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PubMed1992

DNA probe for strain typing of Cryptococcus neoformans.

A 7-kb linear plasmid, harbored by a URA5 transformant, hybridized to all the chromosomes of Cryptococcus neoformans separated by contour-clamped homogeneous electric field electrophoresis. Its linear maintenance was determined to have been facilitated by the presence of telomere-like sequences at its free ends. Hybridization of this plasmid to AccI-digested genomic DNAs of 26 C. neoformans strains generated 21 unique DNA fingerprints. The DNA fingerprints of isolates within the same serotype were more similar to one another than to those from different serotypes. An acapsular clinical isolate, strain 602, widely used in immunological studies and previously thought to be in serotype D, showed DNA fingerprints typical of serotype A isolates. Isogenic strains of C. neoformans exhibited DNA fingerprints that were identical to one another. The DNA fingerprints were stable and reproducible in spite of repeated transfers in the laboratory on either complex (1% yeast extract, 2% Bacto Peptone, 2% glucose) or minimal (yeast nitrogen base) medium. The DNA fingerprints of isolates recovered from primary blood and cerebrospinal fluid cultures of patients for whom AIDS had been diagnosed showed that the original infection in each of these patients contained a homogeneous population of C. neoformans. The DNA fingerprints of isolates recovered from different tissues of infected mice and from patients undergoing different drug therapy regimens were also found to be very stable.

AIDS-Related Opportunistic Infections↗
PubMed2005

Pilot study to evaluate microarray hybridization as a tool for Salmonella enterica serovar Typhimurium strain differentiation.

In developed countries, Salmonella enterica subspecies 1 serovars Enteritidis and Typhimurium range among the most common causes of bacterial food-borne infections. The surveillance and typing of epidemic Salmonella strains are important tools in epidemiology. Usually, Salmonella enterica subspecies 1 serovars are differentiated by serotyping for diagnostic purposes. Further differentiation is done by phage typing as well as molecular typing techniques. Here we have designed and evaluated a prototype DNA microarray as a tool for serovar Typhimurium strain differentiation. It harbors 83 serovar Typhimurium probes obtained by differential subtractive hybridization and from the public database. The microarray yielded reproducible hybridization patterns in repeated hybridizations with chromosomal DNA of the same strain and could differentiate five serovar Typhimurium reference strains (DT204, DT104, DT208, DT36, and LT2). Furthermore, the microarray identified two distinct groups among 13 serovar Typhimurium DT104 strains. This correlated with observations from pulsed-field gel electrophoresis analysis. Twenty-three further serovar Typhimurium strains were analyzed to explore future directions for optimization of the simple 83-probe DNA microarray. The data presented here demonstrate that DNA microarrays harboring small numbers of selected probes are promising tools for serovar Typhimurium strain typing.

Bacterial Typing Techniques↗
PubMed1970

A study of abnormal muscular patterns in diverticular disease of the colon using the polysiloxane foam enema.

Three-dimensional, functional studies of the left colon using a polysiloxane foam enema have shown that three types of impressions due to muscular contractions are commonly found in the region of the rectum and sigmoid, and that at least two of these are closely associated with the presence of diverticular disease. Each type of abnormality has been defined and its extent and frequency assessed. A tentative mechanism for the development of the main muscular abnormality through muscle thickening and simple sigmoid curves is put forward, and it is suggested that the third type of abnormality may be associated with sphincteric action at the recto-sigmoid junction. The reproducibility of findings on repeated examinations must mean that there is a local anatomical basis for the muscular impressions seen and that such contractions do not represent evanescent peristaltic type activity. Sigmoid resection and myotomy as practised for diverticular disease should eliminate the first type of abnormality in the majority of cases, but it is necessary to carry the operation well distally to ensure this. Neither operation is likely to correct the type III abnormality, and further work is necessary to determine whether this is responsible for some of the symptoms persisting after surgery.

Colon↗
PubMed1997

Genotypic diagnosis of familial Mediterranean fever (FMF) using new microsatellite markers: example of two extensive non-Ashkenazi Jewish pedigrees.

