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We report on the growth of papillary microcarcinoma during active surveillance and before clinical presentation.We conducted a retrospective study of 169 patients with papillary microcarcinoma who were enrolled in active surveillance at our hospital between 2000 and 2004. Patients were followed for a median of 10.1 years using serial ultrasonography (median, 12 examinations), used to calculate the tumor doubling time. To contextualize tumor growth rates during active surveillance, we calculated the hypothetical tumor doubling time before clinical presentation. To resolve the limitations in tumor doubling time, tumor doubling rates were inversely transformed into doubling rates.The doubling rates (per year) during active surveillance (median: 0.0) were0.5, 0.1 to 0.5, -0.1 to 0.1, and-0.1 in 5, 38, 97, and 29 cases, respectively. The proportions of tumors with rather rapid growth, slow growth, stable, and a decrease in size were 3%, 22%, 57%, and 17%, respectively.Tumor growth of papillary microcarcinomas varies from rather rapid growth to a decrease in size during active surveillance.

Artificial diets affect the growth and gut morphology of carnivorous fishes. In this study, we compared a laboratory raised rockfish, Sebastes schlegelii (a carnivore), fed an artificial diet (Lab fish), and the wild-captured rockfish (Wild fish) to confirm the effect of an artificial diet on pyloric caecal morphology and motility. We also established an in vitro method to quantify pyloric caecal sac motility and compared it between Lab and Wild fish. The pyloric caecal length per unit body length, the pyloric caecal weight per unit body weight, and the pyloric caecal weight per length were significantly greater in the Wild fish than in the Lab fish. Additionally, both of the circular and longitudinal muscularis layers were significantly thicker in the Wild fish than in the Lab fish. The pyloric caecal motor frequency of the Wild fish was higher than that in the Lab fish, but the contractile pressure of the pyloric caeca did not change under conventional conditions. These results suggest that the thickness of the tunica muscularis of the pyloric caeca may be involved in its motility, and the pyloric caeca of the Lab fish may not be able to provide functionality in this respect. In conclusion, under artificial rearing conditions, the artificial diet reduced the thickness of the tunica muscularis and reduced the motor frequency of the pyloric caeca in rockfish.

Objective Genetic testing for RET germline mutation can be useful to distinguish whether a patient with medullary thyroid carcinoma (MTC) is genuinely sporadic or hereditary. Conducting a routine preoperative germline RET genetic screening for all patients with MTC has the clinical benefit, i.e., avoidance of unnecessary total thyroidectomy in the selected patients. We sought to clarify the incidence of germline RET mutation carriers in Japanese patients with apparently sporadic MTC and to address the differences in clinicopathological characteristics between true sporadic MTC and hereditary MTC in these patients, all of whom were treated at Kuma Hospital. Methods A total of 134 patients with apparently sporadic MTC who underwent surgery between 1996 and 2014 were enrolled. All patients underwent a germline RET gene mutation analysis preoperatively. Results Germline mutations in RET proto-oncogene were identified in 20 of the 134 (14.9%) apparently sporadic MTC patients. No significant difference in clinicopathological characteristics was observed between the patients with sporadic MTC ( n = 114) and those with hereditary MTC ( n = 20) except for the RET gene carriers’ younger age at diagnosis and presence of multifocal and bilateral lesions. Conclusion Germline RET mutations were identified in 14.9% of Japanese patients with apparently sporadic MTC. No clearly decisive clinicopathological characteristics was observed to distinguish whether an apparently sporadic MTC case was genuinely sporadic or unconsciously hereditary. For the treatment strategy decision, it is advantageous to conduct a routine preoperative germline RET genetic screening for all patients with MTC, even if their MTC is apparently sporadic.

