The M-Maze task: An automated method for studying fear memory in rats exposed to protracted aversive conditioning.

BACKGROUND: Fear conditioning (FC) in rodents is the most used animal model to investigate the neurobiology of posttraumatic stress disorder (PTSD). Although research using FC has generated a better understanding of fear memories, studies often rely on mild or moderate FC training and behavioral analysis generally focuses on measuring freezing responses within few test sessions. NEW METHOD: We introduce the M-Maze task, a system that measures extinction of conditioned fear using suppression of operant behavior. The apparatus consists of an M-shaped maze where rats are trained to alternate nose poking at two pellet dispensers. Proximity sensors measure the animal's locomotion, as well as the latencies and number of operant behaviors. Here we also describe the protracted aversive conditioning (PAC), a rat model of severe fear that induces resistant extinction following a 4-day conditioning protocol that combines delay, unpredictable, and short- and long-trace conditioning. RESULTS: An intense one-day auditory FC protocol induced a sharp elevation in transit time and suppression of nose pokes by conditioned cues, but in contrast to what is found in PTSD patients, fear extinction was rapidly observed. On the other hand, PAC alone or in combination with exposure to single prolonged stress induced persistent extinction impairments in M-Maze tests, as well as enhanced anxiety, and social withdrawal. COMPARISON WITH OTHER EXISTING METHODS: The M-Maze task is fully automated and allows multiple animals to be tested simultaneously in long-term experiments. Moreover, PAC training can be an alternative approach to study extinction-resistant fear. CONCLUSIONS: The M-Maze task allows rapid and unbiased measurements of fear-induced suppression. We suggest that long-term assessment of extinction impairments would lead to a better understanding of the neurobiology of persistent fear and the screening for new therapies.

Factor XIII cotreatment with hemostatic agents in hemophilia A increases fibrin α-chain crosslinking.

Essentials Factor XIII (FXIII)-mediated fibrin crosslinking is delayed in hemophilia. We determined effects of FXIII cotreatment with hemostatic agents on clot parameters. FXIII cotreatment accelerated FXIII activation and crosslinking of fibrin and α2 -antiplasmin. These data provide biochemical rationale for FXIII cotreatment in hemophilia. SUMMARY: Background Hemophilia A results from the absence, deficiency or inhibition of factor VIII. Bleeding is treated with hemostatic agents (FVIII, recombinant activated FVII [rFVIIa], anti-inhibitor coagulation complex [FEIBA], or recombinant porcine FVIII [rpFVIII]). Despite treatment, some patients have prolonged bleeding. FXIII-A2 B2 (FXIII) is a protransglutaminase. During clot contraction, thrombin-activated FXIII (FXIIIa) crosslinks fibrin and α2 -antiplasmin, which promotes red blood cell retention and increases clot stability and weight. We hypothesized that FXIII cotreatment in hemophilia would accelerate FXIII activation, leading to increased fibrin crosslinking. Methods FVIII-deficient plasma and whole blood were clotted with or without hemostatic agents (FVIII, rFVIIa, FEIBA, or recombinant B-domain-deleted porcine FVIII [rpFVIII]) and/or FXIII. The effects on FXIII activation, thrombin generation, fibrin and α2 -antiplasmin crosslinking, clot formation and clot weight were measured by western blotting, calibrated automated thrombography, thromboelastography, and clot contraction assays. Results As compared with FVIII-treated hemophilic plasma, FVIII + FXIII cotreatment accelerated FXIIIa formation without increasing thrombin generation. As compared with buffer-treated or FXIII-treated hemophilic plasma, FVIII treatment and FVIII + FXIII cotreatment increased the generation and amount of crosslinked fibrin, including α-chain-rich high molecular weight species and crosslinked α2 -antiplasmin. In the presence of FVIII inhibitors, as compared with hemostatic treatments (rFVIIa, FEIBA, or rpFVIII) alone, FXIII cotreatment increased whole blood clot weight. Conclusion In hemophilia A plasma and whole blood, FXIII cotreatment with hemostatic agents accelerated FXIIIa formation, increased the generation and amount of fibrin α-chain crosslinked species, accelerated α2 -antiplasmin crosslinking, and increased clot weight. FXIII cotreatment with hemostatic therapy may augment hemostasis through increased crosslinking of fibrin and α2 -antiplasmin.

