Biomedical subjects
Publications and source records attributed to G Schulz.
The synthesis of eight rigid analogs of L-N-iminoethylornithine (L-NIO) is described. The compounds have been evaluated for their inhibition of inducible nitric oxide synthase. Preliminary structure-activity relationships are discussed.
Defined serum-free conditions have great conceptual advantages for the biological safety and standardization of clinical gene transfer into hematopoietic stem cells. In the only study reported to date, Sekhar et al. achieved low serum conditions by a complex concentration procedure of a retroviral supernatant initially containing 10% fetal bovine serum. The high cost, small volume, possible coenrichment of serum-derived pathogens, limited recovery of vector particles, and low titer of the final diluted medium restrict the clinical application of this procedure. Transduction of primitive hematopoietic progenitor cells was not demonstrated. In the present study, a defined serum-free medium containing high titers of the pseudotyped retroviral vector PG13/LN was generated from PG13/LN producer cells without requiring a physical enrichment procedure. The transduction of committed hematopoietic progenitor cells in the serum-free vector-containing medium was efficient, and similar to that occurring under serum-containing control conditions. The number of primitive human hematopoietic long-term culture-initiating cell-derived colonies (LTC-IC-derived colonies) generated from CD34+ and CD34+/HLA-DRlo peripheral blood progenitor "stem" cells (PBSCs) increased during 7 days of treatment in this vector-containing medium in the presence of IL-3, SCF, and flt-3 ligand. The described procedure allowed efficient transduction of LTC-IC-derived colonies generated from CD34+, CD34+/HLA-DRlo, and CD34+/CD38lo PBSCs. This is the first report to demonstrate an increase in primitive peripheral blood LTC-IC-derived colonies in vitro as well as their efficient transduction in a high-titer, serum-free vector-containing medium that can be produced exclusively from defined pharmaceutical-grade components, making it ideally suited for applications in clinical gene therapy.
Even though there have been numerous positron emission tomography (PET) activation studies on the perfusional and metabolic bases of language processing, little is known about the intracerebral functional network of language and cognitive processes. It was the aim of this study to investigate the cerebral interregional correlations during voluntary word association versus word repetition in healthy subjects to gain insight into the functional connectivity of associative speech processing. Due to individual variability in functional anatomy, the study protocol was designed as an averaged single-subject study. Eight healthy volunteers performed a verbal association task during fluorine-18 fluorodeoxyglucose (18F-FDG) PET scanning. Two different tasks were performed in randomized order: (a) word repetition (after auditory presentation of nouns) as a control condition, and (b) word association (after auditory presentation of nouns) as a specific semantic activation. The regional metabolic rate of glucose (rMRGlu) was calculated after brain regionalization [112 regions of interest on individual 3D flash magnetic resonance imaging (MRI)] and PET/MRI realignment. Statistical analysis was performed for comparison of association and repetition and for calculation of interregional correlation coefficients during both tasks. Compared with word repetition, word association was associated with significant increases in rMRGlu in the left prefrontal cortex, the left frontal operculum (Broca's area) and the left insula, indicating involvement of these areas in associative language processing. Decreased rMRGlu was found in the left posterior cingulum during word association. During word repetition, highly significant negative correlations were found between the left prefrontal cortex, the contralateral cortex areas and the ipsilateral posterior cingulum. These negative correlations were almost completely eliminated during the association task, suggesting a functional decoupling of the strict intercorrelation pattern.
Non-invasive methods to identify absent or persistent myocardial viability have gained increasing importance in the therapeutic management and risk stratification of patients with ischemic left ventricular dysfunction. Myocardial scintigraphy using thallium-201 and positron emission tomography with metabolic imaging of myocardial glucose metabolism, using fluorine-18 fluoro-deoxyglucose, are today the most widely used nuclear methods for the assessment of myocardial viability. Besides the prediction of reversible regional and global myocardial dysfunction following coronary revascularization, both methods have demonstrated the ability to identify patient subgroups who will benefit most with regard to cardiac prognosis and survival and those patients in whom coronary revascularization has a limited effect with regard to survival and cardiac events. This short review summarizes the clinical impact of both imaging modalities on today's diagnostic approach in patients with ischemic left ventricular dysfunction.
