Souhei Kitazawa

Immunohistochemical study of epidermal growth factor and transforming growth factor-β in the penetrating type of early gastric cancer

We report that the penetrating type of early gastric cancer (PEN) is a specific type of early gastric cancer and that the poorly differentiated PEN type could be considered an initial lesion of linitis plastica-type cancer. We performed an immunohistochemical study to clarify the role of growth factors (epidermal growth factor [EGF] and transforming growth factor-beta [TGF-beta]) in the PEN type of early gastric cancer. The results indicated that the PEN type of early gastric cancer has a high growth capacity. Moreover, it was suggested that EGF was involved in its specific infiltrative growth and that both EGF and TGF-beta were involved in its specific scirrhous growth. From these findings, it was assumed that the immunohistochemical staining of EGF and TGF-beta in endoscopic biopsy specimens was useful for the diagnosis of the PEN type of gastric cancer and also for the diagnosis of the initial lesion of linitis plastica-type gastric cancer.

In situ DNA-RNA hybridization using in vivo bromodeoxyuridine-labeled DNA probe

SummaryAn in vivo 5′-bromodeoxyuridine (BrdUrd) labeled DNA probe was used for in situ DNA-RNA hybridization. BrdUrd was incorporated into plasmid DNA by inoculating E. coli with Luria-Bertani (LB) culture medium containing 500 mg/L of BrdUrd. After purification of the plasmid DNA, specific probes of the defined DNA fragments, which contained the cloned insert and short stretches of the vector DNA, were generated by restriction endonuclease. The enzymatic digestion pattern of the BrdUrd-labeled plasmid DNA was the same as that of the non-labeled one. BrdUrd was incorporated in 15%–20% of the total DNA, that is, about 80% of the thymidine was replaced by BrdUrd. Picogram amounts of the BrdUrd-labeled DNA probe itself and the target DNA were detectable on nitrocellulose filters in dot-blot spot and hybridization experiments using a peroxidase/diaminobenzidine combination. The BrdUrd-labeled DNA probe was efficiently hybridized with both single stranded DNA on nitrocellulose filters and cellular mRNA in in situ hybridization experiments. Through the reaction with BrdUrd in single stranded tails, hybridized probes were clearly detectable with fluorescent microscopy using a FITC-conjugated monoclonal anti-BrdUrd antibody. The in vivo labeling method did not require nick translation steps or in vitro DNA polymerase reactions. Sensitive, stable and efficient DNA probes were easily obtainable with this method.

In situ hybridization with polymerase chain reaction-derived single-stranded DNA probe and S1 nuclease.

Abstract A rapid and simplified protocol for in situ hybridization (ISH) with polymerase chain reaction (PCR)-derived single-stranded DNA probes and S1 nuclease revealed transcripts of bone matrix proteins on decalcified skeletal bone specimens. Mouse bone tissue was fixed with 4% paraformaldehyde, decalcified with 20% EDTA, and embedded in paraffin. Each pair of primers for reverse transcriptase –PCR was designed to amplify a 280-bp DNA fragment from the coding region of the mature protein of mouse osteonectin (ON) and a 320-bp fragment from the coding region of mouse osteopontin (OP). Initial PCR products were eluted, purified, and reamplified by unidirectional PCR in the presence of the digoxigenin (DIG)-labeled dUTP. ISH was carried out by proteinase K treatment, hybridization, and washing. The unhybridized single-stranded DNA probe was selectively removed by S1 nuclease treatment. Hybridized probes were visualized with the alkaline phosphatase-conjugated anti-DIG antibody. The transcripts of ON and OP were clearly detected on the thin sections of the decalcified bone. Because this protocol does not require cloning or in vitro transcription, reliable and stable ISH can be done in an ordinary laboratory equipped with a thermal cycler.

Intrasellar meningioma mimicking pituitary apoplexy: Case report

BACKGROUND A purely intrasellar meningioma is extremely rare. All the intrasellar meningiomas reviewed in the literature presented with symptoms of hypopituitarism and/or visual disturbance. A case of intrasellar meningioma with a sudden onset of headache and diplopia, mimicking a pituitary apoplexy, is reported. CLINICAL PRESENTATION This 35-year-old man with a sudden onset of headache and double vision was admitted. Neuroimages suggested an intrasellar tumor. Although a pituitary adenoma was suspected from clinical symptoms, a meningioma was compatible with neuroradiologic findings. INTERVENTION Transsphenoidal and frontoorbito-zygomatic approaches were attempted for removal of the tumor. Pituitary apoplexy could not be proved at either operation. The histopathologic diagnosis of this tumor was a fibrous meningioma. CONCLUSION The meningeal tail sign on magnetic resonance images may be a key to diagnosing intrasellar meningiomas.