Familial Mediterranean fever is an autosomal recessive disease characterised by multiple attacks of serosal inflammation in the absence of treatment. In the absence of timely diagnosis, renal amyloidosis is a life threatening complication. The diagnosis is often missed because no specific test is available. Early colchicine treatment prevents attacks and renal complications. The FMF gene (MEF) has been mapped to chromosome 16p 13.3 but has not yet been identified. We compared the suitability of a series of microsatellite markers (four of them were new) and propose the routine use of seven of these markers, exhibiting alleles in strong linkage disequilibrium with the disease and informative in 100% of diagnosed patients. Moreover, the discovery of a homozygous status for the 3-3-9 (or 3-3-18) haplotype at the core loci (D16S3070, D16S3082, and D16S3275), which was found in 73% non-Ashkenazi Jewish patients, points to a diagnosis of FMF, even in sporadic cases, with a risk of error of only 2.10(-5). Two extensive pedigrees covering most indications for genetic counselling are presented, showing that it is now possible both prospectively and retrospectively to identify members likely to have MEF mutations. With the help of this accurate test, colchicine treatment can be better targeted, especially where the symptomatology is mild or atypical.

Chromosome Mapping↗
PubMed2006

Spinal cord stimulation suppresses bradycardias and atrial tachyarrhythmias induced by mediastinal nerve stimulation in dogs.

Spinal cord stimulation (SCS) applied to the dorsal aspect of the cranial thoracic cord imparts cardioprotection under conditions of neuronally dependent cardiac stress. This study investigated whether neuronally induced atrial arrhythmias can be modulated by SCS. In 16 anesthetized dogs with intact stellate ganglia and in five with bilateral stellectomy, trains of five electrical stimuli were delivered during the atrial refractory period to right- or left-sided mediastinal nerves for up to 20 s before and after SCS (20 min). Recordings were obtained from 191 biatrial epicardial sites. Before SCS (11 animals), mediastinal nerve stimulation initiated bradycardia alone (12 nerve sites), bradycardia followed by tachyarrhythmia/fibrillation (50 sites), as well as tachyarrhythmia/fibrillation without a preceding bradycardia (21 sites). After SCS, the number of responsive sites inducing bradycardia was reduced by 25% (62 to 47 sites), and the cycle length prolongation in residual bradycardias was reduced. The number of responsive sites inducing tachyarrhythmia was reduced by 60% (71 to 29 sites). Once elicited, residual tachyarrhythmias arose from similar epicardial foci, displaying similar dynamics (cycle length) as in control states. In the absence of SCS, bradycardias and tachyarrhythmias induced by repeat nerve stimulation were reproducible (five additional animals). After bilateral stellectomy, SCS no longer influenced neuronal induction of bradycardia and atrial tachyarrhythmias. These data indicate that SCS obtunds the induction of atrial arrhythmias resulting from excessive activation of intrinsic cardiac neurons and that such protective effects depend on the integrity of nerves coursing via the subclavian ansae and stellate ganglia.

Animals↗
PubMed1980

Physiological and pharmacological properties of canine trachealis muscle in vivo.

In 23 dogs anesthetized with chloralose and urethan, we developed a preparation that premitted measurement of isometric tension in segments of the tracheal posterior membrane in situ. Length-tension studies showed an optimal length for contraction in each of 5 segments; in all 5 segments, length was optimal when resting tension was 22-35 g/cm. In 8 other segments, stimuli were delivered by systemic intravenous injections, by electrical stimulation of nerves, or by selective injections into the tracheal circulation; the time between delivery of repeated stimuli required for reproducible responses differed for each form of stimulation. Hypoxemia increased tension by 23.1 +/- 5.5 (SE) g/cm in 10 out of 10 other segments; these increases were the results of reflexes, not direct effects. Because the innervation and vascular supply of these segments were largely intact, we concluded that this preparation permits the study of neurohumoral mechanisms which modulate smooth muscle contraction in a prototypic central airway in vivo.

Acetylcholine↗
PubMed1994

Mechanosensitive properties of pelvic nerve afferent fibers innervating the urinary bladder of the rat.

