Single Nucleotide Primer Extension (SNuPE) analysis of the G6PD gene in somatic cells and oocytes of a kangaroo (Macropus robustus)

cDNA sequence analysis of the X-linked glucose-6-phosphate dehydrogenase (G6PD) gene has shown a base difference between two subspecies of the kangaroo, Macropus robustus robustus (wallaroo) and M. r. erubescens (euro). A thymine residue in the wallaroo at position 358 in exon 5 has been replaced by a cytosine residue in the euro, which accounts for the previously reported electrophoretic difference between the two subspecies. This base difference allowed use of the Single Nucleotide Primer Extension (SNuPE) technique to study allele-specific expression of G6PD at the transcriptional level. We began by examining G6PD expression in somatic cells and observed complete paternal X inactivation in all somatic tissues of adult female heterozygotes, whereas we found partial paternal allele activity in cultured fibroblasts, thus confirming previous allozyme electrophoresis studies. In late dictyate oocytes from an adult heterozygote, the assay also detected expression of both the maternal and paternal alleles at the G6PD locus, with the maternal allele showing preferential expression. Thus reactivation of the inactive paternally derived X chromosome occurs during oogenesis in M. robustus, although the exact timing of reactivation remains to be determined

Detection of bacterial biofilm in double capsule surrounding mammary implants : findings in human and porcine breast augmentation

The finding of a double capsule surrounding breast implants is a recognized complication of breast augmentation surgery. In a recent review, double capsules were identified in 14 of 626 breast implants. We report the detection of incidental double capsules both in a patient and from our previously described porcine model.3 page(s

Preoperative abdominal muscle elongation with botulinum toxin A for complex incisional ventral hernia repair

Background: Surgical repair of recurrent abdominal incisional hernia(s) can be challenging due to complex operative conditions, intense post-operative pain, potential respiratory compromise and lateral muscle traction predisposing to early recurrence. We report our preliminary results with botulinum toxin A (BTA) injection causing flaccid paralysis (relaxation) of the lateral abdominal wall muscles prior to surgery. Methods: A prospective pilot study measured the effect of preoperative BTA prior to elective repair of recurrent abdominal hernias. Under ultrasound control, 2 weeks prior to surgery, 50 units of BTA was injected into the external oblique, internal oblique and transversus abdominis muscles at three sites on each side of the lateral abdominal wall (total dose 300 units). Pre- and post-BTA abdominal computed tomography measured changes in abdominal wall muscle thickness and length. All hernias were repaired with laparoscopic or laparoscopic-assisted mesh techniques in a single or two-staged procedure. Results: Eight patients received BTA injections which were tolerated with no complications. Post-BTA preoperative computed tomography showed a significant increase in mean length of lateral abdominal wall from 18.5cm pre-BTA to 21.3cm post-BTA (P = 0.017) with a mean unstretched length gain of 2.8cm per side (range 0.8-6.0cm). All hernias were surgically reduced with mesh with no early recurrence. Conclusion: Preoperative BTA injection prior to complex abdominal hernia repair is a safe procedure that causes flaccid relaxation, elongation and thinning of the lateral abdominal muscles and decrease in hernia defect. Although further evaluation is required, BTA injections may be a useful adjunct to surgical repair of complex incisional hernias

Presence of biofilm containing viable multiresistant organisms despite terminal cleaning on clinical surfaces in an intensive care unit

Background: Despite recent attention to surface cleaning and hand hygiene programmes, multiresistant organisms (MROs) continue to be isolated from the hospital environment. Biofilms, consisting of bacteria embedded in exopolymeric substances (EPS) are difficult to remove due to their increased resistance to detergents and disinfectants, and periodically release free-swimming planktonic bacteria back into the environment which may may act as an infection source. Aim: To establish whether reservoirs of MROs exist in the environment as biofilms. Methods: Following terminal cleaning, equipment and furnishings were removed aseptically from an intensive care unit (ICU) and subjected to culture and scanning electron microscopy (SEM). Samples were placed in 5 mL of tryptone soya broth, sonicated for 5 min before plate culture on horse blood agar, Brillance MRSA and Brilliance VRE agar plates. Samples for SEM were fixed in 3% glutaraldehyde and hexamethyldisilizane (HMDS) prior to sputter-coating with gold and examination in an electron microscope. Findings: Biofilm was demonstrated visually on the sterile supply bucket, the opaque plastic door, the venetian blind cord, and the sink rubber, whereas EPS alone was seen on the curtain. Viable bacteria were grown from three samples, including MRSA from the venetian blind cord and the curtain. Conclusion: Biofilm containing MROs persist on clinical surfaces from an ICU despite terminal cleaning, suggesting that current cleaning practices are inadequate to control biofilm development. The presence of MROs being protected within these biofilms may be the mechanism by which MROs persist within the hospital environment

Minimally invasive Venetian blinds ventral hernia repair with botulinum toxin chemical component separation