OBJECTIVES: We sought to define the effects of time on contractile function, morphology and functional recovery after coronary revascularization in patients with dysfunctional but viable (hibernating) myocardium. BACKGROUND: Functional recovery after coronary artery bypass graft surgery in patients with chronic myocardial hibernation is incomplete or delayed. The proposed cause is a progressive temporal degeneration of cardiomyocytes. METHODS: In 32 patients with multivessel coronary disease, regional wall motion analysis was performed in hypoperfused but metabolically active areas before and 6 months after bypass surgery. During bypass surgery, transmural biopsy samples were obtained from the center of the hypokinetic zone for light and electron microscopic analyses. The proposed duration of myocardial hibernation was retrospectively assessed. RESULTS: Patients with a subacute hibernating condition (<50 days) demonstrated a higher preoperative ejection fraction (EF, 50+/-8%), and a better preserved wall motion (WM) in the supraapical wall (-1.4+/-0.4) than did patients with intermediate-term (>50 days, EF 37+/-9%, p < 0.05; WM -2.4+/-1.5, p = 0.08) or chronic (>6 months, EF 40+/-14%, WM -2.7+/-0.9, p < 0.005) ischemia. Structural degeneration correlated with the duration of ischemia (r = 0.56, p < 0.05). Postoperative recovery of function was enhanced in patients with a short history of hibernation compared with patients with an intermediate-term or chronic condition (EF 60+/-10% vs. 40+/-10%, p < 0.001, and vs. 47+/-14%, p < 0.05). CONCLUSIONS: Hibernating myocardium exhibits time-dependent deterioration due to progressive structural degeneration with enhanced fibrosis. Early revascularization should be attempted to salvage the jeopardized tissue and improve postoperative outcome.
The possibility of maintaining, manipulating, and expanding human hematopoietic stem cells in ex vivo culture could help to provide patients with autologous and allogeneic stem cell transplants improved in purity and performance and could offer access to gene therapy of the hematopoietic system. Recent advances in the ex vivo culture of immature human hematopoietic progenitor cells and human hematopoietic stem cells have led to experimental evidence for the qualitative and quantitative maintenance and possible numerical expansion of hematopoietic stem cells in ex vivo culture, making ex vivo graft engineering a realistic possibility. This review summarizes recent developments in the field, their regulatory implications and their application in hematopoietic gene therapy.
UNLABELLED: Sustained myocardial ischemia with angina pectoris, electrocardiographic changes and subsequent non-Q-wave infarctions has been reported during percutaneous transluminal rotational atherectomy of complex coronary lesions. The purpose of this study was to evaluate the effect of rotational atherectomy on regional myocardial perfusion as assessed by serial 99mTc-sestamibi SPECT imaging with semiquantitative tracer uptake analysis. METHODS: Twenty-nine consecutive patients with anginal symptoms, complex coronary lesions (all Type B and Type C) and preserved left ventricular function were studied using resting 99mTc-sestamibi SPECT before rotational atherectomy, during and 2 days after the procedure. For semiquantitative computerized analysis, the left ventricular myocardium was divided into 24 regions, and regional perfusion was expressed as percentage of maximal sestamibi uptake. RESULTS: Visual analysis of scintigraphic images revealed transient perfusion defects corresponding to the revascularized vessel in 26 of 29 patients, whereas three patients had no transient hypoperfusion. During rotational atherectomy, perfusion decreased significantly (>2 s.d. below normal mean) in 3.1 +/- 2.4 regions (range 1-10). Perfusion in the territory of the revascularized vessel was 75% +/- 11% at baseline, decreased to 67% +/- 12% during rotational atherectomy (p < 0.001) and normalized again after rotational atherectomy to 78% +/- 8% (p < 0.001). Similarly, perfusion in the region with the maximal reduction decreased from 74% +/- 15% at baseline to 55% +/- 14% (p < 0.001) during the procedure and returned to 74% +/- 16% (p < 0.001) following the intervention. In calcified stenoses, the extent of perfusion defects was larger as compared to noncalcified (4.2 +/- 2.5 versus 2.3 +/- 2.0 regions/patient, p < 0.05). CONCLUSION: During rotational atherectomy, myocardial hypoperfusion occurs. The transient nature of this perfusion defect can be demonstrated and quantified by serial 99mTc SPECT. This model may prove useful to assess the effects of pharmacological approaches to reducing myocardial hypoperfusion during coronary rotational atherectomy.
