Fibroblast growth factor family aberrations in cancers: clinical and molecular characteristics.

Fibroblast growth factor ligands and receptors (FGF and FGFR) play critical roles in tumorigenesis, and several drugs have been developed to target them. We report the biologic correlates of FGF/FGFR abnormalities in diverse malignancies. The medical records of patients with cancers that underwent targeted next generation sequencing (182 or 236 cancer-related genes) were reviewed. The following FGF/FGFR genes were tested: FGF3, 4, 6, 7, 10, 12, 14, 19, 23 and FGFR1, 2, 3, and 4. Of 391 patients, 56 (14.3%) had aberrant FGF (N = 38, all amplifications) and/or FGFR (N = 22 including 5 mutations and one FGFR3-TACC3 fusion). FGF/FGFR aberrations were most frequent in breast cancers (26/81, 32.1%, p = 0.0003). In multivariate analysis, FGF/FGFR abnormalities were independently associated with CCND1/2, RICTOR, ZNF703, RPTOR, AKT2, and CDK8 alterations (all P < 0.02), as well as with an increased median number of alterations (P < 0.0001). FGF3, FGF4, FGF19 and CCND1 were co-amplified in 22 of 391 patients (5.6%, P < 0.0001), most likely because they co-localize on the same chromosomal region (11q13). There was no significant difference in time to metastasis or overall survival when comparing patients harboring FGF/FGFR alterations versus those not. Overall, FGF/FGFR was one of the most frequently aberrant pathways in our population comprising patients with diverse malignancies. These aberrations frequently co-exist with anomalies in a variety of other genes, suggesting that tailored combination therapy may be necessary in these patients.

[Reconstruction of the left common carotid artery for dissective occlusion detected by intra-operative ultrasonography].

We describe a case of Stanford type A acute aortic dissection. Replacement of the ascending aorta and aortic arch was performed using an "arch 1st technique". Following the completion of replacement, hypotension of the left superficial temporal artery pressure was detected. Ultrasonography revealed dissection of the left common carotid artery (LCCA) and compressive occlusion of the true lumen. Reconstruction of the LCCA was performed in the neck. The patient did well after the operation without any neurological abnormalities.

[Anaphylactic shock related to aprotinin induced by anti-aprotinin immunoglobulin G antibody alone; report of a case].

Anaphylactic shock related to aprotinin has been reported to be induced exclusively in the presence of IgE antibody. And the possibility of anaphylactic shock induced by anti-aprotinin IgG antibody alone was controversial. In this paper, we describe the first case of anaphylactic shock induced by aprotinin-specific IgG antibody alone. A 55-year-old man underwent surgical repair of the descending aorta with the use of aprotinin at 2 months after first aprotinin usage. Immediately after initiation of cardiopulmonary bypass with the continuous infusion of aprotinin, clinical symptoms of anaphylactic reaction were found. Postoperative drug lymphocyte stimulation test for aprotinin and aprotinin-specific IgE antibody were negative, but aprotinin-specific IgG antibody was 163 mg/l and positive.

[Arch first technique using handmade branched graft].

We created a method using handmade branched graft to do the aortic arch surgery easier and safer. We made the branched graft using 12 and 8 mm vascular graft. A 77-year-old man with Stanford type A aortic dissection was operated with this method under deep hypothermia. After aortic root manipulation, perfusion of the aortic arch was stopped and selective cerebral perfusion was established. Left subclavian artery (LSCA) was anastomosed to one of the branches. The perfusion of the LSCA was re-started via one of its branches. Respectively, left common carotid artery and brachiocephalic artery reconstruction and reperfusion were performed in a same fashion. After distal anastomosis, anastomosis between the branched graft and main graft was performed consecutively. Postoperative course was uneventful and there was no complication. The treatment of our branched graft was easier than that of ready-made 4-branched graft. We could perform the operation under clear view for its movability with minimal cerebral ischemic time.

Hepatocyte growth factor/scatter factor is implicated in the mode of stromal invasion of uterine squamous cervical cancer.

