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.

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.

CPEB1 mediates epithelial-to-mesenchyme transition and breast cancer metastasis.

In mouse mammary epithelial cells, cytoplasmic polyadenylation element binding protein 1 (CPEB1) mediates the apical localization of ZO-1 mRNA, which encodes a critical tight junction component. In mice lacking CPEB1 and in cultured cells from which CPEB has been depleted, randomly distributed ZO-1 mRNA leads to the loss of cell polarity. We have investigated whether this diminution of polarity results in an epithelial-to-mesenchyme (EMT) transition and possible increased metastatic potential. Here, we show that CPEB1-depleted mammary epithelial cells alter their gene expression profile in a manner consistent with an EMT and also become motile, which are made particularly robust when cells are treated with transforming growth factor-ß, an enhancer of EMT. CPEB1-depleted mammary cells become metastatic to the lung following injection into mouse fat pads while ectopically expressed CPEB1 prevents metastasis. Surprisingly, CPEB1 depletion causes some EMT/metastasis-related mRNAs to have shorter poly(A) tails while other mRNAs to have longer poly(A) tails. Matrix metalloproteinase 9 (MMP9) mRNA, which encodes a metastasis-promoting factor, undergoes poly(A) lengthening and enhanced translation upon CPEB reduction. Moreover, in human breast cancer cells that become progressively more metastatic, CPEB1 is reduced while MMP9 becomes more abundant. These data suggest that at least in part, CPEB1 regulation of MMP9 mRNA expression mediates metastasis of breast cancer cells.

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.

A case of trichilemmal carcinoma with distant metastases in a kidney transplantation patient.

Transplant recipients receiving immunosuppressants are at a high risk of cancer, especially skin cancer. Trichilemmal carcinoma is comparatively rare compared with other skin cancers. We report here a first case of trichilemmal carcinoma arising in a kidney transplant recipient. A 63-year-old man who had undergone a living donor renal transplantation at the age of 50 years presented with a 15 × 10 mm lesion on his forehead. The pathological diagnosis after resection was trichilemmal carcinoma. Distant metastases involving the lymph nodes, lung, and liver occurred, and the patient died. Given that trichilemmal carcinoma generally has an indolent clinical course and a low metastatic potential, the present case of trichilemmal carcinoma with an aggressive course resulting in distant metastases is rare.

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.

Saliva collection by using filter paper for measuring cortisol levels in dogs.

Four experiments were conducted to evaluate the accuracy and reliability of noninvasive evaluation of cortisol in saliva of dogs. In experiment 1, we measured the cortisol concentration in the filter paper on which 250-µL cortisol solutions had been quantitatively pipetted and in filter papers dipped in cortisol solution. In experiment 2, we collected the blood and saliva of dogs 3 times at 30-min intervals and compared the cortisol concentrations to examine whether the dynamics of cortisol in the blood and saliva are similar. The results of experiments 1 and 2 showed that the cortisol concentration can be quantitatively measured with this method and that the dynamics of cortisol concentration in the plasma and saliva collected by using filter paper are not different (P = 0.14 for experiment 1 and P = 0.51 for experiment 2). In experiment 3, to investigate the factors related to inducing stress in dogs by using the filter-paper method of collecting saliva, we compared the cortisol concentrations at 0 and 30 min after collecting the saliva of pet dogs. The dog owners completed a survey on their dogs, providing basic information and reporting the collection of their dog's saliva. We found that the cortisol concentrations increased significantly in dogs whose owners spent >2 min collecting saliva (P = 0.005), suggesting that prompt collection of saliva is necessary for accurate assessment of cortisol without induction of a stress response. In addition, the cortisol concentrations increased significantly in dogs whose teeth were not regularly brushed (P = 0.04), suggesting that regular teeth brushing mitigates the effect of the collection process on cortisol concentrations in the saliva, with minimal stress to the dogs. In experiment 4, we measured cortisol concentrations in pet dogs accustomed to having their teeth brushed by their owners, before and after interaction with their owners, to assess whether brushing induces stress in dogs. We detected that the cortisol concentrations significantly decreased after human-dog interaction (P = 0.008), suggesting that this method does not induce stress in dogs. Our study indicates that the method of saliva collection by using filter paper is effective in measuring the cortisol concentrations to evaluate stress, although certain steps are required to enhance accuracy.