Isolated whole stomachs of farmed Pacific bluefin tuna Thunnus orientalis were externally batch-cultured in cell culture medium at 24 °C for 8 h. This was done in order to confirm that gastric function was maintained during incubation (Exp. 1) so that the effectiveness of the ex vivo technique outlined in the present study in serving as an artificial method of gastric digestion could be determined (Exp. 2). During experiment 1, fresh or cold-stored stomachs (48 h on ice) were intra-gastrically infused with a saline solution (control group) or a skipjack tuna broth stimulant (infusion group). Pepsin-like protease activities within the gastric effluent of each study group were compared. In both control and infusion groups, fresh stomachs demonstrated higher pepsin-like protease activity. Within fresh stomachs, protease activity subsequent to stimulant infusion was significantly greater than it was in the control group. In cold-stored stomachs, however, no difference was observed in the protease activities of control and infusion groups. According to the results, fresh stomachs respond to luminal stimuli. Furthermore, fresh stomachs exhibited motility during incubation while cold-stored stomachs did not. Mucosal layers within fresh stomachs indicated histological normality subsequent to incubation. In experiment 2, both fresh and cold-stored stomachs were filled with a diet consisting of boiled fish paste. After a period of 8 h, dry matter loss of diet was significantly greater within fresh stomachs than it was in cold-stored stomachs. The level of pepsin-like protease activity within fresh stomachs was also higher than levels observed within cold-stored stomachs during the incubation period. These results suggest that fresh stomachs obtained from bluefin tuna maintained functionality ex vivo and that the technique outlined in the present study could be used effectively in the evaluation of bluefin tuna gastric digestion.

The effect of intra-gastric infusion of skipjack tuna (Katsuwonus pelamis) muscle soluble extract and of carnosine, anserine and histidine, on pepsin-like protease activity was studied in the isolated, externally batch-cultured stomach of the rockfish (Sebastes schlegelii). Stomach was isolated from the fish, then intragastrically infused with the above solutions or with a balanced saline solution (control solution) as the stimulant at 0.1 mL/min for 6h. Intra-gastric efflux was collected for measurement of pepsin-like protease activity. Skipjack tuna soluble extract, but not carnosine, anserine or histidine alone caused significant enhancement of pepsin-like protease activity during the infusion. Pepsin-like protease activity from skipjack tuna soluble extract responded immediately after infusion and was kept for 150 min after infusion. Stomach motility at the end of infusion was also observed. These results suggest that isolated stomach can receive mucosal side stimulants in vitro. It is likely that some effective components for stomach digestion in fish exist in skipjack tuna soluble extract.

Background Since papillary thyroid carcinoma (PTC) with tracheal invasion shows a worse prognosis, aggressive surgical treatment including airway resection has been recommended. However, little is known about local recurrence of PTC with minimal tracheal invasion after shaving or laminated resection of the tracheal cartilage. In this study, we investigated the local prognosis of patients who were intra-operatively diagnosed as having minimal tracheal invasion. Methods Of 6,015 patients who underwent initial surgery for PTC between 1987 and 2004, 127 (2.1%) were intra-operatively suspected of having minimal invasion to the trachea and underwent shaving or laminated resection of the tracheal cartilage. These 127 patients were enrolled in this study. Results Of 127 patients, 107 were intra-operatively regarded as having received macroscopically curative treatment (curative group), whereas minimal carcinoma remnants were suspected in the remaining 20 patients (suspicious group). To date, six patients (4.6%) (one patient in the curative group and five in the suspicious group) showed local recurrence at the tracheal edge. The local disease-free survival of patients in the curative group was better than that of those in the suspicious group ( p = 0.0007), although the cause-specific survival of patients in the two groups did not differ. Cut surfaces of resected carcinoma were also pathologically examined for 119 patients and 58 (48.7%) were diagnosed as carcinoma-positive, but the cut surface status was not related to macroscopic curative treatment. Of six patients showing local recurrence, only one died of lung metastasis of carcinoma. Conclusion The incidence of local recurrence of PTC with minimal tracheal invasion is low at 4.6%. Although this event might not be immediately life-threatening for patients, to improve local prognosis, local curative treatment should be pursued. In cases that are suspected of persistent carcinoma remnants, further shaving or laminated dissection should be performed, and if the carcinoma remnant cannot be eliminated, tracheal fenestration should be considered, although such high-risk tumours may be likely to recur regardless of surgical radicality.

