Construction of Canine Herpesvirus Vector Expressing Foreign Genes Using a lacZ-TK Gene Cassette as a Double Selectional Marker

An improved method for constructing canine herpesvirus (CHV) recombinants expressing foreign genes by using the lacZ-TK gene cassette as a double selectional marker was developed. A recombinant CHV carrying the lacZ-TK gene at a targeted gene locus was constructed and used as a parental virus for generating new recombinants. The parental virus formed blue plaques and was sensitive to TK-specific drugs, while newly generated recombinants, in which the lacZ-TK gene was replaced with the desired foreign gene, become both resistant to the TK-specific drugs and formed white plaques. Recombinants were isolated by using the combination of drug selection and color selection. This improved method allows construction of recombinant CHV with great ease, because the drug selection can enrich the frequency of recombinant CHV from 0.01–0.1% to 10–80%. This method was employed to construct a recombinant CHV that expressed rabies virus (RV) glycoprotein (G protein). This revised version was published online in July 2006 with corrections to the Cover Date.     

Fine-needle aspiration diagnosis of Kaposi's sarcoma in a developing country

Fine-needle aspiration (FNA) cytology was performed on 15 patients with peripheral lymphadenopathy and/or skin lesions referred to the Department of Pathology of the Hospital Central of Maputo, Maputo, Mozambique. Epitrochlear lymph nodes were the most frequently aspirated site. All aspirates allowed diagnoses of Kaposi's sarcoma (KS). Smears contained loosely cohesive clusters of bland spindle cells, with a radial arrangement and nuclear crush artifacts. These diagnostic clues have not been described in other spindle-cell intranodal lesions that should be considered in differential diagnoses. Taking into consideration the high prevalence of AIDS and limited resources for diagnosis in Africa, FNA cytology appears to be a useful method for the diagnosis of KS in developing countries, reducing the necessity for surgical lymph node excision.     

Effect of stimulated neutrophils from the synovial fluid of patients with rheumatoid arthritis on lymphocytes—A possible role of increased oxygen radicals generated by the neutrophils

Neutrophils from the synovial fluid (SFN) of 10 patients with active rheumatoid arthritis (RA) were investigated to determine the generation of oxygen intermediates (OI) (O2-, H2O2, OH .), chemiluminescence, and lysosomal enzymes (lysozyme and beta-glucuronidase). Lymphocytes from healthy individuals were cocultured at 37 degrees C for 17 hr with SFN from the patients and the number of OKT4+, OKT8+, and OKT3+ cells and the response to mitogens were determined. A markedly increased OI and slightly elevated lysosomal enzyme levels were observed in SFN from patients. Coculture of lymphocytes with SFN resulted in a decreased number of OKT4+ and OKT8+ cells and a greatly reduced response to Con A and mildly diminished response to PHA, while OKT3+ cells were not affected. The simultaneous addition of superoxide dismutase and catalase restored the impairment of monoclonal antibody reaction and lymphocyte responsiveness almost to control levels. It is suggested that the disturbed immunoreactivity of synovial fluid lymphocytes from RA patients may be due to increased OI generated by stimulated neutrophils.     

Histological studies concerning the development of gastric stump carcinoma by endoscopic mucosal biopsies

225 patients who received partial gastrectomies more than five years ago were examined by endoscopy with mucosal biopsies. In the biopsies taken from mucosa close to the anastomosis, hyperplasia of the foveolar epithelium and glandular cysts, findings identical with "gastritis cystica polyposa", were most often seen following Billroth II resection. Cases in which more than 20 years had elapsed since Billroth II resection revealed the highest rate (65.2%) of such mucosal changes. A close relationship between 2 cases (B-II Group) of gastric stump carcinoma detected near the gastro-enteric anastomosis and such mucosal changes is suggested.     

