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PubMed1978

A new method of multiplanar emission tomography using a seven pinhole collimator and an Anger scintillation camera.

A new method of multiplanar emission tomography is described; it uses a wide-field Anger scintillation camera (37.5 cm crystal diameter) and a seven-pinhole collimator. The pinholes (5.5 mm) acquire data simultaneously from the emissing source and project the data onto seven independent regions of the camera crystal. Multiple planes are reconstructed from the initial seven-view data acquisition through the use of a computerized addition-multiplication algorithm and variation of the superposition relationships among the projected views. These planes are then altered iteratively by a least-error criterion following ray-sum comparison with the original views. Planar resolution (full-width-half-maximum) is 1.0 cm and depth resolution is 1.5 cm. In 42 patients with angiographically demonstrated coronary-artery disease, studies of myocardial Tl-201 perfusion, under exercise, have shown improved detection sensitivity in comparison with scintigraphy using parallel-hole collimation.

Coronary Disease↗
PubMed1992

Relationship between cytokine-dependent cell cycle progression and MHC class II antigen expression by human CD34+ HLA-DR- bone marrow cells.

Human CD34+ HLA-DR- bone marrow cells constitute a phenotypically homogeneous population of quiescent cells. More than 97% of CD34+ HLA-DR- cells reside in the G0/G1 phase of the cell cycle. The in vitro effects of two cytokines, IL-1 alpha and IL-3, alone or in combination, on the viability, cell cycle status and acquisition of HLA-DR by this cell population were examined. Cell viability was preserved in cultures receiving cytokines, but declined steadily in cultures deprived of exogenous IL. Over a period of 4 days, IL-3 progressively induced the expression of HLA-DR although driving corresponding numbers of cells into S and G2 + M. Although IL-1 alpha induced the expression of HLA-DR, it was not as effective as IL-3 in promoting the exit of these cells from G0/G1. Combinations of IL-1 alpha and IL-3, however, exerted an even greater effect on promoting both HLA-DR expression and entry of cells into active phases of the cell cycle. Simultaneous measurement of HLA-DR expression and cell cycle status in response to IL-1 alpha and IL-3 indicated that the majority of de novo expression of HLA-DR occurred in cells that remained in G0/G1. CD34+ HLA-DR- cells cultured with IL-1 alpha and IL-3 but arrested in G0/G1 by hydroxyurea were still capable of expressing HLA-DR, demonstrating that the acquisition of HLA-DR was independent of the entry of these cells into active phases of the cell cycle. These data indicate that the survival, HLA-DR expression, and cell cycle status of human CD34+ HLA-DR- bone marrow cells are governed by regulatory cytokines such as IL-1 alpha and IL-3. In addition, the entry of these cells into active phases of the cell cycle does not seem to be a prerequisite for the expression of HLA-DR, nor does it seem that the acquisition of HLA-DR by hematopoietic progenitor cells is a marker of cells entering the S phase of the cell cycle.

Antigens, CD↗
PubMedDate not supplied

The incidence of Helicobacter pylori infection is not increased among obese young individuals in Greece.

GOALS: To identify possible risk factors affecting the acquisition of Helicobacter pylori (Hp) infection and to investigate whether the incidence of infection is higher among obese and overweight verus normal-weight young adults in Greece. STUDY: Serum was obtained from 224 young male Navy recruits (mean age, 22.84 years) during their induction into the Hellenic Navy. An enzyme-linked immunosorbent assay to detect Hp-specific IgG serum antibodies, as well as gastroscopy with biopsy, were used to identify the infected individuals. A structured questionnaire was filled out for each subject regarding environmental conditions, socioeconomic conditions, dietary habits, and data related to their personal and family health history. RESULTS: H. pylori positivity rate was 27.23%. Univariate analysis recognized that the number of siblings in the same bedroom was significantly higher among Hp-positive than Hp-negative individuals. Logistic regression analysis showed that sharing the same bedroom with more than one sibling during childhood and consumption of fast food are independent predictors of Hp acquisition. The presence of obesity (body mass index > or =25 kg/m2) remained unrelated to the Hp status of the individuals. CONCLUSIONS: These data suggest that the risk of Hp infection does not increase in overweight young persons. Sharing the same bedroom with more than one sibling during childhood is an important determinant in acquiring Hp infection. Increased fast food consumption could be an important source of the infection outside of the home.

