Evaluation of 177Lu[Lu]-CHX-A″-DTPA-6A10 Fab as a radioimmunotherapy agent targeting carbonic anhydrase XII.

INTRODUCTION: Due to their infiltrative growth behavior, gliomas have, even after surgical resection, a high recurrence tendency. The approach of intracavitary radioimmunotherapy (RIT) is aimed at inhibiting tumor re-growth by directly administering drugs into the resection cavity (RC). Direct application of the radioconjugate into the RC has the advantage of bypassing the blood-brain barrier, which allows the administration of higher radiation doses than systemic application. Carbonic anhydrase XII (CA XII) is highly expressed on glioma cells while being absent from normal brain and thus an attractive target molecule for RIT. We evaluated a CA XII-specific 6A10 Fab (fragment antigen binding) labelled with 177Lu as an agent for RIT. METHODS: 6A10 Fab fragment was modified and radiolabelled with 177Lu and characterized by MALDI-TOF, flow cytometry and radio-TLC. In vitro stability was determined under physiological conditions. Biodistribution studies, autoradiography tumor examinations and planar scintigraphy imaging were performed on SCID-mice bearing human glioma xenografts. RESULTS: The in vitro CA XII binding capacity of the modified Fab was confirmed. Radiochemical purity was determined to be >90% after 72 h of incubation under physiological conditions. Autoradiography experiments proved the specific binding of the Fab to CA XII on tumor cells. Biodistribution studies revealed a tumor uptake of 3.0%ID/g after 6 h and no detectable brain uptake. The tumor-to-contralateral ratio of 10/1 was confirmed by quantitative planar scintigraphy. CONCLUSION: The radiochemical stability in combination with a successful in vivo tumor uptake shows the potential suitability for future RIT applications with the 6A10 Fab.

Developmental exposure to the SSRI citalopram causes long-lasting behavioural effects in the three-spined stickleback (Gasterosteus aculeatus).

Selective Serotonin re-uptake inhibitors (SSRIs) are a class of psychotropic drugs used to treat depression in both adolescents and pregnant or breast-feeding mothers as well as in the general population. Recent research on rodents points to long-lasting behavioural effects of pre- and perinatal exposure to SSRIs which last into adulthood. In fish however, studies on effects of developmental exposure to SSRIs appears to be non-existent. In order to study effects of developmental SSRI exposure in fish, three-spine sticklebacks were exposed to 1.5 µg/l of the SSRI citalopram in the ambient water for 30 days, starting two days post-fertilisation. After approximately 100 days of remediation in clean water the fish were put through an extensive battery of behavioural tests. Feeding behaviour was tested as the number of bites against a piece of food and found to be increased in the exposed fish. Aggression levels were measured as the number of bites against a mirror image during 10 min and was also found to be significantly increased in the exposed fish. Novel tank behaviour and locomotor activity was tested in an aquarium that had a horizontal line drawn half-way between the bottom and the surface. Neither the latency to the first transition to the upper half, nor the number of transitions or the total time spent in the upper half was affected by treatment. Locomotor activity was significantly reduced in the exposed fish. The light/dark preference was tested in an aquarium where the bottom and walls were black on one side and white on the other. The number of transitions to the white side was significantly reduced in the exposed fish but there was no effect on the latency to the first transition or the total time spent in the white half. The results in the current study indicate that developmental SSRI exposure causes long-lasting behavioural effects in fish and contribute to the existing knowledge about SSRIs as environmental pollutants.

Waterborne citalopram has anxiolytic effects and increases locomotor activity in the three-spine stickleback (Gasterosteus aculeatus).

Citalopram is an antidepressant drug, which acts by inhibiting the re-uptake of serotonin from the synaptic cleft into the pre-synaptic nerve ending. It is one of the most common drugs used in treatment of depression, it is highly lipophilic and frequently found in sewage treatment plant effluents and surface waters around the world. Citalopram and other selective serotonin re-uptake inhibitors have, at concentrations that occur in nature, been shown to have behavioural as well as physiological effects on fish and other animals. This study is the result of several different experiments, intended to analyse different aspects of behavioural effects of chronic citalopram exposure in fish. Our model species the three-spine stickleback is common in the entire northern hemisphere and is considered to be a good environmental sentinel species. Female three-spine sticklebacks were exposed to 0, 1.5 and 15µg/l nominal concentrations of citalopram for 21 days and subjected to the novel tank (NT) diving test. In the NT test, the fish exposed to 1.5µg/l, but not the 15µg/l fish made a significantly higher number of transitions to the upper half and stayed there for significantly longer time than the fish exposed to 0µg/l. The 15µg/l group, however, displayed a significantly lower number of freeze bouts and a shorter total freezing time. The test for locomotor activity included in the NT test showed that fish treated with 1.5 and 15µg/l displayed a significantly higher swimming activity than control fish both 5-7 and 15-17min after the start of the experiment. In the next experiment we compared fish exposed to 1.5µg/l and 0.15µg/l to pure water controls with regard to shoaling intensity and found no effect of treatment. In the final experiment the propensity of fish treated with 1.5µg/l to approach an unknown object and aggressive behaviour was investigated using the Novel Object test and a mirror test, respectively. The exposed fish ventured close to the unknown object significantly more often and stayed there for significantly longer time than unexposed fish. The aggression test yielded no statistically significant effects. It is concluded that citalopram changes the behaviour of the three-spine stickleback in a way that is likely to have ecological consequences and that it must not be considered an environmentally safe pharmaceutical.