Comparative study of neuroectodermal tumor and Ewing's sarcoma of the bone. Histopathologic, immunohistochemical and ultrastructural features

We carried out a histologic, immunohistochemical and ultrastructural study of 19 tumors, previously classified as Ewing's sarcoma of the bone (ESB). The tumors were categorized histologically into four groups according to the presence or absence and the distribution of Homer-Wright rosettes, together with the immunohistochemical and ultrastructural features: 1) neuroectodermal tumor of bone (NTB) with diffusely dispersed rosettes (4 cases), 2) conventional ESB totally lacking the rosettes (9 cases), 3) intermediate-type tumor with characteristics of both NTB and ESB (4 cases), and 4) spindle cell-type tumor with focal rosettes (2 cases). Ultrastructurally, NTB contained neurotubules and neurosecretory granules, whereas ESB consisted of only undifferentiated cells. Immunoreactivity with antineural antibodies, such as Leu-7, and those against synaptophysin, NSE, neurofilament, and chromogranin, was detected in the majority of NTB, in intermediate- and spindle cell-type tumors and in two of the nine cases of ESB. In view of these findings, we propose that use of the term "neuroectodermal tumor of the bone" should be restricted to tumors with an extensive distribution of rosettes, and that those without unequivocal rosettes should be classified as ESB, although ESB may be the most undifferentiated form of NTB and some overlap may occur between the two tumor types.     

Ten-year experience in the use of double balloon catheter for kidney procurement from non-heart beating donors in cadaveric kidney transplantation

One-hundred-and-twenty patients underwent first cadaveric kidney transplantation from the non-heart beating donors. All of the organs were procured with the use of double balloon catheter for in situ cooling. The mean warm ischemic time and cold ischemic time were 10.7 +/- 17.0 minutes and 18.9 +/- 11.4 hours, respectively. One- and 5-year graft survival rates were 85.0 and 72.7%, respectively. Among 120 recipients, 30 (25%) grafts functioned immediately (immediate function), 82 (68.3%) grafts functioned after varying length of oliguric periods (delayed function) and 8 (6.7%) grafts never functioned (non-function). The mean age of the donors in the group of immediate function (31.5 +/- 16.1 yr) was significantly lower than those of other two groups. The mean warm ischemic time in the group of immediate function (6.0 +/- 11.2 min) was significantly shorter than that of delayed function. However, there was no significant difference in donor hypotensive episode, types of preservation fluid and cold ischemic time between the groups. The conclusion is that the ultimate result of cadaveric kidney transplant from the non-heart beating donors with the use of double lumen catheter is acceptable despite a relatively high incidence of delayed graft function.     

Results of combined therapy of irradiation and intra-arterial one-shot injection of cisplatin for head and neck cancers

Thirty-six previously untreated cases of head and neck cancer were given a combined therapy of irradiation and intra-arterial, one-shot injection of Cisplatin. As a result, the primary lesions in 35 cases were seen to achieve a 66% complete remission (CR) and a 100% overall response (OR). The lymph metastases in twenty-one cases with positive node achieved a 62% CR and a 100% OR. Good, local control of supraglottic and hypopharyngeal cancers were found to be an advantageous factor considered from the standpoint of the "quality of life". The cumulative survival rate, using Kaplan-Meier's method, was 82% for the first year, and 62% for the second year. No severe side effects were observed.     

Extent and duration of isobaric 0.5% bupivacaine spinal anesthesia by using new open-end pencil point spinal needle

Extent and duration of isobaric 0.5% bupivacaine spinal anesthesia by using two spinal needles; a new 25 gauge open-end pencil point spinal needle (needle tip is open as a hole after pulling out the stylet, Doctor Japan Co., Japan) and a 25 gauge Quincke needle (TOP Co., Japan) were studied clinically in 24 patients for elective lower extremity surgery. Patients were randomly assigned into two groups of 12 patients each according to the spinal needle used. Lumbar puncture was performed between L3-4 with the patient in lateral decubitus position. After isobaric 0.5% bupivacaine 4.0 ml was injected in 20 seconds, the patient was placed in supine position, and the onset of anesthesia was assessed by loss of cold sensation (alcohol sponge), sensory block (pinprick) and motor block (modified Bromage's scale) in every 5 minutes until 30 minutes. Recovery of motor block and adverse effects were assessed by a blinded observer. Sensory block at 30 minutes by Quincke needle was Th 8.4 +/- 1.0 and by new open-end pencil point needle it was Th 8.5 +/- 3.2 dermatomes. Two groups did not differ significantly in sensory block and also in motor block. No adverse effect was observed in both groups. These results indicate that this new open-end pencil point spinal needle is very useful for preventing adverse effects of spinal anesthesia.