1. Single-unit activity was recorded from S1 dorsal root afferent fibers in pentobarbital-anesthetized rats. In 25 experiments, 245 afferent fibers were identified by electrical stimulation of the pelvic nerve. Fifty-two percent were C fibers (conduction velocity: 1.70 +/- 0.04 m/s; mean +/- SE) and 48% were A delta-fibers (conduction velocity: 11.07 +/- 0.95 m/s). 2. Of 245 pelvic nerve afferent fibers, 92 (38%) responded to noxious urinary bladder distension (UBD; 80 mmHg); 57 of these fibers were C fibers and 35 were A delta-fibers. Forty-four fibers responded to colorectal distension (CRD; 80 mmHg); 32 were C fibers and 12 were A delta-fibers. A total of 39 fibers were identified in the anal mucosa; 3 were C fibers and 36 were A delta-fibers. Seventy fibers (28%) in these experiments were unresponsive to either UBD or CRD or to probing of the anal mucosa; 32 were unmyelinated C fibers and 38 were A delta-fibers. 3. Reproducibility of responses to repeated UBD (80 mmHg, 20 s; 8 trials at 4-min intervals) was tested in 10 fibers. In nine fibers, responses to repeated distension did not change; one fiber exhibited a progressive decrease in response magnitude after the third trial. 4. Of the 92 afferent fibers that responded to UBD, 45 were further characterized for responses to graded intensities of UBD. Forty fibers had some resting activity (1.7 +/- 0.3 impulses/s) and five fibers exhibited no ongoing activity. The response to UBD adapted slowly during the 20-s period of phasic UBD or during slow isotonic filling of the bladder. 5. The stimulus-response function (SRF) of fibers (n = 45) to graded UBD was monotonic < or = 80 mmHg. Thresholds for responses were determined after extrapolation of the least-squares linear regression line to the ordinate, and varied between 0 and 45 mmHg. The frequency distribution profile of thresholds revealed two populations of pelvic nerve afferent fibers in the urinary bladder: a larger group (n = 36) of low-threshold (LT) fibers (5.7 +/- 1.0 mmHg) and a smaller group (n = 9) of high-threshold (HT) fibers (34 +/- 2.5 mmHg). 6. Responses of four LT fibers to graded UBD were tested before and 30 min after instillation of 0.5 ml of 0.5% acetic acid (pH 3) into the bladder. The mean threshold for response of these fibers before instillation of acetic acid (9.4 +/- 3.1 mmHg) more than doubled (to 22.3 +/- 6.7 mmHg) after instillation of acetic acid.(ABSTRACT TRUNCATED AT 400 WORDS)

Acid-Base Equilibrium↗
PubMed1994

Therapeutic splenic ultrasound for immune thrombocytopenia in children.

The spleen acts as a major site of clearance of antibody-coated platelets from circulation in immune thrombocytopenia (ITP). Splenectomy carries a high cure rate. The biological effect of a single therapeutic dose of ultrasound directed transthoracically to the spleen at 1 MHz and 1 W/cm2 with a mean treatment time of 5 min as generated by Sonopuls 463 (Enraf Nonius) was studied in 30 children with ITP (20 chronic, 10 acute) aged 8-14 years (median 10) and 10 control children. The chronic ITP cases had platelet counts (PC) of 20-50 x 10(9)/l (mean 36 x 10(9)/l), showed peak responses at 4 h after exposure 5-18 x 10(9)/l (mean 10 x 10(9)/l) in 70% of cases, while the remaining 6 patients showed either no change in PC (n = 3) or a decline in PC (n = 3) 5-7 x 10(9)/l. Children with acute ITP had pretreatment PC of 30-50 x 10(9)/l (mean 40 x 10(9)/l). All had increments of PC after ultrasonic exposure (10-30 x 10(9)/l; mean 18 x 10(9)/l) peaking at 4 h. Six patients with acute ITP maintained the rise in PC while in an other 4, PC returned to baseline in 24 h. The control group showed no change in PC. This therapy was well tolerated and was not associated with significant change in serum lactate dehydrogenase (LDH) levels except in 2 cases with chronic ITP, in whom the LDH levels doubled. All above results were reproduced when therapy was repeated 2 weeks later. In conclusion, this therapy would seem to be safe and well tolerated at such a dose. The effectiveness, rapidity and low cost of this therapy compared with conventional approaches may suggest its use as an alternative therapy in ITP.

Acute Disease↗
PubMed1994

Limited expansion of the (CAG)n repeat of the Huntington gene: a premutation (?).