Virtual reality hardware and graphic display options for brain-machine interfaces.

Virtual reality hardware and graphic displays are reviewed here as a development environment for brain-machine interfaces (BMIs). Two desktop stereoscopic monitors and one 2D monitor were compared in a visual depth discrimination task and in a 3D target-matching task where able-bodied individuals used actual hand movements to match a virtual hand to different target hands. Three graphic representations of the hand were compared: a plain sphere, a sphere attached to the fingertip of a realistic hand and arm, and a stylized pacman-like hand. Several subjects had great difficulty using either stereo monitor for depth perception when perspective size cues were removed. A mismatch in stereo and size cues generated inappropriate depth illusions. This phenomenon has implications for choosing target and virtual hand sizes in BMI experiments. Target-matching accuracy was about as good with the 2D monitor as with either 3D monitor. However, users achieved this accuracy by exploring the boundaries of the hand in the target with carefully controlled movements. This method of determining relative depth may not be possible in BMI experiments if movement control is more limited. Intuitive depth cues, such as including a virtual arm, can significantly improve depth perception accuracy with or without stereo viewing.

Granisetron vs ondansetron for prevention of nausea and vomiting in hematopoietic stem cell transplant patients: results of a prospective, double-blind, randomized trial.

The serotonin type-3 (5-HT3) antagonists represent a significant advance in the prevention of acute nausea and vomiting (N/V) from highly emetogenic chemotherapy. We sought to determine if any differences in efficacy or adverse effects exist between two such agents, ondansetron and granisetron, during conditioning therapy for hematopoietic stem cell transplantation (HSCT). Patients were randomized to receive either ondansetron 0.15 mg/kg intravenously every 8 h or granisetron 10 microg/kg intravenously daily. Additionally, all patients received scheduled dexamethasone and lorazepam. Prophylaxis was continued until 24 h after completion of chemotherapy. Nausea and distress were measured subjectively with visual analog scales and emetic episodes were quantified. Of the 110 randomized patients, 96 were evaluable for efficacy and safety. No significant differences in efficacy were observed between the ondansetron- and granisetron-treated patients, evaluated by comparing the degree of nausea and distress, number of emetic episodes and overall control of emesis. The adverse effects were also comparable and no patients were removed from study because of severe toxicities. This trial demonstrates that ondansetron and granisetron are equally effective at preventing acute N/V associated with conditioning therapy frequently used for HSCT. The agent of choice should be based on drug acquisition cost or preference.

In vivo studies of the anti-tumor effects of a human prolactin antagonist, hPRL-G129R.

Previously we demonstrated that a mutated human prolactin (hPRL) with a single amino acid substitution at position 129 (hPRL-G129R) was able to inhibit human breast cancer cell proliferation via the induction of apoptosis. In this study, we report the in vivo anti-tumor effects of hPRL-G129R in nude mice bearing human breast cancer xenografts (T-47D and MCF-7). In an effort to prolong the half-life of the proteins, hPRL or hPRL-G129R were formulated with either growth factor reduced Matrigel or into slow-releasing pellets (custom made 5 mg/5 day release). Initially, nude mice inoculated (s.c.) with T-47D human breast cancer cells were treated with either hPRL or hPRL-G129R formulated with Matrigel. At the end of the 7-week study, it was found that hPRL significantly stimulated the in vivo growth of T-47D xenografts (mean tumor volume, 202 +/- 62 mm(3) as compared to 124 +/- 31 mm(3) in control mice), whereas hPRL-G129R inhibited the tumor growth (mean tumor volume, 79+/-32 mm3). The inhibitory effects of hPRL-G129R were further confirmed in a second experiment using nude mice bearing MCF-7 human breast cancer xenografts and treated with slow-releasing pellets containing hPRL-G129R. Based on these results, we believe that hPRL-G129R can be used to improve the outcome of human breast cancer treatment in the near future.

