Repair of a huge vesicocutaneous fistula with the rectus femoris musculocutaneous flap and VAC.

Adequate drainage is a prerequisite for closing a vesicocutaneous fistula, but the continued exposure to urine makes this impossible. Here, VAC was successfully used as a preoperative therapy before flap transplantation.

Reduction of tumor necrosis factor alpha-inducing capacity of recombinant Lactobacillus casei via expression of Salmonella OmpC.

The insertion of a heterologous gene into commensal bacteria is a common technique to develop a delivery agent for vaccination and therapies, but the pleiotropic effects of genetic modifications need to be investigated before its use in practical applications. Although supplemental properties provided by the expression of heterologous antigens have been reported, the negative or side effects on the immune-modulating properties caused by recombination are barely understood. In the present study, we fortuitously found that the secretion of tumor necrosis factor alpha (TNF-alpha) from murine macrophages was reduced by recombinant Lactobacillus casei expressing Salmonella OmpC compared to the stimulation of TNF-alpha secretion by nonexpressing L. casei. This reduction could not be attributed to OmpC as a purified protein. The main component of the OmpC-expressing strain included in the attenuation of TNF-alpha release seemed to be the cell wall, which exhibited higher sensitivity against N-acetylmuramidase than that of nonexpressing strains. These results suggest that the recombinant strain expressing a specific heterologous antigen might be digested rapidly in macrophages and lose immune-stimulating capability at an early time point.

Display of heterologous proteins on the surface of Lactococcus lactis using the H and W domain of PrtB from Lactobacillus delburueckii subsp. bulgaricus as an anchoring matrix.

AIMS: The aim of this study was to develop a cell-surface display system for foreign antigens on the surface of a Lactococcus lactis strain using an H and W domain of PrtB from Lactobacillus delburueckii subsp. bulgaricus as an anchoring matrix. METHODS AND RESULTS: To construct a cell-surface display pACL1 vector, a derivative of pSECE1 vector, we amplified the H and W domain of the cell-surface proteinase Prt B from Lact. bulgaricus using specific primers and then cloned it into a site downstream of the secretion signal sequence in the pSECE1 vector. The new system, designed for cell-surface display of recombinant proteins on L. lactis, was evaluated by the expression and display of the FliC protein of Salmonella enterica serovar Enteritidis as a reporter gene (pALC1:FliC). The expression of the FliC protein by the transformed cells was analysed by Western blot analysis, and the localization of FliC on the cell surface was confirmed by immunofluorescence microscopy and flow cytometry analysis. A specific band corresponding in size (approx. 110 kDa) to FliC plus the anchor residues was detected by anti-FliC antibody in the cell extract of L. lactis H61 harbouring pALC1:FliC, but not L. lactis H61 harbouring pALC1. In addition, flow cytometry and immunofluorescence microscopy revealed FliC-specific positive signals and a significant increase of fluorescence, respectively, in cells harbouring pALC1:FliC compared with that in control cells harbouring the parental pALC1 plasmid. These findings demonstrated that FliC was successfully displayed on the cell surface by the anchor domain of PrtB. CONCLUSIONS: A pALC1 vector using the H and W domain of PrtB from Lact. bulgaricus as an anchoring matrix can be used to successfully display the FliC protein on the surface of L. lactis. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel way of displaying heterologous proteins on the cell surface of L. lactis using the PrtB anchor domain should prove useful for the delivery of antigens and other proteins.

A new umbilicoplasty for children: creating a longitudinal deep umbilical depression.

When forming a deep umbilical depression for paediatric patients displaying a thin layer of abdominal fat, the V-Y advancement flap method is useful. However, although many plastic surgeons approve of a longitudinal umbilical depression as an ideal, conventional flap methods often result in only a longitudinal scar line with a depression at the end of the scar. The shifted umbilical depression facing upward or downward appears aesthetically unnatural. To resolve these problems, we have devised a new method of umbilicoplasty. Using obliquely opposing flaps, a longitudinal deep umbilical depression facing forward can be created at the correct umbilical position.

Crab-pincers style facial artery flap.

