Chronic cranial window implantation for high-resolution intravital imaging of the endothelial glycocalyx in mouse cortex.

The endothelial glycocalyx is an integral component of the brain vascular barrier. Visualizing its structure in vivo is essential to understand its physiological and pathophysiological mechanisms. Here, we present a surgical protocol for chronic cranial window implantation in mice, alongside the use of multiphoton microscopy tools to image the cortical vasculature. We describe steps for cranial window implantation, intravenous injection of fluorescent markers, and intravital imaging. We then detail a technique to quantify glycocalyx thickness using Imaris image analysis software. For complete details on the use and execution of this protocol, please refer to Gray et al. (2023).1.

Radiotherapy for anal squamous cell carcinoma: must the upper pelvic nodes and the inguinal nodes be treated?

BACKGROUND: Loco-regional failure is the predominant cause of death in anal squamous cell carcinoma. We assessed patterns of loco-regional recurrence to determine the impact of radiotherapy (RT) volumes on patient outcome. METHODS: Retrospective clinical study, including patients treated curatively with RT or chemo-radiotherapy between 1994 and 2007. RT fields/volumes were reviewed and compared with patterns of failure. Patients were classified as having whole pelvic radiotherapy (WPRT) if RT extended to L5/S1 or lower pelvic radiotherapy (LPRT) if it extended to the lower sacroiliac joints or below. Patients with negative inguinal nodes either underwent prophylactic inguinal radiotherapy (PIRT) or had inguinal observation (IO). Patterns of failure were compared. RESULTS: Twenty-seven patients (53%) had WPRT and 24 (47%) had LPRT. Forty-two patients had negative inguinal nodes: 29 (69%) had PIRT and 13 (31%) had IO. Median follow-up was 5.8 years. Twelve regional failures occurred in eight patients: three pelvic, one inguinal and four pelvic and inguinal. All patients with regional failure died of disease. Pelvic nodal failure was 7.7% in N0 and 33% in N1-3 patients (P = 0.012). There was no difference in pelvic regional failure between WPRT and LPRT (11% versus 16%, P = 0.64). There was only one possible regional failure above LPRT in this group (4%). Inguinal failure was 0% in the PIRT group compared with 23% in IO group (P = 0.009). CONCLUSION: There was no difference in pelvic regional failure between WPRT and LPRT. LPRT is likely to be safe in N0 patients. Inguinal nodes should be treated in all patients.

Community health indicators associated with outcomes of pancreatectomy.

PURPOSE: This study evaluates the association of environmental, social and health risk factors in relation to outcomes of pancreatic surgery. METHODS: Patients who underwent pancreatectomy with a 30 day postoperative follow up in Florida, New York and Washington states were identified using the State Inpatient Databases (SID) from 2010 to 2011. This data was merged with community health indicators complied from the County Health Ranking database. Fourteen community health indicators were used to determine higher risk communities. Communities were then divided into low and high risk communities based on a scoring system using accumulative community risk. RESULTS: Among 3494 patients included recipients in high-risk communities were more likely African American (p < 0.001), younger (age 40-59; p = 0.001), and had Medicaid as primary insurance (p = 0.001). Management of patients in high-risk communities was associated with increased risk of postoperative complications (p < 0.001), ICU admissions (p < 0.001), increased length of stay (p < 0.001). CONCLUSION: Health indicators from patients' communities are predictors of increased risk of perioperative complications for individuals undergoing pancreas surgery.

Achieving quality in colonoscopy: bowel preparation timing and colon cleanliness.