Does Pregnenolone Enhance Exposure Therapy in Obsessive-Compulsive Disorder? - A Pilot, Interim Report of a Randomized, Placebo-Controlled, Double-Blind Study.

Exposure therapy is an effective cognitive-behavioral treatment for patients with obsessive-compulsive disorder (OCD). However, a further amelioration of symptoms by additional drugs that enhance extinction learning is desirable. An interesting candidate is pregnenolone, which positively modulates NMDA and GABAA receptors in preclinical studies and influences amygdala and prefrontal activity in humans. We present pilot data showing high acceptance and good tolerability of pregnenolone given 2 h before exposure sessions in OCD patients. As per our interim analyses, exposure treatment resulted in significantly improved main outcome parameters, but no effects of pregnenolone vs. placebo pretreatment were detectable thus far.

High Estradiol Levels During Postmenopause - Pitfalls in Laboratory Analysis.

A 54-year-old woman was admitted with a result of high serum estradiol levels (> 4300 pg/ml) and typical postmenopausal symptoms. She had a history of an adnexectomy (normal histopathology) due to the elevated estradiol levels. After surgery, estradiol levels were as high as before. Analyzing the anti-mullerian hormone (AMH), inhibin B, DHEA-S and estrone, typical postmenopausal levels were found. Serum estradiol levels were controlled several times with rabbit-derived polyclonal as well as monoclonal antibodies to optimize the selectivity of the test system. Secondary, a radioimmunoassay was performed to exclude interferences of the detection system where lower, but still elevated estradiol levels (186 pg/ml) were found. Hypothesizing that our patient underwent a cross reaction with irregular antibodies, a control was done using sheep-derived antibodies, which proved a postmenopausal hormone level (estradiol level < 5 pg/ml). This result was confirmed using a fluorescence enzyme immunoassay (FEIA) revealing high levels of irregular antibodies (> 200 mg/l; reference < 30 mg/l). This case depicts the pitfalls of estradiol measurement detecting false elevated estradiol levels in a postmenopausal woman.

Environmentally relevant concentrations of citalopram partially inhibit feeding in the three-spine stickleback (Gasterosteus aculeatus).

Selective Serotonin Re-uptake Inhibitors (SSRI) are mood-altering, psychotropic drugs commonly used in the treatment of depression and other psychological illnesses. Many of them are poorly degraded in sewage treatment plants and enter the environment unaltered. In laboratory studies, they have been demonstrated to affect a wide range of behaviours in aquatic organisms. In this study we investigated the effect of a three-week exposure to 0.15 and 1.5 µg/l of the SSRI citalopram dissolved in the ambient water on the feeding behaviour in three-spine stickleback (Gasterosteus aculeatus). Feeding, measured as the number of attacks performed on a piece of frozen bloodworms during a 10-min period, was reduced by 30-40% in fish exposed to both 0.15 and 1.5 µg/l citalopram. The effects of the environmentally relevant concentration 0.15 µg/l on feeding, an important fitness characteristic, suggests that the ecological significance of environmental SSRI exposure may be pronounced.

[Severe degloving injury to both feet in a child]. / Schwere Degloving-Verletzung der Füße bei einem Kind.

Although rarely described, it is generally accepted that degloving injuries can successfully be treated by defatting the degloved skin and replacing it with a full-thickness graft. There have been few reports on the outcome of this reconstructive procedure in children. In this report, we describe the use of the VAC system to treat a case of degloving injury to both feet of a 4-year-old boy who was pulled into a luggage belt while standing on it.

[Peripartum hysterectomy--an audit in Slovakia in 2007]. / Peripartálne hysterektómie--audit na Slovensku v roku 2007.