AIM: Of this study was to determine whether success of radioiodine therapy (RIT) in Graves' disease depends on thyroid volume, function, thyroideal receptor antibodies (TRAK), thyreostasis, therapeutic dosage, 131I uptake, or effective half-life. METHOD: 78 patients received an average of 626 +/- 251 MBq of iodine-131 orally for thyroid ablation. 60 were assessed for successful therapy 3 months after RIT. RESULTS: In patients showing hyperthyreosis or a TRAK value > 11 U/l at the beginning of RIT, a significantly lower therapeutic dosage and effective iodine half-life were found than in non-hyperthyreotic patients or patients with TRAK < or = 11 U/l. Patients with a thyroid volume < or = 25 ml showed a significantly lower 131I uptake, but a significantly higher relative uptake (131I uptake/ volume) than patients with a thyroid volume > 25 ml. All failures were treated thyreostatically during RIT and showed a significantly lower therapeutic iodine dosage and relative uptake, as well as a significantly higher thyroid volume than patients with a successful therapy. RIT caused a thyroid volume reduction of 44%, with therapy failures showing a significantly lower volume reduction. Patients who received a therapeutic dosage of < or = 250 Gy showed significantly worse results than did those who had received > 250 Gy. Only one case of therapy failure received a dosage > 250 Gy, while 50% of failures received dosages > 200 Gy but < 250 Gy. Multivariate analyses (MANOVA, factor analyses) showed thyreostasis as the decisive negative factor for a successful course of therapy. CONCLUSIONS: Since most treatment failures occurred in patients under thyreostatic medication we recommend raising the target dosage to 250 Gy for these cases.
AIM: Post injection transmission measurement (PIT) can be performed using rotating 68Ge/68Ga linesources. This study estimates attenuation coefficients, count densities and relative regional uptake values of PIT corrected cardiac PET (E-PIT) compared to routinely pre-injection transmission measurement (RT). METHODS: A thorax-phantom with homogeneously filled myocardium or with simulated defects and six patients with advanced coronary artery disease were studied using ECAT Exact tomograph (Siemens CTI) equipped with three rotating linesources. Transmission was performed twice (PIT, RT), attenuation coefficients and emission data were analysed, the latter without attenuation correction (E-UK), corrected with PIT (E-PIT) and with RT (E-RT) (count density, standard and relative uptake values). RESULTS: Both in phantom and patient studies attenuation coefficients differed significantly between PIT and RT. Comparing E-PIT and E-RT, regional uptake values were different only in phantom simulation with myocardial radioactivity concentrations higher than 10 kBq x ml-1. The image contrast between defects and remaining myocardium in the phantom studies or the standard and relative uptake values in patient studies did not vary significantly. CONCLUSION: Under clinical conditions a post injection transmission measurement does not influence the accuracy of regional myocardial uptake values relevantly.