BACKGROUND: Colonoscopy is considered the gold standard for investigation of large bowel pathology. Numerous factors influence the efficacy of bowel preparation for colonoscopy. Inadequate bowel preparation can lead to missed pathology. Timing of fasting and bowel preparation, timing of procedure and possibly patient bowel habit and presence of diverticula may have an influence on the quality of the preparation. The aim of this study was to investigate the quality of cleansing of sodium picosulfate (Picoprep-3™, Pharmatel Fresenius Kabi Pty Ltd, Pymble, NSW, Australia) with different administration schedules and to evaluate whether patient's bowel patterns influence the quality of cleansing. METHODS: Three hundred twenty-five patients (175 morning and 150 afternoon procedures) were interviewed prior to colonoscopy to evaluate bowel habit and timing of preparation administration. Quality of cleansing was then assessed during colonoscopy using a 5-point scale. Further factors analysed included the patient's prior bowel habit and the presence of diverticula at colonoscopy. Procedural end points evaluated included procedure total time, caecal intubation time and withdrawal times. RESULTS: The quality of cleansing for individual bowel segments was worse for afternoon procedures (P < 0.05 for some segments) and for patients with prior constipation (P < 0.05 for descending colon segments). Caecal intubation times were shorter for patients with diarrhoea and longer for female patients, who also had shorter withdrawal times. No correlation was found between the procedural end points (total duration, caecal intubation time and withdrawal times) and the timing of fasting. CONCLUSIONS: Quality of cleansing is significantly improved when bowel preparation is taken entirely the day prior to colonoscopy. Patients with prior constipation demonstrated poorer cleansing.

Characterization of active site structure in CYP121. A cytochrome P450 essential for viability of Mycobacterium tuberculosis H37Rv.

Mycobacterium tuberculosis (Mtb) cytochrome P450 gene CYP121 is shown to be essential for viability of the bacterium in vitro by gene knock-out with complementation. Production of CYP121 protein in Mtb cells is demonstrated. Minimum inhibitory concentration values for azole drugs against Mtb H37Rv were determined, the rank order of which correlated well with Kd values for their binding to CYP121. Solution-state spectroscopic, kinetic, and thermodynamic studies and crystal structure determination for a series of CYP121 active site mutants provide further insights into structure and biophysical features of the enzyme. Pro346 was shown to control heme cofactor conformation, whereas Arg386 is a critical determinant of heme potential, with an unprecedented 280-mV increase in heme iron redox potential in a R386L mutant. A homologous Mtb redox partner system was reconstituted and transported electrons faster to CYP121 R386L than to wild type CYP121. Heme potential was not perturbed in a F338H mutant, suggesting that a proposed P450 superfamily-wide role for the phylogenetically conserved phenylalanine in heme thermodynamic regulation is unlikely. Collectively, data point to an important cellular role for CYP121 and highlight its potential as a novel Mtb drug target.

Functional outcome and quality of life after ileal pouch-anal anastomosis in children and adults.

BACKGROUND: In the past, children with ulcerative colitis were treated with a total colectomy, ileostomy and mucous fistula; ileal pouch-anal anastomosis was postponed until adulthood. The aim of the present study was to assess the functional outcome and quality of life after ileal pouch-anal anastomosis and determine whether it is justified to perform the operation in children when surgery is indicated. METHODS: A retrospective review of 38 medical records was carried out, of which there were 19 paediatric patients and 19 adult patients (control). A questionnaire survey was conducted. Telephone interviews were carried out for the non-respondents. RESULTS: Sixteen patients in the paediatric group (nine boys, mean age: 12 years) and 16 patients in the adult group (10 men, mean age: 39 years) were available for analysis. There was no operative mortality. The mean bowel frequency per week was 37 and 42. Furthermore, bowel frequency during the day was slightly lower in the paediatric group. Children had marginally better continence than adults. In the quality of life assessment, the mean utilities in the paediatric group were 0.69 and 0.84 in the preoperative and postoperative status, respectively. These were similar to those in the adult group (0.62 and 0.82). Both groups achieved significantly favourable postoperative responses in terms of ability to perform social activity, recreation and enjoying food. CONCLUSIONS: Ileal pouch-anal anastomosis in children is safe, results in good functional outcome and improves the quality of life. Hence, it is justified to perform ileal pouch-anal anastomosis as soon as surgery is indicated rather than as a delayed procedure.

Cholecystectomy in patients with Crohn's ileitis.

