Extending the honey bee venome with the antimicrobial peptide apidaecin and a protein resembling wasp antigen 5.

Honey bee venom is a complex mixture of toxic proteins and peptides. In the present study we tried to extend our knowledge of the venom composition using two different approaches. First, worker venom was analysed by liquid chromatography-mass spectrometry and this revealed the antimicrobial peptide apidaecin for the first time in such samples. Its expression in the venom gland was confirmed by reverse transcription PCR and by a peptidomic analysis of the venom apparatus tissue. Second, genome mining revealed a list of proteins with resemblance to known insect allergens or venom toxins, one of which showed homology to proteins of the antigen 5 (Ag5)/Sol i 3 cluster. It was demonstrated that the honey bee Ag5-like gene is expressed by venom gland tissue of winter bees but not of summer bees. Besides this seasonal variation, it shows an interesting spatial expression pattern with additional production in the hypopharyngeal glands, the brains and the midgut. Finally, our immunoblot study revealed that both synthetic apidaecin and the Ag5-like recombinant from bacteria evoke no humoral activity in beekeepers. Also, no IgG4-based cross-reactivity was detected between the honey bee Ag5-like protein and its yellow jacket paralogue Ves v 5.

Correlation of histology of healed fractures and tissue surrounding implants with ultrasonographic and radiographic appearance.

OBJECTIVES: To determine if there was histological correlation with ultrasonographic images of healed fractures and implant-associated tissue after fracture treatment by plate osteotomy. METHODS: Eight adult dogs were included in this retrospective study. Ultrasonography (B-mode and power Doppler) and radiography were performed before plate removal. Surgical biopsies were taken of the fracture site and the tissue adjacent to the plate. These were stained with haematoxylin and eosin to assess histomorphology and bone content and immunolabelled with CD31 to assess vascularity. RESULTS: Ultrasound and radiographic diagnoses of a healed fracture correlated with histological finding of bone healing. Ultrasonography and histology findings of vascularity were also highly correlated. The tissue surrounding the surgical implants was significantly more vascularised on both ultrasonography and histology than that at the fracture site itself. CLINICAL RELEVANCE: Ultrasonography can be used to diagnose fracture healing in plated fractures. Power Doppler ultrasonography examination of fracture healing should be performed away from surgical implants to avoid false-positive results of vascularisation.

Ultrasonographic assessment of fracture healing after plate osteosynthesis.

The goals of this study were to assess the ability of ultrasonography (US) to assess uncomplicated fracture healing and to establish normal images. Twenty-two dogs, ranging in age from 6 to 180 months were studied. Body weight ranged from 2.2 to 60 kg. All fractures were treated by plate osteosynthesis. US (B-mode and power Doppler) and radiography were performed until both were consistent with complete healing. B-mode US was performed in all dogs, and power Doppler US in 14. Fracture healing was judged to be complete based on US earlier than when based on radiography. The tissue immediately adjacent to the plate appeared vascularized on power Doppler images at a time when the tissue at the fracture site had a negative power Doppler exam. US appears useful for assessment of primary fracture healing and power Doppler was useful for detecting vascularization at the fracture site in nonhealed fractures. Power Doppler interrogation should be performed away from any metal implant, as a result from an interrogation adjacent to an implant will not reflect actual vascularization at the fracture site.

The use of an external skeletal traction device for distal fractures in the dog. A clinical case series of 11 patients.

Metacarpal and metatarsal fractures in 11 patients were treated 'closed' with a specially designed External Skeletal Fixation (ESF) frame with a walking bar and traction applied to the digits. Mediolateral angulation had improved postoperatively in 10 of the 11 patients. Craniocaudal angulation had improved in eight patients and could not be evaluated in three due to ESF frame superimposition. At follow-up, eight patients had a good clinical function, whereas three patients were still slightly lame. Only minor ESF-related complications were seen (pin loosening, pin tract infection, cerclage wire breakage and bending of the frame), which resolved without intervention after frame removal.

Off-pump coronary artery grafting in patients necessitating multiple distal anastomoses.

