Different origins of two corpora lutea recovered from a pregnant African elephant (Loxodonta africana).

Elephant ovaries contain multiple corpora lutea (CLs) throughout pregnancy. Two CLs (P-1 and P-2) collected from a pregnant African elephant were used to investigate their origin and physiological state in this study. The mRNA expressions of prolactin receptor, CYP11A and inhibin betaB subunit were higher in P-2 than in P-1, while LHCGR and inhibin betaA subunit mRNA were higher in P-1 than in P-2. Protein expression of cleaved caspase-3 was detected in P-1 but not in P-2. These results suggest different origins for the two CLs in this one pregnant elephant, and we also demonstrated the production of bioactive prolactin by the elephant placenta.

Randomised phase II trial of S-1 plus oxaliplatin vs S-1 in patients with gemcitabine-refractory pancreatic cancer.

BACKGROUND: This randomised, open-label, multicenter phase II study compared progression-free survival (PFS) of S-1 plus oxaliplatin (SOX) with that of S-1 alone in patients with gemcitabine-refractory pancreatic cancer. METHODS: Patients with confirmed progressive disease following the first-line treatment with a gemcitabine-based regimen were randomised to receive either S-1 (80/100/120 mg day(-1) based on body surface area (BSA), orally, days 1-28, every 6 weeks) or SOX (S-1 80/100/120 mg day(-1) based on BSA, orally, days 1-14, plus oxaliplatin 100 mg m(-2), intravenously, day 1, every 3 weeks). The primary end point was PFS. RESULTS: Between January 2009 and July 2010, 271 patients were randomly allocated to either S-1 (n=135) or SOX (n=136). Median PFS for S-1 and SOX were 2.8 and 3.0 months, respectively (hazard ratio (HR)=0.84; 95% confidence interval (CI), 0.65-1.08; stratified log-rank test P=0.18). Median overall survival (OS) was 6.9 vs 7.4 months (HR=1.03; 95% CI, 0.79-1.34; stratified log-rank test P=0.82). The response rate (RR) was 11.5% vs 20.9% (P=0.04). The major grade 3/4 toxicities (S-1 and SOX) were neutropenia (11.4% and 8.1%), thrombocytopenia (4.5% and 10.3%) and anorexia (12.9% and 14.7%). CONCLUSIONS: Although SOX showed an advantage in RR, it provided no significant improvement in PFS or OS compared with S-1 alone.

Multimodal endoscopic treatment for delayed severe esophageal stricture caused by incomplete stent removal.

The usefulness of a covered self-expandable metallic stent for benign esophageal stricture and perforation was well established. In case of benign disease, early stent removal was recommended within 6-8 weeks after placement. A case with severe esophageal stricture caused by incomplete stent removal 7 years after stent placement for spontaneous esophageal rupture was reported. Residual stent fragments could be removed by step-by-step multimodal endoscopic treatment, producing satisfactory luminal diameter of the esophagus. In particular, stent trimming with argon plasma coagulation was safe and effective strategy. The endoscopic stent removal is minimally invasive and should be attempted before surgical intervention; however, it is most important to ensure early stent removal before tissue ingrowth or overgrowth can develop.

Preventive effect of tert-butylhydroquinone on scrotal heat-induced damage in mouse testes.

To investigate the effect of tert-butylhydroquinone (tBHQ) on scrotal heat-induced damage in mice testes, 8-week-old mice were divided into 6 groups and administered with or without tBHQ through diet (10 mg/g), intraperitoneal injection (100 mg/kg body weight), or intratestis injection (12.5 mg/kg body weight), respectively. After single scrotal heat exposure (42 °C for 25 min), trunk blood and testes were collected 48 h later. The testes from diet and intraperitoneal tBHQ-treated mice showed more compact interstitial cells and less germ cell loss in the seminiferous epithelium compared with their corresponding non-tBHQ groups. However, intratestis tBHQ treatment showed no marked difference relative to the non-treatment group. In addition, pre-treatment of tBHQ caused lower testosterone concentrations and reduced expression of cytochrome P450 17α-hydroxylase/17,20-lyase (CYP 17) compared to the corresponding non-tBHQ groups. The results indicated that scrotal heat-induced structural damage was partly prevented by pre-treatment of tBHQ, which could be used as an effective antioxidant for preventing scrotal heat-mediated male infertility.

Expression of inhibin/activin subunits in the equine uteri during the early pregnancy.

