Induction of the suicide HSV-TK gene by activation of the Egr-1 promoter with radioisotopes.

In investigating new methods for the treatment of pancreatic cancer, we have explored the possibility of using a combination of radiation and gene therapies. We demonstrate herein that the early growth response gene 1 promoter (Egr-1) is sufficient to confer selective expression of the luciferase gene (Luc) in a human pancreatic tumor cell line (AsPc-1) when exposed to ionizing radiation. The Egr-1 promoter directed the radioinducible expression of luciferase, and yielded higher levels of Luc activity than that in nonirradiated lines. The radioisotopes Tc-99m, I-131, and Ga-67-citrate were selected as Egr-1 activators for their potential to accumulate in tumors. We studied Ga-67-citrate, a radioisotope employed in tumor scintigraphy, for its suitability for selective gene induction. The plasmid vector pEgr-1-Luc was transfected into AsPc-1 cells and then exposed to radioisotopes. Luciferase activity increased by 100-300 times over control. We also inserted the herpes thymidine kinase gene (TK) downstream of Egr-1 and transfected this construct into AsPc-1 cells. Ga-67-citrate and ganciclovir were added to the cells and cell survival was assessed by MTT assay. The growth of AsPc-1 cells transfected with the pEgr-TK construct was suppressed 2 days after exposure of the cells to Ga-67-citrate. The results indicate that Ga-67-citrate may be useful in combining radiation and gene therapies.

De novo glucan synthesis by mutants streptococcal glucosyltransferases present in pellicle promotes firm binding of Streptococcus gordonii to tooth surfaces.

Adherence of 3H-labelled cells of Streptococcus gordonii and Streptococcus milleri to artificial pellicles prepared from saliva supplemented with glucosyltransferases from mutants streptococci was examined using a new assay for sucrose-dependent cell-to-pellicle attachment. Results indicate that S. gordonii, but not S. milleri, could attach tightly to hydroxylapatite surfaces through de novo glucan synthesis by mutants streptococcal glucosyltransferases present in the experimental salivary pellicles.

Evaluation of three individual glucosyltransferases produced by Streptococcus mutans using monoclonal antibodies.

We previously established murine hybridomas producing a monoclonal antibody monospecific against three glucosyltransferases (I, SI and S) of Streptococcus mutans which contribute to dental caries formation. Here, we developed a new immunochemical technique (cross-dot system) with which individual levels of glucosyltransferases expressed by S. mutans can be evaluated. We also examined glucosyltransferase production and in vitro artificial plaque formation by a reference strain and several clinical isolates of S. mutans. The findings indicate that the levels of glucosyltransferases produced greatly vary with the cells and the culture medium, and that the cells producing high levels of both glucosyltransferase-SI and glucosyltransferase-I enzymes may possess high in vitro artificial plaque forming ability. We suggest that the cross-dot system will be useful for estimating the cariogenic potential of S. mutans isolates.

Hypertrophy of labia minora in myelodysplastic women. Labioplasty to ease clean intermittent catheterization.

Female myelodysplastic patients are sometimes annoyed by hypertrophy of the labia minora, possibly caused by long-term diaper dermatitis. This condition makes intermittent catheterization difficult and impairs the cleanliness and aesthetic appearance of the external genitalia. We successfully performed labioplasty on 3 myelodysplastic females.

Degradation kinetics and isomerization of cefdinir, a new oral cephalosporin, in aqueous solution. 1.

Hydrolytic degradation products of cefdinir were studied in acidic (pH 1), neutral (pH 6), and basic (pH 9) solutions. Seven major degradation products were isolated by preparative and/or high-performance liquid chromatography and characterized by UV, IR, 1H-NMR, and mass spectra. To clarify degradation pathways in each pH solution, kinetic and product analyses during hydrolysis of cefdinir were carried out along with the followup reaction of representative degradation products. Cefdinir was shown to degrade via two major degradation routes: beta-lactam ring-opening and pH-dependent isomerizations (lactonization, epimerization at C-6 or C-7, syn-anti isomerization of N-oxime function).