OBJECTIVES: Off-pump surgery has become a valuable alternative if a small number of distal anastomoses is required. The aim of the present study was to test the hypothesis that outcome is not altered in multiple distal anastomoses. MATERIAL AND METHODS: During a 4 year period, 350 patients were operated off-pump. 187 patients received one or two distal anastomoses (group A), and 163 patients received more than two distal anastomoses (group B). Pre-operative characteristics, intra-operative details and postoperative outcome were reviewed. RESULTS: In group A, 1.8 +/- 0.4 distal anastomoses were performed, versus 3.2 +/- 0.5 in group B (P < 0.001). There were more sequential distal anastomoses in group B (1.09 in group A versus 1.38 in group B) with a wider use of right internal thoracic and radial arteries. There were more anastomoses performed on the lateral (89% vs. 45%) and on the inferior wall (65% vs. 16%) in group B, compared with group A. The operation time was significantly longer (206 +/- 75 minutes versus 158 +/- 33 minutes) and the operative blood loss significantly higher (748 +/- 516 ml versus 509 +/- 361 ml) in group B. Total blood loss, transfusion requirements, neurological disfunction, hospital stay, troponine I level, postoperative infarction and 30 days mortality did not differ significantly between groups. There were 11 conversions (3.1%), 3 patients in group A and 8 patients in group B (NS). Most conversion occurred imperative while performing an anastomosis on the anterior wall. CONCLUSION: Beating heart surgery with multiple distal anastomoses, can be performed without increased myocardial damage, postoperative morbidity or hospital mortality. Lateral wall grafting could not be identified as a trigger for conversion.

MR imaging of the pelvic floor.

Pelvic floor disorders are a common clinical problem. Clinical evaluation frequently underestimates the extent of the disease. Fluoroscopic colpocystodefecography (CCD) is an established tool for diagnosing disorders of the rectum and recto- anal junction. It is however less performant when it comes to evaluate the complex pelvic floor mechanism in all its aspects. MR defecography allows to evaluate all the compartments of the pelvis without being invasive. This article describes the clinical setting, the MR technique and provides guidelines for interpretation of a MR defecography study.

The effect of botulinum-A toxin in incontinent children with therapy resistant overactive detrusor.

PURPOSE: We determined the effect of detrusor injection of botulinum-A toxin in a cohort of children with therapy resistant non-neurogenic detrusor overactivity. This prospective study included therapy resistant children with overactive bladder. MATERIAL AND METHODS: During the study period of 19 months 10 boys and 11 girls were included. All patients showed decreased bladder capacity for age, urge and urge incontinence. Main treatment duration before inclusion was 45 months. A dose of 100 U botulinum-A toxin (Botox) was injected in the detrusor. RESULTS: Side effects were evaluated in all 21 included patients. The side effects reported were 10-day temporary urinary retention in 1 girl and signs of vesicoureteral reflux with flank pain during voiding in 1 boy, which disappeared spontaneously after 2 weeks. No further examinations were done since the boy refused. Two girls experienced 1 episode each of symptomatic lower urinary tract infection. Eight girls and 7 boys with a minimum followup of 6 months represent the study group for long-term evaluation. In this study group after 1 injection 9 patients showed full response (no more urge and dry during the day) with a mean increase in bladder capacity from 167 to 271 ml (p <0.001). Three patients showed a partial response (50% decrease in urge and incontinence) and 3 remained unchanged. Eight of the 9 full responders were still cured after 12 months, while 1 of the initially successfully treated patients had relapse after 8 months. The 3 partial responders and the patient with relapse underwent a second injection with a full response in the former full responder and in 1 partial responder. CONCLUSIONS: Botulinum-A toxin injection in children with non-neurogenic overactive detrusor is an excellent treatment adjunct, leading to long-term results in 70% after 1 injection.

Cystectomy and orthotopic ileal neobladder construction. Evaluation of continence and complications in a regional hospital.

INTRODUCTION: This study evaluates the outcome of orthotopic neobladder construction in a regional hospital setting. MATERIAL AND METHODS: Orthotopic ileal neobladders were constructed in 51 patients between 1990 and 2001. All complications were reported systematically and continence was evaluated in a standardized manner. RESULTS: One patient died perioperatively (2%). Early complications occurred in 56.9%. They were related to the neobladder in 21.6% and unrelated to the neobladder in 47.1%. 31.4% experienced complications in the late postoperative period. In 25.5%, these complications were neobladder related and in 11.8% they were not. After 1 year, daytime continence was 91.7% and nighttime continence 80.6%. Our results are comparable to those reported by others. CONCLUSION: We conclude that the construction of a Hautmann neobladder can be performed with acceptable results in a regional hospital. The democratization of recent bladder replacement techniques may offer more patients a better quality of life and prevent the formation of waiting lists. Radical treatment may then be opted for sooner and thus improve cancer-related survival.

