Cell-Based Biohybrid Sensor Device for Chemical Source Direction Estimation.

This paper describes a method to estimate the direction from which the signal molecule reaches the sensor by using living cells. In this context, biohybrid sensors that utilize a sophisticated sensing system of cells can potentially offer high levels of chemical-detection sensitivity and selectivity. However, biohybrid-sensor-based chemical-source-direction estimation has not received research attention because the cellular response to chemicals has not been examined in the context of directional information. In our approach, we fabricated a device that can limit the interface between the cell-laden hydrogel and the chemical solution of interest to enhance the time difference over which the chemical solution reaches the cells. Chemical detection by cells that express specific receptors is reflected as the fluorescence of the calcium indicator within the cells. Our device has eight chambers that each house 3D cell-laden collagen hydrogels facing circularly outward. The device also works as a cover to prevent chemicals from permeating the hydrogel from above. In our study, by observing the time course of the fluorescence emission of each chamber, we were able to successfully estimate the chemical-source direction within an error range of 7-13°. Our results suggest that a combination of microstructure devices embedded with living cells can be used to exploit cell functionalities to yield chemical-source directional information.

Diffusion-weighted MRI as a potential imaging biomarker reflecting the metastatic potential of upper urinary tract cancer.

OBJECTIVE: To evaluate the role of diffusion-weighted MRI (DW-MRI) as an imaging biomarker for upper urinary tract cancer (UUTC) that has already metastasized or will metastasize soon. METHODS: 61 patients clinically diagnosed with UUTC were prospectively enrolled in this study. All the patients underwent MRI, including DW-MRI, prior to any interventions. Correlations between apparent diffusion coefficient (ADC) and other clinicopathological variables, including metastasis-free survival, were analysed. RESULTS: Median follow-up period was 938 days. Of the 61 patients, 12 had any metastases at the initial diagnosis. 11 patients developed metastases during the follow-up period. These 23 patients were categorized as "Metastatic". Of the remaining 38 patients, 35 with a follow-up period longer than 400 days were categorized as "Localized". ADC was significantly lower in the Metastatic category than in the Localized (p = 0.0002) category. Multivariate analysis of pre-operative variables identified ADC (cut-off value, 1.08 × 10(-3) mm(2) s(-1)) and clinical T stage based on T2 weighted MRI as an independent predictive factor of metastatic UUTC. 46 patients without any metastases during the initial diagnosis were stratified into a high-risk group (16 patients with low ADC and clinical T3-4) and a low-risk group (30 patients with high ADC or clinical Ta-2). The 3-year metastasis-free survivals were 45% and 93%, respectively. CONCLUSION: In the current study, UUTC with lower ADC value is more likely to have metastatic potential. Incorporating ADC with clinical T stage helps to differentiate metastatic UUTC at the initial diagnosis. ADVANCES IN KNOWLEDGE: DW-MRI is a potential imaging biomarker reflecting metastatic propensity of UUTC.

The SOCS and STAT from JAK/STAT signaling pathway of kuruma shrimp Marsupenaeus japonicus: molecular cloning, characterization and expression analysis.

Signal transducer and activators of transcription (STAT) gene, suppressors of cytokine signaling (SOCS) has been isolated from kuruma shrimp, Marsupenaeus japonicus and characterized. The kuruma shrimp STAT (MjSTAT) cDNA was composed of 2901 bp consisting of 801 amino acid residues which includes a protein interaction domain, all alpha domain, DNA binding domain and SH2 domain. Homology analysis of MjSTAT showed 94.1% and 34.0% identities with Penaeus monodon STAT (PmSTAT) and Drosophila melanogaster STAT92E (DmSTAT), respectively. The kuruma shrimp SOCS (MjSOCS) cDNA was composed of 1675 bp consisting of 342 amino acid residues including a SH2 domain and C-terminal SOCS domain. Homology analysis of MjSOCS showed 52.6% and 21.3% identities with Chinese mitten crab (Eriocheir sinensis) SOCS2 and fruit fly (D. melanogaster) SOCS44A, respectively. The MjSTAT and MjSOCS genes are constitutively expressed in the muscle, stomach, brain and gill of kuruma shrimp. In lymphoid organ cells, an enhanced expression of both MjSTAT and MjSOCS genes are observed following stimulation with peptidoglycan and polycytidylic acid. These observations suggest that MjSTAT and MjSOCS might play a major role in the innate immune defense of kuruma shrimp. The discovery of JAK/STAT signaling pathway in shrimp will allow a complete and concrete understanding of shrimp cytokine signaling.

Development of a nomogram incorporating serum C-reactive protein level to predict overall survival of patients with advanced urothelial carcinoma and its evaluation by decision curve analysis.