Prenatal Imaging of Congenital Cerebral Primitive Neuroectodermal Tumor

A case of fetal brain tumor, which appeared after 32 weeks’ gestation, is presented. Prenatal ultrasonography and magnetic resonance imaging demonstrated a large heterogeneous mass in the right supratentorial region and left enlarged ventricle. A male fetus weighing 2,616 g was delivered at 34 weeks’ gestation by cesarean section and died on the 37th day of life due to rapid growth of the tumor. Following autopsy, the pathohistological examination revealed primitive neuroectodermal tumor. Magnetic resonance imaging in the prenatal management of the congenital brain tumor is efficient in evaluating the expansion and margin of the tumor and intratumoral bleeding, which are not demonstrated by ultrasonography.

Conservative management of post-operative peritoneal cysts associated with endometriosis

Objective: To describe the usefulness of fine‐needle aspiration cytology and drainage, followed by long‐acting GnRH‐agonist therapy in the management of post‐operative peritoneal cysts with endometriosis. Methods: In six women who were diagnosed as having post‐operative peritoneal cysts with endometriosis, fine‐needle aspiration cytology and drainage was performed. Thereafter, four patients were treated with long‐acting GnRH‐agonists for 6 months. Two patients refused the treatment. Results: In all patients fine‐needle aspiration yielded specimens which consisted of a population of mesothelial cells. The mean follow‐up time was 4 years (range 3.5–5). The four patients treated with long‐acting GnRH‐agonists show no evidence of recurrence. In two patients who had no additional treatment, the recurrence of the cyst was noted 2 months and 5 months after the drainage. Conclusion: Combination of fine‐needle aspiration cytology and drainage and subsequent long‐acting GnRH‐agonist therapy can be a useful conservative management of post‐operative peritoneal cysts associated with endometriosis.

Bilateral Interstitial Pneumonic Shadows Caused by Perivascular Fibrosis and Extramedullary Megakaryopoiesis of the Lung in a Case of Advanced Agnogenic Myeloid Metaplasia and Myelofibrosis

A 59-year-old man with progressive and advanced agnogenic myeloid metaplasia, also called idiopathic myelofibrosis, had complications showing bilateral interstitial pneumonic shadows. Pathological assessment of transbronchial biopsy revealed pulmonary perivascular fibrosis and infiltration of megakaryocytes. Autopsy 3 months later showed extramedullary megakaryopoiesis and fibrosis in lung, pleura, kidney, liver and spleen. Histopathological analysis for platelet-derived growth factor (PDGF) and PDGF-receptor revealed an abnormally high expression of the PDGF-receptor-β gene in pulmonary fibroblasts. This is the first description of an association between pulmonary fibrosis and PDGF in idiopathic myelofibrosis.

Different origin of leiomyoblastoma by immunohistochemical study

SummaryLeiomyoblastoma has been regarded as a neoplasm of smooth muscle origin. With recent progress in immunohistostaining techniques, many clinicopathological discrepancies have been pointed out about the origin of leiomyoblastoma. It has been claimed that gastrointestinal non-epithelial tumors should be regarded as stromal tumors in order to study their origin. In the present study, we performed various forms of immunohistostaining in seven cases of leiomyoblastoma to determine their origin. One case expressed desmine and muscle specific actin and was considered to be derived from smooth muscle. Four neoplasms expressed S-100 protein (two cases were also NSE positive) and were thought to be derived from the nerve. Two cases were of unknown derivation. These results suggest that the cells of leiomyoblastoma may arise from a primitive to totipotential cell of neural lineages that may anomalously express smooth muscle filaments.

The Specificity of the Penetrating and the Superficial Spreading Types of Early Gastric Cancer—Classification of Early Gastric Cancer Using the Interactive Image Analysis System—

Abstract: Specific types of early gastric cancer were investigated in accordance with the cancer surface area and the degree of penetration by means of quantitative measurements of the surface area of early gastric cancer using the interactive image analysis system.

Monoclonal antibodies to human germ cell tumors from "routine" paraffin-embedded pathological specimens

SummaryWe have established a method for monoclonal antibody (MoAb) preparation from routine paraffin-embedded tissue of human seminoma as an immunogen. Three 40-μm thich sections were deparaffinized and rehydrated. An eight-week-old BALB/c mouse was immunized intraperitoneally with this extract, which showed no detectable protein bands on sodium laurylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Five monoclonal antibodies (MoAbs) with different characteristics were obtained; one reacted with the nucleus, two with the cytoplasm, and two with the cytoplasmic membrane. One of the MoAbs 5G9 reacted with spermatogonia in normal human tissues and with seminoma, embryonal carcinoma and choriocarcinoma in the testicular tumors.