A neutralizing anti-fibroblast growth factor (FGF) 8 monoclonal antibody shows anti-tumor activity against FGF8b-expressing LNCaP xenografts in androgen-dependent and -independent conditions

BACKGROUNDS: Fibroblast growth factor 8-isoform b (FGF8b) has been detected in human clinical sex-organ related cancers including hormone-refractory prostate cancer. There are, however, few relevant experimental models. A murine monoclonal anti-FGF8 antibody, KM1334, has been shown to neutralize FGF8b and inhibit the growth of androgen-dependent mouse mammary SC-3 cells in vitro and in vivo. In the present study, we evaluated the anti-tumor activity of KM1334 against androgen-dependent and -independent progression of FGF8b-expressing human prostate cancer xenografts. METHODS: FGF8b cDNA was transfected into androgen-dependent human prostate cancer cell line LNCaP, and its xenograft tumors were established subcutaneously in SCID mice with or without castration. KM1334 at the dose of 400 microg/head was injected twice weekly. RESULTS: FGF8b-expressing LNCaP cells secreted FGF8b, showed enhanced level of Erk1/2 phosphorylation, and showed more potent growth properties than mock-expressing cells in vitro and in vivo. KM1334 reduced these properties in vitro, inhibited tumorigenecity in vivo (T/C=0.33), and showed anti-tumor activity against established tumors (T/C=0.47) of FGF8b-expressing cells. FGF8b-expressing LNCaP tumors were androgen-dependent. However, they recurred as androgen-independent FGF8b positive tumors after castration. KM1334 also inhibited the growth of established FGF8b-expressing tumors in the androgen-independent states (T/C=0.47). CONCLUSIONS: These results indicate that humanized monoclonal antibodies, conserving the paratope of KM1334, are a promising candidate for therapy of FGF8b-expressing clinical prostate cancers. Follow-up studies using xenograft models with clinical FGF8b-expressing tumors are required to validate these early findings.     

Laparoscopy-assisted stoma closure

PURPOSE: The aim of this study was to describe our improved technique for stoma closure, laparoscopy-assisted stoma closure (LASC). PATIENTS AND METHODS: Eleven (11) children had LASC at our institute during 2005. Their ages at LASC ranged from 4 to 23 months and their body weight ranged from 3.4 to 10.0 kg. Under general anesthesia, a 5-mm trocar was inserted through an infraumbilical, left-lower, or upper quadrant incision, and laparoscopy was used to observe the bowels, the stoma, the line of separation, and any adhesions. Externally, an incision was made around the stoma circumferentially, and a pair of mosquito forceps was inserted into the abdomen along the attachment between the stoma and the abdominal wall where no intra-abdominal adhesions were present, and the tips of the mosquito forceps were used to free the stoma along the proposed line of separation. Electrocautery was used for hemostasis and for completing the separation. After the stoma was taken down, the bowel was anastomosed and the abdomen closed in layers. RESULTS: All stomas were taken down easily without any complications in all cases. The average time for each stomal separation, from incising around the stoma until the stoma was taken down, was 23.1 minutes (range, 17-42). Injury to the abdominal wall musculature was minimal. There was 1 case of postoperative small bowel obstruction and no wound infection or incisional herniation. CONCLUSIONS: Although our experience is limited to only 11 patients, our LASC procedure appears to be an effective option for stoma closure.     

New primary management for appendiceal masses in children: laparoscopic drainage

PURPOSE: The management of appendiceal masses (AM) in children remains controversial. In this study, we evaluated primary laparoscopic drainage (PLD) for efficacy. METHODS: Eleven (11) consecutive cases of AM (mean age, 8.1 +/- 2.8 years) treated between 2000 and 2004 were the subjects for this study. All had PLD on presentation. If the appendix was seen easily seen after PLD, a laparoscopic appendectomy (LA) was also performed. RESULTS: Eight (8) patients underwent PLD alone (LD group) and 3 underwent PLD and LA (LDLA group). In the LD group, the mean operating time was 87.9 +/- 23.2 minutes, oral feeding was commenced after a mean of 2.3 +/- 0.8 days, patients became afebrile within 4.3 +/- 3.1 days, intravenous antibiotics were ceased after 5.3 +/- 3.1 days, C-reactive protein normalized within 13.6 +/- 4.2 days, drains were removed within 4.0 +/- 1.3 days, and hospital stay ranged from 7 to 15 days. There were no intra- or postoperative complications related to the PLD procedure. Interval LA was performed 6.8 +/- 5.8 months after PLD in 6 of 8 LD group patients and was not performed in the remaining 2 owing to parental refusal. In the LDLA group, operating time ranged from 125 to 150 minutes, and oral feeding commenced 4, 5, and 5 days after LA, respectively. One (1) patient developed an adhesive bowel obstruction after LA, which resolved with conservative therapy; in the remaining 2, there were no complications. All 11 patients are well after a mean follow-up period of 3.7 +/- 1.0 years. A histologic examination of the excised appendices showed mild to severe inflammation. CONCLUSIONS: We recommend that PLD be adopted for the primary management of appendiceal masses, as it would appear to be simple, safe, and effective.     