BACKGROUND: The purpose of this study is to investigate the prognostic impact of C-reactive protein (CRP) on patients with advanced urothelial carcinoma and to develop a novel nomogram predicting survival. METHODS: A total of 223 consecutive patients were treated at Tokyo Medical and Dental Hospital. A nomogram incorporating V was developed based on the result of a Cox proportional hazards model. Its efficacy and clinical usefulness was evaluated by concordance index (c-index) and decision curve analysis. RESULTS: Of the 223 patients, 184 (83%) died of cancer. Median follow-up periods of patients who died and those who remained alive were 5 and 11 months, respectively. We developed a novel nomogram incorporating Eastern Cooperative Oncology Group Performance Status, presence of visceral metastasis, haemoglobin and age. The c-index of the nomogram predicting survival probability 6 and 12 months after diagnosis was 0.788 and 0.765, respectively. Decision curve analyses revealed that the novel nomogram incorporating CRP had a superior net benefit than that without CRP for most of the examined probabilities. CONCLUSION: We demonstrated the prognostic impact of CRP that improved the predictive accuracy of a nomogram for survival probability in patients with advanced urothelial carcinoma.

Indocyanine green angiography for intra-operative assessment in vascular surgery.

OBJECTIVES: Indocyanine green (ICG) angiography is used for the intra-operative assessment of the graft vessel in coronary artery bypass grafting to enable immediate revision if necessary. We report the feasibility and implications of an ICG colour imaging system, HyperEye Medical System (HEMS), in surgeries for arteriosclerosis obliterans (ASO) and abdominal aortic aneurysm (AAA) which carry risk of mesenteric ischaemia. METHODS: HEMS ICG angiography was used for the intra-operative assessment of 12 ASO patients and 10 AAA patients. RESULTS: In the ASO patients, HEMS angiography enabled visualisation of the graft and native artery. The fluorescent lucent region in the artery distal to the anastomosis was shown in 1 of 12 ASO patients. There was a 3-s time lag in the increase of intensity between the proximal artery and distal stenotic region. In AAA patients, HEMS angiography clearly showed the perfusion in the mesenteric arteries and intestinal wall as opaque. One AAA patient had segmental ischaemia due to thromboembolism and another one had diffuse ischaemia due to systemic malperfusion. The ischaemic region of the intestine was visualised as a fluorescent lucent area by HEMS angiography. CONCLUSION: HEMS angiography can accurately assess peripheral arterial perfusion in surgical cases with ASO and AAA.

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.

Granulomatous mycosis fungoides presenting as poikiloderma.

Granulomatous mycosis fungoides (MF) is a rare subtype of MF, characterized by the histological presence of a granulomatous reaction, but distinct clinical characteristics are not present. A 41-year-old healthy man presented with poikiloderma, ichthyosis and erythematous scaly plaque. Histological examination of a biopsy taken from poikilodermic skin showed a granulomatous reaction to epidermotropic atypical lymphocytes. However, in other areas there were only findings of conventional MF without granuloma. Granulomatous MF may be associated with poikiloderma.

Mesothelial cells from tunica vaginalis, a practical source for mesothelial transplantation.

Transplantation of mesothelial cells is used to repair peritoneum that is damaged by surgery, peritonitis, and peritoneal dialysis. The largest obstacle for clinical application of mesothelial cell transplantation is the lack of a reliable source of mesothelial cells. So far, they are isolated from omentum, mesentery, parietal wall and ascites. Procedures used to obtain mesothelial cells from the omentum or mesentery are invasive, however, especially in pre-operative situations. Sufficient amounts of ascites for aspiration can not be obtained under physiological conditions. We have developed a novel method of isolating mesothelial cells from the tunica vaginalis. The tunica vaginalis originates from the peritoneum and descends into the scrotum along with the testis during fetal development. This region provides a source of mesothelial cells that is convenient to approach and free from abdominal complications. Transplantation of autologous mesothelial cells that were isolated from tunica vaginalis was effective in preventing post-operative adhesions. In this review, we summarize mesothelial cell transplantation trials and describe the method of isolating mesothelial cells form the tunica vaginalis. Mesothelial cell transplantation might be widely accepted for clinical use in the near future.

[Conformation of the ribosomal protein S1 of Thermus thermophilus in solution under different ionic conditions].

