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Stimulation of precise excision and recombination by conjugal proficient F'plasmids.

Large F plasmids such as F'128 stimulate precise excision of the transposons Tn5 and Tn10 in E. coli K12. This stimulation occurs when the transposons are either on the F'128 plasmid or the bacterial chromosome. Stimulation of precise excision is dependent upon conjugal transfer proficient F'plasmids. Tra- mutations which are defective in conjugal transfer negate this F'128 plasmid stimulation effect. F'128 traS mutations, which are surface exclusion defective and thus permit matings between male cells, thereby increasing conjugal transfer, increase the F plasmid stimulation effect. When the F' plasmid is present in a cell with the small plasmid, pRS31, carrying the traS to traZ region of F, stimulation of precise excision is no longer observed. This complementation-like activity by pRS31 is abolished by a Tn5 insertion in the traS gene. Data are presented supporting the notion that F' plasmid stimulation of precise excision occurs in the recipient during conjugal transfer. F'128 traS also stimulates recA-dependent recombination between DNA sequences on the small, nontransferrable plasmid pRDK41, DNA sequences that are unrelated to those of the F plasmid. The F'plasmid stimulation of precise excision of Tn5 is not seen with F+ but only with certain F's with large insertions of chromosomal DNA.

Conjugation, Genetic↗

Evidence in a nematode for regulation of transposon excision by tissue-specific factors.

The transposable element Tc1 in Caenorhabditis elegans undergoes an excision reaction, which can be detected in a Southern hybridization as the appearance of empty chromosomal insertion sites. This excision reaction is under tissue-specific regulation in that it occurs at much higher frequency in somatic cells than in the germ line. We show here that this regulation is likely to be due to the action of tissue-specific factors that either promote excision in somatic tissues or repress it in the germ line. The rate of excision of elements at five distinct chromosomal sites has been measured by a method that avoids ambiguities due to cell division. All these elements are found to undergo excision at closely similar rates during the L1 larval stage. No distinct difference exists among the elements at different sites that would suggest regulation by flanking sequences.

Animals↗

Identification and mapping of regions that confer plasmid functions and of sites for excisive recombination of plasmid pMMC7105.

Strain PP808 of Pseudomonas syringae pv. phaseolicola contains pEXC8080 (34.6 kb), the smallest of several plasmids that originated by partial excision of the cryptic plasmid, pMMC7105 (150 kb), from the host chromosome. This excision plasmid is derived entirely of sequences from pMMC7105 and contains a 24 kb region referred to as common DNA, which is present in each of the other excision plasmids. A six enzyme restriction endonuclease map was constructed of pEXC8080. The replication region was mapped by identifying small restriction fragments that conferred replication properties to pMB1 plasmids that otherwise fail to replicate in Pseudomonas. This region is located within the common DNA and is 0.8-3.8 kb in size. Sequences from pEXC8080 failed to stabilize pMB1 derivatives in Pseudomonas in the absence of antibiotic selection, but stability functions were mapped to a region of pMMC7105 that presumably remains integrated in the chromosome of strain PP808. An incompatibility region was mapped to a 7.3 kb region on pEXC8080 that is closely linked to, but not included within, the replication region. The recombination site was mapped to a 1.2 kb region of the fusion fragment that was formed upon excision of pEXC8080. RS-I, a repetitive sequence found on pMMC7105 was present in the fusion fragment at the site of recombination. RS-I was also mapped to BamHI fragments that recombined upon excision of pEXC8080 and suggest that it provides sites for homologous recombination.

Anti-Bacterial Agents↗

Enhanced frequency of transposition of the maize transposable element Activator following excision from T-DNA in Petunia hybrida.

Many of the systems currently employed for heterologous transposon tagging in plants rely on an excision assay to monitor transposon activity. We have used the streptomycin phosphotransferase (SPT) reporter system to assay Ac activity in Petunia hybrida. In other species, such as tobacco or Arabidopsis, excision of Ac from the SPT gene in sporogenous tissue gives rise to streptomycin-resistant seedlings in the following generation. The frequency of fully streptomycin-resistant seedlings in petunia was low (0.4%) but molecular analysis of these indicated that the actual excision frequency may be as low as 0.05%. This indicates that the SPT assay is not a reliable selection criterion for germinal excision in petunia. Extensive molecular screening for reinsertion of Ac was consistent with a low primary transposition frequency (0%-0.6%). In contrast to these findings, the progeny of confirmed germinal transpositions for three independent transformants showed frequent transposition to new sites (9.5%-17.0%). This suggests a high frequency of secondary transposition compared with primary transposition from the T-DNA. Segregation analysis indicates that the high transposition activity is closely associated with transposed copies of Ac. No evidence was found for an altered methylation state for Ac following transposition. The implications of these results for heterologous transposon tagging in petunia are discussed in the context of the reliability of excision reporter systems in general.

