Dual immunocytochemical staining of annexin A10 and p53 in low-grade and papillary urothelial carcinoma contributes to improvement of diagnostic accuracy in urine cytology.

BACKGROUND: Urothelial carcinoma (UC) is a common type of human cancer and, although urine cytology is a useful method for identifying high-grade UC (HGUC), its ability to diagnose low-grade UC (LGUC) is limited. The authors previously reported that annexin A10 (ANXA10) expression was strongly linked to both papillary and early stage LGUC and was inversely correlated with p53 expression in upper tract UC (UTUC) and bladder UC. However, it remains largely unknown whether ANXA10 is useful as a diagnostic marker for urine cytology. METHODS: In this study, the authors used 104 biopsy and 314 urine cytology samples to investigate the efficacy of ANXA10 and p53 expression by immunohistochemistry and immunocytochemistry. RESULTS: In immunohistochemistry analysis, expression levels of ANXA10 and p53 were either weak or absent in noncancerous tissues, whereas ANXA10 overexpression was observed patients with LGUC, and strong expression of p53 was identified in patients with HGUC. In immunocytochemistry analysis, sensitivity was not good for the detection of UC, especially UTUC, by cytology alone, but it was improved by combining cytology with ANXA10 and p53 to detect both bladder UC and UTUC. Receiver operating characteristic curve analysis also confirmed the diagnostic superiority of cytology combining ANXA10 and p53 for the detection of all UCs, including both HGUC and LGUC (area under the curve, 0.84). CONCLUSIONS: To the authors' knowledge, this is the first report that the combination of ANXA10 and p53 has potential application as a diagnostic immunomarker for improving the diagnostic accuracy of urine cytology.

A Case of Bilateral Synchronous Paratesticular Leiomyoma.

Bilateral synchronous paratesticular leiomyoma (BSPL) is a rare tumor that originates from smooth muscle cells in the paratesticular region. Four BSPL cases have been reported sporadically, starting with the 1991 report by Aus and Boiesen. Herein, we report the case of a 60-year-old male with a bilateral scrotal mass with a maximum size of 7.5 cm. Histological examination revealed oval to spindle-shaped tumor cells with a fascicular growth pattern. Immunohistochemically, the tumor cells were positive for α-smooth muscle actin. The pathological diagnosis was a leiomyoma. Based on the simultaneous bilateral nature of the disease, BSPL was diagnosed. In conclusion, we encountered a rare case of BSPL, and our report may contribute to the understanding of this disease.

[About Gonad Protection Based on Ovarian Position in Hip Plain Radiography].

PURPOSE: In gonad protection, it is difficult to identify the position from the body surface during shielding because the position and size of the ovary vary from individual to individual, and it is not possible to evaluate whether the protective equipment is correctly placed at the position of the ovary. Therefore, the position of the ovary with respect to the pelvis was clarified, and the effectiveness of gonad protection in the front and side of the hip joint was evaluated. METHODS: From the image of the pelvis taken with an MRI device, the inner and outer edges of the ovary, the upper and lower edges and the long and short axes of the pelvis, and the depth of the ovary were measured, and the position of the ovary was calculated based on the ratio of the ovary to the pelvis. A pelvic schema was created, and the position of the ovary was synthesized on the schema. In addition, the shielding rate was calculated when lead-containing rubber for the protection of the gonads was used. RESULTS: In front of the pelvis, the ovaries were present throughout the pelvic cavity. On the anterior surface, placing the shield on the caudal side up to the line connecting the centers of the left and right femoral heads had a shielding effect of about 88%. On the lateral side, shielding the pubic upper limbs from the ischial body could reduce the exposure of the unhealthy ovaries by 99%. However, when the gonad protection was placed at the height of the line connecting the anterior superior iliac spines, the shielding rate from the left and right ovarian distribution was about 13%, so the disadvantage of using the protective equipment was greater. CONCLUSION: For gonad protection, the presence or absence of use should be judged by using the shielding rate according to the shape of the protective equipment as an index.

Cytological and histological findings of upper tract mucinous urothelial carcinoma with clear cell component: A case report and review of literature.