Adult↗
PubMed2003

Passing on tacit knowledge in anaesthesia: a qualitative study.

OBJECTIVE: To explore the acquisition of knowledge in anaesthetic practice using qualitative methods. METHODS: An ethnographic study examining the nature of expertise in anaesthesia in one English hospital. The study used qualitative research methods, including observation of anaesthetic practice and interviews with members of the anaesthetic team. An integral part of the study was a process of feedback to the anaesthetic team including presenting observational data and conducting debriefing interviews with individual team members. RESULTS: The study highlights the continued importance of the clinical apprenticeship in passing on knowledge, but also emphasizes the central role of practising independently in the acquisition of skills. Anaesthetists who participated in debriefing interviews or read observational transcripts found the experience valuable for thinking about their own practice. DISCUSSION: One suggestion arising from the use of qualitative methods in this setting is that the type of detailed, systematic observation and data recording used in this study could be beneficial in the training and, possibly, appraisal of anaesthetists. This novel and innovative application of qualitative methods in anaesthesia is described and discussed with a view to broadening the debate about specialist training.

Anesthesia↗
PubMed2000

Trainable rule-based algorithm for the measurement of joint space width in digital radiographic images of the knee.

The progression of osteoarthritis (OA) can be monitored by measuring the minimum joint space width (mJSW) between the edges of the femoral condyle and the tibial plateau on radiographs of the knee. This is generally performed by a trained physician using a graduated magnifying lens and is prone to the subjectivity and variation associated with observer measurement. We have developed software that performs this measurement automatically on digitized radiographs. The test data consisted of 180 digitized radiographs of the knee (90 duplicate acquisitions) from 18 normal (nonarthritic) subjects and 38 images from 10 subjects with OA. These were digitized and manually cropped so that the images were free of nonanatomical structures and the knee was approximately centered. The software first determined the edge of the femoral condyle on 400 microm pixel subsampled images. Contours marking the location of the tibial plateau in the medial compartment were found on 100 microm images using the femoral edge as a reference. The algorithm was trained using an independent but similar data set and using a jackknife approach with the test data. The results were compared to contours drawn by a trained reader and the duplicate acquisitions were used to measure the reproducibility of the mJSW measurement. The reproducibility was 0.16 mm and 0.18 mm for normal and osteoarthritic knees, respectively, representing an improvement of approximately a factor of 2 over manual measurement. The algorithm also showed excellent agreement with the hand-drawn contours and with mJSW determined by the manual method.

Algorithms↗
PubMed2000

Measurement of volumetric mitral and aortic blood flow based on a new freehand three-dimensional colour flow imaging method. An in vivo validation.

AIMS: To validate a new three-dimensional (3D) colour flow method used to calculate cardiac output (CO) in aortic and mitral blood flow. METHODS: The transducer was freely tilted transthoracically using a magnetic locating device recording its spatial position. Raw digital ultrasound data were recorded in healthy subjects during 10-20 heartbeats at a high frame rate ranging from 41 to 66 frames/s and analysed off-line with no loss in temporal resolution. Blood flow velocities aligned with the ultrasound beam were integrated across a moving spherical surface to calculate volumetric flow. RESULTS: The range of agreement between the 3D mitral and 3D aortic method was 0.04+/-1.32 l/min (mean+/-2 standard deviations). The range of agreement between 3D aortic flow and the two-dimensional (2D) pulsed wave Doppler method (2DPW) in the left ventricular outflow tract (LVOT) was 0.7+/-1.7 l/min, while the range of agreement between 3D mitral flow and the 2DPW method was 0.88+/-1.64 l/min. CONCLUSION: The 3D methods agreed well. The 3D volumetric flow overestimated the 2DPW method, as expected, and the range of agreement was wide. The common pitfalls in pulsed wave ultrasound methods to calculate CO were avoided, as the 3D method was angle-independent, no assumptions about the velocity profile were made, and a moving sample surface was applied. The acquisition of data was fast and easy and high temporal resolution was achieved.