DNA Repair↗

Reconstruction with bilateral gluteus maximus myocutaneous rotation flap after wide local excision for perianal extramammary Paget's disease. Report of two cases.

PURPOSE: Extramammary Paget's disease is a rare dermatosis. Wide local excision is recommended in patients with perianal extramammary Paget's disease. After wide local excision, it is necessary to do reconstruction, with preservation of bowel function. We present here two cases of perianal Paget's disease, in which the patients were treated by reconstruction with bilateral gluteus maximus myocutaneous rotation flap after wide local excision. PATIENTS AND METHODS: A 55-year-old woman and 58-year-old man were admitted with anal pain and bleeding. Histologic examination of the perianal lesion revealed the presence of typical Paget's cells, and no underlying carcinoma and no distant metastasis was detected in either patient. Wide local excision, including the rectal mucosa, was performed, with reference to intraoperative frozen sections. Surgical defect was overlapped with bilateral gluteus maximus myocutaneous rotation flap, and the anus was reconstructed. RESULTS: Postoperative bowel function and quality of life were well preserved, and flaps healed satisfactorily. Patients have had no recurrence and have been able to return to work. CONCLUSION: Bilateral gluteus maximus rotation flap may be useful after wide local excision of perianal extramammary Paget's disease without underlying invasive carcinoma.

Anal Canal↗

Local recurrence after curative resection of cancer of the rectum without total mesorectal excision.

PURPOSE: The aim of this article was to examine local recurrence after curative resection for carcinoma of the rectum in which the surgical technique of total mesorectal excision was not performed. METHODS: A single surgeon managed the patients and the data collected prospectively. Total excision of the distal mesorectum was not performed in the upper third or mid rectum. RESULTS: From 1969 to 1993 curative resections were performed in 549 patients, of which 17 died postoperatively, leaving 532 for analysis. Sphincter-saving resection was performed in 468 patients (88 percent) and abdominoperineal excision in 58 (10.9 percent). The pathology stages (Dukes) were A, 158 (29.7 percent); B, 184 (34.7 percent); and C, 190 (35.7 percent). Five hundred seventeen patients (97.2 percent) were followed up for a minimum of five years. The median period of follow-up was 82 months. Local recurrence confined to the pelvis occurred in 17 patients, and local recurrence associated with distant metastases occurred in 24 patients. The total five-year local recurrence rate was 7.6 percent. Local recurrence was increased in Stage C tumors (P = <0.0001). Diathermy dissection in the pelvis was associated with a decreased local recurrence rate (P = 0.023). The five-year survival rate in curative resections was 72.5 percent. CONCLUSIONS: It is essential that articles presenting local recurrence rates should include both local recurrence in isolation and that which occurs with distant metastases. Although total mesorectal excision for rectal cancer was not performed in this study, the local recurrence rate is not materially different from that in several articles where total mesorectal excision has been used. Whether the distal mesorectum needs to be pursued in mid-rectal cancer is not yet proven.

Adult↗

Staged excision and split-thickness skin graft for circumferential perianal Paget's disease.

PURPOSE: This study was designed to describe a surgical technique for the treatment of circumferential Paget's disease. METHODS: A search of our computerized patient registry was undertaken, and case records for those patients with perianal Paget's disease were systematically reviewed. RESULTS: Between July 1993 and October 1998, four patients with perianal Paget's disease were identified. Circumferential lesions were identified in three of the four patients. Wide local excision was considered the procedure of choice. All patients underwent a two-staged excision using split-thickness skin graft reconstruction. A similar second stage was performed six to eight weeks later; the other half of the circumference was excised and grafted. No patient had a protective stoma. Graft survival was 100 percent for two patients (four operations) and 80 percent and 70 percent for the other two patients (two operations). The remaining surface healed successfully by secondary intention. In one patient, residual disease was positive at one margin, and a third local excision and split-thickness skin graft was performed. Hospital stay ranged from five to nine days for each procedure. There were no major complications; one patient developed a mild anal stenosis three months after the second procedure and was successfully medically treated. CONCLUSIONS: Staged excision and split-thickness skin graft is a viable option for the treatment of circumferential perianal lesions. It carries a minimal morbidity and no observed mortality, the functional result is good, and it is technically simple compared with myocutaneous grafts. Moreover, a stoma is not required.