Combination of PCR subtraction and cDNA microarray for differential gene expression profiling.

PCR subtraction hybridization has been used effectively to enrich and single out differentially expressed genes. However identification of these genes by means of cloning and sequencing individual cDNAs is a tedious and lengthy process. In this report, an attempt has been made to combine the use of PCR select cDNA subtraction hybridization and cDNA microarrays to identify differentially expressed genes using a nonradioactive chemiluminescent detection method. mRNA from human prolactin (hPRL) or human prolactin antagonist (hPRL-G129R) treated and non-treated breast cancer cells was isolated, and cDNAs were synthesized and used for the PCR subtraction to enrich the differentially expressed genes in the treated cells. The PCR-amplified and subtracted cDNA pools were purified and labeled using the digoxigenin method. Labeled cDNAs were hybridized to a human apoptosis cDNA microarray membrane and identified by chemiluminescence. The results suggest that the strategy of combining all three methods will allow for a more efficient, nonradioactive way of identifying differentially expressed genes in target cells.

[Effect of iodine and thyroid hormones in the induction and therapy of Hashimoto's thyroiditis]. / Effekte von Iodid und Schilddrüsenhormonen bei der Induktion und Therapie einer Thyreoiditis Hashimoto.

AIM: The effect of an iodine prophylaxis on the induction of Hashimoto's thyroiditis as well as the influence of various therapeutic approaches on the course of antithyroglobulin (TgAb) and antiperoxidase (TPOAb) antibodies in manifest diseases are evaluated. METHOD: A collective of 375 euthyroid subjects without relevant goiter received daily doses of 200 micrograms iodide, weekly doses of 1.53 milligrams iodide, or no medication. A second group of 377 patients suffering from Hashimoto's thyroiditis was treated with a non-suppressive hormone medication, a suppressive hormone administration, a combination of a non-suppressive hormone therapy with low dose iodide (50-150 micrograms/day), mere iodide in doses of 200 micrograms/day, or received no therapy. The mean observation period in these two groups was 860 and 848 days, respectively. RESULTS: There was no significant increase of the antibody levels in the subgroup with 200 micrograms iodide/day and in the non-treated subjects of the first collective. However, the group that received 1.53 milligrams iodide/week presented a distinct increase of the TgAb as well as the TPOAb, and the incidence of Hashimoto's thyroiditis was 4-fold higher than in the two other subgroups. The patients of the second collective revealed a significant decrease of the TgAb in the subgroups treated with up to 200 micrograms iodide/day, while the reduction of the TPOAb depended on the thyrotropin level and was most significant in the suppressed group (p < 0.0001). CONCLUSION: To lower the incidence of autoimmune thyroid diseases in predisposed subjects, a daily iodine supplementation seems to be superior to high-dose weekly administrations. A hormone therapy combined with a daily, low-dose iodine medication is able to reduce the TgAb and the TPOAb levels even in patients with Hashimoto's thyroiditis.

Successful PBSC mobilization with high-dose G-CSF for patients failing a first round of mobilization.