Objective Anaplastic thyroid carcinoma arises from differentiated carcinoma and has a very aggressive character. In this study, we investigated the prognosis of patients with anaplastic carcinoma based on UICC stage. Patients and Methods We investigated the prognosis of 75 patients who were diagnosed as having anaplastic carcinoma at Kuma Hospital between 1983 and 2006. Of these patients, 14, 49 and 12 were classified into Stages IVA, IVB, and IVC respectively. Results Stage IVA patients showed a significantly better prognosis than Stage IVB or IVC patients ( p = 0.0017). All patients with Stage IVC died of carcinoma within 1 year regardless of whether locally complete resection was performed. Prognosis of Stage IVB patients who underwent curative surgery did not differ from that of Stage IVA patients, but the prognosis of those who did not undergo surgery or only received palliative surgery was as poor as that of Stage IVC patients. Conclusions Extensive surgical treatment is recommended for Stage IVA patients and palliative care is appropriate for Stage IVC patients. For Stage IVB patients, surgical treatment as a primary therapy is appropriate only when curative resection of the tumour is expected.

Background Although the responsible genes have not yet been identified, it is known that the risk of nonmedullary thyroid carcinoma is elevated in individuals with 1st-degree relatives with nonmedullary thyroid carcinoma. However, it remains controversial whether the biological character of familial nonmedullary carcinoma (FNMTC) differs from that of sporadic carcinoma. In this study, we investigated the prevalence of familial papillary carcinoma and its biological behavior. Methods Between 1987 and 2004, 6,015 patients underwent initial surgical treatment for papillary carcinoma at Kuma Hospital and 273 (4.5%) were classified as having familial carcinoma. We compared the biological characteristics including prognosis between familial and sporadic papillary carcinomas. Results Disease-free survival and cause-specific survival rates of familial carcinoma did not differ from those of sporadic carcinoma. Familial papillary carcinoma showed multicentricity and recurrence to the thyroid more frequently than sporadic carcinoma. There were no differences in other clinicopathological parameters between the 2 groups. Conclusion Prognosis of patients with familial papillary carcinoma did not differ from that of those with sporadic papillary carcinoma. Although routine total thyroidectomy is recommended for familial papillary carcinoma, its therapeutic strategy can otherwise be the same as that for sporadic papillary carcinoma.

The renin–angiotensin system in the kidney plays a critical role in the regulation of renal hemodynamics and sodium handling through the activation of vascular, glomerular and tubular angiotensin II type 1 (AT1) receptor-mediated signaling. We previously cloned a molecule that specifically bound to the AT1 receptor and modulated AT1 receptor signaling in vitro , which we named ATRAP (for AT1 receptor-associated protein). The purpose of this study is to analyze the renal distribution of ATRAP and to examine whether ATRAP is co-expressed with the AT1 receptor in the mouse kidney. We performed in situ hybridization, Western blot analysis, and immunohistochemistry to investigate the expression of ATRAP mRNA and protein in the mouse kidney. The results of Western blot analysis revealed the ATRAP protein to be abundantly expressed in the kidney. Employing in situ hybridization and immunohistochemistry, we found that both ATRAP mRNA and the protein were widely distributed along the renal tubules from Bowman's capsules to the inner medullary collecting ducts. ATRAP mRNA was also detected in the glomeruli, vasculature, and interstitial cells. In all tubular cells, the ATRAP protein colocalized with the AT1 receptor. Finally, we found that the dietary salt depletion significantly decreased the renal expression of ATRAP as well as AT1 receptor. These findings show ATRAP to be abundantly and broadly distributed in nephron segments where the AT1 receptor is expressed. Furthermore, this is the first report demonstrating a substantial colocalization of ATRAP and AT1 receptor in vivo .

