedicae was isolated and sequenced. Sequence analysis showed the presence of nifENXorf1orf2 but nifTY were not present. No nif or consensus promoter was identified. Furthermore, orf1 expression occurred only under nitrogen-fixing conditions and no promoter activity was detected between nifK and nifE, suggesting that these genes are expressed from the upstream nifH promoter and are parts of a unique nif operon. Mutagenesis studies indicate that nifN was essential for nitrogenase activity whereas nifXorf1orf2 were not. High homology between the C-terminal region of the NifX and NifB proteins from H. seropedicae was observed. Since the NifX and NifY proteins are important for FeMo cofactor (FeMoco) synthesis, we propose that alternative proteins with similar activities exist in H. seropedicae.

Pharmacokinetics and hemodynamic effects of diltiazem in healthy volunteers: comparing resting with the effect of exercise.

PURPOSE: To determine the steady-state plasma concentrations of diltiazem (DTZ) and hemodynamic effect in humans at rest and during exercise. METHODS: Healthy volunteers (10 F, mean age 22, and 11 M, mean age 24) were recruited. Prior to receiving DTZ, each volunteer performed two 3-minute stages of treadmill exercise according to the Bruce protocol. Intra-arterial BP and ECG recordings were obtained before, during and immediately post exercise. Each volunteer then received DTZ 60 mg qid for one week. The same exercise protocol was repeated 1 h after the last dose. Steady-state plasma concentrations of DTZ were determined by a previously reported HPLC. RESULTS: DTZ decreased resting DBP from 84 +/- 13 to 79 +/- 10 mmHg (p > 0.05), and HR from 89 +/- 11 to 82 +/- 13 bpm (p < 0.05). During exercise, an average of 32 and 10% increase in SBP and DBP, respectively, and a 47% increase of HR was found (p < 0.05). DTZ limited these increases to 21% for SBP, 5% for DBP, and 44% for HR (p < 0.05 for drug effect). Steady-state plasma DTZ concentrations were 141 +/- 56 ng/ml. CONCLUSION: DTZ significantly decreased resting HR but not BP in health volunteers. It decreased both hemodynamic variables during exercise. Thus, the hemodynamic effects of diltiazem are more profound during exercise, and may be more useful surrogate markers for calcium antagonists and other cardiovascular agents in healthy volunteer studies.

The coronary circulation: quo vadis?

This review examines past and present observations concerning the structure and function of the coronary circulation in health and disease. These observations are considered in the context of how we might proceed to increase understanding of this circulation in the future. The coronary microcirculation with its intimate relationship of capillaries and myocytes and the coronary venous circulation are identified as parts of the coronary circulation in need of further study. Observations using a laser Doppler velocimeter placed within the beating myocardium are presented. Such devices measure the velocity of red cells continuously (red cell flux) and can demonstrate that tissue hematocrit and hence oxygen delivery can be regulated independently of total epicardial arterial volume flow. Implications for the understanding of the pathophysiology of coronary artery disease are presented. As considerations are given to molecular genetic techniques to revascularize the ischemic myocardium we will require a more complete knowledge of coronary circulatory dynamics, myocardial support tissues' responses, and cardiac myocyte interactions to design appropriate interventions. The clinical trial is an appropriate clinical tool to measure effectiveness but a blunt instrument to determine pathophysiology. The purpose of the review is to suggest that advances in measurement of end points are required to permit the right question to be posed.

Outcomes of coronary artery bypass surgery in elderly people.

OBJECTIVES: To document the impact of coronary artery bypass (CABG) surgery on quality of life in elderly people three months after surgery; to compare quality of life measures; to examine predictors of poor quality of life; and to assess the predictive validity of global clinical rating scales. STUDY DESIGN: Prospective cohort study with postoperative, three-month and one-year follow-up. SETTING: A 700-bed tertiary care teaching hospital in Halifax, Nova Scotia. PATIENTS: One hundred of 200 consecutive patients, 75 years and older, undergoing CABG. MEASURES: Demographic information, clinical data and global clinical ratings were collected at baseline to assess risk for adverse outcome (death, stroke, functional impairment) at each follow-up. Quality of life three months post-CABG was documented using the RAND 36-Item Health Survey and the Seattle Angina Questionnaire. MAIN RESULTS: Perioperative deaths occurred in four patients, disabling strokes in six and postoperative complications in seven. Three additional deaths occurred by the three-month follow-up and nine more by one year. On average, important improvements in quality of life were observed at three months' follow-up, using both assessments of quality of life. CONCLUSIONS: Many elderly patients do well after CABG surgery, but a higher proportion than is seen in younger patients have complications leading to death and disability. A global clinical measure by the attending cardiologist at baseline did not identify patients at a higher risk. Follow-up studies are needed to assess long term outcomes of older patients at higher risk of death or a poor quality of life after CABG surgery.