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.

[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.

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.

Development of a new aortoscope system for the use of endovascular intervention.

"Balloon aortoscopy" is a technique for viewing inner wall of aorta and used in clinics. By this method, endoluminal aortic surface could be clearly monitored, however, during this period, the aortic blood flow is blocked off by the inflated balloon. To solve this clinical problem, we have been developing a prototype aortoscope system without blocking off aortic flow aiming for the use of an assistive technique for endovascular interventions such as stent-graft placement for aortic aneurysm and have been evaluating through in vitro and in vivo tests. The technique introduced for this purpose was the use of intermittent and instantaneous saline jet controlled by a high-speed electromagnetic valve synchronized to heart beat (diastolic phase). In the previous study, we designed an endoscope with two channels (one for saline discharge and the other for forceps insertion), and confirmed the validity of this method by in vitro and in vivo tests. Based on these findings, in this study, we have newly designed a conventional and low price endoscope system aiming for wide clinical use. From the results of in vitro tests using a mock circulation system, it was confirmed that the newly designed system was capable of visualizing a target installed on an inner surface of the mock system suggesting an availability of the system for an aortoscope without blocking off aortic flow.

Placentation in the African elephant (Loxodonta africana). V: the trophoblast secretes placental lactogen.

OBJECTIVES: To determine if elephant placenta secretes a lactogenic hormone which may function as the principal luteotrophin to maintain ovarian luteal function throughout gestation. STUDY DESIGN AND MAIN OUTCOME MEASURES: To label biopsies of endometrium and placenta recovered from African elephant culled professionally throughout gestation with an anti-human prolactin polyclonal antibody in a conventional immunocytochemical staining technique. RESULTS: All trophoblast cells covering the placental villi and forming 'plugs' in the apical endometrial glands stained strongly and precisely with the anti-human prolactin antiserum throughout gestation. CONCLUSIONS: Elephant trophoblast secretes a placental lactogen (elPL) which may stimulate both the development and secretory function of the large accessory corpora lutea of elephant pregnancy and provide the mitogenic stimulus for placental differentiation and development.

[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.

Neurological deficit after simultaneous open abdominal aortic surgery and thoracic aortic endovascular therapy.

We examined a total of 12 cases; 8 cases were identified by searching the literature on Pubmed (excluding case reports published prior to 2000) and 4 cases were ones we personally encountered. We examined age, sex, history of smoking, and preoperative risk factors as preoperative factors, the access route and coverage of the descending aorta as perioperative factors, and complications and survival time as postoperative factors. Mean coverage of the thoracic aorta was 90.8 mm. In terms of perioperative deaths, 8.3% (1 patient) were due to coagulopathy. Perioperative complications occurred in 16.7% of cases (coagulopathy in 1 patient and paralysis in another). No patients experienced complications or underwent additional treatment during a mean follow-up of 22.9 months. This study suggested that simultaneous open abdominal aortic repair and thoracic aortic endovascular therapy is feasible and also involves few postoperative complications. Paraplegia and paralysis tended to occur less frequently than with two-stage surgery, but further study is needed to explain why this is true.

Video-assisted neck surgery for thyroid tumor: gasless lateral approach.

A 27-year-old woman with a thyroid tumor underwent right hemithyroidectomy to rule out malignancies such as follicular carcinoma of thyroid. A subplatysmal skin flap was dissected after making an incision in the anterior aspect of the neck, and the thyroid gland was exposed through the avascular space between the heads of the sternocleidomastoid muscle. After the insertion of the original retractor, video-assisted hemithyroidectomy was performed using ultrasonic surgical devices and conventional endoscopic forceps. Pathological examination of the resected specimen revealed follicular adenoma, and the postoperative course was uneventful. A lateral gasless approach for video-assisted thyroidectomy enables visualization of an adequate operative field and appears to be a safe and effective method.