Adult↗
PubMed2006

Description and implementation of a quality control program in an imaging-based clinical trial.

RATIONALE AND OBJECTIVES: The American College of Radiology Imaging Network is participating in the National Lung Screening Trial, a large, multicenter, randomized controlled trial, comparing multidetector helical computed tomography (MDCT) versus chest radiography (CXR) in screening for lung cancer. Because the threshold for detection of disease is an inherent function of image quality, and consistent image quality is necessary to track changes in suspicious findings, our purpose was to develop an image quality control (QC) program across all clinical sites for both modalities. MATERIALS AND METHODS: The primary goals of the QC program include standardization of imaging protocols, certification of imaging equipment, and ongoing, periodic evaluation of the equipment calibration and image quality. Minimum standards for equipment and standardized cross-platform acquisition protocols are achieved via radiologist and physicist attestation forms and web-distributed technique charts, respectively. Imaging equipment performance standards are implemented through an initial machine certification process that includes equipment calibration. Ongoing assessment of equipment performance and calibration, as well as adherence to established imaging protocols. is accomplished via periodic submission of calibration records and phantom images. Participant-specific image acquisition parameters are entered into a web-based centralized database and variations from established protocols are automatically flagged for review. Participant radiation dose can be estimated from the image acquisition parameters applied to the imaging equipment calibration measurements. A radiologist visual review committee also evaluates participant images for diagnostic quality. Data are collected from 23 independent centers, representing 14 models of MDCT scanners from four manufacturers, and CXR systems that include film-screen, computed radiography, and direct digital radiography systems. RESULTS: Widespread imaging protocol variation in extant clinical practice-as well as variability in equipment technology, image acquisition parameters, manufacturer terminology, and user interface-have required careful standardization as a prerequisite to trial participation and ongoing image QC. Acceptable ranges for image acquisition parameters have been refined to accommodate continuously evolving equipment platforms and the scope of participant size and body habitus. CONCLUSION: Standardization of imaging protocols is a critical component of image-based clinical trials, predicated on ongoing dialogue between sites and a centralized review committee.

Artificial Intelligence↗
PubMed2004

Latest techniques in head and neck CT angiography.

Continuous evolution of multi row CT is increasingly making CT angiography a viable imaging modality for assessment of the supraaortic and intracranial vessels as an anatomically and functionally coherent vascular system. Extended non-invasive examinations with reduced contrast volume have become feasible with the availability of 16 and 64 row MDCT scanners. Prerequisites to obtain high resolution CT angiographies of the head and neck vessels with superior detail include the administration of low contrast volume, high contrast density (400 mg I/ml) contrast media, adequate timing and data acquisition, optimal flow rate (4 ml/s) and saline flushing. Non-invasiveness, delineation of vessel calcification, virtual independence from hemodynamic conditions, and the ability to provide quantification without needing to correct for magnification are all attributes that favour CT angiography over digital subtraction angiography and to some extent even magnetic resonance angiography as an alternative non-invasive technique. CT angiography is established as a modality of choice for the assessment of patients with acute stroke and chronic steno-occlusive disease. CT angiography may indicate the presence of extra- or intracranial acute vessel occlusion and dissection, predisposing atherosclerotic steno-occlusive disease and thus indicate thrombo-embolism or local appositional thrombosis as the principle pathogenic factor. CT angiography is used to assess anatomy, and to depict the presence, location and extent of calcified and non-calcified plaque as a cause of high grade stenosis. Despite relatively limited sensitivity CT angiography is indicated for suspected or confirmed aneurysms that demand further verification of their presence, geometry, or relationship to parent artery branches and osseous anatomic landmarks. Low volume high density contrast media have substantially increased the ability of CT angiography to depict small aneurysms, small branches, and collateral vessels, and to recognize the residual lumen in vessels with high grade stenosis or conditions such as dissection or pseudo-occlusion. Superior detail high resolution CT angiography is thus a viable alternative to DSA, relegating the latter technique to endovascular treatment applications only.