OBJECTIVE: The purpose of this study was to examine the relationship of hepatocyte growth factor/scatter factor (HGF/SF) to cell motility and invasion in uterine cervical cancer. METHODS: We examined the expression of HGF/SF and its receptor, c-met, in cervical cancer cell lines SKG-IIIa (squamous cell carcinoma) and Hela-S3 (adenocarcinoma) and in stromal cells of the cervical cancer tissue by reverse transcription-polymerase chain reaction. We studied the effect of HGF/SF on invasiveness of SKG-IIIa and Hela-S3 in an invasion model of the modified Boyden chamber method and by electron microscopy. SKG-IIIa cells were also seeded on the thick Matrigel-coated layer to evaluate the invasion patterns in three-dimensional directions. To investigate the mechanism of an inductive effect of HGF/SF on the invasiveness of SKG-IIIa, we examined the effect of HGF/SF on the expression of intercellular adhesion molecule E-cadherin, cell-substrate adhesion molecules CD44, alpha2beta1, and alpha6beta1, and intracellular skeleton fiber actin in SKG-IIIa in cell enzyme-linked immunosorbent assay (ELISA) and immunofluorescence staining. RESULTS: HGF/SF messenger RNA (mRNA) was detected in stromal cells, and c-met mRNA was detected in SKG-IIIa and Hela-S3. Hela-S3 that initially showed weak intercellular contact freely invaded the Matrigel-coated multiporous membrane without the addition of HGF/SF. In contrast, SKG-IIIa that initially showed strong intercellular adhesion could invade the membrane after the addition of HGF/SF. The same results were represented by an addition of HECD-1, an anti-human E-cadherin antibody. In an experiment with cell culture in a thick Matrigel layer, control SKG-IIIa showed a mirror-ball-like invasion pattern, whereas HGF/SF-stimulated SKG-IIIa spread horizontally over the membrane and migrated through the membrane holes, presenting a tentacular invasion pattern. Migration of SKG-IIIa under the membrane was confirmed by scanning and transmission electron microscopy. The addition of HGF/SF in cell ELISA assay decreased the expression of E-cadherin and actin in SKG-IIIa, but it did not change the expression of CD44, alpha2beta1, and alpha6beta1. Immunofluorescence staining revealed that the expression of E-cadherin in cell membrane was disturbed by HGF/SF. CONCLUSIONS: Our data indicate that HGF/SF produced by stromal cells influences the mode of stromal invasion of squamous cervical cancer by selectively decreasing the expression of both E-cadherin and actin.

Radiation therapy following mastectomy for axillary node-positive breast cancer: indication of chest wall irradiation.

This retrospective study was conducted to determine the indication of chest wall irradiation following mastectomy in axillary node-positive breast cancer patients. Between 1982 and 1993, 103 women with axillary node-positive breast cancer received postoperative radiation therapy following mastectomy using the hockey-stick field, which included the ipsilateral supraclavicular fossa and internal mammary nodes, without the chest wall. Ages ranged from 33 to 73 years (median: 47). Thirty-five patients underwent modified radical mastectomy, 48 radical mastectomy, and 20 extended radical mastectomy. Twenty-two patients had 1-3 positive axillary nodes, and 81 had 4 or more positive axillary nodes. The total doses ranged from 42 to 64 Gy (median 54 Gy) with a daily fraction size of 2 Gy. Adjuvant chemotherapy was given to 75 patients, and hormone therapy was administered to 78 patients. The median follow-up time was 121 months (range, 68-191 months) for the 57 surviving patients. The actuarial overall survival rate and the chest wall control rate at 10 years for all patients were 55% and 85%, respectively. Of the 103 patients, 14 developed chest wall recurrence. In the analysis, status of vascular invasion alone had a significant impact on chest wall control. In patients with definite vascular invasion, 2 of 5 (40%) patients with 1 to 3 positive axillary nodes, and 10 of 31 (32%) with 4 or more positive axillary nodes developed chest wall recurrence. In contrast, no patients without definite vascular invasion developed chest wall recurrence. Factors such as age, menopausal status, pathology, tumor location, extent of resection, estrogen receptor status, total dose, chemotherapy, and hormone therapy did not influence the development of chest wall recurrence. Among node-positive breast cancer patients following mastectomy, those with definite vascular invasion should be delivered chest wall irradiation regardless of the number of positive axillary nodes. In contrast, those without definite vascular invasion need not be administered chest wall irradiation.

Reduced levels of connexin43 in cervical dysplasia: inducible expression in a cervical carcinoma cell line decreases neoplastic potential with implications for tumor progression.