PURPOSE: The aim of the present study was to determine the acute effects of the "Ecstasy" analogue MDE (3, 4-methylendioxyethamphetamine) on the cerebral glucose metabolism (rMRGlu) of healthy volunteers. METHOD: In a randomised double-blind trial, 16 healthy volunteers without a history of drug abuse were examined with 18-FDG PET 110-120 minutes after oral administration of 2 mg/kg MDE (n = 8) or placebo (n = 8). Beginning two minutes prior to radiotracer injection, a constant cognitive stimulation was maintained for 32 minutes using a word repetition paradigm in order to ensure constant and comparable mental conditions during cerebral 18-FDG uptake. Individual brain anatomy was represented using T1-weighted 3D flash MRI, followed by manual regionalisation into 108 regions-of-interest and PFT/MRI overlay. Absolute quantification of rMRGlu and comparison of glucose metabolism under MDE versus placebo were performed using Mann-Whitney U-test. RESULTS: Absolute global MRGlu was not significantly changed under MDE versus placebo (MDE: 41.8 +/- 11.1 mumol/min/100 g, placebo: 50.1 +/- 18.1 mumol/min/100 g, p = 0.298). The normalised regional metabolic data showed a significantly decreased rMRGlu in the bilateral frontal cortex: left frontal posterior (-7.1%, p < 0.05) and right prefrontal superior (-4.6%, p < 0.05). On the other hand, rMRGlu was significantly increased in the bilateral cerebellum (right: +10.1%, p < 0.05; left +7.6%, p < 0.05) and in the right putamen (+6.2%, p < 0.05). CONCLUSIONS: The present study revealed acute neurometabolic changes under the "Ecstasy" analogon MDE indicating a fronto-striato-cerebellar dysbalance with parallels to other psychotropic substances and various endogenous psychoses respectively.
Photoaffinity labelling of the human poly(ADP-ribose) polymerase (PARP) catalytic domain (40 kDa) with the NAD+ photoaffinity analogue 2-azido-[alpha-32P]NAD+ has been used to identify NAD+-binding residues. In the presence of UV, photo-insertion of the analogue was observed with a stoichiometry of 0.73 mol of 2-azido-[alpha-32P]NAD+ per mol of catalytic domain. Competition experiments indicated that 3-aminobenzamide strongly protected the insertion site. Residues binding the adenine ring of NAD+ were identified by trypsin digestion and boronate affinity chromatography in combination with reverse-phase HPLC. Two major NAD+-binding residues, Trp1014 of peptide Thr1011-Trp1014 and Lys893 of peptide Ile979-Lys893, were identified. The site-directed mutagenesis of these two residues revealed that Lys893, but not Trp1014, is critical for activity. The close positioning of Lys893 near the adenine ring of NAD+ has been confirmed by the recently solved crystallographic structure of the chicken PARP catalytic domain [Ruf, Menissier-de Murcia, de Murcia and Schulz (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 7481-7485].
Peripheral blood stem cells (PBSC) are used for transplantation to reconstitute the hematopoietic system after high-dose chemotherapy. PBSC are harvested from peripheral blood upon successful mobilization by cytokines and/or chemotherapy. Further in vitro manipulation steps like enrichment of CD34+ PBSC or gene transfer can be performed. To ensure the quality and safety of the final cell preparations intended for transplantation, national and international guidelines and regulations have been issued. Herein the implementation of a quality assurance program including the principles of good manufacturing practice (GMP) and a quality control (QC) system is one major concern. GMP regulations apply to all phases of cell collection, processing and storage as well as documentation, training of personnel, and the laboratory facility. QC measures have to be taken to ensure consistent quality and safety with an emphasis on preventing any deficiencies.
To assess dynamic brain function in adults who had stuttered since childhood, regional cerebral blood flow (rCBF) was measured with H2O and PET during a series of speech and language tasks designed to evoke or attenuate stuttering. Speech samples were acquired simultaneously and quantitatively compared with the PET images. Both hierarchical task contrasts and correlational analyses (rCBF versus weighted measures of dysfluency) were performed. rCBF patterns in stuttering subjects differed markedly during the formulation and expression of language, failing to demonstrate left hemispheric lateralization typically observed in controls; instead, regional responses were either absent, bilateral or lateralized to the right hemisphere. Significant differences were detected between groups when all subjects were fluent-during both language formulation and non-linguistic oral motor tasks-demonstrating that cerebral function may be fundamentally different in persons who stutter, even in the absence of stuttering. Comparison of scans acquired during fluency versus dysfluency-evoking tasks suggested that during the production of stuttered speech, anterior forebrain regions-which play an a role in the regulation of motor function-are disproportionately active in stuttering subjects, while post-rolandic regions-which play a role in perception and decoding of sensory information-are relatively silent. Comparison of scans acquired during these conditions in control subjects, which provide information about the sensorimotor or cognitive features of the language tasks themselves, suggest a mechanism by which fluency-evoking maneuvers might differentially affect activity in these anterior and posterior brain regions and may thus facilitate fluent speech production in individuals who stutter. Both correlational and contrast analyses suggest that right and left hemispheres play distinct and opposing roles in the generation of stuttering symptoms: activation of left hemispheric regions appears to be related to the production of stuttered speech, while activation of right hemispheric regions may represent compensatory processes associated with attenuation of stuttering symptoms.