Mucinous urothelial carcinoma (UC) is a rare variant and only 18 cases of mucinous UC have been reported. In this article, we report a case of mucinous UC focusing on both cytological and histological findings. A 92-year-old female was referred to our hospital because of gross hematuria. Clinical computed tomography scan showed 2.2-cm papillary lesion in the lower part of the left ureter. Urine cytology was performed, and cytopathological findings showed that there were a few atypical cells with pale to clear cytoplasm, and a low amount of mucin in the background was identified by periodic acid-schiff (PAS) and alcian blue (AB) staining. Laparoscopic radical nephrectomy of left renal pelvis and ureter was performed. The gross examination revealed that a white-gray, papillary-sessile tumor was found in the lower part of the left ureter. Histologically, conventional high grade UC cells were seen in some areas, and tumor cells in other areas showed abundant clear cytoplasm with extracellular and intracytoplasmic mucin. Immunohistochemical analysis revealed that tumor cells were positive for CK7, CK20, p63, GATA3, MUC1, MUC2, and MUC5AC and negative for MUC6 and CDX2. Histopathological diagnosis was mucinous UC with clear cell component, and the pathological stage was pT1N0M0. The patient has remained well and disease-free for 3 months after the operation. Familiarity and recognizing the characteristic pathological findings of mucinous UC are important because it represents a malignant neoplasm.

Application of stable isotope analysis for detecting filial cannibalism.

A novel type of filial cannibalism has been reported in pipefishes, in which the eggs are absorbed through the male's brood-pouch epithelium. The present study explored the applicability of stable isotope analysis for the detection of paternal brood cannibalism in the seaweed pipefish Syngnathus schlegeli. As expected, the δ15N values for liver, which conveys short-term dietary information about the recent reproductive season, were higher in males than in females. In contrast, the δ15N values for muscle, which reflects longer-term feeding habits that span both the reproductive and non-reproductive seasons, did not significantly differ between the sexes. This finding indicates that males occupy a higher trophic position than females only during the reproductive season, and it is probable that this difference is a result of paternal uptake of nutrients from embryos in the brood pouch.

Spectral tuning in sensory rhodopsin I from Salinibacter ruber.

Organisms utilize light as energy sources and as signals. Rhodopsins, which have seven transmembrane α-helices with retinal covalently linked to a conserved Lys residue, are found in various organisms as distant in evolution as bacteria, archaea, and eukarya. One of the most notable properties of rhodopsin molecules is the large variation in their absorption spectrum. Sensory rhodopsin I (SRI) and sensory rhodopsin II (SRII) function as photosensors and have similar properties (retinal composition, photocycle, structure, and function) except for their λ(max) (SRI, ∼560 nm; SRII, ∼500 nm). An expression system utilizing Escherichia coli and the high protein stability of a newly found SRI-like protein, SrSRI, enables studies of mutant proteins. To determine the residue contributing to the spectral shift from SRI to SRII, we constructed various SRI mutants, in which individual residues were substituted with the corresponding residues of SRII. Three such mutants of SrSRI showed a large spectral blue-shift (>14 nm) without a large alteration of their retinal composition. Two of them, A136Y and A200T, are newly discovered color tuning residues. In the triple mutant, the λ(max) was 525 nm. The inverse mutation of SRII (F134H/Y139A/T204A) generated a spectral-shifted SRII toward longer wavelengths, although the effect is smaller than in the case of SRI, which is probably due to the lack of anion binding in the SRII mutant. Thus, half of the spectral shift from SRI to SRII could be explained by only those three residues taking into account the effect of Cl(-) binding.

Tracing anthropogenic inputs to production in the Seto Inland Sea, Japan--a stable isotope approach.

The Seto Inland Sea (SIS) receives waste runoff from ∼24% of Japan's total population, yet it is also important in regional fisheries, recreation and commerce. During August 2006 we measured carbon and nitrogen stable isotopes of particulate organic matter (POM) and zooplankton across urban population gradients of the SIS. Results showed a consistent trend of increasing δ(15)N in POM and zooplankton from the western to eastern subsystems of the SIS, corresponding to increasing population load. Principal components analysis of environmental variables indicated high positive loadings of δ(15)N and δ(13)C with high chlorophyll-a and surface water temperatures, and negative loadings of low salinities related to inputs from large rivers and high urban development in the eastern SIS. Anthropogenic nitrogen was therefore readily integrated into the SIS food web from primary production to copepods, which are a critical food source for many commercially important fishes.

Living on the wedge: female control of paternity in a cooperatively polyandrous cichlid.

