Bacterial ghosts (BGs)—Advanced antigen and drug delivery system

Bacterial ghosts (BGs) are empty bacterial envelopes of Gram-negative bacteria produced by controlled expression of cloned gene E, forming a lysis tunnel structure within the envelope of the living bacteria. BGs are devoid of cytoplasmic content and possess all bacterial bio-adhesive surface properties in their original state while not posing any infectious threat. BGs are ideally suited as an advanced drug delivery system (ADDS) for toxic substances in tumor therapy. The inner space of BGs can be loaded with either single components or combinations of peptides, drugs or DNA which provides an opportunity to design new types of (polyvalent) drug delivery vehicles. Uptake of BGs loaded with Doxorubicin (Dox) by CaCo2 cells led to effective Dox release from endo-lysosomal compartments and accumulation in the nucleus. Viability and proliferative capacity of the cells were significantly decreased (2–3 orders of magnitude) after internalization of Dox loaded BGs as compared to cells incubated with free Dox. The same effect was observed with leukemia cells. Melanoma cells also revealed a high capability to internalize BGs. These results indicate that BGs are able to target a range of types of cancer. BGs have also been investigated as DNA delivery vectors. Studies show DNA loaded BGs are efficiently phagocytosed and internalized by both professional APCs and tumor cells with up to 82% of cells expressing the plasmid-encoded reporter gene. Our studies with BGs as an ADDS system contribute (i) to optimize drug delivery for the treatment of cancer; (ii) define specific conditions for selection and preparation of BG formulations; (iii) and provide a background for the clinical application of BGs in cancer therapy.     

Recombinant and nonrecombinant factor XIII and its effect on bone ingrowth and strength of fixation

Thirty cylindrical, commercially pure, titanium fiber, porous-coated Ti₆Al₄V implants were inserted pressfit into the proximal humeral portion of 30 sheep humeri to determine the systemic effect of recombinant factor XIII and placenta-derived factor XIII concentrate on bone ingrowth and strength of fixation. For both the recombinant factor XIII and the factor XIII concentrate group, the volume of bone ingrowth and the strength of fixation were higher than for the control specimens. However, the difference was only significant for the factor XIII concentrate group.     

Endoscopic sealing of infected bronchus stump fistulae with fibrin following lung resections

Summary Bronchus stump insufficiency following lung resection, with an average incidence of 4%, is a serious complication which carries a mortality of up to 90%. Operative transthoracic approaches have been largely unsatisfactory because of the high operative risk and rapidly spreading infection. In an experimental study on 18 pigs, endoscopic occlusion of infected bronchus stump fistulae was achieved with fibrin sealant (1 ml, 500 units/ml thrombin, 3500 units/ml aprotinin) applied via a flexible bronchoscope. During autopsy, all bronchus stump fistulae were found to have healed after the second postoperative week. Transitory local abscesses of the pleura could be prevented by high-dose systemic antibiotic therapy for 5 postoperative days but not by antibiotics added to the fibrin sealant. This endoscopic method has already been performed successfully in 3 clinical cases; additional sclerotherapy with (2–3 ml Ethoxysclerol applied around the fistula orifice was carried out before fibrin sealing to stimulate fibrosis. Endoscopic controls demonstrated fistula closure by granulation tissue after 2 weeks. This procedure could become the method of choice for infected postoperative fistulae of the bronchus stump and should be attempted in any case before operative approaches are considered.     

Blood coagulation activation and fibrinolysis during a downhill marathon run.

Prolonged physical exercise is associated with multiple changes in blood hemostasis. Eccentric muscle activation induces microtrauma of skeletal muscles, inducing an inflammatory response. Since there is a link between inflammation and coagulation we speculated that downhill running strongly activates the coagulation system. Thirteen volunteers participated in the Tyrolean Speed Marathon (42,195 m downhill race, 795 m vertical distance). Venous blood was collected 3 days (T1) and 3 h (T2) before the run, within 30 min after finishing (T3) and 1 day thereafter (T4). We measured the following key parameters: creatine kinase, myoglobin, thrombin-antithrombin complex, prothrombin fragment F1 + 2, D-dimer, plasmin-alpha(2)-antiplasmin complexes, tissue-type plasminogen activator antigen, plasminogen-activator-inhibitor-1 antigen and thrombelastography with ROTEM [intrinsic pathway (InTEM) clotting time, clot formation time, maximum clot firmness, alpha angle]. Thrombin generation was evaluated by the Thrombin Dynamic Test and the Technothrombin TGA test. Creatine kinase and myoglobin were elevated at T3 and further increased at T4. Thrombin-antithrombin complex, prothrombin fragment F1 + 2, D-dimer, plasmin-alpha(2)-antiplasmin complexes, tissue-type plasminogen activator antigen and plasminogen-activator-inhibitor-1 antigen were significantly increased at T3. ROTEM analysis exhibited a shortening of InTEM clotting time and clot formation time after the marathon, and an increase in InTEM maximum clot firmness and alpha angle. Changes in TGA were indicative for thrombin generation after the marathon. We demonstrated that a downhill marathon induces an activation of coagulation, as measured by specific parameters for coagulation, ROTEM and thrombin generation assays. These changes were paralleled by an activation of fibrinolysis indicating a preserved hemostatic balance.     