Aged↗

Excision of trocar sites reduces tumor implantation in an animal model.

PURPOSE: The purpose of this study was to determine the effect of excising abdominal trocar wound sites after pneumoperitoneum on the rate of trocar site tumor implantation in a hamster model. This would help determine whether tumor cells seed trocar sites during or after pneumoperitoneum. METHODS: A total of 0.5 ml of GW-39 human colon cancer cell suspension at 2.5 percent v/v (8 x 10(5) cells) was injected into the abdomens of 77 hamsters through a midline incision. Animals were subjected to ten minutes of pneumoperitoneum, after placement of four abdominal trocars, and then randomly assigned to undergo either simple suture closure or 4-mm radius trocar wound site excision at the end of the procedure. Gross and microscopic tumor implants were documented seven weeks later. RESULTS: There were three and four deaths in simple suture closure and wound site excision groups, respectively. Of the remaining 35 hamsters in each group, tumor cells implanted at 89 and 78 percent of trocar sites, respectively (P < 0.03). There was no significant difference between the two groups in tumor implantation at midline laparotomy sites. Wound site excision also resulted in fewer palpable tumors (44 vs. 61 percent; P < 0.01) and a lower tumor implantation rate (49 vs. 74 percent; P < 0.05) at all four concurrent sites compared with simple suture closure. CONCLUSIONS: Excision of laparoscopic abdominal trocar wound sites can significantly, but not completely, reduce tumor implantation rate compared with simple wound closure.

Abdominal Muscles↗

[Selection of patients with rectal tumors for local excision based on preoperative diagnosis. Results of a consecutive evaluation study of 552 patients].

INTRODUCTION: The aim of our study was to evaluate the accuracy of clinical staging (CS), biopsies, and endoluminal ultrasonic examination (EUS) in preoperative staging of rectal tumors treated with transanal local excision. This local excision is an adequate procedure for benign rectal polyps and low-risk T1 carcinoma. PATIENTS AND METHODS: The study included 552 patients with rectal adenocarcinoma, villous adenoma, or tumors with other histologic characteristics who underwent a transanal excision (transanal endoscopic microsurgery n=513 or transanal excision n=39). We compared the results of CS, biopsies, and EUS with postoperative pathology findings. RESULTS: Preoperative histological diagnosis of the rectal carcinoma depended on tumor size (52% in cancers <3 cm, 25% in cancers >3 cm; p=0.001) and was correct in 56% of cases. Transanal ultrasonography (uT0/1) had superior sensitivity (95% vs 78%) and a higher positive predictive value (93% vs 85%) than clinical staging (CS I) in detecting adenoma or T1 rectal carcinoma, whereas specificity was similar in both (62% vs 58%). In patients in whom preoperative histological analysis revealed adenomas, transanal ultrasonography was accurate (uT0/1) for the postoperatively assessed adenoma pT1 in 97%, whereas diagnosis (uT0/1) was correct in only 71% of cases in which preoperative histological analyses showed rectal carcinomas. CONCLUSIONS: In patients with rectal tumors, preoperative staging with transanal ultrasonography and biopsy is essential for the indication and allows selection of patients for transanal local excision.

Adenocarcinoma↗

Cre/lox-mediated marker gene excision in transgenic maize (Zea mays L.) plants.