Huntington's disease (HD) is an autosomal dominant disorder with choreic movements, psychiatric manifestations and cognitive dysfunction. Recently the IT15 gene on chromosome 4p has been identified containing an unstable and expanded trinucleotide repeat in patients with HD. We report on the characteristics of this repeat in 248 individuals from 41 Belgian HD families. The length of the expanded repeat was defined precisely and reproducibly on an ALF sequencer and correlated well with the age of onset (r = -0.72). Paternal transmission of the expanded repeat resulted on average in a significantly longer repeat length (+2.79 repeats) than maternal transmission (-0.29 repeats). (CAG)n repeat of a premutation (?) size was observed in this population with subsequent expansion in the disease range. Presymptomatic or prenatal testing using only linked markers may be problematic in these cases.

Adult↗
PubMed1999

Determination of tumour regression rates during radiotherapy for cervical carcinoma by serial MRI: comparison of two measurement techniques and examination of intraobserver and interobserver variability.

Tumour regression rates of 11 patients with cervical carcinoma were estimated during external beam radiotherapy (EBRT) using serial MRI (average time interval 7 days; range 3-15 days). An average of five investigations (range 4-8) was performed per subject. Tumour volume was measured by two observers using the Cavalieri method of modern design stereology in combination with (a) planimetry and (b) point counting. The mean precision of all the volume estimates obtained by manually tracing the outline of the tumour was 6.6%. The mean precision obtained by counting an average of 176 points per investigation on the same transects was 6.7%. The intraobserver repeatability of planimetry, interobserver reproducibility of planimetry and point counting were excellent with no significant difference between the volume estimates obtained using either technique. Based on the planimetry measurements, initial tumour volumes ranged from 6.5 to 222 cm3 (mean 63 cm3, median 44 cm3). Based on the point counting measurements, initial tumour volumes ranged from 7.2 to 235 cm3 (mean 68 cm3, median 46 cm3). Tumour regression began within a few days of commencing EBRT and showed an exponential relationship with time (p < 0.01). There was good agreement between the regression rates obtained by planimetry and those obtained by point counting. No significant correlation was found between initial tumour volume and tumour regression rate for either planimetry or point counting. Planimetry measurements were, on average, obtained in about half the time taken for point counting (i.e. 30 min and 50 min, respectively). Although point counting is generally likely to be the more efficient approach, planimetry may be the preferred approach for estimating tumour volume when a purpose built track ball is available and the tumour morphology is relatively simple. Volume measurement should be obtained using the Cavalieri method to ensure that the estimates are unbiased and that their precision can be predicted. The measured tumour regression rates may have important implications for improving local tumour control, optimum timing of brachytherapy and minimizing the risk of radiation damage.

Adult↗
PubMed2005

Pantak Therapax SXT 150: performance assessment and dose determination using IAEA TRS-398 protocol.

The performance assessment and beam characteristics of the Therapax SXT 150 unit, which encompass both low and medium-energy beams, were evaluated. Dose determination was carried out by implementing the International Atomic Energy Agency (IAEA) TRS-398 protocol and measuring all the dosimetric parameters in order to have a solid, consistent and reliable data set for the unit. Mechanical movements, interlocks and applicator characteristics agreed with specifications. The timer exhibited good accuracy and linearity. The output was very stable, with good repeatability, long-term reproducibility and no dependence on tube head orientation. The measured dosimetric parameters included beam first and second half-value layers (HVLs), absorbed dose rate to water under reference conditions, central axis depth dose distributions, output factors and beam profiles. Measured first HVLs agreed with comparable published data, but the homogeneity coefficients were low in comparison with typical values found in the literature. The timer error was significant for all filters and should be taken into consideration for the absorbed dose rate determination under reference conditions as well as for the calculation of treatment times. Percentage depth-dose (PDD) measurements are strongly recommended for each filter-applicator combination. The output factor definition of the IAEA TRS-398 protocol for medium-energy X-ray qualities involves the use of data that is difficult to measure. Beam profiles had small penumbras and good symmetry and flatness except for the lowest energy beam, for which a heel effect was observed.

Calibration↗
PubMed2002

The contour of the femoral head-neck junction as a predictor for the risk of anterior impingement.