PURPOSE: Gallstone disease is reported to be higher in patients with Crohn's disease than in the general population. This study was designed to determine the prevalence of cholecystectomy in patients with Crohn's ileitis, attempt to identify any associated risk factors, and determine whether it is justified to perform prophylactic cholecystectomy during ileocolic resection. METHODS: A total of 191 patients with Crohn's ileitis who were treated medically or who had an ileocolic resection were retrospective reviewed. A questionnaire survey was performed. Telephone interviews were conducted for the non respondents. Further review of medical records was performed to determine the details of admissions for any gallstone disease and/or subsequent cholecystectomy. A control group matched for age and gender was obtained. RESULTS: A total of 191 questionnaires were mailed, and the overall response rate was 70.2 percent (134/191) after telephone interview follow-up. There were 2 of 45 medical and 18 of 89 surgical patients with symptomatic cholelithiasis, i.e., 14.9 percent (20/134) of respondents. As a result, 2 patients (1.5 percent) required endoscopic sphincterotomy, 17 patients (12.7 percent) needed cholecystectomy, and 1 patient (0.7 percent) did not have any intervention. Only five patients had a cholecystectomy after their ileal resections. In the control group of 150 patients, 15 patients (14 females; mean age, 51.9 years; range, 34-78 years) had previous cholecystectomy. There was no significant difference with prevalence of cholecystectomy in Crohn's patients compared with controls (17/134 vs. 15/150; P = not significant). Furthermore, the number of ileal resections did not affect the cholecystectomy rate, but patients who had >30 cm of ileum resected were more likely to have cholecystectomy (P = 0.056). CONCLUSIONS: The prevalence of gallstone disease in Crohn's ileitis requiring cholecystectomy is similar to that of the general population with a female predominance. In addition, the number of patients requiring cholecystectomy after ileal resection was low. Thus, synchronous prophylactic cholecystectomy during ileocolic resection for Crohn's ileitis is not justified.

Short-term and long-term results of combined sclerotherapy and rubber band ligation of hemorrhoids and mucosal prolapse.

PURPOSE: Rubber band ligation is a common office procedure for symptomatic hemorrhoids. The aim of the study was to assess our short-term and long-term results of combined sclerotherapy and rubber band ligation in the management of hemorrhoids and incomplete mucosal prolapse. METHODS: Data on 6,739 patients who had previous combined sclerotherapy and rubber band ligation by the senior authors (GLN and PRD) were retrieved from the database dating between January 1976 and June 2000. These patients either had hemorrhoids or incomplete mucosal prolapse. Furthermore, questionnaires were sent to a random sample of 2,400 patients. Telephone interviews were performed for 600 of the nonrespondents. RESULTS: Of 6,739 patients (3,683 males; mean age, 46.7 years) in the database, 4,686 (70 percent) received the procedure once, and 2,053 (30 percent) received the procedure more than once. There were 5,689 patients (84 percent) who had their procedures performed consecutively within a planned period, and only 1,050 patients (16 percent) had repeat procedures after a period of more than 12 months from their last treatments. Thus, the recurrence rate was 16 percent. The overall complication rate was 3.1 percent, with minor bleeding being the major complaint. With regard to the questionnaire, 44 percent responded. The mean follow-up period was 6.5 (range, 1-11) years. There were patients who had residual symptoms of bleeding (19 percent), itch (21 percent), and lump (20 percent). However, 58 percent of patients who replied were asymptomatic. With satisfaction scores ranging from +3 to -3 (+3 indicating complete satisfaction and -3 indicating complete dissatisfaction), 90 percent scored >/=1, 9 percent scored 0 or less, and 1 percent did not specify a score. Hemorrhoidectomy was required in 7.7 percent of the responders. Of 600 phone interviews with the nonrespondents, 152 responded to the questionnaires. Although there was less satisfaction from the phone respondents, which may have accounted for the initial nonresponse, no statistical difference was detected in residual symptoms. CONCLUSIONS: Combined triple sclerotherapy and rubber band ligation is an effective treatment for early hemorrhoids and incomplete mucosal prolapse, with low rates of recurrence, complications, and hemorrhoidectomy, and it can be repeated easily.