To reconstruct defects as large as 5 cm in diameter in the region extending from the columella and anterior nasal floor to the upper lip, we use a crab-pincers style facial artery (CPFA) flap technique combining nasolabial flaps and cheek advancement flaps. In the CPFA flap, the bifurcation of the facial artery allows the angle between the nasolabial flap and the cheek advancement flap to be freely altered in the manner of crab pincers. By combining the four leaves of bilateral CPFA flaps at the centre, appropriate reconstruction of the three-dimensional structures surrounding the columella can be achieved. In addition, this method requires only one operation to complete extensive reconstruction. The method does not result in adverse scarring or scar contracture. After the procedure,appropriate moustache growth gives a natural impression and conceals philtral distortion. We consider the CPFA flap to be very useful for reconstructing large defects in the central facial region.

Effects of dietary bile acids on formation of azoxymethane-induced aberrant crypt foci in F344 rats.

The present study has demonstrated the influence of bile acids (BAs) on the development and growth of azoxymethane (AOM)-induced aberrant crypt foci (ACF). Male F344 rats were treated with two doses of AOM (15 mg/kg) at 7 days apart and fed either basal MF or MF plus 0.4% of cholic (CA), deoxycholic (DCA), chenodeoxycholic (CDCA), lithocholic (LCA) and ursodeoxycholic (UDCA) acid mixed diets for 8 weeks after the first AOM dose. The mean number of ACF/colon of the rats fed CA, DCA, CDCA and LCA were higher than that of MF-fed group and the differences were statistically significant (P < 0.005). But the mean number of ACFs/colon was significantly (P < 0.005) lower in UDCA diet-fed rats compared to MF. UDCA-fed rats also showed a significant decrease in average crypt multiplicity (number of crypts/focus) of ACF compared to MF alone. The mean number of ACF with > or =5 crypts was about 2.5-3.7 times higher in case of CA, DCA, CDCA and LCA and about 8.2 times lower in UDCA compared to the control MF diet group. In a parallel study, feeding for 18 weeks of the same BAs mixed diets without AOM administration did not significantly induce ACF. Therefore, these data suggest that dietary BAs by themselves do not induce ACF in F344 rats but enhance or, in the case of UDCA, suppress the development and growth of AOM-induced ACF.

DNA adduct formation by hormonal steroids in vitro.

We examined the binding of various steroid hormones to DNA in vitro by means of 32P-postlabeling. Seventeen steroid hormones and cholesterol (CS) were incubated with human liver DNA at 37 degrees C for 1 h under aerobic conditions in the absence of catalysis. The reaction mixtures were analyzed by the nuclease P-1 version of 32P-postlabeling. The results showed that cortexolone (CX), prednisolone (PS), cortisone (CN), cortisol (CL), tetrahydrocortisol (TC), corticosterone (CC), 11-deoxycorticosterone (DC), dexamethasone (DX), dihydrocortisol (DL), and aldosterone (AL) covalently bound with DNA. However, progesterone (PG), 17 alpha-hydroxyprogesterone (HG), estrone (E1), estradiol (E2), estriol (E3), testosterone (TS), cortol (CR) and the original compound for biosynthesis, CS, did not form adducts. In absence of DNA, the steroids themselves did not give rise to any spot on TLC under the same conditions. The dose-responses of DNA binding by DC, DL, CC, CL and CN were linear. The relative adduct labeling of reactive steroids at a concentration of 2 mM were as follows: 68.8 (CX), 53.2 (PS), 39.6 (CN), 29.9 (CL), 20.9 (TC), 12.9 (CC), 12.3 (DC), 7.5 (DX), 4.7 (DL), 1.2 (AL) adducts per 10(8) nucleotides. Reactive and nonreactive steroids were distinguishable by the presence or absence of the carbonyl group (-CO-CH2OH) at carbon seventeen (C17) of the cholesterol skeleton. This implies that the electrophilic carbonyl or a neighboring group perhaps involved in the formation of covalent bond with DNA. To investigate the nature of target base(s) of these DNA reactive steroids, mononucleotides of all four bases of DNA were reacted with CN, CL, CC and cochromatographed with the obtained spots of DNA reactions. The results of which stated that these steroids and guanine reaction gave the same spots as observed in DNA reaction, indicating guanine is the main target of these DNA reactive steroids. Hep G2 human hepatocellular carcinoma cells were used as an alternative model. Although nine steroids (CL, DL, TC, PS, DX, PG, E2, TX, CR) did not react with intracellular DNA under our experimental conditions, our findings suggested that some hormonal steroids can form covalent DNA adducts in vivo.

[A case of middle cranial fossa meningioma extending into the infratemporal fossa: an approach to the pterygoid extension of the sphenoid sinus via the infratemporal fossa].