OBJECTIVE: To compare and contrast the patient characteristics of ED patients at low risk for acute cardiac ischemia who were assigned to a chest pain observation service vs those admitted to a monitored inpatient bed for "rule-out acute myocardial infarction" (R/O MI). METHODS: This was a retrospective, cross-sectional comparison of adult patients considered at relatively low risk for cardiac ischemia and who were evaluated in 1 of 2 settings: a short-term observation service and an inpatient monitored bed. All patients had an ED final diagnosis of "chest pain," "R/O MI," or "unstable angina" during the 7-month study period. Demographic features and presenting clinical features were examined as a function of site of patient evaluation. RESULTS: Of 531 study patients, 265 (50%) were assigned to the observation service. Younger age (OR = 1.75, 95% CI 1.26, 2.44, for each decrement of 20 years), the complaint of "chest pain" (OR = 2.35, 95% CI 1.34, 4.12), and the absence of prior known coronary artery disease (OR = 1.64, 95% CI 1.13, 2.38) were the principal independent factors associated with assignment to a chest pain observation service bed. CONCLUSIONS: Patients evaluated in a chest pain observation service appear to have different clinical characteristics than other individuals admitted to a monitored inpatient bed for "R/O MI." Investigators should address differences in clinical characteristics when making outcome comparisons between these 2 patient groups.
UNLABELLED: SPECT/PET studies in schizophrenia revealed inconsistent changes of regional cerebral blood flow (rCBF). Frontal hyperperfusion as well as hypoperfusion are described. This study was undertaken to investigate the relations between rCBF, psychopathology according to PANSS and effects of neuroleptic therapy. METHODS: Twenty-four drug-naive acute patients with a first manifestation of schizophrenia were examined with 99mTc-HMPAO brain SPECT and assessed according to PANSS. Of these, 22 were controlled again after neuroleptic treatment. Following attenuation correction, region-to-cerebellar count ratios were obtained from 98 irregular regions of interest drawn in all slices (6.25 mm). The ratios were compared to 20 control subjects, and changes lying outside of 2 s.d. were considered abnormal. RESULTS: In different drug-naive patients, hyperperfusion as well as hypoperfused patterns were found. In drug-naive patients, the seven subscores of positive symptoms (pos 1-7) in PANSS showed different correlations to rCBF: Formal thought disorders (pos 2) and grandiosity (pos 5) were positively correlated to bifrontal and bitemporal rCBF (r = +0.59 to +0.70). Delusional ideas (pos 1), hallucinatory behavior (pos 3) and suspiciousness (pos 6) demonstrated a negative correlation to bifrontal, cingulate, left temporal and left thalamic rCBF (r = -0.59 to -0.66). Stereotyped ideas (neg 7) as a negative symptom showed a negative correlation to left frontal, left temporal and left parietal rCBF (r = -0.59 to -0.65). No correlations were found between residual positive symptoms and rCBF after neuroleptic treatment and clinical improvement, but all negative symptoms (neg 1-7) had a negative correlation to bifrontal, bitemporal, cingulate, basal ganglia and thalamic rCBF (r = -0.59 to -0.74). CONCLUSION: Our results illustrate that different positive symptoms are accompanied by different rCBF values: some induce hyperperfusion, others hypoperfusion. After therapy (and reduction of positive symptoms), only negative symptoms correlate exclusively to hypoperfusion. This may be the crucial factor in explaining inconsistencies of past results in perfusion pattern in drug-naive schizophrenic patients.