Angiography↗
PubMed2004

Acquisition of multidrug-resistant Pseudomonas aeruginosa in patients in intensive care units: role of antibiotics with antipseudomonal activity.

A matched case-control study was performed to identify risk factors for acquiring multidrug-resistant Pseudomonas aeruginosa (MDRPA) in intensive care unit (ICU) patients during a 2-year period. MDRPA was defined as P. aeruginosa with combined decreased susceptibility to piperacillin, ceftazidime, imipenem, and ciprofloxacin. Thirty-seven patients who were colonized or infected with MDRPA were identified, 34 of whom were matched with 34 control patients who had cultures that showed no growth of P. aeruginosa. Matching criteria were severity of illness and length of ICU stay, with each control patient staying in the ICU for at least as long as the time period between the corresponding case patient's admission to the ICU and the acquisition of MDRPA. Baseline demographic and clinical characteristics and the use of invasive procedures were similar for case patients and control patients. Multivariate analysis identified duration of ciprofloxacin treatment as an independent risk factor for MDRPA acquisition, whereas the duration of treatment with imipenem was of borderline significance. These data support a major role for the use of antibiotics with high antipseudomonal activity, particularly ciprofloxacin, in the emergence of MDRPA.

Aged↗
PubMed1999

Taking MAD to the extreme: ultrafast protein structure determination.

Multiwavelength anomalous diffraction data were measured in 23 min from a 16 kDa selenomethionyl substituted protein, producing experimental phases to 2.25 A resolution. The data were collected on a mosaic 3 x 3 charge-coupled device using undulator radiation from the Structural Biology Center 19ID beamline at the Argonne National Laboratory's Advanced Photon Source. The phases were independently obtained semiautomatically by two crystallographic program suites, CCP4 and CNS. The quality and speed of this data acquisition exemplify the opportunities at third-generation synchrotron sources for high-throughput protein crystal structure determination.

Base Sequence↗
PubMed2004

The reproducible acquisition of comparative liquid chromatography/tandem mass spectrometry data from complex biological samples.

An in-depth study of the reproducibility of data acquired for comparative proteomics analysis using a prototype two-stage heated laminar flow chamber fitted to a commercial high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) instrument was undertaken. The study is based on 24 replicate samples from four independent membrane preparations derived from two matched breast cancer cell lines. Variation and reproducibility in the data were evaluated at several levels highlighting the relative efficiency and variability of the acquisition routines used. Specifically, variation in the number and relative intensities of chromatographic peaks eluted from the LC column, precursor ion selection and sequence identification were evaluated. On average, approximately 6500 chromatographic peaks were generated for each acquisition with a corresponding coefficient of variance (CV) of less than 20%. Precursor ion selection and sequence identification averaged 1380 and 780 events per acquisition sample, respectively, with corresponding CVs of less than 10% for each. The reproducibility in the precursor ion selection was typically better than 60% between similar replicates. Using protein and peptide internal standards, it was found that the CV in retention time across the gradient between two acquisition pairs was typically less than 5%, whereas the average intensity ratio was 1.0 (expected) with a CV approaching 20%. An evaluation of the intensity ratios calculated from endogenous peptide sequences, identified across the acquisition set, indicated a CV of approximately 30%. Similarly, the CV associated with the top 1000 peptides indicated a mean and median of 28.4 and 26.95%. For a given acquisition pair it was also found that approximately 11% of the chromatographic peaks eluting from the column were linked to a sequence or identified. For these experiments, less than 10% of the peak pairs had absolute ratios greater than 2.0 and of those only approximately 10% had sequences linked to them. For each matched acquisition set on average 406 proteins were identified with a CV of less than 10%. Of the proteins that were identified approximately 30% had at least one predicted trans-membrane domain, indicating a four-fold increase over a crude homogenate sample with only minor enrichment. During these experiments it was found that the interface did not significantly alter the relative charge state distribution of ions, nor did it introduce significant interference from background ions. The interface was capable of 24-hour acquisition cycles.