After the initial transformation and tissue culture process is complete, selectable marker genes, which are used in virtually all transformation approaches, are not required for the expression of the gene of interest in the transgenic plants. There are several advantages to removing the selectable marker gene after it is no longer needed, such as enabling the reuse of selectable markers and simplifying transgene arrays. We have tested the Cre/ lox system from bacteriophage P1 for its ability to precisely excise stably integrated marker genes from chromosomes in transgenic maize plants. Two strategies, crossing and autoexcision, have been tested and demonstrated. In the crossing strategy, plants expressing the Cre recombinase are crossed with plants bearing a transgene construct in which the selectable marker gene is flanked by directly repeated lox sites. Unlike previous reports in which incomplete somatic and germline excision were common, in our experiments complete somatic and germline marker gene excision occurred in the F(1) plants from most crosses with multiple independent Cre and lox lines. In the autoexcision strategy, the cre gene, under the control of a heat shock-inducible promoter, is excised along with the nptII marker gene. Our results show that a transient heat shock treatment of primary transgenic callus is sufficient for inducing cre and excising the cre and nptII genes. Genetic segregation and molecular analysis confirmed that marker gene removal is precise, complete and stable. The autoexcision strategy provides a way of removing the selectable marker gene from callus or other tissues such as embryos and kernels.

Chromosome Segregation↗

Total mesorectal excision for rectal cancer: the truth lies underneath.

The surgical technique itself has emerged as a crucial factor for local recurrence since the popularization of total mesorectal excision for the treatment of rectal cancer. This procedure is associated with lower local recurrence rates after "curative" surgery compared to traditional dissection of the rectum. The aim is to remove an intact mesorectal envelope from the promontorium down to the pelvic floor by sharp dissection with tumor-free margins and without causing injury to the pelvic nerves. However, the description of total mesorectal excision has been confusing. Moreover, the implication that total excision of all the perirectal fat contained within the perirectal fascia en bloc in all patients with rectal cancer can minimize local recurrence remains contentious. Therefore a critical appraisal of the procedure is required. Nonrandomized clinical studies have shown that total mesorectal excision reduces the local recurrence rate and increases disease-free survival in patients with adenocarcinoma of the middle and distal third of the rectum. Circumferential resection margins of 2 mm or more are associated with a lower local recurrence rate. Additional benefits in local control can be obtained with neoadjuvant treatment. Thus the modern treatment of rectal cancer combining total mesorectal excision with neoadjuvant chemoradiation results in excellent local tumor control. However, it is achieved at the cost of significant functional sequelae and impaired quality of life. The development of therapeutic alternatives that can achieve similar rates of local and distant tumor control without the mortality, morbidity, and functional consequences of radical surgery is a major challenge for colorectal surgeons.

Adenocarcinoma↗

Local excision of rectal cancer.

The prospect of treating a rectal cancer often leads to significant fear among patients because of the possibility of a permanent colostomy. Although radical resection, in particular abdominoperineal resection, has been used effectively for rectal cancer treatment, other techniques such as local excision with or without adjuvant therapy have been used with significantly less morbidity than that of the abdominoperineal resection, with excellent cure rates. There are essentially three local excision techniques that can be used to remove a small rectal cancer completely. Selection criteria are critical for choosing the appropriate patient for these techniques. Tumors must be less than 4 cm in diameter and take up less than 40% of the rectal wall circumference. They also need to be relatively close to the dentate line and have no evidence of any invasion into the mesorectum or perirectal nodes. Preoperative staging with endorectal ultrasonography, computed tomography, and digital examination helps select appropriate patients. Retrospective series have shown significant success using local excision techniques, with local recurrence rates ranging from 0% to 11% for early-stage lesions. Prospective series have shown similar recurrence rates. Postoperative function with or without adjuvant therapy has not been adequately documented along with quality of life and must be part of any future reports on local excision techniques as well as all rectal cancer treatment studies. Local excision does, however, seem to provide adequate treatment in well selected patients and provides a less morbid alternative to the treatment of rectal cancer than radical resection, particularly abdominoperineal resection, which obligates the patient to a permanent colostomy.

Clinical Trials as Topic↗

Does follicle excision always result in enlargement of offspring size in lizards?