AIM: A new strategy is introduced for a double-head gamma-camera to calculate and to correct for attenuation distribution from count rates of backscattered radiation of 99mTc in the human thorax/myocardium setting. The intention was to gain segmented and calibrated images of distribution employing the structures of lungs and other tissues, so that attenuation may be subsequently corrected by the method of Chang METHODS: Acquisition was done simultaneously in three energy window for 99mTc to receive both emitted and scattered radiation. The acquire projections were used to calculate the attenuation distribution and to execute a scatter correction by the method of Ogawa et. al. By measuring phantoms and evaluating a set of patient studies, quality for both of the approaches towards attenuated distribution and the quality of attenuation correction was tested and an optimal setting for parameters involved was selected. RESULTS: The procedure adequately approaches the correction for both emitted and scattered radiation in SPECT, as experienced in phantom and patient studies. Clinical usefulness was preliminary documented in selected cases. In normal perfusion, distribution was more homogeneous. Perfusion defects appeared more definitive. Problems occurred in high adjacent non-target activity (e.g., in the gallbladder) and with low count rate statistics. CONCLUSION: Myocardial 99mTc MIBI SPECT may be effectively corrected for both attenuation a scatter without an additional transmission device.
UNLABELLED: PET imaging of myocardial perfusion and metabolism identifies regional viability as well as patients at high risk for future cardiac events. This study evaluated a combined "hybrid" imaging approach using 99mTc-sestamibi SPECT and [18F]fluoro-2-deoxy-D-glucose (FDG) PET with regard to reversibility of regional dysfunction and to patient clinical outcome during a 2-yr follow-up. METHODS: In this study, 161 consecutive patients underwent baseline viability imaging. All had regional wall motion (RWM) abnormalities and 88% had a history of previous myocardial infarction. Regions were classified by semiquantitative analysis of sestamibi and FDG uptake as normal, mild match, mismatch or scar. For clinical outcome, patients were divided into three groups: predominantly scar tissue (Group A, n = 90), mild match (Group B, n = 26) and mismatch (Group C, n = 45). Treatment was performed with the knowledge of nuclear results. Cardiac events during follow-up were defined as death, myocardial infarction, unstable angina requiring revascularization, heart transplantation and survived resuscitation. RESULTS: Patients were followed for 29 +/- 6 mo. Revascularization rate was 30% in Group A, 81% in Group B and 80% in Group C, whereas the other patients were treated by medication. Only Group C demonstrated a significant reduction of cardiac events after revascularization, whereas, particularly in Group A, revascularization did not influence the frequency of events. Subjective assessment of angina pectoris and heart failure revealed more patients with improvement after revascularization as compared with conservative treatment. Of the 84 revascularized patients, 61 underwent follow-up angiography at 5 +/- 2 mo with RWM analysis using the centerline method. RWM improved only in mismatch regions from -2.2 +/- 1.0 s.d. to -1.0 +/- 1.4 s.d. (p < 0.01), whereas regions with a mild match or scar did not change. CONCLUSION: Nuclear imaging using 99mTc-sestamibi SPECT and [18F]FDG PET allows diagnosis of viability in regions with reduced perfusion and function with prognostic implications for functional outcome as well as for identification of patients who benefit most from revascularization.
Pheochromocytomas are active, catecholamine-producing tumours derived from chromaffine tissue, causing arterial hypertension. The therapeutical aim is the surgical removal of the tumour. Blockade of the alpha-adrenergic receptors during the preoperative phase is necessary to prevent cardiovascular complications. Phenoxybenzamine is the commonly used alpha-adrenergic blocking agent. Therapy begins with low doses and is increased stepwise up to 250 mg daily. The optimal duration of preoperative therapy is 10 to 14 days. Efficiency of therapy should be judged by reduction of symptoms, a stabilisation of arterial blood pressure, and the presence of light orthostatic hypotension. Swelling of nasal mucosa indicates a successful blockade of the alpha-adrenergic receptors. Persistent tachyarrhythmia is an indication to apply beta-adrenal receptor blockers, but only after alpha-blockade. Reexpansion of blood volume has to be adjusted according to hemodynamic monitoring data. With an adequate preoperative management, mortality of patients with pheochromocytomas has been reduced to less than 1 percent. Postoperatively, most patients become normotensive and other symptoms of excessive catecholamine-production disappear. In cases of long persisting and fixed hypertension paroxysmal crises of high blood pressure can be avoided.