Small peptides switch the transcriptional activity of Shavenbaby during Drosophila embryogenesis.

A substantial proportion of eukaryotic transcripts are considered to be noncoding RNAs because they contain only short open reading frames (sORFs). Recent findings suggest, however, that some sORFs encode small bioactive peptides. Here, we show that peptides of 11 to 32 amino acids encoded by the polished rice (pri) sORF gene control epidermal differentiation in Drosophila by modifying the transcription factor Shavenbaby (Svb). Pri peptides trigger the amino-terminal truncation of the Svb protein, which converts Svb from a repressor to an activator. Our results demonstrate that during Drosophila embryogenesis, Pri sORF peptides provide a strict temporal control to the transcriptional program of epidermal morphogenesis.

Minimum incision endoscopic surgery (MIES) in Japanese urology: results of adrenalectomy, radical nephrectomy and radical prostatectomy.

AIM: The aim of this study was to evaluate the feasibility of our minimum incision endoscopic surgeries (MIES), adrenalectomy, radical nephrectomy and radical prostatectomy, which are operated via a single minimum incision which narrowly permits extraction of the specimen, using an endoscope, without gas insufflation, without any trocar ports and without injury to the peritoneum. These operations have been developed in our department in the late 1990 s and have since been performed in more than 1000 patients and certified as advanced surgery by the Japanese government in 2006. METHODS: Adrenalectomy, radical nephrectomy and radical prostatectomy were carried out via a single minimum incision under the conditions of gasless, portless (without trocar ports), intact peritoneum and at low cost with reusable devices. The anatomic plane was separated through the incision and a wide working space was made extraperitoneally. New devices were made especially for this operation in our department, which are now commercially available. The results of the most recent consecutive cases (2005-2007) are evaluated. The results of adrenalectomy and radical nephrectomy performed by 12 operators including inexperienced doctors were compared with the initial results performed by 2 operators, mostly by one operator. RESULTS: In the recent 60 cases of adrenalectomy, the average length of incision, operative time, estimated blood loss were 5.7 cm (5.6 cm in the initial 30 cases), 156 min (147 min) and 174 ml (139 ml), respectively. A complication was observed in one case, injury to the renal artery. Blood transfusion was not performed. Average days to oral feeding, to long walking (more than 100 m) and to possible minimal hospital stay were 1.3 days (2 days), 1.1 days (1.1 days) and 1.9 days (4.6 days), respectively. In the recent 80 cases of radical nephrectomy, the average length of incision, operative time, estimated blood loss were 6.6 cm (6.6 cm in the initial 80 cases), 192 min (186 min) and 315 ml (324 ml), respectively. Complications were not observed in any of the cases (2) and blood transfusion was performed in 1 case (3). Average days to oral feeding, to long walking (more than 100 m) and to possible minimal hospital stay were 1.1 days (1.4 days), 1.2 days (1.4 days) and 1.9 days (4.8 days), respectively. In the recent 50 cases of radical prostatectomy, the average length of incision and operative time were 5.9 cm and 261 min, respectively. Two complications (small rectal injuries) were observed and one blood transfusion was performed. Average days to oral feeding, to long walking (more than 100 m) and possible minimal hospital stay were 1.0 days, 1.0 days and 2.4 days, respectively. Wound pain was minimal and analgesics were generally not required on the second postoperative day in the above 3 operations. Although prophylactic antibiotics were not used in the recent cases of adrenalectomy and radical nephrectomy, surgical site infection was not observed. CONCLUSION: Minimum incision endoscopic surgery (MIES) in Japanese urology is a safe, reproducible, cost-effective and minimally invasive treatment option for adrenal tumor, renal cell carcinoma and prostate carcinoma.

The effect of infliximab on bone metabolism markers in patients with rheumatoid arthritis.