The establishment of equine pregnancy is a unique and long process during which a series of physical and possibly biochemical interactions are required between the conceptus and uterus. In this study, we investigated the expression pattern of inhibin/activin subunits in the uterus during early pregnancy. The uteri from four adult mares on cyclic day 13 or pregnancy day 25 were obtained. Immunohistochemical experiments suggested that inhibin/activin subunits were immunolocalized in the luminal and glandular epithelium on pregnancy day 25. In addition, the inhibin α and inhibin/activin ßB subunits were not detected, and inhibin/activin ßA subunit was detected, in the luminal and glandular epithelium on cyclic day 13. Real-time polymerase chain reaction and Western blotting results for the inhibin/activin subunits suggested a significant increase in the expression of inhibin/activin subunit ßB and a significant decrease in the expression of inhibin/activin subunit ßA on pregnancy day 25 compared with those on cyclic day 13. Enzyme-linked immunosorbent assays suggested a significant decrease in the concentration of activin A in endometrium extracts from cyclic day 13 to pregnancy day 25. These results suggest that inhibins or activins synthesized in the uterus, as endocrine factors and necessary nutriments, have different expression patterns and may play different, important roles during early embryonic development of the equine.

Characterization of immunoreactive IGF-I pattern during the peri-ovulatory period of the oestrous cycle of thoroughbred mares and its relation to other hormones.

The aim of this study was to characterize ir-IGF-I pattern and its relation to other hormones during the oestrous cycle in mares. Nine non-pregnant non-lactating pluriparous thoroughbred mares were used. The studied mares were examined ultrasonically and bled daily to follow the ovarian changes and the hormonal milieu for a complete Interovulatory interval (IOI). Two (minor and major) follicular waves were characterized per IOI in thoroughbred mares. The largest follicle of the first follicular wave (DF1) was firstly detected at D - 1.75 ± 0.47 with a growth rate of 2.78 ± 0.14mm/day and maximum diameter of 22.45 ± 0.75mm on day 6.65 ± 0.82. The largest follicle of the second follicular wave (DF2) had a growth rate of 2.15 ± 0.29 mm/day, reached a maximum diameter of 42.70 ± 2.63 mm on D 19.25 ± 0.43. Ir-IGF-I increased significantly prior to ovulation and had a similar pattern to oestrogen (r = 0.84, p < 0.05), suggesting that the ovarian follicles are the main source of circulating ir-IGF-I during the oestrous cycle of mares and that ir-IGF-I may be a crucial factor in follicular differentiation and maturation. In conclusion, this study demonstrated that ir-IGF-I is secreted during the oestrous phase of the cycle concomitant with the development of the future ovulatory dominant follicle, and it may act in synergy with other hormones for the selection and differentiation of the dominant follicle.

[The problem of cardiac surgery with neoplasm].

Neoplasm and cardiac diseases are rarely found at the same time. If the standard treatment for both of these diseases is operation, several problems arise;1 stage or 2 stage operation, which operation should be done 1st etc. Especially, if cardiac operation needs cardiopulmonary bypass, neoplasm may grow faster. In such cases, we must decide operative procedure carefully.

[One-stage coronary artery bypass grafting and revascularization in a patient with ischemic heart disease and arteriosclerosis obliterans].

With the overall increase in ischemic heart disease (IHD), cases combining arteriosclerosis obliterans (ASO) of the lower extremities and IHD are on the rise. Therefore, it is necessary to consider methods and timing of operation. These cases have occasionally large collateral pathways from the internal thoracic artery (ITA) to the femoral artery via the epigastric artery. To prevent irreversible ischemia of the lower limbs after harvesting of ITA, we planned to perform a one-stage operation for ASO-combined IHD. Revascularization of the lower extremities was performed 1st. Subsequently coronary artery bypass grafting with ITA was completed. The operative courses were uneventful.

Elective Matsui-Kitamura stent graft repair for descending thoracic aortic aneurysm and chronic type-B aortic dissection.