An experimental reduction of offspring number has been reported to result in enlargement of offspring size in lizards. We applied the "follicle excision" technique to a lacertid lizard (Takydromus septentrionalis) to examine whether this effect is generalisable to lizards. Of the 82 females that produced 3 successive clutches in the laboratory, 23 females underwent follicle excision after they oviposited the first clutch. Follicle excision reduced clutch size, but did not alter egg size. This result indicates that egg size is not altered during vitellogenesis in T. septentrionalis. Females undergoing follicle excision produced a third clutch (a second post-surgical clutch) as normally as did control females. Females switched from producing more but smaller eggs early in the breeding season to fewer but larger eggs later in the season. Our results indicate that female T. septentrionalis maximize reproductive success by diverting an optimal, rather than a higher, fraction of the available energy to individual offspring. This optimized allocation of the available energy to offspring production explains why follicle excision does not result in enlargement of egg size in this species. Our study provides evidence that an experimental reduction of offspring number does not always result in enlargement of offspring size in lizards.

Animals↗

Complications and recurrence after excision of rectal internal mucosal prolapse for obstructed defaecation.

BACKGROUND: Rectal internal mucosal prolapse (RIMP) may cause obstructed defaecation and encouraging short-term results have been reported after its transanal excision. The objective of this retrospective study was to assess both clinical and functional outcome after this procedure alone for patients presenting with evacuatory difficulty. PATIENTS AND METHODS: Forty patients (30 females, mean age 54 years), all suffering from obstructed defaecation, underwent RIMP excision at our unit during the last 11 years. RIMP was of first degree in three patients, of second degree in 21, and of third degree in 16 with 28/40 cases (70%) having associated anorectal pathology. The operation was carried out by hand suture (submucosal excision, Sarles endorectal excision, or the Delorme mucosectomy) in 26 patients, by circular stapled prolapsectomy in nine patients, or by combined manual and stapled techniques in five cases. Proctoscopy was carried out after 2 months for all patients, with anorectal manometry in 30 patients. Patients were independently assessed by state-trait anxiety scales for attendant anxiety and depression. RESULTS: Eighteen patients (45%) had significant postoperative complications with a surgical reintervention rate of 32.5%. Overall, 21 patients (52%) reported recurrent constipation and of these 14 (65%) had recurrent RIMP; six patients were treated successfully by rubber-band ligation alone. Two patients (5%) experienced new onset faecal incontinence. The recurrence rate of RIMP was unaffected by the type of operation, being 53% after manual techniques and 48% after combined procedures. There was no difference between postoperative manometric values in patients presenting with recurrent RIMP or constipation compared with those without RIMP or constipation on follow-up. Forty-eight percent of the patients with both recurrent constipation plus RIMP had manometric evidence of non-relaxing puborectalis syndrome compared with 26% with RIMP but without constipation (P<0.05). Ten of the 14 patients (71%) with anxiety and/or depression complained of recurrent constipation after surgery compared with nine of the 26 patients (24%) with normal psychological profiles (P<0.01). Patients with a preoperative rectocele were more likely to suffer from recurrent constipation than those without rectocele (eight out of 15, 53.3% vs. seven out of 25, 28%; P<0.05). CONCLUSIONS: Primary excision of RIMP does not seem an effective treatment for obstructed defecation with predictive factors for an adverse outcome in terms of recurrence (RIMP and constipation) including the presence of preoperative non-relaxing puborectalis syndrome and a demonstrated anxiety or depression psychological profile. The technique of prolapsectomy does not seem to affect outcome.

Adult↗

Clinical significance of a wide excision policy for sacrococcygeal chordoma.

PURPOSE: Surgical treatment of sacrococcygeal chordoma is very difficult and the recurrence rate is high. We investigated the outcomes of wide excision in 12 sacrococcygeal chordoma patients treated between 1983 and 2003. METHODS: The 12 patients underwent primary wide excision. The conventional procedure was wide excision employing a chisel and airtome for sacral tumours, and in 1997 a new threadwire saw procedure was introduced. All 12 patients were included in a retrospective analysis of tumour- and treatment-related variables, and outcomes. RESULTS: Six patients underwent surgery with a combined anterior-posterior approach at the cephalad sacral border. In 12 patients, 11 lesions were broadly excised, and in the 1 remaining patient the intralesional margin showed tumour contamination due to a fracture macroscopically. The latter patient received post-operative radiation therapy, but suffered a recurrence 8.0 years later. Three patients died before final follow-up: one from subsequent metastases, two elderly patients from complications. For the 10 chordoma patients, i.e. excluding the two who died due to complications, the 5, 10 and 20 years recurrence rates were 100, 66.7 and 66.7%, respectively. For the 12 patients, overall 5, 10 and 20 years survival rates were 83.3, 55.6 and 55.6%, respectively. CONCLUSIONS: Our results suggest that large chordoma should be widely excised, using a modified threadwire saw, with a combination of anterior-posterior procedures.

