Evaluation of the duration of human factor VIII expression in nonhuman primates after systemic delivery of an adenoviral vector

An E1/E2a/E3-deficient adenoviral vector encoding an epitope-tagged (flagged) human factor VIII (FVIII) cDNA was delivered systemically to four cynomolgus monkeys. Analysis of liver biopsy samples revealed the presence of vector DNA at all points in the study (day 7, 28, and 56), with vector copy number declining approximately 10-fold between day 7 and day 56. Immunoprecipitation/Western analyses detected human flagged FVIII in the plasma of all monkeys and expression persisted for 14-28 days. Peak plasma FVIII levels ranged from 50 to 100 ng/ml. Bethesda assays revealed no inhibitor in two animals, the development of a low-level transient inhibitor in one animal, and an inhibitor titer that continued to increase for the duration of the study in one animal. Other treatment-related changes included modest increases in liver enzymes, an increase in interleukin-6 (IL-6) levels, and a transient decrease in platelets in all four animals. These data indicate that early generation adenoviral vectors do not support the long-term expression of FVIII in nonhuman primates.     

Lipid microdomains are involved in neurospecific binding and internalisation of clostridial neurotoxins

The neuroparalytic syndromes of tetanus and botulism are caused by tetanus and botulinum neurotoxins, which are produced by bacteria of the genus Clostridia. These neurotoxins are structurally organised in three-domains endowed with different functions: specific interaction with the neuronal surface, membrane translocation and specific cleavage of three key components of the neurotransmitter release apparatus. Despite an identical intracellular activity, tetanus and botulinum neurotoxins are characterised by a differential intraneuronal trafficking, which is likely to be responsible for the different symptoms observed in clinical tetanus and botulism. This review aims to highlight recent discoveries on the recruitment of clostridial neurotoxins (CNTs) to the surface of neurons and neuronally-differentiated cell lines and to discuss their relevance for the internalisation and sorting of these neurotoxins.     

Esophageal implantation metastasis from adenocarcinoma of the cardia

A 74 years old male patient was admitted to our department suffering from dysphagia for five months. Gastric Barium studies showed a cardia tumour with polypoid lesions in the wall of the esophagus, and gastro-esophageal reflux disease was also diagnosed. Endoscopy verified a cardia tumour with esophageal metastasis, and biopsy was obtained from the two lesions. Histology showed that both tumours were adenocarcinomas. Endoscopic ultrasonography classified the cardia tumour as grade T2. As observed during endoscopy, the wall of the esophagus at the level of the polypoid lesion was hypo-echogenic and thick which was result of thickened mucosa. Total gastrectomy and oesophagectomy was performed. Pathology showed that the cardia tumour was pT2N2 and type Siewert-Stein II. The esophageal polypoid lesion was also proved an adenocarcinoma, which was localized only to the mucosa. No tumour cells were found in the blood- or in lymph vessels between the tumour and the esophageal adenocarcinoma. We think that the polypoid adenocarcinoma in the esophagus is an esophageal metastasis implantation from the cardia adenocarcinoma. It is based on the exclusion of other possible tumour dissemination routes. Probably the gastro-esophageal reflux is responsible for the implantation of tumour cells.     

Erythropoietin treatment in metastatic breast cancer--effects on Hb,quality of life and need for transfusion