Degradation kinetics and isomerization of cefdinir, a new oral cephalosporin, in aqueous solution. 2. Hydrolytic degradation pathway and mechanism for beta-lactam ring opened lactones.

Hydrolysis of cefdinir leads to pH-dependent isomerizations and beta-lactam ring-opening. Lactam ring opened gamma-lactones were produced as a mixture of four diastereoisomers based on the lactone methyl, and C-6 isomerizations in acidic to neutral solutions. Cefdinir and its 7-epimer were hydrolyzed to clarify the pathway leading to these lactones and the mechanism of C-6 epimerization with the aid of chiral separation techniques. Chiral separation using a bovine serum albumin column was employed to detect the beta-lactam ring opened products of cefdinir and its 7-epimer; the C-6 and C-7 isomerization was thereby observed; however, it was found to be pH-dependent at pH > or = 9. Optical activity detection applied to the lactones produced from cefdinir and its 7-epimer demonstrated that the corresponding peaks of these lactones were enantiomeric pairs. In addition, the smallest rate constant at pH 4 was observed for C-6 epimerization of the lactones, and it was found to proceed without deprotonation at C-6 by 1H-NMR spectroscopy. From the results of these studies, a plausible mechanism for C-6 epimerization has been proposed. Additionally, it was confirmed that two degradation pathways were involved during hydrolysis of cefdinir to the lactone.

Degradation kinetics and mechanisms of a new cephalosporin, cefixime, in aqueous solution.

Hydrolysis of cefixime in buffer solutions (pH 1-9) at 25 degrees C and a constant ionic strength of 0.3 was investigated using ion-pair reversed-phase HPLC. Hydrolysis rates followed pseudo first-order kinetics; the rate of hydrolysis of cefixime was very slow at pH 4-7, slightly faster at lower pH, and quite rapid at higher pH. In the early stages of hydrolysis, six major degradation products were isolated and identified: a beta-lactam ring-opened product and a 7-epimer (basic conditions), three lactones derived from intramolecular cyclization between the 2-carboxyl and 3-vinyl groups (acidic conditions), and an aldehyde derivative involving a 7-acyl moiety (neutral conditions). Principal degradation pathways for cefixime were found to involve initial cleavage of the beta-lactam ring.

Tautomeric phenomenon of a novel potent immunosuppressant (FK506) in solution. I. Isolation and structure determination of tautomeric compounds.

Tautomeric phenomenon was observed in the HPLC chromatogram obtained from a novel potent immunosuppressant, FK506, in ethanol solution. Two tautomeric compounds derived from FK506 were isolated and purified by HPLC. Their structures were elucidated by spectral analyses. The mechanisms for this tautomerization were also established based on the results of structure analysis.

Effect of water content in perchloric acid on the non-aqueous potentiometric titration of nitrogen-containing compounds.

In the United States Pharmacopeia (USP), 0.1 N perchloric acid in acetic acid volumetric solution (hereafter HClO4 VS) used for non-aqueous titration has specified a water content between 0.02 and 0.05%. Preparing this titrant with such a narrow range of water content is very time consuming, precludes the use of commercially available titrants, and, consequently, prompted an investigation to try and expand the range up to 0.5%. In this study, the titrimetric results obtained using HClO4 VS containing more water were very close to those obtained using the USP specified titrants. A maximum assay difference of 0.7% in the titrations of three selected nitrogen-containing compounds, clonidine hydrochloride, dipyridamole, and adenosine were observed. The titrimetric results obtained using these titrants were also precise with RSDs of not more than 0.4%. Therefore, a wider range of water content in HClO4 VS between 0.02 and 0.5% is suggested for the USP potentiometric titration of nitrogen-containing compounds.

Method development for the determination of cefdinir and its related substances by high-performance liquid chromatography.

The validation of the HPLC method used for the determination of cefdinir and its related substances is described. The developed method was specific and stability-indicating and provided a linear response with concentration. The system and method precision, expressed as relative standard deviations, were not greater than 1%, and the reproducibilities within and between laboratories were acceptable for the assay method. The procedure can quantitate related substances greater than approximately 0.05% of the principal cefdinir peak.

Reverse-flow island flap: clinical report and venous drainage.