PBSC are the preferred source of stem cells for autologous transplantation. However, regardless of the mobilization procedure used, 10%-20% of patients fail to collect an adequate number to ensure prompt engraftment. There is as yet no standard mobilization procedure for patients who fail a first mobilization attempt. Here, we describe a highly efficient strategy to obtain an adequate number of stem cells for patients who failed a first mobilization attempt. Seventy-four patients with various hematologic malignancies underwent initial mobilization with various regimens including hematopoietic growth factors with or without chemotherapy. In 72% of patients, > or =2 x 10(6) CD34+ stem cells/kg were collected in the initial mobilization attempt, and patients engrafted in a median of 10 days for neutrophils and 12 days for platelets. Eighteen patients failed to mobilize adequate numbers of stem cells, defined as the inability to collect 0.2 x 10(6) CD34+ stem cells/kg/day in the first 2-3 days. These patients had their apheresis halted. Patients were immediately given G-CSF (32 microg/kg/day) for 4 days as a second attempt at mobilization. Eighty-eight percent of these patients achieved the target of > or =2 x 10(6) CD34+ cells/kg, with a median duration of apheresis of 5 days (including the first and second mobilizations). The mean CD34+ cells/kg/day increased after administration of high-dose G-CSF from 0.16 after the first mobilization attempt to 0.61 (p = 0.0002) after the second mobilization. All patients engrafted in a median of 11 and 13 days for neutrophils and platelets, respectively. We conclude that patients whose apheresis yield is <0.4 x 10(6) CD34+ cells/kg after the first two apheresis collections can be successfully mobilized if high-dose G-CSF is administered immediately and continued until achieving > or =2 x 10(6) CD34+ stem cells/kg.

[Individual calculation of the minimal effective levothyroxine dose in prolonged suppression tests]. / Individuelle Bestimmung der minimal erforderlichen Levothyroxindosis bei Langzeit-Suppressionstests.

AIM: A formula for calculating the minimum suppressive levothyroxine dose in prolonged suppression tests using body weight and TSH level is derived from a large number of cases. METHOD: In 1239 euthyroid patients (TSH > 0.30 mU/l) a suppression test with levothyroxine substitution for at least 6 weeks was performed. The hormone dose was primarily based on the initial TSH level using the empirical value TSH x 100, with a range between 50 and 150 micrograms per day. RESULTS: In 900 patients (73%), the TSH level was suppressed during the medication (TSH < 0.30 mU/l). Among them, we found 211 cases with an intermediate TSH suppression (TSH between 0.10 and 0.20 mU/l). Following the formula f = levothyroxine dose [microgram]/(body weight [kg] x initial TSH level [mU/l]) an average factor of 1.57 was calculated to obtain the suppressive dose in this group. As usually a complete TSH suppression (< 0.10 mU/l) is required, we recommend factor 2 for practical use. A consideration of the body weight revealed that 313 patients actually received a hormone dose equivalent to 2 x weight x TSH. In this group, a TSH suppression was found in 262 patients (84%), a factitial hyperthyreosis was not observed. When contemplating the three subgroups of this collective, who received daily doses of 50, 75 and 100 micrograms levothyroxine, respectively, we neither found a significant difference in the average body weight, nor in the value of the suppressed TSH. However, the correlation between the average initial TSH levels in these groups and the respective suppressive hormone doses clearly demonstrates their dependence on the thyroid regulation. CONCLUSION: The minimum suppressive levothyroxine dose does not only depend on the body weight, but also on the initial TSH level. It can be estimated using the formula 2 x body weight x initial TSH (range between 50 and 150 micrograms per day).

[Individual outcome monitoring of TSH receptor antibody level for predicting the prognosis of Graves' disease]. / Individuelle Verlaufskontrolle der TSH-Rezeptor-Antikörper-Titer zur Einschätzung der Prognose eines M. Basedow.

AIM: This retrospective study is intended to provide further information on the controversially discussed ability of a Thyrotropin Receptor Antibody (TRAb) monitoring for predicting the outcome of Graves' disease. METHOD: The study is based on 1480 blood samples of 346 patients (292 female, 54 male; age among 9 and 91 years) suffering from Graves' disease. A comparison between the TRAb levels and the serum free thyroid hormones as well as the basal thyrotropin was performed in the entire collective, in the group of non-treated patients and in 182 individual courses. RESULTS: Even in comparable states of function, the individual TRAb levels were quite divergent. In the group with persistent dysfunction, they initially varied between 6 and 482 U/l (normal range < 15 U/l). A global evaluation of the particular groups hence did not show any correlation between the functional disorder and the respective TRAb levels. However, when considering the individual courses, a distinct dependence upon the development of the TRAb could be observed. As the limit for separating the group suffering from persistent malfunction or recurrence and the collective in remission, a decline of the individual TRAb level below 50% of the initial value has proven to be most suitable. This was observed in 94% of all remissions. In 90% of all relapsed patients, we either found persistent high TRAb levels (i.e. a maximum decrease down to 50% of the initial value) or another increase of the TRAb levels. The positive and negative predictive values of the method are 97% and 58%, respectively. CONCLUSION: The outcome of Graves' disease depends on the individual changes of the TRAb levels rather than on their absolute values. In case of persistent TRAb, you have to expect a continuing dysfunction or a recurrence.