Amino Acid Sequence↗
PubMed2003

Male circumcision for prevention of heterosexual acquisition of HIV in men.

BACKGROUND: The findings from observational studies, reviews and meta-analyses, supported by biological theories, that circumcised men appear less likely to acquire human immunodeficiency virus (HIV) has contributed to the recent ground swell of support for considering male circumcision as a strategy for preventing sexually acquired infection. We sought to elucidate and appraise the global evidence from published and unpublished studies that circumcision can be used as an intervention to prevent HIV infection. OBJECTIVES: 1) To assess the evidence of an interventional effect of male circumcision for preventing acquisition of HIV-1 and HIV-2 by men through heterosexual intercourse 2) To examine the feasibility and value of performing individual person data (IPD) meta-analysis SEARCH STRATEGY: We searched online for published and unpublished studies in The Cochrane Library (issue 2, 2002), MEDLINE (April 2002), EMBASE (February 2002) and AIDSLINE (August 2001). We also searched databases listing conference abstracts, scanned reference lists of articles and contacted authors of included studies. SELECTION CRITERIA: We searched for randomized and quasi-randomized controlled trials of male circumcision or, in their absence, observational studies that compare acquisition rates of HIV-1 and HIV-2 infection in circumcised and uncircumcised heterosexual men. DATA COLLECTION AND ANALYSIS: Independent reviewers selected studies, assessed study quality and extracted data. We stratified studies based on study design and on whether they included participants from the general population or high-risk groups (such as patients treated for sexually transmitted infections). We expressed findings as crude and adjusted odds ratios (OR) together with their 95% confidence intervals (CI) and conducted a sensitivity analysis to explore the effect of adjustment on study results. We investigated whether the method of circumcision ascertainment influenced study outcomes. MAIN RESULTS: We identified no completed randomized controlled trials. Three randomized controlled trials are currently underway or commencing shortly. We found 34 observational studies: 16 conducted in the general population and 18 in high-risk populations. It seems unlikely that potential confounding factors were completely accounted for in any of the included studies. In particular, important risk factors, such as religion and sexual practices, were not adequately accounted for in many of the included studies. General population study results:The single cohort study (N = 5516) showed a significant difference in HIV transmission rates between circumcised and uncircumcised men [OR = 0.58; 95% CI: 0.36 to 0.96]. Results for the 14 cross-sectional studies were inconsistent, with point estimates for unadjusted odds ratios varying between 0.28 and 1.73. Six studies had statistically significant results, four in the direction of benefit and two in the direction of harm. The test for heterogeneity between the cross-sectional studies was highly significant (chi-square = 77.59; df = 13; P-value < 0.00001). Nine studies reported adjusted odds ratios with eight in the direction of benefit, ranging from 0.26 to 0.80. Use of adjusted results tended to show stronger evidence of an association although they remained heterogenous (chi-square = 75.2; df = 13; P-value < 0.00001). Only one case-control study was found (N = 51) which had a non-significant result [OR = 1.90; 95% CI: 0.50 to 7.20]. High-risk group study results:The four cohort studies identified found a protective effect from circumcision with point estimates for unadjusted odds ratios varying from 0.10 to 0.39. Two of these studies had statistically significant results. Two studies reported adjusted odds ratios, both protective with one being significant. The chi-square test for between-study heterogeneity was not significant (chi-square = 5.21; df = 3; P-value = 0.16). All eleven cross-sectional studies reporting unadjusted results found benefit from circumcision, eight of which had statistically significant results. Estimates of effnal studies reporting unadjusted results found benefit from circumcision, eight of which had statistically significant results. Estimates of effect varied from an unadjusted odds ratio of 0.10 to 0.66. Between-study heterogeneity was significant with the chi-square = 29.77; df = 10; P-value = 0.0009. Four of these studies reported adjusted odds ratios ranging from 0.20 to 0.59 and all were significant. One additional cross-sectional study only reported an adjusted odds ratio in the direction of benefit which was statistically significant. All three case-control studies found a protective effect of circumcision on HIV status, two being statistically significant. Point estimates varied from unadjusted odds ratios of 0.37 to 0.88. One reported an adjusted odds ratio showing a significant protective effect. Adverse effects:No studies reported on the adverse effects of circumcision. In most studies, circumcision had taken place during childhood or adolescence before the studies commenced. REVIEWER'S CONCLUSIONS: We found insufficient evidence to support an interventional effect of male circumcision on HIV acquisition in heterosexual men. The results from existing observational studies show a strong epidemiological association between male circumcision and prevention of HIV, especially among high-risk groups. However, observational studies are inherently limited by confounding which is unlikely to be fully adjusted for. In the light of forthcoming results from RCTs, the value of IPD analysis of the included studies is doubtful. The results of these trials will need to be carefully considered before circumcision is implemented as a public health intervention for prevention of sexually transmitted HIV.