Brain-derived HIV-1 tat sequences from AIDS patients with dementia show increased molecular heterogeneity.

HIV-1 infection results in a dementing illness affecting 20% of patients with AIDS. Several HIV-1 genes have been implicated in the pathogenesis of HIV-induced neurological disease. To search for distinct HIV-1 sequences associated with the development of dementia, brain-derived tat, env, and pol sequences were examined from AIDS patients defined pre-mortem as demented (HIV-D)[n=5] or non-demented (HIV-ND)[n=5]. Estimations of evolutionary distances and frequency of non-synonymous mutation rates revealed significant differences between brain-derived tat, env, and pol-encoded reverse transcriptase sequences. However, established zidovudine-associated resistance mutations in reverse transcriptase sequences were identified in only one HIV-D and one HIV-ND patient despite prolonged treatment of some patients. Non-synonymous/synonymous substitution rates among the tat sequences derived from patients with HIV-D were significantly higher compared to the HIV-ND group (P < 0.001). The ratios of transversions to transitions were also significantly higher among the HIV-D tat sequences (P< 0.01). Phylogenetic analyses showed clustering of sequences from each clinical group among the brain-derived tat and env sequences. These studies indicated that differing selective forces act on individual HIV-1 genes in the brain which may influence the development of dementia.

Phylogenetic relationships among prokaryotic and eukaryotic catalases.

Seventy-four catalase protein sequences, including 29 bacterial, 8 fungal, 7 animal, and 30 plant sequences, were compiled, and 70 were used for phylogenetic reconstruction. The core of the resulting tree revealed unique, separate groups of plant and animal catalases, two groups of fungal catalases, and three groups of bacterial catalases. The only overlap of kingdoms occurred within one branch and involved fungal and bacterial large-subunit enzymes. The other fungal branch was closely linked to the group of animal enzymes. Group I bacterial catalases were more closely related to the plant enzymes and contained such diverse taxa as the Gram-positive Listeria seeligeri, Deinocococcus radiodurans, and gamma-proteobacteria. Group III bacterial sequences were more closely related to fungal and animal sequences and included enzymes from a broad range of bacteria including high- and low-GC Gram positives, proteobacteria, and a bacteroides species. Group II was composed of large-subunit catalases from diverse sources including Gram positives (low-GC Bacilli and high-GC Mycobacteria), proteobacteria, and species of the filamentous fungus Aspergillus. These data can be interpreted in terms of two gene duplication events that produced a minimum of three catalase gene family members that subsequently evolved in response to environmental demands. Horizontal gene transfer may have been responsible for the group II mixture of bacterial and fungal large-subunit catalases.

Decreasing mortality from acute myocardial infarctions: effect of attack rates and case severity.

Mortality from myocardial infarction (MI) has declined in many countries and the reasons for the decline have not been fully quantified. We used the database of the Halifax County MONICA Project to test the hypothesis that the decline of in-hospital mortality from MI can be explained by a trend toward less severe disease as opposed to improved treatment. During the study period 1984-1993, 14,130 people aged 25-74 had been admitted to hospital with suspected MI. Of these, 3774 were diagnosed as definite MI by standardized criteria (480 fatal). For each patient, clinical history, serial cardiac enzymes, and ECG treatment regimen during hospital stay were extracted from patient charts. Survival status 28 days after onset of symptoms was determined. A severity index predicting 28-day case fatality was derived from health status at admission time. During the study period the rate of definite MI in the MONICA target population showed a general downward trend from 221 to 179 per 100,000/year (p = 0.0002). The severity index increased during the observation time (p < 0.0001), predicting 25% higher mortality. Case fatality fluctuated, but showed a marginally significant decline. We conclude that part of the decreased in-hospital mortality from MI is due to lower attack rates. The remainder occurred despite increased case severity and is possibly due to improved in-hospital treatment.

Red cell flux during the cardiac cycle in the rabbit myocardial microcirculation.

OBJECTIVE: (1) To measure regional phasic myocardial red cell flux during a cardiac cycle using a laser Doppler velocimeter. (2) To test the responses of regional red cell flux to a vasodilator (adenosine), a vasoconstrictor (angiotensin II), and an inotrope (isoproterenol). METHODS: Using an anaesthetised open-chest rabbit with the pericardium intact a 140-micron-tip fibre optic probe was placed in the left ventricular myocardium in various locations. With the fibre in place drugs were given to alter myocardial loading conditions while red cell flux was registered. RESULTS: Phasic red cell flux was similar in the epicardium to endocardium giving an average endo/epi ratio of 1.14 in the rabbit heart. At least two peaks of increased red cell flux within a single cardiac cycle were observed. Some unique patterns for red cell flux were observed in specialised myocardial structures. Adenosine increased red cell flux but minimally changed the pattern of phasic flux throughout the cycle. CONCLUSIONS: Laser Doppler velocimetry permits the recording of phasic red cell flux during the cardiac cycle in the myocardial microcirculation. Its pattern is determined by both coronary arterial inflow and venous outflow. The pattern of red cell flux may be characteristic for a region-probably determined by difference in tissue pressure (attributable to the pattern of muscle fibre shortening and collagen tethering) and changes in capillary length and density.