The structure of protein SI of Thermus thermophilus (M = 61 kDa) in solution at low and moderate ionic strengths (0 M and 100 mM NaCl, respectively) has been studied by small-angle X-ray and neutron scattering. It was found that protein S1 has a globular conformation under both ionic conditions. The modelling of different packing of six homologous domains of S1 on the basis of the NMR-resolved structure of one domain showed that the best fit of calculated scattering patterns from such complexes to experimental ones is observed at a compact package of the domains. The calculated value of the radius of gyration of the models is 28-29 angtroms, which is characteristic for globular proteins with a molecular mass of about 60 kDa. It was found that protein S1 has a tendency to form associates, and the type of the associate depends on ionic strength. These associates have, in general, two or three monomers at a moderate ionic strength, while at a low ionic strength the number of monomers exceeds three and they are packed in a compact manner. Strongly elongated associates were observed in neutron experiments at a moderate ionic strength in heavy water. The association of protein molecules was also confirmed by the data of dynamic light scattering. From these data, the translational diffusion coefficient of protein S1 at a moderate ionic strength was calculated to be (D20,w = (2.7 +/- 0.1) x 10(-7)cm2/s). This value is essentially smaller than the expected value (D20,w = (5.8 - 6.0) x 10(-7)cm2/s) for the S1 monomer in the globular conformation, indicating the association of protein molecules under equilibrium conditions.

Long-term outcome of bladder papillary urothelial neoplasms of low malignant potential.

OBJECTIVE: To evaluate the long-term outcome of bladder papillary urothelial neoplasms of low malignant potential (PUNLMP). PATIENTS AND METHODS: Of 475 consecutive patients with newly diagnosed bladder tumours between 1976 and 1993, 330 (69%) had superficial (Ta and T1) tumours and 53 (11%) were diagnosed as having PUNLMP. Fifty patients (mean age at presentation 57.2 years, range 26-83; male-to-female ratio 6 : 1) who were followed for> 5 years or until they died, were included in the present study. All histological slides were reviewed, and fulfilled the diagnostic criteria of the 1998 World Health Organization/International Society of Urological Pathology classification system. RESULTS: The mean (median, range) follow-up was 11.7 (10.8, 1.3-24.4) years. During the follow-up, 30 patients (60%) had local recurrences. The 2, 5 and 10-year recurrence-free rates were 66%, 51% and 36%, respectively. No patients developed high-grade or muscle-invasive (>/= T2) carcinomas, or upper urinary tract tumours, or died from the disease. At the last follow-up, 34 patients (68%) had been disease-free for> 5 years. CONCLUSIONS: Despite a high recurrence rate, PUNLMP carries a very low malignant potential. We agree with the use of the term 'papillary urothelial neoplasms of low malignant potential' instead of 'superficial bladder carcinoma (cancer)' for these tumours.

Spontaneous regeneration of the seriously injured sympathetic pathway projecting to the prostate over a long period in the dog.

OBJECTIVES: To explore the spontaneous regeneration, over a long period, of the seriously injured sympathetic pathway controlling the prostate. MATERIALS AND METHODS: The hypogastric nerve (HGN), which is part of the sympathetic pathway from the spinal cord to the prostate, was partly removed over half of its length on both sides in six dogs. Four years after surgery the responses of the prostate to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN proximal to the site removed, were assessed. RESULTS: In six dogs, 10 of the 17 LSNs (second to fourth) and four of the 10 HGNs stimulated elicited prostatic contraction. The pathways via the ipsilateral HGN and/or the contralateral HGN from the LSNs to the prostate were identified as having regenerated in four of six dogs. CONCLUSION: These results indicate that the sympathetic pathways via the HGN to the canine prostate can regenerate spontaneously over a long period after serious injury, and that their cross-innervation system can also be repaired.

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.

Regeneration of hypogastric nerve using a polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge proved electrophysiologically in a canine model.

The hypogastric nerve (HGN) is a sympathetic nerve in the peritoneal cavity and controls urinary and seminal functions. In this study, the regeneration of HGN was determined by using a new type of an artificial nerve conduit, polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge in two dogs. A PGA-collagen nerve conduit (diameter=2 mm) was interposed in a 10 mm gap of the right HGN. The regeneration of the HGN was evaluated electrophysiologically 8 months after the operation. The intraluminal pressure of spermatic duct and the bladder neck were elevated 80 mmHg and 25 mmHg respectively by the stimulation across the regenerated HGN. The prostate contraction was also elicited. The responses diminished after the excision of the regenerated portion of HGN. These results proved the regeneration of HGN and this nerve conduit will be great help for patients who suffer from urinary and seminal disturbances.

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.

Detection of matrix metalloproteinases in canine mammary tumours: analysis by immunohistochemistry and zymography.