Circumcision, Male↗
PubMedDate not supplied

Semi-automatic medical image processing.

During the last few years a number of systems for graphics generation and image processing have been developed by the authors. Depending on the application, one can consider many kinds of systems; for example for simple analysis of cellular images, a system that realizes the video mixing between a camera and graphics data from a microcomputer would be adequate. For a more elaborate analysis, such as angiographic and echographic images requiring a high level of interactivity, a semi-automatic acquisition system linked to a host computer would be necessary. Limits in precision and execution time of these semi-automatic systems leads to independent work-stations that realize digital acquisition, processing, and display of images.

Computers↗
PubMedDate not supplied

Molecular identification of algal endosymbionts in large miliolid Foraminifera: 2. Dinofiagellates.

Large miliolid foraminifers of the subfamily Soritinae bear symbiotic dinoflagellates morphologically similar to the species of the "Symbiodinium" complex, commonly found in corals and other marine invertebrates. Soritid foraminifers are abundant in coral reefs and it has been proposed that they share their symbionts with other dinoflagellate-bearing reef dwellers. In order to test this hypothesis, we have analysed partial large subunit ribosomal DNA sequences from dinoflagellates symbionts obtained from 28 foraminiferal specimens, and compared them to the corresponding sequences of Symbiodinium-like endosymbionts from various groups of invertebrates. Phylogenetic analysis of our data shows that all soritid symbionts belong to the "Symbiodinium" species complex, within which they form seven different molecular types (Frl-Fr7). Only one of these types (Fr1) branches within a group of invertebrate symbionts, previously described as type C. The remaining six types form sister groups to coral symbionts previously designed as types B, C, and D. Our data indicate a high genetic diversity and specificity of Symbiodinium-like symbionts in soritids. Except for type C, we have found no evidence for the transmission of symbionts between foraminifers and other symbiont-bearing invertebrates from the same localities. However, exchanges must have occurred frequently between the different species of Soritinae, as suggested by the lack of host specificity and some biogeographical patterns observed in symbiont distribution. Our data suggest that members of the subfamily Soritinae acquired their symbionts at least three times during their history, each acquisition being followed by a rapid diversification and independent radiation of symbionts within the foraminiferal hosts.

Animals↗
PubMedDate not supplied

Three-dimensional, isotropic MRI: a unified approach to quantification and visualization in congenital heart disease.

PURPOSE: Current standards in magnetic resonance imaging of congenital heart disease are based mostly on anisotropic protocols to image both morphology and function. Operator-dependent acquisition planning is typically needed to obtain the desired images. We propose to instead use operator-independent, three-dimensional, isotropic imaging protocols to acquire both morphology and function (cine and flow) of the entire heart in a few standardized acquisitions. Subsequently, due to the isotropic property of the data, any desired imaging plane can be "imaged" offline by interactive planar reformatting and used for qualitative and quantitative diagnostic evaluation. MATERIALS AND METHODS: Morphological data was acquired in patients using 3D steady state free precession (SSFP) protocols, and functional data in volunteers using multislice 2D or 3D cine SSFP as well as 3D, three-component phase-contrast velocity mapping with EPI readouts. Tools to integrate morphological and functional offline image evaluation based on interactive planar reformatting, volume rendering, and corresponding quantification tools were implemented and discussed. RESULTS: We successfully acquired and integrated morphology and flow and demonstrated potential clinical applications. CONCLUSION: User independent acquisitions of morphological and functional isotropic 3D datasets with real-time, interactive planar reformatting, volume rendering, and integration of morphology and function, have the potential to replace conventional, user dependent, anisotropic 2D imaging in patients with cardiac malformations.