Endoscopically controlled erbium:YAG goniopuncture versus trabeculectomy: effect on intraocular pressure in combination with cataract surgery

PURPOSE:. To compare the efficacy of endoscopic erbium:YAG laser goniopuncture in glaucoma treatment to trabeculectomy, both methods as adjuncts to cataract surgery. METHODS:. Fifty-nine eyes of 59 glaucoma patients with coexistent cataract were treated by phacoemulsification and endoscopic Er:YAG goniopuncture in a combined fashion. The primary study endpoints were intraocular pressure (IOP), number of antiglaucomatous drugs, postoperative complications, hospitalisation time and visual acuity at 1 year after surgery. To date, 24 eyes have finished the 1-year follow-up. This prospective treatment arm was compared to a retrospective inclusion-matched control group treated by trabeculectomy and cataract surgery in a single procedure. RESULTS:. In the laser-treated group, the mean IOP dropped by 30% from 23.4+/-3.7 mmHg to 16.3+/-6 mmHg ( P<0.0001) after 12 months. Without reoperation, treatment was successful in 71% of these eyes. In the control group, the IOP decreased by 33.5% from 22.7+/-3.3 mmHg to 15.1+/-3.8 mmHg ( P<0.0001). The success rate without reoperation was 46%. The number of antiglaucomatous drugs needed decreased from 1.48+/-0.95 to 0.48+/-0.7 ( P<0.0001) in the laser-treated group and from 2.0+/-0.9 to 0.39+/-0.6 ( P<0.0001) in the control group. Postoperative complications were found more frequently in the control group ( P<0.0001). Hospitalisation was shorter in the laser group ( P<0.0001). Postoperative visual acuity was lower in the control group ( P=0.004). CONCLUSION:. Combined Er:YAG goniopuncture and cataract surgery lowers the IOP to an extent comparable to combined trabeculectomy and cataract surgery. Due to fewer postoperative complications, Er:YAG goniopuncture seems to be superior to standard fistulation surgery as the primary approach within the first year.     

Synthese neuer Indolmethanamine durch Leimgruber-Batcho Reaktion

Synthesis of New Indolemethanamines Using the Leimgruber-Batcho Reaction A short and efficient synthesis of the title compounds is reported starting with commercially available carboxylic acids 1 and 5 , respectively. By appropriate modifications in the course of the Leimgruber-Batcho reaction used in the key step quite stable 1-hydroxy-indole derivatives became available.     

Absence of insulin receptor gene mutations in three insulin-resistant women with the polycystic ovary syndrome

Women with polycystic ovary syndrome (PCOS) are markedly insulin-resistant, but the molecular mechanisms of these changes and their relationship to the hyperandrogenic state remain to be clarified. Mutations have recently been identified in the insulin receptor gene of patients with extreme forms of insulin resistance associated with hyperandrogenism (eg, type A insulin resistance), and these mutations account for the insulin resistance in such patients. We performed this study to determine whether mutations in the coding portion of the insulin receptor gene were responsible for insulin resistance in PCOS. Insulin binding studies using cultured skin fibroblasts of three obese (body mass index > 27 kg/m²) women with PCOS (ie, mild hyperandrogenemia and chronic anovulation of unknown etiology) and documented insulin resistance showed no apprarent abnormalities in either the number or affinity of insulin binding sites. Direct sequencing of all 22 exons of the insulin receptor gene from two of the women with PCOS did not reveal any mutations. Furthermore, both alleles of the gene were expressed at equal levels. In a third insulin-resistant PCOS woman, there was no evidence for a mutation in the coding portion of the insulin receptor gene as determined by denaturing gradient gel electrophoresis (DGGE). We conclude that the insulin resistance in these PCOS women was caused by a defect extrinsic to the insulin receptor.     

Cervical bruits: clinical correlates of stenosis

To analyze the relationship among characteristics of buits, degree of underlying stenosis, and neurologic symptom complexes, the authors studied 157 patients with bruits undergoing digital subtraction angiography. Symptom status assignment (definite lateralizing, possible lateralizing, vertebrobasilar, diffuse, and asymptomatic) and cervical auscultation for location, duration, and other bruit characteristics were performed independently. Bruit occurrence was associated with stenosis of greater than or equal to 50% of the underlying vessel with the association being no stronger at higher levels of stenosis (greater than or equal to 80%). No relationship existed between the side of bruit and side of symptoms in those with lateralized symptoms. The authors found more severe degrees of carotid stenosis in two symptomatic groups (vertebrobasilar insufficiency [VBI] and definite lateralized) compared with asymptomatic patients. Those with VBI had more high-grade stenosis, whereas those with lateralized symptoms had more occlusions, which tended to be ipsilateral to the symptoms.