Erythropoietin is an effective treatment for anemia in patients with various types of cancers, but few studies have evaluated the benefit of treatment in advanced breast cancer. In this multicenter study, we investigated the influence of two different doses of epoetin-beta on the level of hemoglobin, the need for blood transfusion, quality of life and safety aspects in patients with metastatic breast cancer. A total of 180 patients were randomized to receive either 1000 IE or 5000 IE epoetin-beta subcutaneously three times per week for 24 weeks. An increase of 20 g/L was defined as a positive hemoglobin response. Blood transfusions were given, if clinically indicated. Additional laboratory values and adverse events were recorded. Quality of life was measured with the aid of the EORTC QLQ-C30 questionnaire. Hemoglobin levels increased significantly in both groups. In the high-dose group, the initial mean Hb value was 98 g/L (64-110), which increased to 121 g/L (83-165) by week 24. In the low-dose group, the mean Hb value was 99 g/L (77-110.5) and by week 24 it was 116 g/L (81-144). The majority of patients who responded to treatment did so during the first four weeks. After 4 weeks, 7 patients in the low-dose group and 24 patients in the high-dose group had increased their Hb values by more than 20 g/L. The need for transfusion was low and did not differ between the groups. Quality of life was significantly enhanced in both groups, and there was no difference in the global quality of life between the two study arms. Epoetin-beta is a well-tolerated, safe and effective treatment of anemia in patients with metastatic breast cancer. There were significant improvements in Hb levels and quality of life in both groups.     

Subcortical neglect: Neuropsychological,SPECT, and neuropathological correlations with anterior choroidal artery territory infarction

In 2 patients with infarction in the territory of the right anterior choroidal artery, hemiparesis, hemihypesthesia (in 1), and hemianopia or superior quadrantanopia were associated with severe multimodal hemineglect, without anosognosia, disorientation, or asomatognosia. Single-photon emission tomography showed that marked hypoperfusion was not limited to the right posterior capsular region, but also involved the overlying parietal cortex, and to a lesser extent the frontal cortex. At autopsy in 1 patient, the infarct was nearly limited to the deep white matter of the temporal isthmus and the retrolenticular part of the internal capsule; only minute lesions were present in the globus pallidus, body of caudate, and amygdala. These findings are consistent with a disconnection phenomenon as the basis for subcortical neglect with ipsilateral deactivation of the parietofrontal cortex.     

Cefazolin prophylaxis in neurosurgery monitored by capillary electrophoresis

Prophylactic use of antibiotics to prevent postoperative infections is a routine method in neurosurgery. Little is known about the period of effectiveness of antibiotics applied only for the purposes of operation. The actual concentration of cefazolin was determined in the serum, in the contents of wound drains, and in the cerebrospinal fluid in a 24-hour postoperative period after the administration of 1 g of cefazolin just prior to skin incision in 8 patients undergoing lumbar discectomy and 11 patients undergoing craniectomy. The concentration of the antibiotic was then compared with the minimal inhibitory concentration values of cefazolin for 10 different bacterial species. For evaluating the concentration of cefazolin, capillary electrophoresis was used, which is a new clinical application of this separation technique. Results showed that the antibiotic was effective against bacterial breeding in the serum and in the drainage up to 12 hours. The drug concentration in the cerebrospinal fluid remained less than the value of the serum, and it exceeded the minimal inhibitory concentration values only for approximately 5 hours.     

Ulnar/fibular ray defect and brachydactyly in a family: a possible new autosomal dominant syndrome

The ulnar-mammary syndrome (MIM 181450) includes postaxial ray defects, abnormalities of growth, delayed sexual development, and mammary and apocrine gland hypoplasia. Brachydactyly type E (MIM 113300) presents with shortening of the metacarpals and phalanges in the ulnar ray in association with moderately short stature. We describe a three-generation family with variable expression of ulnar/fibular hypoplasia, brachydactyly, ulnar ray defects and short stature. The proband had ulnar hypoplasia with missing IV-Vth fingers, fibular hypoplasia on the right, bilateral club feet, growth retardation, a hypoplastic mid-face, an ASD and hemangiomas. She had normal mammary tissue and normal sweating. The mother had short stature, midfacial hypoplasia, a hypoplastic ulna and hypoplasia of the IVth metacarpal (brachydactyly) on the right without other associated malformations. The maternal grandfather had mild bilateral fibular hypoplasia and midphalangeal brachydactyly of the IV-Vth toes. His sister had mild short stature and shortening of the IVth metacarpal of the left hand. Two-point linkage analysis with microsatellite markers spanning the Ulnar-Mammary locus at 12q24.1 did not confirm linkage. The patients may have a previously undescribed syndrome.