Immunohistochemical demonstration of protein kinase C isozymes in human brain tumors.

Immunohistochemical studies of the expression of protein kinase C isozymes were done in 38 human brain tumors using monoclonal antibodies to three major isozymes: Type I, Type II, and Type III. The brain tumors, with the exception of 3 medulloblastomas and 2 of 6 pituitary adenomas, showed strong immunoreactivity for the Type III isozyme. Astrocytomas, anaplastic astrocytomas, and glioblastomas also showed weak immunostaining for Type II, whereas other tumors lacked this staining. Immunoreactivity for Type I was present, although weak, in some astrocytic gliomas. There was no correlation between the presence of immunoreactivity for protein kinase C isozymes or the intensity of staining for the Type III isozyme and the pathological grade of malignancy. In normal human brain tissue, Type I is localized mainly in neuronal cells, Type II in the neuropil of the cerebral cortex and the molecular and granular layers of the cerebellum, and Type III almost exclusively in astrocytes. The presence of immunoreactivity for the Type III isozyme in varying tumor cells, including those of non-astrocytic tumors and the presence of the Type II and/or Type I isozymes in astrocytic gliomas demonstrate that the expression of protein kinase C isozymes differs between normal and transformed cells.     

A T cell leukemia line produces factor(s) that render rat neutrophils cytotoxic

Proteose-peptone-induced intraperitoneal neutrophils from rats were activated in terms of tumor cytotoxicity by pretreatment with culture supernatants from a human T cell leukemia line, Jurkat (culture sup). Culture sup-treated neutrophils showed cytotoxicity against various tumor cell lines. The cytotoxicity of culture sup-treated neutrophils was dependent on the number of neutrophils and the concentration of culture sup. Cytostasis by activated neutrophils was observed very early in the assay incubation period (within 6 hr), but cytolysis first occurred at 24 hr after the start of incubation. Factor(s) in culture sup responsible for the activation of cytotoxic neutrophils were stable to temperature and pH treatments, and their molecular weight was higher than 10,000. The responsible factor(s) for activation of cytotoxic neutrophils were different from interleukin-2, serum-derived factor, and bacterial lipopolysaccharide.     

Detailed characterization of reactivities of anti-gastric cancer monoclonal antibodies to carbohydrate antigen

Four murine monoclonal antibodies (MoAbs), KM191, KM206, KM230 and KM231, raised against gastric cancer exhibited very similar reactivities to human carcinoma cells, whereas their abilities in probing the antigen shed from the cancer cells were quite different. We found in this study that the four MoAbs reacted immunologically with the same monosialo gangliosides derived from a gastric cancer cell line by two-dimensional high performance thin-layer chromatography. When the reactivities of the MoAbs to a number of purified gangliosides or oligosaccharides were examined, the carbohydrate structure of the antigen was determined as sialyl Lea. KM231 exhibited the highest binding avidity to the oligosaccharide and the ganglioside among the four MoAbs. In a comparative study of KM231 and NS19-9, which is a widely used sialyl Lea-reactive MoAb, KM231 bound to the oligosaccharide with higher affinity and detected the antigen in tissue sections with higher sensitivity. In addition, KM231 could detect a small amount of the antigen ganglioside in human gastric normal and cancerous mucosa and in gastric cancer cell lines by HPTLC-immunostaining.     