Twenty-two reverse-flow island flaps were transferred. These included peroneal, forearm, anterior tibial, and temporal flaps. Sixteen of 22 flaps survived completely. We encountered partial necrosis in 4 flaps and total necrosis in only 2 flaps. We credit this success to the reliability and availability of the peroneal, forearm, and temporal flaps; but we do not encourage use of the anterior tibial flap. The flaps that survived well did not show any signs of venous congestion. The advantage of the reverse-flow island flap is that it can be transferred from a proximal to a distal location. Using cadavers and fresh amputated limbs, studies on venous drainage of the reverse-flow island flap were performed. The venae comitantes had numerous venous valves and communicating branches, but more than sufficient reflux of the venous blood occurred through the valves at pressures of 90 to 105 cmH2O. We believe that the venous drainage of the reverse-flow island flap occurs as a result of reflux actions at the valve, communicating branches between the venae comitantes, and bypass vessels around the valves.

[Two cases of empyema treated with rectus abdominis myocutaneous flap and muscle flap].

A rectus abdominis myocutaneous flap (RAM flap) has often been useful in breast reconstructions after the surgical therapy for breast cancer, or in other plastic surgeries. However, there have been few reports concerning about its applications for intrathoracic diseases. In this paper, we report two cases of recurrent empyema treated with RAM flap and muscle flap from rectus abdominis. One case is 75-year-old male with right-empyema and bronchial fistula after the thoracoplasty due to lung tuberculosis. The bronchial fistula was successfully closed with muscle flap from the right rectus abdominis. The other is 33-year-old male with left empyema and cutaneous fistula after the hemothorax due to traffic accident. He contracted with MRSA in his thoracic cavity. We resected the fistula and reconstructed his left chest wall with left RAM flap. The muscle flap from rectus abdominis or RAM flap has been very effective for the therapy of bronchial fistula after lung tuberculosis, and for the reconstruction of chest wall.

[Chest wall reconstruction after the resection of primary chest wall tumors].

Between April 1983 and June 1994, we performed chest wall reconstructions on 9 patients (7 males and 2 females) out of 27 all of whom underwent the radical resection for the primary chest wall tumors. Histological diagnosis of tumors included 6 chondrosarcomas, 1 Ewing's sarcoma, 1 plasmacytoma and 1 giant cell tumor. Six lesions were located on the rib and 3 on the sternum. For the materials for reconstruction of the chest wall use a polyethylene plate in 2 patients, a Marlex mesh in 5 patients and a Marlex mesh with a methyl methacrylate utilizing the sandwich method in 2 patients. The polyethylene plate proved to be satisfactory for maintaining stability of the chest wall. However, our experience using this plate showed intensive foreign body reactions, and discovering these we have not use the polyethylene plate. Marlex mesh had an affinity with human tissue and was very easy to use and produced no complications. We use Marlex mesh for flat surfaces on the anterior or posterior chest wall. Marlex mesh and methyl methacrylate utilizing the sandwich method, combined the solidity and shaping of methyl methacrylate with the benefit such as easy fixation and excellent incorporation of the mesh.

[Treatment of pericardial cyst under thoracoscopy].

Thoracoscopic therapy was carried out on two cases of pericardial cyst. The first patient was a 24-year-old male. An abnormal shadow on a chest X-ray was pointed out at a regular checkup. Thoracoscopy was carried out under the local anesthesia and a thin-walled cyst was discovered. The cyst was punctured and serous fluid was aspirated. Then, several biopsy specimens were obtained from the cyst wall, a pathological diagnosis of the cyst was made as a pericardial cyst. One year after the thoracoscopy, no abnormal shadow is observed on chest X-ray. The second patient was a 26-year-old male. It was also discovered that he had an abnormal shadow on a chest X-ray at a regular medical checkup. Since a solid mass couldn't be completely denied, thoracoscopy was carried out in preparing for thoracotomy under the general anesthesia. The cyst observed between SVC and the azygos vein, and serous fluid was aspirated form the cyst. Following this, the cyst wall was biopsied and opened. Since no cases of malignant pericardial cysts have been reported, an operation is not usually required for these patients. We suggest that thoracoscopy is very useful tool for the final diagnosis and therapy of pericardial cyst because this method is easily carried out under local anesthesia.