Theories suggest that, in cooperatively breeding species, female control over paternity and reproductive output may affect male reproductive skew and group stability. Female paternity control may come about through cryptic female choice or female reproductive behaviour, but experimental studies are scarce. Here, we show a new form of female paternity control in a cooperatively polyandrous cichlid fish (Julidochromis transcriptus), in which females prefer wedge-shaped nesting sites. Wedge-shaped sites allowed females to manipulate the siring success of the group member males by spawning the clutch at the spot where the large males were just able to enter and fertilize the outer part of the clutch. Small males fertilized the inner part of the clutch, protected from the large aggressive males, leading to low male reproductive skew. Small males provided more brood care than large males. Multiple paternity induced both males to provide brood care and reduced female brood care accordingly. This is, to our knowledge, the first documented case in a species with external fertilization showing female mating behaviour leading to multiple male paternity and increased male brood care as a result.

Niche differentiation depends on body size in a cichlid fish: a model system of a community structured according to size regularities.

1. Communities of different species are often structured according to niche differentiation associated with competitive interactions. We show that similar principles may apply on an ecological time-scale when individuals of a species having a wide size variation compete for resources, using the Lake Tanganyika cichlid Lobochilotes labiatus (5-30 cm). This species has a mouth especially adapted to suck up invertebrates from rock crevices. 2. Individuals defended feeding territories against similar-sized conspecifics, but not against different-sized ones. Thus, territories of similar-sized fish rarely overlapped, but up to a total of seven individuals (of seven size-ranks) had broadly overlapping territories with dissimilar-sized individuals. Comparison with expectation from the null model demonstrated clearly that observed size ratios between adjacent size rank were determined non-randomly regardless of sexual combinations. 3. Larger individuals took larger prey types of larger average size, but more importantly used wider rock crevices from which to suck food than smaller individuals. We calculated pairwise values of Schoener's index of diet overlap C(d) and the values of Levin's index of diet breadth B(d) (prey type and prey size) and the same for the width of the rock crevices used for foraging (C(r) and B(r)). C(d) remained high among all combinations of the seven ranks. In contrast, C(r) declined strongly in combinations of adjacent ranks (to 0.27), and was low or zero among further different size ranks. This shows that fish with overlapping territories divided the food resources largely through foraging site partitioning. Accordingly, B(d) did not depend on the size difference to the nearest two coinhabiting fish, whereas B(r) did. 4. We conclude that this L. labiatus community is structured non-randomly: body size-dependent effects on foraging site usage result in competition with, and territorial exclusion of, similar-sized individuals, but not of dissimilar-sized individuals that were accepted as coinhabitants. Accordingly, mean body size ratios (large/small) between two adjacent ranks were consistently approximately 1.28 [standard deviation (SD) = 0.07, n = 104], while approximately 1.34 from the null model (SD = 0.34, n = 10 400 simulations). We discuss our results as an example of Hutchinson's rule, applied originally to size ratios of different species.

Structure-based drug design of a highly potent CDK1,2,4,6 inhibitor with novel macrocyclic quinoxalin-2-one structure.

The design of a novel series of cyclin-dependent kinase (CDK) inhibitors containing a macrocyclic quinoxaline-2-one is reported. Structure-based drug design and optimization from the starting point of diarylurea 2, which we previously reported as a moderate CDK1,2,4,6 inhibitor [J. Biol.Chem.2001, 276, 27548], led to the discovery of potent CDK1,2,4,6 inhibitor that were suitable for iv administration for in vivo study.

Identification of potent 5-pyrimidinyl-2-aminothiazole CDK4, 6 inhibitors with significant selectivity over CDK1, 2, 5, 7, and 9.

5-Pyrimidinyl-2-aminothiazole 1 was identified as an inhibitor of cyclin-dependent kinases (CDKs) by a screening of the Merck sample repository. The introduction of a methyl group at the C-5 or C-6 position on the pyrimidine ring, directed toward the gate keeper residue of CDK4 (Phe93), led to significant enhancement of selectivity for CDK4 over other CDKs. Compound 3 exhibited more than 300-fold selectivity for CDK4 over CDK1, 2, 5, 7, and 9. Subsequent improvements in aqueous solubility afforded compound 4, which is available for further in vivo studies and this compound inhibited pRb phosphorylation and BrdU incorporation in tumor models.