OBJECTIVE: An audit was performed to assess the number, indications and complications of peripartum hysterectomy at the departments of obstetrics/gynecology in Slovakia in 2007. DESIGN: Observational descriptive study. SETTING: Department of Obstetrics and Gynecology, Faculty Hospital Nitra and Constantine the Philosopher University Nitra. METHOD: An official questionnaire of Slovak Society of Obstetrics and Gynecology was sent to all 63 departments of obstetrics/gynecology to find the number of peripartum hysterectomy performed in the year 2007. Differences between intrapartum and postpartum cases were compared. RESULTS: 44 from 63 institutions responded to the survey (response rate 69.8%). There were 38,485 deliveries and 24 cases of peripartum hysterectomies. The incidence of peripartum hysterectomy was 0.62/1000 deliveries, 1 case occurred in 1604 deliveries. 16 (66.7%) patients had a total abdominal hysterectomy with the remaining 8 (33.3%) having a sub-total hysterectomy. All operations were emergent. 18 procedures were performed during delivery and 6 in the postpartum period. Hypogastric artery ligation before hysterectomy were performed on 2 patients in the postpartum group. 20 of 24 (83.3%) patients delivered by cesarean section, three (12.5%) by spontaneous vaginal delivery and one (4.2%) with vaccumextraction. The indications for emergency peripartum hysterectomy were: placenta praevia 6 cases (25%), placental abruption with disseminated intravascular coagulation 6 (25%), placenta accreta 3 (12.5%), uterine atony 3 (12.5%), uterine rupture 3 (12.5%) and retroperitoneal haematoma 3 (12.5%). The youngest patient was 15 year-old, the oldest one was 39. After hysterectomy 10 (41.7%) women were admitted to the intensive care unit. There was no maternal mortality, but five newborns died due to perinatal asphyxia. There were more blood transfusions in the group of postpartum hysterectomies in comparison with intrapartum cases (4.0 +/- 1.3 transfusion units vs 9.1 +/- 3.5, p < 0.05), as well as the longer hospital stay (10.3 +/- s4.2 days vs. 19.1 +/- 5.3, p < 0.05). CONCLUSIONS: Peripartal hysterectomy is a dramatic but a life saving procedure. It is usually associated with significant maternal and fetal morbidity and mortality. Every obstetric service should have access to a surgical team capable of performing emergency peripartal hysterectomy.

[Vacuum therapy--basics, indication, contraindication and cost listing]. / Vakuumversiegelung--Grundlagen, Indikationen, Kontraindikationen und Kostenbilanz.

Vacuum therapy is an efficient alternative to conservative wound care and creates new possibilities for the treatment of a variety of chronic and acute wounds. Already more than two thousand years ago vacuum treatment was applied in traditional Chinese medicine and also today vacuum therapy is used more and more frequently in daily clinical practice. The positive effects of the therapy are based on the stimulation of blood flow, formation of granulation tissue, angiogenesis, as well as cell proliferation, accelerated secondary wound closure and the removal of bacteria from the wound. As can be demonstrated based on a clinical picture of the open abdomen, vacuum therapy based on KCI's (Kinetic Concepts Inc) VA.C. system (Vacuum Assisted Closure) provides a possibility for not only accelerating the healing process, but also significantly lowering mortality. Since vacuum therapy in the form of the V.A.C. system represents a relatively expensive intervention, a comparison with conventional healing methods is demonstrated. Clinical studies with high evidence level show that both economic and clinical advantages of the vacuum therapy speak for its application. Savings with respect to the faster healing rates, the reduced dressing changes, as well as hospital stay and the care requirements, and the reduction of subsequent illnesses, compensate for the higher initial costs. Although many reports exist on treatment successes with the application of V.A.C. therapy, there are several cases for which vacuum therapy is contraindicated. In order to make sure that V.A.C. therapy is effective, a range of precautions and contraindications are described. The European Wound management Association (EWMA) just wrote a position paper on V.A.C. therapy. This is available at www.ewma.org.

[The use of recombinant factor VIIa on the therapy of massive bleeding after cesarean section]. / Pouzitie rekombinantného faktora VIIa na liecbu masívneho krvácania po cisárskom reze.