BACKGROUND: The position of thoracic endovascular aortic repair (TEVAR) compared to open surgery of the thoracic aorta has changed. This study evaluates outcomes after TEVAR performed electively using our original Matsui-Kitamura stent graft (MKSG) to treat descending thoracic aortic aneurysms (dTAA) and chronic type-B aortic dissection (type-B AD), and elucidates the risk factors for postoperative spinal cord ischemia (SCI). METHODS: TEVAR was performed using an MKSG in 66 patients (age: 70.8+/-9.2 years). The underlying etiology was atherosclerotic change in 39 patients, chronic type-B aortic dissection in 23 patients, and other in 4 patients. RESULTS: No perioperative deaths occurred. Three patients showed temporary paralysis due to postoperative SCI. Abdominal aortic aneurysm (AAA) surgery was a risk factor for postoperative SCI (P=0.04). The 5-year survival rate was 81.2%. CONCLUSION: The present study demonstrated that TEVAR of patients with dTAA and chronic type-B AD using an MKSG can be performed with high technical success rates and low rates of severe acute complications. AAA surgery was a risk factor for postoperative SCI.

[Primary anterior mediastinal huge lipoma: report of a case].

We report a case of anterior mediastinal lipoma. A 71-year-old female was admitted for cough. A fat density tumor from anterior mediastinum to left thoracic cavity was found by chest X-ray, chest computed tomography (CT) and magnetic resonance imaging (MRI). Defect of left dorsal diaphragm was suspected and there was a possibility that the tumor was connected to retroperitoneum. Under the preoperative diagnosis of a benign huge mediastinal lipoma, we conducted an operation. At 1st, we observed by thoracoscopy and made sure that the mass was primary anterior mediastinal tumor and not connected to retroperitoneum. Through the median sternotomy, we completely resected the tumor with thymus. The tumor showed 27 cm in diameter, and histopathological diagnosis of the tumor was benign lipoma. Lipoma of the mediastinum is very rare and about 0.3% of all mediastinal tumors. It is sometimes difficult to distinguish huge lipoma from liposarcoma only by clinical examinations such as CT scan or MRI. We evaluated the condition of the tumor by thoracoscopic observation, and the tumor was safely and completely resected by median sternotomy.

Curcumin as a promising antifungal of clinical interest.

OBJECTIVES: The antifungal activity of curcumin was evaluated against 23 fungi strains and its in vitro inhibitory effect on the adhesion of Candida species to human buccal epithelial cells (BEC) was also investigated. METHODS: The antifungal susceptibility was evaluated by broth microdilution assay following the CLSI (formerly the NCCLS) guidelines. The inhibitory effect of curcumin on the cell adhesion was performed with Candida species and BEC. RESULTS: Paracoccidioides brasiliensis isolates were the most susceptible to curcumin while the growth of Aspergillus isolates was not affected. Curcumin was much more efficient than fluconazole in inhibiting the adhesion of Candida species to BEC, particularly those strains isolated from the buccal mucosa of AIDS patients. CONCLUSIONS: The lack of antifungal compounds with reduced side effects highlights the importance of studying natural products for this purpose. Curcumin was a more potent antifungal than fluconazole against P. brasiliensis, the causal agent of the neglected disease paracoccidioidomycosis. Curcumin dramatically inhibited the adhesion of Candida species isolated from AIDS patients to BEC, demonstrating that curcumin is a promising lead compound that warrants further investigation into its therapeutical use in immunocompromised patients.

Protein kinase N (PKN) and PKN-related protein rhophilin as targets of small GTPase Rho.

The Rho guanosine 5'-triphosphatase (GTPase) cycles between the active guanosine triphosphate (GTP)-bound form and the inactive guanosine diphosphate-bound form and regulates cell adhesion and cytokinesis, but how it exerts these actions is unknown. The yeast two-hybrid system was used to clone a complementary DNA for a protein (designated Rhophilin) that specifically bound to GTP-Rho. The Rho-binding domain of this protein has 40 percent identity with a putative regulatory domain of a protein kinase, PKN. PKN itself bound to GTP-Rho and was activated by this binding both in vitro and in vivo. This study indicates that a serine-threonine protein kinase is a Rho effector and presents an amino acid sequence motif for binding to GTP-Rho that may be shared by a family of Rho target proteins.

[Inflammatory myofibroblastic tumor of the lung; report of a case].

A 24-year-old male admitted to our hospital with a pulmonary nodule detected by his chest X-ray and computed tomography (CT). His laboratory findings were within normal limits. Chest CT showed a 10 mm solitary nodule in the right lower lobe. Positron emission tomography showed moderately positive detection correspond to the nodule. We couldn't rule out a malignant tumor and performed partial resection of the right lower lobe. Pathological findings definitely revealed pulmonary inflammatory myofibroblastic tumor. This case was reported together with some reviews of the literature.

[Video-assisted thoracic surgery for clinical stage I lung cancer].