Pegaspargase: an alternative?

OBJECTIVE: To review the chemistry, pharmacology, pharmacokinetics, clinical activity, adverse effects, dosage, and administration guidelines for pegaspargase. DATA SOURCES: A MEDLINE search (1980-1996), a CANCERLIT search (1983-1996), and a CURRENT CONTENTS search (1980-1996) using the terms pegaspargase, PEG-asparaginase, PEG-L-asparaginase, polyethylene glycol L-asparaginase, polyethylene glycol conjugated L-asparaginase, and Oncaspar were conducted. STUDY SELECTION AND DATA EXTRACTION: All articles were considered for possible inclusion in this review. Abstracts were included only when they were judged to add critical information not otherwise available in the medical literature. DATA SYNTHESIS: L-Asparaginase has been a main component of treatment regimens for acute lymphocytic leukemia. A key limiting factor of L-asparaginase use has been the development of hypersensitivity to the drug. Recently, a polyethylene glycol (PEG) conjugated form of L-asparaginase, pegaspargase, has been made available. PEG modification of L-asparaginase has been shown to alter the tendency of the enzyme to induce an immune response and to extend the half-life of the drug. The majority of patients with hypersensitivity to the native enzyme preparations tolerate pegaspargase without further clinical hypersensitivity. The adverse effect profile of pegaspargase is similar to that of the native forms of L-asparaginase. The recommended dosage of pegaspargase is 2500 IU/m2 administered by intramuscular or intravenous injection every 2 weeks in combination with other chemotherapeutic agents. CONCLUSIONS: Pegaspargase is a safe, effective alternative to L-asparaginase in patients who have had clinical hypersensitivity reactions to both Escherichia coli- and Erwinia carotovora-derived L-asparaginase. However, pegaspargase should not be routinely substituted for L-asparaginase.

[Palliative treatment for pain in osseous metastasized prostatic carcinoma with osteotropic rhenium-186 hydroxyethylidene diphosphonate (HEDP)]. / Palliative Schmerztherapie beim ossär metastasierten Prostatakarzinom mit dem osteotropen beta-Strahler 186Rhenium-Hydroxyethylidendiphosphonat (HEDP).

Systemic administration of strontium-89 is an important option for pain relief in advanced prostate carcinoma with multiple osseous metastases. Recently, rhenium-186-HEDP was introduced as a new substance which has important advantages (shorter physical half-life, scintigraphic imaging, dose distribution). The myelosuppressive effect can be estimated more accurately in advance, so that adverse effects can be reduced and the treatment can be repeated after a shorter period of time and more often. Our study comprises 15 treatments with rhenium-186-HEDP in advanced prostate cancer patients using the 1.4- to 2-fold standard dose. The response rate, estimated as reduction in pain and increase in patient mobility, was 87% with no major myelosuppressive effects. The mean duration of pain relief was 4-6 weeks. All four patients with repeated therapy were also responding to the second treatment. Radionuclide therapy for painful osseous metastases with rhenium-186-HEDP appears to be an effective and, even at higher doses, safe procedure.

Investigations of breast tumors with fluorine-18-fluorodeoxyglucose and SPECT.