Adolescent↗
PubMed1991

Computerized acquisition and evaluation of whole blood aggregometry data.

A special-purpose software package has been developed to acquire and analyze analog signals from a whole blood aggregometer. Up to four independent blood samples can be analyzed simultaneously or with arbitrary delays in time. The user menu ensures proper calibration prior to each measurement and provides a real-time graphic display of the signals. From analog/digital converted data, several "derived diagnostic quantities" are calculated. They are constructed with the aim to mimic the process of visual inspection of aggregometry curves and thus place the intuitive diagnostics of blood platelet function on a quantitative basis.

Computer Graphics↗
PubMed2002

Flow cytometric CD34+ determination in stem cell transplantation: before or after cryopreservation of grafts?

Various attempts have been made to standardize and improve the reproducibility of flow cytometric determination of CD34+ hematopoietic progenitor cells. It is still not clear, however, whether the quantification of CD34+ cells in a stem cell graft should be done before or after cryopreservation. To address this issue, we investigated 78 unselected and 32 immunomagnetically selected autologous and allogeneic leukapheresis products (LA) before and after cryopreservation using pilot vials. Cell numbers were quantified within a Neubauer chamber, and CD34+ content was determined by flow cytometry; propidium iodide staining was used to exclude dead cells from analysis. Before freezing, the mean viable CD34 cell content in the unselected samples was 1.22% and increased after thawing to a mean of 2.16% of viable cells. Taking into account cell loss and cell death, the overall recovery of viable cells was 64.5%; all CD34+ cells could be recovered. Mean purity in the CD34-selected cell fraction was 85% (48-97) before and 91.3% (67-99) after thawing. The number of viable cells was 86.8% before and 86.1% after freezing with a 93.9% recovery of total cells. This leads to a mean 93.7% (SD +/- 23.1) recovery of viable cells and 100% (SD +/- 22.3) recovery of viable CD34+ cells. There was no significant difference in tolerance to freeze/thaw stress between cells from heavily pretreated autologous patients and healthy allogeneic donors. Our data show that freezing significantly increases the percentage of CD34(+) cells in unmanipulated LA, probably due to the death of granulocytes and mononuclear cells (MNCs). Nevertheless, the overall number of viable CD34+ cells in unselected as well as selected samples remains unchanged. Thus, CD34 data from different laboratories, for example, within multicenter trials, should be comparable independent of the different time points of acquisition.

Antigens, CD34↗
PubMed1988

Maturational influences on perseveration of avoidance and reversal learning after selected brain damage in rats.

In Experiment I, 18 weanling and 18 adult male rats received bilateral electrolytic lesions of the dorsomedial or ventrolateral prefrontal cortex, or the hippocampus, while 12 additional pups and adults served as nonoperated controls (n = 6/group). Subjects were observed for perseverative responding in acquisition of a stepup avoidance task, followed by reversal training and extinction. Thirty days later, subjects were retrained and tested in the same manner. During initial training, the numbers of trials to criteria, errors, and latencies on the last 5 trials all indicated significant effects from age, primarily, and surgery, secondarily. After 30 days, surgical effects assumed a more dominant role, with hippocampal and ventrolateral damage producing the greatest extent of response perseveration. Experiment II replicated the essential procedural sequence as Experiment I, but included damage from combined lesions. 24 weanlings and 24 adults received bilateral lesions to the dorsomedial prefrontal cortex, the dorsomedial prefrontal cortex plus caudate nucleus, or sham lesions. Subjects from this experiment were trained after 7 days or 60 days recovery (n = 6/group). The acquisition results indicated that all independent variables were significant, but only the age effect attained significance in the extinction data. Both experiments point to the profound influence of age at the time of damage in accounting for recovery of avoidance behavior.

Aging↗