A case of middle cranial fossa meningioma extending into infratemporal fossa and the pterygoid process is presented. The patient had a sphenoid sinus extending inferolaterally into the pterygoid process, which is known as pterygoid extension of the sphenoid sinus. This type of variation of the sphenoid sinus allowed a safe and well-oriented approach to the pterygoid process via the infratemporal fossa. The tumor extending into the pterygoid process was removed successfully without damaging any surrounding structures, e.g. maxillary nerve or Vidian nerve. However, pterygoid extension of the sinus is seen in only 40% of cases. Therefore, close preoperative examination with bone window CT scan is mandatory before employing this approach.

Secondary contour reconstruction of maxillectomy defects with a bone graft vascularized by flowthrough from radial vascular system.

We describe post-total maxillectomy secondary facial contour reconstruction using an osteocutaneous scapular flap nourished by flow-through vascularization from the radial vascular system. Scar contracture caused by either total or partial maxillectomy for maxillary cancer was completely released with exposure of the edge of the zygomatic arch, orbital floor, and nasal bone. The scapular skin flap was placed into the mucosal defect, and the orbital floor and zygomatic prominence were reconstructed with the scapular bone. The flap nutrient vessels were anastomosed to radial vessels and cephalic vein grafts. Two representative cases are illustrated to demonstrate the application and advantage of this operative method.

Mucosa-preferential DNA adduct formation by 2-amino-3-methylimidazo-[4,5-f]quinoline in the rat colonic wall.

The mechanism of mucosa-specific formation of DNA adducts, which was found recently in human intestines, was studied in male F344 rats treated with 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). There are three conceivable pathways for p.o. administered IQ to reach the target colonic mucosal cells: pathway 1, through the digestive canal which exposes from the lumenal direction; pathway 2, following enterohepatic circulation re-expose from the lumenal direction; and pathway 3, exposure via blood circulation. To investigate these possible pathways, the following surgical procedures were performed: (a) portal catheterization for IQ administration to eliminate pathway 1 and (b) choledochal catheterization for bile drainage to eliminate pathway 2. When both procedures are combined, only pathway 3 is active. Four types of IQ-DNA adducts were commonly observed in the colons of all experimental groups, with no qualitative difference between the mucosal and muscular layers. When IQ-HCl was administered by p.o. gavage at a dose of 100 mumol/kg body weight, approximately 70% of the IQ-DNA adducts in the colonic mucosa (13.1 +/- 4.3 adducts/10(7) nucleotides) was induced through pathway 1. Pathway 3 induced the remaining 30% of mucosal adducts, producing equal adduct levels in both layers. Pathway 2 did not work for adduct formation. The DNA adduct formation was unaffected in the presence of intestinal flora, indicating that detoxified IQ does not reactivate by floral enzymes. In conclusion, mucosa-specific DNA adduct formation in the colon is caused most likely by the absorption of carcinogens through the lumen.

Mucosa-specific DNA adducts in human small intestine: a comparison with the colon.

The presence of several covalent DNA adducts in the human colon was demonstrated by 32P-postlabeling in a previous study. We demonstrated that DNA of all the colonic mucosa tested were selectively adducted by a single genotoxic agent and this modification was completely absent in the DNA of muscular layers. In this study, the DNA adducts of the small intestine are compared with those of the colon to understand the role of mucosa-specific DNA adduct (MSA) in intestinal carcinogenesis. The mucosal DNA of the small intestine from 19 adults undergoing surgery due to gastric carcinoma (seven cases), pancreatic carcinoma (four cases), colon carcinoma (four cases), small intestinal tumor (two cases), intestinal trauma (one case) and ectopic pancreas (one cases) were analyzed. The mucosa-adjacent muscular layer DNA of the corresponding samples was examined as a control. Several common DNA adducts were observed in both mucosal and muscular layers of all the adults. Besides these common background DNA adducts, two MSAs (Si1, Si2) were detected in most of the adults as in colon cases. Si1 was present in all adults examined (19/19 cases) at a level of 0.04-0.22 adducts/10(8) nucleotides (average 0.09) and Si2 was found in 13/19 patients at a level of 0.03-0.08 adducts/10(8) nucleotides (average 0.05). Si2 was the same adduct detected in the adult colonic mucosa. However, they were absent in the adjacent muscular layer as well as in the neonatal intestine tested as a control. The total level of the mucosa-specific DNA adducts of the small intestine was approximately 28-fold lower than that of the colon. Considering that the incidence of cancer in the small intestine is rather lower than that in the colon, these results may be relevant to the development of human intestinal cancer.