Lordosis is correlated with absence or malfunction of the swimbladder. However, swimbladder abnormalities do not completely explain the occurrence of lordosis. We examined whether muscle activities from vigorous tail beat induced by removal of the caudal fin could induce lordotic malformation in juvenile red sea bream. We also attempted to determine the minimal current velocity for inducing lordosis in relation to total length (TL). For this purpose, we employed relative current velocity in terms of “move of water in times of total body length per second (TL s −1 )”. We compared effects of different water current (2 vs. 4 TL s −1 ) for 10 days on bone malformation using juvenile of 25-mm TL. Exposure to 4 TL s −1 current velocity, but not 2 TL s −1 , induced lordosis. Lordotic juvenile red sea bream had a cuneiform centrum mainly at the 15th vertebra. Lordosis was observed in fish with normal swimbladders as well. Thus, swimbladder abnormalities alone cannot completely explain the occurrence of lordosis. We removed part or all of the caudal fins of fish of 40-mm TL and exposed these fish to 2 TL s −1 current for 10 days. Control fish did not show deformations in the vertebrae. Fish with complete removal of the caudal fin swam with vigorous tail beats. However, fish with complete, upper or lower caudal fin removal (36%, 29% or 7%, respectively) displayed lordosis even at the current velocity of 2 TL s −1 . These findings suggested that the muscle activities from excess beat of the tail against excessive current, but not the mechanical action of the caudal fin, induced lordosis.

We studied the metabolism of various oligosaccharides by carp (Cyprinus carpio) hindgut microbes by measuring gas productivity and organic acid production in gut contents using a 50-microl-scale batch culture system. Carp hindgut contents were incubated with 500 microg each of raffinose, lactosucrose, kestose, lactulose, gentiobiose, 4'-galactosyllactose and 6'-galactosyllactose and soybean-, xylo-, and isomalto-oligosaccharides or none (blank culture) at 25 degrees C for 6 h. The time-course of gas release from the culture (Y microl/culture) was expressed as an exponential function of incubation time (t) [Y=A+Bx(1-e(-kt))]; A, B and k are constants). Potential production of gas (A+B) from soybean-oligosaccharide and raffinose was larger than for the other saccharides except for kestose, and blank culture. The rate constant of gas (k) for lactosucrose was larger than that for isomalto- and xylo-oligosaccharide, lactulose, kestose or blank culture. Net production of total SCFA (sum of acetic, propionic and n-butyric acid weights) from cultures with soybean- and isomalto-oligosaccharides, raffinose, gentiobiose and lactosucrose was greater than that from blank culture. These results suggested that soybean-oligosaccharide and raffinose were potentially highly fermentable oligosaccharides for carp hindgut microbes. Chemical structures of oligosaccharides seem to play an important role in the fermentability. It is also likely that oligosaccharide utilization differs between mammals and teleosts.

Expression of neuronal type nitric oxide synthase and renin in the juxtaglomerular apparatus of angiotensin type-1a receptor gene-knockout mice. Angiotensin type-1a (AT1a) receptor gene-knockout ( AT1a -/- ) mice exhibit chronic hypotension and renin overproduction. In the kidneys of AT1a -/- mice, the activity of neuronal type nitric oxide synthase (N-NOS) was histochemically detected by nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase (NADPHd) reaction combined with N-NOS immunohistochemistry. The localization of renin was detected by immunohistochemistry and the results were analyzed morphometrically. The levels of N-NOS and renin mRNA in the renal cortical tissue were determined by reverse transcription-PCR and Northern blot analysis, respectively. In the renal sections from wild-type mice, NADPHd activity and N-NOS immunoreactivity were localized to the discrete region of the macula densa in contact with the parent glomerulus. In contrast, N-NOS-positive macula densa cells were distributed beyond the original location of the macula densa, occasionally extending to the opposite side of the distal tubules. The mean number of N-NOS positive macula densa cells was significantly increased in AT1a -/- mice (186 per 100 glomeruli) compared with wild-type mice (65 per 100 glomeruli). AT1a -/- mice showed 1.4-times higher N-NOS mRNA levels in the renal cortical tissues than wild-type mice. The plasma renin activity was significantly higher in AT1a -/- mice (205.5 ± 26.1 ng/ml/hr) than in wild-type mice (8.0 ± 0.2 ng/ml/hr). The renin-positive areas per glomerulus and renal renin gene expression were 12-times and 2.6-times higher in AT1a -/- mice than in wild-type mice, respectively. These abnormalities, however, were less remarkable in AT1a -/- mice compared with angiotensinogen-knockout mice. When AT1a -/- mice were fed a high-salt diet, the signal intensity of the NADPHd reaction and the number of positively-stained macula densa cells were significantly decreased. The levels of renal cortical N-NOS mRNA were also suppressed by the treatment. Dietary salt loading produced a parallel decrease in plasma renin activity, renal renin-immunoreactive areas, and the levels of renin mRNA without affecting systemic blood pressure. These results provide evidence for the possible involvement of N-NOS at the macula densa in the increased renin production in AT1a -/- mice.