Peptide profiling of a single Locusta migratoria corpus cardiacum by nano-LC tandem mass spectrometry.

The pars intercerebralis-corpora cardiaca complex in insects is the functional equivalent of the vertebrate brain-pituitary axis. During the past few decades more than 40 neuropeptides have been isolated from the locust brain-corpus cardiacum complex. Tedious and time-consuming successive purification rounds of large tissue extracts were necessary to achieve the purification and sequencing of most of these signal molecules. Nowadays, the combination of nanoscale liquid chromatography and the very sensitive tandem mass spectrometry allows us to identify and sequence peptides in very low concentration directly from tissue extracts. In this manuscript, we review previous data on the peptidome analysis of the locust corpora cardiaca, with emphasis on AKH processing. In addition, we report the peptide profiling of a single corpus cardiacum from Locusta migratoria. 23 peptides were isolated and sequenced in a single nano-LC-MS/MS experiment, demonstrating the sensitivity and effectiveness of mass spectrometry in peptide research.

Effect of Bacillus thuringiensis Cry1 toxins in insect hemolymph and their neurotoxicity in brain cells of Lymantria dispar.

Little information is available on the systemic effects of Bacillus thuringiensis toxins in the hemocoel of insects. In order to test whether B. thuringiensis-activated toxins elicit a toxic response in the hemocoel, we measured the effect of intrahemocoelic injections of several Cry1 toxins on the food intake, growth, and survival of Lymantria dispar (Lepidoptera) and Neobellieria bullata (Diptera) larvae. Injection of Cry1C was highly toxic to the Lymantria larvae and resulted in the complete inhibition of food intake, growth arrest, and death in a dose-dependent manner. Cry1Aa and Cry1Ab (5 microg/0.2 g [fresh weight] [g fresh wt]) also affected growth and food intake but were less toxic than Cry1C (0.5 microg/0.2 g fresh wt). Cry1E and Cry1Ac (5 microg/0.2 g fresh wt) had no toxic effect upon injection. Cry1C was also highly toxic to N. bullata larvae upon injection. Injection of 5 microg/0.2 g fresh wt resulted in rapid paralysis, followed by hemocytic melanization and death. Lower concentrations delayed pupariation or gave rise to malformation of the puparium. Finally, Cry1C was toxic to brain cells of Lymantria in vitro. The addition of Cry1C (20 microg/ml) to primary cultures of Lymantria brain cells resulted in rapid lysis of the cultured neurons.

Clinical application of the Bardex IC Foley catheter.

OBJECTIVES: We performed two randomized prospective studies with the silver-coated Bardex IC catheter in order to evaluate the incidence of bacteriuria during short- and medium-term catheterization after urological procedures. METHODS: During catheterization only consecutive suprapubic urine samples were taken and cultured. After removal of the catheter the patient was allowed one wash-out void, and before the second micturition a suprapubic puncture was performed to collect a culture specimen. RESULTS: In the first trial, after radical prostatectomy 18 patients with the Bardex IC catheter were compared to 17 patients with a silicon catheter after the same procedure. There was no significant difference in bacteriuria after 14 days (50.0 vs. 53.3%). In the second part of the study 180 patients were evaluated 101 with latex and 79 with Bardex IC catheters. The median catheterization time was 5 days. The results show a significant delay in the onset of bacteriuria when a silver alloy catheter is used (p < 0.003). On day 5 only 6.3% had bacteriuria in the Bardex IC group versus 11.9% in the latex group. CONCLUSION: We conclude that, after urological procedures, short-term catheterization with the Bardex IC catheter is superior to the classical latex catheter.

In-vivo diagnosis of right renal aplasia in a pekingese bitch.

The physical, haematological, urinary and radiographic findings in a female pekingese dog with right renal aplasia are described. The right ureter was present and structurally normal. The left kidney was hypertrophied. The ovaries and uterus were normal.