OBJECTIVE: To present a case report of severe post-partum bleeding after caesarean section and successful treatment using recombinant factor VIIa. SETTING: Obstetrics and Gynecology Department, Faculty Hospital, Nitra, Slovak Republic. CASE REPORT: A 29-year old secundigravida presented with major post-partum bleeding after caesarean section due to uterine atony. The patient developed hemorrhagic shock, associated with disseminated intravascular coagulation (DIC). Treatment with uterotonic drugs, prostaglandins and hysterectomy failed to control diffuse bleeding. Recombinant factor VIIa (90 microg/kg intravenous injection) was given as a final attempt to control the bleeding. The response to treatment was rapid, with control of the bleeding and resolution of the coagulopathy. CONCLUSION: This case suggests a potential role of recombination factor VIIa in the treatment of severe post-partum bleeding associated with DIC.

Efficient and fast targeted production of murine models based on ENU mutagenesis.

Mice with targeted genetic alterations are the most effective tools for deciphering organismal gene function. We generated an ENU-based parallel C3HeB/FeJ sperm and DNA archive characterized by a high probability to identify allelic variants of target genes as well as high efficiencies in allele retrieval and model revitalization. Our archive size of over 17,000 samples contains approximately 340,000 independent alleles (20 functional mutations per individual sample). Based on an estimated number of approximately 30,000 mouse genes, the parallel sperm/DNA archive should permit the identification and recovery of ten or more alleles per average target gene which translates to a calculated 99% success rate in the discovery of five allelic variants for any given average gene. The low rate of unrelated ENU-induced passenger mutations has no practical impact on the analysis of the allele-specific phenotype at the G3 generation because of dilution and free segregation of such unrelated passenger mutations. To date 39 mouse models representing 33 different genes have been recovered from our archive using in vitro fertilization techniques. The generation time for a murine model heterozygous for a mutation in a gene of interest is less than 2 months, i.e., three to four times faster compared with current embryonic stem-cell-based technologies. We conclude that ENU-based targeted mutagenesis is a powerful tool for the fast and high-throughput production of murine gene-specific models for biomedical research.

[Solid-pseudopapillary tumors in childhood]. / Solid-pseudopapillärer Pankreastumor im Kindesalter, Ein Bericht über 5 Fälle, Germany.

We report on fife female patients with solid pseudopapillary tumors of the pancreas. The tumors are extremely rare in children. They occur mainly in adolescent and young adult females. The tumors are neoplasms of low malignancy with infrequent metastases, for instance, in the liver or the peritoneum. Although the tumors had reached a large diameter, all of them underwent complete tumor resection. After that the patients have a very good prognosis. Thus, so it is important to distinguish solid-pseudopapillary tumors from other tumors of the pancreas.

Intravenous C-type natriuretic peptide augments behavioral and endocrine effects of cholecystokinin tetrapeptide in healthy men.

Given the anxiogenic effects of the type-B natriuretic peptide receptor agonist C-type natriuretic peptide (CNP) in rodents, we investigated the influence of CNP pretreatment upon the behavioral and endocrine action of the panicogen cholecystokinin tetrapeptide (CCK-4) in healthy men. In a randomized double-blind balanced design, 20 male volunteers were given an intravenous infusion of 300 microg of CNP vs. placebo followed by 25 microg of CCK-4. The behavior was assessed using panic, anxiety, and dissociation questionaires before the infusion and after the CCK-4 stimulus. Furthermore, the stress-sensitive hormones adrenocorticotropic hormone (ACTH), cortisol, and prolactin were measured. CNP pretreatment enhanced the anxiogenic and prodissociative effects of CCK-4 and significantly augmented the ACTH surge after CCK-4. However, no effect of CNP was seen upon panic symptoms. Our preliminary data support a role of type-B natriuretic peptide receptors in anxiety modulation in normal man.

Anxiolytic activity of atrial natriuretic peptide in patients with panic disorder.

OBJECTIVE: Preclinical evidence exists for the anxiolytic activity of atrial natriuretic peptide, which is released during lactate-induced panic attacks. Atrial natriuretic peptide receptor modulation may have antipanic activity in patients with panic disorder. METHOD: The effects of 150 microg of atrial natriuretic peptide and placebo on panic attacks induced by cholecystokinin tetrapeptide (CCK-4) (25 microg) were studied in 10 panic disorder patients. The panicogenic activity of CCK-4 was measured with the Acute Panic Inventory. RESULTS: Panic attacks occurred in seven patients in the placebo condition and in two patients in the atrial natriuretic peptide condition. CCK-4 administration was accompanied by a significant increase in Acute Panic Inventory scores. Pretreatment with atrial natriuretic peptide resulted in significantly lower Acute Panic Inventory scores than pretreatment with placebo. CONCLUSIONS: The results support the antipanic activity of atrial natriuretic peptide. Nonpeptidergic atrial natriuretic peptide receptor ligands may be ultimately used to treat anxiety disorders.