Genetic deficiency of angiotensinogen produces an impaired urine concentrating ability in mice. Angiotensinogen gene-knockout ( Atg −/− ) mice lacking angiotensin II exhibit chronic hypotension. The present study was designed to investigate pathophysiology of Atg −/− mice from the renal functional view. Wild-type ( Atg +/+ ) and Atg −/− mice at 10 weeks of age were housed in metabolic cages for 24-hour urine collection. When provided free access to water, Atg −/− mice showed an increased urine output and a decreased urine osmolality compared with Atg +/+ mice. Urinary excretion and plasma levels of vasopressin were significantly higher in mutant mice than in wild-type mice. On the other hand, urinary excretion of aldosterone in mutant mice was suppressed to the levels under the detection limit of the assay system. The mean plasma aldosterone level of Atg −/− mice was suppressed to 30% of that of Atg +/+ mice. Plasma levels of creatinine, endogenous creatinine clearance, and urinary electrolyte excretion were not different between these mice. In Atg +/+ mice, urine osmolality was markedly increased from 1929 ± 21 to 3314 ± 402 mOsm/kg during water deprivation, whereas this parameter in Atg −/− mice did not change significantly (from 1413 ± 121 to 1590 ± 92 mOsm/kg). Urinary vasopressin excretion increased during water deprivation from 0.24 ± 0.04 and 0.70 ± 0.08 to 0.42 ± 0.06 and 2.31 ± 0.35 ng/mg creatinine in wild-type and mutant mice, respectively. Histologic study revealed interstitial inflammation, and atrophic changes in the tubules and papilla in Atg −/− mice. In conclusion, a genetic deficiency of angiotensinogen produced an impaired urine concentrating ability and tubulointerstitial lesions, indicating the critical role of angiotensinogen in developing normal tubular function and construction.

Fermentability of dietary cellulose, sodium alginate, chitin, α-starch and kaolin (non-fermentable control) by gut microbes was compared in a tilapia, Oreochromis niloticus, fed a diet containing one of the above supplements (150 g · kg−1) for 14 days. The thickness of the tunica muscularis was also compared among fish fed these diets. The concentration of short-chain fatty acids (SCFA) in gut contents was higher in fish fed the α-starch than in fish fed the sodium alginate, chitin, or kaolin diets. Intestinal contents from fish fed the kaolin (control) diet were incubated in batch culture with each of the other supplements. The volume of released gas and production of SCFA for 24 hr was greater with a α-starch as a substrate than in those with the other substrates. The thickness of the tunica muscularis differed among dietary groups but villus height did not. Fish fed chitin or α-starch had a thicker tunica muscularis than those fed cellulose or sodium alginate. These results suggest that this detritivorous fish digests α-starch to produce SCFA in the intestine by microbial fermentation.

