siRNA mediated knockdown of tissue factor expression in pigs for xenotransplantation.

Acute vascular rejection (AVR), in particular microvascular thrombosis, is an important barrier to successful pig-to-primate xenotransplantation. Here, we report the generation of pigs with decreased tissue factor (TF) levels induced by small interfering (si)RNA-mediated gene silencing. Porcine fibroblasts were transfected with TF-targeting small hairpin (sh)RNA and used for somatic cell nuclear transfer. Offspring were analyzed for siRNA, TF mRNA and TF protein level. Functionality of TF downregulation was investigated by a whole blood clotting test and a flow chamber assay. TF siRNA was expressed in all twelve liveborn piglets. TF mRNA expression was reduced by 94.1 ± 4.7% in TF knockdown (TFkd) fibroblasts compared to wild-type (WT). TF protein expression in PAEC stimulated with 50 ng/mL TNF-α was significantly lower in TFkd pigs (mean fluorescence intensity TFkd: 7136 ± 136 vs. WT: 13 038 ± 1672). TF downregulation significantly increased clotting time (TFkd: 73.3 ± 8.8 min, WT: 45.8 ± 7.7 min, p < 0.0001) and significantly decreased thrombus formation compared to WT (mean thrombus coverage per viewing field in %; WT: 23.5 ± 13.0, TFkd: 2.6 ± 3.7, p < 0.0001). Our data show that a functional knockdown of TF is compatible with normal development and survival of pigs. TF knockdown could be a valuable component in the generation of multi-transgenic pigs for xenotransplantation.

Experimental comparison of low doses of 1.5 mg fluspirilene and bromazepam in out-patients with psychovegetative disturbances.

In order to find out whether the administration of bromazepam and low doses of fluspirilene for six weeks had different effects, 45 out-patients of both sexes received 6 mg bromazepam per day or 1.5 mg fluspirilene per week in a double-blind randomized fashion. Overall therapeutic effectiveness was rated by the physician in two weeks' intervals. In addition use was made of the Hamilton Anxiety Scale, the Adjective List (Janke and Debus), and the Symptom Check List (Derogatis). Both products reduced anxiety in a comparable manner. Under fluspirilene, however, global therapeutic improvement was often more obvious than under bromazepam. This therapeutic superiority was particularly remarkable in patients whose somatic concern in the meaning of the Hamilton Anxiety Scale was comparatively pronounced.

[Alternative therapy concept for the treatment of psychosomatic disorders. Controlled double-blind evaluation of fluspirilene versus bromazepam]. / Alternatives Therapiekonzept zur Behandlung psychosomatischer Beschwerden. Kontrollierte Doppelblind-Prüfung von Fluspirilen versus Bromazepam.

Today, Fluspirilene 1.5 mg i.m. (Imap 1,5 mg), is used to treat psychosomatic conditions, anxiety states and psychogenic disorders. The present article reports on clinical studies involving these indications. The efficacy of Fluspirilene 1.5 mg i.m. was demonstrated in patients with functional organic complaints, autonomic emotional symptom complexes, patients with anxiety, patients with functional heart complaints, and patients with functional gynaecological disorders. The results of a controlled study of 1.5 mg Fluspirilene i.m. and 6 mg Bromazepam are discussed. Forty-five patients from a neurologist's practice participated, all of whom had been classified as treatment-requiring patients with neurotic anxiety, on the basis of the physician's diagnosis and psychometric examination. Treatment lasted 42 days. The clinical assessment by the physician revealed that 1.5 mg Fluspirilene i.m. is significantly more effective than Bromazepam. The psychometric findings showed that patients with marked somatic anxiety treated with Fluspirilene showed greater improvement as expressed in a reduction in anxiety, lightening of mood, and increased activity, as compared with Bromazepam.