METHODS: We designed a prospective study to investigate the feasibility of combined FDG-SPECT and whole-body acquisition in the diagnostic work-up of breast tumors applying visual analysis. We studied 50 patients with breast tumors of unknown histology. RESULTS: All malignant diseases were accurately detected in tumors > 2.3 cm, while the smallest FDG-positive lesion was 1.4 cm. In a subgroup of these patients, quantitative evaluation (tumor-to-back-group ratios) was added, which improved the sensitivity. Lymph node metastases were accurately indicated in 9 of 13 patients, while the detection of distant metastases depended on the location and size. False-positive FDG scans were observed in inflamed tissue, in a rapidly growing phylloides tumor and in supposedly healthy breasts. CONCLUSION: These results are comparable with prior investigations of other groups using PET. Therefore, FDG-SPECT and whole-body acquisition may be an adequate and less expensive technique to meet the increasing demand of FDG examinations.

Valproate in status epilepticus.

In patients with intractable status epilepticus who have not responded to therapy with benzodiazepines, phenytoin, and barbiturates, valproate may be a reasonable option. Extemporaneously prepared valproate rectal suppositories or retention enemas have been given in dosages of 200-1200 mg q6h in addition to phenytoin, phenobarbital, or both in adults. The pediatric dose used was 15-20 mg/kg, in addition to phenytoin and/or phenobarbital.

[18FDG in the primary staging of lung tumors. Results with a gamma camera and a 511 keV collimator]. / 18FDG beim primären Staging von Lungentumoren. Ergebnisse an einer Gammakamera mit einem 511 keV Kollimator.

The diagnosis of primary lung tumors requires a precise staging according to the TNM classification. In contrast to established imaging methods 18FDG describes the functional metabolic processes in the tumor tissue due to increased glycolysis. This paper describes the use of 18FDG in the primary staging of lung tumors and metastases. 44 patients were studied with a gamma camera and a 511 keV collimator. In comparison to pulmonary tumors and metastases detected by other imaging methods (107) the accumulation of 18FDG has a sensitivity of 85%, in lesions verified by histology (50) of 89%, in primary tumors (35) of 100% and in metastases (63) of 76%. As an alternative to FDG PET studies, primary staging of lung tumors is possible with a gamma camera, suitable for ECT and fitted with a 511 keV collimator.

Chronopharmacokinetics of imipramine and desipramine in rat forebrain and plasma after single and chronic treatment with imipramine.

Daily variations in the pharmacokinetics of imipramine (IMI) could contribute to circadian phase-dependent effects of the drug. Therefore, the chronopharmacokinetics of IMI and its metabolite, desipramine (DMI), were studied after single and chronic application. Male rats were synchronized to a 12:12 hour light:dark (L:D) regimen with lights on from 07:00 to 19:00 (dark, 19:00-07:00). In single-dose experiments rats were injected with IMI (10 mg/kg) i.p. or i.v. at 07:30 or 19:30 and groups of rats were killed 0-22 hours thereafter. After chronic application of IMI in drinking water (approximately 15 mg/kg/d) groups of rats were killed during the 14th day of treatment at 02:00, 08:00, 14:00, and 20:00, respectively. Brain and plasma concentrations of IMI and DMI were determined by reversed-phase high-performance liquid chromatography with ultraviolet detection. After single i.p. application of IMI, maximal brain concentrations (Cmax) of IMI and DMI were nearly twofold higher in darkness (IMI, 4.8 micrograms/g; DMI, 1.8 micrograms/g) than in light (IMI, 2.85 micrograms/g; DMI, 0.85 microgram/g). Also, the area under the curve (AUC) (0-22 hours) was about 1.6-fold greater in darkness than in light for IMI and DMI; half-lives were not circadian phase dependent. After i.v. injection of IMI, the AUC in brain was also about 30% greater in darkness than in light. After chronic application of IMI in drinking water, brain concentrations of IMI and DMI varied more than threefold within 24 hours. The data demonstrate that the pharmacokinetics of IMI and DMI are circadian phase dependent. It is assumed that circadian variations in drug distribution are more likely to contribute to the drug's chronopharmacokinetics than variations in the drug's metabolism. The 24-hour variations in the drug's concentrations after chronic IMI application in drinking water can be explained by the drinking behavior of the rats, which by itself is altered by IMI.