Immunohistochemical evidence: testicular and scented glandular androgen synthesis in muskrats (Ondatra zibethicus) during the breeding season.

In order to elucidate the relationship between androgens and the function of the muskrat (Ondatra zibethicus) scented glands during the breeding season, we investigated immunolocalization of steroidogenic enzymes P450scc, 3ßHSD and P450c17 in the muskrat testes and scented glands. Nine adult muskrats were obtained in March (n=3), May (n=3) and July (n=3) 2010. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal P450scc, human placental 3ßHSD and porcine testicular P450c17. Histologically, all types of spermatogenic cells including mature-phase spermatozoa in seminiferous tubules were observed in all testes. Glandular cells, interstitial cells, epithelial cells and excretory tubules were identified in scented glands during the breeding season. P450scc, 3ßHSD and P450c17 were only identified in Leydig cells during the breeding season; P450scc and P450c17 were observed in glandular cells of scented glands, however, 3ßHSD was not found in scented glands during the breeding season. These novel findings provide the first evidence showing that scented glands of the muskrats are capable of locally synthesizing androgens and androgens acting via an endocrine, autocrine or paracrine manner may play an important role in scented gland function during the breeding season.

Evaluation of a newly designed endoscope for observing inner wall of large arteries for the use of endovascular intervention.

A prototype endoscope for observing inner wall of large arteries was specially designed and evaluated through in vitro and in vivo tests. The purpose of this endoscope is to visualize the inner wall of large arteries, e.g., an aorta, without blocking off the blood stream aiming for the use of an assistive technique for endovascular interventions such as stent-graft placement for aortic aneurysm. The technique newly introduced for this purpose was the use of intermittent high-pressure saline jet synchronized to heart beat (diastolic phase). In the previous studies using commercially available bronchoscopes, we confirmed the validity of the system utilizing this technique [1, 2]. Based on these findings, in this study, we have specially designed a new endoscope with two channels, one for saline discharge and the other for forceps, and evaluated its performance through in vitro and in vivo tests. From the results of in vitro tests using a mock circulation system, it was confirmed that the newly designed endoscope was capable of visualizing a target installed on an inner surface of the mock system. Also confirmed through in vivo tests using swine was that we could observe bifurcation in descending aorta, e.g., left renal artery, without stopping off the blood stream.

Luteinizing hormone reduction by the male potency herb, Butea superba Roxb.

To determine if Butea superba Roxb., a traditional Thai male potency herb, has androgenic activity in 60-day-old male Wistar rats, we measured its effects on the pituitary-testicular axis and sex organs. Intact and orchidectomized adult male rats were subdivided into five groups (10 rats/group): distilled water, Butea superba (BS)-10, BS-50, BS-250, and testosterone propionate (TP). They received 0, 10, 50, and 250 mg·kg body weight-1·day-1 BS in distilled water by gavage and 6 mg·kg body weight-1·day-1 TP sc, respectively, during the 30-day treatment period. Blood was collected every 15 days and luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were measured. Changes of weight and histological appearance of sex organs were determined at the end of the 30-day treatment and 15-day post-treatment periods. TP treatment reduced serum FSH and LH levels and significantly increased the weight of the seminal vesicles and epididymis, in accordance with histopathological changes, in both intact and orchidectomized rats. No changes in serum testosterone, LH, and FSH levels were observed in any of the intact rats treated with BS, but a significant increase in seminal vesicle weight was observed only in the BS-250 group. Although a significant reduction in serum LH was detected in the BS-50 and BS-250 groups of orchidectomized rats, no significant change in weight or histology of sex organs was observed. Thus, we conclude that B. superba needs endogenous testosterone to work synergistically to stimulate the accessory sex organ of intact animals and can potentially exhibit an LH reduction effect in orchidectomized animals.

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.