OBJECTIVE: The aim of this study was to evaluate urinary excretion of N-telopeptide of type I collagen (NTX) and deoxypyridinoline (DPD), markers of bone resorption, and serum bone alkaline phosphatase (BAP) level, a marker of bone formation and an early marker of osteoblast differentiation, in patients with rheumatoid arthritis (RA) treated with infliximab. METHODS: Seventeen male and female patients (age 60.7+/-2.53 yr; mean disease duration 12.9+/-3.01 yr; Steinbrocker's class II-IV) with RA, diagnosed according to the criteria of the American College of Rheumatology (ACR), took part in the study between March 2003 and January 2005. None of the patients had a history of oestrogen replacement therapy. All patients were treated with infliximab combined with methotrexate. Infliximab was infused intravenously at 3 mg/kg at baseline, 2 and 6 weeks, then every 8 weeks. To evaluate disease activity, ESR, CRP, the numbers of swollen and tender joints, modified Stanford Health Assessment Questionnaire (mHAQ) score and ACR score were measured. Levels of NTX and DPD in urine and BAP in serum were measured in all patients. RESULTS: ESR, CRP, the number of swollen joints and tender joints, and mHAQ score had decreased significantly 6 weeks after initial treatment and were still low 6 months after initial treatment. NTX levels had decreased significantly 6 weeks after the initial treatment and were still low 6 months after initial treatment. DPD levels had decreased 6 months after initial infusion. Mean serum BAP level did not differ significantly among the three time points. NTX levels were statistically corresponding with the number of swollen joints and mHAQ scores. DPD levels were statistically lower corresponding with ESR. CONCLUSION: Infliximab therapy may inhibit generalized bone loss in patients with RA. NTX is a more sensitive marker than DPD.

Time course of gene expression after the injection of adenoviral vectors containing CTLA4IG gene.

INTRODUCTION: In vivo gene transfection using a recombinant adenoviral vector leads to diminished gene expression in a time-dependent manner that disappears within 4 weeks. CTLA4Ig blocks CD28-mediated costimulatory signal, and inhibits immune responses. We investigated the duration of transgene expression after administration of adenoviral vector containing CTLA4Ig gene (AdCTLA4Ig). METHODS: We injected 1 x 10(9) plaque forming units (pfu) of AdCTLA4Ig into rats (n = 7) via the tail vein. Thereafter, the blood samples were collected for assay of serum CTLA4Ig levels using enzyme-linked immunosorbent assay. RESULTS: The CTLA4Ig level reached the maximum (range, 65-86 microg/mL; average, 75 microg/mL) on days 3 to 5 after injection. Detectable levels of CTLA4Ig were observed up to 49 days. When we injected AdCTLA4Ig in combination with FTY720 administration, the maximum levels were higher and the detectable levels persisted longer. CONCLUSIONS: Because directly injected adenoviral transgene expression had been reported to disappear between 21 to 30 days, we conclude that AdCTLA4Ig inhibits the immune response and prolongs the transgene (CTLA4Ig gene) expression. Some additional immunosuppressants, like FTY720, may be useful to enhance AdCTLA4Ig effects.

[Tension hemopneumothorax complicated by severe hepatic and renal disorder; report of a case].

A 46-year-old man was admitted to our hospital because of dyspnea and chest pain. We diagnosed tension hemopneumothorax and chest tube drainage was performed. A large volume of bloody pleural fluid (1,200 ml) was removed, but severe liver and renal dysfunction were then recognized. He was treated conservatively because there was no more bleeding. Despite administration of methylprednisolone, re-expansion pulmonary edema occurred after 6 hours of drainage, but this was also treated conservatively. After 3 days, his pneumothorax recurrenced. It was successfully managed by video-assisted thoracoscopic surgery (VATS).

[Thoracoscopically diagnosed multicentric Castleman disease; report of a case].

A 57-year-old female was admitted because of chest and back pain. Computed tomography (CT) revealed that many well-marginated lymph nodes were located in mediastinum and abdominal para-aortic area, especially in the right lower mediastinum. These lymph nodes were enhanced at contrast material-enhanced CT. We performed thoracoscopic surgery. The histopathologic diagnosis was multicentric Castleman disease (MCD). MCD should be considered in the differential diagnosis of multiple lymph nodes swelling with hyper globulinemia. Thoracoscopic surgery is the useful method to resect the lymph nodes and diagnose MCD.

[Thoracoscopically resected cystic mediastinal hemangioma; report of a case].