Loss of gap junctional intercellular communication (GJIC) has been linked to aberrant proliferation and an enhanced neoplastic phenotype. Many human tumors, including the cervical carcinoma line HeLa, have been reported to be deficient in expression of the gap junction protein connexin43 (Cx43) and GJIC. To determine if this is an early event in carcinogenesis, we utilized immunohistochemistry to screen a series of cervical biopsy samples and demonstrated a major reduction in Cx43 expression in dysplastic regions compared to normal epithelia. To determine whether this loss influences the neoplastic behavior of cervical carcinoma cells, we have constructed HeLa cell lines in which Cx43 expression can be induced in response to doxycycline. This approach allows for the discrimination of Cx43-mediated effects from those due to pre-existing clonal heterogeneity. Cx43 induction in these cells led to assembly of functional junctions but did not alter growth control in vitro as measured by logarithmic growth, saturation density or focus formation when in co-culture with growth-controlled fibroblasts. However, Cx43 induction decreased two indices of neoplasia: it reduced anchorage-independent growth and attenuated the growth rate of tumor xenografts. These results indicate that established HeLa cell lines are unresponsive to Cx43-mediated signals which are thought to mediate growth control of non-transformed cells, however, Cx43 expression can still reduce aspects of the neoplastic phenotype of these cells, indicating that loss of connexin signaling in dysplastic cells may contribute to their neoplastic progression.

Correlation between growth control, neoplastic potential and endogenous connexin43 expression in HeLa cell lines: implications for tumor progression.

A HeLa cell line, obtained from the ATCC, was cloned and found to exhibit a spectrum of in vitro and in vivo growth characteristics as well as variable expression of endogenous connexin43 (Cx43), a widely expressed gap junction protein implicated in growth control. The majority of clones expressed functional Cx43, which contrasted with previous studies reporting that HeLa cells are completely negative for Cx43 mRNA/protein expression. This endogenous Cx43 expression correlated with increased growth control: Cx43-positive clones exhibited a decreased saturation density and a diminished growth capacity when in co-culture with growth-controlled normal cells in constrast to Cx43-negative clones. Endogenous Cx43 expression was negatively correlated with neoplastic potential as evidenced by attenuated anchorage-independent growth and decreased tumorigenicity in immunodeficient mice. Treatment of Cx43-negative cells with 5-aza-2'-deoxycytidine resulted in expression of Cx43, suggesting gene silencing via DNA methylation. These results support the concept of growth control via junctionally transmitted signals and suggest an epigenetic mechanism for tumor cells to circumvent this control during carcinogenesis. Moreover, the heterogeneous nature of this cell line and the ease of connexin43 gene induction suggest caution in the interpretation of results involving gene transfection using noninducible gene expression systems.

[Emergency CABG for patients with LMT lesion treated by mechanical cardiac support].

We retrospectively analysed the perioperative course of 22 patients who underwent emergency CABG for severe left main trunk lesion. The causes of LMT lesion were spontaneous acute myocardial infarction in 14 patients and failed coronary intervention and coronary angiography in 8 patients. IABP was used preoperatively in all patients and additional coronary perfusion catheter were induced for two patients and PCPS was for one patient. Overall incidence in-hospital operative mortality was 22.7% (5/22), four of them were due to cardiac related causes and one of them was due to acute renal failure. Postoperative level of max CPK enzyme was significantly higher in death cases compared to survivors, 6,330 +/- 3,649 versus 1,299 +/- 1,417 IU/dl. We considered that the most important factors in surgical strategy for acute coronary syndrome with LMT lesion were as follows: improvement of hemodynamics with mechanical cardiac support by IABP or/and PCPS, protection of broad myocardial ischemia using coronary perfusion catheter and urgent surgical revascularization by emergency CABG.

The expression patterns of standard and variant CD44 molecules in normal uterine cervix and cervical cancer.

CD44 is a cell-surface glycoprotein postulated to play a role in tumor-cell metastasis. We have examined the expression of the standard CD44 (CD44s), and alternative spliced variants of CD44 containing variant exons v6, v9, and v1O (CD44v6, CD44v9, and CD44v10 respectively) in 9 samples of normal cervix, 6 samples of cervical intraepithelial neoplasia (CIN), and 11 samples with invasive cervical carcinomas. RT/PCR demonstrated the presence of CD44s in all samples of normal cervix and those with invasive carcinomas. CD44v6 was also found in all normal cervical samples and in 9 tissue samples of invasive carcinomas. The results also suggested that some tumor specimens had several higher molecular transcripts containing exon v6 compared to specimens of normal cervix. Immunohistochemistry detected the presence of CD44s and the absence of CD44v10 in both epithelial and stromal cells in all specimens. In contrast, CD44v6 and CD44v9 were stained positive in epithelial cells but were absent in stromal cells. The intensity of CD44v6 and CD44v9 staining was strongest in normal cervical epithelium followed by CIN, invasive squamous cell carcinoma, and adenocarcinoma. In the malignant samples, heterogeneity in staining intensity among different clusters of tumor cells was observed. Furthermore, poorly differentiated and undifferentiated carcinomas from patients having poor prognosis did not stain at all. This study suggests that variant CD44 molecules may serve an important function in the cell contact of cervical epithelial cells, and that cervical epithelium acquires heterogeneity in the expression of CD44 adhesion molecules during carcinogenesis, which may be related to tumor metastasis.