We evaluated our results of video-assisted thoracic surgery (VATS) performed for lung cancer over 8 years. Between April 2000 and October 2008, a total of 409 (60.9%) underwent VATS for lung cancer. Operative procedures as a radical operation were partial resection in 58 patients, segmentectomy in 64 patients, and lobectomy in 229 patients. There was 1 patient with operative death including hospital death due to pulmonary thromboembolism. In a median follow-up period of 21 months, the 5-year cause specific survival rate was 93.7%. According to operative procedures, the 5-year survival rate was 100% in patients underwent partial resection and segmentectomy, and 91.1% in patients underwent lobectomy. According to pathological stages, the 5-year survival rate was 98.8% in 289 patients with stage IA, 69.1% in 34 patients with stage IB, and 68.2% in 14 patients with stage IIIA. In conclusion, VATS lobectomy and VATS intentional limited resection can be performed with low mortality and good prognosis for clinical stage IA lung cancer patients.

Low expression of gamma-glutamyl hydrolase mRNA in primary colorectal cancer with the CpG island methylator phenotype.

The CpG island methylator phenotype (CIMP+) in colorectal cancer (CRC) is defined as concomitant and frequent hypermethylation of CpG islands within gene promoter regions. We previously demonstrated that CIMP+ was associated with elevated concentrations of folate intermediates in tumour tissues. In the present study, we investigated whether CIMP+ was associated with a specific mRNA expression pattern for folate- and nucleotide-metabolising enzymes. An exploratory study was conducted on 114 CRC samples from Australia. mRNA levels for 17 genes involved in folate and nucleotide metabolism were measured by real-time RT-PCR. CIMP+ was determined by real-time methylation-specific PCR and compared to mRNA expression. Candidate genes showing association with CIMP+ were further investigated in a replication cohort of 150 CRC samples from Japan. In the exploratory study, low expression of gamma-glutamyl hydrolase (GGH) was strongly associated with CIMP+ and CIMP+-related clinicopathological and molecular features. Trends for inverse association between GGH expression and the concentration of folate intermediates were also observed. Analysis of the replication cohort confirmed that GGH expression was significantly lower in CIMP+ CRC. Promoter hypermethylation of GGH was observed in only 5.6% (1 out of 18) CIMP+ tumours and could not account for the low expression level of this gene. CIMP+ CRC is associated with low expression of GGH, suggesting involvement of the folate pathway in the development and/or progression of this phenotype. Further studies of folate metabolism in CIMP+ CRC may help to elucidate the aetiology of these tumours and to predict their response to anti-folates and 5-fluorouracil/leucovorin.

[Multiple schwannoma of the intrathoracic vagal nerve; report of a case].

A 67-year-old male referred to our hospital for the close examination of abnormal chest X-ray findings. Chest X-ray films showed abnormal shadows in the right lung field. Chest computed tomography (CT) scan and magnetic resonance imaging (MRI) revealed the 3 tumors which located in superior, middle and posterior mediastinum. The operation underwent under the diagnosis of neurogenic tumors originating from the right intrathoracic vagal nerve. At the operation, the tumors existed the vagal nerve distal to the recurrent laryngeal nerve and they were resected with the transection of vagal nerve. The masses were histologically diagnosed as schwannoma. We reported a rare case of multiple schwannoma arising from the intrathoracic vagal nerve. He had no postoperative complications, and he has been free from recurrence for 4 years postoperatively.

[On/off-pump beating coronary artery bypass grafting after thoracoplasty].

Off-pump coronary artery bypass grafting (CABG) [OPCAB] or on-pump beating CABG (conventional CABG) was performed in 3 post-thoracoplasty patients. Considering their poor respiratory function after thoracoplasty, OPCAB is considered more suitable than conventional CABG with cardio-pulmonary bypass in such cases with severe coronary lesions. However, because the sternum inclines or the pleural cavity may be polluted in these patients, special care is necessary for the operation. In such cases, it may be impossible to bypass to the circumflex artery in OPCAB, and is necessary to consider the use of percutaneous cardiopulmonary support (PCPS), cardio-pulmonary bypass or hybrid therapy before the operation.

[Intractable pneumothorax secondary to pulmonary metastasis of angiosarcoma].