[A case of umbilical polyp with aberrant pancreas and small intestinal mucosa--analysis of cases of umbilical polyp reported in Japan].

We report a case of a 60 year-old man with an umbilical polyp composed of the aberrant pancreas and small intestinal mucosa. This is an extremely rare lesion which originates in remnants of omphalomesenteric duct and is usually diagnosed in the infant period. Microscopic examination disclosed the aberrant pancreas with exocrine glands and ducts, but no islets of Langerhans. The above findings suggested that the pancreas might have been HEINRICH II in type.

In vitro formation of DNA adducts with bile acids.

The in vitro experiment was carried out following 32P-postlabeling analysis to determine the DNA-reactive bile acids present in the human body. The unconjugated and conjugated forms of cholic (CA), chenodeoxycholic (CDCA), deoxycholic (DCA) and lithocholic acid (LCA) were added to calf thymus DNA followed by 1 h of incubation at 37 degrees C. After the incubation the mixture was analyzed by the nuclease P1 modification of 32P-postlabeling. Among the 12 bile acids tested, our results showed that unconjugated CDCA and LCA and the glycine and taurine conjugates of LCA (g-LCA, t-LCA) were able to bind covalently with naked DNA in vitro without intervention of any catalyst. It was also shown that bile acid alone did not give any spot on TLC. These binding reactions depended on the bile acid concentration in a linear manner. The data on the extent of binding at a concentration of 0.1 mg/ml showed values of 28.5 (t-LCA), 23.7 (g-LCA), 3.47 (LCA) and 1.32 (CDCA) adducts per 10(8) nucleotides. These reactive bile acids were also incubated with COLO 205 human colon carcinoma cells and Hep G2 human hepatocellular carcinoma cells for 24 h, but no specific DNA adduct was formed. Further, when LCA or CDCA was administered into male Fischer 344 rats by gavage at a dose of 10 mg/rat every 8 h for 3 days, no specific DNA adduct was detected in their liver or colon. Covalent DNA adducts are believed to cause alteration of the primary structure of genes, which is potentially linked to carcinogenesis. Though our preliminary data failed to detect any bile acid-related DNA adducts in the cultured cells or in rats, the results may provide a basis for assuming some of these bile acids to be potential initiators of colon cancer.

Presence of mucosa-specific DNA adduct in human colon: possible implication for colorectal cancer.

DNA of normal mucosa and the adjacent muscular layer from 18 adults suffering from colorectal neoplasms was examined by 32P-post-labeling analysis in order to estimate the exposure of the human colon and rectum to environmental carcinogens. Colorectal DNA samples obtained from six newborns were also examined as a normal control because they were presumed to have been minimally exposed to environmental carcinogens. One common mucosa-specific DNA adduct was found in the normal colorectal wall in all adults at the level of 0.10-34.13 adducts/10(8) nucleotides (mean +/- SD: 3.64 +/- 7.92 adducts/10(8) nucleotides), however, these were absent from the newborns' colons. Although several common spots were present in the mucosa, muscular layer and newborn tissues, there was no muscular layer-specific DNA adduct. The relationship between the levels of the mucosa-specific DNA adduct in the non-cancerous part and the histological degree of malignancy was not significant. The presence of this mucosa-specific DNA adduct in adult colon suggests that the human colon is commonly exposed mainly to one environmental carcinogen. This carcinogen is supposed to originate from foods, because the incidence of colorectal carcinoma is closely linked to dietary habits and the mucosa-specific DNA adduct was not present in newborns who had never ingested food. The incidence of adult colonic cancer originating from its mucosa is high, while cases of muscular origin or in newborn colon are rare. Therefore, the mucosa-specific DNA adduct is presumably responsible for the development of colonic cancer of epithelial origin.

Repair of "coup de sabre" with tissue expander.

Two cases of coup de sabre, a linear form of scleroderma, treated by means of tissue expansion, are discussed. Expanded skin flaps on the scalp and forehead provided sufficient skin and tissue for closure of the defect after resection of the defect, and good results were obtained.

Dermatofibrosarcoma protuberans with regional lymph node metastasis: a case report.

The case of a 21-year-old man with dermatofibrosarcoma protuberans of the scalp that spread to deep cervical lymph nodes is presented. Available literature is reviewed, and the infrequency of metastasis is discussed. Since cases of metastases almost always involve recurrent lesions, the importance of wide and deep resection in the initial operation cannot be overemphasized.