An asymptomatic 66-year-old female was admitted because of an abnormal shadow on chest X-ray. Computed tomography (CT) revealed that a well-marginated round mass with low density, about 3 cm in diameter, was located in the right anterior superior mediastinum. The border was partially enhanced at contrast material-enhanced CT. Magnetic resonance imaging (MRI) [T 2-weighted] showed the lesion as a high intensity tumor. We performed thoracoscopic surgery and resected the easy-bleeding tumor completely. The tumor was dark red in color and contained old blood. The histopathological diagnosis was hemangioma. There was no recurrence for 3 years. Hemangiomas should be considered in the differential diagnosis of well-marginated masses. Thoracoscopic surgery is the very useful methods to resect the mediastinal hemangioma.

Malignant transformation in a mature testicular teratoma left untreated for more than 50 years since childhood.

Although testicular teratoma in childhood is regarded as a benign tumor, little is known about the consequences of pediatric teratoma being left untreated. We report herein a case of malignant transformation observed in a mature testicular teratoma that was presumed to have remained benign for >50 years.

The effect of postoperative ataralgesia by manual therapy after pulmonary resection.

Muscle therapy, a form of manual therapy, was applied to control pain persisting for more than 1 week following posterolateral thoracotomy, and its efficacy for the alleviation of pain was investigated. Eight patients who underwent posterolateral thoracotomy and lung resection for cancer (n=7) or emphysema (n=1) received manual therapy to incised muscles and the muscles inserting into the ribs in the affected area for an average of 17 days postoperatively. Pressure-friction and stretching techniques were used. Treatment was continued until the intensity of the pressure-friction technique reached a level at which the patient complained of pain and a decrease in muscle tone was detected. Treatment was performed once a week for 3 weeks. Pain severity was measured using a visual analog scale (VAS) (0-10). Before the first treatment, the VAS was set at 10, and changes of the score were observed before and after the treatment as well as over time. After three sessions, all patients showed a decrease in pain from 10 to an average of 1.9 (range 1.3-2.6).

Impaired Delta Np63 expression associates with reduced beta-catenin and aggressive phenotypes of urothelial neoplasms.

p63, a homologue of the p53 gene, is considered to be essential for the normal development of stratified epithelia including urothelium. To examine possible roles of p63 in urothelial tumorigenesis, p63 expression was systematically examined in normal urothelium, low-grade papillary noninvasive (LPN) urothelial tumours, and high-grade or invasive carcinomas, using either an isoform-nonspecific or a Delta N-isoform-specific antibody. Expression profiles of p63 were also analysed in cultured cells. Immunoreactivity with the two antibodies was virtually identical in tissue samples examined. Basal and intermediate cell layers of normal urothelium showed intense nuclear p63 immunostaining. This normal staining pattern was preserved in a majority of LPN tumours, whereas it was frequently impaired in high-grade or muscle-invasive carcinomas. At the mRNA level, Delta Np63 expression predominated over TAp63, and amounts of Delta Np63 mRNA correlated with p63 immunoreactivity, confirming that Delta Np63 accounts for p63 expressed in urothelial tissues. In cultured cells, Delta Np63 was also expressed in low-grade tumour cells as well as normal urothelial cells, but undetectable in high-grade aggressive carcinoma cells. Interestingly, impaired Delta Np63 expression significantly associated with reduced beta-catenin expression that was possibly related to progression of urothelial neoplasms. Thus, impaired Delta Np63 expression characterises aggressive phenotypes of urothelial neoplasms.

[Mediastinal signet-ring cell carcinoma of unknown primary site; report of a case].

The question whether the tumor is primary or metastatic sometimes arises in patients with intrathoracic tumor. Especially, adenocarcinoma presents more difficulties in distinguishing primary tumors from secondary or deciding its origin. We reported a case of mediastinal signet-ring cell carcinoma. A 51-year-old female was admitted because of an abnormal shadow during mass screening. Chest X-ray showed a semicircular, well-demarcated shadow on right cardiophrenic angle. Chest computed tomography (CT) revealed a homogeneous mass, approximately 4 cm in diameter. The investigations of whole body failed to reveal any other tumor. At operation, the tumor was adherent to the esophagus but easily resected. Histologically, the tumor was mainly composed of signet-ring cells. Immunohistochemically, surfactant apoprotein (SPA) was strongly demonstrated in tumor cells. SPA is specific to the lung and the tumor was diagnosed as the metastasis in mediastinal lymph nodes. The primary site was considered to exist in lung. But the primary site was not found over 46 months in spite of repeated further examinations.