Heparin-like glycosaminoglycans inhibit leukocyte adhesion to endotoxin-activated human vascular endothelial cells under nonstatic conditions.

Since heparin-like glycosaminoglycans (GAGs) have been reported to possess anti-inflammatory effects, we tested effects of heparin-like GAGs on cell adhesion between endothelial cells (ECs) and leukocytes. Effects of heparin-like GAGs on adhesion molecule expression and leukocyte adhesion were analyzed using cultured human umbilical vein ECs stimulated by endotoxin. Expression of adhesion molecules on ECs was unchanged, but adherence of leukocytes to ECs was significantly reduced by heparin-like GAGs under nonstatic conditions. These results suggest that heparin-like GAGs may act as anti-inflammatory agents partly by inhibition of leukocyte adherence to stimulated ECs under nonstatic conditions and may prevent critical conditions caused by endotoxin.

[A case of invasive thymoma with myasthenia gravis and pure red cell aplasia].

The patient was a 42-year-old man whose main complaints were ptosis, diplopia and muscle weakness. Chest X-ray film revealed a right hilar mass and chest CT showed a solid mass at right anterior superior mediastinum. Laboratory studies revealed myasthenia gravis (MG) on EMG and pure red cell aplasia (PRCA) on hematology. An extended thymectomy, including the thymoma, was performed. Thrombocytopenia appeared postoperatively, but it improved when a platelet transfusion was given. Steroid was administered for postoperative adjuvant therapy, and the MG and PRCA were alleviated. This rare case of simultaneous MG and PRCA as complications of a thymoma is reported.

A hepatobiliary study with Tc-99m N-pyridoxyl-5-methyltryptophan (PMT) in a patient with hepatoblastoma.

A patient with hepatoblastoma in whom a hepatobiliary study with Tc-99m PMT demonstrated homogeneous uptake in the tumor is presented. Sonography showed a large solid mass pedunculated from the right hepatic lobe with a "tumor-in-tumor" pattern. It was hypodense relative to normal hepatic parenchyma on CT scanning without contrast material and turned isodense with contrast material. This represented its hypervascularity, which was confirmed by selective celiac arteriography.

[Diagnosis of the pelvic fracture which requires TAE on plain pelvic radiograph].

We reviewed the initial emergency-room plain pelvic radiographs of 75 patients with pelvic fracture. In thirty-six patients transcatheter arterial embolization (TAE) was performed for retroperitoneal hemorrhage associated with pelvic fracture (TAE group) and in 39 patients (non-shock group) TAE was not required. In TAE group, fractures involving iliosacral region and pubic symphysis were more dominant than in non-shock group. Specific soft tissue abnormal shadow of TAE group was not present. It may be impossible to determine the necessity of TAE in pelvic fracture on the basis of plain pelvic radiograph.

[Transcatheter arterial embolization for postpartum hemorrhage].

A case of postpartum hemorrhage controlled by transcatheter arterial embolization is reported. The determination of the exact location of the bleeding vessel in postpartum hemorrhage may be difficult using angiography. Selective embolization is effective but the selection of the artery to embolize (internal iliac versus uterine artery) is still controversial.

Inflammatory bowel diseases and thymus disorder: reactivity of thymocytes with monoclonal antibodies.

Thymectomy is one of the most effective means of treating myasthenia gravis (MG). Formation of lymphoid follicles in the thymus is seen at high rate in ulcerative colitis (UC) as well as MG. We examined the subsets of lymphocytes in the thymuses taken from patients with inflammatory bowel diseases (IBD), using OK series monoclonal antibodies. Decreases in OKT8 (CD8)-, 6(CD1)- and 10-positive cells and an increase of IA1-positive cells were observed in UC. In contrast, there were no significant changes in Crohn's disease (CD). Therefore, the thymocytes from UC appear to consist of comparatively matured cells under the influence of thymic hormone.

[Superselective angiography of lower gastrointestinal bleeding; a report of three case].

Angiography was performed in three patients with lower gastrointestinal bleeding. Extravasation was demonstrated only with superselective catheterization of a branch of superior mesenteric artery. However, this bleeding was not demonstrable with conventional catheterization of superior mesenteric artery. Superselective catheterization is suggested as a useful technique to detect lower gastrointestinal bleeding.