Background: Laparoscopic ventral repair is safe, with lower wound infection rates compared with open repair. ‘Venetian blinds’ technique of plication in combination with mesh reinforcement, is totally intra‐corporeal, with hernia defect and sac plication to reduce seroma formation. While laparoscopic suturing of the abdominal wall can represent a technical challenge, pre‐operative botulinum toxin A (BTA) injections as an adjunct can assist. This study aims to demonstrate feasibility and efficacy of this technique in abdominal wall hernia repair, with BTA adjunct in midline hernias. Methods: A single‐centre case series was conducted using minimally invasive ‘Venetian blinds’ technique for repair of complex ventral abdominal hernias. Twelve patients (seven midline, five non‐midline) underwent repair (11 laparoscopic; one robotic). Midline hernias received BTA (200–300 units Botox) 4–6 weeks prior to surgery. Repairs were mesh‐reinforced following fascial closure. Results: Twelve (10 female, two male) patients, with a median age 72 years (range 31–83) and body mass index of 27.3 kg/m2 (range 22.8–61.7) were included. The median length of operation was 133 min (range 45–290) and length of hospital stay 3 days (range 1–28). To date there has been no recurrence of hernia. A single symptomatic seroma was treated with antibiotics and did not require mesh removal. One patient developed hospital‐acquired pneumonia and pseudomembranous colitis. Conclusion: Minimally invasive ‘Venetian blinds’ technique has promising early results with both midline and non‐midline ventral hernias. The addition of BTA is a novel and feasible combination for repair of midline ventral hernias

Preoperative chemical component relaxation using Botulinum toxin A : enabling laparoscopic repair of complex ventral hernia

Background: Repair of complex ventral hernia can be very challenging for surgeons. Closure of large defects can have serious pathophysiological consequences. Botulinum toxin A (BTA) has recently been described to provide flaccid paralysis to abdominal muscles prior to surgery, facilitating closure and repair. Methods: This was a prospective observational study of 32 patients who underwent ultrasound-guided injections of BTA to the lateral abdominal wall muscles prior to elective repair of complex ventral hernia between January 2013 and December 2015. Serial non-contrast abdominal CT imaging was performed to measure changes in fascial defect size, abdominal wall muscle length and thickness. All hernias were repaired laparoscopically or laparoscopic-assisted with placement of intra-peritoneal mesh. Results: Thirty-two patients received BTA injections which were well tolerated with no complications. A comparison of baseline (preBTA) CT imaging with postBTA imaging demonstrated an increase in mean baseline abdominal wall length from 16.4 to 20.4 cm per side (p < 0.0001), which translates to a gain in mean transverse length of the unstretched anterolateral abdominal wall muscles of 4.0 cm/side (range 0–11.7 cm/side). Fascial closure was achieved in all cases, with no instances of raised intra-abdominal pressures or its sequelae, and there have been no hernia recurrences to date. Conclusions: Preoperative BTA injection to the muscles of the anterolateral abdominal wall is a safe and effective technique for the preoperative preparation of patients prior to laparoscopic mesh repair of complex ventral hernia. This technique elongates and thins the contracted and retracted musculature, enabling closure of large defects

Laparoscopic repair of complex ventral hernia facilitated by pre-operative chemical component relaxation using Botulinum Toxin A

Purpose: The operative management of complex ventral hernia poses a formidable challenge, despite recent advances in surgical techniques. Recurrence rates after complex ventral hernia repair remain high, and increase with each failed attempt. This study examines the effect of pre-operative abdominal wall chemical component relaxation using Botulinum Toxin A (BTA) to induce temporary flaccid paralysis in order to facilitate laparoscopic repair of large complex ventral hernia. Methods: This is a prospective evaluation of 27 patients from January 2013 to August 2015 who underwent ultrasound guided BTA injections to the lateral abdominal wall muscles prior to elective complex ventral hernia repair. Non-contrast serial CT imaging was obtained pre- and post-BTA injection to measure change in fascial defect size and abdominal wall muscle thickness and length. Fascial defects were closed and hernias repaired using laparoscopic or laparoscopic-assisted intra-peritoneal onlay mesh (IPOM) techniques. Results: 27 patients received pre-operative BTA injections which were well tolerated with no complications. Comparison of pre-BTA and post-BTA CT imaging demonstrated a significant increase in mean length of the lateral abdominal wall from 15.7 cm pre-BTA to 19.9 cm post-BTA (p < 0.0001), with mean unstretched length gain of 4.2 cm/side (range 0–11.7 cm/side). All hernias were surgically reduced and repaired with mesh, with no early recurrences. Conclusion: Pre-operative administration of BTA is a safe and effective technique in the pre-operative preparation of patients undergoing elective complex ventral hernia repair. This technique lengthens and relaxes the laterally retracted abdominal muscles and enables laparoscopic closure of large complex ventral hernia