The protein product of the retinoblastoma susceptibility gene, RB, is a nuclear phosphoprotein that modulates transcription of genes involved in growth control via interactions with transcription factors. Renin is a rate-limiting enzyme of the renin-angiotensin system that regulates blood pressure and water-electrolyte balance. Renin gene expression is regulated in a tissue-specific and developmentally linked manner. Similarly, the expression of RB is controlled in a differentiation-linked manner. Thus, to investigate whether RB is involved in the regulation of renin gene expression, we examined the effects of RB on transcriptional activity of the mouse renin (Ren-1C) promoter. The Ren-1C promoter contains two transcriptionally important elements; the RU-1 (-224 to -138) and RP-2 (-75 to -47) elements. RB activated the Ren-1C promoter in human embryonic kidney cells. The promoter element responsible for RB-mediated transcriptional regulation was the RP-2 element. The results of DNA-protein binding experiments showed that RB increased nuclear binding activity to the RP-2 element, and site-directed mutation which disrupted binding of nuclear factors to the RP-2 element markedly reduced RB-mediated activation of Ren-1C promoter in human embryonic kidney cells. These results indicate that the RP-2 element plays an important role in RB-mediated transcriptional regulation of Ren-1C promoter activity in human embryonic kidney cells, thereby suggesting an interesting mechanism by which RB may modulate the renin-angiotensin system.

We examined the effects of dietary lactosucrose (20 mg · kg body mass −1 · day −1 ) on body mass gain and the thickness of intestinal tunica muscularis in artificially reared red seabream fed a commercial diet. Lactosucrose did not affect body mass gain. Fish fed lactosucrose had thicker and tougher intestinal tunica muscularis than control fish. In vitro batch culture of intestinal contents of these fish showed that gut fermentation exists in this marine carnivorous teleost. The fermentation can utilize lactosucrose as a substrate, at least in vitro .

Identification and characterization of adenosine A 1 receptor-cAMP system in human glomeruli. Although adenosine is known to affect renal function through stimulating adenosine receptors, little is known about A 1 receptors in human glomeruli. Thus, we attempted to identify the adenosine A 1 receptor-cyclic AMP (cAMP) system in human glomeruli. Normal renal cortical tissues were obtained at nephrectomy of patients with renal cell carcinoma. Glomeruli were isolated using a graded sieving method or dissected manually under a stereomicroscope. Radioligand binding assay using 2-chloro-N-[ 3 H] cyclopentyl adenosine ([ 3 H]CCPA, an A 1 agonist ligand) was performed at 30°C for 90 minutes. Cyclic AMP (cAMP) produced in glomeruli was measured after incubation with different concentrations of N 6 -cyclohexyladenosine (CHA; A 1 agonist) and a phosphodiesterase inhibitor. The specific binding was saturated within 60 minutes and reversible by adding 1 mM of theophylline. Scatchard plot analysis revealed a single class of binding site (K d = 1.78 ± 0.21 nM, B max = 271.7 ± 35.8 fmol/mg protein). The specific binding was inhibited dose-dependently by various agents in an order suggesting A 1 receptor specificity. CHA inhibited the production of cAMP in microdissected human glomeruli. This inhibitory effect was antagonized by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX; A 1 antagonist). This is the first study revealing the presence of the A 1 receptor-cAMP system in human glomeruli using a radioligand binding assay method and by measuring the cAMP production.

Microinjections of N- methyl- d -aspartate (NMDA) into the medial area of the nucleus tractus solitarii (NTS) of the rat led to a decrease in arterial pressure and heart rate. The NMDA receptor antagonist 2-amino-5-phosphonovalerate (AP5) reduced the cardiovascular responses to NMDA. Depressor and bradycardic responses to aortic nerve stimulation were reduced by AP5 but not by a substance P antagonist, injected into the NTS. High K + stimulation caused a calcium-dependent release of glutamate and aspartate from tissues in the area of the NTS. These results provide evidence of NMDA receptor-mediated modulation of the aortic baroreceptor reflex in the rat NTS.

Microinjections of γ-aminobutyric acid (GABA) and glycine, into the medial area of the nucleus tractus solitarii (NTS) of the rat, led to an increase in arterial pressure and heart rate. The GABA receptor antagonist bicuculline and the glycine receptor antagonist strychnine decreased both of these cardiovascular parameters whereas the GABA uptake inhibitor nipecotic acid produced hypertension. High K + stimulation caused a calcium-dependent release of GABA and glycine from tissues in the area of the NTS. Our results suggest that GABA and glycine may modulate the cardiovascular control within the NTS.