[Mediastinal neurilemmoma originating in the right phrenic nerve: a case report].

We report on a 31-year-old man with a mediastinal neurilemmoma originating in the right phrenic nerve. The patient was admitted because of abnormal chest X-ray shadows observed during a routine checkup. A preoperative diagnosis of bronchial cyst in the mediastinum was made. The tumor was resected by video-assisted thoracic surgery. It was 3 x 4 x 3 cm in size, weighted 15 g, and originated in the right phrenic nerve. The histopathologic diagnosis was mediastinal neurilemmoma. Only 14 cases have been reported in the Japanese literature.

[An evaluation of surgical treatment for giant bullae].

We have experienced 67 operative cases of giant emphysematous bullae of the lung. For the bilateral lesions, two stage operation was performed in five cases and one stage operation under median-sternotomy in 12 cases. Twenty six cases were subjected to Naclerio-Langer's method, 25 cases to partial resection or plication and 15 cases to lobectomy. At the earlier period lobectomy was the treatment of choice, but, recently lobectomy was avoided for the conservation of pulmonary function. In terms of the improvement of pulmonary function, there was no difference between Naclerio-Langer's method and partial resection by Autosuture. In conclusion, for the treatment of giant bullae, partial resection by Autosuture is preferable because of improvement of pulmonary function and one stage median-sternotomy for bilateral lesions also are effective.

Pathological correlations between podocyte injuries and renal functions in canine and feline chronic kidney diseases.

Podocytes cover the glomerulus and their adjacent foot processes form a principal barrier called the slit diaphragm. Podocyte dysfunctions, including podocyte loss and slit diaphragm disruptions, induce chronic kidney diseases (CKD). In this study, we analyzed the correlations between podocyte injuries and renal dysfunctions in domestic carnivores. Dogs and cats were divided into normal and CKD groups according to renal histopathology and plasma creatinine values. Immunostaining results showed that linear reactions of slit diaphragm molecules, e.g., nephrin, podocin, and ACTN4, were parallel to glomerular capillaries in all animals. However, in dogs, reactions of nephrin and ACTN4 were changed to a granular pattern in the CKD group, and their intensities significantly decreased with the number of podocytes in the glomerulus. Moreover, the expression of nephrin and ACTN4 negatively correlated with creatinine. Real-time PCR analysis showed that nephrin mRNA expression in the kidneys of CKD dogs was significantly lower than that in normal animals, and negatively correlated with creatinine. Although no significant correlation between renal dysfunction and podocyte injury was detected in cats, histoplanimetric scores of tubulointerstitial lesions in CKD cats were higher than those in both normal cats and diseased dogs. Furthermore, mRNAs of WT1 and SD molecules were detected in urine from CKD animals. In conclusion, podocyte injuries such as podocytopenia and decreased expression of nephrin and ACTN4 in the glomerulus were more strongly correlated with renal dysfunction in dogs than in cats. These findings suggest that the CKD pathogenesis, especially susceptibilities to podocyte injuries, differed between dogs and cats.

Change in environmental bacterial flora in a new hospital building.

Microbial surveillance of environmental bacteria was performed in order to study the microbial changes in a newly established hospital building. Airborne bacteria and surface-associated bacteria on floors and sinks were systematically collected between 2002 and 2005. The number of isolates obtained from frequently used floors was significantly higher than that obtained from those floors used less often. A significant increase in Staphylococcus aureus, the appearance of Pseudomonas aeruginosa, and changes among species of Gram-negative bacilli were observed 8-11 months after the new building had been opened. Furthermore, pulsed-field gel electrophoresis (PFGE) typing of meticillin-resistant S. aureus (MRSA) and P. aeruginosa showed that strains of the same PFGE groups were isolated from different sinks, floors and the adjoining old buildings. The number of MRSA isolates obtained from the new building increased as time passed. The sinks from which P. aeruginosa strains of the same PFGE type were isolated are connected by the same drainage pipe. Human movement has considerable effects on bacterial flora and their subsequent spread.