A 68-year-old male suffered from right pneumothorax and was admitted to our hospital. He had a previous history of angiosarcoma of the scalp, and had received local resection and chemoradiotherapy. Chest computed tomography (CT) on admission revealed right pneumothorax and bilateral multiple thin-walled cavities of the lung. We performed partial resection of right lung. Histopathological examination showed a small metastatic lesion around the thin-walled cavities of the lung. Four months after the 1st lung resection, he suffered left pneumothorax. We performed partial resection of the left lung. Ten days after the 2nd lung resection, left pneumothorax recurred. Nine days later, he also developed right pneumothorax. We performed the 3rd operation for right lung. Thoracoscopy demonstrated multiple bullas in right lung and it showed impossibility for radical surgery. Although surgical resection for pneumothorax secondary to metastatic lung cancer is usually efficient, it is very hard to manage the pneumothorax of metastatic angiosarcoma.

Feeding of phytosterols reduced testosterone production by modulating GnRH and GnIH expression in the brain and testes of male Japanese quail (Coturnix coturnix japonica).

Phytosterols (PS), or plant sterols used as cholesterol-lowering agents, have been shown to act as endocrine-disrupting chemicals in some laboratory animals. Moreover, dietary PS efficiently pass through the blood-brain barrier and accumulate in brain cell membranes. We asked whether the accumulation of PS affects reproduction through the hypothalamic-pituitary-gonadal axis. Thirty male quail chicks were randomly divided into 3 groups (control, 80 mg/kg BW, and 800 mg/kg BW), and daily single doses of PS or vehicle were gavaged into the crop sac from 15 to 100 d of age. At the end of the entire period, half of each group was injected intramuscularly with either 10 µg of chicken gonadotropin-releasing hormone 1 (cGnRH-1) or phosphate-buffered saline solution (PBS) as the vehicle. Blood was collected before and 30 min after cGnRH-1 challenge by jugular venipuncture and decapitation, respectively. The results indicated that testosterone concentrations were low (P < 0.05) before (800 mg/kg BW) and after GnRH challenge in PS-treated quails compared with controls (P < 0.001). However, luteinizing hormone (LH) levels were not different among the groups before cGnRH-1 challenge. In addition, PS-gavaged animals failed to manifest increased LH levels after cGnRH-1 injection (P < 0.01). The same trends were observed in pituitary LH levels at 800 mg/kg BW PS after cGnRH-1 injection (P < 0.05). Real-time PCR results revealed that PS (800 mg/kg BW) feeding reduced expression of GnRH-1 in the brain and testes compared to controls. However, gonadotropin-inhibitory hormone (GnIH) expression was significantly elevated before and after GnRH-1 challenges in the brain and testes. Collectively, these results suggest that brain-mediated effects of PS on gonadal function occurs via the induction of GnIH gene expression, and these indirect effects are less potent than direct effects.

Inhibition of cyclin D1 kinase activity is associated with E2F-mediated inhibition of cyclin D1 promoter activity through E2F and Sp1.

Coordinated interactions between cyclin-dependent kinases (Cdks), their target "pocket proteins" (the retinoblastoma protein [pRB], p107, and p130), the pocket protein binding E2F-DP complexes, and the Cdk inhibitors regulate orderly cell cycle progression. The cyclin D1 gene encodes a regulatory subunit of the Cdk holoenzymes, which phosphorylate the tumor suppressor pRB, leading to the release of free E2F-1. Overexpression of E2F-1 can induce apoptosis and may either promote or inhibit cellular proliferation, depending upon the cell type. In these studies overexpression of E2F-1 inhibited cyclin D1-dependent kinase activity, cyclin D1 protein levels, and promoter activity. The DNA binding domain, the pRB pocket binding region, and the amino-terminal Sp1 binding domain of E2F-1 were required for full repression of cyclin D1. Overexpression of pRB activated the cyclin D1 promoter, and a dominant interfering pRB mutant was defective in cyclin D1 promoter activation. Two regions of the cyclin D1 promoter were required for full E2F-1-dependent repression. The region proximal to the transcription initiation site at -127 bound Sp1, Sp3, and Sp4, and the distal region at -143 bound E2F-4-DP-1-p107. In contrast with E2F-1, E2F-4 induced cyclin D1 promoter activity. Differential regulation of the cyclin D1 promoter by E2F-1 and E2F-4 suggests that E2Fs may serve distinguishable functions during cell cycle progression. Inhibition of cyclin D1 abundance by E2F-1 may contribute to an autoregulatory feedback loop to reduce pRB phosphorylation and E2F-1 levels in the cell.