Impingement by prominence at the femoral head-neck junction on the anterior acetabular rim may cause early osteoarthritis. Our aim was to develop a simple method to describe concavity at this junction, and then to test it by its ability to distinguish quantitatively a group of patients with clinical evidence of impingement from asymptomatic individuals who had normal hips on examination. MR scans of 39 patients with groin pain, decreased internal rotation and a positive impingement test were compared with those of 35 asymptomatic control subjects. The waist of the femoral head-neck junction was identified on tilted axial MR scans passing through the centre of the head. The anterior margin of the waist of the femoral neck was defined and measured by an angle (alpha). In addition, the width of the femoral head-neck junction was measured at two sites. Repeated measurements showed good reproducibility among four observers. The angle alpha averaged 74.0 degrees for the patients and 42.0 degrees for the control group (p < 0.001). Significant differences were also found between the patient and control groups for the scaled width of the femoral neck at both sites. Using standardised MRI, the symptomatic hips of patients who have impingement have significantly less concavity at the femoral head-neck junction than do normal hips. This test may be of value in patients with loss of internal rotation for which a cause is not found.

Adolescent↗
PubMed1989

On the performance and reliability of mechanized urine teststrip measurement in comparison with visual reading.

Prescreening of urine specimens by teststrips is a valuable procedure for reducing the work load of the urine analysis laboratory: positive results for leukocytes, erythrocytes (haemoglobin), protein, and/or nitrite are widely used to select pathological specimens for subsequent microscopic examination. By standardization of the measurement conditions, mechanized teststrip reading is claimed to give more reproducible results than conventional techniques. To assess their ability to improve urine prescreening, especially with regard to the comparability of the results, the practical and analytical performance of three commercially available analysers (Rapimat II/T from Behringwerke AG, Urotron RL9 from Boehringer Mannheim GmbH, and Clinitek 200 from Ames/Bayer Diagnostic) was compared with visual reading. Analytical criteria were assessed using routine urine samples, while reproducibility was tested by repeated analysis of three different commercial control urines (Kova Trol from Madaus). A mean imprecision between 3% and 11.9% was found for the mechanized dipstick reading which was comparable to that found with visual examination (4.5% with Combur9-teststrips, Boehringer Mannheim GmbH). Due to the crude classification of the results, the different analysers as well as the visual technique gave quite different distributions for each of the semiquantitative parameters in the same urine samples. Even if statistical analysis was restricted to the frequency of positive results only, significant differences (chi 2-test, p less than 0.001-0.05) between methods were obtained, but these differences could not be attributed to one method alone. Using microscopic sediment analysis as reference, pathological urines were detected with a comparable sensitivity/specificity: Urotron 0.85/0.84, Rapimat 0.91/0.67, Clinitek 0.82/0.81, and duplicate visual reading 0.88/0.67 and 0.91/0.93. Mechanized teststrip reading had no obvious advantage with respect to the time required. We conclude: (i) no improvement in analytical performance or in speed of analysis could be claimed for mechanized methods in comparison with visual reading; (ii) mechanized teststrip reading might decrease the work load of the urine laboratory if integrated into a computerized laboratory system; (iii) mechanized teststrip reading will become analytically advantageous over visual reading if a more refined classification of the results is achieved.

Evaluation Studies as Topic↗
PubMed1996

In praise of the hyperglycemic clamp. A method for assessment of beta-cell sensitivity and insulin resistance.

The most widely used methods for the assessment of beta-cell response and peripheral tissue sensitivity to insulin include the oral glucose tolerance test (OGTT), the frequently sampled intravenous glucose tolerance test, and the hyperinsulinemiceuglycemic clamp technique. During an OGTT, glucose levels increase after a variable lag period, then reach a peak and fall variably among individuals. The response even varies in the same subject upon repeat testing. A more reproducible glucose curve is achieved with an intravenous glucose tolerance test in which the plasma glucose levels rise rapidly to a very high level and fall exponentially. In neither of the two methods is a steady-state glucose level achieved. In the hyperinsulinemic-euglycemic clamp technique, a steady-state glucose level can be maintained at any level of hyperinsulinemia. However, an assessment of beta-cell sensitivity is not obtained. The less used hyperglycemic clamp technique can assess beta-cell sensitivity as well as peripheral tissue sensitivity. Moreover, a measure of glucose effectiveness or non-insulin-mediated glucose uptake can also be determined. With this technique the beta-cells of all subjects are stimulated with the same arterial glucose concentration, thus enabling assessment of beta-cell response to identical plasma glucose levels. Comparison of responses to stable hyperglycemic stimuli can be made in glucose-tolerant and -intolerant states with the addition of various substances, either alone or in combination. The use of the hyperglycemic clamp and several of its variant forms is reviewed as an alternative method for assessment of glucose homeostasis.