Adult↗

Analysis of the cis-acting DNA elements required for piggyBac transposable element excision.

The terminal DNA sequence requirements for piggyBac transposable element excision were explored using a plasmid-based assay in transfected, cultured insect cells. A donor plasmid containing duplicate 3' piggyBac terminal inverted repeats was constructed that allowed individual nucleotides or groups of nucleotides within one of the 3' repeats to be mutated. The relative extent of excision using the mutated end versus the wild-type end was then assayed. Removal of even one of the terminal 3' G nucleotides from the piggyBac inverted repeat, or removal of the dinucleotide AA from the flanking TTAA target site prevents excision of piggyBac at the mutated terminus. Incorporation of an asymmetric TTAC target site at the 3' end does not prevent excision from the mutated end. Thus, both piggyBac DNA and flanking host DNA appear to play crucial roles in the excision process.

Animals↗

Sac excision is essential to adequate laparoscopic repair of paraesophageal hernia.

BACKGROUND: We compared the incidence of early hernia recurrence in nonrandomized but consecutive patients undergoing laparoscopic repair of paraesophageal hernia (LRPH) without and with excision of the hernia sac. METHODS: LRPH was completed in 55 of 58 patients. In the first 25 patients, the sac was not excised. Total sac excision was performed in the subsequent 30 patients. All patients had crural repair with or without fundoplication, or gastropexy. RESULTS: Mean age of patients was 68 years (range, 34-95). There were three conversions; one patient died postoperatively. Mean operative time was 225 min in the first group and 190 min in the sac excision group. Median length of stay was 2 days (range, 1-15) for both groups. CONCLUSIONS: A precise method of total sac excision simplified dissection. It also ensured complete reduction of the hernia and availability of adequate esophageal length. Operative time was not increased, and no subsequent early recurrences were observed (p < 0.05).

Adult↗

Results of complete hemivertebra excision followed by circumferential fusion and anterior or posterior instrumentation in patients with type-IA formation defect.

To evaluate the results of surgical treatment in patients with unlocked full-segmented hemivertebra treated by excision. Twenty-six patients with a mean age of 12.4+/-1.7 years were included in the study. The mean duration of follow-up was 47.8+/-21.9 months. Diagnosis of type-IA hemivertebra was established by clinical, radiological, CT, and MRI evaluation. Preoperatively, patients were randomly allocated into two groups. In the first group, patients underwent anterior hemivertebrectomy initially; this was followed by posterior excision of the hemivertebra, posterior instrumentation, and fusion. In the second group, posterior components of the hemivertebra were excised at first, then the hemivertebra body was excised anteriorly, and this was followed by anterior instrumentation and fusion. For both groups, compression was applied to the convex side while distraction was applied to the concave side. Frontal and sagittal plane analysis of radiograms obtained preoperatively, postoperatively, and after a minimum period of 2 years was performed. The balance was analyzed clinically and radiologically by the measurement of the lateral trunk shift (LT) and shift of head (SH). The mean preoperative and postoperative Cobb angles were 45.5 degrees -/+11.4 degrees and 16.8 degrees -/+7.9 degrees, respectively, and postoperatively, a mean correction rate of 64.4+/-13.9% was obtained (P=0.00). The mean correction rate was 61.2+/-13.3% (19.2 degrees -/+7.6 degrees) for the last follow-up visit. Sagittal plane analysis demonstrated either conservation of physiological sagittal contours or a normalizing effect following excision of hemivertebra combined with anterior or posterior instrumentation. When postoperative balance values were compared, a statistically significant correction was found in terms of LT and SH values. Although none of the patients had complete balance (SH: 0 mm) or balanced curves (0 mm<SH<15 mm) preoperatively, 20 (76.9%) of the patients had a balanced trunk after surgical intervention. Circumferential fusion could be achieved in all cases. No neurological complication developed, the only complication was delayed wound healing. In view of these data, it is concluded that these techniques can be safely used for this patient group at low thoracic, thoracolumbar, and lumbar levels of vertebral column with high correction rates.

Adolescent↗