Incidence of von Hippel-Lindau Disease in Hemangioblastoma Patients: The University of Tokyo Hospital Experience from 1954–1998

Summary. Summary. Background: Incidence of von Hippel-Lindau disease among hemangioblastomas is important clinical information affecting the management of hemangioblastomas. Studies from Western countries reported 36–40% for the incidence, but no report has been made on the Japanese population. Method: To investigate the incidence in Japan, we retrospectively analyzed all hemangioblastoma patients treated at The University of Tokyo Hospital from 1954 to 1998. By reviewing medical records and imaging studies, von Hippel-Lindau disease was diagnosed clinically following the currently suggested diagnostic criteria. Findings: There were 82 hemangioblastoma patients recorded during the period, and 14 cases (17%) were compatible with von Hippel-Lindau disease. However, when the incidence was calculated for each of the three 15-year periods, which are 1954–1968 (first), 1969–1984 (second), and 1985–1998 (third), the number increased dramatically in the later periods: 2 of 33 (6%) during the first, 4 of 26 (15%) during the second, and 8 of 22 (36%) during the third period. Such increase occurred after the introduction of whole body CT to our institution in 1981, suggesting that improvement of imaging techniques contributed to the sensitivity of diagnosis. In addition, one recent patient with multiple hemangioblastomas was found to harbor germline mutation of the VHL, thereby being diagnosed as von Hippel-Lindau disease on the basis of molecular genetics. Interpretation: The 40% incidence of von Hippel-Lindau disease in hemangioblastomas suggests that extensive screening for von Hippel-Lindau disease associated neoplasms, and probably molecular genetic examination, is indicated for all patients with hemangioblastomas, which should aim for earlier diagnosis and better management of this devastating hereditary disease.     

In vitro activities of oral cephem and telithromycin against clinical isolates of major respiratory pathogens in Japan

The in vitro antibacterial activities of oral cephem antibiotics and ketolide telithromycin against major respiratory pathogens possessing beta-lactam-resistant mutations (within the pbp gene) and/or macrolide-resistant genes (erm and mef) were examined in clinical isolates collected at 66 institutes in all over the Japan between 2002 and 2003. Telithromycin showed the strongest antibacterial activity against methicillinsusceptible Staphylococcus aureus strains with and without macrolide-resistant genes, such as ermA or ermC gene. All the cephem antibiotics showed potent antibacterial activity against Streptococcus pyogenes, with minimum inhibitory concentrations (MICs) of 0.015 mg/L or lower. Cefdinir had a much higher MIC90 against genotypic penicillin-resistant Streptococcus pneumoniae (gPRSP) than cefditoren and cefcapene (8 mg/L cefdinir vs. 1 mg/L cefditoren and cefcapene). The majority of gPRSP harbored either ermB or mefA, and the antibacterial activity of telithromycin against these strains was decreased however some susceptibility was still sustained. Cefditoren exerted the strongest antibacterial activity against beta-lactamase-negative ampicillin-resistant Haemophilus influenzae, with an MIC90 of 0.5 mg/L. These results underline the importance of checking the susceptibility and selecting an appropriate antibiotic against target pathogens.     

Bradykinin improves left ventricular diastolic function under long-term angiotensin-converting enzyme inhibition in heart failure

Both systolic and diastolic cardiac dysfunction coexist in various degrees in the majority of patients with heart failure. Although ACE inhibitors are useful in the treatment of heart failure, the roles of bradykinin in the systolic and diastolic properties of left ventricular function under long-term treatment of ACE inhibitor have not been fully elucidated. We therefore evaluated the changes in left ventricular function, histomorphometry, and the expression of several failing heart related genes, by use of an orally active specific bradykinin type 2 receptor antagonist, FR173657 (0.3 mg/kg per day), with an ACE inhibitor, enalapril (1 mg/kg per day), in dogs with tachycardia-induced heart failure (270 ppm, 22 days) and compared the effects to enalapril alone. Although there were no differences observed in blood pressure, left ventricular dimension, and percentage of fractional shortening, FR173657 significantly increased left ventricular filling pressure (P<0.01), prolonged the time constant of relaxation (P<0.05), and suppressed the expression of endothelial NO synthase and sarcoplasmic reticulum Ca(2+)-ATPase mRNA (P<0.05). FR173657 also upregulated collagen type I and III mRNA (P<0.05) and increased the total amount of cardiac collagen deposits (P<0.05) in left ventricle compared with that in the enalapril-treated group. In conclusion, endogenous bradykinin contributes to the cardioprotective effect of ACE inhibitor, improving left ventricular diastolic dysfunction rather than systolic dysfunction, via modification of NO release and Ca(2+) handling and suppression of collagen accumulation.