[Neoadjuvant chemotherapy for invasive bladder cancer].

Although radical cystectomy is the gold standard for invasive bladder cancer, about half of the patients develop distant metastases within two years after operation. Objectives of preoperative (neoadjuvant) chemotherapy in muscle invasive bladder cancer are eradication of micrometastasis and subsequent improvement of prognosis. Based on favorable outcome of M-VAC (methotrexate, vinblastine, adriamycin, cisplatin) therapy in advanced cases, several clinical trials have been performed. Unfortunately, results of the randomized prospective studies failed to demonstrate survival benefits. However, response to the preoperative chemotherapy has been found to be an excellent prognostic factor, and preservation of the bladder (complete preservation or partial cystectomy) is possible in selected cases with complete response. Moreover, good local control by neoadjuvant chemotherapy enables cystectomy in selected cases of locally-advanced cancer once considered unresectable, and may lead to suppression of intraoperative spread of the tumor cells.

Successful treatment of metastatic adenocarcinoma of the urachus: report of 2 cases with more than 10-year survival.

Metastatic urachal cancer is often considered lethal. We report 2 cases of metastatic urachal carcinoma successfully treated with surgical excision followed by combinations of surgery, radiation, and chemotherapy against local recurrence and/or distant metastases, with a recurrence-free survival period of more than 10 years. These cases provide support for multimodal treatments of metastatic urachal cancer.

Suppression of spermatogenesis in ipsilateral and contralateral testicular tissues in patients with seminoma by human chorionic gonadotropin beta subunit.

OBJECTIVES: The pathologic complexity of the testicular tumor makes it difficult to demonstrate exactly the relationship between the impaired spermatogenesis in patients with a testicular tumor and the serum level of the human chorionic gonadotropin beta subunit (beta-hCG). Therefore, we performed quantitative evaluation of spermatogenesis in ipsilateral and contralateral testicular tissues of seminoma to simplify the relation pathologically and endocrinologically and to demonstrate the exact correlation between spermatogenesis and serum beta-hCG levels. METHODS: Fifty-three biopsy specimens from ipsilateral and contralateral testicular tissues of seminoma were analyzed histologically. The quantitative evaluation of spermatogenesis was performed by the mean Johnsen's score count (MJSC). Beta-hCG expression in seminoma was examined immunohistochemically. Serum beta-hCG, testosterone, estradiol, luteinizing hormone, and follicle-stimulating hormone levels were analyzed before orchiectomy. RESULTS: A significant linear relationship (r = -0.82; P <0.005) was found between the serum level of beta-hCG and the MJSC in contralateral testicular tissues but not in ipsilateral ones, although the suppression of spermatogenesis was observed in both sides without suppression of luteinizing hormone and/or follicle-stimulating hormone production. CONCLUSIONS: A clearcut fall in the MJSC with an associated rise in the serum level of beta-hCG was demonstrated in the contralateral testicular tissues but not in the ipsilateral ones of seminoma. It seems most likely that serum beta-hCG suppresses spermatogenesis in both ipsilateral and contralateral testicular tissues without the suppression occurring through the hypothalamus-pituitary-gonadal system, and also that some less well recognized factors affect spermatogenesis, making the relation between serum beta-hCG and MJSC obscure in ipsilateral testicular tissues.

Regulation of vascular endothelial growth factor transcription by endothelial PAS domain protein 1 (EPAS1) and possible involvement of EPAS1 in the angiogenesis of renal cell carcinoma.