Expression of matrix metalloproteinases (MMPs) was examined immunohistochemically (MMP-9 and -2) and by gelatin zymography (MMP-9) in 12 benign and 12 malignant canine mammary tumours. In nine of 12 benign tumours, weak expression of MMP-9 was demonstrated immunohistochemically in the cytoplasm of neoplastic cells. In the remaining three benign tumours (mixed or complex) MMP-9 expression was observed in the neoplastic luminal epithelial cells and myoepithelial cells in some areas. The neoplastic luminal epithelial cells in 12 malignant tumours reacted strongly for MMP-9. MMP-2 immunolabelling was observed in both benign and malignant tumour tissues, but was stronger in the latter. Zymography and densitometry showed that expression of MMP-9 was higher in the benign mammary tumours than in normal canine mammary tissues, but highest in the malignant mammary tumours. In benign and malignant canine mammary tumours, the zymography and densitometry results for MMP-9 accorded well with the immunohistochemical results obtained with anti-human MMP-9 monoclonal antibody.

Eimeria organisms develop in the epithelial cells of equine small intestine.

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.

Involvement of apoptosis in the control of Sertoli and pre-meiotic germ cell numbers in the developing rabbit testis.

The numbers of Sertoli and pre-meiotic germ cells in the developing rabbit testis were investigated as an initial step in determining the physiological meaning of the control of cell number in the testis by apoptosis. Sections were stained immunohistochemically for the detection of apoptotic cells and counterstained with haematoxylin. The number resulting from subtraction of the number of apoptotic cells from the total cell number was defined as the viable cell number. The number of viable premeiotic germ cells in the adluminal compartment of seminiferous tubules decreased during the pre-natal period, although neither apoptotic nor necrotic figures were detected. After birth, the numbers of total and apoptotic Sertoli and pre-meiotic germ cells increased, maintaining a stable ratio of their viable cell populations until the induction of meiosis. During induction of meiosis, the increase in the number of viable Sertoli cells was significantly accelerated because of the rapid decrease in the number of apoptotic Sertoli cells. Just after spermatids were formed the number of viable spermatogonia increased, reflecting an active supply of differentiated sper matogonia entering meiosis. In conclusion, apoptosis of Sertoli and pre-meiotic germ cells plays an important role in the acquisition of a suitable ratio of both cell types, and in providing intratubular environments for further progression of spermatogenesis, by controlling numbers of both cell types during the post-natal period.

[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.

Requirement of Notch 1 and its ligand jagged 2 expressions for spermatogenesis in rat and human testes.

It has already been demonstrated that the Notch signaling system is essential for gametogenesis in the adult germ line of Caenorhabditis elegans. However, the role of the Notch signaling system in mammalian spermatogenesis has not been well investigated. Recently, it has been revealed that this signaling system is expressed in the mammalian testis by showing coexpression of Jagged 2 and its receptor, Notch 1, is consistent with Notch 1 being a cognate receptor for Jagged 2 in the mammalian testis. Therefore, we investigated expressions of messenger RNAs of Notch 1 and Jagged 2 in the testicular tissues of developing Sprague-Dawley rats by reverse transcription-polymerase chain reaction and Northern blot analysis, expressions of their proteins in the testicular tissues of developing rats, fertile human controls and infertile human patients with maturation arrest by immunohistochemistry, and effects of antibodies to this system by culturing rat testicular tissues with these antibodies. Transcripts of Notch 1 and Jagged 2 in the rat testis were positive throughout the examined period; these intensities became higher at day 13 after birth, coincidence with the formation of spermatocytes, and peaked at day 19 after birth. Expressions of Notch 1 and Jagged 2 were recognized at first in the perinuclear regions of spermatocytes in the rat testis as a round structure at day 19 after birth and thereafter in further differentiated germ cells as meiosis proceeded. In the adult rat testis, positive staining was present as a round structure in spermatocytes, as a typical horseshoe-shaped structure in round spermatids, and as a covering structure spreading around the nucleus of elongated spermatids, but not in spermatozoa. Notch 1 was recognized in the vacuole of the Golgi complex of primary spermatocytes and the acrosome of elongated spermatids with electron microscopy. When rat testicular tissues were cultured with anti-Notch 1 or anti-Jagged 2 antibody, round and elongated spermatids decreased after 5 and 7 days of culture, respectively, and disappeared at around 9 and 12 days of culture, respectively, with shrinkage of the diameter of seminiferous tubules. Spermatocytes, however, increased after 11 days of culture. Expressions of both proteins have been detected in the testicular tissues of human fertile controls as in the rat testicular tissues. However, Notch 1 expression has not been detected in testicular tissues of 11 patients with maturation arrest, whereas Jagged 2 expression has been recognized in all of them. In conclusion, the results presented in this study offer the possibility that Notch 1/Jagged 2 signaling system plays an important role for male germ cells to differentiate or at least to survive in the rat testis and fails to express in the testis of spermatogenic maturation arrest patients.