Adult↗
PubMed2000

Effects of learning and movement frequency on polyrhythmic tapping performance.

This study examined the effect of learning a complex bimanual coordination task at different movement frequencies. 30 subjects performed 5:3 polyrhythmic tapping at either high, medium, or low movement frequency on a rhythmic synchronization task and then reproduced the polyrhythmic pattern repeatedly in the spontaneous task. Analysis showed that practice on the synchronization task qualitatively changed correct responses into anticipatory ones. The synchronization learning of the polyrhythm caused the anticipatory responses and so, may involve memorization of serial positions within the polyrhythm. Also, more anticipatory responses were indicated in performance at the medium and low frequencies than at the high frequency on the synchronization task. In addition, deviations of taps from expected tapping positions were observed in performance of the spontaneous task at the high frequency. These results suggest that the movement frequency qualitatively influenced the learning of this bimanual coordination. Especially at the high frequency, frequent shifts to other coordination patterns occurred on the spontaneous task. This means that the performance at higher frequency is more strongly affected by entrainment between the two hands.

Adolescent↗
PubMed1983

Evoked acoustic emissions from the human ear. III. Findings in neonates.

Stimulated acoustic emissions were recorded in a consecutive series of 20 full-term and otherwise normal neonates with the equipment and method previously used in adults. One ear randomly chosen was tested in each baby, and otoscopy and tympanometry were normal in all ears. A 2 kHz click stimulus was presented with a repetition rate of 10/sec and the recordings were performed at three intensities, i.e. 70, 50 and -20 dBatt (dBatt approximately dB p.e. SPL). The 50 dB recording was repeated for check of reproducibility. A clear and reproducible response could be identified from all ears at 50 dBatt. However, as in the adults, the response pattern differed significantly from one ear to another, both regarding the number of 'echoes', their latencies, response amplitudes, and frequency content. The 'echo' group latencies and amplitudes were within the same range as in normal adults and the amplitude input-output curves exhibited a clear non-linearity. The relationship between latency and frequency was just as ambiguous as in the adults. Also, in the neonates, the cross correlation analysis proved to be an efficient method to indicate whether or not a true response was present. The results from this investigation are compared with those described in the literature from other audiological tests and it is concluded that the recording of the stimulated acoustic emissions could be applicable as a screening procedure in newborns.

Acoustic Stimulation↗
PubMed1993

Comparative study of transcranial color duplex sonography and transcranial Doppler sonography in adults.

To determine whether the frequency shift recorded in basal cerebral arteries corresponds to "true" flow velocities, a prospective comparative study of transcranial color duplex sonography (TCCD) and transcranial Doppler sonography (TCD) was performed. A 2.0-MHz transducer of a computerized TCCD system and a TCD device were used. The middle cerebral artery (MCA) and anterior cerebral artery (ACA) were examined by TCCD in 49 healthy volunteers (mean age 35 +/- 12 years). In 45 of the same volunteers a comparative TCD examination was possible. The studies were carried out blindly by different examiners at separate appointments. Peak systolic flow velocity, end-diastolic maximum flow velocity, time-averaged maximum flow velocity, and the pulsatility index were measured by both techniques. Additionally, for TCCD, time-averaged flow velocity was assessed, the resistance index and a spectral broadening index were calculated, and the energy output required for reliable measurement was analyzed. The TCCD signals were recorded in 98% of both MCA's and ACA's; with TCD, signals were recorded in 98% of MCA's and 87% of ACA's. Although in both vessels the angle-corrected peak systolic and time-averaged maximum velocities were approximately 10% to 15% higher in TCCD than in TCD measurements, correlation of flow velocities between both techniques was significant (p < 0.0001); differences between sides and age-dependency of flow velocities corresponded as well. In a reproducibility study, TCCD was repeated in 27 subjects by a third examiner with significant correlation (p < 0.0001) of both TCCD examinations. It is concluded that the advantage of TCCD is associated more with a qualitative aspect than a quantitative one. The additional visual dimension of TCCD can open new diagnostic possibilities in cerebrovascular disorders.

Adult↗