Effect of diltiazem on plasma concentrations of oxypurines and uric acid.

To determine the clinical effect of diltiazem on the metabolism of adenosine, and its importance in ischemic heart disease, arterial plasma concentrations of the purine metabolites were determined in 21 healthy volunteers (10 female and 11 male) and 19 patients with effort angina (8 female and 11 male) before, during, and immediately after standard treadmill exercise tests conducted before and after they had taken 60 mg diltiazem (Cardizem; Hoechst Marion Roussel, Laval, QC, Canada) four times a day for 1 week. The results showed that the cardiac patients had significantly lower mean plasma concentrations of uric acid (46.82 +/- 25.51 versus 95.47 +/- 35.41 micrograms/ml, p 0.05), inosine (0.25 +/- 0.19 versus 0.84 +/- 0.17 microgram/ml, p < 0.05), and hypoxanthine (0.28 +/- 0.35 versus 0.50 +/- 0.27 microgram/ml, p < 0.05). Diltiazem decreased the mean resting plasma concentrations of uric acid in patients (uric acid 43.47 +/- 22.26 versus 46.82 +/- 25.51 micrograms/ml, p < 0.05) and healthy volunteers (uric acid 85.68 +/- 26.71 versus 95.47 +/- 35.41 micrograms/ml, p < 0.05). There was no statistically significant change in the plasma concentrations of the purine metabolites during exercise (p < 0.05). Female subjects had significantly lower plasma concentrations of uric acid than males (patients, 34.87 +/- 26.93 versus 55.78 +/- 21.25 micrograms/ml; healthy volunteers, 84.79 +/- 32.07 versus 104.22 +/- 37.05 micrograms/ml; p < 0.05 for both). Results of the study suggest that normal therapeutic doses of diltiazem may modulate the metabolism of adenosine and that some of the purine metabolites may be useful markers for specific types of ischemic heart disease.

Effect of diltiazem on intraarterial blood pressure and heart rate during stress testing in patients with angina: a gender comparison study.

The purpose of this study was to measure the blood pressure and electrocardiographic responses of a small, matched group of women (n = 8) and men (n = 9) who experienced typical, effort angina during an exercise on the treadmill (up to the second stage of a Bruce protocol). These responses were measured before and after therapy with diltiazem (60 mg four times daily for 1 week). Reports of previous studies have described significant gender differences in blood pressure responses to diltiazem in healthy volunteers tested with the same protocol. In contrast to the data in healthy individuals, gender differences in blood pressure responses to exercise before and after diltiazem administration were not observed. Results of analysis of pulse pressure responses to exercise were also similar in male and female patients with angina. A significant postexercise drop in blood pressure was observed, which was augmented by diltiazem. These data suggest that gender differences in drug action may be difficult to demonstrate in patients with vascular disease.

Stress, coping, and social support as psychosocial factors in readmissions for ischaemic heart disease.

This exploratory investigation aimed to identify psychological and social factors associated with early (4 months postdischarge) readmission to hospital of persons with ischaemic heart disease (IHD). Data on stress (Stress Appraisal Scale), coping strategies (Ways of Coping Questionnaire), and social network/social support (Norbeck Social Support Questionnaire) were collected from 100 persons with the primary diagnosis of IHD prior to discharge from hospital. Twenty-four persons were readmitted during the 4-month study period. The sample was divided into two groups for analysis: (1) persons with a first admission for IHD, and (2) persons with a prior history of admission for IHD. In comparison to persons with multiple admissions for cardiac illness, first time admissions appraised their IHD as less central and threatening, perceived less affective and affirmational support from health professionals, and identified health professionals less and neighbours more as sources of support. Greater use of the coping strategy 'seeking social support' was associated with the readmission of persons who had had their first admission for IHD. Less use of the coping strategy 'accepting responsibility' was associated with the readmission of persons who had a history of prior admission for IHD.

A tandem array of 5S ribosomal RNA genes in Pythium irregulare.

The 5S ribosomal RNA genes of the oomycete Pythium irregulare exist in tandem arrays unlinked to the rDNA repeat unit. A clone with a 9.2-kb insert containing an array of 5S genes was identified in a lambda genomic library and was characterized by restriction mapping and partial sequencing. The array consisted of 9 apparently identical 5S genes and their spacers in tandem, followed by a diverged 5S-like sequence that is likely to be a pseudogene. This gene arrangement, although almost universal in plants and animals, is rare in fungi and protists.