BACKGROUND: Endothelial PAS domain protein 1 (EPAS1) is a basic helix-loop-helix/PAS domain transcription factor that expressed most abundantly in highly vascularized organs. The authors examined the effect of transfection of EPAS1 cDNA on the endogenous expression of vascular endothelial growth factor (VEGF) in the 293 Tet-Off cell line and the possible involvement of EPAS1 in the angiogenesis of renal cell carcinoma (RCC). METHODS: Complete cDNA of EPAS1 was cloned and transfected to cells from the 293 Tet-Off fetal kidney cell line, in which the expression of EPAS1 can be inhibited by doxycycline. The subsequent changes in expression pattern of VEGF and transferrin receptor (TfR), a target gene of hypoxia-inducible factor 1alpha (HIF-1alpha), were examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. In addition, expression of EPAS1, HIF-1alpha, and VEGF were analyzed by semiquantitative RT-PCR in five RCC cell lines and in 13 RCC tissue samples. In situ hybridization was performed on 7 of the 13 RCC tissue samples. RESULTS: Endogenous VEGF was increased significantly by the introduction of EPAS1 cDNA at both the mRNA level and the protein level. With the inhibition of EPAS1 by doxycycline treatment, the expression of VEGF was significantly decreased accordingly, whereas the expression of TfR was not affected. EPAS1 was detected in all of the RCC cell lines examined. In RCC tissue samples, EPAS1 mRNA and VEGF mRNA were increased significantly in tumor tissues compared with normal adjacent kidney tissues. In situ hybridization showed that EPAS1 and VEGF were coexpressed topographically in tumor tissues. CONCLUSIONS: These results suggest that endogenous VEGF can be up-regulated transcriptionally by EPAS1, and EPAS1 may be involved in the angiogenesis of RCC.

Inhibitory effect of N-acetylcysteine on invasion and MMP-9 production of T24 human bladder cancer cells.

BACKGROUND: MMPs play a crucial role in the process of cancer invasion and metastasis. METHODS: The influence of NAC on invasion and MMP-9 production of human bladder cancer cell line T24 was investigated using an in vitro invasion assay, gelatin zymography, Western and Northern blot analyses and RT-PCR assays. RESULTS: TPA increased the number of invading T24 cells through reconstituted basement membrane more than 10-fold compared to basal condition. NAC inhibited TPA-enhanced invasion dose-dependently. TPA increased the MMP-9 production by T24 cells without altering expression of TIMP-1 gene, while NAC suppressed TPA-enhanced production of MMP-9. Neither TPA nor NAC altered TIMP-1 mRNA level in T24 cells. In vitro experiments demonstrated that MMP-9 was directly inhibited by NAC but was not influenced by TPA. CONCLUSION: NAC limits invasion of T24 human bladder cancer cells by inhibiting the MMP-9 production in addition to a direct inhibition of MMP-9 activity.

Bone mineral density after total joint arthroplasty of lower extremities in rheumatoid arthritis patients.

We studied the effects on the axial bone mass of total joint arthroplasty (TJA) for lower extremities in 48 female rheumatoid arthritis (RA) patients by using dual-energy X-ray absorptiometry (DXA). Twenty-nine postmenopausal RA patients treated only with nonsteroidal anti-inflammatory drugs (NSAIDs) served as controls. They were studied for an average duration of 63 months. The reduction in the bone mineral density (BMD) of the lumbar spine (L2-4) was significant in both groups (p < 0.01-0.05), but it was not statistically different between the two groups. The BMD of the femoral neck decreased significantly in both groups (p < 0.01-0.05) after 2 years, but it was not statistically different between the two groups. Our data suggest that TJA slowed the rapid axial bone loss usually associated with advanced RA.

Toxic shock syndrome toxin-1 accelerated collagen-induced arthritis in mice.

The aim of this study was to explore the roles of toxic shock syndrome toxin-1 (TSST-1) in collagen-induced arthritis (CIA). DBA/1 mice were immunized with type II collagen (CII) and treated with TSST-1. Intraperitoneal and intravenous injections of TSST-1 aggravated CIA, enhancing its incidence and severity. CIA was accompanied by an increase in anti-CII IgG Ab levels. Intraperitoneal administration with TSST-1 enhanced IFN-gamma, TNF-alpha, and IL-4 production in DBA/1 mice. We discovered the mRNA expressions of IFN-gamma, IL-2, TNF-alpha, IL-1beta, and iNOS in spleen cells stimulated with TSST-1 in vitro. However, IL-12 and IL-4 mRNA expression were seen constitutively without stimulation. Only a little increase of IL-12 and IL-4 mRNA expression was seen at 2-3 h after treatment with TSST-1. Our experiments demonstrated that CIA was aggravated by the treatment with TSST-1, which may have induced various proinflammatory cytokines and the production of both Th1 and Th2 cytokines.